肉碱和长链脂肪醇对小鼠大脑乙酰胆碱的影响

目的　探讨肉碱和长链脂肪醇对小鼠大脑乙酰胆碱 (Ach)含量的影响。方法　用析因实验设计方法将BALB/C成年雄性小鼠随机分为标准对照、高脂对照、肉碱、长链脂肪醇、肉碱和长链脂肪醇合用共 5组 ,标准对照组饲标准饲料 ,其它各组饲含 2 0 %油脂的饲料 ,肉碱 [剂量 50mg/ (kg·鼠重 ) ]和长链脂肪醇 [剂量 50mg/ (kg·鼠重 ) ]以0 0 9%的剂量加到饲料中连续喂养 3 0d后 ,称体重、脑重 ,计算脑体比 ,用碱性羟胺比色法测大脑组织中Ach含量。结果　肉碱和长链脂肪醇对成鼠体重、脑重、脑体比无显著影响 (P >0 0 5) ;高脂饲料组小鼠脑内Ach含量较对照组明显降低 (P <0 0 5) ,长链脂肪醇可引起高脂动物脑内Ach含量升高 (P <0 0 5) ,但与正常饲料组无明显差异 (P >0 0 5) ;肉碱引起脑内Ach含量明显增多 (P <0 0 5) ;但两者联合作用时有交互作用 (P <0 0 1) ,且脑内Ach含量约等于者单独作用的差值。结论　肉碱、长链脂肪醇能分别促进小鼠大脑Ach含量的增加 ,但联合作用时表现为拮抗     

反相高效液相色谱法测定托拉噻米胶囊中托拉噻米的含量

目的建立高效液相色谱法测定托拉噻米胶囊中托拉噻米的含量。方法以岛津Sh impack CLC(M)C18(250 mm×4.6 mm,5μm)为色谱柱,以甲醇-0.02 mol.L-1磷酸二氢钾溶液(用磷酸调节pH至3.0)(65∶35)为流动相,检测波长为UV291 nm,流速为0.9 ml.min-1。结果托拉噻米在2~24μg.ml-1范围内具有良好线性关系(r=0.999 9),平均加样回收率为99.73%,相对标准差为0.17%;三批样品的标示百分含量(%)分别为99.8、100.1及100.4。结论本方法准确、灵敏度高、重现性好,可用于托拉噻米胶囊的含量测定。     

Tissue microarray: an effective high-throughput method to study the placenta for clinical and research purposes.

OBJECTIVE: Tissue microarray (TMA) technology allows simultaneous examination of the expression of many molecular markers (protein, mRNA, DNA, etc.) with high-throughput. The application of this technology, to date, has been largely confined to the study of cancer. Placental pathology poses unique challenges because of the size of the organ, its complex anatomy, as well as its histological heterogeneity. The objective of this study was to assess the feasibility and efficiency of TMAs for immunohistochemistry and in situ hybridization of placental tissues. STUDY DESIGN: TMAs were constructed using an automated tissue arrayer. Standard 0.6-mm or 1-mm microarray needles were used. Villous parenchyma, basal plate, and chorioamniotic membranes were targeted in each block. Five mum-thick TMA sections underwent immunohistochemical analysis of both cytoplasmic and nuclear antigens using a panel of antibodies against a variety of cytoplasmic [cytokeratin-7, vascular endothelial growth factor (VEGF), and protein Z], membranous (endoglin), and nuclear (c-fos and c-jun) antigens. mRNA in situ hybridization for surfactant protein A (SP-A) and chromogenic in situ hybridization for the Y chromosome (DYZ1) were also performed. RESULTS: Validation of TMA immunoreactivity demonstrated comparable results with corresponding whole sections. When a two-tiered scoring system (positive/negative) was employed, there was agreement between two and three cores and whole tissue sections (kappa>0.7). When a three-tiered scoring system (negative, weak-positive, or strong-positive) was used, the data from three cores showed the highest agreement with whole tissue sections (kappa >0.7). In situ hybridization experiments for mRNA and DNA were also successful in that the signals were readily detectable. Successful transfer from the donor block to the recipient block differed according to the anatomical compartment. The transfer efficiency of villous parenchyma, basal plate, and chorioamniotic membranes were 96.9% (875/903), 76.7% (115/150), and 75.4% (224/297), respectively. CONCLUSION: TMA is a practical and effective tool for high-throughput molecular analysis of the human placenta. Duplicate and triplicate cores offer agreement with whole tissue sections for two-category distinction immunostaining. TMA also affords relevant results from in situ hybridization experiments for mRNA and DNA. The major advantages are the conservation of tissues and reagents, simultaneous comparison of molecular markers in different anatomical compartments of the placenta, and reduction of experimental error.     

