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1.
Journal of Clinical Hepatology ; (12): 1179-1182, 2022.
Article in Chinese | WPRIM | ID: wpr-924803

ABSTRACT

Sodium taurocholate cotransporting polypeptide (NTCP) is not only an important transporter for bile acid absorption into the liver, but also a functional receptor for HBV and HDV, and extensive studies have been performed for its structure, function, gene characteristics, and expression and regulation mechanisms. NTCP is also associated with chronic viral hepatitis, nonalcoholic fatty liver disease, liver fibrosis, primary biliary cholangitis, and hepatocellular carcinoma. This article elaborates on the role of NTCP in various hepatobiliary diseases, so as to provide new direction for the diagnosis and treatment of related diseases.

2.
Journal of Clinical Hepatology ; (12): 1436-1439, 2022.
Article in Chinese | WPRIM | ID: wpr-924729

ABSTRACT

Hepatocellular carcinoma (HCC) is a type of tumor with a high incidence rate, a low rate of early diagnosis, and poor prognosis, and its development and progression involve many factors. As an important organelle in cells, mitochondria is the "energy factory" of cells and is one of the main sites for the production of reactive oxygen species in vivo, and it also participates in the regulation of cell apoptosis. There are varying degrees of changes in mitochondrial membrane, oxidation respiratory chain, mitochondrial dynamics, mitochondrial DNA, and mitochondrial calcium homeostasis during the development and progression of HCC, and such changes may affect the progression of HCC. This article systematically elaborates on the association between mitochondria and HCC, so as to provide a new direction for the diagnosis and treatment of HCC.

3.
Journal of Clinical Hepatology ; (12): 690-694, 2021.
Article in Chinese | WPRIM | ID: wpr-873818

ABSTRACT

Bile acid metabolism, gut microbiota, and bile acid receptors are involved in the development and progression of hepatocellular carcinoma (HCC). There are substantial increases in the levels of some bile acids, such as glycocholic acid, taurocholic acid, and taurochenodeoxycholic acid, in the liver tissue of HCC mice and the serum and feces of HCC patients. Bile acid metabolism due to the imbalance of the abundance of bacteria producing bile salt hydrolases and Clostridium in the intestine and the change in immune microenvironment may also promote the development of HCC. Moreover, some bile acid receptors, such as farnesoid X receptor, G protein-coupled bile acid receptor 1, pregnane X receptor, constitutive androstane receptor, and sphingosine-1-phosphate receptor 2, have been shown to participate in the development and progression of HCC through various pathways. Each link of bile acid metabolism plays a different role in the progression of HCC, and a systematic elaboration of the interaction between these links may help to deepen the understanding of the pathogenesis of HCC and develop the biological targets for early diagnosis, prognosis prediction, and precise treatment.

4.
Journal of Clinical Hepatology ; (12): 621-626, 2021.
Article in Chinese | WPRIM | ID: wpr-873808

ABSTRACT

ObjectiveTo investigate the association of high-density lipoprotein cholesterol (HDL-C) with the prognosis of patients with alcohol-related hepatocellular carcinoma (HCC) after radical treatment. MethodsA retrospective analysis was performed for the clinical data of 43 patients with alcohol-related HCC who were admitted to The Fifth Medical Center of Chinese PLA General Hospital and underwent radical treatment from January 2008 to July 2015, and according to HDL-C level, the patients were divided into normal group with 26 patients and abnormal group with 17 patients. The two groups were compared in terms of basic information, laboratory markers, imaging indices, Barcelona Clinic Liver Cancer tumor stage, and Child-Pugh class of liver function. The t-test test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The Kaplan-Meier method was used to plot survival curves and the log-rank test was used for comparison between groups. Univariate and multivariate Cox proportional hazards models were used to analyze independent risk factors for prognosis. ResultsThere was a significant difference in prealbumin between the two groups (162.38±60.86 mg/L vs 120.06±64.08 mg/L, t=2.184, P=0.035). Number of tumors (hazard ratio [HR]=2.839, 95%confidence interval [CI]: 1.120~7.200,P=0.028), tumor size (HR=2.634, 95%CI: 1.062~6.529,P=0037), and HDL-C level (HR=2.400, 95%CI: 1.040~5.537,P=0.040) were independent risk factors for the overall survival of patients with alcohol-related HCC. There were significant differences in 1-, 3-, and 5-year cumulative survival rates between the normal group and the abnormal group (88.5%/72.4%/55.7% vs 70.6%/43.7%/17.5%, χ2=5.881, P=0.015). ConclusionThe reduction in HDL-C level might indicate poor prognosis of patients with alcohol-related HCC.

