新药研制中常见问题浅析

自1985年我国实施《新药审批办法》以来,我国的新药研究取得了很大的成绩,为防病治病作出了重大贡献。我省新药研制在全国处于领先地位,每年批准的新药占全国批准新药总数的10％以上。十多年来新药研制的技术要求已逐步规范化,但由于各研制者对新药研究技术要求掌握程度的差别,及申报经验等方面的原因,在新药研制过程中出现各种问题,造成报批的失败或延误。本文通过对本单位已批准的40份新药证书和10多个新药在研制过程中出现的常易发生的问题作一简要的分析,以供新药研制者参考。 对本单位已取得的新药证书进行统计表明,从研制立项开始到获得新药证书,二类西药、三类西药和四类西药的平均研制时间分别为5.5(3.5～7)、     

经脐单孔与常规腹腔镜阑尾切除手术的比较研究

目的:探讨单孔腹腔镜阑尾切除手术的可行性。方法:对2011年1月—2011年12月收治的122例急性阑尾炎病例,按照随机原则分单孔腹腔镜组(61例)和常规腹腔镜组(61例),对两组的手术时间、术中出血量、肠道功能恢复时间、住院时间、并发症、中转率进行比较。结果:两组在术中出血量、术后肠道功能恢复时间、住院时间及术后并发症的比较,差异均无统计学意义(P>0.05),单孔腹腔镜组因粘连(2例)和腹膜后阑尾(1例)中转常规腹腔镜手术;坏疽性(穿孔性)阑尾炎的手术时间,单孔腹腔镜组明显长于常规腹腔镜组(P=0.004);单纯性阑尾炎和化脓性阑尾炎两组的手术时间无统计学差异(P>0.05)。结论:选择性对急性阑尾炎行单孔腹腔镜手术治疗是安全可行的;与常规腹腔镜比较,单孔腹腔镜治疗单纯性阑尾炎和化脓性阑尾炎同样具有恢复快、创伤小的优点,且比常规腹腔镜有更好的维护形体的效果。     

氯沙坦联合表没食子儿茶素没食子酸酯对大鼠糖尿病肾病治疗的研究

目的观察氯沙坦联合表没食子儿茶素没食子酸脂（EGCG）对糖尿病肾病大鼠的肾脏保护作用。方法采用链脲佐菌素（60mg·kg^-1）腹腔注射建立糖尿病大鼠模型,模型成功后分为正常组、糖尿病组、氯沙坦组、EGCG组、氯沙坦＋EGCG组。EGCG组和氯沙坦＋EGCG组每周给予ECd2G10mg·kg^-1腹腔注射,氯沙坦组和氯沙坦＋EGCG组每天给予氯沙坦40rag·kg^-1灌胃。分别测定各组4、8、12周时血肌酐（Cr）、尿素氮（BUN）、葡萄糖（GLU）、甘油三酯（TG）、总胆固醇（CHOL）、高低密度脂蛋白（HDL）、低密度脂蛋白（LDL）、白蛋白（Alb）、24h尿蛋白定量（24hPro）。应用RealtimePCR法检测肾皮质活性氧（ReactiveOxygenSpecies,ROS）产生的主要通路NADPH氧化酶Nox4mRNA表达、Westernblot法检测肾皮质蛋白NADPH氧化酶Nox4的表达。结果糖尿病组相对于正常对照组Cr,BUN,TG,CHOL,HDL,LDL,24hPro水平均显著升高（P〈0．05）。与糖尿病组比较,氯沙坦组、EGCG组、氯沙坦＋ECa2G组大鼠Cr、24hPro有显著减少（P〈0．05）,但GLU,BUN,TG,CHOL,HDL,LDL均无明显减少（P〉0．05）。与氯沙坦组和EGCG组比较,氯沙坦＋ECA2G组Cr,GLU,BUN,TG,CHOL,HDL,LDL,ALB,24hPro均有差异但差异不明显（P〉0．05）。在第4和12周与正常组比较,糖尿病组大鼠肾小球硬化明显;与糖尿病组比较,氯沙坦组、EGCG组、氯沙坦＋EGCG组大鼠肾小球硬化均显著改善;与氯沙坦组比较,EGCG组、氯沙坦＋EGCG组大鼠。肾小球硬化缓解更加明显;在电子显微镜观察下与正常组比较,糖尿病组大鼠肾小球足细胞明显融合,氯沙坦组少量融合,而EGCG组和氯沙坦＋EGCG组足细胞基本完好。糖尿病组Nox4蛋白表达最高;与EGCG组和氯沙坦组比较,氯沙坦＋EGCG组Nox4蛋白表达均降低,并且随着时间的推移降低更加显著（P〈0．05）。第4、8、12周NADPH氧化酶Nox4mRNA表达均降低,但不同时间点同组之间差异不是很明显（P〉0．05）。结论EGCG协同氯沙坦通过下调NAD—PH氧化酶Nox4的表达,抑制氧化应激对糖尿病大鼠肾脏的损伤,对糖尿病肾病具有良好的保护作用。     

医院客户服务绩效评价与提升策略研究

本文通过对客户服务的绩效管理流程进行论述,并对绩效管理体系改革措施的效果进行评价,以建立学习型组织和组织的绩效文化,形成具有激励作用的工作氛围为目的。本院参考国内外经验,结合自身实际,对医院员工的客户服务绩效进行考核,提升医院创新能力和管理水平,为医院在市场竞争环境下获取竞争优势提供思路。     

内注外剥治疗内外痔临床观察

痔疮是一种常见病。目前国内外治疗内痔、外痔方法很多。我院曾用北京中医研究院研制的“消痔灵”注射液治疗内痔同手术剥离或切除外痔的综合治疗收到很好的治疗效果。1 临床资料我院在1992—1994年治疗内痔与外痔并存患者100例。其中男68例。女32例,年龄最小19岁,26岁以上者占80％,病史最长35年,最短者1年,平均病史6年。临床特征:肛     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

ARBs和活性维生素D在糖尿病肾病治疗中的新进展

糖尿病肾病作为糖尿病的微血管并发症,是导致终末期肾病的主要原因之一.肾素血管紧张素(RAS)在糖尿病肾病肾脏损伤过程中扮演重要作用.RAS抑制剂在临床广泛使用,但却会引起肾素补偿性增加,从而限制了RAS抑制剂的疗效.维生素D通过抑制肾素表达从而能负性调节RAS.RAS抑制剂联合维生素D类似物能显著改善糖尿病肾病的肾脏损...     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.