Enhancement of T Follicular Helper Cell-Mediated Humoral Immunity Reponses During Development of Experimental Autoimmune Myasthenia Gravis / 神经科学通报·英文版

Myasthenia gravis (MG) is a prototypical antibody-mediated neurological autoimmune disease with the involvement of humoral immune responses in its pathogenesis. T follicular helper (Tfh) cells have been implicated in many autoimmune diseases. However, whether and how Tfh cells are involved in MG remain unclear. Here, we established and studied a widely-used and approved animal model of human MG, the rat model with acetylcholine receptor alpha (AChRα) subunit (R-AChR)-induced experimental autoimmune myasthenia gravis (EAMG). This model presented mild body-weight loss 10 days after the first immunization (representing the early stage of disease) and more obvious clinical manifestations and body-weight loss 7 days after the second immunization (representing the late stage of disease). AChR-specific pre-Tfh cells and mature Tfh cells were detected in these two stages, respectively. In co-cultures of Tfh cells and B cells, the number of IgG2b-secreting B cells and the level of anti-AChR antibodies in the supernatant were higher in the cultures containing EAMG-derived Tfh cells. In immunohistochemistry and immunofluorescence assays, a substantial number of CD4/Bcl-6 T cells and a greater number of larger germinal centers were observed in lymph node tissues resected from EAMG rats. Based on these results, we hypothesize that an AChR-specific Tfh cell-mediated humoral immune response contributes to the development of EAMG.

Research progress of tertiary lymphoid tissues in multiple sclerosis / 中国免疫学杂志

In recent years,it has been found that there are lymphoid follicle structures outside the lymphoid tissues and organs, which are called ectopic lymphoid tissues,also known as tertiary lymphoid tissues(TLTs).TLTs currently have been found in a number of diseases such as multiple sclerosis,diabetes,cancer,atherosclerosis and rheumatoid arthritis and the like.Here we focus on the molecular mechanism of TLTs and the relationship between TLTs and multiple sclerosis (MS)/experimental autoimmune encephalomyelitis (EAE) disease,and briefly describe the role of TLTs in the development of MS/EAE,the relevant mechanisms and research progress,make people have more understanding and insights on the formation of TLTs and the mechanism that it involved in disease,to provid theoretical knowledge for clinical treatment.

Role of interelenkin-17, -6 and endothelin-1 in statins attenuated no-reflow phenomenon of rat acute myocardial infarction and reperfusion / 中国地方病学杂志

ObjectiveTo study the relevant effect of proinflammatory cytokines interelenkin-17(IL-17), -6 and endothelin-1 (ET-1) on statins attenuating no-reflow phenomenon after myocardial ischemia-reperfusion in rats.MethodsEighteen healthy male Wistar rats were randomly divided into 3 groups according to body weight: sham operation, injury, preconditioning groups. The preconditioning group was given atorvastatin 2 mg·kg-1 ·d-1 and the other two groups were given the same volume of saline once. After 7 days, the rats were anesthetized with an intraperitoneal injection of chloral hydrate, and then the thoracic cavity was opened. The coronary artery of injury group and preconditioning group were ligated for 60 minutes, and then opened for 15 minutes, to establish the rat acute myocardial ischemia-reperfusion model. The sham operation group was was treated with a seam through the coronary artery without ligation. Eleetrocardiogram was checked before ligation, and ligation was carried out for 15, 30, 45 minutes and then reperfusion for 15 minutes. After reperfusion for 15 minutes, the thioflavine S and Even's were injected from femoral venous, then the heart and blood were obtained(keeping left ventricular only). Hearts were flushed with saline and sliced transversely into five to seven sections. Finally, observed at 365 nm wave length the existence of non-fluorescent areas, which was no-reflow zone. The level of serum IL-17, IL-6 and ET-1 was detected by ELISA. Results The electrocardiogram confirmed that the sham operation group had no ischemic damage and the model of myocardial ischemia- reperfusion was established in preconditioning group and injury group. The noreflow phenomenon could be observed under 365 nm wave length in preconditioning group and injury group. The ligated area[LA％, (57.34 ± 11.49)％, (53.08 ± 8.66)％] of injury group and preconditioning group was higher than that of sham operation group(0, all P ＜ 0.05); the area of no-reflow[ANF％, (48.96 ± 6.94)％, (21.37 ±3.35)％] of injury group and preconditioning group was higher than that of sham operation group(0, all P ＜ 0.05),and the ANF％ of preconditioning group was lower than that of injury group(P ＜ 0.05) ; the level of serum IL-17,IL-6 and ET-1[(151.67 ± 11.19) × 10-9, (167.89 ± 5.13) × 10-9, (322.37 ± 19.08) × 10-9 g/L] of injury group was higher than those of sham group and preconditioning operation group[(49.75 ± 14.06) × 10-9, (59.32 ± 5.26) ×10-9, (109.9 ± 12.12) × 10-9, (90.45 ± 11.63) × 10-9, (112.47 ± 10.40) × 10-9 and(198.91 ± 27.88) × 10-9 g/L,P ＜ 0.05], the level of serum IL-17, IL-6 and ET-1 of preconditioning group was higher than those of sham operation group(P＜ 0.05). Conclusionsno-reflow phenomenon is related with IL-17 and ET-1 which can promote the expression of IL-6, statins decreases the expression of IL-17 and ET-1, and then decreases the on-reflow phenomenon.

