Immunoglobulin G from anti-glucose-6-phosphate isomerase antibodies positive patient with rheumatoid arthritis induces synovitis in cynomolgus monkeys

Anti-glucose-6-phosphate isomerase (GPI) antibodies (Abs) solely induce arthritis in mice. High titers of anti-GPI Abs are found in some patients with rheumatoid arthritis (RA), but their pathogenic role remains elusive. The aim of this study was to evaluate the pathogenic role of anti-GPI Abs in cynomolgus monkeys. IgG fractions were separated from sera of anti-GPI Abs-positive RA patients and healthy subjects and directly injected into the metacarpophalangeal joints of 4 cynomolgus monkeys. At day 16, the joints were harvested and examined histologically and immunohistochemically. The expression of C5a receptor (C5aR) molecule in the synovium was quantified by real-time PCR using cDNA from monkey joints. In monkey joints, IgG including anti-GPI Abs resulted in recruitment of granulocytes and mononuclear cells, strong deposition of human IgG on the articular surface, and overexpression of C5aR, but no joint swelling. No infiltrated cells or IgG deposition were observed in monkeys injected with IgGs from healthy subjects. Our results suggest that IgG fraction from RA patients including anti-GPI Abs may play a crucial role in the generation of synovitis in monkeys, although the pathogenesis of anti-GPI Abs in RA patients is still uncertain.     

Opa相互作用蛋白5在胰腺癌中的表达及其对PANC-1细胞增殖的影响

目的探索Opa相互作用蛋白5(OIP5)在胰腺癌中的表达及其对PANC-1细胞增殖的影响。方法通过数据库分析OIP5在胰腺癌组织及癌旁组织中的表达;用实时定量PCR(RT-qPCR)和蛋白印迹法(Western blot)分别检测人胰腺癌细胞系MIAPaCa-2、PANC-1、KP-3、BxPC-3细胞中OIP5 mRNA和蛋白表达;构建OIP5基因沉默质粒的慢病毒(pGCSIL-shOIP5)和对照质粒慢病毒(pGCSIL-shCtrl),分别感染PANC-1细胞,分为OIP5基因沉默组和shCtrl对照组,5 d后采用RT-qPCR和Western blot测定慢病毒敲低效率,流式细胞计量术检测细胞凋亡;OIP5基因沉默组和shCtrl对照组连续5 d进行MTT检测和细胞计数;OIP5基因沉默组和shCtrl对照组孵育10 d形成集落,Giemsa染色分别集落总数。结果胰腺癌中OIP5 mRNA表达显著高于正常胰腺组织(P<0.05),OIP5高表达患者的总存活率显著低于OIP5低表达患者(P<0.05),且其无病生存率也显著降低(P<0.05);OIP5在MIAPaCa-2、PANC-1和KP-3中表达较高,而在BxPC-3细胞系中的表达较低;MTT检测结果显示OIP5沉默在第4和第5天显著降低了PANC-1细胞的增殖速率(P<0.01);OIP5沉默后细胞集落数(平均为9个)显著低于shCtrl对照组中的数量(平均为40个)(P<0.01);OIP5沉默后PANC-1细胞凋亡比例为8.3%显著高于shCtrl的4.5%(P<0.01)。结论OIP5在胰腺癌细胞系中异常高表达,OIP5基因可调控胰腺癌PANC-1细胞的增殖、凋亡以及集落形成,提示OIP5可能在胰腺癌发病机制中作为癌基因发挥作用,从而为胰腺癌的靶向治疗提供了潜在的生物标志物。     

蛋白酶激活受体2激动剂对肥大细胞释放组胺的影响

目的 研究PAR-2激动剂(tc-LIGRLO-NH2与SLIGKV-NH2)和胰蛋白酶对结肠肥大细胞释放组胺的影响。方法 结肠组织经酶消化后,细胞成份用全HBSS重新悬浮。激发过程在LP4试管中、37℃条件下完成。组胺水平用以玻璃纤维为基础的荧光方法测定。结果PAR-2激动剂tc-LIGRLO-NH2和SLIGKV-NH2均可诱导人结肠肥大细胞剂量依赖性组胺释放。浓度为100μmol／mL时,tc-LIGRLO-NH2和SLIGKV-NH2可分别引起比基础分泌量多出2．5倍和2倍的组胺释放,而反PAR-2激动剂tc-OLRGIL-NH2和VKGILS-NH2在实验浓度高至300μmol／mL时仍对组胺释放无影响。胰蛋白酶在1．0～100μg/mL间可引起剂量相关性组胺释放,胰蛋白酶抑制剂可抑制之。PAR-2激动剂tc-LIGRLO-NH2的作用从加样后1min开始,3min后完成。细胞经过百日咳毒素和代谢抑制剂预先处理后,几乎失去了对tc-LIGRLO-NH2、SLIGKV-NH2和胰蛋白酶刺激的反应性。结论 PAR-2激动剂和胰蛋白酶是高效的组胺释放刺激剂,其在人结肠炎症性疾病中起一定的作用。     

