Ang-(1-7)与肺高压状态下肺血管平滑肌细胞的增生*☆

背景：Ang-(1-7)虽然具有抗血管平滑肌细胞增殖作用,但在不同的血管床中其作用可能存在差异。直接给予外源性Ang-(1-7)是否可抑制肺动脉高压大鼠肺血管平滑肌细胞的增殖尚不清楚。 目的：探讨Ang-(1-7)对野百合碱诱导的肺动脉高压大鼠肺血管平滑肌细胞增殖的影响。 方法：雄性SD大鼠颈部一次性注射60 mg/kg野百合碱制备肺动脉高压模型。24 h后,分别经微泵持续泵入Ang-(1-7)(治疗组)或生理盐水(模型组),并设立未造模的对照组。给药2,4周,测定大鼠的右心室收缩压、心室质量、肺小动脉管壁厚度占管径的百分比及管壁面积占血管总面积的百分比。免疫组织化学方法检测肺血管平滑肌细胞α-平滑肌肌动蛋白及增殖细胞核抗原的表达。 结果与结论：野百合碱诱导2周,与对照组比较,模型组大鼠右心室收缩压、各心室的质量无明显变化,肺小动脉管壁厚度占管径的百分比、管壁面积占血管总面积的百分比、增殖细胞核抗原阳性率显著增高,α-平滑肌肌动蛋白显著降低;野百合碱诱导4周,模型组大鼠右心室收缩压、各心室的质量、肺小动脉管壁厚度占管径的百分比、管壁面积占血管总面积的百分比、增殖细胞核抗原阳性率均显著增高,α-平滑肌肌动蛋白显著降低。而治疗组上述指标与对照组比较差异无显著性意义 (P > 0.05)。说明在野百合碱诱导的肺动脉高压模型中,在肺动脉压增高之前已有肺血管形态学的变化,Ang-(1-7)可通过减轻肺血管平滑肌细胞的增生抑制大鼠肺动脉压的升高。     

大鼠细胞因子信号转导抑制因子-1基因的多态性及其结构分析

目的 扩增大鼠细胞因子信号转导抑制因子-1(SOCS-1)基因,利用生物信息学方法预测其结构和功能特征,为进一步的实验研究提供理论指导.方法 扩增并测序大鼠的SOCS-1编码区序列,利用生物信息学网站的在线分析工具和Vector NTI suite软件包,识别大鼠SOCS-1基因并预测其编码蛋白质的各种结构特征,根据该基因构建其分子进化树.结果 聚合酶链反应(PCR)扩增获得2个序列不同的SOCS-1基因,BLASTx分析该基因为全长基因,该基因全长639 bp,编码212个氨基酸(aa),具有1个SOCS盒(aa172～aa208),1个Scr同源区2(SH2)结构域(aa80～aa155)和1个核定位序列(aa160～aa174),一级结构含有2个线性B细胞抗原表位,全部位于蛋白的表面,并在空间结构上相距较远.结论 SOCS-1基因具有基因多态性,其保守的SH2区域及SOCS盒与其对信号转导通路抑制作用有关,而其核定位序列可能影响其他核转导因子,B细胞线性表位则在免疫诊断方面有较好的应用前景.     

慢性血吸虫感染对脓毒症小鼠保护作用的研究

目的 初步探讨慢性血吸虫(SJ)感染对脓毒症小鼠的保护作用及其机制.方法 选择BALB/c雄性小鼠,按随机数字表法分组进行三部分实验.实验1:经腹部皮肤接种SJ尾蚴感染8周建立慢性SJ感染模型,分为正常组和SJ组,每组10只;用酶联免疫吸附法(ELISA)检测血清白细胞介素(IL-4和IL-10)、肿瘤坏死因子-α(TNF-α)、γ-干扰素(IFN-γ)水平,实时荧光定量聚合酶链反应(PCR)检测腹腔巨噬细胞IL-10和TNF-α的mRNA表达,了解慢性SJ感染小鼠免疫状态.实验2:以脂多糖(LPS)腹腔注射诱导小鼠脓毒症模型,分为LPS组和SJ-LPS组,每组15只;用ELISA法动态观察注射LPS后0、24、48和72 h细胞因子的变化,0 h的水平相当于正常小鼠和SJ感染8周水平,观察慢性SJ感染对脓毒症过程的影响.实验3:分别以盲肠结扎穿孔术(CLP)和LPS诱导两种不同的脓毒症模型,评价慢性SJ感染对脓毒症小鼠72 h存活率的影响.结果 实验1:SJ组血清抗炎因子IL-4[(151.35±12.24)ng/L]和IL-10[(133.22±11.09)ng/L]水平较正常组[IL-4(56.32±8.66)ng/L,IL-10(48.17±7.23)ng/L]显著升高(均P＜0.05),并可使巨噬细胞向替代活化性巨噬细胞分化,慢性SJ感染使腹腔巨噬细胞高表达IL-10 mRNA(SJ组4.46±1.82,正常组1.52±0.60),抑制TNF-α mRNA表达(SJ组1.61±0.93,正常组2.32±1.03,均P＜0.05).实验2、3:慢性SJ感染小鼠血清IL-4、IL-10于注射LPS后0 h即显著升高,随后下降,至72 h仍明显高于LPS组[IL-4(ng/L):92.2±7.6比41.5±4.5;IL-10(ng/L):92.1±7.8比35.6±4.0,均P＜0.05];TNF-α、IFN-γ均于24 h达峰值后逐渐下降,至72 h SJ-LPS组仍显著低于LPS组[TNF-α(ng/L):82.9±5.6比91.5±5.2;IFN-γ(ng/L):44.1±4.8比52.6±4.0,均P＜0.05].慢性SJ感染可明显改善CLP或LPS所致脓毒症小鼠的存活率(CLP:80%比20%,LPS:70%比30%,均P＜0.05).结论 慢性SJ感染可使脓毒症小鼠血清抗炎因子升高,存活率上升,从而起到保护作用.

