Effect of Stains on the Purity Determination of Human Serum Albumin by Agarose Gel Electrophoresis / 中国药学杂志

OBJECTIVE: To study the effect of stains on the purity detection results of human serum albumin by agarose gel electrophoresis, thus to select appropriate stain so that the detection result of agarose gel electrophoresis method can be basically consistent with the method specified in the 2015 edition of Chinese Pharmacopoeia, cellulose acetate membrane electrophoresis method. METHODS: Acid blue, amino black and ponceau were used in agarose gel electrophoresis to detect the purity of 30 batches of human serum albumin samples, and the results were compared with those obtained by cellulose acetate membrane electrophoresis. RESULTS: Impurity bands could not be detected by ponceau stain; the results of amino black staining was significantly lower than that of cellulose acetate membrane electrophoresis. There was no statistical difference between the results of amino black staining and cellulose acetate membrane electrophoresis. The results of 25 batches of samples detected by cellulose acetate membrane electrophoresis were within the 95% confidence interval of the results detected by amino black staining. CONCLUSION: The category of stain has great influence on the detected purity of human serum albumin when using agarose gel electrophoresis. The results of amino black staining is consistent with those of the method recorded in the Chinese Pharmacopoeia in use.

Effects of blueberry on hepatic fibrosis and expression of nuclear transcription factor-кB in rats / 中华肝脏病杂志

Objective@#To observe the effects of blueberry and nuclear expression of transcription factor-кb (NF-кb) p65 in an experimental rat model of liver fibrosis.@*Methods@#Forty-five Sprague-Dawley rats were randomly divided into isotonic saline control group (A); model group (B); blueberry juice prevention group (C, 15 g/kg); dan-shao-hua-xian capsule prevention group (D, 1 g/kg); and blueberry juice + dan-shao-hua-xian capsule prevention group (E). Rat liver fibrosis model was established by covalent compound carbon tetrachloride (CCl4). Each prevention group was given the corresponding dose of blueberry juice or (and) dan-shao-hua-xian capsule, and the rats were sacrificed after 8 weeks. The degree of liver fibrosis was evaluated by hematoxylin and eosin stain. A liver tissue of NF-κBp65 was detected by immunohistochemical method. The NF-κBp65 protein expression of liver tissue and transforming growth factor (TGF) β1 was detected by Western blot. Data of multiple groups were compared by one-way analysis of variance, and rank sum test.@*Results@#Immunohistochemistry detected that TGFβ1 protein was mainly expressed in mesenchymal origin of hepatic stellate cells. The expression level of group A (3.75 ± 1.67) was low, while those of group B (9.00 ± 2.07), C (7.33 ± 1.00), D (6.00 ± 1.51), and E (3.5 ± 1.41) were high. However, the expression level of TGF-β1 protein in hepatic tissues of group B was significantly higher than that of group C, D and E [group E: 3.5 ± 1.41, F = 18.350, P < 0.05]. In addition, group D was higher than group E (P < 0.05). The expression of NF- kappa Bp65 protein was very complex, and the expression patterns in different groups were different (Statistical calculation of experimental data were based on expression of liver cells). Compared with group B (4.37 ± 2.13), the relative expression levels of NF-κBp65 protein in-group A (0.46 ± 0.25), group C (2.76 ± 1.01), group D (2.13 ± 1.51), group E (1.88 ± 0.99) were significantly decreased (F = 27.490, P < 0.05), and the expression trend was consistent with TGFβ1 protein. Western blot detected NF-κBp65 protein in liver tissues of rats. Compared with group A, levels in groups B, C, D and E were significantly increased (F = 96.983, P < 0.05), and groups C, D and E were significantly lower. The E group was significantly lower than the C group (F = 96.983, P < 0.05), and the degree of hepatic fibrosis was lower in each prevention group than in the B group (T = 24.1, P < 0.05).@*Conclusion@#Blueberries have preventive effect on CCl4-induced hepatic fibrosis in rats, and its preventive mechanism may inhibit the expression and activation of NF-κBp65 in hepatocytes, thereby reducing TGFβ1- mediated production or activation.

