Acids In Liver Tissue Research Articles

Preliminary study on the expression of MIF in HCC tissues and its relationship with ERK1/2 signaling pathway

Objective: To investigate the expression of Macrophage migration-inhibitory factors (MIF) in hepatocellular carcinoma (HCC) tissues and its interaction with ERK1/2 signaling pathway, so as to establish a theoretical basis for further studying the molecular mechanism of MIF promoting HCC. Methods: From February 2020 to August 2021, 52 cases of hepatocellular carcinoma (HCC) tissues based on hepatitis B cirrhosis (HBV-LC) and 52 cases of adjacent tissues in Tianjin Medical University Cancer Hospital and 940th Hospital of Joint Logistic Support Force of PLA were collected as the experimental group, including 39 males and 13 females, aged 35-65 years. And 20 cases of normal liver tissue were selected as the control group. Immunohistochemistry was used to detect the expression of MIF, ERK1/2 and p-ERK1/2 proteins in liver tissues of the two groups, and in situ hybridization was used to detect the expression of ERK1/2 nucleic acid in liver tissues of the two groups.HepG2 HCC cells and L-02 normal hepatocytes were co-cultured with different concentrations of rMIF, the expression and phosphorylation levels of ERK1/2 and JNK1 proteins in the two kinds of liver cells were detected by Western-blot, and the expression levels of ERK1/2 nucleic acids in the two kinds of liver cells were detected by RT-PCR. One-way ANOVA was used for measurement data and χ2 test was used for counting data. Results: The expressions of MIF, ERK1/2, p-ERK1/2 and ERK1/2 mRNA were significantly increased in HCC and para-cancer tissues (the expression of MIF in HCC group was 78.8%, and that in adjacent group was 75.0%; ERK1/2 80.8% in HCC group and ERK1/2 71.8% in paracancerous group. The expression of p-ERK1/2 75.0 % in HCC group and 46.2% in paracancerous group were respectively detected. ERK1/2 mRNA was expressed in HCC group 76.9%, ERK1/2 mRNA expression in paracancerous group 78.8%), and the differences were statistically significant compared with normal liver tissues (P<0.05), but there was no significant difference between HCC and para-cancer tissues (P>0.05). The expressions of ERK1/2, p-ERK1/2 and ERK1/2 mRNA in HepG2 HCC cells were significantly increased with the increase of rMIF concentration, and the increase was most obvious when rMIF concentration was 200 ng/ml, and the difference was statistically significant compared with L-02 normal hepatocytes (P<0.05). Conclusion: MIF, ERK1/2 and p-ERK1/2 are highly expressed in HCC tissues and HepG2 HCC cells, suggesting that MIF promotes the occurrence and development of hepatocellular carcinoma through ERK1/2 signaling pathway.

Levels and profiles of perfluorinated alkyl acids in liver tissues of birds with different habitat types and trophic levels from an urbanized coastal region of South Korea

Contamination status and characteristics of perfluorinated alkyl acids (PFAAs) including perfluorinated carboxylic acids (PFCAs) and perfluorinated sulfonic acids (PFSAs) was examined using liver tissue of birds - black-tailed gulls (Larus crassirostris), domestic pigeons (Columba livia var. domestica), pacific loons (Gavia pacifica), herons (Ardea cinerea), and egrets (Egretta garzetta and Ardea alba) - with different trophic levels, habitat types and migratory behaviors from an industrialized coastal region of South Korea. A wide range of PFAAs (1.09 ng/g to 1060 ng/g; median = 52.6 ng/g) were detected in bird livers from the Korean coasts with high detection frequency. Accumulation features of PFAAs in birds indicated that primarily trophic position and secondly habitat type influence the levels and composition of PFAAs, e.g., relatively high PFAA levels and high composition of odd-numbered long carbon chain PFCAs (perfluoroundecanoic acid (PFUnDA) and perfluorotridecanoic acid (PFTriDA)) and PFOS in higher trophic and marine birds. The prevalence of long carbon chain (≥14) PFCAs likely implies a wide use of fluorotelomer-based substances in Korea. Interspecies comparison in the accumulation profile of persistent organic pollutants (including polychlorinated biphenyls (PCBs), organochlorine pesticides, polybrominated diphenylethers (PBDEs), and PFAAs) reveals relatively high load of PFAAs in inland (pigeons) and estuarine (egrets/herons) species compared to marine bird species, indicating wide use of PFAAs in the terrestrial environment.

