Volume 14, Issue 1

Salix xerophila , S. maximowiczii , and S. koreensis are species of willow native to Korea that are important for bioenergy production. However, the native range of these species has narrowed in recent years due to the impact of climate change. Seeds of these Salix species lose viability within 4 weeks at ambient temperature, and within 4 months at -4°C. Preservation techniques are urgently needed to protect these valuable resources. The effects of seed water content (SWC; 3%, 6%, 9%, 12%, 18%, and 24%) and temperature (ambient, 4°C, -18°C, -80°C, and -196°C) on storage stability were investigated for up to 48, 52, or 60 months, depending on species. Optimal storage temperature and SWC varied between species. S. xerophila seed could be stored without deterioration for 60 months with 9% SWC at -80°C, but rapidly lost viability when stored at -18°C. In S. maximowiczii and S. koreensis , 100% and 90% of normal germination, respectively, was maintained with 18% SWC at -18°C or -80°C. Thus, for some Salix species, storage at -18 and -80°C may provide an economical alternative to cryopreservation or medium-term storage for the maintenance of seedbanks or breeding stocks.

The WRKY gene family is an ancient plant transcription factor (TF) family with a vital role in plant growth and development, especially in response to biotic and abiotic stresses. Although many researchers have studied WRKY TFs in numerous plant species, little is known of them in Tartary buckwheat ( Fagopyrum tataricum ). Based on the recently reported genome sequence of Tartary buckwheat, we identified 78 FtWRKY proteins that could be classified into three major groups. All 77 WRKY genes were distributed unevenly across all eight chromosomes. Exon–intron analysis and motif composition prediction revealed the complexity and diversity of FtWRKYs, indicating that WRKY TFs may be of significance in plant growth regulation and stress response. Two separate pairs of tandem duplication genes were found, but no segmental duplications were identified. Overall, most orthologous gene-pairs between Tartary and common buckwheat evolved under strong purifying selection. qRT-PCR was used to analyze differences in expression among four FtWRKYs (FtWRKY6, 74, 31, and 7) under salt, drought, cold, and heat treatments. The results revealed that all four proteins are related to abiotic stress responses, although they exhibited various expression patterns. In particular, the relative expression levels of FtWRKY6, 74, and 31 were significantly upregulated under salt stress, while the highest expression of FtWRKY7 was observed from heat treatment. This study provides comprehensive insights into the WRKY gene family in Tartary buckwheat, and can support the screening of additional candidate genes for further functional characterization of WRKYs under various stresses.

MicroRNAs (miRNAs) have been shown to play key roles in the regulation of plant growth and development by modifying the expression of their target genes. However, the influence of miRNAs on root formation and development in woody plants, such as Taxus chinensis , remains largely unknown. In the current study, we explored the phytohormone-response and nutrition-response miRNA expression profiles during T. chinensis rooting by quantitative real-time PCR (qPCR). We identified six phytohormone-response miRNAs, namely, miR164a, miR165, miR167a, miR171b, miR319, and miR391, and eight nutrition-response miRNAs, namely, miR169b, miR395a, miR399c, miR408, miR826, miR827, miR857, and miR2111a, that were differentially expressed at different rooting phases of T. chinensis . Using northern blot analysis of the putative target genes of these miRNAs, we detected the relative gene expression changes of the target genes. Taken together, our results suggest that miRNAs are involved in root formation of T. chinensis and that miRNAs may play important regulatory roles in primary root, crown root, and root hair formation by targeting phytohormone and/or nutrition response genes in T. chinensis . For the first time, these results expand our understanding of the molecular mechanisms of plant root formation and development in a conifer species.

Metabolomics allows the identification of biochemical markers that have important roles in plant resistance to pests and diseases by which breeders can select plants based on differences in these compounds. This study examines the range of compounds associated with plant defense against nematodes. Resistant tomato genotypes, GM2 and F1 (GM2 × Hawai 7996), and susceptible genotypes, Gondol Putih and Gondol Hijau, were used in this study. Peroxidase activity was measured colorimetrically using a spectrophotometer. 1 H-NMR (nuclear magnetic resonance) spectroscopy combined with orthogonal projections to latent structures discriminant analysis was used to analyze the metabolites involved in the tomato-nematode interactions. Identified signals were semi-quantitatively calculated by scaling the intensity of the 1 H-NMR to the signals of an internal standard (trimethyl silyl-3-propionic acid) at 0.00 ppm. Resistant plants showed a higher peroxidase activity than susceptible plants. Chemical compounds that differentiated between susceptible and resistant plants were glucose and caffeic acid. Resistant tomatoes were observed to have seven times higher level of glucose than susceptible plants. Glucose is the primary metabolite that acts in the signaling pathways in plant defense mechanisms. Caffeic acid is one of the phenolic compounds alleged to have a negative effect on the nematode.

Halophytes can survive and complete their life cycle in the presence of ≥200 mM NaCl. These remarkable plants have developed various strategies to tolerate salinity and thrive in high-salt environments. At the appropriate levels, salt has a beneficial effect on the vegetative growth of halophytes but inhibits the growth of non-halophytes. In recent years, many studies have focused on elucidating the salt-tolerance mechanisms of halophytes at the molecular, physiological, and individual level. In this review, we focus on the mechanisms, from the macroscopic to the molecular, underlying the successful growth of halophytes in saline environments to explain why salt has beneficial effects on halophytes but harmful effects on non-halophytes. These mechanisms include the specialized organs of halophytes (for example, ion compartmentalization in succulent leaves), their unique structures (salt glands and hydrophobic barriers in roots), and their salt-tolerance genes. We hope to shed light on the use of halophytes for engineering salt-tolerant crops, soil conservation, and the protection of freshwater resources in the near future.

Phosphate-solubilizing bacteria (PSB) can convert insoluble rhizosphere phosphorus into forms that are absorbable by plants and thus enhance the growth of plants. Safflower is a cash crop that is a source of vegetable oils, food coloring and flavoring agents. This study sought to isolate PSB in safflower rhizosphere soil and investigate their effects on seedling growth. The isolated PSB were identified as belonging to the genera Pseudomonas , Sinorhizobium , Staphylococcus , Acinetobacter and Enterobacter using 16S rRNA gene sequence analysis. Acinetobacter sp RC04. showed the best performance in phosphate solubilization, with the efficiency of the process being influenced by carbon source, nitrogen source, cultivation temperature and initial culture pH. Acinetobacter sp. RC04 and Sinorhizobium sp. RC02 showed the ability to improve safflower seed germination and, when co-inoculated, improved seedling growth. Hence, we suggest that Acinetobacter sp. RC04 and Sinorhizobium sp. RC02 could be developed for field application to promote safflower growth. The results from this study will help drive novel biofertilizer discovery and could be included in integrated nutrient management regimes for safflower and other important economic crops.

The perennial selenium (Se) hyperaccumulator Cardamine hupingshanensis (Brassicaceae) thrives in aquatic and subaquatic Se-rich environments along the Wuling Mountains, China. Using bright-field and epifluorescence microscopy, the present study determined the anatomical structures and histochemical features that allow this species to survive in Se-rich aquatic environments. The roots of C. hupingshanensis have an endodermis with Casparian walls, suberin lamellae, and lignified secondary cell walls; the cortex and hypodermal walls have phi (Φ) thickenings; and the mature taproots have a secondary structure with a periderm. The stems possess a lignified sclerenchymal ring and an endodermis, and the pith and cortex walls have polysaccharide-rich collenchyma. Air spaces are present in the intercellular spaces and aerenchyma in the cortex and pith of the roots and shoots. The dense fine roots with lignified Φ thickenings and polysaccharide-rich collenchyma in the shoots may allow C. hupingshanensis to hyperaccumulate Se. Overall, our study elucidated the anatomical features that permit C. hupingshanensis to thrive in Se-rich aquatic environments.

This study examines the effects of light emitting diodes (LEDs) on tissue culture proliferation of Acacia melanoxylon plantlets among five different clones (FM 1 , FM 2 , FM 4 , FM 5 , and FM 10 ). Shoot bud apex cuttings were transplanted onto Murashige and Skoog basal medium containing 0.1 mg L -1 6-benzyladenine and 0.5 mg L -1 naphthalene acetic acid and cultured in vitro for 40 days. Root growth was studied under different light intensities and photoperiods ex vitro . The bud proliferation coefficient was greatest under a light intensity of 45 μmol m -2 s -1 photosynthetic photon flux and photoperiod of 16 h light, but decreased as the light intensity increased. However, the greatest light intensity was beneficial for the growth of robust plantlets. Plantlets exposed to red and blue LED combinations grew tall and green, with a small number of roots. Plantlets also grew taller and some roots expanded under the longer photoperiod. Increased light intensity had positive effects on root number and rooting rate, and prolonged light greatly increased root number. Therefore, lower light intensity and a short photoperiod were beneficial for bud proliferation, while red/blue LED combinations, increased light intensity, and longer light illumination were beneficial for plantlet growth and root growth of Acacia melanoxylon .

The choice of particular trees for stripping by deer is puzzling because a preference is observed for trees of the highest social class. Trees ranked highest in the stand can produce more carbohydrates as a product of carbon assimilation. Among the many important nutrient and defense groups of metabolites in plants, high sugar content is postulated to be an attractant due to its impact on the sense of taste of different herbivores. Thus, we hypothesize that the carbohydrate content in the bast of the European beech is the most important factor by which deer make their choice. Our aim was to assess whether the sugar content in the bark and in the bast can be a factor which influences the intensity of bark stripping of particular trees in a beech thicket by red deer. A single episode connected with tree damaging pertained to 7% of the trees on the analyzed sampling plots. In total, 27% of the trees were damaged, including old damages. The mean size of damage to a given tree equaled 36% of the tree’s circumference and 0.06 m 2 of the trunk’s surface. Analysis of the sugar content in the bast and in the bark jointly indicates that the trees subjected to bark stripping possessed a distinctly higher sugar content than the trees which were not damaged. The probability of bark stripping grows together with an increase in the sugar content of the bark and of the bast.

The present study was conducted to evaluate the impact of monometallic and bimetallic nanoparticles (NPs) of copper (Cu) and silver (Ag) from Justicia spicigera on the photochemical efficiency and phenol pattern of Prosopis glandulosa . In this study, the existence of localized surface plasmon resonance absorption associated with the nano-sized nature of Ag, Cu and Cu/Ag particles was confirmed by the presence of a single peak around 487, 585, and 487/580 nm respectively. Zeta potential and electrophoretic mobility were found to be 0.2 mV and 0.02 μmcm/(Vs) for synthesized NPs indicating less stability and thus tendency to agglomerate, and broad distribution of particles. Cu-NPs and Cu/Ag-NPs demonstrate that the dispersed phase is stable and has a minimum particle size at zeta potentials above –30 mV. Changes in phenolic compounds, total chlorophyll, and photochemical efficiency in leaves exposed to Ag, Cu and Cu/Ag phyto-nanoparticles were evaluated up to 72 hours. The results revealed that Ag-NP and Cu-NP from J. spicigera at 100 mg/L showed significant reduction in chlorophyll, epidermal polyphenol content and photochemical efficiency of P. glandulosa . In contrast, the application of bimetallic Cu/Ag-NP from J. spicigera showed a positive impact on physiological parameters of P. glandulosa after 72 h of exposure.

