Clinical application of serum Golgi protein 73 in patients with chronic liver diseases / 中华肝脏病杂志

Golgi protein 73 (GP73) is a transmembrane protein on the Golgi apparatus and can be cut and released into the blood. In recent years, an increasing number of clinical studies have shown that the elevated serum GP73 level is closely related to liver diseases. And thus GP73 is expected to be used as a new serum marker for assessing progress of chronic liver diseases. Herein, the clinical application of serum GP73 in chronic hepatitis, liver fibrosis, liver cirrhosis and hepatocellular carcinoma with different etiologies was reviewed based on available literatures; and a research outlook in this field is made.

Thirty years' achievements in prevention and control of hepatitis B virus infection / 中国实用内科杂志

Hepatitis B vaccination and blocking mother-to-child transmission of HBV are of great significance in the prevention and control of HBV infection. Hepatitis B vaccination is the most effective way to prevent HBV infection. Mother-tochild transmission of HBV can be greatly reduced by these measures, which include strengthening the screening of HBV in women of childbearing age, antiviral treatment in pregnant women with high viral load, and combined immunization of hepatitis B vaccine and hepatitis B immunoglobulin for newborns of HBsAg positive mothers. In the recent 30 years, remarkable achievements have been made in the prevention and control of HBV infection in China.

Clinical characteristics of drug-induced autoimmune hepatitis / 中国实用内科杂志

OBJECTIVE: To investigate the clinical characteristics of drug-induced autoimmune hepatitis(DIAIH), in order to help clinicians learn more about DIAIH and improve its clinical diagnosis. METHODS: This was a retrospective study in the patients with DIAIH and DILI treated at Shengjing Hospital of China Medical University between January 2013 and December 2017. The population characteristic,related drugs,clinical manifestations,liver biochemical parameters, autoimmune antibodies, and liver pathological features were compared between the two groups. RESULTS: There were 49 patiens with DIAIH and 436 patiens with DILI. The majority of these patients were female. There was a significant difference in the proportion of female patients, with DIAIH(91.84%, 45/49)higher than DILI(70.41%, 307/436)(χ~2=9.111, P=0.003). The patients with DIAIH had a mean age of(50.0±7.4) years. The top three drugs inducing DIAIH were Chinese herbal medicines(53.06%, 26/49), non-steroidal anti-inflammatory drugs(10.20%, 5/49) and fluvastatin(10.20%, 5/49). DIAIH occurred after more than 8 weeks of treatment. There was a significant difference in the proportion of liver failure, with DIAIH(30.61%, 15/49) higher than DILI(14.68%, 64/436)(χ~2=8.20, P=0.004). The risks of DIAIH and DILI with liver failure caused by western medicines were significantly higher than those by Chinese herbal medicines(χ~2=9.77, P=0.002; χ~2=16.09,P<0.001). The positive rate of autoimmune antibody in DIAIH patients was 100%. The positive rate of ANA was 83.67%(41/49), and that of SMA was 16.33%(8/49). The liver pathological features of interface hepatitis, plasma cells and lymphocytes infiltration in DIAIH patients were more obvious than those in DILI patients. CONCLUSION: DIAIH is more common in the female patients and is caused frequently by Chinese herbal medicines. DIAIH caused by western medicines could easily result in liver failure.

Clinical difficulties and hotspots of liver injury caused by non-hepatitis virus / 中国实用内科杂志

There are many viruses causing liver injury in clinic, including hepatophilic and non-hepatophilic viruses. With the extensive vaccination of hepatitis B vaccine and strict management of blood products in newborns, patients with chronic hepatitis B and C will gradually decrease, while liver damage caused by non-hepatitis virus will gradually increase, which should be paid much attention to highly valued by clinicians.

Comparative study on etiology and complications of liver cirrhosis in a 5-year period / 中国实用内科杂志