Occult Hepatitis B Infection in End-Stage Renal Disease Patients Starting Maintenance Hemodialysis at a Tertiary Care Hospital: A Descriptive Cross-sectional Study.

Occult hepatitis B infection is defined as the presence of the hepatitis B virus deoxyribonucleic acid in liver tissues and/or serum in the absence of serum hepatitis B Virus surface antigen. The prevalence of occult hepatitis B infection in end-stage renal disease patients is largely unknown. The aim of the study is to determine the prevalence of occult hepatitis B infection in the hemodialysis population starting maintenance hemodialysis. A descriptive cross-sectional study was conducted in the department of Internal Medicine of a tertiary care hospital. Convenience sampling method was used; 50 consecutive end-stage renal disease patients, who started maintenance hemodialysis from March 2019 to March 2020, were enrolled in the study. The study was approved by the Institutional Review Committee of the hospital (reference number: 351/2019). Statistical Package for Social Sciences version 26.0 was used for statistical analysis. The mean age of the patients was 50.34±12.65 years, and 42 (84%) were male. About 4 (8%) patients were diagnosed having occult hepatitis B infection, 3 (6%) of them were seropositive and 1 (2%) seronegative. About 41 (82%) patients had no history of hepatitis B vaccination series before starting hemodialysis; 36 (72%) had anti-hepatitis B surface antibody titre <10 mIU/ml. About 44 (88%) patients received a blood transfusion during their hemodialysis sessions and 14 (28%) patients had a history of receiving hemodialysis at other centres. Our study demonstrated a high prevalence of occult hepatitis B infection among end-stage renal disease patients starting hemodialysis.

Tyrosine metabolic enzyme HPD is decreased and predicts unfavorable outcomes in hepatocellular carcinoma

BackgroundLiver is a major metabolic organ containing many metabolic enzymes. Disorders of liver-specific enzymes can cause liver dysfunction and tumorigenesis. Previous studies indicated that 4-Hydroxyphenylpyruvate dioxygenase (HPD) plays an essential role in catalyzing the tyrosinolytic metabolism of 4-hydroxyphenylpyruvate to homogeneous acids in liver tissues. However, the clinical significance of HPD in HCC has not been obtained. Here in our study, we aimed to identify the expression and the clinical significance of HPD in hepatocellular carcinoma (HCC). MethodsWestern Blotting and qRT-PCR were employed to evaluate the level of HPD in HCC cell lines and fresh samples. The expression of HPD was further confirmed by immunohistochemistry (IHC) using a tissue microarray (TMA) cohort with a total of 778 HCC patients. Furthermore, the mRNA expression of HPD in HCC was evaluated from TCGA and GEO public databases. Kaplan-Meier analysis and univariate and multivariate Cox regression analyses were used to determine the correlation between HPD expression with clinicopathological variables and survival rate of HCC patients. The cellular behaviors of transfected cells were respectively examined by CCK8 and Migration assay. ResultsThe expression of HPD is restricted in liver compared with other cancer types. HPD mRNA and protein expression was dramatically reduced in HCC cell lines and fresh tissue samples. IHC staining in HCC TMA further showed that the decreased of HPD in paraffin-imbedded HCC samples was linked to an adverse overall postoperative survival (p < 0.001). Clinicopathologically, low expression of HPD was correlated with larger tumor size, advanced TNM staging and poor differentiaion. In addition, multivariate analyses indicated that HPD was an independent predictive factor of HCC survival. Our study pioneering validates that knockdown of HPD increases HCC cell cell growth and cell motility. ConclusionOur results suggested that HPD may serve as a valuable prognostic marker, a tumor suppressor, and a potential therapeutic target for HCC patients.

