Volume 41, Issue 1 -

Background A potential correlation between sex hormones, such as androgens and estrogens, and the development and progression of hepatocellular carcinoma (HCC) has been proposed. However, its details, in particular, aromatase status in diseased human liver has remained largely unknown. Materials and methods We immunolocalized aromatase, 17β-hydroxysteroid dehydrogenase (17β-HSD) type 1 and 17β-HSD type 2 in a total of 155 cases, consisting of normal liver (n = 10), nonalcoholic steatohepatitis (NASH) (n = 18), primary sclerosing cholangitis (PSC) (n = 6), primary biliary cholangitis (PBC) (n = 13), biliary atresia (n = 18), alcoholic hepatitis (n = 11), hepatitis C virus (HCV) (n = 31), HCV sustained virologic response (HCV-SVR) (n = 10), hepatitis B virus (HBV) (n = 20), HBV sustained virologic response (HBV-SVR) (n = 8) and infants (n = 10). Results Immunoreactivity scores of aromatase in HBV (59.5 ± 30.9), HBV-SVR (68.1 ± 33.5) and infants (100.5 ± 36.6) were significantly higher than those in normal liver (26.0 ± 17.1). Scores of 17β-HSD type 1 in any etiology other than HBV (116.3 ± 23.7) and infants (120.0 ± 28.5) were significantly lower than those in normal liver (122.5 ± 8.6). Scores of 17β-HSD type 2 in NASH (74.4 ± 36.6) were significantly lower than those in normal liver (128.0 ± 29.7). Conclusion High immunoreactivity scores of aromatase and 17β-HSD type 1 in the patients with HBV suggest a correlation between HBV infection and in situ estrogen synthesis in hepatocytes.

Hepatocellular carcinoma (HCC) is the most common type of liver cancer. HCC is particularly aggressive and is one of the leading causes of cancer mortality. In recent decades, the epidemiological landscape of HCC has undergone significant changes. While chronic viral hepatitis and excessive alcohol consumption have long been identified as the main risk factors for HCC, non-alcoholic steatohepatitis (NASH), paralleling the worldwide epidemic of obesity and type 2 diabetes, has become a growing cause of HCC in the US and Europe. Here, we review the recent advances in epidemiological, genetic, epigenetic and pathogenic mechanisms as well as experimental mouse models that have improved the understanding of NASH progression toward HCC. We also discuss the clinical management of patients with NASH-related HCC and possible therapeutic approaches.

The prevalence of non-alcoholic fatty liver disease (NAFLD) is increasing. Determining the pathogenesis and pathophysiology of human NAFLD will allow for evidence-based prevention strategies, and more targeted mechanistic investigations. Various in vivo, ex situ and in vitro models may be utilised to study NAFLD; but all come with their own specific caveats. Here, we review the human-based models and discuss their advantages and limitations in regards to studying the development and progression of NAFLD. Overall, in vivo whole-body human studies are advantageous in that they allow for investigation within the physiological setting, however, limited accessibility to the liver makes direct investigations challenging. Non-invasive imaging techniques are able to somewhat overcome this challenge, whilst the use of stable-isotope tracers enables mechanistic insight to be obtained. Recent technological advances (i.e. normothermic machine perfusion) have opened new opportunities to investigate whole-organ metabolism, thus ex situ livers can be investigated directly. Therefore, investigations that cannot be performed in vivo in humans have the potential to be undertaken. In vitro models offer the ability to perform investigations at a cellular level, aiding in elucidating the molecular mechanisms of NAFLD. However, a number of current models do not closely resemble the human condition and work is ongoing to optimise culturing parameters in order to recapitulate this. In summary, no single model currently provides insight into the development, pathophysiology and progression across the NAFLD spectrum, each experimental model has limitations, which need to be taken into consideration to ensure appropriate conclusions and extrapolation of findings are made.

Non-alcoholic fatty liver disease (NAFLD) is a clinical entity of high prevalence in the world characterized by fatty infiltration of liver tissue in the absence of alcohol consumption. The natural history of the disease develops in successive phases reflected in different histological stages, with 10–20% of patients developing liver cirrhosis and fibrosis. Fibrosis is a basic connective tissue lesion defined by the increase of the fibrillary extracellular matrix (ECM) components in a tissue or organ. Matrix metalloproteinases (MMPs) constitute a family of endopeptidases, which are involved in ECM and basement membranes components degradation. Fibrogenic process is characterized by altered ECM composition, associated with modifications in MMPs behavior. The active cross-talk between adipose tissue and liver can be altered in pathologies associated to insulin resistance (IR), such as NAFLD. The role of adipokines on MMPs behavior in the liver could be partly responsible of liver damage during IR. The aim of this revision is to describe the behavior of MMPs in NAFLD and its role in the associated fibrosis.

