Synchronization isolation method for multiple types of cells from mouse liver / 中华肝脏病杂志

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.

Epidemiological study and intervention measures of acute respiratory virus infection in hospitalized patients / 公共卫生与预防医学

Objective To understand the epidemiological characteristics of acute respiratory virus infection in hospitalized patients, and to provide reference for clinical diagnosis and treatment, and to develop relevant intervention measures. Methods A total of 414 hospitalized patients with respiratory virus infection admitted to our hospital from March 2019 to March 2021 were selected. Immunofluorescence method was used to qualitative detect parainfluenza virus type 1-3, influenza A virus, respiratory syncytial virus (RSV), influenza B virus and adenovirus. Results Among the 414 ARTI patients, 84 cases were positive for respiratory virus, with a positive detection rate of 20.29% (84/414 ) . 76 cases were positive for single virus infection, with a positive detection rate of 18.36% (76/414) . The positive detection rate was 1.93% (8/414). The most common pathogens of virus infection were influenza A virus (25.00%), influenza B virus (20.23%) and RSV(17.86%). There was statistical significance in the positive rate of ARTI among different age groups (χ2, P2, P0.05). The positive rate of PIV3 was the highest in spring (4.04%), and the positive rate of RSV in spring and winter was 24.24% and 25.20%, respectively. The positive rates of influenza A virus and influenza B virus were the highest in winter (9.45%) and (7.09%). There was statistical significance in the positive rate of ARTI among different clinical diagnoses (χ2, P<0.05). The positive rates of PIV2, PIV3, influenza A virus and influenza B virus were significantly different (P<0.05). The positive rate of bronchopneumonia virus was the highest (27.48%). Conclusion: RSV infection is the most common in patients with acute respiratory virus infection in Chengdu area, which mostly occurs in autumn and winter, and the main clinical manifestation is bronchopneumonia. The main infected population is children under 8 years old, and the surveillance of respiratory syncytial virus should be strengthened in the future.

HBeAg seroconversion achieved by sequential peginterferon alfa-2a therapy in chronic hepatitis B patients with unsatisfactory end point following entecavir treatment / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and safety of peginterferon alfa-2a (Peg-IFNa-2a) therapy for treating chronic hepatitis B (CHB) in patients who failed to achieve a satisfactory end point with entecavir (ETV) treatment.</p><p><b>METHODS</b>Fifty-seven CHB patients with positivity for hepatitis B e antigen (HBeAg) who had completed a standard ETV monotherapy course, of at least 96 weeks, and who had achieved a virological response (defined as HBV DNA less than 500 copies/ml) but without HBeAg seroconversion (defined as 0.227 PEI U/ml less than HBeAg less than or equal to 50 PEI U/ml) were enrolled in the study. The patients were randomly assigned to receive a 48-week treatment with Peg -IFNa-2a (experimental group, n = 27) or continued ETV therapy (control group, n = 30). Serum samples were collected from all patients for assessment of biochemical, virological and serological responses to treatment. Inter-group differences were statistically evaluated by t-test or Chi-squared test.</p><p><b>RESULTS</b>The baseline levels of alanine aminotransferase, hepatitis B surface antigen (HBsAg), and HBeAg were similar between the patients comprising the experimental and controls groups. At treatment week 48, the experimental group showed significantly higher rates of HBeAg clearance (Peg-IFNa-2a: 40.7% vs. ETV: 16.7%, x2 = 4.079, P less than 0.05) and seroconversion (37.0% vs. 13.3%, x2 = 5.110, P less than 0.05). The experimental group also showed higher rates of HBsAg clearance (7.4% vs. 0%) and HBV DNA relapse (11.1% vs. 0%), but the differences did not reach statistical significance (x2 = 2.307 and 3.519, both P more than 0.05). However, the level of HBsAg was significantly lower in the experimental group (2866.0+2580.4 vs. 4335.8+2650.0 IU/ml, t = 5.11, P less than 0.05).</p><p><b>CONCLUSION</b>HBeAg-positive CHB patients with unsatisfactory response to initial ETV monotherapy achieved HBeAg seroconversion and clearance following sequential Peg-IFN a-2a treatment.</p>

Colocalization of 11beta-hydroxysteroid dehydrogenase type I and glucocorticoid receptor and its significance in rat hippocampus / 生理学报

This paper was designed to observe the colocalization of 11beta-HSD1 and GR, and its significance in the rat hippocampus. Immunocytochemical dual-staining showed that not only 11beta-HSD1 but also GR immunoreactive substances were present in the cultured rat hippocampal neurons. Moreover, they were colocalized in the same hippocampal neuron. Synthetic glucocorticoid dexamethasone (DEX) up-regulated the protein expression and activity of 11beta-HSD1 in the cultured hippocampal neurons, as determined by Western blot and thin layer chromatography (TLC) respectively. The transfection of PC12 cells with the plasmid containing promoter sequence of 11beta-HSD1 gene and the reporter gene of CAT enzyme was conducted. DEX up-regulated the reporter gene expression in the system described above. The up-regulation of 11beta-HSD1 and reporter gene expression induced by DEX were both blocked by GR antagonist RU38486. Our study suggests that the colocalization of 11beta-HSD1 and GR in the hippocampus may be implicated in the up-regulation of 11beta-HSD1 expression by glucocorticoids combining to its promoter region, which in turn produces more biologically active glucocorticoids necessary for the binding of low affinity of GR.