A random transposition mutant library of B. gladioli UAPS07070 was analyzed for searching mutants with impaired microbial antagonism. Three derivates showed diminished antimicrobial activity against a sensitive strain. The mutated loci showed high similarity to the quorum sensing genes of the AHL-synthase and its regulator. Another mutant was affected in a gene coding for a LysrR-type transcriptional regulator. The production of toxoflavin, the most well known antimicrobial-molecule and a major virulence factor of plant-pathogenic B. glumae and B. gladioli was explored. The absence of a yellowish pigment related to toxoflavin and the undetectable transcription of toxA in the mutants indicated the participation of the QS system and of the LysR-type transcriptional regulator in the regulation of toxoflavin. Additionally, those genes were found to be related to the swarming phenotype. Lettuce inoculated with the AHL synthase and the lysR mutants showed less severe symptoms. We present evidence of the participation of both, the quorum sensing and for the first time, of a LysR-type transcriptional regulator in antibiosis and swarming phenotype in a strain of B. gladioli

Biocontrol is a cost-effective and environmentally friendly technique used in agricultural production. We isolated and screened a bacterial strain from the soils of a peach orchard with high yield. Using biochemical and physiological analysis as well as phylogenetic sequencing data, we identified a strain of Bacillus methylotrophicus , strain XJ-C. The results of our screening trials showed that XJ-C was able to suppress M. fructicola at an inhibition rate of 81.57%. Following the application of a 1×10 9 CFU/mL XJ-C strain suspension to the fruits, leaves, and shoots of peach trees infected with M. fructicola , the inhibition rate reached 64.31%, 97.34%, and 64.28%, respectively. Using OM and SEM, we observed that, under the inhibition of strain XJ-C, M. fructicola mycelium and spores were abnormally shaped. Under TEM, cell walls were transparent, organelles had disappeared, and the intracellular vacuole was deformed. Thus, XJ-C has the potential to be used in biocontrol.

Background Herbal medicine is widely used for the prevention and treatment of diseases worldwide including Palestine and may require long term usage. The level of some heavy metals and microbial contaminants in some of these medicinal plants consumed by Palestinians were studied in order to evaluate their quality. Methodology The level of metals including: Zinc, Cadmium, Lead and Copper were quantified by Atomic absorption spectroscopy (AAS). Moreover, the bacterial and fungal contaminations were tested for some of the selected plants in Palestine. The procedures of microbial and elemental testing of the plants followed USP. Results The result of the heavy metals testing showed that copper and cadmium were above the allowable limits in all the tested plants. Zinc metal was above the allowable limit in 78.9% of the tested samples. The microbiological results of the tested plants showed that 63.2% of the tested plants were contaminated by bacteria and 89.5% were contaminated by yeast. Conclusions Herbal medicine used in the Palestinian markets doesn’t meet the international requirement for heavy metal and microbiological limits. Therefore, urgent action has to be taken by the responsible authorities including the Ministry of health to implement importation and registration requirements and perform regular quality checks of sold and imported herbal medicines. Pharmacists as expert professionals must take an active role in selling and advising consumers about the quality and efficacy of the sold plants.

We investigated fungal communities colonising black cherry stumps. We tested the hypothesis that black cherry stumps of greater diameter should be characterised by more diverse fungal communities than stumps of smaller diameter. The material for analyses came from Podanin Forest District. DNA was extracted using a Plant Genomic DNA purification kit. The results were subjected to bioinformatic analysis and statistical analysis. The OTU sequences were compared using the BLAST algorithm with reference sequences from the UNITE database. In total, 8192 raw sequences were obtained from samples of black cherry stumps applying the Illumina sequencing technique. The results of the statistical analysis indicate a trend towards increased diversity in bigger black cherry stumps. The dominant share of fungi associated with wood decomposition indicates the progressing process of decomposition in stumps. Identification of the role and functions of the individual components of fungal communities colonising stumps may provide insight into the overall ecology of these organisms and provide a basis for improved plant protection, with a view to limiting the occurrence of black cherries in the future in undesirable locations outside their natural range.

The objective of the present study was to compare the effect of rhamnolipids on the microbial biomass content and the activity of dehydrogenases (DHA), acid phosphatase (ACP), alkaline phosphatase (ALP), and urease (URE) in soil contaminated with two types of coal tar creosote: type C and type GX-Plus. The experiment was carried out on samples of sandy clay loam under laboratory conditions. Coal tar creosote was added to soil samples at a dose of 0 and 10 g·kg −1 DM, along with rhamnolipids at a dose of 0, 10, 100, and 1000 mg·kg −1 DM. The humidity of the samples was brought to 60% maximum water holding capacity, and the samples were incubated at 20°C. Microbial and biochemical parameters were determined on days 1, 7, 21, and 63. The obtained results demonstrated that the addition of rhamnolipids did not result in any significant changes in the activity of the determined parameters in the uncontaminated soil. However, it was observed that the application of these biosurfactants, particularly at the dose of 1000 mg·kg −1 DM, largely decreased the effect of coal tar creosote on the determined parameters. Moreover, the microbial biomass and the activity of ALP and URE were found to be the best indicator of bioremediation of soil contaminated with coal tar creosote.

Foliar trichomes are specialized structures that have first contact with atmospheric particulate matter (PM), while their effects on PM accumulation are rarely investigated. In this study, epiphytic Tillandsia brachycaulos with foliar trichomes was investigated. Trichomes were removed artificially to investigate PM adsorption and resuspension. The results showed that the maximum PM (13.94±0.20 g•m -2 ) and natural PM content (10.65±1.10 g•m -2 ) on T. brachycaulos samples with foliar trichomes was significantly higher than those without foliar trichomes. After PM deposition on the leaf surface, more than 85% of PM was dispersed by wind in plants without trichomes, significantly higher than those with trichomes (30.2%). Therefore, the effects of foliar trichomes on PM were reflected not only on PM adsorption, but on PM resuspension. Foliar trichomes can decrease PM resuspension falling on the leaf surface and promote the accumulation of these particles.

Agar is a polysaccharide that primarily constitutes the cell wall of red algae. It is a good source of carbon and energy for many microbes. In the present study, an agarolytic bacterium, UQAD-3, was obtained from the waters of Al-Uqair, the Arabian Gulf, Al-Ahsaa, Saudi Arabia. UQAD-3 exhibited agarolytic activity when grown on agar as the sole source of carbon and energy. The strain was identified as Pseudoalteromonas ruthenica based on comparative analysis of the 16S rRNA, with 99.6% similarity. This finding was further confirmed by phylogenetic analyses based on 16S rRNA gene sequences, which highlighted that UQAD-3 was assembled within the Pseudoalteromonas clade and constituted a monophyletic subcluster with P. ruthenica, KMM 300 T . The strain was further characterised biochemically using the Biolog Gen III microtest system. UQAD-3 showed positive reactions to 16 (17%) of the 94 diverse traits assessed. Good growth was reported in 10% NaCl indicating its moderate halophilic nature. These observations indicate the agarolytic potential of the strain and opens new horizons for industrial applications in the future.

The aim of the study was to compare the influence of selected soil bioconditioners and traditional fertilisation P+K+Ca on the vitality and yield of Lolium perenne and on the microbiological state of soil. The study was conducted between 2008 and 2009 – it was based on a field experiment started in 2006, in Brody, at the Agricultural Experiment Station of the Poznań University of Life Sciences, Poland. The factorial experiment was conducted in a randomized block design, with three replications. Two experimental factors were used: - non-nitrogen fertilisation (Physio-Mescal G18, PRP-SOL, Effective Microorganisms, Effective Microorganisms + Ca, P+K+Ca); - nitrogen fertilisation (N0 and N200 kg ha -1 ). The following parameters were measured: the yield of dry matter of perennial ryegrass, the plants’ vitality (chlorophyll concentration), the count of selected groups of soil microorganisms (heterotrophic, oligotrophic, copiotrophic bacteria, actinobacteria, fungi), soil enzymatic activity (dehydrogenases, acid phosphatase), and soil pH. The experiment showed that the bioconditioners were not an alternative to traditional mineral fertilisation, especially to nitrogen fertilisation, as a basic yield factor, but they could be a very valuable supplement to this fertilisation, and help to maintain the right biological potential of soil and its fertility, especially in the places where no manure or other non-chemical fertilisers are used.

The aim of this study is to investigate changes in regulatory B cells (Bregs) and the expression of related cytokines such as interleukin-10 (IL-10) and transforming growth factor (TGF)-β in a mouse model of collagen-induced arthritis (CIA). A total 20 DBA/1 mice (6-8 weeks old) were randomly divided into control and CIA disease groups. For the CIA disease group, animals were injected intradermally with chicken collagen type II and complete Freund's adjuvant. The calculated arthritis index score of the CIA group was significantly higher than that in control group. Hematoxylin and eosin staining showed tumid synovial cells with irregular arrangement and obvious hyperplasia, with a high degree of inflammatory cell infiltration in CIA model group. Cytometric bead array technology and quantitative RT-PCR indicated that the levels of IL-10 and TGF-β in serum, and synovial cells were significantly increased in the CIA group. The proportion of Bregs in the spleen of the CIA group was significantly increased compared to the control group. In conclusion, our findings demonstrate that the number of Bregs and the expression of TGF-β and IL-10 are enhanced in mice with CIA.

Lactate (LA), an endogenous metabolite produced from pyruvate, can accumulate in skeletal muscle in certain conditions including major diseases, as well as during intensive exercise. Using differentiated C2C12 myotubes, we evaluated the early (1-h) and delayed (24-h) effects of LA (8 mM) on mechanisms involved in myogenesis or muscle atrophy, including 5'-adenosine monophosphate-activated protein kinase (AMPK)-mediated inhibition of protein synthesis through the mTOR/P70-S6K pathway, Akt-mediated inhibition of expression of the MAFbx atrophic factor by FOXO3a and expression of the myogenic transcription factors, MyoD, myogenin and myosin heavy chain. Although the early effects of LA overload were not significant on myogenic or atrophic mechanisms, LA treatment for 24 h significantly activated atrophic mechanisms but suppressed myogenesis in myotubes. In addition, LA overload for 24 h significantly suppressed the expression of Sirtuin 1 and peroxisome proliferator-activated receptor gamma coactivator-1 alpha. Consistent with LA-induced activation of atrophic mechanisms, the diameter of C2C12 myotubes treated with LA for 24 h, but not for 1 h, was significantly lower than in control myotubes. Thus, a sustained, but not a transient, LA overload could induce muscle atrophy through the regulation of AMPK- and Akt-mediated pathways, although further in vivo studies are needed to confirm this.

Objective The purpose of this study was to investigate the diagnostic performance of serum CK-MB, TNF-α and hs-CRP in children with viral myocarditis (VMC). Methods Fifty-six children with confirmed diagnosis of VMC were included in this study. Of the included 56 cases, 25 subjects were in acute and other 31 were in the recovery stage. A contemporaneous control group of 22 children were included for comparison. The serum concentration of CK-MB, TNF-α and hs-CRP were examined in both VMC and control groups. Results The serum concentration of CK-MB, TNF-α and hs-CRP were 31.77±9.48 (UL), 143.11±23.27 (ng/L) and 8.10±1.94(mg/L) for acute stage VMC; 12.72±4.99 (UL), 83.15±13.35 (ng/L) and 4.07±1.12 (mg/L) for recovery stage VMC; 8.11±3.20 (UL), 68.27±12.55 (ng/L) and 2.56±1.27 (mg/L) for control group respectively; The serum concentration of CK-MB, TNF-α and hs-CRP were significantly different between acute stage VMC, recovery stage VMC and control groups (p<0.05); Significant positive correlation between CK-MB and hs-CRP were found in acute stage VMC (r=0.54, p=0.01) and recovery stage VMC (r=0.37, p=0.04). Using serum CK-MB, TNF-α and hs-CRP as the reference, the differential diagnosis sensitivity for acute and recovery stage VMC were 87.10 (70.17-96.37)%, 87.10 (70.17-96.37)% and 77.42 (58.90-90.415)%; The specificity were 92.00 (73.97-99.02)%, 96.00 (79.65-99.90)% and 100.00 (86.28-100.00)% respectively. Conclusion Serum concentration of CK-MB, TNF-α and hs-CRP in children with VMC were significant increased especially in acute stage, which can be used as biomarkers for VMC diagnosis.