5.
Journal of Clinical Hepatology ; (12): 2720-2724, 2020.
Article in Chinese | WPRIM | ID: wpr-837642

ABSTRACT

ObjectiveTo investigate the clinical effect and safety of transcatheter arterial chemoembolization (TACE) combined with microwave ablation (MWA) in the treatment of advanced primary liver cancer. MethodsA total of 186 patients with advanced primary liver cancer who were treated in The Fifth Medical Center of Chinese PLA General Hospital from April 2015 to June 2019 were enrolled and divided into study group and control group using a random number table, with 93 patients in each group. Both groups of patients underwent TACE, and the patients in the study group were treated with ultrasound-guided percutaneous MWA. The two groups were compared in terms of clinical outcome and complications. Quantitative real-time PCR was used to measure the serum level of microRNA-202 (miR-202), ELISA was used to measure the serum levels of fragile histidine triad (FHIT) and P16 protein, and the changes in the above three indices at 3 months after treatment were compared. The two-independent-samples t test was used for comparison of continuous data between two groups, and the paired t-test was used for comparison within one group before and after treatment; The chi-square testwas used for comparison of categorical data between groups. ResultsThe study group had a significantly higher objective response rate than the control group (47.32% vs 27.96%, χ2=7.422, P=0.006), and there was no significant difference in disease control rate between the two groups(P>0.05). Both groups had significant increases in the serum levels of miR-202, FHIT, and P16 protein at 3 months after treatment (all P<0.05), and compared with the control group, the study group had significantly higher serum levels of miR-202 (0.84±0.14 vs 0.58±017, t=11.385, P<0.001), FHIT (1126.35±73.05 pg/ml vs 762.87±56.71 pg/ml, t=37.904, P<0.001), and P16 protein (52.86±651 pg/ml vs 39.06±5.37 pg/ml, t=15.770, P<0.001). ConclusionUltrasound-guided MWA in addition to TACE can improve the short-term response of patients with advanced primary liver cancer and increase the serum levels of miR-202, FHIT, and P16 protein, with relatively high safety.

6.
Chinese Journal of Hepatology ; (12): 714-717, 2018.
Article in Chinese | WPRIM | ID: wpr-807391

ABSTRACT

Liver cancer is one of the most common malignant tumors in China, ranking fifth in malignant tumors and the third in tumor-related deaths. As a membrane-related protein, the asymmetric distribution of cell fate determinant Numb plays a key role in cell differentiation. Research reports that Numb may be closely associated to the occurrence and development of tumors. Recently, scholars have gradually valued its important role in liver cancer. This article briefly reviews the structure of Numb molecule, relationship between Numb and tumorigenesis, the molecular mechanism of Numb-regulated tumors, and the role of Numb in the development of liver cancer.

7.
Article in Chinese | WPRIM | ID: wpr-407744

ABSTRACT

BACKGROUND:Hepatocyte transplantation has attracted more and more attention as a therapeutic measure for liver failure and genetic metabolic liver diseases.OBJECTIVE:TO evaluate the efficacy and safety of human hepatocyte transplantation in treating hepatitis B related liver failure in one case by a 2-year follow-up.DESIGN:A case-report of 2-year follow-up.SETTING:No.9 Department of Infectious Diseases,Bioengineering Research Room,the 302 Hospital of Chinese PLA.PARTICI PANT:One inpatient with hepatitis B related liver failure was selected from the 302 Hospital of Chinese PLA.and she was diagnosed according the laboratory tests.The transplanted hepatocytes were originated frOm the healthy liver of a 24-year-old man,who had signed the protocol for liver donation before death.METHODS:The hepatocyte transplantation was completed in the Department of Radiology,the 302 Hospital of Chinese PLA in December 2004.Liver was isolated to obtain human primary hepatocytes, and then cryopreserved.The hepatocytes were transplanted into recipient spleen via femoral vein after resuscitation.The clinical symptoms,changes of blood biochemical indexes,and changes of spleen MRI signals were observed before and after operation.The patient was reexamined every half a year after operation, including liver function, blood coagulation function,B-mode ultrasonography,gastroscopy and MRI,and she was followed up for 2 years. MAIN OUTCOME MEASURES:Liver function,blood coagulation function, imaging indexes, immunological indexes,complication and rejection.RESULTS:①Totally(1-2)×1010 hepatocytes were harvested,and the viability of rewarmed hepatocytes was 60%,and finally 2×109 hepatocytes were transplanted.②Two months later,the clinical symptoms of the recipient were obviously ameliorated,and serum bilirubin and aspartate aminotransferase(AST)were obviously decreased,while prothrombin activity was markedly increased.20 months later,the MRI results showed that there was hepatocyte image in spleen.Two years after operation.the total bilirubin level was 20 μmol/L,direct bilirubin level was 7 μmol/L, alanine aminotransferase was 416.75 nkat/L,AST was 533.44 nkat/L,albumin was 37 g/L,prothrombin activity was 90%,which were all obviously ameliorated as compared with those before operation(474.5 μmol/L,340.3 μmol/L,400.08 nkat/L,1 200.24 nkat/L,38 g/L,25%).The patient left the hospital 2 months later and could do light-burdened job.No complications of hydroperitonia and liver function failure, etc.were observed,and no rejection occurred.Several reexaminations by B-mode ultrasonography all indicated the further aggravations of liver cirrhosis and esophageal varices.She was admitted to hospital for twice because of esophageal varices bleeding,and cured by endoscopic variceal sclerosis therapy.CONCLUSION:Hepatocyte transplantation can ameliorate liver function without rejection,but it cannot relieve portal hypertension.

