SD大鼠体外受精与早期胚胎体外培养体系的初步建立

目的改造IVF-30和HTFplus培养液,观察在改造后培养液里卵的受精和早期胚胎的发育状况,寻找适合SD大鼠体外受精和早期胚胎培养的实验体系。方法采用两种方法(超数排卵及体内成熟)采集成熟卵母细胞用于体外受精。选择两种商品化的培养液IVF-30(Vitrolife),HTFplus(Lifeglobal)。分别在以上的培养基中增加10、20、30mmol/L的NaCl,将培养液的渗透压提升至325~355mOsM之间,观察在改造前与改造后的培养液内SD大鼠卵母细胞受精率和囊胚发育率等指标。结果经改造的培养液HTFplus和IVF-30,体外受精率分别从13%、15%提升至70%、67%。改造后的IVF-30和HTFplus的2细胞,大于4细胞及囊胚的发育率分别为85%,74%,22%;96%,75%,32%。HTFplus,IVF-30经过改造后更适合于SD大鼠的体外受精及早期胚胎培养。与超排相比较更多的在自然周期的体内成熟卵母细胞(16%,28%)发育到了囊胚。结论成功建立了SD大鼠的体外受精和体外培养系统。     

聚乙烯吡咯烷酮K30在甲硝唑片制备中的应用

目的研究辅料聚乙烯吡咯烷酮K30(PVPK30)对甲硝唑片质量的影响.方法依照中国药典2000年版二部测定甲硝唑片的溶出度.结果加入辅料PVPK30的甲硝唑片较原处方的溶出度明显加快,两者T50有显著差异(片1T50=11.5 min,片2T50=6.8 min).结论辅料PVPK30能提高甲硝唑片的质量.     

间变性大细胞淋巴瘤1例报告

介绍诊治的1例间变性大细胞淋巴瘤的临床特征、形态学、免疫表型及遗传学特征,并复习相关文献.     

脑星形细胞瘤中TIP30、VEGF和CD34的表达及其相关性研究

Objective To investigate the expression and relationship of TIP30(HIV-1 Tat interactive protein 2), vascular endothelial growth factor (VEGF) and MVD (detected by CD34) in the angiogenesis of human brain astrocytomas. Methods Expression of TIP30, VEGF and CD34 in 19 cases of normal brain tissue and 71 cases of astrocytoma were immunohistochemically examined with Elivision plus two-step method. Results The positive expression of TIP30 could be seen in cytoplasm of neuroglial cells and neurons of 19 normal brain tissues. The positive expression rate of TIP30 in 71 cases of astrocytoma was 33.80 % (24/ 71). The positive expression rate of TIP30 in astrocytoma of different grades was 52 % for grade Ⅱ, 34.78 % for grade Ⅲ and 13.04 % for grade Ⅳ. The positive expression rate of TIP30 in high grade (Ⅲ+Ⅳ) of astrocytoma was found significantly lower than that in low grade(Ⅱ) (χ2=5.71, P <0.05); The expression of VEGF and MVD detected by CD34 in astrocytomas were higher than that in normal brain tissue and increased as the tumor grade increased; In astrocytoma, the negtive correlation was found between the expression of TIP30 and VEGF (r=-0.428, P<0.05); no correlation was found between TIP30 and MVD(r=-0.065, P 0.05); the positive correlation was found between VEGF and MVD(r=0.684, P<0.01). Conclusion The positive expression rate of TIP30 in normal brain tissue is significantly higher than that in astrocytoma. The positive expression rate of TIP30 significantly decreases as the pathological grade of the astrocytoma increases; The expression of TIP30 and VEGF is negatively correlated in astrocytoma.     

浙江省局部地区2002～2003年流行的无菌性脑膜脑炎的病原学检测与分析

浙江省局部地区 2 0 0 2～ 2 0 0 3年多次出现无菌性脑膜脑炎的流行。为对疫情进行病原学检测与分析 ,采集了临床标本 ,采用HEp 2、RD、Vero等细胞分离病毒 ,并用肠道病毒通用引物进行特异性逆转录 聚合酶链反应 (RT PCR)扩增。结果显示 :5 1份脑脊液标本中分离出埃可病毒 30型 (ECHO3 0 ) 5株 ,柯萨奇病毒B组 5型 (Coxsack ievirusB5,Cox B5) 1株 ;6 8份粪便标本中分离出ECHO3 0 34株 ,Cox B51株。RT PCR的结果与上述病毒分离结果相一致。此外 ,对采集的 1 5例患者的急性期、恢复期双份血清测定了特异性中和抗体 ,其中 1 3例患者抗体呈≥ 4倍增长。证实引起浙江省局部地区 2 0 0 2～ 2 0 0 3年无菌性脑膜脑炎的主要病原为ECHO3 0 。     