Cafeteria diet from birth to adulthood promotes hepatic steatosis and redox imbalance in Wistar rats

Purpose The purpose of this paper is to evaluate the pathological, biochemical and redox state parameters of liver tissue in Wistar rats treated from birth to adulthood (119 days) with cafeteria diet. Design/methodology/approach During the lactation, 6 liters of Wistar rats (dam + 8 pups each) were fed one of two diets: control (CTRL; n = 3) or cafeteria (CAF; n = 3) diets and water ad libitum. After weaning, the males were placed in individual cages, receiving the same diet offered to their respective dams (CTRL or CAF; n = 18) until adulthood. The following parameters were evaluated: absolute and relative liver weight; blood, liver and feces biochemistry; liver histology; and redox state of the liver. Findings When assessing the relative and absolute organ weight, no significant differences were found between the groups. The Cafeteria group exhibited higher values of serum LDL-c (p = 0.008), VLDL-c (p = 0.03) and triglycerides (p = 0.01), as well as several micro and macrovacuoles of fat accumulation, higher hepatic lipid (p = 0.03) and cholesterol (p = 0.0001) levels regarding Control group. Cafeteria group showed greater expression of glutathione-s-transferase (p = 0.03) and superoxide dismutase (p = 0.005) enzymes compared to the control group. In the case of the markers of oxidative stress, there was no difference between the groups. Originality/value A simple and standardized cafeteria diet caused an accumulation of fatty acids in liver tissue, inducing a state of hepatic steatosis besides an increased expression of antioxidant enzymes.

First report of equine parvovirus-hepatitis-associated Theiler's disease in Europe.

Equine parvovirus-hepatitis (EqPV-H) has been proposed as the aetiological cause of Theiler's disease, also known as serum hepatitis. EqPV-H-associated Theiler's disease has not been previously reported in Europe. To determine whether EqPV-H infection was associated with a 2018-2019 outbreak of Theiler's disease in four horses on a studfarm. Descriptive case series. The medical records of four horses from the same farm diagnosed with fatal Theiler's disease were examined retrospectively. Information collected included a clinical history, physical examination findings, tetanus antitoxin exposure, serum biochemistry and necropsy reports. Liver tissue from all four horses was tested for EqPV-H using PCR and in situ hybridisation (ISH) assays. Three of the horses had a history of recent (7-11weeks) tetanus antitoxin administration. Liver tissue from all four horses tested positive for EqPV-H with PCR. In situ hybridisation revealed a widespread distribution of viral nucleic acid in hepatocytes in one case, and a more sporadic distribution in the remaining three cases. Case controls were not available from the farm in question given the retrospective nature of analysis. This case series documents the first reported EqPV-H-associated Theiler's disease in Europe and the first use of ISH to visualise the viral nucleic acid in liver tissues of horses with Theiler's disease.

Dietary supplementation with L-arginine and combinations of different oil sources beneficially regulates body fat deposition, lipogenic gene expression, growth performance and carcass yield in broiler chickens