Objective To systematically investigate the correlation between the G>A polymorphism of the peroxisome proliferator-activated receptor γ coactivator 1α ( PPARGC1A or PGC-1alpha ) gene rs8192678 locus and the susceptibility to type-2 diabetes mellitus (T2DM). Methods The inclusion and exclusion criteria and retrieval strategies of original literatures were formulated. Then, subjects and free words “PPARGC1A”,”gene polymorphism”, and “T2DM” were retrieved from the PubMed, EMBASE, and Cochrane Library databases. Case-control studies on the G>A polymorphism of the PPARGC1A gene rs8192678 locus and susceptibility to T2DM were included for the meta-analysis. Results The number of cases in the T2DM group and control group was 5,607 and 7,596, respectively. The meta-analysis revealed that the PPARGC1A gene rs8192678 locus G>A polymorphism is associated with susceptibility to T2DM. There are differences in each group of genetic models, of which three groups of genetic models are highly significant. In the allele model, OR=1.249, 95% CI: 1.099-1.419, and P =0.001. In the dominant inheritance model, OR=1.364, 95% CI: 1.152-1.614, and P =0.000. In the additive inheritance model, OR=0.828, 95% CI: 0.726-0.945, and P =0.005. And one group is significant, in the recessive inheritance model, OR=1.187, 95% CI: 1.021-1.381, and P =0.026. Conclusion In Western Asian, South Asian, European and African populations, the A allele of the PPARGC1A gene rs8192678 locus may be one of the risk factors for T2DM.

Early diagnosis of hepatocellular carcinoma (HCC) greatly improves the survival and prognosisfor patients. In this study weevaluate the diagnostic promise of combining serum alpha-fetoprotein (AFP) expression with two potential biomarkers, serum glypican-3 (GPC3) and expression of the micro-RNA miR-122 for hepatitis C virus (HCV) related early-stage HCC. For this study serum samples from 47 patients with early-stage HCC, 54 chronic HCV (CH) carriers, 35 patients with liver cirrhosis (LC) and 54 health controls (HC) were collected. In addition to routine laboratory investigations, serum AFP, GPC3 and miR-122 were measured in all patients and healthy controls. Receiver operating characteristic (ROC) curves were used to present sensitivity and specificity for the biomarkers. The three markers were all significantly elevated in the serum samples from HCC patients. ROC curves showed the three markers had similar diagnostic capacities for distinguishing early-stage HCC from HCV-positive controls (LC + CH). In order to distinguish early-stage HCC from high-risk LC patients, the expression of miR-122 was superior to GPC3. Combination of the three markers as a panel showed a better diagnostic performance than any of the single markers (P <0.05). Overall, this study revealed that serum expression of GPC3 and miR-122 may be useful biomarkers to combine with serum AFP expression for the diagnosis of HCV related early-stage HCC.

Objective: The aim of our study was to assess the prognostic value of the ratio of neutrophil to lymphocyte (NLR) for patients with surgically treated endometrial cancer (EC). Methods: We retrospectively reviewed 510 EC patients who were surgically treated between January 2010 and December 2016. We used receiver-operating characteristic (ROC) curve analysis to identify an optimal cut-off for NLR in predicting overall survival (OS), cancer-specific survival (CSS) and disease-free survival (DFS). Nonparametric tests were used to determine the associations between NLR and clinicopathologic characteristics. The Kaplan–Meier method and Cox proportional-hazards regression were used for survival analysis. Results: With a cut-off of 2.47, the 510 patients were divided into low NLR (NLR <2.47) and high NLR (NLR ≥2.47). Elevated NLR was associated with advanced stage (P=0.039), increased histology grade (P=0.005) and lymph node metastasis (P=0.041). Multivariable analysis suggested that NLR was an independent prognostic marker for OS (hazard ratio [HR] 4.7; 95% confidence interval [CI], 1.5-14.1; P =0.006), CSS (HR 3.6; 95% CI, 1.1-11.5; P =0.028) and DFS (HR 2.3; 95% CI, 1.0-5.2; P =0.044). Conclusion: NLR may be an independent prognostic indicator for OS, CSS and DFS. It could help clinicians with preoperative risk stratification and treatment strategy tailoring.

Objective To investigate the HER2/c-erbB-2, epidermal growth factor receptor (EGFR) protein expression in gastric cancer and association with patients’ clinical pathology characteristics and prognosis. Methods HER2/c-erbB-2 and EGFR protein expression was examined by immunohistochemical assay in gastric cancer tissue and corresponding paired normal gastric tissue of 67 patients of gastric carcinoma. The HER2/c-erbB-2, EGFR protein positive expression rate in cancer tissue and normal gastric tissue were compared. The correlation between HER2/c-erbB-2, EGFR protein positive expression and patients’ clinical pathology characteristics and survival was evaluated. Results The positive expression rate of HER2/c-erbB-2 in the cancer and paired normal gastric tissues were 32.8% (22/67) and 4.5% (3/67), respectively with statistical difference (p<0.05). And the positive expression rate of EGFR in cancer and paired normal gastric tissues were 41.8% (28/67) and 5.9 (4/67), respectively, with statistical difference (p<0.05). HER2/c-erbB-2 positive expression in cancer tissue was significant correlated with the pathology grading (p<0.05), tumor invasion depth (p<0.05) and local regional lymph node metastasis (p<0.05); EGFR positive expression in cancer tissue was significant correlated with the tumor invasion depth (p<0.05) and local regional lymph node metastasis (p<0.05). The median survival time was 13.14 and 23.6 months respectively for HER2/c-erbB-2 positive and negative expression groups respectively with statistical difference ( HR=2.26, 9%CI:1.06-4.80, p<0.05). However, the median survival time was 15.47 and 22.87 months for EGFR positive and negative expression groups respectively, without statistical difference (HR=1.78, 9%CI:0.96-3.29, p>0.05). Conclusion Positive expression of HER2/c-erbB-2 and EGFR proteins in cancer tissue was significant higher than normal gastric tissue and have significant correlation with prognosis.

Objectives To investigate the neuropeptide Y (NPY) expression in the tissue of pelvic floor ligament and anterior vaginal wall in female patients with pelvic organ prolapse (POP) and stress urinary incontinence (SUI). Method Seventeen patients with POP, 6 with SUI, 13 with POP and SUI (POP&SUI), and 10 controls were included in this study from First Affiliated Hospital of JiNan University. Immunohistochemical assay was used to examine NPY expression in the tissue of round ligament, cardinal ligament of uterus, uterosacral ligament, and anterior vaginal wall. NPY expression were compared between POP, SUI, POP&SUI and controls. Results NPY was positive expressed in the round ligament, cardinal ligament of uterus, uterosacral ligament, and anterior vaginal subepithelial connective tissue. Compared with the control group, NPY expression in the round, cardinal, and uterosacral ligaments in patients with POP&SUI group was decreased with significant statistical difference (p<0.05). NPY expression in anterior vaginal wall was significantly decreased in POP, SUI, and POP&SUI groups compared to normal group (p<0.05). Compared to POP group, NPY expression in SUI and POP&SUI groups were significantly decreased (p<0.05), however the difference was not statistical different between SUI and POP&SUI groups (p>0.05). In POP and POP&SUI groups, the NPY expression in the cardinal ligament of uterus, uterosacral ligament, and anterior vaginal wall were negatively correlated with age (p<0.05), however, was not correlated with number of pregnancy, number of delivery, and BMI (p>0.05). Conclusions NPY expression was reduced in the round ligament, cardinal ligament of uterus, Uterosacral ligament, and vaginal anterior wall of the patients with POP and SUI. The decreased NPY expression may play an important role in the development of pelvic floordysfunction.

RAS protein activator like-1 (RASAL1) exists in numerous human tissues and has been commonly demonstrated to act as a tumor suppressor in several cancers. This study aimed to identify the functional characteristics of RASAL1 in ovarian adenocarcinoma and a potential mechanism of action. We analyzed RASAL1 gene expression in ovarian adenocarcinoma samples and normal samples gained from the GEO and Oncomine databases respectively. Then the relationship between RASAL1 expression and overall survival (OS) was assessed using the Kaplan-Meier method. Furthermore, the biological effect of RASAL1 in ovarian adenocarcinoma cell lines was assessed by Quantitative real time-PCR (qRT-PCR), Cell Counting Kit-8 (CCK-8), western blot, wound healing and transwell assay. The statistical analysis showed patients with higher RASAL1 expression correlated with worse OS. The in vitro assays suggested knockdown of RASAL1 could inhibit cell proliferation, cell invasion and migration of ovarian adenocarcinoma. Moreover, the key proteins in the mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK/ERK) signaling pathway were also decreased in ovarian adenocarcinoma cells with RASAL1 silencing. These findings provide promising evidence that RASAL1 may be not only a powerful biomarker but also an effective therapeutic target of ovarian adenocarcinoma.

Up until now, changes in biochemical and physiological parameters occurring a long time after stress are not yet elucidated. This is particularly the case for metals, some of which may considerably influence other branches of metabolism, such as bioenergetics and antioxidant defense. The aim of the current study was to investigate changes in levels of minerals (calcium and magnesium) and trace elements (copper and zinc) in erythrocytes and the liver of rats injected once or twice (modeling repeated stress) with adrenaline. The tissues were sampled 0.5 and 24 hours after the injection. A single injection of adrenaline in rats led to a dramatic increase in the levels of zinc (Zn), magnesium (Mg), and calcium (Ca) in their erythrocytes and liver, without a return to the control level (unstressed animals) after 24 hours. The levels of copper (Cu) increased 0.5 hour after a single adrenaline injection in erythrocytes and the liver, but returned to the control level after 24 hours. Double injection of rats with adrenaline led to an increase in the levels of Cu and Zn in their erythrocytes, and Mg in the liver, without a return to the control level after 24 hours. On the other hand, the double injection led to a drastic but transient increase in levels of Mg and Ca in erythrocytes, and Cu, Zn, and Ca in the liver. Thus, injection with adrenaline results in dramatic changes in levels of minerals and trace elements, which do not return to the control level after stress. Low doses of adrenaline lead to more stable changes in levels of essential metals.

Objective To investigate the effects of postconditioning ischemia on the expressions of the hippocampus neuron autophagy-related proteins LC3-II and Beclin-1 in rats following cerebral ischemia reperfusion. Methods A total of 128 male Sprague–Dawley rats were randomly divided into 4 groups: control, cerebral ischemia-reperfusion (IR), cerebral ischemia post-conditioning group (IP), and PI3K/Akt inhibitor (LY294002). The rat cerebral ischemia model was established by the improved Pulsinelli four vessel occlusion method. The durations across the platform and escape latent period were recorded using the water maze experiment. The changes in cell morphology and the number of surviving hippocampal neurons were detected by hematoxylin-eosin (HE) staining. The cells with Beclin-1 and LC3-II in the hippocampal region were detected by immunohistochemical staining and Western blotting. Results When compared with the IR at 48 and 72 h, the number of platform passes increased and the escape latency time was shortened. Consequently, the HE staining detected positive cells with LC3-II and Beclin-1 increased in number at each time point in immunohistochemistry and the expressions of the LC3-II and Beclin-1 proteins were improved in the IP (P < 0.05). Conclusions Cerebral ischemic post-conditioning promoted the expressions of autophagy-related proteins LC3-II and Beclin-1 while relieving the injuries caused by cerebral ischemia reperfusion.

To investigate how long non-coding RNAs DUXAP8 (LncRNA DUXAP8) influence the cell proliferation and invasion of non-small-cell lung cancer (NSCLC), we detected the expression levels of LncRNA DUXAP8 in lung cancer (LC) tissues, 4 LC-related cell lines (A549, SPC-A1, SK-MES-1 and NCI-H1299) and normal lung tissues via quantitative real-time PCR (qRT-PCR). Compared with normal lung tissue, LncRNA DUXAP8 was significantly up-regulated in NSCLC, especially in stage III / IV and diameter ≥ 3cm of lung cancer. Among 4 lung cancer cell lines, LncRNA DUXAP8 in A549 cells was the highest ( P <0.001). Construction of LncRNA DUXAP8 overexpression and LncRNA DUXAP8 knockout in A549 cell lines was further performed and subsequently injected into nude mice to build an in vivo tumor xenograft model. The results indicated that LncRNA DUXAP8 overexpression significantly promoted the A549 cells’ proliferation, enhanced invasion and induced tumor growth. Conversely, LncRNA DUXAP8 knockout significantly suppressed A549 cells’ proliferation, weakened invasion and inhibited tumor growth. Taken together, our results imply that LncRNA DUXAP8 is a potential regulatory molecular marker in non-small-cell lung cancer.