人表皮生长因子对牛角膜内皮细胞增殖和细胞周期的影响

目的 观察表皮生长因子(EGF)对牛角膜内皮细胞增殖和细胞周期的影响.方法 采用体外培养的牛角膜内皮细胞,在培养液中分别添加不同浓度的人表皮生长因子(hEGF),加药后第3、7天采用MTT法,在酶标仪492 nm波长处测定吸光度值观测细胞增殖,并记录细胞形态变化.将体外培养的牛角膜内皮细胞分为二组,常规DMEM培养液组和含50 ng/mL EGF的DMEM培养液组,培养3、7天后,应用流式细胞仪检测细胞周期各阶段G1、S、G2期分布.结果 与对照组相比,不同浓度EGF组对牛角膜内皮细胞增殖有促进作用且与浓度相关,其中10 ng/mL和100 ng/mL组作用强于1 ng/mL组.EGF加入后3 d,对照组S期细胞24.5%,G2-M期细胞0.08%,加药组S期细胞24.6%,G2-M期细胞0.06%.与对照组相比较,各期细胞分布大致相等,无显著差异.加入7 d后对照组S期细胞20.8%,G2-M期细胞0.41%,加药组S期细胞18.2%,G2-M期细胞1.55%,细胞分布发生明显变化.结论 EGF促进体外培养的牛角膜内皮细胞增殖,并使体外培养的牛角膜内皮细胞的细胞周期发生变化,S期细胞比例下降.     

Attempted suicide and polymorphism of the serotonin transporter gene in Chinese patients with schizophrenia

Abnormalities of serotonin synthesis and metabolism may be associated with suicidality. The serotonin transporter gene (5-HTT) is one of the important genes involved in the regulation of serotonin neurotransmission. We examined the association of suicidal behavior in Chinese schizophrenic patients with a functional polymorphism of the promoter region of the 5-HTT gene (5-HTTLPR). The 5-HTTLPR genotype was determined by polymerase chain reaction for 76 suicidal and 262 non-suicidal patients with a diagnosis of schizophrenia (DSM-IV criteria). All subjects were unrelated to each other, and all were Chinese. There was no significant genotypic or allelic association of the 5-HTTLPR polymorphism with history of attempted suicide. From our results, this 5-HTTLPR polymorphism is unlikely to have a major effect on suicidal behavior in Chinese patients with schizophrenia.     

光茎大黄化学成分研究

目的:对光茎大黄根部的化学成分进行研究.方法:采用硅胶柱层析、重结晶、MS、1HNMR、13CNMR等技术进行分离、结构鉴定.结果:从该植物中分离并鉴定了七个化合物,分别是:正二十六烷酸(n-hexacosnic acid,Ⅰ),棕榈酸(palmitic acid,Ⅱ),胡萝卜甙(daucosterol,Ⅲ),大黄酚甲醚(chrysophanol-8-Me ether,Ⅳ),ω羟基大黄素(citreorosein,Ⅴ),大黄酚-8-O-葡萄糖甙(chrysophanol-8-O-β-D-glucopyranoside,Ⅵ)和2,5-二甲基-7-甲氧基色原酮(2,5-dimethyl-7-methoxychromone Ⅶ).结论:该七个化合物均为首次从该植物中分离得到.     

