Oxidative stress in chronic hepatitis C patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study is aimed to investigate oxidative stress status in chronic hepatitis C (CHC) patients.</p><p><b>METHODS</b>52 CHC patients were divided into two groups according to the serum level of alanine aminotransferase (ALT): group A (elevated ALT group) and group B (normal ALT group). 20 healthy controls were included in this study. Serum levels of xanthine oxidase (XOD), malondialdehyde (MDA), oxidizided glutathione (GSSG), glutathione (GSH), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), glutathione reductase (GR) and vitamin C (Vc) were determined by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Serum levels of XOD, MDA, GST and GR increased in CHC patients compared with healthy controls. While, serum levels of GSH, GSH-Px and Vc decreased compared with healthy controls. Furthermore, serum levels of XOD, MDA, GSSG, GST and GR in group A were up-regulated compared with group B. Serum levels of GSH, GSH-Px and Vc in group A were down-regulated compared with group B. In CHC patients, serum ALT level positively correlated with serum levels of XOD, MDA, GSSG and GST, while, negatively correlated with serum levels of GSH, GSH-Px and Vc. Serum aspartate aminotransferase (AST) level positively correlated with serum levels of XOD, MDA, GSSG, GR and GST, while, negatively correlated with serum GSH-Px level in CHC patients. Serum gamma-glutamyl transpeptidase (GGT) level positively correlated with serum GR level and negatively correlated with serum GSH level in CHC patients. Serum alkaline phosphatase (AKP) level positively correlated with serum levels of MDA and GR in CHC patients. In CHC patients, serum XOD level was positively related with serum HCV RNA level.</p><p><b>CONCLUSION</b>Oxidative stress was increased in CHC patients. In CHC patients with elevated serum ALT level, oxidative stress usually became serious.</p>

Cellular toxicity and anti-tumor efficacy of iRGD modified doxorubixin loaded sterically stabilized liposomes / 药学学报

iRGD-modified sterically stabilized liposomes loaded doxorubicin (iRGD-SSL-DOX) were prepared and their cellular toxicity and anti-tumor efficacy were evaluated, comparing to doxorubixin loaded sterically stabilized liposomes (SSL-DOX) and RGD modified doxorubixin loaded sterically stabilized liposomes (RGD-SSL-DOX). The iRGD peptide, with both tumor targeting and cell penetrating functions, was conjugated to DSPE-PEG-NHS and DSPE-PEG-iRGD was obtained. DSPE-PEG-RGD was gained in the same way. iRGD-SSL-DOX, RGD-SSL-DOX and SSL-DOX were prepared by ammonium sulfate gradient method. The size and zeta potential of the liposomes were characterized by dynamic laser light scattering. The cellular toxicity study was done on B16 melanoma cell line and the anti-tumor efficacy study was carried on B16 cell line bearing C57BL/6 mice. The results showed that the particle sizes of liposomes were all around 90-100 nm. DOX entrapment efficiency was above 95%. The formulations were with good preparation reproducibility. iRGD-SSL-DOX showed no significant difference in B16 cellular toxicity with SSL-DOX and RGD-SSL-DOX, but the anti-tumor efficacy on B16 melanoma bearing C57BL/6 mice was significantly better than that of SSL-DOX, similar as that of RGD-SSL-DOX. Therefore, iRGD modified liposomes loaded DOX would be a promising drug delivery system for tumor therapy.