Objectives To explore the risk factors related to regional lymph node metastasis in cervical cancer and analyze the value of independent risk factors in predicting regional lymph node metastasis. Methods We retrospectively analyzed the clinical data of 699 patients who underwent surgery for stage IB1–IIA2 cervical cancer in Quanzhou First Hospital affiliated to Fujian Medical University from 2010 to 2016. The patients were divided into metastasis ( n = 92) and non-metastasis ( n = 607) groups based on the postoperative pathology of regional lymph node status. The relevant clinicopathological features of the metastasis and non-metastasis groups were compared through variance analysis and chi-square tests. Logistic regression was adopted to screen relevant independent risk factors of regional lymph node metastasis. Results In univariate analysis, International Federation of Gynecology and Obstetrics (FIGO) stages, serum squamous cell carcinoma antigen (SCC-Ag), histological type of squamous carcinoma and maximal tumor diameter were related factors for lymphatic metastasis in patients with cervical cancer. In multivariate analysis, SCC-Ag and histological type of squamous carcinoma were independent prognostic factors for lymphatic metastasis in patients with cervical cancer. Pre-treatment SCC-Ag serum levels, as a predictor of lymph node metastasis of cervical cancer, revealed a sensitivity of 62.07% (95% confidence interval (CI): 51.03–72.62%), specificity of 65.15% (59.07–70.89%), and area under the receiver operating characteristic (ROC) curve of 0.69 (95% CI: 0.61–0.76). Conclusions Cervical cancer patients whose pathological type is squamous carcinoma with high levels of SSC-Ag pre-operation are more likely to be diagnosed with regional lymph node metastasis. Standardized lymph node dissection should be implemented during operation.

Prurigo pigmentosa (PP) is an inflammatory dermatosis with unknown etiology. The clinical presentations of PP varies according to the stages of the disease. Rarely, the formation of numerous vesicles and bullae upon erythematous infiltrative plaques can be found during the entire clinical course. In the present case, a 29-year-old Chinese woman presented with a 6-year history of relapsing pruritic erythematous plaques and bulla on her neck, chest and back. Physical examination revealed multiple erythematous plaques and vesicles in combination with mottled pigmentation in a symmetrical distribution and reticular pattern on the nape of her neck, chest and back. Histological examination of the biopsy specimen collected from the bullous area of her chest indicated a lichenoid reaction with intraepidermal bulla. This inflammatory region is characterized by recruitment of lymphocytes, spongiosis, and a perivascular lymphohistiocytic infiltrate in the upper dermis. Direct immunofluorescence analysis for IgG, IgA, IgM and C3 was negative. The diagnosis of bullous prurigo pigmentosa was verified based on the clinical manifestation and pathological findings. Minocycline hydrochloride therapy (100mg/d) was initiated, and 3 weeks later the rash had completely disappeared, which resulted in pigmentation of the entire area. No recurrence was observed during the 4 years follow-up.

Objective This study aims to investigate the differences in the expression of hypoxia-inducible factor-1α (HIF-1α), N-myc downstream-regulated gene 2 (NDRG2) and epithelial mesenchymal transition (EMT)-related proteins in normal gastric tissues, gastric cancer tissues and lymph node metastasis. Methods Immunohistochemistry was used to detect the expression of HIF-1α, NDRG2, E-cadherin, Snail and Twist in normal gastric tissues, gastric cancer tissues and lymph node metastasis. Results In normal gastric tissues, HIF-1α was not expressed, NDRG2 was highly expressed. There was a significant between the expression of NDRG2 and Snail, as well as of NDRG2 and Twist. In gastric cancer tissues, there was no statistically difference between the expression of HIF-1α and E-cadherin, NDRG2 and E-cadherin. However, there was a significant difference in expression between the expression of HIF-1α and Snail, HIF-1α and Twist, NDRG2 and Snail, and NDRG2 and Twist. In lymph node metastasis tissues, we show that HIF-1α was highly expressed, while NDRG2 was not, and the difference between the expression of HIF-1α and E-cadherin, HIF-1α and Snail, HIF-1α and Twist was not significant. Conclusion HIF-1α may promote EMT, possibly by inhibiting the expression of NDRG2.

The aetiology of polycystic ovary syndrome (PCOS) remains uncertain and thus dedicated studies are still of much importance. Patients in this group are at high risk for metabolic syndrome, diabetes and ischemic heart disease. Our goal was to use a dietary intervention, facilitating the regression of the disease, through the observation of lipid and hormonal profiles, carbohydrate metabolic parameters and metabolomics of plasma fatty acids. There were 39 Caucasian women with PCOS aged 26.76 ±5.08 that qualified for this study. Fatty acid profiles were investigated using gas chromatography. The results of plasma fatty acids were compared with the initial results and the control group. A three-month caloric reduction diet with low glycemic index (GI) reduces the level of nervonic acid and is a great alternative in PCOS therapy. The introduction of rapeseed oil and olive oil to the lowered GI reduction diet caused the increase in the ratio of average length chain fatty acids (C10:0, C14:0) and the enhancement of synthesis pathways for pentadecanoic acid (C15:0) and gamma-linolenic acid (GLA, C18:3n-6), but did not inhibit the synthesis of the derivatives of arachidic acid (C20:0). Additionally, a decrease in the level of nervonic acid (C24:1) was observed. Biochemical analysis of blood showed the improvement of plasma lipid fractions, but a significant reduction of androgen levels was not observed. A reduction diet with lowered GI lead to many positive effects in the improvement of the biochemical parameters of women with PCOS. It should be continued for a prolonged period of time, until the synthesis pathways for inflammatory factors are silenced.

Objective Ventral anterior homeobox 2 (VAX2) gene is a key regulating factor for the development of the ventral region of the eye, and has recently attracted much attention from the cancer treatment field. Our study aimed to explore the effect of VAX2 on papillary thyroid carcinoma (PTC). Methods We determined the expression levels of VAX2 in PTC based on The Cancer Genome Atlas (TCGA) database. We then assessed the prognosis of patients with PTC, and analyzed the association between VAX2 expression and clinicopathological characteristics. Subsequently, we measured the biological functions of VAX2 in PTC using qRT-PCR, cell counting kit-8 (CCK-8) assay, colony formation assay, wound healing assay, transwell assays and western blot. Results VAX2 was up-regulated in PTC tissues when compared with normal thyroid tissues, and high expression level of VAX2 was positively correlated with poor prognosis. Furthermore, knockdown of VAX2 significantly inhibited the proliferation, migration and invasion of PTC cells. Importantly, through western blot analysis, we found that the expression of phosphorylated-(p) ERK and p-MEK in ERK signaling pathway showed a significant decrease after knockdown of VAX2. Conclusion These findings suggest that VAX2 may be involved in the malignant progression of PTC, and hold significant potential as a therapeutic target for PTC.

Objective The aim of this study was to investigate the clinical effects of insulin resistance (IR) in the development of mild cognitive impairment (MCI) in elderly adults with Type 2 diabetes mellitus (T2DM). Methods Seventy-eight patients with T2DM were recruited and divided into MCI group (<26, n=48) and normal group (≥26, n=30) according to the Montreal Cognitive Assessment (MoCA) score. The fasting plasma glucose (FPG), HbA1c, and fasting plasma C-peptide (FPC) were examined and compared between the two groups. The Pancreatic islets function (HOMA-islet) and Insulin Resistance Index (HOMA-IR) were also calculated for the two groups. Using the HOMA-IR and HOMA-islet as the reference, the predicted values for MCI in T2DM patients were calculated by sensitivity, specificity and area under the receiver operating characteristic (ROC) curve. Results The MoCA scores were statistically different between the MCI and control groups (23.79±1.15 vs 28.50±1.01, p<0.05). The serum FPG and FPC were 10.38±2.36 mmol/L and 0.79±0.34 ng/mL in the MCI group which were significant different from those of the control group (8.96±2.55 mmol/L and 1.04±0.38 ng/mL; p<0.05). The HOMA-IR and HOMA-islet were 10.08±2.64 and 94.67±29.12 for the MCI group and 8.16±2.46 and 130.30±38.43 for the control group; both were statistically different (p<0.05). The serum HbA1c was 11.02±2.59% and 9.37±2.00% for the MCI and control groups (significantly different with p<0.5). A significant positive correlation was found between MoCA score and HOMA-islet (rpearson=0.44; p<0.001). A significant negative correlation existed between MoCA score and serum HbA1c (r=-0.25; p=0.03). The areas under the ROC curve were 0.70 (0.57~0.82), 0.69 (0.57~0.81), 0.69 (0.57~0.80), 0.72 (0.60~0.84), 0.72 (0.60~0.84) and 0.76 (0.65~0.88) respectively for FPG, FPC, HbA1c, HOMA-IR and HOMA-islet. Conclusion Insulin resistance is a risk factor for mild cognitive impairment and can be a biomarker for prediction of MCI in patients with T2DM.

Objective Mitochondrial homeostasis is vital for the progression of lung cancer. Nurr1 has been identified as a novel mediator of mitochondrial homeostasis in several types of cancers. The aim of our study was to investigate whether Nurr1 modulates the viability of A549 lung cancer cells by inducing mitochondrial dysfunction, with a focus on the p53-Drp1 signaling pathway. Methods western blotting, ELISA and immunofluorescence assay was used to verify the alterations of cell death. siRNA was used to determine the role of p53-Drp1 pathway in lung cancer death. Results Nurr1 was downregulated in A549 lung cancer cells compared to normal pulmonary epithelial cells. Interestingly, overexpression of Nurr1 reduced the viability of A549 lung cancer cells by activating apoptosis and mitochondrial stress. At the molecular level, we provide data to support the regulatory effects of Nurr1 on the p53-Drp1 signaling pathway. Blockade of the p53-Drp1 signaling pathway abolished the proapoptotic action of Nurr1 on A549 cells and sustained mitochondrial homeostasis. Conclusion Taken together, our results depict the tumor-suppressive role played by Nurr1 in A549 lung cancer in vitro and show that the anticancer effects of Nurr1 are executed via triggering of mitochondrial dysfunction and activation of the p53-Drp1 signaling pathway.

Objective The incidence of papillary thyroid carcinoma (PTC) is increasing, and there are no reliable serum biomarkers for the diagnosis and prognosis of PTC. This study aimed to assess whether serum matrix metalloproteinase-9 (MMP-9) could serve as an auxiliary diagnostic/prognostic marker for PTC after total and partial thyroidectomy. Material and Methods Postoperative serum MMP-9 concentrations were measured in 182 male patients with PTC, 86 male patients with benign thyroid nodule (BTN), and 62 male healthy controls (HCs). Multivariate logistic regression and Cox regression were applied to evaluate the correlation between variables. The performance of serum MMP-9 in diagnosing PTC and predicting structural persistent/recurrent disease (SPRD) during 48 months of follow-up after initial surgery was evaluated by receiving operating characteristic curve analysis. Results The median serum MMP-9 concentration in the PTC group (79.45 ng/ml) was significantly higher than those in the BTN group (47.35 ng/ml) and HC group (47.71 ng/ml). The area under the curve (AUC) for predicting PTC from BTN was 0.852 at a cut-off value of 60.59 ng/ml. Serum MMP-9 was negatively correlated with disease-free survival (OR 1.026, P=0.001). Serum MMP-9 exhibited good performance in predicting SPRD at a cutoff value of 99.25 ng/ml with an AUC of 0.818. Advanced TNM stage (OR 31.371, P=0.019) and serum MMP-9 ≥99.25 ng/ml (OR 4.103, P=0.022) were independent risk factors for SPRD. Conclusions Serum MMP-9 potentially represents a good predictive biomarker for PTC diagnosis and prognosis after thyroidectomy in Chinese male patients for whom radio-imaging indicates suspected PTC.

