白蛋白治疗对小鼠脑缺血早期脑组织Toll样受体4和骨髓分化因子88 mRNA表达的影响

目的研究人血白蛋白治疗对局灶性脑缺血再灌注小鼠脑组织Toll样受体4(TLR4)和骨髓分化因子88(MyD88)mRNA表达的影响。方法健康雄性成年昆明小鼠54只,随机分为假手术组(14只)、生理盐水组(20只)和白蛋白组(20只),每组又均分为再灌后6、24 h 2个时间点。采用左侧大脑中动脉线栓法制备小鼠局灶性脑缺血再灌注模型,RT-PCR法检测左侧脑组织TLR4、MyD88 mRNA的表达水平,并进行神经功能缺损评分。结果生理盐水组和白蛋白组脑缺血再灌注后6、24 h TLR4、MyD88 mRNA表达水平明显高于假手术组,且24 h表达水平升高更明显(P<0.05);但白蛋白组6、24 h TLR4、MyD88 mRNA表达量明显低于生理盐水组(P<0.05);24 h白蛋白组小鼠的神经功能缺损评分与生理盐水组比较有明显改善(P<0.05);24 h TLR4、MyD88 mRNA表达水平与神经功能缺损评分呈正相关(r=0.92,r=0.85,P<0.05)。结论白蛋白治疗可以下调脑缺血早期脑组织高表达的TLR4、MyD88 mRNA的水平,改善脑缺血后小鼠神经功能缺损。     

乳腺癌组织中PSA mRNA和VEGF mRNA的表达及其关系

目的探讨前列腺特异抗原(PSA)mRNA和血管内皮细胞生长因子(VEGF)mRNA在乳腺癌组织中的表达及其关系。方法采用半定量逆转录聚合酶链式反应(RT-PCR)检测35例乳腺癌、12例癌旁乳腺组织和10例乳腺纤维腺瘤组织中的表达,免疫组化检测35例乳腺癌组织中ER、PR的表达,分析PSA mRNA和VEGFmRNA表达之间的关系。结果PSA mRNA在乳腺癌组织中的表达水平明显低于癌旁乳腺组织和乳腺纤维腺瘤组织,差异有显著性(P值均=0.00)。VEGF 121、165mRNA在乳腺癌中的表达水平明显高于癌旁乳腺组织和乳腺纤维腺瘤组织的表达(P值均=0.00)。ER、PR阳性表达的乳腺癌组织中PSA mRNA表达高于ER、PR阴性者,差异有显著性(P=0.001和0.004)。PSA mRNA表达阳性者VEGF 121、165mRNA表达水平明显低于PSA mRNA表达阴性者(P=0.034、0.026)。结论PSA mRNA在乳腺癌组织中的表达下调,但其表达受ER、PR的调控,而且其可能具有抑制乳腺肿瘤血管生成的作用。     

醛固酮对人近端肾小管上皮细胞结缔组织生长因子mRNA表达的影响

目的探讨醛固酮(ALD)在体外是否能够促进人近端肾小管上皮细胞株(HK2)结缔组织生长因子(CTGF)的表达.方法采用体外培养的HK2细胞,分别观察不同浓度(10-11、10-10、10-9、10-8、10-7 mol/L)ALD处理48 h,或用10-9 mol/L ALD处理不同时间(24 h、48 h、72 h)对HK2细胞CTGF表达的影响,应用RT-PCR法检测CTGFmRNA水平的表达.结果HK2细胞培养48 h后,空白对照组即有CTGF的基础表达,ALD不同浓度组CTGF mRNA的表达均有增加,与空白对照组相比较差异有统计学意义(P＜0.05或P＜0.01);以10-9 mol/L浓度的ALD作用于HK2细胞,随着作用时间的延长,CTGF mRNA的表达量明显增加,与对照组相比较差异有统计学意义(P＜0.05或P＜0.01).结论ALD能够促进HK2细胞CTGF mRNA水平的表达,并具有剂量依赖性和时间依赖性,ALD可能通过促进CTGF的分泌,发挥其促进肾间质纤维化的作用.     

Edible safety requirements and assessment standards for agricultural genetically modified organisms.

This paper describes the background, principles, concepts and methods of framing the technical regulation for edible safety requirement and assessment of agricultural genetically modified organisms (agri-GMOs) for Shenzhen Special Economic Zone in the People's Republic of China. It provides a set of systematic criteria for edible safety requirements and the assessment process for agri-GMOs. First, focusing on the degree of risk and impact of different agri-GMOs, we developed hazard grades for toxicity, allergenicity, anti-nutrition effects, and unintended effects and standards for the impact type of genetic manipulation. Second, for assessing edible safety, we developed indexes and standards for different hazard grades of recipient organisms, for the influence of types of genetic manipulation and hazard grades of agri-GMOs. To evaluate the applicability of these criteria and their congruency with other safety assessment systems for GMOs applied by related organizations all over the world, we selected some agri-GMOs (soybean, maize, potato, capsicum and yeast) as cases to put through our new assessment system, and compared our results with the previous assessments. It turned out that the result of each of the cases was congruent with the original assessment.     