The analysis of IL-10 and its methylation in the patients with acute on chronic liver failure / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the effect of IL-10 and the methylation of its promoter in acute on chronic liver failure (ACLF).</p><p><b>METHODS</b>Patients were divided into three groups: 25 with ACLF, 25 with CHB, 10 healthy controls. Respectively detect the serum level of IL-10 via ELISA, and the methylation of IL-10 promoter via MSP, to analyze the difference among the three groups.</p><p><b>RESULTS</b>Both the ACLF group and the CHB group have significant increase in serum level of IL-10 compared with the control group (P < 0.05); the ACLF group's level is higher than the CHB group, however without statistical significance (P > 0.05). The serum level of IL-10 in ACLF group has no significant relativity with ALT and HBV-DNA( r = -0.022, r = 0.033, respectively; P > 0.05); has positive relativity with TBIL and MELD ( r = 0.566, r = 0.443, respectively; P < 0.05); and negative relativity with PTA (r = -0.581, P < 0.05). The distribution of the methylation of IL-10 promoter in ACLF group is significantly different from the other two.</p><p><b>CONCLUSION</b>The serum level of IL-10 in hepatitis patients is significantly higher and increases with the degree of liver failure. The promoter methylation may be important in the gene inactivation.</p>

Hypomethylation of TNF-alpha gene promoter in the patients with acute-on-chronic hepatitis B liver failure / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>The present study was designed to investigate the possible epigenetic alteration in the promoter of TNF-alpha in the patients with acute-on-chronic hepatitis B liver failure (ACHBLF).</p><p><b>METHODS</b>The methylation of TNF-alpha promoter in peripheral blood mononuclear cells (PBMCs) was measured by methylation specific PCR (MSP). The level of serum TNF-alpha was determined by enzyme-linked immunosorbent assay (ELISA). Model for End-stage Liver Disease (MELD) was performed for the evaluation of liver failure.</p><p><b>RESULTS</b>The serum level of TNF-alpha in patients with ACHBLF(44.9260 +/- 26.48523) was higher than that in CHB (18.92505 +/- 9.04461) and healthy controls (11.9172 +/- 5.04612) (P < 0.05). Moreover, the serum TNF-alpha level was significantly decreased in methylation group as compared to unmethylaiton group in patients with ACHBLF (P < 0.05). MELD was not significantly different between methylated and unmethylated group of ACHBLF patients (P > 0.05). In addition, the serum level of TNF-alpha was found to be positively correlated with serum total bilirubin (r = 0.891, P < 0.01) and MELD score (r = 0.792, P < 0.01), but to be negatively correlated with prothrombin activity (r = - 0.511, P < 0.05) in patients with ACHBLF.</p><p><b>CONCLUSION</b>The TNF-alpha methylation patten is stable for the liver failure, suggesting the effect of environment on methylation.</p>

Decreased expression of serum cortisol in patients with severe hepatitis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the profile of serum cortisol levels in patients with severe hepatitis.</p><p><b>METHODS</b>Fifty patients with viral hepatitis (including 30 severe hepatitis patients and 20 chronic hepatitis B patients) were enrolled in this present study. Serum cortisol concentration was tested using radioimmunoassay. Furthermore, liver function, coagulation and other related laboratory indices were also determined.</p><p><b>RESULTS</b>Serum cortisol concentration of severe hepatitis group was lower than that of chronic hepatitis B group (P < 0.05) and lower than that of healthy controls (P < 0.05) serum cortisol concentration of severe hepatitis patients was significantly positively correlated with PTA (r = 0.445, P < 0.05); serum cortisol concentration has no relation with ALT in patients with severe hepatitis (P > 0.05), and serum cortisol concentration was significantly negatively correlated with the ratio of AST/ALT in patients with severe hepatitis (r = -0.367, P < 0.05). No significant relationship was found between serum cortisol concentration and total Bilirubin (P > 0.05). Serum cortisol concentration in death group of severe hepatitis was lower than that in survival group of severe hepatitis (P < 0.05). Of severe hepatitis patients with MELD score, the higher MELD score, the lower the cortisol concentration.</p><p><b>CONCLUSION</b>Cortisol concentration decreased in patients with severe hepatitis, which was related to functional liver reserve and disease severity. Cortisol can be related to the prognosis of severe hepatitis patients.</p>