Changes in cellular metabolism, development of oxidative-nitrative stress and intensification of glycation and lipid peroxidation (LPO), are significant processes that occur during diabetes mellitus (DM)-associated chronic hyperglycemia. These processes contribute to deviations in the structural organization and functional activity of leukocytes. The development of oxidative-nitrative stress in peripheral blood cells during DM can be prevented by agmatine, an endogenous metabolite of L-arginine, which is a nitric oxide synthase (NOS) inhibitor, and possesses hypoglycemic properties. The administration of agmatine to animals with DM lead to the inhibition of both constitutive and inducible NOS in leukocytes, which in turn decreased total nitrite/nitrate (NOx) levels. Additionally, we observed corresponding increases in reduced glutathione content and activity of antioxidant enzymes (SOD, CAT, GPx, GR), along with decreased levels of the thiobarbituric acid reactive substance, advanced oxidation protein products (AOPPs) and advanced glycosylation end-products (AGEs) as compared to the non-treated diabetic group. Our results indicate that treatment of diabetic animals with agmatine restores redox homeostasis and a balances antioxidant defence system enzymes in leukocytes. This corrective effect on the functional capacity of leukocytes is exerted by preventing oxidative-nitrative stress in animals with DM.

Platelet-rich plasma (PRP) has been shown to be a beneficial growth factor for bone tissue healing and is used in implantology. The aim of this study was to investigate the effects of PRP on bone defects in rabbits. Twenty rabbits were used to establish the implant bone defect model in this study. An intrabony defect (5mm × 5mm × 3mm) was created in alveolar bone in the lower jar of each rabbit. The wound was treated with PRP. The expression of platelet-derived growth factor BB (PDGFBB) was assessed by enzyme-linked immunosorbent assay (ELISA). Focal adhesion kinase (FAK) and related phosphatidylinositol 3-kinase (PI3K)/AKT (protein kinase B) levels were measured by Western blot. The results show that PRP could significantly improve the bone healing process when compared with control, and 10% PRP could markedly increase fibroblast proliferation 48-h post treatment. PDGFBB was higher in the PRP group than that in the control group. PRP treatment also could elevate the phosphorylation of FAK and PI3K/AKT, although the inhibitor of PDGFR could reverse this trend. These results suggest that PRP treatment improves the bone healing process through the FAK/PI3K/AKT pathway.

Objective The aim of this study was to investigate the diagnostic efficacy of thrombelastography (TEG) in patients with preeclampsia. Methods One hundred and seventeen pregnant women were recruited from Department of Obstetrics and Gynecology of 1 st affiliated Hospital of Fujian Medical University. Of the 117 patients, 59 were normal late gestation (control group), 32 were mild preeclampsia and other 26 cases were severe preeclampsia. All the patients were received thrombelastography (including: K time, Reaction time, Clot angel, MA value, CI value) and blood coagulation examination (including: PT, APTT, Fib, TT, D-dimer and AT-III). Results The R time, K time, Coagulation Index value and Clot Angle in preeclampsia group were significant different between control and preeclampsia groups with statistical difference (p<0.05). Moreover, the R and K time value in severe preeclampsia group were significant higher than those of control groups (p<0.05); however, the Coagulation Index value and Clot Angle in severe preeclampsia group were significant higher than those of mild preeclampsia group with statistical difference (p<0.05). Coagulation Index had the highest diagnostic sensitivity [87.93 (76.70-95.01) %] and specificity [83.83 (79.17-96.18)%] compared to other parameters with the AUC of 0.94 (0.90-0.98). The K time and the Coagulation Index had the highest diagnostic sensitivity (96.15%) and specificity (0.75%) respectively with the AUC of 0.68 and 0.75 respectively in differential diagnosis of severe preeclampsia from mild preeclampsia. However, there were no statistical difference in the aspects of platelet count and parameters relevant to coagulation test for the control, mild and sever preeclampsia groups(p>0.05). Conclusion TEG provides more accurate information in monitoring the blood coagulation of preeclampsia patients and can be used as a reliable marker for assessing the severity of preeclampsia.

The cerebrospinal fluid content was examined for concentrations of S100 protein and neuron-specific enolase (NSE) in two diseases, Kawasaki disease (KD) with aseptic meningitis (1-3 months) and purulent meningitis (PM), to determine whether or not these measuremets could be used in early diagnosis. The content of cerebrospinal fluid S100 protein of KD with aseptic meningitis and PM were significantly higher than those in the control group. There was also a difference between KD and purulent meningitis (PM). The concentration of NSE was highest in the encephalitis group, which was statistically different from control group. However, there was no difference between the KD and control groups. The levels of S100 protein and NSE of KD with aseptic meningitis were lower than those in PM, indicating that the extent of neuronal damage is significantly lower than of the enchephalitis group. The area under the curve (AUCs) of the receiver operating characteristic (ROC) curve for S100 and NSE were both 0.972. The S100 threshold was 0.4315, the sensitivity was 92.1%, and the specificity was 100%, while the NSE threshold was 9.325, sensitivity 92.1%, and specificity 90%. The combined detection of NSE and S100 levels in the cerebrospinal fluid can be used for the differential diagnosis of KD with aseptic meningitis and purulent meningitis.

It has been reported that endocannabinoid receptor type 2 (CB2) agonist JWH133 inhibits the growth of C6 glioma cells, but the underlying mechanism has not yet been fully elucidated. We showed that JWH133 inhibited C6 cells growth, reduced cAMP production and inhibited PKA activity through CB2 receptor. Decrease of PKA activity stimulated CaMKKβ, and subsequently elevated phosphorylation of AMPKα at threonine 172 site. The activation of AMPKα induced changes of downstream proteins, including increase of P53 phosphorylation and P21 production, as well as decrease of mTOR phosphorylation, that eventually inhibited C6 cells growth.

This study examined whether or not various mouthwashes have significant effects on the viability or morphology of mouse osteoblast-like cells. Mouse calvarial preosteoblast cells were cultured and prepared, then treated with a 0.12% chlorhexidine digluconate solution containing essential oils with or without alcohol, and a cetylpyridinium chloride solution, and sodium fluoride, respectively. Each well was treated with one of six mouthwashes for either 30 sec, 1.5 min, or 4.5 min. The viability of the treated cells was quantitatively analyzed by a Cell Counting Kit-8. The viability of osteogenic progenitor cells decreased significantly irrespectively of the types of mouthwashes. The changes of cell morphology were seen in all groups of mouthwashes; however, they were more noticeable on the chlorhexidine digluconate-treated group. A progressive increase in treatment time over 30 sec did not seem to deteriorate cellular viability. There was no significant difference in viability or morphological change between different formulations of the same brand. Although various mouthwashes without alcohol as an ingredient are available, nonalcoholic mouthwashes were not likely to be less harmful to the cells. Collectively, commercially available mouthwashes could inhibit cell viability and alter the morphology of osteoblastic precursor cells irrespectively of brands, treatment time, or alcohol content.

Background Both glucose-regulated protein 78 kDa (GRP78) and glucose-regulated protein 94 kDa (GRP94) are important molecular chaperones that play critical roles in maintaining tumor survival and progression. This study investigated the effects in prostate cancer cells following the downregulation of GRP78 and GRP94. Methods RNA interference was used to downregulate GRP78 and GRP94 expression in the prostate cancer cell line, PC-3. The effects on apoptosis and cell migration was examined along with expression of these related proteins. Results Small interfering RNAs targeting GRP78 and GRP94 successfully down-regulated their expression. This resulted in the induction of apoptosis and inhibition of cell migration. Preliminary mechanistic studies indicated that caspase-9 (cleaved) and Bax expression levels were upregulated while Bcl-2 and vimentin expression levels were downregulated. Conclusion Co-downregulation of GRP78 and GRP94 expression induces apoptosis and inhibits migration in prostate cancer cells.

Objective The objective of this research is to investigate the expression and function of SKA3 in lung adenocarcinoma. Methods The mRNA expression level of SKA3 in lung adenocarcinoma and its association with clinic-pathological factors were analyzed using data obtained from the TCGA database. Small interfering RNA (siRNA) for SKA3 (si-SKA3) was used to down-regulate SKA3 in A549 cells. pcDNA3.1- SKA3 was used to overexpress SKA3 in A549 cells. The proliferation ability of A549 cells was determined via MTT assay and colony formation assay. A wound healing assay was performed to examine the migration ability of A549 cells. The protein expression of p-MEK, MEK, p-ERK and ERK were determined by western blot. Results We found that SKA3 is up-regulated in lung adenocarcinoma compared to the normal lung tissues. Kaplan-Meier analysis showed that high SKA3 expression is markedly associated with poor prognosis in lung adenocarcinoma patients. SKA3 expression is significantly correlated with age, gender, pathologic-stage, pathologic-N and pathologic-M. Moreover, depleting SKA3 obviously inhibited A549 cell proliferation and migration in vitro , while overexpression of SKA3 notably increased A549 cell proliferation and migration. Western blot analysis showed that the protein expression ratio of p-MEK/MEK and p-ERK/ERK decreased noticeably after depleting SKA3. Conclusion SKA3 expression was enhanced and associated with poor prognosis in lung adenocarcinoma patients, and it might play a facilitating role in cell growth and motility by regulating the MAPK signaling pathway.

To explore protective effects and related mechanisms of microRNA-182 (miR-182) on oxidative stress in rat hippocampal neurons (RHiN), RHiN cells. As the results, the survival rate and superoxide dismutase levels in H 2 O 2 group were significantly lower than H 2 O 2 +miR-182 group (all P<0.05). The malondialdehyde levels and apoptosis rate in H 2 O 2 +miR-182 group were significantly lower than H2O2 group (all P<0.05). The mRNA levels and expression levels of mTOR and PI3K in H 2 O 2 +miR-182 group were higher than those in H 2 O 2 group (both P<0.05). The experiment of cerebral ischemic oxidative stress model rats showed that the survival rate, apoptosis rate, malondialdehyde and superoxide dismutase levels in miR-182 group were better than model control group. The positive staining intensity of phosphoinositide 3-kinase (mTOR) and phosphoinositide 3-kinase (PI3K) in model control group were significantly lower than miR-182 group (all P<0.05). Increased levels of miR-182 can reduce the damage of H2O2 treatments in RHiN cells. Oxidative stress is decreased in the neuronal cells possibly by activation of the PI3K-AKT-mTOR pathway.

Venereal syphilis is a sexually transmitted disease caused by Treponema pallidum – Gram-negative, slowly growing bacteria. The spread of the disease in the Old World was due to increased birth rate, urban population growth, migration and lack of knowledge concerning the epidemiology. In the past, the treatment was mainly symptomatic and included application of mercury compounds. The goal of the study was to present the case of advanced venereal syphilis found in early modern (16th–18thc) graveyard localized in Wroclaw, Poland. The object of the study is a cranium of a male whose age at death has been estimated to be over 55. In order to observe the morphological and paleopathological characteristics of the examined material, anthropometrics, computed tomography, spectrometry and microscopic methods were incorporated. Microscopic analysis revealed the presence of the extensive inflammatory lesions. Analyses indicate tertiary stage of venereal syphilis as the most probable cause of the observed lesions. Concentration of arsenic (16.17±0.58 μg/g) in examined bone samples was about hundred times bigger than average arsenic concentration in bones reported in other studies. Advanced stage of observed lesions along with high arsenic level may suggest long-lasting palliative care and usage of arsenic compound in therapeutic treatment of this chronic disease.

