Gadd45β和Gadd45γ在亚砷酸钠所致的MIHA细胞周期改变中的作用

目的 探索Gadd45β和Gadd45γ在亚砷酸钠所致的MIHA细胞周期改变中的作用,为砷中毒的人群防治提供依据。方法 收集贵州省兴仁县雨樟镇交乐病区砷中毒人群以及格沙屯正常人群分别作为砷中毒组和对照组,实时荧光定量PCR检测各组人群Gadd45β和Gadd45γ基因mRNA表达水平。同时,分别以不同剂量亚砷酸钠、不同时间处理MIHA细胞,流式细胞术测定细胞周期改变情况,实时荧光定量PCR和免疫印迹法分别检测Gadd45β和Gadd45γ基因mRNA表达水平和蛋白表达情况。在上述实验基础上,针对Gadd45β和Gadd45γ基因分别设计的siRNA作用于染砷的MIHA细胞,反向验证Gadd45β和Gadd45γ基因在砷致细胞周期改变过程中的影响。统计学分析采用独立样本t检验比较两组间差异,采用单因素方差比较多组间差异。结果 人群实验研究发现,与对照组相比,砷中毒患者Gadd45β和Gadd45γ基因mRNA表达水平升高（t = 2.576,P = 0.011;t = 2.312,P = 0.022）;MIHA细胞中,随亚砷酸钠染毒剂量、染毒时间增加,细胞G2/M期比例明显升高（F剂量 = 340.136,P＜0.001;F时间 = 49.194,P＜0.001）;在相对较低亚砷酸钠浓度（低于20 μmol/L）、一定时间内（低于48 h）Gadd45β和Gadd45γ基因mRNA和蛋白表达水平随染砷剂量、染砷时间升高而升高,超过该浓度范围和作用时间后,出现表达下降的现象（Gadd45β:F剂量-蛋白 = 37.568,P＜0.001;F剂量- mRNA = 9.771,P＜0.001;F时间-蛋白 = 61.144,P＜0.001;F时间- mRNA = 46.366,P = 0.001;Gadd45γ:F剂量-蛋白 = 12.989,P = 0.001;F剂量- mRNA = 23.613,P＜0.001;F时间-蛋白 = 27.425,P＜0.001;F时间- mRNA = 37.969,P＜0.001）。转染siRNA分别下调Gadd45β和Gadd45γ的表达后,细胞周期都出现G2/M期比例下调（t = 3.053,P = 0.038;t = 14.47,P＜0.001）。结论 砷致Gadd45β和Gadd45γ基因表达水平升高在其诱导MIHA细胞出现G2/M期阻滞过程中发挥重要作用。     

Berberine plays a cardioprotective role by inhibiting macrophage Wnt5a/β-catenin pathway in the myocardium of mice after myocardial infarction

Myocardial infarction (MI) is one of the diseases with high fatality rate. Berberine (BBR) is a monomer compound with various biological functions. And some studies have confirmed that BBR plays an important role in alleviating cardiomyocyte injury after MI. However, the specific mechanism is unclear. In this study, we induced a model of MI by ligation of the left anterior descending coronary artery and we surprisingly found that BBR significantly improved ventricular remodeling, with a minor inflammatory and oxidative stress injury, and stronger angiogenesis. Moreover, BBR inhibited the secretion of Wnt5a/β-catenin pathway in macrophages after MI, thus promoting the differentiation of macrophages into M2 type. In summary, BBR effectively improved cardiac function of mice after MI, and the potential protective mechanism was associated with the regulation of inflammatory responses and the inhibition of macrophage Wnt5a/β-catenin pathway in the infarcted heart tissues. Importantly, these findings supported BBR as an effective cardioprotective drug after MI.     

日本血吸虫尾蚴cDNA文库的免疫学筛选及EST序列的分析

目的为获得日本血吸虫（Schistosoma japonicura，Sj）转化生长因子β1（TGF—β1）基因的部分或全长cDNA序列。同时验证用针对某一蛋白的抗体筛选表达文库是否可以得到相应蛋白对应的基因序列。方法采用兔抗鼠TGF—β1血清，对Sj尾蚴cDNA表达文库进行免疫学筛选，对阳性克隆进行PCR扩增后，将大于500bp的克隆进行复筛，对复筛阳性的克隆进行测序和生物信息学鉴定。结果对大约10^6个噬菌斑进行了初筛。共获得7个阳性克隆；经过PCR扩增后，其中有4个克隆大于500bp，复筛获得3个持续阳性克隆；对测序后的阳性克隆进行生物信息学鉴定，得到的三个克隆均与日本血吸虫辅酶Q10氧化还原酶（SjCHGC）基因具有高的同源性（Bhata分值都大于200）。结论SjCHGC蛋白与兔抗鼠TGF—β1抗体的反应为交叉反应，用抗某一蛋白的抗体筛选表达文库得到相应蛋白的基因序列的方法不一定可行。     