苍耳子对小鼠脾淋巴细胞蛋白质组影响的双向电泳分析

目的观察苍耳子对致敏小鼠脾淋巴细胞蛋白质表达的影响.方法NIH小鼠正常组腹腔注射生理盐水,造模组和治疗组均注射羊红细胞(SRBC)致敏,治疗组经致敏后以苍耳子水煎剂灌胃,处理4 d后,分离小鼠脾淋巴细胞;提取总蛋白,双向凝胶电泳(2-DE)后,以计算机图像分析软件分析电泳图谱.结果致敏会引起多种蛋白质表达发生变化,而苍耳子可逆转其中部分蛋白质的变化.结论苍耳子可改善由致敏引起的小鼠脾淋巴细胞中部分蛋白表达的变化.     

纯系LEW鼠及裸小鼠膀胱灌注给药方式的探讨

 目的 探讨纯系LEW大鼠及裸小鼠膀胱灌注给药方法。方法 显微镜下解剖雌性LEW大鼠及裸小鼠尿道并根据解剖特征改造套管针。通过插入尿道的套管针膀胱灌注墨水，同时观察插管及膀胱充盈和染色情况。持续观察小鼠和大鼠生存状况及尿道损伤情况。结果 显微镜下可见雌性LEW大鼠及裸小鼠尿道与人存在明显差异。经尿道插管成功率高(100％)，动物生活状况没有明显受到插管灌注影响。结论 裸鼠膀胱灌注给药治疗是一种成功率高而可行的给药手段，动物膀胱灌注化疗模型可能为研究膀胱肿瘤治疗提供更为有效的工具。     

海口地区超广谱β-内酰胺酶阳性菌检测及其流行病学分析

目的为了解海口地区超广谱β-内酰胺酶(ESBLs)阳性菌的种类及其分布,为控制其扩散、预防暴发流行提供科学依据. 方法采用双纸片琼脂扩散法,测定3年中从临床各类标本分离的革兰阴性菌的ESBLs,比较菌种、病房及标本间ESBLs阳性率差异. 结果产ESBLs的菌种已达28种,阳性率最高达60%,常见菌的阳性率均达约50%,且3年间的变化多数无统计学意义;ESBLs阳性菌分离率在病室间的差异不大,门诊患者相对住院患者较低,在标本间的分布存在一定差异. 结论 ESBLs阳性菌已在海口地区广泛存在,并已成为医院及社区感染的主要病原菌,控制其进一步发展已成为一项迫在眉睫的工作.     

不同剂量卡非佐米对多发性骨髓瘤大鼠SDF-1 CXCR4表达的影响及作用机制

目的 探讨不同剂量卡非佐米对多发性骨髓瘤大鼠基质细胞衍生因子 1(SDF 1)和趋化因子受体（CXCR4）表达水平的影响及相关作用机制。方法 随机选取50只健康SD大鼠中10只为空白组,其余40只大鼠进行多发性骨髓瘤模型建造,空白组大鼠不做处理。建模成功后,随机选取10只大鼠作为模型组,其余30只大鼠为给药组,对给药组大鼠采用不同剂量的卡非佐米进行干预,并根据使用剂量将给药组大鼠分为低剂量组、中剂量组、高剂量组,每组各10只,并对低、中、高剂量组大鼠分别采用15、25、35mg/mL的卡非佐米进行干预,21d后处死大鼠,并对其皮下瘤组织进行切片染色观察,测量5组大鼠荷瘤重量平均值、腹水量平均值和肿瘤体积,采用免疫投射比浊法对5组大鼠SDF-1、CXCR4水平进行检测,酶联免疫吸附法检测核因子 κB（NF κB）、白介素-6（IL-6）、肿瘤坏死因子 α（TNF α）和类胰岛素一号增长因子（IGF-1）表达水平,并阐述卡非佐米的治疗机制。结果 中剂量组大鼠的荷瘤重量平均值、腹水量平均值和肿瘤体积明显低于低剂量组和高剂量组,且SDF 1、CXCR4表达水平亦显著低于低剂量组和高剂量组（均P＜0.05）;中剂量组大鼠NF κB、IL-6、TNF α和IGF-1表达水平较低、高剂量组显著降低（均P＜0.05）。结论 卡非佐米能降低SDF-1、CXCR4水平的表达,抑制炎症通路活性来控制瘤细胞增殖,且中剂量卡非佐米的抑制效果更明显,可为临床用药提供一定的理论依据。