LBHD1 (C11ORF48) is one of the ten potential tumor antigens identified by immunoscreening the urinary bladder cancer cDNA library in our previous study. We suspect that its expression is associated with human bladder cancer. However, the exact correlation remains unclear. To address the potential functional relationship between LBHD1 and bladder cancer, we examined the LBHD1 expression at the mRNA and protein level in 5 different bladder cancer cell lines: J82, T24, 253J, 5637, and BLZ-211. LBHD1 high and low expressing cells were used to investigate the migration, invasion, and proliferation of bladder cancer cells following transfection of LBHD1 with siRNA and plasmids, respectively. Our experiment showed that the degree of gene expression was positively related to the migration and invasion of the cancer cells while it had little effect on cell proliferation. Knocking down LBHD1 expression with LBHD1 siRNA significantly attenuated cell migration and invasion in cultured bladder cancer cells, and overexpressing LBHD1 with LBHD1 cDNA plasmids exacerbated cell migration and invasion. Nevertheless, a difference in cell proliferation after transfection of LBHD1 siRNA and LBHD1 cDNA plasmids was not found. Our findings suggest that LBHD1 might play a role in cell migration and invasion.

Targeting 6-phosphogluconate dehydrogenase (6PGD) can inhibit cancer cell proliferation and tumor growth. However, the relationship between 6PGD and cisplatin resistance still needs further study. Cisplatin-sensitive and cisplatin-resistant ovarian cancer OV2008 and C13* lines and lung cancer A549 and A549DDP lines were treated with different concentrations of cisplatin and cell viability was evaluated. We also compared the growth rates and the cell cycle distributions between cisplatin-sensitive and cisplatin-resistant cells. The expression level of 6PGD was detected by immunoblotting. The Chou-Talalay method was used to evaluate the effect of a combination treatment using cisplatin and the small molecule inhibitor 1-Hydroxy-8-methoxy-anthraquinon (S3) that targets 6PGD. The cisplatin-resistant ovarian and lung cancer cell lines grew faster than the cisplatin- sensitive cell lines, with more cells in S and G2 phases in cisplatin-resistant cell lines. The expression level of 6PGD in cisplatin-resistant cell lines was significantly increased compared with cisplatin-sensitive cell lines. Furthermore, treatment of cells with the S3 small molecule inhibitor of 6PGD together with cisplatin could overcome cisplatin resistance. The expression level of 6PGD in cisplatin-resistant cells lines was significantly upregulated, and the resistance to cisplatin of drug-resistant cells lines could be overcome when treated with the small molecule inhibitor S3 that specifically targets 6PGD.

Andrographolide (ANDR), isolated from Andrographis paniculata , is a medicinal compound effective against infections, inflammatory disorders, and various cancers. In the present study, the effects of ANDR on NFkB (nuclear factor kappa-light-chain-enhancer of activated B cells) activation, caspase-8-mediated apoptosis and pyroptosis, and extra cellular matrix (ECM) degradation were analyzed in A431, MDA-MB231, and SKOV-3 cell lines. Results showed that ANDR inhibited the growth and proliferation of cancer cell lines by inhibiting NFkB signaling. A significant decrease in phospho-p65 level was observed upon increasing ANDR concentration in epidermoid carcinoma and breast cancer cell lines, A431 and MDA-MB231, respectively. Accordingly, upon ANDR treatment, the expression of caspase-8 was increased, whereas no significant induction of caspase-1 expression was observed. Moreover, we observed a significant increase in the expression of tissue inhibitor of metallopeptidase-1 (TIMP1) upon increasing ANDR concentration. Such induction of TIMP1 inhibited the activity of matrix metallopeptidase-7 (MMP-7), thus preventing the degradation of ECM. Therefore, as ANDR shows cytotoxicity towards cancer cells via the NFkB signal transduction pathway without inducing pyroptosis and blocks breast and ovarian cancer invasion by inhibiting MMP-7 expression through TIMP1 up-regulation, it has the potential to be developed as a drug targeting ovarian and breast cancers.

The sialylation of the glycoproteins in skeletal muscle tissue is not well investigated, even though the essential role of the sialic acids for the proper muscular function has been proven by many researchers. The invasion of the parasitic nematode Trichinella spiralis in the muscles with subsequent formation of Nurse cell-parasite complex initiates increased accumulation of sialylated glycoproteins within the affected area of the muscle fiber. The aim of this study is to describe some details of the α-2,6-sialylation in invaded muscle cells. Asynchronous invasion with infectious T. spiralis larvae was experimentally induced in mice. The areas of the occupied sarcoplasm were reactive towards α-2,6-sialic acid specific Sambucus nigra agglutinin during the whole process of transformation to a Nurse cell.The cytoplasm of the developing Nurse cell reacted with Helix pomatia agglutinin, Arachis hypogea agglutinin and Vicia villosa lectin-B4 after neuraminidase pretreatment.Up-regulation of the enzyme ST6GalNAc1 and down-regulation of the enzyme ST6GalNAc3 were detected throughout the course of this study. The results from our study assumed accumulation of sialyl-Tn-Ag, 6`-sialyl lactosamine, SiA-α-2,6-Gal-β-1,3-GalNAc-α-Ser/Thr and Gal-β-1,3-GalNAc(SiA-α-2,6-)-α-1-Ser/Thr oligosaccharide structures into the occupied sarcoplasm. Further investigations in this domain will develop the understanding about the amazing adaptive capabilities of skeletal muscle tissue.

Aim This study investigates the effect of astragalus polysaccharides (APS) in protecting against thapsigargin-induced endoplasmic reticulum (ER) stress in HT29 cells by suppressing the PERK-eIF2a signaling pathway. Methods HT29 cells were induced by thapsigargin for 12 hours, then treated with APS for 24 hours, and the gene expressions of GRP78, CHOP and eIF2a were quantified by reverse transcription quantitative polymerase chain reaction (RT-qPCR). The expression of GRP78, CHOP, PERK, p -PERK, eIF2a, and p -eIF2a were detected by Western blot. Results The ER stress caused by thapsigargin strongly up-regulated the expression of GRP78 and CHOP in HT29 cells, which activated the PERK-eIF2a pathway. There was an increase in PERK phosphorylation, and induction of eIF2a in HT29 cells. Thapsigargin caused significant ER expansion in HT29 cells due to the 12-hour ER stress. Importantly, Astragalus polysaccharide significantly inhibited the phosphorylation of PERK and eIF2a, which reduced the mRNA levels of GRP78, CHOP, PERK and eIF2a, and inhibited the ER expansion in HT29 cells after 24 hours of treatment. Conclusion The results indicate that APS reduces the expression of GRP78 and CHOP in HT29 cells, at least in part, by preventing the activation of the PERK-eIF2a signaling pathway.

Hyperglycemia-induced toxicity of neurons contributes to the pathogenesis and progression of diabetic neuropathy (DNP). High concentration glucose triggered reactive oxygen species (ROS) overproduction and induced cell apoptosis of neurons from dorsal root ganglion (DRG) in vitro. Currently, there is no effective therapeutic method to retard this devastating complication or neurotoxicity induced by high glucose. Insulin-like growth factor-1 (IGF-1) has multi-neurotrophic actions which need to be explored regarding its actions and mechanisms on relieving high glucose induced neurotoxicity. Herein, high concentration glucose was exposed to the DRG neurons in vitro. The effects of IGF-1 on relieving high glucose-induced neurotoxicity were evaluated. We illustrated that IGF-1 enhanced regeneration of neurites sent from DRG neuronal cell bodies and increased neuronal viability which inhibited by high glucose challenge. IGF-1 alleviated neuronal apoptosis caused by high glucose exposure. IGF-1 also suppressed the intracellular ROS overproduction and ATF3 expression upregulation which was induced by high glucose insult. The anti-neurotoxic effects of IGF-1 might be through restoration of prosurvival PI3K/Akt/S6K signaling. These data shed some light on the treatment of intractable DNP and suggested that IGF-1 might be a potential effective agent on relieving high glucose induced neurotoxicity.

Nonagenarians with acute ischemic stroke tend to have a higher mortality and morbidity than younger patients. Tirofiban is a glycoprotein IIb/IIIa antagonist that has a therapeutic potential for ischemic stroke. Here, we provide a case report of a 93-year-old male patient with acute ischemic stroke. He presented with right-sided hemiparesis for 2 hours (National Institute of Health Stroke Scale, NIHSS = 23). Immediate treatment with endovascular tirofiban infusion achieved an improvement of intracranial blood flow and a progressively decreased NIHSS one day after admission (NIHSS = 16) and then seven days after admission (NIHSS = 7). After a follow-up of 90 days, the modified ranking score was 2. This case report suggests that endovascular application with tirofiban may be a favorable option for treating nonagenarians presenting with acute ischemic stroke and warrants further study.

Objective This study aims to observe the curative effect of Huaiqihuang Granules adjuvant therapy on primary nephrotic syndrome (PNS). Methods A total of 112 children with PNS were randomly divided into three groups, and changes in serum inflammatory cytokines, interleukin, lymphocyte subsets and immunoglobulin were observed. Results Before treatment, IL-18, TNF-α, CD8+ increased, while IL-10, CD4+, NK cells, IgA, IgG and Foxp3+Treg cells decreased. After Huaiqihuang Granules treatment, IL-18, TNF-α, CD8+ decreased, while IL-10, CD4+, NK cells, IgA, IgG and Foxp3+Treg cells increased. Conclusion Functions of cell immunity and humoral immunity in PNS patients before treatment were suppressed and disordered. Huaiqihuang granules can play a role in immunoregulation, with slight side reactions.

Tumor necrosis factor (TNF)-α-induced protein-8-like 2, or TIPE2, is a newly found immune negative regulatory molecule. This study further investigated the role of TIPE2 on proliferation and invasion of cervical squamous cancer cells. Expression of TIPE2 was compared in cervical squamous cancer tissues and adjacent normal tissues by Western blot and immunohistochemistry (IHC). Cervical squamous cancer cell lines, SiHa and C33A, were transfected with recombinant plasmid encoding TIPE2 and tested for cytologic characteristics. The impact of TIPE2 on phosphorylation of extracellular signal-regulated kinase (Erk) signaling pathway was also tested by Western blot analysis of key factors. TIPE2 expression was higher in cervical cancer tissues than that in normal tissue. IHC score of tumor tissue was negatively associated with lymphatic metastasis. Over expression of TIPE2 effectively inhibited the proliferation of cervical cancer cells. Wound healing and transwell assay showed that over expression of TIPE2 suppressed cell migration and invasion in vitro. Meanwhile, phosphorylation of Erk1/2 and upstream mitogen-activated protein kinase kinase (MEK) 1/2 was reduced by TIPE2. TIPE2 is negatively related with development of cervical squamous cancer. TIPE2 is an inhibitory factor of proliferation and invasion of cervical squamous cancer cells, probably through inhibiting Erk signaling pathway.

A 23-year-old female patient was referred for treatment of a posterior mediastinal tumour. There was no history of hypertension or headache and no other complaints. The patient’s blood pressure increased to 210/125 mmHg after surgically manipulating the tumour, subsequently reversing to severe hypotension (25/15 mmHg) immediately after the tumour was removed. The life-threatening and irreversible blood pressure drop was difficult to treat with fluid and vasopressors, and the patient ultimately died of cardio-respiratory failure. Asymptomatic paraganglioma can be non-functional but can also be fatal. For any lump in the thoracic cavity, paraganglioma should be ruled out.

An adenocarcinoma of the appendix invading the urinary bladder, which is difficult to be diagnosed before the operation, is an extremely rare disease. Only a few cases have been reported. Here we reported a case of patient diagnosed with the mucinous adenocarcinoma of the appendix invading the urinary bladder. The case reported in this study was a 54-years old man who was admitted due to a 6-month history of intermittent episodes of irritative voiding symptoms of the bladder, and weight loss. The patient did not have any gastrointestinal symptoms. The physical examination, laboratory examination, cytology of the urine, computed tomography and cystoscopy were inconclusive. The partial cystectomy, subsequent exploratory laparotomy and intraoperative frozen analysis revealed the appendiceal mucinous adenocarcinoma with a fistula to the urinary bladder. The appendectomy and the right hemicolectomy with a ileocolic anastomosis, the lymphadenectomy and the partial cystectomy limited to the anterior wall was performed. Six months after operation, the patient was in a good health with no obvious discomfort, no recurrence or distant metastases. The recommended treatment for the adenocarcinoma of the appendix invading the bladder with a fistula formation is as follows: appendectomy, right hemicolectomy with ileocolic anastomosis, lymphadenectomy, partial cystectomy and intraperitoneal hyperthermic chemoperfusion.