Study of oxidative stress in chronic hepatitis B patients with elevated serum total bilirubin / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate oxidative stress in chronic hepatitis B (CHB) patients with elevated serum total bilirubin (TBIL).</p><p><b>METHODS</b>75 CHB patients with elevated serum TBIL were enrolled in the present study. A, B, C, D and E group were defined. Serum Malondialdehyde (MDA), Xanthine Oxidase (XOD), Vitamin C (V(C)) and Vitamin E (V(E)) were determined. The control group contained 11 healthy donors and the carrier group contained 16 Hepatitis B surface antigen (HBsAg) carriers.</p><p><b>RESULTS</b>The concentrations of MDA and XOD were significantly higher in each group of patients than in the control (P < 0.05), while V(C) and V(E) were significantly lower (P < 0.05). The concentration of XOD was significantly higher in the carrier group than in the control (P < 0.05), while MDA, V(C) and V(E) were not significantly different (P > 0.05). The concentrations of MDA and XOD were significantly positively correlated with TBIL (r = 0.670, P < 0.01; r = 0.737, P < 0.01, respectively) in the patients, while V(C) and V(E) were significantly negatively correlated with TBIL (r = -0.463, P < 0.01; r = -0.247, P < 0.05, respectively). The concentration of MDA was significantly different among all the groups in the patients except the comparison between group A and group B. The concentration of XOD was significantly different between group A, B, C and group D, E (P < 0.05). The concentration of V(C) was significantly different between group A and group D, E and between group B, C, D and group E (P < 0.05). The concentration of V(E) was significantly different between group A, B and group E (P < 0.05).</p><p><b>CONCLUSION</b>There was a disturbance between oxidative stress and anti-oxidative ability in CHB patients with elevated serum TBIL. Oxidative stress became more serious along with the increasing of serum TBIL. In HBsAg carriers, oxidative stress level was low. The results suggest antioxidant treatment for CHB patients with elevated serum TBIL may help to improve the effect of therapy.</p>

Study on the relationship of serum vitamin E and liver pathological features in the patients with chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study on the relationship of serum vitamin E and liver pathological features in the patients with chronic hepatitis B.</p><p><b>METHODS</b>Sixty-six patients with chronic hepatitis B and ten healthy controls were enrolled in this present study. The serum vitamin E level was measured spectrophotometrically. Comparisons of liver function test, HBeAg and HBV DNA level were conducted among different liver pathological features including inflammatory grading and fibrosis staging.</p><p><b>RESULTS</b>Compared with healthy controls, the serum level of vitamin E was significantly decreased in the patients with chronic hepatitis B, especially in those with elevated ALT activity. In comparison between HBeAg positive group and HBeAg negative group, the serum level of vitamin E of the former group did not significantly changed (P > 0.05). Furthermore, the serum level of vitamin E has been demonstrated to be negatively associated with the inflammation grading in the patients with chronic hepatitis B. However, there was no significant association between the serum vitamin E and liver fibrosis staging.</p><p><b>CONCLUSION</b>Vitamin E, as one of the important anti-oxidants, was demonstrated to be implicated in the progression of liver inflammation in the patients with chronic hepatitis B. Furthermore, the supplement of vitamin E would be a potential therapy for attenuate the inflammatory response.</p>

Increased expression of inducible nitric oxide synthase in chronic hepatitis B patients is correlated with histopathological grading and staging / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the intrahepatic expression of inducible nitric oxide synthase (iNOS) in patients with chronic hepatitis B (CHB) and its relation to liver histopathology.</p><p><b>METHODS</b>The intensity and distribution of the immunohistochemical staining of intrahepatic iNOS were studied in the liver biopsy specimens obtained from 74 patients with CHB and statistical analyses were performed between intrahepatic iNOS and ALT, HbeAg, HBV DNA grading of liver inflammation and staging of fibrosis. Seven histologically normal liver sections were used as a control group.</p><p><b>RESULTS</b>Compared with the control group, the intrahepatic iNOS immunoexpression was significantly higher in patients with CHB (P < 0.05), iNOS immunoreactivity was observed mainly in hepatocytes showing a predominant cytoplasmic staining, with the positive liver cells distributed diffusely throughout the hepatic lobule. Immunopositive staining could also be detected in Kupffer cells, sinusoidal lining cells and vascular endothelial cells. Compared with patients with normal ALT, the hepatocellular iNOS immunoexpression was significantly higher in patients with elevated ALT (P < 0.05) and the iNOS immunoexpression was significantly correlated with the serum level of ALT (r=0.601, P=0.000). Statistical analysis also showed that the intrahepatic iNOS immunoexpression was positively correlated with the grading of liver inflammation and the staging of liver fibrosis (r=0.660, P=0.000; r=0.507, P=0.000). No significant correlation between iNOS and HBeAg and HBV DNA was detected. CONCLUSION The intrahepatic expression of iNOS is elevated in chronic hepatitis B patients and correlated well with the severity of the disease, which indicated that inducible nitric oxide synthase may have a critical role in the pathogenesis of chronic viral hepatitis B.</p>

