目的 确定CD4⁺CD25⁺Treg调节性T细胞在重症肺炎克雷伯菌肺炎中的表达以及意义,探讨CD4⁺CD25⁺Treg在重症肺炎克雷伯菌肺炎的免疫抑制中的调控作用。方法 通过气管内滴注肺炎克雷伯菌菌液建立重症肺炎模型。采用流式细胞仪检测CD4⁺CD25⁺Treg细胞及酶联免疫吸附法(ELISA)等方法检测各种细胞因子。结果 重症肺炎克雷伯菌肺炎大鼠的脾脏和肺中CD4⁺CD25⁺Treg的数量增加。使用了CD25抗体(PC61)去除机体内源性的CD4⁺CD25⁺Treg,分别去除脾脏和肺的94%和90%的CD4⁺CD25⁺Treg。CD25抗体组在建模4 h,12 h及24 h后,肺部MPO及血清IL-1,IL-6,MIP-2较对照组高(P<0.05),肺和BLA比对照组高(P<0.05),CD25抗体组大鼠生存率比对照组低(P<0.05)。结论 内源的CD4⁺CD25⁺Treg对大鼠抑制重症肺炎克雷伯菌肺炎的过度免疫损害反应起到保护作用。

Objective To confirm the expression and meaning of the T regular cell in the severe Klebsiella pneumonia, and to evaluate the regular and control affect in the immunologic suppression of the severe Klebsiella pneumonia. Methods To build the severe pneumonia model by intratracheally inoculated with Klebsiella pneumoniae bacteria. To check sorts of inflammation factors by the methods of ELISA and flow cytometry. Results The quantity of the CD4⁺CD25⁺Treg in the splenic and lungs of the mice with severe Klebsiella pneumonia were increased. Anti-CD25Ab(PC61) was used to remove endogenousCD4⁺CD25⁺Treg. Anti-CD25 treatment remove 90% of CD4⁺CD25⁺Treg cells. The cytokine production(IL-1β,IL-6,MIP-2)in the anti-CD25-treated group were significantly increased. And it also increased significantly in the airway neutrophil infiltration, while the survival rate had been decreased. Conclusion Endogenous CD4⁺CD25⁺Treg can provide obvious protection effect to the restraining the over immunity damage of the severe Klebsiella pneumonia for the mice.

目的 水解乳清蛋白对炎症性肠病大鼠的抗炎作用及机制。方法 将40只雄性大鼠随机分为实验组和对照组,并建立炎症性肠病动物模型,分别喂食添加了水解乳清蛋白及普通蛋白的饲料,喂养4周后处死大鼠,每周检测体重,血清ALB、TNF-α、IL-2、IL-6等。结果 二组间体重及血清白蛋白无区别(P>0.05),实验组与对照组的TNF-α、IL-2及IL-6无区别(P>0.05),从第二周到第四周,实验组的炎症因子水平低于对照组(P<0.05)。结论 水解乳清蛋白具有抗炎作用,能够减少炎症性肠病大鼠动物模型的炎症因子的释放,并改善其营养状况。

Objective To evaluate the anti-inflammatory effects of whey protein on SD rats model of inflammatory bowel disease. Methods 40 SD rats model of inflammatory bowel disease were established and randomly divided into experimental and control groups equally. Experimental and control groups were fed whey protein and ordinary protein respectively. After 4 weeks, TNF-α, IL-2 and IL-6 were detected. Results There were no significant difference between the two groups of weights and the level of ALB. The level of TNF-α, IL-2 and IL-6 between groups were not significantly different in the first week(P>0.05). However, thelevels of TNF-α, IL-2 and IL-6 in experimental group were significantly lower than those of the control group in the follow weeks. Conclusion The whey protein could reduce the production of inflammatory cytokines.

目的 观察Bax抑制因子(Bax-inhibiting peptides,BIP)对肠上皮细胞的保护作用,并探讨其作用机制。方法 建立1日龄新生大鼠的坏死性小肠结肠炎模型。BIP于建模前腹腔内注射,建模后分别检测各组肠组织病理,并分别用流式细胞仪检测各组肠细胞凋亡率,western blot法检测bax下游凋亡蛋白细胞色素C、caspase 9和caspase 3含量。结果 与NEC组及Bax10 μg组比较,bax50 μg和100 μg可减轻肠上皮损伤,减少肠细胞凋亡率,降低bax下游凋亡蛋白细胞色素C、caspase9和caspase3含量。结论 一定剂量的Bax抑制肽可通过降低活性Caspase-3及CytC蛋白释放,保护线粒体膜,抑制肠上皮细胞调亡,发挥对肠上皮细胞的保护作用。