OBJECTIVE: To investigate the variation of etiology and complication of liver cirrhosis(LC) by the comparative analysis of etiology,complications, sex and age in LC patients in 2012 and in 2017. METHODS: In this cross-sectional study, we collected cases of LC admitted in 2012 and 2017 and reviewed the medical records. The demographics, etiology and complications were collected and we compared the composition ratios of etiology and complications as well as the sex composition and age differences between different etiology in the 5-year period. RESULTS: 3065 patients(including 1451 in 2012 and 1614 in 2017) were identified in this study. There was no significant difference in etiology of LC caused by HBV infection(that was 56.31% in 2012 and 53.41% in 2017, respectively.(χ2=2.591, P=0.107). The composition ratio of alcohol and autoimmune diseases increased. That of alcohol diseases was 12.96% in 2012 and 16.36% in 2017(χ2=7.027, P=0.008).That of autoimmune diseases was 9.92% in 2012 and 13.07% in 2017(χ2=7.398, P=0.007). The composition ratio of HCV infection decreased from 14.82% to 11.28% having statistically significant difference(χ2=8.497, P=0.004). The three former complications in 2012 were UGH(15.64%), HCC(15.30%,), SBP(12.68%,), which were HCC(21.07%), UGH(13.38%), SBP(11.03%) in 2017. HCC was more common(that was 15.30% in 2012 and 21.07% in 2017) having significant difference(χ2=16.964, P<0.001).LC caused by HBV and alcohol were mainly males, which slightly decreased having no significant difference. LC caused by autoimmune diseases was mainly female, which slightly increased having no significant difference. The LC patients infected by HBV and HCV were older than before when were hospitalized.That of HBV was(50.08±11.11) years old in 2012 and(52.39±11.56) years old in 2017(t=-4.163, P=0.004). That of HCV was(57.22±10.52)years old in 2012 and(61.13±10.25) years old in 2017(t=-3.732, P <0.001). CONCLUSION: Compared with 5 years ago, HBV infection remained the major cause of liver cirrhosis, whereas alcohol and autoimmune diseases increased and HCV infection decreased. HCC was the most common of LC complications. LC patients caused by different etiology had different prevalence in sex and were hospitalized in different ages. Patients infected by HBV/HCV seemed to be older than before when they were hospitalized.

Quantitative analysis of HBV cccDNA from liver biopsy specimens in HBV carriers: correlation with serological makers and sera HBV DNA / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the correlation of sera HBV DNA and serological makers with hepatic tissue HBVcccDNA in chronic HBV carriers.</p><p><b>METHODS</b>Real time fluorescence quantitative polymerase chain reaction (RT-PCR) were used to detect HBV covalently closed circular DNA (cccDNA) and total intrahepatic HBV DNA from 30 needle-biopsy specimens as well as HBV DNA in sera in chronic HBV carriers. Quantification of the HBsAg, HBeAg in sera were quantified using Chemiluminescence immunoassay.</p><p><b>RESULTS</b>HBVcccDNA can be detected in chronic HBV carriers, which rang from 3.15 x 10(3) copies/mg to 1.06 x 10(7) copies/mg. There was a positive correlation between the cccDNA and HBVtDNA (r = 0.375, P < 0.05), but there was no correlation between the cccDNA and sera HBV DNA (P = 0.174). There was a positive correlation between cccDNA and sera HBsAg quantification (r = 0.562, P < 0.001) but no correlation with sera HBeAg qantification (r = 0.152, P > 0.05).</p><p><b>CONCLUSION</b>HBV cccDNA can be replicated stably in hepatic tissue in all chronic HBV carriers. HBV DNA in sera can not be indicated hepatic tissue cccDNA level. While HBsAg quantification in sera can be used as a marker of cccDNA quantification in hepatic tissue to some extent.</p>

Study on the relationship between the serum levels of Th1/Th2 cytokines and the clinical manifestations of chronic hepatitis C and the outcome of interferon therapy / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigated the relationship between the serum levels of Th1/Th2 cytokines and the progress of viral hepatitis C and the outcome of interferon therapy.</p><p><b>METHODS</b>Serum cytokine detection used the method of ELISA. HCV genotype were classified by direct sequencing. HCV RNA loads were determined by fluorescence quantitative PCR.</p><p><b>RESULTS</b>The levels of IL-2 and TGF-beta in serum of patients with chronic hepatitis C were lower hut IL-5 was higher than those of normal control. The level of IL-6 was positively related to the sera level of ALT and was negatively related to sera HCV RNA load. Patients of HCV genotype 1 had higher sera quantities of IL-6 than those of genotype 2 and patients of genotype 2a had lower sera quantities of IL-2 than those of 2b. The levels of IL-2 had the tendency to decrease whereas IL-6 had the tendency to increase when time went on. The level of TGF-beta increased at early phase but decrease later. There were no difference of all cytokines detected between the groups of response and nonresponse before interferon therapy, hut the quantity of serum IFN-gamma were increased after interferon therapy in the response group.</p><p><b>CONCLUSION</b>The tested cytokines co-participate in the pathogenesis of chronic hepatitis C and have the relationship with the clinical manifestations of the patients. There were no correlation between the levels of Th1/Th2 cytokines in the serum before IFN treatment and the result of IFN therapy. Increasing IFN-gamma in the serum induced by IFN treatment is associated with sustained virological response.</p>

