Volume 50, Issue 6

Kidney diseases are a worldwide public health problem with more than 850 million patients all over the world. Therefore, early diagnosis is essential for effective treatment of any renal disease. Most biomarkers have the potential to be used in diagnosis, screening, prognosis, and follow-up, but it is challenging to confirm associations with certain diseases. Although some biomarkers such as urea and creatinine are used in clinical practice for kidney diseases, these molecules’ sensitivity and specificity are not at the desirable level. Pregnancy-associated plasma protein-A (PAPP-A) is being used for the screening of Down’s syndrome in the first trimester of pregnancy, however, increased expressions have been reported in various kidney diseases. This study aimed to evaluate the clinical significance of PAPP-A as a potential biomarker for diagnosing and monitoring renal diseases. We searched Pubmed and Web of Science databases using PAPP-A, kidney diseases, acute kidney injury, chronic kidney disease, hemodialysis, peritoneal dialysis, kidney transplant, diabetic nephropathy, polycystic kidney disease, and kidney cancer. According to our search, PAPP-A seems to be a candidate biomarker for diabetic nephropathy and chronic kidney disease, yet further studies are needed to detect diseases in the early stage.

Objectives Forkhead box (FOX) transcription factors and long non-coding RNAs (lncRNAs) localize within tumor cell nuclei in various cancers, suggesting their involvement in transcriptional regulation. This study aimed to investigate expression changes of FOX family members and selected lncRNAs to explore their potential regulatory roles in differentiated thyroid cancer (DTC). Methods Expression levels of FOX genes and lncRNAs were analyzed using RT-PCR, and protein levels were measured via ELISA in patients diagnosed with DTC who received radioactive iodine (RAI) therapy. Results RT-PCR analysis revealed significant differences in FOXO1 and HOXA11-AS expression before and after RAI therapy (p<0.05). FOXO3 , FOXP1 , FOXP2 , and CASC21 showed significant expression differences in pre-RAI samples, while MEG3 and KCNQ1OT1 differed significantly post-RAI (p<0.05). Moreover, strong correlations were identified between FOXO3 , FOXP2 , and key lncRNAs including MEG3 , KCNQ1OT1 , CASC21 , and HOXA11-AS (p<0.05). ELISA results showed a significant change in FOXP1 protein levels, and TSH and Tg levels differed significantly across treatment stages (p<0.05). CASC21 exhibited the highest AUC in ROC analysis. Conclusions Despite the limited sample size, the findings suggest the potential of these genes, particularly CASC21, as biomarkers in DTC. These results may contribute to enhancing RAI therapy efficacy and advancing personalized treatment approaches.

Objectives The capacity to sense and respond to cellular energy stress is an important factor in tumorigenesis. We aimed to investigate the proliferation patterns of KRAS-mutant NSCLC cell lines (A549, Calu-1, H2009) under high (HG) and low glucose (LG) conditions w/wo DCA, which alters metabolic pathways and promotes oxidative phosphorylation. Methods Cell viability was detected by MTT assay, protein expressions were determined by Western Blot and RNA expressions were analyzed by RT-qPCR. Results NSCLCs exhibited lower proliferation in HG compared to LG conditions. The effect of DCA in reducing the proliferation of the cells, was higher in HG compared to LG condition. We detected lower pRB levels in LG and in DCA treated conditions. LG conditions also showed higher p21 levels w/wo DCA while AMPK activation was more pronounced in 48 h. p-JAK2 was dominant in conditions having LG and p-Stat3 activation varied between all cells. The eIF4e activation was induced in LG w/wo DCA in all the cells. Despite p21-mediated cell cycle arrest, eIF4e and JAK2 signaling promoted proliferation. Conclusions Our results clearly showed the potential for increased aggressiveness and the delicate balance of cellular metabolic adjustments in response to energy stress in KRAS-mutant NSCLC cells.

