目的 探讨丙酸睾酮注射液联合乌司他丁对脓毒症患者免疫失衡的调节作用。方法 选取我院2019年10月—2020年1月收治的88例脓毒症患者,随机分成观察组和对照组各44例,对照组采用乌司他丁配合常规治疗,观察组在此基础上联合丙酸睾酮注射液对患者治疗,比较治疗1周2组临床疗效差异,观察治疗前及治疗1周,2组炎症因子水平[白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-1β(IL-1β)]、睾酮(T)、白蛋白(Alb)水平及疾病危重程度(APACHEⅡ评分)、器官衰竭程度(SOFA评分)变化,分析治疗1周内2组患者药物不良反应发生情况差异。结果 治疗1周,观察组总有效率高于对照组(P＜0.05);2组患者IL-6、IL-1β水平及APACHEⅡ、SOFA评分均降低,且观察组较对照组更低(P＜0.05);2组患者IL-10、T、Alb水平均升高,且观察组较对照组更高(P＜0.05);2组药物不良反应总发生率比较无统计学意义(P>0.05)。结论 丙酸睾酮注射液联合乌司他丁治疗脓毒症可取得良好临床疗效,可有效改善病情及预后,对患者免疫失衡调节有积极意义。

Objective To investigate the regulatory effect of testosterone propionate injection combined with ulinastatin on immune imbalance in patients with sepsis.Methods A total of 88 patients with sepsis treated in our hospital from October 2019 to January 2020 were randomly divided into observation group and control group,with 44 cases each.The control group was treated with ulinastatin combined with routine treatment,while the observation group was treated with testosterone propionate injection additionally.The clinical efficacy difference between the two groups were compared after treating 1 week.The levels of inflammatory factors [interleukin-6 (IL-6),interleukin-10 (IL-10),interleukin-1 β (IL-1β)],testosterone (T),albumin (Alb),the severity of disease (APACHE II score) and the degree of organ failure (SOFA score) in the two groups were observed before and 1 week after treatment.The differences of adverse drug reactions between the two groups within 1 week of treatment were analyzed.Results The total effective rate of the observation group was higher than that of the control group (P＜0.05).IL-6 and IL-1β levels and APACHE II and SOFA scores decreased after treatment,while those of the observation group were lower than the control group (P＜0.05).The levels of IL-10,T and Alb in the two groups were increased,while those in the observation group were higher than the control group (P ＜ 0.05); the total incidence of adverse drug reactions in the two groups was not statistically significant (P>0.05).Conclusions Testosterone propionate injection combined with ulinastatin had good clinical efficacy in the treatment of sepsis,effectively improved the condition and prognosis,and had positive significance in the regulation of immune imbalance.

目的 探索内源性神经干细胞在大鼠海马可溶性因子中的体外发育归宿及分化鉴定。方法 显微镜下分离Wistar大鼠海马组织放置于低温DMEM/12培养基,低温振荡2小时后高速离心(15000 g),获取实验所用海马组织可溶性因子。取材出生1天的Wistar乳鼠海马中的内源性神经干细胞(endogenous neural stem cells, ENSCs),将ENSCs分别于含海马可溶性因子终浓度为0(对照组)、50、100、200、400 μl/mL的无血清DMEM/F12培养基中培养6天并每日观察,使用免疫细胞化学、Western Blot印记技术比较各组ENSCs中Nestin、CD133的表达量;同时计量并比较各组ENSCs成球个数,以探索在模拟颅内微环境情况下,ENSCs发育、归宿及分化。进一步于最适宜的海马可溶性因子终浓度中分化神经球,对分化的细胞行神经元特异性蛋白入(如:β-tubullin III、MAP2)及胶质细胞特异性蛋白(如:GFAP、S100及p75 NGFR)免疫细胞化学检测。结果 大鼠ENSCs在培养基中呈单细胞漂浮生长,球形; ENSCs于海马可溶性因子各实验分组中培养第2天呈细胞球状态,对照组中无细胞球形成(与100 μl/mL组比较,P₁=0.00),100 μl/mL组与对照组比较有统计学意义(P₁=0.00<0.05);至第6天,在100 μl/mL组中的细胞球数量明显多于其余各组(P₁'=P₂'=P₃'=P₄'=0.00)。在免疫细胞化学检测中,100 μl/mL组中细胞球表达干细胞高亲和蛋白Nestin、CD133阳性,Western Blot免疫印迹检测其中Nestin、CD133蛋白高于对照组。进一步分化试验中,细胞球呈贴壁生长的单细胞状态、有突起伸出、长梭形,免疫细胞化学检测分化的细胞表达胶质细胞特异性蛋白GFAP、S100、p75NGFR阳性,但不表达神经元特异性蛋白β-tubullin III与MAP2。结论 大鼠ENSCs在终浓度为100 μl/mL的HSF作用下,可促进 ENSCs的增殖分裂;ENSCs在同样浓度下的HSF中可进一步分化为表达GFAP、S100、p75NGFR阳性的胶质样细胞;100 μl/mL的HSFS是ENSCs的一种生理性诱导剂或参与促进ENSCs增殖、分化及通过细胞替代或因子分泌等机制修复神经损伤。

Objective The aim of this study was to explore induction and differentiation of endogenous neural stem cells(ENSCs) in the hippocampus soluble factors(HSF) from the hippocampus of adult Wistar rats by mimicking an intracranial microenvironment. Methods After Wistar rats sacrificed, the hippocampus tissue was obtained in cold DMEM/F12. After centrigued and filtered, the HSF was stored at -20℃. The ENSCs was obtained from the hippocampus tissue of a neonate Wistar rat. Collected the tissue, digested and obtained the ENSCs. After we observed the morphology, the ENSCs were cultured in different concentration (0、50、100、200、400 μl/mL) of HSF for 6 days, and compared the expression of Nestin and CD133 by immunocytochemistry. Meanwhile,we compared the Nestin and CD133 protein by western blot. And then we explored the optimal concentration of HSF by the numbers of all groups on the second and sixth day. Furthermore, we did the differentiated experiment using the same concentration of HSF. Results The number of neurospheres in the 100 μg/mL group was significantly higher than those in the other groups on the 6th day. Immunofluorescence revealed that the neurospheres from ENSCs in the 100 μg/mL group more highly expressed nestin and CD133 than control. This result was confirmed by western blot analysis. The neurospheres can differentiate into glia-like cells in 100 μg/mL HSF and 1% FBS expressing GFAP, S100 and P75 NGFR by immunofluorescence. Conclusion These data indicated that HSF alone, mimicking a destination of ENSCs in vitro, could induce and differentiate neurospheres from ENSCs, as a new method to get NSCs and glia-like cells differentiated from ENCs to repair the diseases of center nervous system.

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.