Natural clearance of hepatitis C virus in 96 patients with infection acquired by blood transfusion from a single donor in Guizhou / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the clinical features and rate of natural viral clearance in patients with hepatitis C virus (HCV) infection acquired by blood transfusion from a single donor.</p><p><b>METHODS</b>Ninety-six patients who acquired HCV infection between January 1998 and December 2002, upon receipt of donated blood from a single infected individual in Guizhou,were included in this retrospective cross-sectional study. Patients were clinically assessed to determine levels of anti-HCV antibodies, HCV RNA and biochemical indicators of liver function,as well as features of liver structure (by abdominal B ultrasonography and elastography). HCV genetic testing was used to determine the virus genotype. Measurement data were expressed as mean ± standard deviation. Count data were analyzed by the x² test,with P less than 0.05 indicating statistical significance.</p><p><b>RESULTS</b>All 96 patients tested positive for antiHCV antibodies. The majority of patients (70%; 34:33 male:female) had HCV RNA more than or equal to 1.0 * 103 copies/ml. All patients carried the same HCV genotype as the single blood donor:genotype lb. The overall rate of natural HCV clearance was 30.2%. but males had a significantly lower rate (19.0% (8/42) vs. females:38.9% (21/54);x²=4.41,P=0.023) as did older patients (more than 40 years-old:16.1% (5/31) vs .less than or equal to 40 years-old:36.9% (24/65);x²=4.30,P=0.028). The overall rate of chronic HCV infection (CHC) was 69.8%,but the rate was significantly lower in younger patients (less than or equal to 40 years-old:63.1% (41/65) vs. more than 40 years-old:83.9% (26/31);x²=6.67,P=0.028). Among the 67 patients with CHC,12 had symptoms of mild weakness,anorexia and abdominal distention,11 had elevated serum alanine aminotransferase (116.25 +/- 24.65 U/L) and stage 3 or 4 fibrosis (liver elasticity values more than or equal to 5.1 kPa),and three had mildly abnormal serum bilirubin (32.56 ± 5.28 mumol/L). Fifteen patients showed signs of chronic hepatitis and one patient showed signs of cirrhosis by abdominal B ultrasonography. None of the patients showed signs of hepatocellular carcinoma.</p><p><b>CONCLUSION</b>The course of blood transfusion acquired HCV infection is largely unknown and natural viral clearance rate may be associated with sex-and age-related factors.</p>