Study of serum level of cortisol and peripheral T lymphocyte subsets state in the hepatitis B virus carriers / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study of serum level of cortisol and peripheral T lymphocyte subsets state in the hepatitis B virus (HBV) carriers.</p><p><b>METHODS</b>Sixty chronic HBV carriers and ten healthy controls were all enrolled in this present study. Serum expression of cortisol was determined by radioimmunoassay, and also flow cytometry was performed to evaluate peripheral blood T lymphocyte subset.</p><p><b>RESULTS</b>Compared with those in normal controls, the serous levels of cortisol in chronic HBV carriers were significantly elevated, while there was no distinct difference in the proportion of CD4+ T lymphocytes ( P > 0.05) with the decreased odds of CD4+/CD8+ lymphocytes( P < 0.05) and obvious higher proportion of CD8+ T lymphocytes( P < 0.05). In comparison between HBeAg positive group and HBeAg negative group, the serous levels of cortisol of the former group were significantly higher ( P < 0.05), and so proportion of CD8+ T was too ( P < 0.05). However, there is no significant differences in the proportion of CD4+ T lymphocyte ( P > 0.05).</p><p><b>CONCLUSION</b>The elevated serum cortisol and increased CD8+ T lymphocytes subsets in the chronic HBV carriers, suggested that there was disturbance of endocrine-immune response in the chronicity of HBV infection.</p>

Oxidative stress in patients with chronic hepatitis B before and after interferon alpha-2b treatment / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the impacts of interferon alpha-2b (IFN alpha-2b) on the oxidative stress states in the treatment of chronic hepatitis B (CHB) with different genotypes.</p><p><b>METHODS</b>Thirty-five patients with chronic hepatitis B and 18 healthy volunteers as a control were enrolled in this present study. In control and patients group, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), serum malondialdehyde (MDA) levels, serum total antioxidative stress capacity (TAC) were measured spectrophotometrically. After the therapy with interferon alpha-2b at the dose of 300 million units via intramuscular injection thrice a week for 12 weeks, these parameters were measured again in the patient group. The genotypes of hepatitis B virus were detected by polymerase chain reaction and hybridization. The effective group was defined as the patients with complete response and partial response.</p><p><b>RESULTS</b>The elevated concentrations of MDA and impaired levels of TAC in the patients with CHB were observed as compared to the healthy controls (P < 0.05 for both). There were no significant differences in serum levels of MDA and TAC in CHB patients with various genotypes (P > 0.05). The serum levels of MDA after the treatment with IFN alpha-2b were significantly lower than the pretreatment levels (P < 0.05), which even returned to the normal concentration (P > 0.05) in the effective group. There were significant increases in the TAC after the IFN alpha-2b therapy in the effective group. However, the significant differences in the TAC levels before and after the INFalpha-2b treatment were not observed in the non-responsive group.</p><p><b>CONCLUSION</b>The oxidative stress could be improved with IFN alpha-2b treatment of chronic hepatitis B patients. The results suggest that antioxidant treatment for chronic hepatitis B patients may help improve the effect of anti-virus therapy.</p>