Background Nonalcoholic fatty liver disease (NAFLD) defines the clinical-pathological spectrum of hepatic lipotoxicity, which may progress to hepatic fibrosis and its complications. Thyroid hormone is a master regulator of cell metabolism and body fat distribution. Whether hypothyroidism is associated or not with an increased risk of developing NAFLD and its fibrotic progression is both clinically and physiopathologically relevant. Indeed, answering this research question would carry major pathogenic and therapeutic implications. Method PubMed database was searched using relevant key-words such as hypothyroidism; NAFLD; nonalcoholic steatohepatitis; cirrhosis; hepatocellular carcinoma; epidemiology; pathogenesis; natural history. The epidemiological studies and the meta-analyses published so far were identified as well as those studies addressing the physiopathology underlying this association. Results Many observational studies have investigated the association between either subclinical or overt hypothyroidism and NAFLD. Data are conflicting: some original and meta-analytical studies demonstrated that hypothyroidism, (mainly subclinical hypothyroidism), was common, occurring in approximately 25% of individuals with imaging-defined or biopsy-proven NAFLD; other studies, however, failed to identify a significant association between hypothyroidism and NAFLD. Moreover, such an association is biologically plausible based on the specific physiopathological impact of thyroid hormone and thyroid stimulating hormone (TSH) on metabolism of hepatocytes and accumulation and distribution of body fat. Conclusions The findings from the present review support a significant association between primary hypothyroidism and risk of development and progression of NAFLD. However, further studies evaluating the relative importance of subclinical versus overt hypothyroidism as well as addressing the mechanisms underlying the association of hypothyroidism with NAFLD are eagerly awaited.

Alcoholic and non-alcoholic fatty liver diseases are the leading causes of cirrhosis in Western countries. These chronic liver diseases share common pathological features ranging from steatosis to steatohepatitis. Fatty liver is associated with primary liver graft dysfunction, a higher incidence of complications/mortality after surgery, in correlation with impaired liver regeneration. Liver regeneration is a multistep process including a priming phase under the control of cytokines followed by a growth factor receptor activation phase leading to hepatocyte proliferation. This process ends when the initial liver mass is restored. Deficiency in epidermal growth factor receptor (EGFR) liver expression, reduced expression of Wee1 and Myt1 kinases, oxidative stress and alteration in hepatocyte macroautophagy have been identified as mechanisms involved in the defective regeneration of fatty livers. Besides the mechanisms, we will also discuss in this review various treatments that have been investigated in the reversal of the regeneration defect, for example, omega-3 fatty acids, pioglitazone, fibroblast growth factor (FGF)19-based chimeric molecule or growth hormone (GH). Since dysbiosis impedes liver regeneration, targeting microbiota could also be an interesting therapeutic approach.

Background Non-alcoholic fatty liver disease (NAFLD) is the main common cause of chronic liver disease. The aim of this study is to evaluate the effect of Shilajit, a medicine of Ayurveda, on the liver damage caused by NAFLD. Materials and methods Forty male Wistar rats, after being established as fatty liver models by feeding a high-fat diet (HFD, 12 weeks), were divided randomly into five groups as follows: control (standard diet), vehicle (HFD + distilled water), high-dose Shilajit (HFD + 250 mg/kg Shilajit), low-dose Shilajit (HFD + 150 mg/kg Shilajit) and pioglitazone (HFD + 10 mg/kg pioglitazone). The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL), glucose and liver glutathione peroxidase (GPx), superoxide dismutase (SOD) activity, malondialdehyde (MDA) levels, liver weight, and histopathological manifestation outcomes were measured after the 2-week intervention. Results Shilajit treatment significantly reduced the values of AST and ALT, TG, TC, LDL, glucose, liver weight, and steatosis, and instead, increased high-density lipoprotein (HDL) compared with the vehicle group (p < 0.05). Further, Shilajit treatment improved the adverse effects of HFD-induced histopathological changes in the liver as compared with the vehicle group (p < 0.001). MDA level and GPx activity increased but SOD activity decreased in the vehicle group compared with the control group (p < 0.05), while treatment with Shilajit restored the antioxidant/oxidant balance toward a significant increase in the antioxidant system in the Shilajit group (p < 0.05). Conclusions These findings suggest that Shilajit improved the histopathological NAFLD changes in the liver and indicated the potential applicability of Shilajit as a potent agent for NAFLD treatment.