Abstract：

Objective To preliminarily study the protective effect of chronic schistosoma japonica (SJ)infestation against sepsis in mice and its mechanism. Methods BALB/c male mice were used, and the experiment was divided into three parts. Experiment 1: chronic SJ infestation model was reproduced by SJ cercaria inoculation through abdominal skin for 8 weeks. Twenty mice were randomly grouped into normal group (n=10) and SJ group (n=10). The levels of interleukins (IL-4, IL-10), tumor necrosis factor-α(TNF-α) and interferon-γ (IFN-γ) in serum were detected by enzyme linked immunosorbent assay (ELISA).Real-time polymerase chain reaction (PCR) was employed to detect the levels of IL-10 mRNA and TNF-αmRNA in abdominal macrophages. This experiment was meant to evaluate immune state in mice with chronic SJ infestation. Experiment 2: lipopolysaccharide (LPS) was intraperitoneally injected to reproduce sepsis model. Thirty mice were randomly grouped into LPS group (n=15) and SJ-LPS group (n=15). The levels of cytokines were determined by ELISA at 0, 24, 48 and 72 hours after LPS injection. This experiment was meant to detect the effect of chronic SJ infestation in mice during the septic process. Experiment 3 : two types of sepsis model were reproduced by cecal ligation and puncture (CLP) and LPS injection, respectively. The survival rate of mice with chronic SJ infestation in 72 hours in either type of sepsis was evaluated. Results Experiment 1, compared with normal group [IL-4 (56.32±8.66) ng/L, IL-10 (48.17±7.23) ng/L],chronic SJ infestation showed an increase in serum IL-4 [(151. 35 ± 12. 24) ng/L] and IL-10 [(133. 22 ±11. 09) ng/L, both P＜0. 05]. Chronic SJ infestation also resulted in an increase in IL-10 mRNA expression (SJ group 4. 46±1. 82, normal group 1. 52±0. 60) and inhibited TNF-α mRNA expression (SJ group 1. 61±0.93, normal group 2. 32±1.03) in abdominal macrophages (both P＜0. 05), indicating that macrophages could be differentiated into alternative activated macrophages. Experiments 2 and 3 showed that the levels of serum IL-4 and IL-10 were increased at 0 hour after LPS injection, and then gradually decreased in SJ-LPS group, but the levels were still higher than those in LPS group at 72 hours [IL-4 (ng/L): 92. 2±7. 6 vs.41.5±4. 5; IL-10 (ng/L): 92. 1±7. 8 vs. 35. 6±4. 0, both P＜0. 05]; the levels of TNF-α and IFN-γ were increased at 24 hours, and then decreased in SJ-LPS group, and the levels were lower than those in LPSgroup at 72 hours [TNF-α (ng/L): 82. 9±5. 6 vs. 91. 5±5. 2; IFN-γ (ng/L): 44.1±4. 8 vs. 52. 6±4. 0,both P＜0. 05]. Therefore, chronic SJ infestation could improve the survival rate of mice with sepsis induced by CLP or LPS (CLP: 80% vs. 20%, LPS: 70% vs. 30%, both P＜0.05). Conclusion Chronic SJ infestation could elevate anti-inflammatory factors in septic mice, thus ameliorating the survival rate, so it has protective effect on mice with sepsis.     