Objectives Cervical cancer (CC) is a malignant tumor that poses a serious threat to women’s lives and has a complex pathogenesis. This study aims to investigate the mechanism underlying the lncRNA CDKN2B-AS1 in CC. Methods The expression of CDKN2B-AS1, hsa-miR-134-5p, and CCND1 was detected by RT-qPCR and western blotting, and their targeted regulatory relationships were verified by dual-luciferase reporter assay and overexpression/knockdown assay. CCK-8, Transwell, and Annexin V-FITC/PI staining experiments were used to assess the proliferation, migration, invasion and apoptosis of CC cells (HeLa and SiHa), respectively. The Kaplan-Meier survival curve was used to evaluate the clinical value of CDKN2B-AS1 in CC. Results CDKN2B-AS1, CCND1 were high-expressed, and hsa-miR-134-5p was low-expressed in CC cells and tissues. Silencing CDKN2B-AS1 significantly inhibited the growth and metastasis of HeLa and SiHa cells, and induced their apoptosis. The overall survival time of CC patients with high CDKN2B-AS1 was shorter (log rank p<0.05). Hsa-miR-134-5p mimic attenuated the luciferase activity of cells transfected with wild-type CDKN2B/CCND1 luciferase vector. Overexpression of CDKN2B-AS1 decreased the intracellular level of hsa-miR-134-5p, while its knockdown was inverse. The inhibition of hsa-miR-134-5p promoted CCND1 expression. Conclusions High CDKN2B-AS1 level predicted a poor prognosis of CC patients, and it promoted CC development by down-regulating hsa-miR-134-5p to enhance CCND1 expression.

Objectives Research has shown that miRNAs play an essential role in the pathophysiology diabetic kidney disease (DKD). This study primarily explored the expression of miR-709 in DKD and elucidated its potential clinical significance for patients with DKD. Methods A total of 95 patients with DKD were included in the case group, while 102 T2DM patients constituted the control group. 50 healthy individuals were selected as the healthy group. Real-time quantitative PCR was employed to assess the expression levels of miR-709. The receiver operating characteristic curve was used to evaluate the diagnostic value of miR-709 in DKD. Spearman correlation analysis was performed to explore the relationship between serum miR-709 levels and various clinical data variables. Furthermore, an in vitro DKD cell model was developed to investigate the effect of miR-709 expression in DKD. Results In DKD patients, serum miR-709 levels are markedly reduced and have high diagnostic value. MiR-709 associated with clinical features and inflammation in DKD. Up-regulation of miR-709 significantly suppressed HG-induced proliferation and inflammatory factor levels in human mesangial cells (HMCs). Conclusions The decreased level of miR-709 is closely linked to DKD and its inflammation, which is expected to become a potential biomarker for clinical screening and a target for therapeutic intervention.

Objectives Obesity is a global health problem related to reduced life quality and shorter life expectancy. Given the fact that genetic factors play a role in obesity development, studies have suggested that polymorphisms of cannabinoid receptor 1 ( CNR1 ), monoacylglycerol lipase ( MGLL ), and fatty acid amide hydrolase ( FAAH ) genes located in the endocannabinoid system (ECS) are related to overweight/obesity risk. Accordingly, we aimed to assess the genetic susceptibility to overweight/obesity in individuals with Turkish ancestral origin concerning these ECS genes. Methods The present study comprised 194 unrelated individuals with Turkish ancestral origin living in the Marmara Region. The study population is categorized into two groups according to their body mass index (BMI) overweight/obese group (BMI≥25) and normal weight group (18.5<BMI<25). Genotyping of the CNR1 rs1049353 , MGLL rs60430 , and FAAH rs324420 polymorphisms was performed using the real-time polymerase chain reaction (RT-PCR). Results The GG genotype frequency of CNR1 rs1049353 was significantly higher in overweight/obese individuals than in normal weight individuals (75.5 vs. 61.4 %, respectively; p=0.034). No difference was observed in terms of MGLL rs604300 and FAAH rs324420 polymorphisms between the groups (p>0.05). The genotype distributions of CNR1 , MGLL , and FAAH genes were compatible with the Hardy–Weinberg equilibrium (p>0.05). Conclusions Our findings supported that individuals with AA/AG genotype of CNR1 rs1049353 have reduced overweight/obesity risk; however, neither FAAH rs324420 nor MGLL rs604300 showed any association. A relatively smaller sample size and lack of inclusion of obesity-related measurements could be considered the limitations of this study. Further research is needed to confirm or disprove our results.