Background 3-Iodothyronamine (T 1 AM) is an endogenous messenger chemically related to thyroid hormone. Recent results indicate significant transcriptional effects of chronic T 1 AM administration involving the protein family of sirtuins, which regulate important metabolic pathways and tumor progression. Therefore, the aim of this work was to compare the effect of exogenous T 1 AM and 3,5,3′-triiodo-L-thyronine (T3) chronic treatment on mammalian sirtuin expression in hepatocellular carcinoma cells (HepG2) and in primary rat hepatocytes at micromolar concentrations. Materials and methods Sirtuin (SIRT) activity and expression were determined using a colorimetric assay and Western blot analysis, respectively, in cells treated for 24 h with 1–20 μM T 1 AM or T3. In addition, cell viability was evaluated by the MTTtest upon 24 h of treatment with 0.1–20 μM T 1 AM or T3. Results In HepG2, T 1 AM significantly reduced SIRT 1 (20 μM) and SIRT4 (10–20 μM) protein expression, while T3 strongly decreased the expression of SIRT1 (20 μM) and SIRT2 (any tested concentration). In primary rat hepatocytes, T3 decreased SIRT2 expression and cellular nicotinamide adenine dinucleotide (NAD) concentration, while on sirtuin activity it showed opposite effects, depending on the evaluated cell fraction. The extent of MTT staining was moderately but significantly reduced by T 1 AM, particularly in HepG2 cells, whereas T3 reduced cell viability only in the tumor cell line. Conclusions T 1 AM and T3 downregulated the expression of sirtuins, mainly SIRT1, in hepatocytes, albeit in different ways. Differences in mechanisms are only observational, and further investigations are required to highlight the potential role of T 1 AM and T3 in modulating sirtuin expression and, therefore, in regulating cell cycle or tumorigenesis.

Background The global trend of obesity and diabetes is considerable. Recently, the early diagnosis and accurate prediction of type 2 diabetes mellitus (T2DM) patients have been planned to be estimated according to precise and reliable methods, artificial networks and machine learning (ML). Materials and methods In this study, an experimental data set of relevant features (adipocytokines and anthropometric levels) obtained from obese women (diabetic and non-diabetic) was analyzed. Machine learning was used to select significant features [by the separability-correlation measure (SCM) algorithm] for classification of women with the best accuracy and the results were evaluated using an artificial neural network (ANN). Results According to the experimental data analysis, a significant difference (p < 0.05) was found between fasting blood sugar (FBS), hemoglobin A 1c (HbA 1c ) and visfatin level in two groups. Moreover, significant correlations were determined between HbA 1c and FBS, homeostatic model assessment (HOMA) and insulin, total cholesterol (TC) level and body mass index (BMI) in non-diabetic women and insulin and HOMA, FBS and HbA 1c , insulin and HOMA, systolic blood pressure (SBP) and diastolic blood pressure (DBP), BMI and TC and HbA 1c and TC in the diabetic group. Furthermore, there were significant positive correlations between adipocytokines except for the resistin and leptin levels for both groups. The excellent (FBS and HbA 1c ), good (HOMA) and fair (visfatin, adiponectin and insulin) discriminators of diabetic women were determined based on specificities and sensitivities level. The more selected features in the ML method were FBS, apelin, visfatin, TC, HbA 1c and adiponectin. Conclusions Thus, the subset of features involving FBS, apelin, visfatin and HbA 1c are significant features and make the best discrimination between groups. In this study, based on statistical and ML results, the useful biomarkers for discrimination of diabetic women were FBS, HbA 1c , HOMA, insulin, visfatin, adiponectin and apelin. Eventually, we designed useful software for identification of T2DM and the healthy population to be utilized in clinical diagnosis.

Background Increasing the expression rate of recombinant mammalian hormones in Escherichia coli by combining efficient promoters and signal sequences is a never ending process. A self-induced promoter will have some beneficial gains compared to the classical T7 promoter or its variants with isopropyl β-D-1-thiogalactopyranoside (IPTG) as the inducer. Obesity is the prime suspect in widespread frequency of diabetes type II and cardiovascular diseases worldwide. YY (tyrosine-tyrosine) peptide is a local acting hormone, controlling appetite. Excitingly, it was has been shown that a truncated version of the YY peptide, YY (3-36) peptide, has potential as a worthy biopharmaceutical agent in the fight against obesity. Materials and methods To develop an economical expression system for the large scale production of the peptide in Gram-negative bacteria, we introduced a promoter sequence upstream of a chimeric gene for the extracellular expression of this peptide with the assistance of a signal sequence of asparaginase II from E. coli . This system has the advantage of producing a complete sequence of a truncated YY peptide, YY (3-36) , without any extra tags that would require further removal with the assistance of expensive specific proteases and reduced the downstream steps, significantly. Results Recombinant production of YY (3-36) peptide under a self-induced promoter proves the efficacy of the asparaginase II signal sequence as a communicator of foreign peptides and proteins into the extracellular space of E. coli . Conclusions The application of fusion protein expression of biopharmaceuticals, especially mammalian hormones, in prokaryotic systems with the help of native signal sequences makes some common tags with expensive proteases for the removal of the attached protein Tag redundant.