头孢菌素类抗生素的耐药性比较研究

实验应用分光光度计观察了体外培养基中细菌在CPT,CTZ,β—内酰胺酶以及酶抑制剂MCIPC单独和相互作用下的生长浊度变化。①分别单用CPT(10MIC)和CTZ(15MIC)均可溶解细菌;而当加入微量酶液则可明显拮抗。②上述实验系统中同时加用MCIPC(1/16MIC)则能翻转酶对CPT和CTZ溶菌作用的拮抗而出现溶菌。③但当CPT或CTZ分别与酶在菌液中孵育10分或30分钟后,再加用等量MCIPC时仅能翻转酶对第三代头孢菌素-CTZ的拮抗,重新再现溶菌作用,而此时对第一代头孢菌素—CPT的溶菌作用则不能再现。该实验为第三代头孢菌素的非水解屏障耐药机制提供了重要依据。     

β-环连蛋白及其mRNA在肾癌中的表达

目的　探讨β -环连蛋白在肾癌发生发展过程中的作用及其变化机制。方法　我们采用免疫组织化学、Westernblot、RT -PCR等方法对本院收治的 2 6例肾癌患者手术标本进行了研究。结果　 2 6例患者中有 2 5例癌组织中细胞胞浆内β 环连蛋白表达明显高于正常组织内 ,而且肿瘤分期越高 ,癌细胞内β 环连蛋白的表达也越强 ,pT3和pT4期的肿瘤内的表达明显高于pT1和pT2期的肿瘤 ,P <0 .0 1,而其β -环连蛋白mRNA的表达水平并无明显变化。结论　细胞内 β -环连蛋白表达的增加与肾癌的发生发展有关 ,而且可能是肾癌形成和发展的重要机制之一。     

孕妇血清标记物在中孕筛查中的应用

目的 :通过对孕妇血清标记物甲胎蛋白 (AFP)和人绒毛膜促性腺激素 ( β -HCG )测定来筛选缺陷儿。 方法 :用放射免疫法对 6 2例孕 14～ 2 3周妊娠妇女上述标记物连测 3次 ,每次隔 12～ 14d ,以两次异常为高危个体。每例筛查者跟踪至胎儿出生后。结果 :在筛查中发现了 1例先天愚型和 1例无脑儿。结论 :上述两种血清标记物可以作为神经管缺陷和先天愚型筛查的一项常规方法     

黄芩甙、地塞米松对大鼠感染性脑水肿细胞因子的影响

目的 :探讨黄芩甙、地塞米松对大鼠感染性脑水肿白细胞介素 1(IL 1β)、肿瘤坏死因子α(TNF α)的影响。方法 :应用大鼠感染性脑水肿模型 ,采用干湿法、原子吸收分光光度法、ELISA测定各组脑组织水含量、钠离子含量及脑匀浆上清液中IL 1β,TNF α水平的变化 ;在光镜下观察各组脑组织的细胞形态学改变。结果 :在大鼠感染性脑水肿(感脑 )组脑水含量、钠离子、IL 1β、TNF α含量均比生理盐水 (对照 )组明显增高 (P <0 .0 1)。在黄芩甙和地塞米松两组则上述指标均下降 ,低于感脑组 (P <0 .0 5 ) ;而两组间各指标比较差异无显著性 (P >0 .0 5 ) ;脑组织水含量与IL 1β ,TNF α含量成正相关 (r分别为 0 9381,0 8349,P <0 .0 1)。对照组脑组织结构正常 ,感脑组织脑组织可见弥漫的灶性水肿区 ;黄芩甙和地塞米松组水肿明显减轻。结论 :黄芩甙、地塞米松对大鼠感染性脑水肿有保护作用 ,效果相近 ;作用机制与抑制IL 1β ,TNF α过度产生有关     

Pathogenetic role and clinical significance of interleukin-1β in cancer

In recent years, pro-oncogenic mechanisms of the tumour microenvironment (ТМЕ) have been actively discussed. One of the main cytokines of the TМЕ is interleukin-1 beta (IL-1β), which exhibits proinflammatory properties. Some studies have shown an association between an increase in IL-1β levels and tumour progression. The purpose of this review is to analyse the pathogenic mechanisms induced by IL-1β in the TМЕ, as well as the diagnostic significance of the presence of IL-1β in patients with cancer and the efficacy of treatment with IL-1β inhibitors. According to the literature, IL-1β can induce an increase in tumour angiogenesis due to its effects on the differentiation of epithelial cells, pro-angiogenic molecule secretion and expression of adhesion molecules, thus increasing tumour growth and metastasis. IL-1β is also involved in the suppression of anti-tumour immune responses. The expression and secretion of IL-1β has been noted in various types of tumours. In some clinical studies, an elevated level of IL-1β was found to be associated with low efficacy of anti-cancer therapy and a poor prognosis. In most experimental and clinical studies, the use of IL-1β inhibitors contributed to a decrease in tumour mass and an increase in the response to anti-tumour drugs.