Backgroud The aim of this study was to evaluate the therapeutic effect of minocycline on treating experimental subarachnoid hemorrhage (SAH) in rats and to explore its possible molecular mechanism. Methods SAH was induced in male Sprague-Dawley rats by endovascular perforation. The rats were treated with minocycline (25 mg/kg or 50 mg/kg) or saline at 2 hand 12 h post SAH. Neurological function, cerebral hemorrhage, and edema were scored at 48 h post SAH. Cell death and P2X4 receptor (P2X4R) expression were observed in the prefrontal cortex (PFC). Results Treatment with a highdose of minocycline significantly improved the neurological function score, and attenuated cerebral hemorrhage and edema. Low-dose minocycline could reduce hemorrhage, but the effect on neurological deficits and brain edema was not obvious. Minocycline treatment could alleviate neuronal apoptosis in the PFC, which was related to reduced expression of inflammatory cytokines. Immunofluorescence showed that P2X4R on microglia was activated after SAH. Minocycline treatment inhibited P2X4R activation and further suppressed the phosphorylation of downstream p38 MAPK. Conclusions Minocycline plays a neuroprotective role by attenuating early brain injury after experimental SAH. The therapeutic mechanism of minocycline may be mediated by the inhibition of P2X4R on microglia.

Objective This study aims to investigate the effect of rosuvastatin on sympathetic neural remodeling of the left atrium (LA) in rats after myocardial infarction (MI). Methods Rats were randomly divided into a three groups: sham group, statin group, and MI group. The mRNA expression levels of the growth-associated protein-43 (GAP43) and nerve growth factor (NGF) were measured by RT-PCR. Immunohistochemistry was used to detect the distribution and density of GAP43- and NGF-positive nerves. The expression levels of these proteins were quantified by Western blot. Results Compared with the sham group, the average optical density (AOD) values of GAP43 and nerve growth factor (NGF)-positive substances in the LA in the statin and MI groups were significantly higher ( P <0.01), but the AOD values in the statin group were lower than of those in the MI group ( P <0.01). Furthermore, the AOD values of GAP43 and NGF positive nerves in the left stellate ganglion in the statin and MI groups were significantly higher ( P <0.01), but the AOD values in the statin group were lower, when compared with the MI group ( P <0.01). Conclusion Rosuvastatin could effectively improve the sympathetic neural remodeling of LA in MI rats.

Objective The aim of this study is to investigate the protective effects of emodin (EMD) on the lung injuries in the rat models of liver fibrosis. Methods Liver fibrosis was established in rats and the effect of intervention using EMD treatment was determined. Liver and lung weight coefficients were measured and lung content of TNF-α (tumor necrosis factor α), MDA (malondialdehyde), NO (nitric oxide), and ONOO- (peroxynitrite) were determined. Finally, histopathological changes were evaluated. Results Compared with the normal control group, the lung weight coefficient was significantly increased in the fibrosis model group. Moreover, pulmonary edema and inflammatory responses were observed. Levels of TNF-α, MDA, NO, and ONOO- in the lung homogenate were significantly increased in the fibrosis model group. After EMD treatment, the lung weight coefficients were significantly reduced. Moreover, pathological changes in the lung tissue were dramatically alleviated. Levels of TNF-α, MDA, NO, and ONOO- were significantly decreased. Conclusion EMD exhibits protective effects against lung injuries in a rat model of liver fibrosis.

Background Transforming RhoA proteins (RHOA) and their downstream Diaphanous homolog 1 proteins (DIAPH1) or mDia1 participate in the regulation of actin cytoskeleton which plays critical role in cells, i.e., morphologic changes and apoptosis. Methodology To determine the cell viability the real time cell analysis (RTCA) and flow cytometry were used. To perform proteomic analysis, the label-free quantitative method and post-translation modification by the nano-HPLC and ESI-MS ion trap mass analyser were used. Results The results of the cell viability showed an increase of dead cells (around 30 %) in MCF-7/DOX-1 (i.e., 1μM of doxorubicin was added to MCF-7/WT breast cancer cell line) compared to MCF-7/WT (control) after 24 h doxorubicin (DOX) treatment. The signalling pathway of the Regulation of actin cytoskeleton (p<0.0026) was determined, where RHOA and mDia1 proteins were up-regulated. Also, post-translational modification analysis of these proteins in MCF-7/DOX-1 cells revealed dysregulation of the actin cytoskeleton, specifically the collapse of actin stress fibbers due to phosphorylation of RHOA at serine 188 and mDia1 at serine 22, resulting in their deactivation and cell apoptosis. Conclusion These results pointed to an assumed role of DOX to dysregulation of actin cytoskeleton and cell death.

Background Clostridium difficile infection (CDI) is the leading cause of nosocomial diarrhea. Co-colonization of key bacterial taxa may prevent the transition from asymptomatic C. difficile colonization to CDI. However, little is known about the composition of key bacterial taxa in asymptomatic patients. Methods In the present study, the culture method was used to examine the composition of stool microbiota in two asymptomatic patients from Guizhou, China. Results A total of 111 strains were isolated and phylogenetic relationships were determined by 16S ribosomal gene sequencing and Molecular Evolutionary Genetics Analysis version 7. The results demonstrated that Escherichia (33.3%, 37/111), Clostridium (24.3%, 27/111) and Enterococcus (11.7%, 13/111) exhibited a high ratio in asymptomatic patients. These isolates derived from two phyla: Firmicutes (51.3%, 57/111) and Proteobacteria (44.1%, 49/111). In addition, co-colonization of human pathogens Fusobacterium nucleatum , Ralstonia pickettii , Klebsiella pneumoniae , Klebsiella quasipneumoniae and Clostridium tertium with C. difficile was identified. To the best of our knowledge, these pathogens have not been co-isolated with C. difficile previously. Conclusions In summary, the present study identified the composition of fecal microbiota in two asymptomatic patients in Guizhou, China. These results suggested that co-infection with human pathogens may be ubiquitous during CDI progression.

Objective Periodontal disease (PD) is viewed today as multifactorial problems initiated and sustained by bacteria but significantly modified by the body’s response to bacterial plaque. Recent studies have suggested that gene polymorphisms could be involved in the pathophysiology of periodontitis. This study aimed to investigate a possible correlation of the polymorphisms of intercellular adhesion molecule-1 ( ICAM-1 ) and vascular cell adhesion molecule-1 ( VCAM-1 ) with PD. Methods The genotypes of ICAM-1 and VCAM-1 were initially determined in PD patients using denaturing high performance liquid chromatography (DHPLC). ELISA was then conducted to measure ICAM-1 and VCAM-1 protein levels. Next, the association of ICAM-1/VCAM-1 genotype distribution and expression with clinical indicators and severity of PD was analyzed. Results PD patients contained increased levels of hemoglobin A1c (HbA1c), total cholesterol (TC), triglyceride (TG), and low-density lipoprotein (LDL), increased ICAM-1 and VCAM-1 protein levels, and decreased high-density lipoprotein (HDL) level. The GG genotype and G allele at ICAM-1 rs5498, as well as the AG and GG genotypes and G allele at VCAM-1 rs3181092 may reduce PD risk. Conclusion To sum up, the overexpressed ICAM-1 and VCA M-1 as well as A allele of ICAM-1 rs5498 and VCAM-1 rs3181092 is associated with the onset of PD.

Human cystic echinococcosis is a zoonosis caused by the larval cestode Echinococcus granulosus . Clear cell renal carcinoma is the most common pathological type of renal cell carcinoma. Echinococcosis complicated with carcinoma is rarely reported. Here, we reported a female patient with echinococcal cyst of the liver accompanied with clear cell renal carcinoma. This 27-year-old woman was admitted for abdominal pain. The serological testing of hydatid cyst was positive and levels of tumor markers were within the normal range. The computed tomography and histological findings confirmed hepatic echinococcal cyst complicated with renal carcinoma of kidney. Preoperative liver function was grade A. The patient underwent pericystectomy of liver hydatid cyst and partial nephrectomy. No recurrence was found at 1 year of follow-up. Liver hydatid complicated with renal cell carcinoma is rare, which should be differentiated from liver metastasis of renal cancer. Surgical resection is the optimal treatment. This case may provide insight for the diagnosis and research on the co-occurrence of tumor and hydatid cyst.

This study aims to investigate the changes of aquaporin-4 (AQP4), β-amyloid precursor proteins (APP) and β-amyloid (Aβ) in brain tissues after cerebral ischemiareperfusion injury (CIRI), and evaluate the effect of edaravone. The Middle Cerebral Artery Occlusion was used to establish CIRI in rats. Rats were divided into control, model and edaravone groups. The neurological deficits in the model group were obvious and the neurological score increased compared to the control group, while the neurological deficits of the edaravone group were improved as the neurological score decreased compared to the model group. The number of pyramidel cells in the hippocampus of the model group was significantly decreased whereas edaravone could reverse this decrease. The model group had significantly higher levels of Aβ, APP and AQP4 than the control group and edaravone group, suggesting that they might be involved in the neuronal cell damage. Meanwhile, the increased AQP4 might enhance the permeability of cells, and thus cause cell damage and neurological deficit. Conclusively, edaravone could reduce brain edema, protect neuronal cells and improve the neurological impairment of rats possibly by decreasing the expression of Aβ, APP and AQP4. Therefore, edaravone may have the potential to treat neurodegenerative diseases (such as Alzheimer's disease).

Neutrophil is a key element in inflammation and stress disease, which are associated with poor clinical outcomes in various cardiac diseases. However, the clinical availability of neutrophil in patients with ST-elevation myocardial infarction (STEMI) and chronic renal dysfunction has not been known. Accordingly, we designed this retrospective cohort study to evaluate the differences of major adverse cardiovascular events incidence between renal dysfunctional STEMI patients with normal and high neutrophil levels. The primary end point was all-cause mortality. We analyzed 377 consecutive STEMI patients with chronic renal dysfunction. The results showed that during 12-48 months follow-up, death from any-cause occurred in 1.4% patients (4 of 290) in normal-level neutrophil group, as compared with 3.4% in high-level neutrophil group (3 of 87) (hazard ratio, 2.174 95% confidence interval, 1.024-10.248; P = 0.025). Kaplan-Meier survival analysis showed that there were significant differences between the two groups with respect to the risk of death (P=0.018), and heart failure (P=0.037).

Objective To explore the potential role of GSG2 in breast cancer progression. Methods The mRNA expression, DNA copy number and clinical data used in this study were obtained from the TCGA data portal. The copy number variations (CNVs) thresholds were determined according to the set of discrete copy number calls provided by Genomic Identification of Significant Targets in Cancer (GISTIC). Results The mRNA expression level of GSG2 in 112 breast cancer tissues was much higher than that in adjacent normal tissues. GSG2 was significantly upregulated in stage II compared with stage I, and there was no differential expression of GSG2 between tumors with or without metastasis. Heterozygous deletion occupied 57.1% of CNVs for GSG2 gene in breast cancer samples. Patients with higher GSG2 expression tended to suffer from poorer prognosis. Conclusion Our profiling analysis indicated the overexpression of GSG2 might play an important role in breast cancer development, suggesting that GSG2 could be a new target for breast cancer treatment, making GSG2 inhibitors becoming potential drugs for breast cancer therapy.