Context Broiler meat with excessive of fat and saturated fatty acids content has serious health implication for consumers. The accumulation of abdominal fats in broiler chickens constitutes a loss of dietary energy and also reduces carcass yield. Oil rich in unsaturated fatty acids and l-arginine are effective for reducing fat deposition and improve meat quality. Aims The aim of this study was to examine the effects of supplementation of l-arginine (l-Arg) with four combinations of palm oil (PO) and sunflower oil (SO) on growth performance, carcass yield, fat deposition, lipogenic gene expression and blood lipid profile in broiler chickens. Methods A total of 180 1-day-old chicks (Cobb 500) were randomly assigned to five dietary treatments as: T1, 6% PO (control); T2, 6% PO + 0.25% l-Arg; T3, 4% PO + 2% SO + 0.25% l-Arg; T4, 2% PO + 4% SO + 0.25% l-Arg; and T5, 6% SO + 0.25% l-Arg. Key results Birds fed l-Arg and combinations of PO and SO had higher weight gain at starter and finisher period compared with the control. The carcass yield increased, and relative abdominal fat reduced in broiler fed with combinations of l-Arg and increased level of SO in the diet. The concentration of oleic, palmitoleic and total monounsaturated fatty acids in liver tissue decreased by addition of l-Arg in broiler diet. The palmitic and total saturated fatty acid decreased, and total unsaturated fatty acid and polyunsaturated fatty acids increased in liver tissue when PO replaced progressively by SO supplemented with l-Arg in the diet. The acetyl-CoA carboxylase , stearoyl-CoA desaturase and fatty acid synthetase gene expression tended to decrease by supplementation of l-Arg with an increased level of SO compared with control. Conclusion Supplementation with l-Arg and combination of PO and SO at the ratio of 4:2 could inhibit lipogenesis and subsequent lower abdominal fat deposition and enhance growth performance and carcass yield in broiler chickens. Implications Ratio of PO and SO, 4:2 with l-Arg supplementation in the dietary of broiler chickens can contribute to a better growth performance, lesser fat deposition and greater carcass yield.

Co-inertia multivariate approach for the evaluation of anthropogenic impact on two commercial fish along Tyrrhenian coasts

Aliphatic hydrocarbon levels were determined by the GC/MS technique in fish livers of Engraulis encrasicolus (Ee) and Trachurus trachurus (Tt), collected from a particular area of the Mediterranean Sea, called GSA 10, which is located exactly in Tyrrhenian Sea between Campania coast and North Sicily coast. The aim was to evaluate their potential use as specific bioindicators towards this class of contaminants. Both Tt and Ee are considered to be pollution monitoring bioindicators, due to their dominance in marine communities and economic fishing interest. Ee showed a higher tendency to bioaccumulate TAHs, due to the lower quantity of fatty acids in liver tissues with respect to Tt. The area under study has been characterised a) chemically with the acquisition of temperature, oxygen and salinity profiles along the water column, and b) ecologically with the determination of amino acid contents in fish eyes, in order to gain information on the adaptation to environmental changes. Moreover, specific activities of two hydrolytic enzymes, such as alkaline phosphatase and peroxidase in fish epidermal mucus, together with lactate in blood plasma and cortisol levels, have been investigated for the first time, in order to obtain insights into the effects of hydrocarbons on animal welfare. A multiple co-inertia analysis was also applied to chemical and environmental parameters, in order to explore any possible correlation between different variables. The multivariate approach showed a clear spatial distribution between environmental and chemical variables in Ee, whilst there was an absence of a spatial trend in Tt. Moreover, the chemometric analysis showed a very high correlation between amino acid profiles and environmental variables for both species, confirming the possibility of being used as ecological welfare indices for short-term environmental variations.

Combinatorial treatment of Rhizoma Paridis saponins and sorafenib overcomes the intolerance of sorafenib

Sorafenib, as a multi-kinase inhibitor, was the first FDA-approved anti- hepatocellular carcinoma (HCC) drug. Rhizoma Paridis saponins (RPS) as natural products have shown antitumor activity through regulation of glycolytic and lipid metabolism which was regarded as the side effect limited the utility of sorafenib. In this research, we tried to use metabolomics to verify the probability of combinatorial treatment of RPS and Sorafenib. As a result, Sorafenib + RPS increased the antitumor effect of sorafenib and RPS in H22 mice. They mitigated the change of liver weight and the increasing levels of AST and ALT in serum, and AFP and MDA in liver tissues, which indicated their liver protective activity. They also up-regulated the activity of NOX and SDH, concentration of ATP, and down-regulated the mRNA and protein levels of HIF-1a and concentration of lactate, which suggested they protected against mitochondria damage and inhibited anaerobic glycolysis. Meanwhile, the combination group remarkably down-regulated the concentration of octadecanoic acid and hexadecanoic acid in serum, and tetradecanoic acid in liver tissues compared with model group (p < 0.05). Relative regulation mechanism included their decreasing mRNA levels of FASN, CPT1, GLUT1, Myc, Akt, mTOR and LDHA, and increasing the protein expression of p53 in tumor and liver tissues (p < 0.05). Furthermore, similar influence can be observed in protein levels of CPT1A, p-PI3K, p-mTOR and p53 in liver tissues and FASN in serum. All of that provided possibility to overcome the intolerance of sorafenib by drug compatibility through protection against mitochondria damage, inhibition of anaerobic glycolysis and suppression of lipid synthesis based on PI3K/Akt/mTOR pathway.