Objectives Increased levels of homocysteine (Hcy) in the blood are considered to be a risk factor for atherosclerosis. Endothelial damage induced by Hcy can lead to deterioration of coronary, cerebral, and peripheral vascular beds, contributing to cardiovascular morbidity and mortality. However, the mechanisms underlying Hcy-induced damage have not been fully elucidated. This study aimed to investigate the effect of Hcy on low-density lipoprotein (LDL) oxidation and inducible nitric oxide synthase (iNOS) expression, both of which are thought to play a role in the pathogenesis of atherosclerotic plaque formation and regulation of inflammation. Methods L-929 fibroblast-like cells and RAW 264.7 macrophages were treated with varying concentrations of Hcy (50, 100 and 200 μg/mL for L-929 cells, 10, 25, 100, 200 μg/mL for RAW 264.7 cells) to assess LDL oxidation and iNOS expression using immunocytochemical and immunofluorescence methods. Images obtained from cell culture experiments were quantitatively analyzed using color and brightness histograms, followed by statistical evaluation. Results Hcy significantly and dose-dependently increased LDL oxidation and intracellular oxidized LDL (ox-LDL) accumulation in both cell groups. Hcy also significantly increased iNOS expression in both cell groups in a dose-dependent manner. Conclusions These findings suggest that Hcy may play a role in the pathogenesis of atherosclerosis and inflammatory processes. The increased levels of ox-LDL and the elevated iNOS expression, particularly in macrophages, may contribute to the exacerbation of atherosclerosis and related diseases.

Objectives Wilson’s disease (WD) leads to copper accumulation in the liver and brain. It generates oxidative stress, which can damage the liver and central nervous system (CNS). The concentrations of oxidized lipid products, such as prostaglandins and isoprostanes, in blood and urine can indicate both the presence and the extent of damage. However, there is limited research specifically investigating these oxidative stress biomarkers in WD, making this study a novel contribution to the field. Methods In this study, serum and 24-h urine samples were collected from a total of 64 individuals, including 22 WD patients with hepatic involvement (HI-WD), 17 with neurological-hepatic involvement (NHI-WD), and 25 healthy controls (HC). A liquid chromatography mass spectrometry (LC-MS/MS) method was developed and validated to quantify specific prostaglandins and isoprostanes in serum and urine samples. Results The results showed significant differences in the levels of iPF 2α -III, 2,3-dinor-iPF 2α -III, 5-iPF 2α -VI in serum as well as PGE 2 , PGD 2 , PGF 2α , iPF 2α -III and 5-iPF 2α -VI in 24-h urine samples between WD and HC groups. Notably, the concentration of PGD 2 in urine was significantly higher in the NHI-WD group compared to both the HC and HI-WD groups. Our novel urine biomarker panel has the potential to be used for grading disease severity, monitoring therapy and assessing CNS involvement in WD. Conclusions It represents one of the first to explore a panel of oxidative stress biomarkers, particularly prostaglandins and isoprostanes, in both serum and urine samples in WD patients, making it an original contribution to the current literature on WD.