8.
Article in Chinese | WPRIM | ID: wpr-242612

ABSTRACT

<p><b>OBJECTIVE</b>To clone the unknown gene of hepatocyte protein interacting with hepatitis C virus core protein.</p><p><b>METHODS</b>Using the yeast dual hybrid system 3, bait plasmids of hepatitis C virus core were constructed. After identifying hepatitis C virus core protein that could stably expressed in AH109 yeast strains, we performed yeast two hybrid by mating AH109 with Y187 that transformed with liver cDNA library plasmids pACT2 and then plated on quadrople dropout (QDO) medium and assayed for alpha-gal activity. The genes of yeast colonies that could grow on QDO and had alpha-gal activity were sequenced.</p><p><b>RESULTS</b>Among the 30 positive colonies, we blasted the gene of the sixth colony; we coined human hepatitis C virus binding protein 6(Hu Hcbp6) with Genbank, realized that the Hu Hcbp6 shares as much as 98% homology with two cDNA without knowing functions. We have proved that Hu Hcbp6 could interact with hepatitis C virus core protein.</p><p><b>CONCLUSIONS</b>Hepatitis C virus core binding protein (Hu Hcbp 6 Genbank number: AY032594) was successfully cloned and identified. The study partly paved the way for investigating physiological function of the Hu Hcbp6.</p>


Subject(s)
Cloning, Molecular , DNA, Complementary , Genetics , DNA-Binding Proteins , Genetics , Hepacivirus , Humans , Molecular Sequence Data , Plasmids , Sequence Analysis, DNA , Transfection , Two-Hybrid System Techniques , Viral Core Proteins , Genetics , Metabolism , Yeasts , Genetics
9.
Article in Chinese | WPRIM | ID: wpr-564621

ABSTRACT

Objective To evaluate the therapeutic effect,safety and complication of percutaneous argon-helium cryoablation in the treatment of primary hepatocellular cancer(HCC).Methods Three hundred HCC patients were treated with percutaneous argon-helium cryoablation under ultrasound guidance with argon-helium cryosurgical system.Results Two hundred and thirty-three tumors(diameter ranged from 5.0cm to 15cm,with a mean of 7.2cm?2.8cm),in 165 patients were considered to be incompletely ablated,while 185 tumors(diameter ranged from 1.9cm to 7.0cm with mean value of 5.6cm?0.8cm) in 135 patients were completely ablated.There was a significant difference in tumor diameter between these two groups(P

10.
Article in Chinese | WPRIM | ID: wpr-559636

ABSTRACT

Objective To study the changes in PAF and its receptor in Kupffer cells in experimental cirrhosis and to evaluate the role of activated Kupffer cells in portal hypertension. Methods Kupffer cells, isolated from the livers of control and CCl_4-induced cirrhotic rats, were cultured in serum-free medium overnight. PAF synthesis and release by Kupffer cells were determined 24 h later by rapid ~3H-PAF scintillation proximity assay, and the expression of PAF receptor in Kupffer cells by saturation binding technique and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). By immunohistochemisty, the distribution of PAF in Kupffer cells was surveyed. Results Cell-associated PAF synthesis and release was increased about 1.48 fold and 2 fold, respectively, by Kupffer cells in cirrhotic liver as compared with the control (P0.05). Consistent with the receptor binding capacity, the mRNA expression of PAF receptor increased significantly in the Kupffer cells of cirrhotic liver (P