Identification of glycerophospholipid fatty acid remodeling by using mass spectrometry imaging in bisphenol S induced mouse liver

Bisphenol A (BPA) plays an important role in metabolic disorders. As a major alternative to BPA, it is unclear whether the exposure of bisphenol S (BPS) may result in lipidome disturbance. Using a mouse model, we investigated the effects of BPS exposure on metabolism and spatial distribution of lipids by using lipidomics analysis and matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry imaging (MSI) in mouse liver tissues. Lipid metabolites displayed significant up-regulation in phosphatidylethanolamines (PE), lysophosphatidylcholines (LPC), lysophosphatidylethanolamines (LPE) and lysophosphatidylserine (LPS) as well as remarkable down-regulation in phosphatidylcholine (PC) and phosphatidylserine (PS) in mouse liver after the exposure at 100μg BPS/kg body weight/day. The obtained results indicated that the lipidome of liver was perturbed significantly in glycerophospholipid (GP) fatty acid remodeling pathway upon the BPS exposure. We applied MSI and multivariate statistical analysis to evaluate the abundance variation of lipid markers in BPS-treated liver sections and to compare with the analytical results from olive oil-treated liver sections. Differential structural lipids with up-regulated PE (20:1/20:4), LPC (20:4), LPE (20:4), LPS (33:4) and down-regulated PC (20:4/22:6) and PS (18:0/22:6), which were related to GP fatty acid remodeling, changed and co-localized in the liver sections. To explore the cause of variation of lipid abundance, expression of enzymes that regulate biosynthesis and metabolism of fatty acid in liver tissues were analyzed. Consistent with the results of liver lipidome and spatial distribution, a decrease in hepatic expression of LPC acyltransferase 1 (LPCAT1), LPCAT2 and LPS acyltransferase and an increase expression of LPCAT3, LPCAT4, LPE acyltransferase 1 (LPEAT1), LPEAT2 and phospholipase A2s were observed in GP fatty acid remodeling pathway. Our results demonstrated that exposure to BPS could induce the GP fatty acid remodeling, which might be useful in toxicity evaluation for bisphenols-induced hepatic diseases.

Expression and Association of SCD Gene Polymorphisms and Fatty Acid Compositions in Chicken Cross

Stearoyl-CoA desaturase (SCD) is an integral membrane protein of endoplasmic reticulum (ER) that catalyzes the rate limiting step in the monounsaturated fatty acids from saturated fatty acids. Selection for fatty acids traits based on molecular marker assisted selection is needed to increase a value of chicken meat. This study was designed to analyze expression and associations of SCD gene polymorphisms with fatty acid traits in F2 kampung-broiler chicken cross. A total of 62 F2 kampung-broiler chicken cross (29 males and 33 females) were used in this study. Fatty acid traits were measured at 26 weeks of age. Samples were divided into two groups based on fatty acid traits (the highest and the lowest). Primers in exon 2 region were designed from the genomic chicken sequence. The SNP g.37284A>G was detected and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was then used to genotype. The expression of SCD gene was analyzed using quantitative real time PCR (qRT-PCR). The result showed that there were three genotypes (AA, AG, and GG) found in this study. The SCD|AciI polymorphism was significantly associated with palmitoleic acid (C16:1), fatty acids total and saturated fatty acid in 26 weeks old of F2 kampung-broiler chicken cross (P<0.05). The SCD gene was expressed for polyunsaturated fatty acids in liver tissue in two groups of chickens. In conclusion, the SCD gene could be a candidate gene that affects fatty acids traits in F2 kampung-broiler chicken cross.