Objectives Reflux esophagitis (RE) arises from the regurgitation of gastric fluid that contains acids, bile, and other damaging components, significantly impacting on the quality of life. This study aimed to investigate the functions of miR-15b-5p and AdipoQ receptor family member 3 (PAQR3) in reflux esophagitis, as well as to delve into the effect on innate immune response. Methods The study enrolled 54 patients with RE and 33 healthy subjects. qRT-PCR was used to detect the expression level of miR-15b-5p, PAQR3, TLR2, TLR4 and MyD88. The ELISA kits were employed to assess the alterations in the levels of IFN-β, IL-6, and IL-8. Additionally, a dual-luciferase reporter assay was used to verify the binding relationship between miR-15b-5p and PAQR3. Results miR-15b-5p was significantly elevated in RE patients. Cell model tests demonstrated that miR-15b-5p was upregulated in cells treated with acid medium. Inhibition of miR-15b-5p significantly suppressed the expression of innate immunity related genes induced by acid medium. miR-15b-5p targets and regulates PAQR3. In addition, miR-15b-5p activated innate immune responses by blocking PAQR3. Conclusions miR-15b-5p may be involved in the innate immune response of reflux esophagitis by regulating PAQR3.

Objectives The aim of this study was to investigate the impact of miR-26b-5p on diagnosis of bronchiolitis as well as its relationship with inflammatory factors. Methods This study conducts a retrospective study of infants and toddlers with bronchiolitis, coughing and wheezing as the main symptoms (RSV-positive and RSV-negative). The qRT-PCR was used to examine miR-26b-5p expression levels. Diagnostic accuracy of miR-26b-5p in infant and toddler respiratory diseases was assessed by ROC curve. Pearson correlation analysis was applied to demonstrate the relationship between miR-26b-5p and clinical severity markers, along with immune-related genes such as PTGS2 and TRAF3 . RSV-infected HBECs were used to validate the effect of miR-26b-5p on inflammatory factors. IL-6, IL-10 and TNF-α were measured by ELISA kits. Results MiR-26b-5p expression was reduced in infant and toddler respiratory diseases including bronchiolitis, coughing and wheezing. The ROC curve indicated that miR-26b-5p had a high diagnostic accuracy for differentiating respiratory diseases from healthy infants and toddlers. Notably, miR-26b-5p returned to baseline in infants and toddlers who had recovered from bronchiolitis. Pearson correlation analysis revealed a negative relationship of miR-26b-5p with PTGS2 and TRAF3 . RSV infection of HBECs reduced miR-26b-5p expression, and transfection with miR-26b-5p mimic reversed this trend. Furthermore, IL-6 and TNF-α levels were significantly elevated and IL-10 levels were reduced in RSV-infected HBECs, which was reversed by overexpression of miR-26b-5p. Conclusions MiR-26b-5p may be involved in disease progression through modulation of inflammatory responses and may be a diagnostic biomarker for infant and toddler respiratory diseases.

Objectives Dry eye disease (DED) is a prevalent ocular surface, and its pathogenesis remains inadequately elucidated. This research examined the possible regulatory mechanisms of lncRNA MALAT1 in DED to provide biomarkers for DED treatment. Methods qPCR was employed to assess the expression levels of MALAT1 and miR-302d in DED patients’ serum and HCECs cells. ELISA was utilized to measure the levels of IL-1β, IL-6, IL-8, and IFN-γ in DED patients’ serum and in various cell transfection groups. The effects of different cell transfection groups on the proliferation and apoptosis of HCECs were analyzed using the CCK-8 assay and flow cytometry. Furthermore, potential regulatory pathways were explored through GO and KEGG enrichment analyses of the target genes of miR-302d. Results MALAT1 exhibited a significant up-regulation in DED patients’ serum, while miR-302d was notable down-regulation, with a significant negative correlation between two molecules. MALAT1 was positively correlated with the Ocular Surface Disease Index (OSDI) and Symptom Assessment in Dry Eye (SANDE), suggesting its potential utility in evaluating the severity of DED. MALAT1 inhibited the proliferation of TNF-α-induced HCECs, promoted apoptosis, and enhanced the release of inflammatory factors. However, the suppression of miR-302d mitigated the impact of MALAT1 silencing on the function of HCECs. Enrichment analysis revealed that the target gene of miR-302d played a role in regulating cellular homeostasis and growth in HCECs. Conclusions The sponging effect of MALAT1 on miR-302d caused changes in HCECs cell function, and led to DED. MALAT1 was an important parameter reflecting the severity of DED patients.