Selection and application of serotypical synthetic peptides derived from hepatitis C virus NS5A region / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Numerous studies have reported a relationship between hepatitis C virus (HCV) genotype and the response to interferon therapy. Despite high sensitivity and specificity, genotyping methods can be performed only on HCV RNA positive samples. Serotyping might be a rapid and cost effective method for determining HCV genotypes, especially in patients with previously undetectable HCV RNA. In this study, an enzyme linked immunosorbent assay (ELISA) method for HCV serotyping with the genotype specific, synthetic peptides derived from HCV nonstructural 5a (NS5A) region was developed.</p><p><b>METHODS</b>Based on 45 sequences, representing HCV genotypes 1 - 6 from Genebank, we synthesised 305 overlapping 30-mer peptides within NS5A region at positions 2182 - 2343 of HCV. All peptides for antigenic reactivity were tested by enzyme immunoassay with 69 human sera with antiHCV positive representing genotype 1 - 6. Forty hepatitis C patient sera were serotyped using serotype specific, synthetic peptides and genotyped by sequencing analysis.</p><p><b>RESULTS</b>The correspondence of amino acids in HCV NS5A region with amino acids in positions 2182 - 2343 was very low among different genotype peptides. The highly conserved sequences were residues 2182 - 2211 (R1), 2272 - 2301 (R7) and 2302 - 2331 (R9): the highly variable 2212 - 2241 (R3) and 2257 - 2286 (R6). Using 305 peptides, antigenic regions were located in R3, R7 and R9. Eighteen peptides from highly conserved region representing genotypes 1 to 6 showed broad immunoreactivity with sera containing antibody to all HCV genotypes. Immunoreactivity of the peptides from highly variable region was stronger with similar genotype sera. Twelve unique peptides showed highly, genotype specific, reactivity with types 1 and 3 sera. Type 2 genotype specific peptides had cross reaction with type 3 serum. No type 4, 5 or 6 specific peptides were selected. The serotyping results showed high agreement with sequencing analysis.</p><p><b>CONCLUSIONS</b>The major antigenic regions in HCV NS5A region were at 2212 - 2241 (R3), 2272 - 2301 (R7) and 2302 - 2331 (R9). Eighteen peptides from highly conserved region show genotype independent, immunoreactivity, useful for antiHCV antibody test. Twelve peptides from highly variable region show genotype 1 and 3 dependent immunoreactivity, useful for determining HCV serotype, especially for patients with previously undetectable HCV RNA.</p>

Mechanism of hepatitis B virus infection of trophoblast cells and hepatitis B virus intrauterine infection / 中华传染病杂志

1.0?10~7 copies/mL. The HBsAg IHC staining positive cells could be observed in 6 placental tissues and 3 fetus' liver tissues,and HBcAg was also positive in 1 case of fetus' liver tissue.After co-incubating the tropho- blastic cells and HBV DNA positive serum in vitro,HBsAg expression and HBV DNA could be detected.Apoptosis of HBV-infected trophoblastic cells increased,which was demonstrated by in vivo and in vitro experiments and the apoptosis of placental cells was correlated with the cord blood HBV DNA level.The results of in vitro experiments showed that the apoptosis of trophoblastic cells increased with the elongation of infection time.After 6 months,1 of 12 newborns was positive for HBsAg,HBeAg and anti-HBc,6 was positive for anti-HBs.Conclusions The mechanism of HBV intra-uterine infection may be that HBV breaches the placental barrier and infects the fetus.The localization and replication of HBV in fetal tissues and organs are probably the important factors of chronic HBV infections in neonates.The apoptosis of trophoblastic cells may be the protective mecha- nism for the placental barrier to block the HBV intra-uterine transmission.

Immunoreactivity studies on synthetic peptides deriving from immunodominant region of hepatitis C virus NS5a gene / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To identify the location of major immunodominant antigenic region and study the relationship between the gene heterogeneity and immunoreactivity via detecting antigenic reactivity of synthetic peptides deriving from immunodominant region in different genotypes of hepatitis C virus (HCV) NS5a gene.</p><p><b>METHODS</b>In total, 305 non-identical 30-mer long and overlapping by 15 aa peptides derived from HCV NS5a region from codon 2,182 to 2,343 among 45 unique published HCV sequences in GenBank corresponding to different genotype were designed and synthesized. The amino acid sequences of all peptides were compared with DNA Star software. The antigenic reactivity of those peptides was detected with indirect ELISA with both anti-HCV and anti-NS5 positive serum.</p><p><b>RESULTS</b>The sequences showed highly conserved among HCV genotype in regions 2,272-2,301 and 2,302-2,331 as compared to regions 2,212-2,241 and 2,257-2,286. The peptides basing on amino acid residues among 2,212-2,241, 2,272-2,301 and 2,302-2,331 showed stronger immunoreactivity than any other peptides. Eighteen peptides derived from this region showed a broad immunoreactivity, 3 of them could react with 96% of anti-HCV positive sera. Whereas the immunoreactivity of the peptides derived from the region showing highly variable among HCV genotype was found to react more strongly with homologous-genotype sera.</p><p><b>CONCLUSION</b>The major linear antigenic region was located at amino acid residues among 2,212-2,241, 2,272-2,301 and 2,302-2,331; the short synthetic peptide derived from NS5a region at position 2,212-2,241, 2,272-2,301 and 2,302-2,331 can be used for efficient detection of HCV antibody; some peptides showed genotype specific immuunoreactivity.</p>