Umbelliferone Improves an Impaired Glucose and Lipid Metabolism in High-Fat Diet/Streptozotocin-Induced Type 2 Diabetic Rats

Umbelliferone (UMB) is a natural product that has several pharmacological effects including antihyperglycemic activity in diabetic rats. Thus, the objective of this study was to investigate the effect of UMB on insulin resistance and on the regulation of glucose and lipid metabolism in type 2 diabetic rats. Type 2 diabetes was induced in rats by feeding a high-fat diet (45 kcal% fat) and a single dose of streptozotocin injection. After 8 weeks of treatment, UMB significantly reduced the elevated blood glucose levels and insulin resistance and increased the liver glycogen and serum adiponectin. Moreover, the serum lipid and the storages of triglyceride and non-esterified fatty acid in liver tissue were reduced. From histological examination, the lipid droplets in liver tissue were clearly decreased, and the fat cell size in the fat tissue was smaller in diabetic rats treated with UMB. Interestingly, UMB increased fat cell adiponectin, plasma membrane glucose transporter 4 (GLUT4) and peroxisome proliferator-activated receptor gamma (PPARγ), and liver PPARα protein expressions. Our findings demonstrate that UMB improves glucose and lipid metabolism in type 2 diabetes by stimulating the insulin secretion and the related mechanisms via stimulating expression of adiponectin, GLUT4, PPARγ, and PPARα-protein expressions. Copyright © 2015 John Wiley & Sons, Ltd.

Superoxid dismutase activity in portal vein endothelium after partial liver resection

To investigate superoxide dismutase (SOD) activity in the portal vein endothelium and malondialdehyde acid (MDA) production in liver tissue of rats submitted to 70% hepatectomy. Twelve rats were distributed in two groups (hepatectomy and sham). Animals were sacrificed on post operative day 1 and portal vein, liver tissue and blood samples were collected. Portal vein SOD production was measured using lucigenin-amplified chemiluminescence assays. MDA measurement was used as an index of oxidative stress through the formation of TBARS (Thiobarbituric Acid Reactive Species). There was no difference in post operative bilirubin, AST, ALT levels between groups. DHL level was higher in the hepatectomy group (p=0.01). MDA production in the remnant liver tissue and endothelial portal vein SOD activity were also significantly (p<0.05) elevated in the hepatectomy group when compared to control group. There was no correlation between MDA and SOD activity. SOD activity, on the other hand, showed a positive correlation with LDH level (p=0.038) and MDA levels showed a positive correlation with AST and ALT levels (p<0.001). There is an increased production of malondialdehyde acid in liver tissue after partial hepatectomy and increased activity of superoxide dismutase in portal vein endothelium as well.

Increased Protein Oxidation and Loss of Protein-Bound Sialic Acid in Hepatic Tissues of D-galactose Induced Aged Rats

A redox basis of the increased oxidative protein damage and free radical-mediated desialylation have not been fully elucidated in aging. It is well known that the incidence of several liver diseases increase with age. This original research focuses on protein oxidation mechanisms and protein-bound sialic acid levels in liver tissue of the mimetic aging rats. Injection of D-galactose (60 mg/kg/day) for six weeks to male Sprague-Dawley rats (20-week-old) used to establish mimetic aging model. We investigated the tissue levels of various protein oxidation markers such as protein carbonyl groups, suitable advanced oxidation protein products and protein thiol groups. Our study also covered protein-bound sialic acid in liver tissue of D-galactose-induced aging rats. PCO (Protein Carbonyl Groups), P-OOH (Protein Hydroperoxides) and AOPP (Advanced Oxidation Protein Products) levels in aging rats were significantly higher compared to young control groups. On the other hand, P-SH (Protein Thiol Groups) levels were not found to be different between two groups. SA (Sialic Acid) levels in D-galactose-induced aging rats were significantly lower compared to control groups. Our results demonstrated greater susceptibility to hepatic oxidative protein damage and desialylation of hepatocellular proteins in Dgalactose- induced aging rats. These molecular mechanisms may be operative in the many age-related liver diseases, which are pertinent to increased oxidative stress and altered redox homeostasis.