Objectives Efforts to minimize pre-analytical errors remain significant in laboratory procedures. However, the existing literature has not thoroughly explored the awareness of errors that could influence the process before patients submit samples, and there is yet to be an internationally recognized standard on this matter. In this study, a set of 20 questions was posed to participants in the waiting area to assess their knowledge and awareness regarding the pre-analytical process. Methods In this study, voluntary participants were administered an awareness survey consisting of 4 open-ended and 16 multiple-choice questions. Four voluntary pollsters conducted a 20-item survey of voluntary individuals aged 18 and over who came to the blood sampling department of Pamukkale University Hospital to provide samples. The data obtained were analyzed using SPSS 25.0. Results The average age of the 328 participants was 30 ± 14 years, with 55.9 % of them being female. 68 % of the participants responded that they knew the rules before sampling. The % of those who believed fasting would not affect the sample result was 42.7. The 89 % of the participants mostly answered “yes” to the survey’s last question, which asked, “The questions directed to me showed that there may be points I do not know about blood sample giving”. Conclusions This article, which mentions assessing awareness and knowledge levels before blood sampling, highlights the necessity for education among healthcare professionals and patients.

Objectives Immunofixation electrophoresis (IFE) is a biochemical diagnostic method used in the diagnosis, treatment, and follow-up of monoclonal gammopathies. Paraproteinaemias involve overproduction of immunoglobulins, known as paraproteins or monoclonal proteins (M-proteins). M-protein can be detected as a sharp band on serum protein electrophoresis (SPE). Paraprotein types produced in monoclonal gammopathies can be detected by the IFE method. In our study, we aimed to retrospectively evaluate the types of paraproteinaemia detected by the IFE method in the laboratory and to compare our data with the literature. Methods A retrospective evaluation of the IFE results of a medical biochemistry laboratory of a tertiary care education and research hospital in Turkey in terms of the frequency and type of monoclonal gammopathy was performed in 2022. Serum free kappa/lambda (κ/λ) ratios and IFE results were compared. Results In our study, 1,169 serum IFE results were retrospectively analyzed, and paraprotein bands were detected in 116 different cases. The frequency of detected paraproteins was determined to be 10 %. A comparison between serum free κ/λ ratios and IFE results yielded 87 % concordance. Conclusions In our retrospective analysis, we obtained results similar to those reported in the literature in terms of serum immunofixation data, age, gender, and immunoglobulin types. We identified IgG-κ (20 %) as the most common paraprotein type. We believe that our study will contribute to the literature in terms of the frequency and variety of paraproteinemia. The κ/λ ratio is of limited use, since most abnormal IFE sera have normal free light chain (FLC) ratios.

Objectives Geniposide (GNP) is a natural compound that possesses both antioxidant and anti-inflammatory activities. Here, we examined GNP effect on obesity-induced cardiac injury in Wistar rats. Methods Wistar rats were grouped into control, high fat diet (HFD) and HFD+GNP. Levels of cardiac total antioxidants and malondialdehyde (MDA) were measured. TUNEL assay was applied to detect cardiac cells apoptosis. Gene expression analysis was performed for catalase (CAT), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), interleukin-10 (IL-10), Caspase-3 and -9, B-cell lymphoma-2 (Bcl2) and Bcl-2-associated X protein (Bax) by real time PCR and fold-change ratio was calculated using 2 −ΔΔCt method. Results GNP administration not only decreased hyperlipidemia, atherogenic coefficient (0.65±0.06 vs. 1.70±0.31; p<0.001) and cardiac risk ratio (1.65±0.23 vs. 2.70±0.29; p<0.01), but also it attenuated inflammatory cell infiltration and cardiomyocyte vacuolization, and cardiac apoptotic cells (8.22±1.05 % vs. 21.46±2.44 %; p<0.001) compared to HFD alone. GNP significantly increased cardiac TAC (578.11±36.2 μM/g tissue; p<0.01) and zinc (144.17±8.57 vs. 102.13±5.81 ng/g tissue; p<0.001) contents, but significantly decreased MDA content (3.02±0.5 vs. 4.71±0.38 nmol/mg protein, p<0.01). A significant trend was found toward overexpression of Caspase-8, Bax, Caspase-3 and TNF-α in HFD group, but IL-10, SOD, CAT and Bcl2 expression was significantly decreased. Conclusions GNP is a safe and effective natural product that protects obesity-induced cardiac injury by elevating total antioxidant pools, suppressing inflammation, oxidative stress and apoptosis.