Effects of bone marrow mesenchymal stem cells on the proliferation of hepatocytes and cirrhotic fat-storing cells in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the differentiation of bone marrow mesenchymal stem cells (BMSCs) and the effects of BMSCs on the proliferation of cirrhotic fat-storing cells (CFSC) and hepatocytes in vitro.</p><p><b>METHODS</b>BMSCs and hepatocytes were isolated and harvested from the bone marrow and livers of rats. A co-culture system was set up by transwell inserts in which the two chambers were separated by a semipermeable membrane. BMSCs labeled with PKH26 were cultured with hepatocytes/CFSC in the co-culture system and also in a cell-cell direct contact culture system. Anti-albumin and anti-smooth muscle alpha-actin (alpha-SMA) antibodies were tested by using fluorescence immunocytochemistry. BMSCs and hepatocytes/CFSC cultured alone served as controls. The proliferation level of hepatocytes in the co-culture system was measured. CFSC were cultured with the conditional medium of BMSCs, and their quantities were measured microscopically.</p><p><b>RESULTS</b>Expression of albumin was observed in the hepatocytes of the two culture systems after they were cultured for 72 h but the albumin levels were higher in the cell-cell direct contact culture system (P<0.01). As compared to the controls, the number of hepatocytes was larger in the co-culture system (P<0.01). No expression of alpha-SMA in CFSC was observed in either culture system. The proliferation of CFSC was inhibited by the conditional medium of BMSCs. The longer the time of the co-culturing the more significant was the CFSC growth suppression (P<0.01).</p><p><b>CONCLUSIONS</b>BMSCs can be induced into hepatocytes by a local micro-environment formed by hepatocytes. BMSCs may promote proliferation of hepatocytes and inhibit proliferation of CFSC.</p>

Relationship between M3 receptor and myocyte apoptosis induced by acute myocardial infarction / 药学学报

<p><b>AIM</b>To explore the effects of M3 receptor on myocyte apoptosis induced by acute myocardial infarction in rats.</p><p><b>METHODS</b>Rat model was induced by ligation of the anterior branch of the left coronary artery. All animals were divided into four groups: sham-operated group, occlusion group, choline group (10 mg x kg(-1), iv), and 4DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) group (0.12 mg x kg(-1), iv). The serum malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined. The infarct size areas on the myocardium were identified by TTC staining. The apoptosis in cardiomyocyte was detected by TUNEL assay and apoptosis-related proteins in Bcl-2 and Fas expression were measured by immunohistochemistry assay.</p><p><b>RESULTS</b>M3 receptor agonist choline reduced serum MDA content and increased SOD activity. The myocardial expression of Bcl-2 was increased, whereas the expression of Fas was decreased by choline. However, blockade of M3 receptor by 4DAMP completely inhibited these effects of choline on cardiac myocytes.</p><p><b>CONCLUSION</b>Activation of M3 receptor has protective effect on myocyte apoptosis induced by acute myocardial infarction in rat, and this effect might be related to modulating the expression of some immediateearly genes including Bcl-2 and Fas.</p>

Protective effect of M3 receptor on H2O2 -induced apoptosis of rat myocardial cells in vitro / 药学学报

<p><b>AIM</b>To observe the effect of activation of M3 receptor on H2O2 induced apoptosis in cultured rat myocytes and to investigate its possible mechanisms.</p><p><b>METHODS</b>Isolated neonatal cardiomyocytes were cultured. Morphologic changes were observed by microscopy. The apoptosis in cardiomyocyte was detected by terminal deoxynucleotide transferase directed d-UTP nick and end labeling (TUNEL) assay. The expression of apoptosis-related protein in Bcl-2 and Fas was measured by immunohistochemistry assay. [Ca2+]i in single cardiomyocyte loaded with Fluo 3-AM was measured by confocal microscope.</p><p><b>RESULTS</b>H2O2-mediated myocyte apoptosis was attenuated by M3 receptor agonist choline (10 mmol x L(-1)). Pretreatment of cardiac myocytes with choline also increased Bcl-2, decreased Fas expression, and inhibited the increase in FI value of [Ca2+]i in H2O2-stimulated cardiac myocytes. However, blockade of M3 receptor by 4DAMP (10 nmol x L(-1)) completely inhibited the effects of choline on H2O2-stimulated cardiac myocytes.</p><p><b>CONCLUSION</b>Activation of M3 receptor showed protective effect on H2O2-induced apoptosis in cultured rat myocytes and this effect might be related to modulating the expression of some genes including Bcl-2 and Fas as well as the downregulation of [Ca2+]i.</p>