Therapeutic effect of coenzyme Q10 against experimentally-induced hepatocellular carcinoma in rats

The therapeutic potential of coenzyme Q10 was investigated in rats with hepatocellular carcinoma induced by trichloroacetic acid (0.5g/kg/day, p.o., for five days). Coenzyme Q10 treatment (0.4mg/kg/day, i.p.) was applied for four weeks following trichloroacetic acid administration. Coenzyme Q10 significantly suppressed lipid peroxidation, prevented the depletion of reduced glutathione and superoxide dismutase activity, and decreased the elevations of tumor necrosis factor-α and nitric oxide in liver tissue of rats with hepatocellular carcinoma. Also, the histopathological dysplastic changes induced by trichloroacetic acid in liver tissue were ameliorated by coenzyme Q10. Immunohistochemical analysis revealed that coenzyme Q10 significantly decreased the expression of hepPar-1, alpha-fetoprotein, inducible nitric oxide synthase, cyclooxygenase-2 and nuclear factor-κB in liver tissue of rats with hepatocellular carcinoma. It was concluded that coenzyme Q10 may represent a potential therapeutic option for liver carcinogenesis.

Novel Approach for Quantification of Hepatitis C Virus in Liver Cirrhosis Using Real-Time Reverse Transcriptase PCR

BackgroundHepatitis C virus (HCV) infects nearly 3% of the population worldwide and is a major cause of acute and chronic infections leading to fibrosis, cirrhosis, and hepatocellular carcinoma. Current laboratory diagnosis of HCV is based on specific antibody detection (anti-hepatitis C virus (anti-HCV)) in serum. As HCV replicates in the liver cells, detection and localization of HCV RNA in liver tissue are vital for diagnosis. MethodsTen biopsy samples diagnosed for cryptogenic liver cirrhosis, negative for the presence of anti-HCV and serum HCV RNA, were studied for analyzing presence of viral nucleic acid in liver tissues. Qualitative screening for HCV was done through ELISA while the nucleic acid analysis was performed through COBAS Amplicor. Detection of HCV RNA in liver tissue biopsies was performed following standard protocol of HCV detection kit (Shenzhen PG Biotech) with modifications using Light Cycler 2.0 (minimum detection limit 10 copies/ml). ResultQuantitative detection in liver biopsies following the modified method showed the presence of HCV RNA in three samples out of the ten studied. ConclusionThe results indicate that using Light Cycler 2.0, following the modified technique described, constitutes a reliable method of quantitative detection and localization of HCV in tissue in “serosilent” HCV infection.

Chemoprotective Effect of Sobatum against Lithium-Induced Oxidative Damage in Rats

Lithium therapy mainly used in curing some psychiatric diseases responsible for numerous undesirable side effects on different organs in humans. The present study explores the beneficial effect of sobatum, a purified compound of Solanum trilobatum, on lithium carbonate (Li2CO3)-induced multiple organ toxicity in rats. Li2CO3 (150 mg/kg body weight) was administered orally in drinking water for a period of 30 days to induce toxicity in rats. Li2CO3 could induce lipid peroxidation to a significant extent that was accompanied by marked reduction in reduced glutathione, SOD, CAT, GST, GPX activities, and parallel decline in ATP in tissues. Toxicity resulted in abnormal elevation of lipids such as cholesterol, triglycerides, phospholipids, and fatty acids in liver tissues. Treatment with sobatum affords substantial protection in liver and heart by altering all the parameters to near normal levels that were further confirmed by histological examination. Sobatum prevents Li2CO3-induced oxidative damage of DNA by reducing DNA fragmentation indicating its block on cell death. However, these results demonstrated that sobatum has the ability to suppress the drug-induced toxicity.