MiR-30a-5p通过泛素水解酶22抑制人宫颈癌细胞上皮-间质转化功能

【目的】研究microRNA-30a-5p（miR-30a-5p）对人宫颈癌Hela细胞上皮-间质转化功能的影响及其相关机制。【方法】宫颈癌Hela细胞株分别转染目的mir的模拟物和阴性对照模拟物,分别以30a-5p组、NC组命名并标记细胞。同时,以未经过处理的Hela细胞作为对照（Control组）。分别用逆转录-聚合酶链反应法检测各组宫颈癌细胞的miR-30a-5p含量。Transwell实验检测3组细胞迁移能力和侵袭能力。Western-blot法检测3组细胞神经-钙粘素（N-cadherin）、α-连环蛋白（α-Catenin）和泛素水解酶22（USP22）表达水平。运用生物信息学方法预测miR-30a-5p的靶基因。采用Western blot法检测USP22过表达对miR-30a-5p抑制EMT的拮抗作用。双荧光素酶实验检测miR-30a-5p与USP22的关系。建立皮下移植瘤模型观察miR-30a-5p的体内作用。【结果】30a-5p组宫颈癌细胞miR-30a-5p的表达水平明显上调,表达水平为Control组的853.82（862.26~843.11）倍（P＜0.01）。30a-5p组侵袭细胞数量8.17（8.32~8.03）明显低于Control组（P＜0.01）。30a-5p组细胞N-cadherin蛋白的细胞内含量明显下降,α-Catenin蛋白的细胞内含量明显上升,USP22蛋白表达量明显降低。合并USP22过表达处理的30a-5p组宫颈癌细胞中N-cadherin蛋白表达量明显升高,α-Catenin蛋白表达量明显降低。双荧光素酶检验结果显示USP22为miR-30a-5p的下游靶基因（P＜0.01）。30a-5p组皮下移植瘤明显小于Control组（P＜0.01）。与Control组肿瘤组织相比,30a-5p组肿瘤组织miR-30a-5p的相对含量升高,USP22蛋白含量降低,N-cadherin蛋白的含量降低,α-Catenin蛋白含量升高。【结论】miR-30a-5p在宫颈癌Hela细胞中,可能通过靶向识别下游靶基因USP22,进而抑制其翻译。最终实现对宫颈癌细胞EMT过程的抑制。     

Influenza A Virus Hemagglutinin and Other Pathogen Glycoprotein Interactions with NK Cell Natural Cytotoxicity Receptors NKp46, NKp44, and NKp30

Natural killer (NK) cells are part of the innate immunity repertoire, and function in the recognition and destruction of tumorigenic and pathogen-infected cells. Engagement of NK cell activating receptors can lead to functional activation of NK cells, resulting in lysis of target cells. NK cell activating receptors specific for non-major histocompatibility complex ligands are NKp46, NKp44, NKp30, NKG2D, and CD16 (also known as FcγRIII). The natural cytotoxicity receptors (NCRs), NKp46, NKp44, and NKp30, have been implicated in functional activation of NK cells following influenza virus infection via binding with influenza virus hemagglutinin (HA). In this review we describe NK cell and influenza A virus biology, and the interactions of influenza A virus HA and other pathogen lectins with NK cell natural cytotoxicity receptors (NCRs). We review concepts which intersect viral immunology, traditional virology and glycobiology to provide insights into the interactions between influenza virus HA and the NCRs. Furthermore, we provide expert opinion on future directions that would provide insights into currently unanswered questions.     

Atrial Fibrillation Clinics in Canada: A Nationwide Project Report

The increased prevalence of atrial fibrillation (AF) has led to specialized AF clinics (AFCs) to facilitate management of AF patients. In this article we report on outpatient AFCs in Canada, which is essential to health policies required to standardize the performance of existing AFCs and help design new AFCs. We surveyed 14 clinics in 5 provinces; 100% provided responses to a detailed questionnaire on clinic processes and care practices. Fourteen care maps were analyzed, and 5 models of care were identified; 4 were specific to AFCs. An online survey with 49 questions included items on: (1) process before visit; (2) process at visit; (3) patient education provided; (4) outreach; and (5) specific clinic information. Clinicians’ advice to patients on self-care items such as: (1) amount of alcohol and (2) caffeine intake; (3) exercise activity; (4) stressful events; (5) “when to go to the emergency department”; and (6) lifestyle changes, were evaluated to assess consistency in practice. There were moderate variances in clinicians’ advice to patients in 5 of 6 self-care items. The 1 item that had 100% consistent practice recommendation was when to go to the emergency department. A guideline-based clinical assessment checklist (CAC) was piloted to obtain feedback on its usability in real-world practice; revisions finalized the “simplified CAC” for AF care encompassing 35 data points with rationale. There was 100% positive feedback on its ability to provide baseline elements in AF care. When validated, a “simplified CAC” can facilitate a standardized clinical assessment tool in clinical practice.