Objective This study investigatesthe nuciferine capacity to regulate the liver’s lipid metabolism regarding steatosis and injury in STZ-induced diabetic rats. Materials and Methods The rats were randomly divided into groups control, diabetic and nuciferine 200 mg/kg/ day treatment. After 4 days of STZ injection, the nuciferine group was treated and administered via oral gavages for 4 weeks. At the end of experiment, blood, liver, myocardial and muscular samples were collected. Results Nuciferine-treated significantly increased the body weight from 339.4g to 367.8g, but significantly decreased the food and water intake compared with diabetic rats. Also, the nuciferine-treated rats had significantly decreased TC, TG, and FFAs in the liver compared with the diabetic group, especially the serum markers of blood glucose. These were associated with the gene expression related to lipogenesis which was significantly down-regulated; the gene expression involved in lipolysis and fatty acid β-oxidation was significantly up-regulated. Discussion and Conclusion The data provide evidence that nuciferine supplementation could protect the liver by regulating lipid metabolism gene expression resulting in decreasing the steatosis and injury in diabetic rat. Thus, nuciferine could be developed as a diabetic adjuvant food additive in future.

Objective To analyze the expression and its clinical significance of the SEC24D gene in breast cancer. Methods The dataset of breast cancer were searched in the UALCAN database, and the data obtained were mined and combined with literature analysis. Results The mRNA expression of the SEC24D gene in breast cancer tissues was significantly higher than that of breast normal tissues from the UALCAN database ( P < 0.05). The promoter methylation levels of the SEC24D gene in breast cancer tissues were lower than that of breast normal tissues ( P < 0.05). Survival analysis showed that the relapse-free survival of breast cancer patients with a higher expression of SEC24D gene was significantly worse than those patients with a lower expression of SEC24D ( P < 0.05). Conclusion The SEC24D gene has a high expression in breast cancer tissues and its expression level was related to the prognosis of breast cancer patients.

Tendon-bone healing after injury is an unsolved problem. Several types of stem cells are used as seed cells. However, the optimal co-culture ratio of different types of cells suitable for tissue engineering as well as the stimulator for facilitating the differentiation of stem cells in tendon-bone healing is unclear. In this study, the proliferation of both bone marrow-derived stem cells (BMSCs) and tendon stem cells (TSCs) was increased at a 1:1 co-cultured ratio, and proliferation was suppressed by Tenascin C (TNC). TNC treatment can promote osteogenesis or chondrogenesis of both BMSCs and TSCs under a 1:1 co-cultured ratio. In addition, the expression level of Rho-associated kinase (ROCK) increased in the process of TNC-induced osteogenesis and decreased in the process of TNC-induced chondrogenesis. Furthermore, the level of insulin-like growth factor 1 receptor (IGF-1R) and mitogen-activated protein kinase (MEK) was upregulated during the osteogenesis and chondrogenesis of both BMSCs and TSCs after TNC treatment. Although our study was conducted in rats with no direct evaluation of the resulting cells for tendon-bone healing and regeneration, we show that the proliferation of BMSCs and TSCs was enhanced under a 1:1 co-cultured ratio. TNC has a significant impact on the proliferation and differentiation of co-cultured BMSCs and TSCs. IGF-IR, ROCK, and MEK may become involved in the process after TNC treatment.

The giant panda (GP) was the most endangered species in China, and gut microbiota plays a vital role in host health. To determine the differences of the gut microbiota among the male, female and pregnant GPs, a comparative analysis of gut microbiota in GPs was carried out by 16S rRNA and ITS high-throughput sequencing. In 16S rRNA sequencing, 435 OTUs, 17 phyla and 182 genera were totally detected. Firmicutes (53.6%) was the predominant phylum followed by Proteobacteria (37.8%) and Fusobacteria (7.1%). Escherichia/Shigella (35.9%) was the most prevalent genus followed by Streptococcus (25.9%) and Clostridium (11.1%). In ITS sequencing, 920 OTUs, 6 phyla and 322 genera were also detected. Ascomycota (71.3%) was the predominant phylum followed by Basidiomycota (28.4%) and Zygomycota (0.15%). Purpureocillium (4.4%) was the most prevalent genus followed by Cladosporium (2.5%) and Pezicula (2.4%). Comparative analysis indicated that the male GPs harbor a higher abundance of phylum Firmicutes than female GPs with the contribution from genus Streptococcus . Meanwhile, the female GPs harbor a higher abundance of phylum Proteobacteria than male GPs with the contribution from genus Escherichia/ Shigella . In addition, the shift in bacteria from female to pregnant GPs indicated that phylum Firmicutes increased significantly with the contribution from Clostridium in the gut, which may provide an opportunity to study possible associations with low reproduction of the GPs.

Regeneration and repair with scarring of the skin are two different responses to tissue injury that proceed depending on the animal species. Several studies in multiple organisms have shown that the effectiveness of tissue repair gradually decreases with age in most vertebrates, while the molecular and cellular mechanisms underlying the diverse potentials remain incompletely understood. It is clear, however, that immune system actively participates in the whole process and immune-related activities can mediate both negative and positive roles to influence the quality and diversity of tissue response to damage. Compared with innate immunity, our understanding of the significance of adaptive immune cells in normal repair outcome is limited and deserves further investigation. Here, experimental evidence supporting the contribution of lymphocytes and the involvement of lymphoid organs in skin wound healing are discussed, focusing on the findings emerged in adult amphibians, key animal models for tissue repair and regeneration research.

In sole aquaculture production, consistency in the quality of produced eggs throughout the year is unpredictable. Hox genes have a crucial role in controlling embryonic development and their genetic variation could alter the phenotype dramatically. In teleosts genome duplication led paralog hox genes to become diverged. Direct association of polymorphism in hoxa1a , hoxa2a & hoxa2b of Solea solea with egg viability indicates hoxa2b as a potential genetic marker. High Resolution Melt (HRM) analysis was carried out in 52 viable and 61 non-viable eggs collected at 54±6 hours post fertilization (hpf). Allelic and genotypic frequencies of polymorphism were analyzed and results illustrated a significantly increased risk for non-viability for minor alleles and their homozygous genotypes. Haplotype analysis demonstrated a significant recessive effect on the risk of non-viability, by increasing the odds of disrupting embryonic development up to three-fold. Phylogenetic analysis showed that the paralog genes hoxa2a and hoxa2b , are separated distinctly in two clades and presented a significant ω variation, revealing their diverged evolutionary rate.

The aim of the study was to determine the role of dogs as a potential reservoir of Campylobacter spp. At the next stage of the research the frequency of occurrence of selected virulence genes, i.e. cadF , flaA and iam as well as genes responsible for the formation the cytolethal distending toxin (CDT), i.e. cdtA , cdtB and cdtC was determined. The isolates obtained in the research were tested for their resistance to selected antibiotics: ciprofloxacin (CIP), enrofloxacin (EF), erythromycin (E) and tetracycline (TE). Campylobacter spp. was found in 63 (12.6%) out of a total number of 500 isolates. 61 (12.2%) isolates were identified as C. jejuni . The number of C. jejuni isolates found in the younger animals was smaller ( p <0.05) than in the older ones. The frequency of occurrence of virulence genes and the genes responsible for the formation of CDT was significantly ( p <0.05) higher in the older dogs. A comparison of the effect of antibiotics showed that the isolates obtained from both age groups exhibited low resistance to erythrosine (13.5% in the group aged under 1 year and 8.6% in the group aged over 1 year). Both groups exhibited the highest resistance to ciprofloxacin and enrofloxacin.

The present study was performed to investigate the effects of dietary supplementation with Lactobacillus plantarum (CGMCC1.557) on egg production and fecal microbiota composition in laying hens. Sixty Hy-Line Brown laying hens (18 weeks old) were randomly divided into two groups. The control group was fed a basal diet only, and the test group was fed basal diet supplemented with a final concentration of 1.0 × 10 9 CFU/mL during the 10-week experimental period. Egg production and fecal microbiota composition were both assessed in 28-week-old hens using high-throughput sequencing technology. The results showed that, compared with the control group, the test group exhibited increased laying and feed intake rates ( p < 0.05). At the genus level, Lactobacillus was more abundant in the test group compared with the control group ( p < 0.05). Conversely, Romboutsia was more abundant in the control group compared with the test group ( p < 0.05). This study provides us with an insight into the potential use of L. plantarum as a food supplement in the laying hen industry. the study also provides us with a better understanding of the interplay between L. plantarum and the fecal microbiota of laying hens.

Ten leading wheat cultivars originating from the Plant Breeding and Acclimatization Institute (IHAR) - National Research Institute (Poland) and the Department of Gene Bank (Czech Republic) were used to establish a field experiment in 2017 and 2018 at the Dłoń Experimental Farm. The analyzed wheat genotypes were characterized by diversified field resistance to leaf rust. Jubilatka, Thatcher and Sparta were the most resistant cultivars in field conditions in both 2017 and 2018. The aim of the work was to identify the Lr11, L13, Lr16 and Lr26 genes encoding resistance to leaf rust using molecular SSR markers ( wmc24, wmc261, Xgwm630, Xwmc764 and P6M12 ) and to develop multiplex PCR conditions to accelerate identification of these genes. Markers of three leaf rust resistance genes have been identified simultaneously in these cultivars. Jubilatka, Thatcher and Sparta cultivars may serve as a good source of the analyzed leaf rust resistance genes. In addition, multiplex PCR conditions have been developed for the simultaneous identification of the Lr11 and Lr16 and Lr11 and Lr26 gene pairs.

This study was carried out to determine the effects of some additives on the potential nutritional value and silage quality of guar bean (Cluster bean = Cyamopsis tetragonoloba ) silages. It was hypothesized that the use of cereal grains, molasses and ecomass will reduce silage fermentation pH by increasing lactic acid production and positively improve CP content and silage quality. Four different silage groups were established; (control, grain (5%), molasses (10%) and ecomass+molasses (10%+5%)). Fresh guar beans were thoroughly mixed with the additives to homogenize, then ensiled and opened after 60 days. The results of this study revealed that guar bean silages could be used as an alternative forage in ruminant feeding because of its higher protein content (13.88%), forage feed value and silage quality. The use of molasses (GSM) and molasses + ecomass (GSEM) as additives has significantly (P ≤0.01) increased the silage quality and feed value compared to the control group. It was observed that GSM and GSEM silage groups had the highest values in terms of lactic acid content. In conclusion, guar silage can be used as an alternative feed for ruminants, but the doses of barley and molasses should be correct and tested in in vivo studies.

Potato juice (PJ), commonly considered a burdensome waste, is rich in various compounds with bioactive properties. It has long been considered a remedy for gastric problems in traditional folk medicine. If valorization of PJ through implementation in the production of functional foods is to be considered, stabilization methods must be developed to allow long-term storage of this seasonal product. It is important that such methods are chosen with regard to their effect on the bioactive value of the obtained product. In this study, the impact of four stabilization methods on the antioxidant and cytotoxic activities of PJ was investigated. Elevated temperatures were used in thermal deproteinization used to obtain DPJW (deproteinated potato juice water) and spray-drying of FPJ (fresh potato juice) that resulted in SDPJ. Freeze drying and cryoconcentration were the low temperature processing methods that yielded PJL (potato juice lyophilisate) and CPJ (cryocorncentrated potato juice), respectively. All processed materials were characterized chemically and compared with raw materials in terms of phenolic compounds content, antioxidant activity as well as cytotoxicity to human tumor cells isolated from the gastric mucosa (Hs476T cell line), colon (Caco-2 and HT-29 cell lines), and normal cells isolated from the small intestine and colon epithelium (IEC-6 and NCM460 cell lines). It was stated that high-temperature processes – thermal deproteinization and spray-drying – yielded products with increased antioxidant potential (TEAC) that also showed increased cytotoxic activity towards intestinal cancer cells. At the same time the cytotoxicity towards normal cells remained on par with that of fresh PJ (IEC-6 cells) or decreased (NCM460 cells). Thermal deproteinization significantly decreased the content of glycoalcaloids in the juice, while spray drying did not have such an effect. The two low-temperature processes investigated – cryoconcentration and freeze drying – did not affect the PJ cytotoxic activity towards any of the cell lines used in the tests, whereas they did affect the antioxidant properties and glycoalcaloids content of PJ.