Objectives This study aims to explore the therapeutic potential of Xanthohumol (XH) in mitigating osteoarthritis (OA) progression by inhibiting extracellular matrix (ECM) degradation. Specifically, it investigates the role of XH in reducing matrix metalloprotease-13 (MMP-13) activity and modulating the PRKR-like endoplasmic reticulum kinase/activating transcription factor 4 (PERK/ATF4) pathway while enhancing nuclear factor-erythroid 2-related factor 2 (Nrf2) activity. Methods In vitro experiments were conducted on OA chondrocytes (CTs) to evaluate MMP-13 production and collagen II expression following XH treatment. In vivo , XH was administered intraperitoneally, and cartilage degradation was assessed. Molecular analysis examined the expression of C/EBPβ, a downstream effector of the PERK/ATF4 pathway, and Nrf2 activity. Overexpression and knockdown experiments were performed to elucidate XH’s mechanistic effects. Results XH treatment reduced MMP-13 production by 46 % and increased collagen II expression by 38 % in vitro . In vivo , XH administration resulted in a 31 % decrease in cartilage degradation scores compared to controls. Mechanistically, XH decreased C/EBPβ expression and increased Nrf2 activity by 52 %. Overexpression of C/EBPβ or activation of the PERK/ATF4 pathway nullified XH’s protective effects, while Nrf2 knockdown abolished its benefits. Conclusions XH inhibits ECM degradation in OA by suppressing the PERK/ATF4/C/EBPβ pathway and activating Nrf2, demonstrating its potential as a low-cytotoxicity therapeutic agent for OA treatment. This approach offers a promising strategy for mitigating cartilage damage and advancing OA management.

Objectives Chemotherapy-induced senescent cells are recognized to lead to cancer progression and resistance to treatment by releasing factors associated with the senescence-associated secretory phenotype (SASP). Therefore, targeting SASP via senomorphic agents is an emerging therapeutic strategy. In this study, we evaluated the potential of L-type Ca 2+ channel blockers, nifedipine and verapamil, and K + ATP channel openers, minoxidil and diazoxide, to influence senescence development and SASP-related secretory activity. Methods Senescence development was evaluated by beta-galactosidase activity and senescent cell morphology. Senomorphic activity was evaluated by measuring the secretion levels of total protein and IL-6, which are considered indicators of the SASP. Additionally, the effects of SASP were tested for cancer cell proliferation using the transwell system and cell migration using wound healing methods. Results Our findings indicate that neither L-type Ca 2+ channel blockers, nifedipine and verapamil, nor K + ATP channel openers, minoxidil and diazoxide, influence the induction of senescence by doxorubicin or alter senescent cell morphology. The K + ATP channel openers minoxidil and diazoxide notably inhibit the secretion of the proinflammatory cytokine IL-6 and VEGF-A in senescent HeLa cells, implying potential senomorphic properties. Moreover, although minoxidil and diazoxide did not reduce the proliferative effects of SASP on cancer cells, these drugs modulated the secretory phenotype of senescent cells to reduce HeLa cell migration significantly. Conclusions We suggest that minoxidil and diazoxide may have potential as senomorphic adjuvants in cancer therapy. If confirmed by invivo and clinical studies, these drugs may be repurposed to attenuate SASP-induced tumor progression. Moreover, K + ATP channels represent potential targets for senotherapy.