11.
Article in Chinese | WPRIM | ID: wpr-554488

ABSTRACT

Objective To investigate the biological functions of hepatitis B virus e antigen (HBeAg) protein. Methods Yeast-two hybrid technique was performed to look for proteins interacting with HBeAg from hepatocytes. HBeAg bait plasmid was constructed by cloning the HBeAg gene into carrier plasmid pGBKT7, then the yeast AH109 (a type) was transformed. The transformed yeast cells was amplified and mated with yeast cells Y187 (?type) containing liver cDNA library plasmid pCAT2 in 2?YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium and were selected for two times. Results Plasmids of positive colonies were extracted and analyzed by DNA sequencing and BLAST in GenBank. Conclusions After the full sequence of new gene E-36 was amplified from the mRNA of HepG2 cells by reverse transcription polymerase chain reaction (RT-PCR) and cloned into pGADT7 vector, the recombinant plasmid was translated by reticulocyte lysate and analyzed by immuno-coprecipitation technique in vitro together with HBeAg. The result provided some new clues for the study of the biological functions of HBeAg.

12.
Article in Chinese | WPRIM | ID: wpr-553044

ABSTRACT

Interleukin 18 (IL 18) is emerging as a powerful, pleiotropic cytokine involved in determining the polarization of T cell responses. To identify the effect of IL 18 on hepatitis B virus, mouse IL 18 gene was transferred and expressed in 2.2.15 cells. Meanwhile, the inhibition of HBsAg, HBeAg and HBV DNA was investigated. Reverse transcription polymerase chain reaction (RT PCR) was used to amplify the mouse IL 18 from spleen cell of Balb/c mouse challenged with lipopolysaccharide (LPS) and phytahematoagglutinin (PHA), and reconstruct plasmid pLXSN IL18 with the retroviral vector pLXSN. Both pLXSN IL18 and pLXSN were transfected into PA317 cells and then 2.2.15 cells were infected by using the supernatant containing pseudovirus released by PA317. HBsAg and HBeAg were detected by ELISA and the HBV DNA was determined by quantitative PCR. The 580bp of mIL 18 was cloned into retroviral vector pLXSN. The pseudovirus contained in the supernatant of transfected PA317 cells was identified by RT PCR. When the pseudovirus was infected into 2.2.15 cells and incubated for 3, 5, 7, 14 days, the P/N value of HBsAg and HBeAg decreased gradually and became negative on day 14. The copies of HBV DNA reduced markedly. The results indicate that mIL 18 gene expressed in 2.2.15 cells can significantly inhibit the replication expression of HBV and may be used as a potential agent for gene therapy against HBV infection.

13.
Article in Chinese | WPRIM | ID: wpr-553815

ABSTRACT

To investigate the biological functions of hepatitis B virus e antigen (HBeAg) on pathogenesis of HBV, and to seek for effective methods to prevent and cure hepatitis B, we performed the yeast two hybrid system 3 technique to construct HBeAg bait plasmid. The bait plasmid was transformed into yeast AH109 and expressed in it. After the expression of HBeAg was identified by SDS page and Western blot, the AH109 yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2 YPDA medium to form diploid yeast and plated on synthetic dropout nutrient medium (SD/ Trp Leu His Ade) and synthetic dropout nutrient medium (SD/ Trp Leu His Ade) containing x ? gal for screening. Plasmids of blue colonies were extracted and transformed into Escherichia coli, then analyzed by DNA sequencing and bioinformatics. Thirty nine positive colonies were sequenced, among them three colonies were metallothionein. To further prove the interaction between HBeAg and metallothionein, translation was performed by using reticulocyte lysate, and immunoprecipitation was showed in vitro . The results indicated that hepatitis B virus e antigen could interact with metallothionein in vivo and in vitro.