Objective To investigate the protective effects of Bax-inhibiting peptides(BIP) on intestinal epithelial cells and to explore its mechanism. Methods The model of neonatal necrotizing enterocolitis in 1 day neonatal rats were established. Before establishing the model, BIP was intraperitoneal injected to the rats. The pathological of intestinal tissue were detected respectively, the intestinal cell apoptosis rate were detected by flow cytometry, the levels of downstream apoptosis proteins of Bax were detected by Western blotting respectively. Results Compared to NEC and Bax10μg group, bax50μg and 100 μg can significantly reduce the intestinal epithelial damage and intestinal cell apoptosis rate and can decrease the levels of cytochrome C, caspase-9 and caspase-3, downstream apoptosis proteins of Bax,significantly. Conclusion A certain dose of Bax-inhibiting peptides can protect the mitochondrial membrane, inhibit the apoptosis of intestinal epithelial cells by reducing the level of Caspase-3 and CytC and play a protective effect on intestinal epithelial cells.

目的 建立大鼠急性心肌梗死缺血再灌注后无复流模型,并初步验证细胞焦亡在其中的发生情况。方法 选用20只标准成年雄性Sprague Dawley大鼠（体质量260~320 g）,随机分为对照组（n=5）和手术组（n=15）。对照组仅穿线冠状动脉,未行结扎;手术组结扎左前降支0.5 h后解除,进行再灌注4 h,以建立无复流模型。通过Evens blue和硫磺素S染色,评估心肌的正常供血区、再灌注区及无复流区,并对两组大鼠心肌组织进行病理分析。结果 对照组大鼠全部存活,未出现无复流现象,心肌组织中未见细胞焦亡。手术组存活13只,形成明确的正常供血区（n=13）、再灌注区（n=13）和无复流区（n=10）。在无复流区的心肌细胞中均观察到细胞焦亡（n=10）,而正常供血区未见（n=0）,再灌注区部分出现（n=4）,差异具有统计学意义（P<0.05）。结论 细胞焦亡现象主要存在于大鼠急性心肌梗死缺血再灌注后无复流区域中,细胞焦亡可能作为一种区域特异性程序性死亡方式,在心肌无复流的发生与发展中发挥重要作用。

Objective To establish a rat model of myocardial no-reflow after acute myocardial infarction with ischemia-reperfusion injury and to preliminarily explore the occurrence of pyroptosis in the affected myocardium. Methods Twenty adult male Sprague-Dawley rats（260-320 g）were randomly divided into a control group（n=5）and a surgical group（n=15）. In the control group,the coronary artery was encircled with suture but not ligated. In the surgical group,the left anterior descending artery was ligated for 30 minutes, followed by 4 hours of reperfusion to induce the no-reflow model. Evans blue and thioflavin S staining were used to evaluate the normal perfusion area,reperfusion area,and no-reflow area of the myocardium. Histopathological analysis was conducted on myocardial tissues from both groups. Results All rats in the control group survived without evidence of no-reflow or pyroptosis in myocardial tissue. In the surgical group, 13 rats survived and showed distinct regions of normal perfusion, 13 with reperfusion, and 10 with no-reflow. Pyroptosis was observed in all no-reflow areas（n=10）, absent in the normal perfusion zones（n=0）, and partially present in the reperfusion zones（n=4）. The differences were statistically significant（P<0. 05）. Conclusions Pyroptosis predominantly occurs in the no-reflow zones following acute myocardial infarction and ischemia-reperfusion injury in rats. As a region-specific form of programmed cell death, pyroptosis may play an important role in the development of myocardial no-reflow.

目的 初步探究胆管结扎诱导的梗阻性胆汁淤积对大鼠肝细胞的影响。方法 10只Lewis大鼠随机分为对照组和胆汁淤积组,每组各5只,胆汁淤积组采用胆管结扎2周诱导梗阻性胆汁淤积大鼠模型。苏木精-伊红染色和苯胺蓝染色比较组织病理变化,使用生化分析比较两组小鼠肝功能情况。采用改良的两步胶原酶灌注分离原代肝细胞,通过RT-qPCR检测两组小鼠肝细胞标志基因、细胞增殖标志基因以及胆管细胞标志基因的表达情况。结果 与对照组相比,胆汁淤积组肝脏表现为明显的肝组织紊乱和纤维胶原蛋白沉积以及肝功能的损害。胆汁淤积组较对照组的原代肝细胞更高表达细胞增殖标志基因：细胞增殖标志物（Ki67）基因,叉头盒M1蛋白（Foxm1）基因,增殖细胞核抗原（Pcna）基因和肝细胞生长因子（HGF）基因（P<0.05）;胆汁淤积组的原代肝细胞表达更低水平的肝细胞标志基因：白蛋白（Alb）基因,多药耐药相关蛋白2（Mrp2）基因,胆盐输出泵（Bsep）基因和肝细胞连环蛋白1（Catenin1）基因（P<0.05）,同时表达更高水平的胆管细胞标志基因：细胞角蛋白7（Ck7）基因,细胞角蛋白 19（Ck19）基因,胆管细胞多药耐药性蛋白1（Mdr1）基因和胆管细胞囊性纤维化跨膜传导调节因子（Cftr）基因（P<0.05）以及肝祖细胞标志基因：上皮细胞黏附分子（Epcam）基因和Y染色体性别决定区-盒转录因子9（Sox9）基因（P<0.05）。结论 胆汁淤积可诱导肝细胞向胆管细胞特性转化的可塑性。