Background Cardiac rehabilitation (CR) is a key component of effective care for patients with a wide spectrum of cardiac problems. It seems that the short-term provision of these services can facilitate their use and meeting of related needs for patients participating in the CR programs. Materials and methods In this interventional study on patients with ischemic heart disease (IHD) referred to the Cardiac Rehabilitation Center of Chamran Hospital, Isfahan, Iran. One-hundred and twelve patients were randomly divided into two groups of 12 and 24 sessions of CR [mean age: 59.2 ± (9.02) and 60.7 ± (9.3) years, respectively] and underwent a CR program. All information was collected in two stages, before the beginning of the program and after completion of 12- and 24-session rehabilitation programs. The data collection tools were demographics questionnaire, the International Physical Activity Questionnaires (IPAQ), McNew Quality of Life Questionnaire, Beck Depression Inventory (BDI), and Zung Self-Rating Anxiety Scale (SAS). Results The results of this study showed that in both groups, total physical activity (PA) and functional capacity increased, and the quality of life (QOL) was statistically improved. Comparison of changes in mean differences at the end of the rehabilitation periods between the two groups, showed that only the weight, body mass index (BMI), waist circumference (WC) and high-density lipoprotein (HDL) in the 24-session group significantly decreased compared to the 12-session group and, but no significant difference in other variables was observed. Conclusion Overall, the findings of this study support the overall benefits of the CR program in both the short and the long term. Therefore it can be suggested that a short-term program can still be a valuable option for reducing the risk in cardiac patients and meeting their service needs.

Background To determine whether a minimal stimulation (MS) or high-dose stimulation (HDS) protocol is a better option for patients classified as poor ovarian responders (POR) in terms of reproductive and pregnancy outcomes. Materials and methods A database search for evaluation of the study outcome by using meta-analysis method was carried out. The primary outcome was the clinical pregnancy (CP) rate for each of two groups, namely, the MS and HDS groups. The secondary outcomes were the gonadotropin dose used, duration of stimulation, cancellation rate, number of oocytes retrieved, number of fertilized oocytes, number of embryos transferred and live birth rates. Results Across five databases, 4670 potential studies for further screening were selected. But ultimately only six studies, three RCTs and three retrospective or case control studies were selected that meet the Bologna criteria for POR. In all there were 624 cycles. Our meta-analysis indicated that the CP rates, cycle cancellation rates, durations of stimulation, numbers of oocytes fertilized and numbers of embryos transferred were not statistically significant. Clearly, the number of oocytes retrieved in the MS group was significantly lower than in the HDS group, while the HDS group consumed significantly higher doses of gonadotropins than the MS group. The live birth rates were significantly higher in the MS group than in the HDS group. Conclusion MS should be the first-line protocol for managing POR because the live birth rate is significantly higher, even with fewer oocytes retrieved.

Cell free DNA (cf-DNA) refers to all non -ncapsulated DNA present in the blood stream which may originate from apoptotic cells as a part of the physiological cell turnover, or from cancer cells or fetal cells. Recent studies have highlighted the utility of cfDNA analysis for genetic profiling of cancer, non-invasive prenatal testing besides many other clinical applications. In our review we discuss the sources of cfDNA in the body, the techniques most commonly being used for its isolation and analysis, the applications of cfDNA testing and the associated pros-cons. We conclude that for prenatal testing, cfDNA analysis provides an effective, non-invasive and safer alternative to traditional amniocentesis and chorionic villus sampling tests. Also, in cancer patients, cfDNA analysis is useful for genetic profiling and follow-up during treatment. However, standardization of methods of isolation and analysis has become crucial for the success of widespread use of cfDNA analysis.

The purpose of this publication is to summarize the results of the vaginal erbium:yttrium-aluminum-garnet (Er:YAG) Smooth ® laser (VEL) on the vaginal atrophy component of the genitourinary syndrome of menopause (GSM). GSM has two categories of clinical signs related to estrogen deficiency: symptoms of vulvovaginal atrophy (VVA) and urinary symptoms. This symptomatology is chronic, progressive over the years and affects a majority of women concerned by natural menopause but not exclusively: we must also consider the growing number of survivors of gynecological or non-gynecological cancers (breast, cervix, uterus, vagina, anus, etc.). At a time when hormonal treatment of menopause is contested as is the installation of under urethra prosthesis, the innovation provided by the VEL technology has the merit of offering the women concerned an effective therapeutic alternative with the security of a patent. The VEL technology has an original and unique process: acting only by thermal effect and not by ablation on tissue, VEL is a safe solution in terms of side effects and potential complications. Studies have been increasing since 2012 and all demonstrate a significant improvement in the GSM signs and symptoms, as well as an improved sexual life after VEL treatment. Double-blind, placebo-controlled, randomized studies are expected in order to ultimately confirm the safety and effectiveness of VEL.