14.
Article in Chinese | WPRIM | ID: wpr-553527

ABSTRACT

To investigate the potential role of hepatitis B virus core protein (HBcAg) in the pathogenesis of hepatitis B and seek for an effective therapeatic approach to the treatment of hepatitis B, the improved yeast two hybrid technique was employed to construct HBcAg bait plasmid, and the plasmids were transformed into yeast AH109. Then the yeast AH109 was mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium, diploid yeast was plated on both synthetic dropout nutrient medium (SD/ Trp Leu His Ade) and synthetic dropout nutrient medium (SD/ Trp Leu His Ade) containing X ? gal to be selected and screened two times. Plasmids were extracted from sixteen positive colonies, and they were sequenced. Among them, two colonies contained metallothionein. The interaction between HBcAg and metallothionein was confirmed by in vitro reticulocyte lysate translocation and immunoprecipitation. The results suggested that the pathogenesis of hepatitis B metallothionein might play an important role in the episode process.

15.
Article in Chinese | WPRIM | ID: wpr-561068

ABSTRACT

Objective To investigate the correlation between Kupffer cells and the synthesis of platelet activating factor(PAF)in experimental hepatic cirrhosis,and to elucidate the effects of endothelin(ET)on portal hypertension.Methods Thirty SD rats were randomly assigned to two groups:control group and CCl4-induced hepatic cirrhotic group.Kupffer cells,isolated from the livers of animals in both groups,were cultured for 24h.ET-1-induced PAF synthesis,and mRNA expression of PAF,ET-1 receptor and preproendothelin-1 in Kupffer cells were determined by rapid 3H-PAF scintillation proximity assay,saturation binding technique and semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR),respectively.Results Cell-associated PAF synthesis and release increased about 1.48 folds and two-folds,respectively,by cirrhotic Kupffer cells as compared to the control(1.02 ? 0.06 vs 0.69 ? 0.07 pg /mg DNA in Kupffer cells and 1.42? 0.14 vs 0.66 ? 0.04 pg/mg DNA in medium).Endothelin-1 enhanced Kupffer cells to stimulate PAF synthesis in a concentration-dependent manner,and for cirrhotic Kupffer cells,the effect was more significant than control.Cirrhotic Kupffer cells also had increased densities of functional receptors for both PAF and ET-1(exclusively ETB),but did not change the affinity of these receptors.No mRNA transcripts for the ETA receptor or preproET-1 were detected.Conclusion Kupffer cell is the main source of PAF in the cirrhotic rats.ET-1 stimulates PAF synthesis in activated Kupffer cells via ETB receptor.Since both ET-1 and PAF individually cause portal hypertension,Kupffer cells may play a role in portal hypertension associated with liver cirrhosis.

16.
Article in Chinese | WPRIM | ID: wpr-553473

ABSTRACT

To construct a subtractive cDNA library of genes transactivated by NS5A protein of hepatitis C virus with suppression subtractive hybridization technique, the mRNA was isolated from HepG2 cells transfected with pcDNA3 1(-) NS5A and pcDNA3 1(-) empty vector, respectively, then the cDNA was synthesized. After restriction enzyme RsaI digestion, small sized cDNAs were obtained. Then the tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After the tester cDNA was hybridized with the driver cDNA twice and underwent two times of nested PCR, it was then subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. The amplified library contained 121 positive clones. Colony PCR showed that 115 clones contained 200- 1 000 bp inserts. Sequence analysis was performed in 90 clones, and the full length sequences were obtained with bioinformatics method. Altogether 46 kinds of coding sequences were acquired, which consisted of 31 kinds of known and 15 kinds of unknown ones. The obtained sequences might be target genes transactivated by NS5A protein of HCV, among which some genes coding proteins involved in cell cycle regulation, cell apoptosis, signal transduction pathway and tumour development. The results indicated that the subtractive library of genes transactivated by NS5A protein of HCV was constructed successfully, which brought some new clues for studying the biological functions and pathogenesis of the viral proteins

17.
Article in Chinese | WPRIM | ID: wpr-552724

ABSTRACT

To investigate the potential role of hepatitis B virus(HBV) preS1 protein in mediating HBV adhesion to liver cell, we prepared recombinant proteins of HBV preS1 in yeast. PCR was performed to amplify the gene of HBV preS1 from the plasmid pCP10/HBV ayw subtype containing the whole fragment of HBV and the PCR product was cloned into pGEM T vector. The gene of HBV preS1 was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, the pGBKT7 plamids containing preSl were transformed into yeast cell AH109. The yeast protein was isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. The results showed that the presence of HBV presl proteins in yeast cells was confirmed by Western slot analysis. the molecular weight of the expressed product was about 30000 Da. The findings indicated that HBV preS1 was successfully expressed in yeast system.

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