标记TRAILR地高辛探针及原位杂交检测喉癌组织mRNA

目的:标记肿瘤坏死因子相关凋亡诱导配体受体(TRAILR)地高辛(DIG)探针,检测喉癌组织中TRAILR mRNA,并探讨其意义.方法:采用RT-PCR原位杂交法检测喉癌组织TRAILR mRNA. 结果:TRAILR地高辛探针标记成功;喉癌和正常对照中死亡受体4、5(DR4,DR5)mRNA呈强阳性,阳性率均为100%(34/34,5/5);正常对照中诱捕受体2(DcR2)mRNA呈强阳性,阳性率100%(5/5);喉癌中DcR2 mRNA可检出,阳性率14.71%(5/34);DcR2 mRNA主要分布在癌巢组织边缘,强度由癌巢中央到癌巢边缘逐渐加强.结论:TRAIL受体基因可能在喉鳞状细胞癌凋亡调控机制中发挥重要作用.     

Expression of hepatic thrombopoietin mRNA in primary cultured hepatocytes and in rats with acute liver injury or bone marrow suppression with or without cirrhosis

BACKGROUND AND AIMS: The main causes of thrombocytopenia in cirrhosis are thought to be platelet destruction and the reduction of thrombopoietin (TPO) expression in the liver. The mechanisms by which levels of TPO mRNA are regulated in cirrhosis have not been elucidated. In this study, we investigated some possible mechanisms. METHODS: We used three experimental models: bone marrow suppression, acute liver injury and primary cultured hepatocytes. We used northern blots to assess the kinetics of TPO mRNA expression in the livers of irradiated rats (with and without cirrhosis) in acute liver injury and in primary cultured hepatocytes treated with hepatotoxin or cytokines. RESULTS: Although the bone marrow was hypocellular, there was no apparent enhancement of TPO mRNA expression in the irradiated rats with cirrhotic livers compared with the unirradiated rats with cirrhotic livers. There were no conspicuous changes in hepatic TPO mRNA expression between the livers of the control rats and the three models of acute liver injury. There were no conspicuous changes in the levels of TPO mRNA between control hepatocytes and hepatocytes treated with hepatotoxin or cytokines. CONCLUSIONS: Our results suggest that bone marrow is not a regulator of hepatic TPO production in cirrhosis. The reduced TPO mRNA expression found in cirrhotic rats may not result merely from serious cellular damage; it may be associated with cirrhosis-specific regulatory mechanisms for the expression of the TPO gene. Further studies are needed to search for other factors that may induce reduced TPO expression.     

成人睾丸中一种新的calpastatin同源蛋白的克隆和分析

目的:克隆人精子发生相关的基因.方法:使用人胚胎和成人睾丸cDNA探针与本室制作的人睾丸dDNA微阵列杂交,差异杂交的克隆进行序列测定和分析.结果:发现Calpastatin在睾丸中表达的一种新的同源蛋白,在睾丸中高度表达.结论:calpastatin在睾丸中是以一种新的同源蛋白形式表达,可能与精子发生相关.     

离子辐射通过caspase途径而非p53途径诱导Jurkat细胞凋亡

目的:研究离子辐射诱导Jurkat细胞凋亡的动态趋势及其机制。方法:分别以5、10、20Gy的辐射量照射Jurkat细胞,其中一份样本在照射前加入zVAD.fmk,在照射后培养6、12、24、36和48h收获细胞,应用AnVPI双染和呈现subG1峰技术,在流式细胞仪上定量测定凋亡细胞。采用WesternBlot技术检测p53蛋白的表达。结果:细胞受照射后,凋亡细胞数随培养时间的延长和辐射量的加大而增加。在6h,只有20Gy才诱导细胞显著凋亡(P<0.005),5Gy的照射要到24h才出现明显的凋亡(P<0.05)。在48h,20Gy照射诱导的凋亡细胞数是所有时间点和剂量强度中最高的(P<0.001)。加有zVAD.fmk的细胞,尽管都是10Gy照射,比未加zVAD.fmk的细胞表现出凋亡细胞明显减少(P<0.005),但与未加zVAD.fmk、照射量是5Gy的细胞相比,凋亡细胞数仍显著增加(P<0.005)。Jurkat细胞在照射前后都没有p53表达。结论:离子辐射诱导细胞凋亡呈现出时间和剂量效应关系;capase抑制剂zVAD.fmk部分抑制离子辐射诱导细胞凋亡;离子辐射诱导细胞凋亡的机制独立于肿瘤抑制基因p53,caspase可能在其中起重要作用。