Objective To explore the effect of bile duct ligation induced obstructive cholestasis on rat hepatocytes. Methods Ten Lewis rats were randomly divided into control group and cholestasis group, and the cholestasis was induced by bile duct ligation for 2 weeks. The histopathological changes were compared by H&E and aniline blue staining and the liver function was compared by biochemical analysis. Primary hepatocytes were isolated by modified two-step collagenase perfusion, and the expressions of hepatocyte marker genes, cell proliferation marker genes and cholangiocyte marker genes were detected by RT-qPCR. Results Compared with the control group,the liver of the cholestatic group showed obvious disordered histopathology, deposition of fibrous collagen and impaired liver function. Compared with the control group, the primary hepatocytes in the cholestasis group expressed higher cell proliferation-related genes（Ki67,Foxm1,Pcna and HGF）（P<0. 05）. Primary hepatocytes in the cholestasis group expressed lower levels of hepatocyte marker genes（Alb,Mrp2,Bsep and Catenin1）（P<0. 05）,and higher levels of cholangiocyte marker genes（Ck7,Ck19,Mdr1 and Cftr）（P<0. 05）and higher levels of the hepatic progenitor cell marker genes（Epcam and Sox9）（P<0. 05）. Conclusions Cholestasis induces rat hepatocyte plasticity in the transformation into bile duct properties.

       目的   评价血塞通联合右美托咪定对脑缺血再灌注损伤大鼠的脑保护效果。方法   选择老龄雄性Wistar大鼠50只，随机分为假手术（C）组、脑缺血再灌注（R）组、血塞通（P）组、右美托咪定（D）组，血塞通联合右美托咪定（PD）组，每组各10只。根据组别给予不同药物，行神经行为学测试；于第3、7天，测量脑梗死面积、脑水含量，以及超氧化物歧化酶（Superoxide dismutase，SOD）、谷胱甘肽过氧化酶（Glutathione peroxidase，GSH-PX）活性测定。结果   给药后第3、5、7天，与P、D组相比，PD组神经行为学评分改善更加显著（P＜0.001）；给药后第3、7天，与P组相比，PD组脑梗死面积、脑水含量均降低（P=0.01，P=0.002），SOD、GSH-PX活性升高显著（P=0.03，P=0.001）；与D组相比，PD组脑梗死面积、脑水含量也显著降低（P＜0.01，P=0.008）；SOD、GSH-PX活性升高显著（P=0.009，P＜0.001）。结论   血塞通联合右美托咪定较单独应用药物，能显著减轻缺血再灌注损伤造成的脑损害，具有脑保护作用。

       Objective  To explore the effects of Xuesaitong combined with dexmedetomidine on cerebral ischemia-reperfusion in elderly rats．Methods  Fifty elderly male Wistar rats were randomly divided into 5 groups：sham operation（C）group，cerebral ischemia-reperfusion（R）group，Xuesaitong（P）group，dexmedetomidine（D）group，Xuesaitong combined with dexmedetomidine（PD）group．Xuesaitong was given in group P，dexmedetomidine was given in group D，and normal saline was given in group C and group R，continuously for 7 days．After 3- and 7-day treatment，the brain of rats was dissected out to assay the area of cerebral infarction，degree of cerebral edema，superoxide dismutase（SOD） and glutathione peroxidase（GSH-PX） activity．Results  When compared PD group with P and D group，neurobehavioral score was lower at 3，5，7 day（P＜0.001）；area of cerebral infarction，degree of cerebral edema were less（P=0.01，P=0.002），activity of SOD and GSH-PX were higher at 3，7 days（P=0.03，P=0.001）respectively．When compared PD group with D group，area of cerebral infarction，degree of cerebral edema were less（P＜0.01，P=0.008），activity of SOD and GSH-PX were higher at 3，7 days（P=0.009，P＜0.001）respectively．Conclusions  The combination of Xuesaitong and dexmedetomidine can obviously reduce the damage by cerebral ischemia-reperfusion in elderly rats and has brain protective effects．