同时对首次血液透析患者和家属进行心理干预对维持性血透患者焦虑状态的影响

目的：探讨同时对首次血液透析患者和家属进行心理干预对维持性血透患者焦虑状态的影响。方法：将120例进行首次血液透析的终末期肾病患者120例随机分为试验组（60例）和对照组（60例）,对照组只对患者进行宣教和心理干预,试验组则同时对患者及其家属进行针对性的心理干预,观察两组患者SAS评分的变化情况。结果：干预后,两组患者的SAS评分均显著降低（P〈0.01）,且试验组显著低于对照组（P〈0.01）。结论：同时对首次血液透析患者和家属进行心理干预可显著降低维持性血透患者的焦虑水平。     

张掖市＜5岁儿童乙型肝炎病毒携带率和抗体水平监测结果分析

[目的]了解张掖市＜5岁儿童乙型病毒性肝炎(乙肝)感染现状和免疫抗体水平. [方法]采用分层抽样和单纯随机抽样的方法,采集1 178名0～4岁儿童血清标本,检测乙肝病毒表面抗原(HBsAg)、乙肝病毒表面抗体(抗-HBs)阳性率水平. [结果]HBsAg阳性率1.19%,不同年龄组儿童HBsAg阳性率差异有统计学意义,不同县(区)HBsAg阳性率差异无统计学意义,农村儿童HBsAg阳性率高于城镇儿童;抗-HBs阳性率66.55%,不同年龄组、不同县(区)儿童抗-HBs阳性率差异均有统计学意义,农村儿童抗-HBs阳性率低于城镇儿童. [结论]国际疫苗免疫联盟(GAVI)项目乙肝疫苗(HepB)接种达到了预期的免疫效果.今后HepB免疫重点人群是农村地区儿童、母亲HBsAg阳性儿童;有必要对4岁以上儿童开展乙肝疫苗(HepB)加强免疫.     

慢性血吸虫感染对脓毒症小鼠保护作用的研究

Objective To preliminarily study the protective effect of chronic schistosoma japonica (SJ)infestation against sepsis in mice and its mechanism. Methods BALB/c male mice were used, and the experiment was divided into three parts. Experiment 1: chronic SJ infestation model was reproduced by SJ cercaria inoculation through abdominal skin for 8 weeks. Twenty mice were randomly grouped into normal group (n=10) and SJ group (n=10). The levels of interleukins (IL-4, IL-10), tumor necrosis factor-α(TNF-α) and interferon-γ (IFN-γ) in serum were detected by enzyme linked immunosorbent assay (ELISA).Real-time polymerase chain reaction (PCR) was employed to detect the levels of IL-10 mRNA and TNF-αmRNA in abdominal macrophages. This experiment was meant to evaluate immune state in mice with chronic SJ infestation. Experiment 2: lipopolysaccharide (LPS) was intraperitoneally injected to reproduce sepsis model. Thirty mice were randomly grouped into LPS group (n=15) and SJ-LPS group (n=15). The levels of cytokines were determined by ELISA at 0, 24, 48 and 72 hours after LPS injection. This experiment was meant to detect the effect of chronic SJ infestation in mice during the septic process. Experiment 3 : two types of sepsis model were reproduced by cecal ligation and puncture (CLP) and LPS injection, respectively. The survival rate of mice with chronic SJ infestation in 72 hours in either type of sepsis was evaluated. Results Experiment 1, compared with normal group [IL-4 (56.32±8.66) ng/L, IL-10 (48.17±7.23) ng/L],chronic SJ infestation showed an increase in serum IL-4 [(151. 35 ± 12. 24) ng/L] and IL-10 [(133. 22 ±11. 09) ng/L, both P＜0. 05]. Chronic SJ infestation also resulted in an increase in IL-10 mRNA expression (SJ group 4. 46±1. 82, normal group 1. 52±0. 60) and inhibited TNF-α mRNA expression (SJ group 1. 61±0.93, normal group 2. 32±1.03) in abdominal macrophages (both P＜0. 05), indicating that macrophages could be differentiated into alternative activated macrophages. Experiments 2 and 3 showed that the levels of serum IL-4 and IL-10 were increased at 0 hour after LPS injection, and then gradually decreased in SJ-LPS group, but the levels were still higher than those in LPS group at 72 hours [IL-4 (ng/L): 92. 2±7. 6 vs.41.5±4. 5; IL-10 (ng/L): 92. 1±7. 8 vs. 35. 6±4. 0, both P＜0. 05]; the levels of TNF-α and IFN-γ were increased at 24 hours, and then decreased in SJ-LPS group, and the levels were lower than those in LPSgroup at 72 hours [TNF-α (ng/L): 82. 9±5. 6 vs. 91. 5±5. 2; IFN-γ (ng/L): 44.1±4. 8 vs. 52. 6±4. 0,both P＜0. 05]. Therefore, chronic SJ infestation could improve the survival rate of mice with sepsis induced by CLP or LPS (CLP: 80% vs. 20%, LPS: 70% vs. 30%, both P＜0.05). Conclusion Chronic SJ infestation could elevate anti-inflammatory factors in septic mice, thus ameliorating the survival rate, so it has protective effect on mice with sepsis.     