MUS81基因在喉癌中的突变和表达

目的 探讨MUS81基因突变和表达与喉癌发生发展的相关性.方法 应用聚合酶链反应-单链构象多态性分析技术结合DNA测序检测分析了42例喉癌患者MUS81基因第9、10外显子的突变;应用半定量逆转录-PCR和Western印迹方法分析MUS81基因在喉癌组织中的表达情况.结果 42例喉癌、癌旁组织标本中,癌旁正常组织均无突变,喉癌组织标本中19例发生突变,占45.2%(19/42),11例喉癌组织第9外显子发现有突变,占26.2%(11/42),8 例喉癌组织第10外显子有突变,占19%(8/42),分析表明具有统计学意义(P<0.01).逆转录-PCR结果表明,42例喉癌中有17例MUS81基因mRNA低表达,占40.48%(17/42).Western印迹方法分析结果表明,42例喉癌中有17例MUS81蛋白质低表达,占40.48%(17/42),经统计学分析肿瘤组与对照组差异有统计学意义(P<0.01).分析表明MUS81基因突变与mRNA和蛋白质低表达有显著相关性(P<0.01).统计结果显示喉癌MUS81基因突变与TNM分期、年龄和淋巴结转移无相关性(P>0.05).MUS81基因低表达与TNM分期、年龄和淋巴结转移无相关性(P>0.05).结论 发现MUS81基因在喉癌组织中有突变发生及表达异常,提示MU81基因突变和表达异常可能是喉癌发生及发展的重要因素之一.     

耳穴静态电测量中的瞬变响应

耳穴电参数时变关系实验表明，在测量起始ｔ＜２τ时，因瞬变作用，电位Ｅ（ｔ）和压降Ｕ（ｔ）为瞬态响应，响应函数呈指数关系，特征参数为弛豫时间τ，τ≈ＲＣ；ｔ＞２τ时，为时变间期。电路分析给出数学描述，并与耳穴和模拟实验结果较相符。提示，时变特征应以ｔ＞２τ后提取，静态电测量时，采样应避开瞬变期，可提高准确性。该工作对正确鉴别时变性和特征提取，全面认识耳穴电特性具有重要意义。     

NF-κB哑铃形诱骗剂ODN对骨髓瘤细胞生长及IL-6表达的抑制作用

目的设计合成靶向NF-κB的哑铃形诱骗剂ODN,分析靶向NF-κB的哑铃形诱骗剂对NF-κB转录活性、多发性骨髓瘤细胞8266的生长及其分泌的IL-6的抑制效应。方法采用电泳迁移率变动分析(EMSA)体外检测诱骗剂ODN对NF-κB转录活性的抑制效应。将8266细胞随机分为传代培养的8266细胞组;诱骗剂ODN处理组及脂质体处理组。通过阳离子脂质体以2mg/L、4mg/L、8mg/L不同剂量诱骗剂ODN转染8266细胞。转染后8、12、18h,ELISA法检测8266细胞培养上清中IL-6的表达。用MTT比色检查诱骗剂ODN对IL-6刺激的8266细胞生长的影响。结果硫代磷酸的诱骗剂ODN在体外能有效地抑制NF-κB与其顺式元件的结合;2mg/L、4mg/L、8mg/L等不同浓度的脂质体-ODN复合物对8266细胞表达IL-6的抑制程度不同。脂质体-ODN复合物对8266细胞的生长及IL-6的活性均有抑制作用。结论靶向NF-κB的诱骗剂ODN在体外可抑制NF-κB的转录活性,从而抑制8266细胞的生长,降低瘤细胞中IL-6的表达。     

综合ICU中低年资护士培训方法探讨

目的探讨为综合ICU培养合格的护理人员的方法。方法根据ICU护士应具备的素质及低年资护士的特点,采用定期集中培训、一对一带教、参加护理查房等方法,对基础理论和基本技能进行强化,重点加强了ICU专科护理知识地训练。结果通过客观考核和主观考核,效果良好。结论经过为期12个月两个时段较规范的岗位培训,低年资护士能较好的适应ICU工作。     

Synergistic antimicrobial effect of cefotaxime and minocycline on proinflammatory cytokine levels in a murine model of Vibrio vulnificus infection.

BACKGROUND AND PURPOSE: Vibrio vulnificus causes primary bacteremia and necrotizing wound infection, leading to high morbidity and mortality in humans. This study aimed to evaluate the antimicrobial effect of cefotaxime and minocycline on proinflammatory cytokine levels in a murine model of V. vulnificus infection. METHODS: We investigated the dynamics of proinflammatory cytokines and their modulation by antimicrobial agents using a murine model of V. vulnificus infection. The change in cytokine levels was followed over a time course to identify the antimicrobial activity of the drugs against V. vulnificus. BALB/c female mice were challenged with an intraperitoneal infection using a clinical invasive isolate of Vv05191, and their cytokine levels were assayed over various time points. RESULTS: Serum levels of tumor necrosis factor-alpha, interleukin (IL)-1 beta, and IL-6 post-infection were found to be inoculum dose-dependent and positively correlated to the subsequent fatality rate in the infected mice. With an inoculum of 6.6 x 10(6) colony-forming units and intraperitoneal administration of cefotaxime, minocycline, or both, the serum and peritoneal fluid cytokine levels increased and then declined gradually. Comparison of the 3 antimicrobial regimens revealed that the magnitude of reduction in cytokine levels was greatest in mice treated with cefotaxime-minocycline combination. Moreover, the peritoneal fluid cytokine level in the combination group was significantly lower than that in the groups treated with minocycline or cefotaxime alone. CONCLUSIONS: The current results support the superiority of the combination therapy in treating invasive V. vulnificus infections.     

bFGF在骨组织工程学中的研究进展

碱性成纤维细胞生长因子 (b FGF)是近年来研究较为广泛的生长因子之一 ,对骨组织有着广泛的生物学效应。本文就 b FGF的研究进展及其目前在骨组织工程学研究方面的应用做一综述