Effects of diterpene phenol extract of Rosmarinus officinalis on TGFbeta1 and mRNA expressions of its signaling pathway molecules in the lung tissue of pulmonary fibrosis rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the regulative mechanism of the diterpene phenol extract of Rosmarinus Officinalis (DERO) on the imbalance of collagen metabolism of the lung tissue in pulmonary fibrosis rats.</p><p><b>METHODS</b>Fifty healthy Sprague-Dawley rats were randomly divided into the normal saline group (NS), the bleomycin-induced lung injury group (BLM), the low dose DERO group (at the daily dose of 50 mg/kg), the moderate dose DERO group (at the daily dose of 100 mg/kg), and the high dose DERO group (at the daily dose of 200 mg/kg), 10 in each group (abbreviated as DERO 1, 2, 3, respectively). The pulmonary fibrosis rat model was prepared by disposable intratracheal instillation of bleomycin. DERO was administered by gastrogavage as intervention during the repairing process of lung injury. On the morning of the 29th day, the rats' lung tissue was extracted. The karyocyte number, collagen protein, type I collagen (collagen I) and transforming growth factor-beta type II receptor (TGFbetaR II), Smad4 mRNA expressions were semi-quantitatively determined using tissue microarray, HE staining, collagen fiber dyeing, immunohistochemical assay, and in situ hybridization. Using real-time fluorescent quantification RT-PCR, the mRNA expression of transforming growth factor-beta1 (TGF-beta1) were detected.</p><p><b>RESULTS</b>Compared with the NS group, the collagen deposition of the lung tissue was obvious and the inflammatory infiltration was more severe in the BLM group (P < 0.05, P < 0.01). There was no statistical difference in the aforesaid 4 indices between the DERO1 group and the BLM group (P > 0.05). The collagen deposition and the inflammatory infiltration were obviously alleviated in the DERO2 and DERO3 groups (P < 0.05, P < 0.01). Compared with the NS group, the mRNA expressions of collagen-I, TGF-beta1 R II, Smad4, and TGF-beta1 were obviously up-regulated in the BLM group (P < 0.05, P < 0.01). Compared with the BLM group, the aforesaid four indices were not statistically changed in the DERO1 group (P > 0.05). But the mRNA expressions of collagen-I, TGF-beta1 R II, Smad4, and TGF-beta1 were obviously downregulated in the DERO2 and DERO3 groups (P < 0.05, P < 0.01). But the down-regulation of Smad4 expression was not obvious in the DERO2 and the DERO3 groups (P > 0.05). Compared with the DERO1 group, the mRNA expressions of collagen-I, TGF-beta1, R II, TGFbeta1 were all obviously lower in the DERO2 and the DERO3 groups (P < 0.05). But there was no statistical difference in the aforesaid 4 indices between the DERO2 group and the DERO3 group (P > 0.05).</p><p><b>CONCLUSIONS</b>DERO could regulate imbalanced collagen metabolism of pulmonary fibrosis. It could inhibit excessive deposition of collagen fibers, especially excessive deposition of collagen- I. Its mechanisms might be realized by inhibiting up-regulation of TGF-beta1 and TGFbetaR II mRNA expressions, thus interfering the activation of TGF-beta-Smad signaling pathway on target genes, especially on type I procollagen target gene.</p>

Investigation of antiviral efficacy in patients with chronic hepatitis C originating from a same blood donor in Guizhou Province / 中华传染病杂志

Objective To investigate the antiviral efficacy of standard treatment with interferon (IFN)-α 2b and ribavirin (RBV) in patients with chronic hepatitis C (CHC) originating from a same blood donor.Methods The test group consisted of 65 CHC patients originating from a same blood donor,and was treated with IFN-α 2b 3-5 MU every other day in combination with RBV 0.6-1.0 g/d.Meantime,the control group consisted of 32 CHC patients who visited the Department of Infectious Diseases in Qiannan People's Hospital,and was treated with Peg-interferon (PEG-IFN)-α 2a 180 μg every week in combination with RBV 0.6-1.0 g/d.All the patients in the two groups were treated for 48 weeks and followed up for 96 weeks.Assessment indictors included sustained virological response (SVR),early virological response (EVR),end of treatment virological response (ETVR),biochemical response after withdrawal of treatment.Side effects during treatment were also evaluated.Measurement data were analyzed by x2 test.Results In test group,SVR rate was 83.1％ (54/65),EVR rate was 93.8％ (61/65),ETVR rate was 86.2％ (56/65) and biochemical response rate after withdrawal of treatment was 100.0％.In control group,SVR rate was 87.5％ (28/32),EVR rate was 96.9 ％ (31/32),ETVR rate was 90.6 ％ (29/32) and biochemical response rate after withdrawal of treatment was 100.0 ％.SVR rates of the two groups were not significantly different (x2 =0.072,P=0.086).Patients of the two groups were divided into two subgroups according to viral load:hepatitis C virus (HCV) RNA＜1.0 × 106 copy/mL and HCV RNA≥1.0 × 106 copy/mL.SVR rates of patients with low and high viral load in test group were 88.9％ and 54.5％,respectively (x2=7.67,P=0.008),those in control group were 96.0％ and 57.1％,respectively (x2 =4.41,P=0.038).SVR rates were higher in the subgroup of patients with low viral load.Leukopenia and thrombocytopenia were more common in control group than in test group (x2 =9.805,P =0.003 ; x2 =6.643,P=0.009).Conclusion IFN-α 2b and RBV combination therapy has similar antiviral efficacy to that of PEG-IFN-α 2a and RBV combination therapy,and has a lower rate of side effects as well.