Gas Chromatography-Mass Spectrometry Based on Metabonomics Study of Carbon Tetrachloride-Induced Acute Liver Injury in Mice

A gas chromatography-mass spectrometry based on methodology was developed to investigate the distinctions of metabolic profiles in liver tissues and plasma of mice with different time post-dose after CCl 4 treatment. NIST mass spectral library was used for the rapid identification of endogenous metabolites, and the principal component analysis was introduced for the pattern recognition of mice metabonome. The pattern recognition result presented clear changing tendencies of metabolite profiles at various time postdose, both in liver tissues and plasma. The elevated levels of malate and several fatty acids in liver tissues as well as the increased concentrations of some amino acids and citrate in plasma were detected as metabolic effects of CCl 4 induced liver injury. Changing tendencies of the metabolite profiles and several metabolites were conformed to the histopathological assessment of the liver tissues. The results suggested that the GC-MS based method can provide some advantages in metabolite profiling and compound identification, and therefore can be employed as a potential powerful approach in metabonomics research as a complement of NMR.

Comparison of the metabolism of ethylene glycol and glycolic acid in vitro by precision-cut tissue slices from female rat, rabbit and human liver

1. The metabolism of [1,2-14C]-ethylene glycol and [1,2-14C]-glycolic acid was studied in vitro using precision-cut tissue slices prepared from the livers of female Sprague—Dawley rats, New Zealand white rabbits and humans. The time-course for production of metabolites formed from ethylene glycol at concentrations from 3 to 40 mM was determined to compare quantitatively the differences between species in the rates and amounts of formation of glycolic acid, the presumed developmental toxicant of ethylene glycol. The rates of metabolism of glycolic acid to glyoxylic acid at concentrations from 0.05 to 16 mM by liver tissue from the different species were also determined. The apparent VmaJKm for the metabolic conversions of ethylene glycol to glycolic acid and for glycolic acid to glyoxylic acid in liver tissue from the different species were obtained.2. There were qualitative differences in the metabolic profiles and quantitative differences in the formation of glycolic acid between the mammalian liver systems. There was an average of 10-fold less glycolic acid produced by liver slices from rabbits compared with rats. With the human liver, the formation of glycolic acid was not detectable using tissue from three of four human donors. A low level of glycolic acid was detected in one liver slice incubation from one of the four subjects, but only at one extended time point; glyoxylate was detected with liver slices from all four humans.3. Liver slices prepared from female Sprague—Dawley rats, female New Zealand White rabbits and three female human subjects all metabolized glycolic acid to glyoxylic acid. Human liver tissue was the most effective at further metabolizing glycolic acid to glyoxylic acid. The ratios of F"max/-/Cm, representing the relative clearance of glycolic acid from liver tissue, were approximately 14:9:1 for human, rat and rabbit liver, respectively.4. Precision-cut liver slices maintained in dynamic organ culture are good predictors of metabolism by liver tissue in vivo. The results of the present study therefore indicate that levels of glycolic acid, if formed in vivo, following exposures to similar concentrationsof ethylene glycol, would be lower in humans than in rabbits and rats.

Hepatitis B virus nucleic acids circulating in the blood: distinct patterns in HBs carriers with hepatocellular carcinoma.

Circulating nucleic acids in serum and plasma are common in a variety of disease conditions. Here, we focus (i) on our approach for the detection of various hepatitis B virus (HBV)-related nucleic acids in liver tissue and in serum, (ii) on the progression of the chronic HBV infection, (iii) on the relation of HBV-specific nucleic acids circulating in the blood of patients with hepatocellular carcinoma, and in general (iv) on the diagnostic potential of circulating HBV nucleic acids.