可溶性血栓调节蛋白与冠心病患者短期预后的关系

[目的]检测冠心病患者可溶性血栓调节蛋白(soluble thrombomodulin,sTM)水平并探讨其临床意义.[方法]检测冠心病患者入院后24 h内和出院前24 h血浆sTM水平,并通过记录住院期间主要心血管事件和随访出院后6个月内严重心血管事件的发生情况,研究冠心病患者循环血浆sTM的变化及与患者预后间的关系.[结果]住院期间主要心脏事件发生组sTM水平高于主要心脏事件未发生组,随访期间严重心血管事件发生组血浆sTM较严重心血管事件未发生组水平增高.[结论]入院时的sTM水平与住院期间的心血管事件有密切的相关性,出院时的sTM与患者出院后的心脏事件关系密切,是冠心病患者短期预后的强有力的预测因子.     

大鼠移植静脉再狭窄过程中黏着斑激酶的活化及奥美沙坦的干预作用

目的研究黏着斑激酶(FAK)在大鼠移植静脉再狭窄过程中的作用,并研究奥美沙坦的干预效应。方法建立大鼠颈外静脉移植模型,将40只雄性大鼠随机分成:1)假手术组;2)模型组;3)奥美沙坦组;4)生理盐水组等4组。观察各组血管壁内膜厚度及内膜/中膜(I/M);采用免疫组化方法观察各组血管平滑肌细胞增殖核抗原(PCNA)及平滑肌肌动蛋白(α-SM actin)表达;采用 western-blot 方法检测 FAK 及磷酸化 FAK 的表达。结果模型组与假手术组相比,内膜明显增厚(P<0.01),I/M 明显增加(P<0.01),PCNA 表达明显增加(P<0.01),α-SMactin、FAK、磷酸化 FAK 叫显增加(P<0.05);奥美沙坦组与模型组相比,内膜厚度、I/M 明显减轻(P<0.05),PC-NA、α-SM actin、磷酸化 FAK 表达明显降低(P<0.05)。结论 FAK 活化参与了大鼠移植静脉再狭窄过程,奥美沙坦可以抑制大鼠移植静脉再狭窄,这种作用可能与减轻局部 FAK 活化有关。     

动态三维超声心动图诊断左心瓣膜疾病的价值研究

【目的】探讨动态三维超声心动图诊断左心系统瓣膜疾病的临床应用价值。【方法】应用动态三维（3D）重建技术研究60例有左心系统瓣膜疾病的患者。全部病例经胸壁二维超声（TTE）、经食道超声（TEE）检查及3D重建，形成动态三维超声心动图，观察瓣膜的形态、结构及活动情况。对49例行手术患者观察术中瓣膜情况并与二维及3D图像进行对比。对行主动脉瓣置换术者将TTE、TEE及3D状态下的主动脉瓣环最大径测值与人工瓣之间进行线性相关分析并建立回归方程。【结果】3D重建能显示瓣膜的整体结构；3D所诊断的瓣膜局部解剖信息全部被手术所证实，在部分病变的显示中，3D超声能补充纠正TTE的诊断：TEE、3D的主动脉瓣环最大径测值与人工瓣大小有良好的相关性（P〈0．05），较TTE更为密切（P〈0．05）。【结论】动态三维超声心动图像较经胸二维超声全面清晰，可为左心瓣膜病变提供更准确的诊断信息；动态三维超声能较准确地测量主动脉瓣环最大径，在换瓣手术中能为人工瓣大小的选择提供参考。     

奥美沙坦改善大鼠移植静脉内膜重构及其超声评价

目的:观察大鼠颈外静脉移植后的内膜重构变化,研究奥美沙坦对内膜重构的干预作用。方法:制作大鼠颈外静脉移植模型,48只雄性大鼠被随机分成:①假手术组;②模型对照组;③奥美沙坦组;④生理盐水对照组等4组。观察各组血管壁内膜厚度,采用血管超声方法,观察大鼠颈外静脉移植后的血管超声显像,观察奥美沙坦干预前后各组血管搏动指数(pulsatility index,PI)的变化。结果:与假手术组相比,模型对照组移植静脉内膜厚度明显增加(P<0.01),PI明显增加(P<0.01);与模型对照组相比,奥美沙坦组桥静脉内膜厚度明显减轻(P<0.05),PI明显减低(P<0.05),生理盐水对照组与模型对照组相比内膜厚度无显著变化(P>0.05),PI值无显著变化(P>0.05)。超声检查可清晰显示移植静脉血管的走行、血管腔径大小,吻合口的延续情况。结论:奥美沙坦可以减轻大鼠移植静脉内膜重构,并改善移植血管的弹性功能,超声检查可以评估移植静脉内膜重构情况。