腺病毒介导MDA-7/IL-24选择性杀伤肝癌 细胞HepG2的研究

目的 ：观察MDA-7/IL-24基因对肝癌的选择性杀伤作用，为肝癌的基因治疗提供理论基础。 方法 ：将携带人MDA-7/IL-24基因的腺病毒Ad.mda-7感染人正常肝细胞L02和肝癌细胞HepG2;用RT-PCR法观察MDA-7/IL-24基因的表达;ELISA方法检测细胞培养上清液中MDA-7/IL-24蛋白的浓度;4甲基偶氮唑蓝染色法（MTT）及Hoechst染色观察MDA-7/IL-24对肝癌细胞的生长抑制和杀伤作用;Annexin-V和PI双染后流式细胞仪检测2种细胞的凋亡;用流式细胞仪检测细胞周期。 结果 ：复制缺陷型腺病毒能介导外源基因MDA-7/IL-24在肝癌细胞株HepG2和正常细胞L02中的高效表达;细胞培养上清液中有MDA-7/IL-24蛋白的表达; MDA-7/IL-24能明显抑制肝癌细胞生长并可促进肝癌细胞的凋亡;MDA-7/IL-24阻滞肝癌细胞于G2/M期，能选择性杀伤肝癌细胞而对正常的肝细胞无阻滞作用和毒性作用。结论 ：复制缺陷型重组腺病毒载体Ad.mda-7能介导MDA-7/IL-24基因在人肝癌细胞中高效表达，促使细胞增殖阻滞及诱导肿瘤细胞凋亡，选择性地杀伤肝癌细胞HepG2，而对正常肝细胞L02无任何毒性作用。     

三磷酸腺苷结合盒运转体A1与颈动脉粥样硬化斑块的关系

目的 探讨三磷酸腺苷结合盒运转体A1(ATP binding cassette transporter A1,ABCA1)在人颈动脉粥样硬化斑块中的表达变化及作用机制.方法 收集24例人颈动脉粥样硬化斑块标本和10例肠系膜动脉标本(对照组),采用RT-PCR测定ABCA1 mRNA和视黄酸X受体α(RXRα)mRNA表达水平,并采用Western Blot检测ABCA1及RXRα的蛋白表达水平.24例人颈动脉粥样硬化斑块标本按病理分级,比较病理组织为Ⅲ级和Ⅰ级动脉粥样硬化组织间ABCA1 mRNA、RXRαmRNA表达水平及蛋白表达水平.结果 颈动脉粥样硬化斑块组的ABCA1 mRNA(0.79±0.04)和RXRα mRNA(0.73±0.04)表达与对照组相比上调,差异有统计学意义(P＜0.05);ABCA1 mRNA与RXRα mRNA增加水平相关(P＜0.05);颈动脉粥样硬化斑块的ABCA1蛋白表达(0.22±0.03)下调水平与对照组(0.53±0.03)相比差异有统计学意义(P＜0.05);Ⅲ级和Ⅰ级动脉硬化斑块ABCA1mRNA、RXRα mRNA及蛋白表达水平差异有统计学意义(P＜0.05).结论 ABCA1及RXRα蛋白表达水平下调可能是进展性动脉粥样硬化损害的关键因素.     

对血清蛋白白球比值参考范围的修改及其临床应用

目的探讨血清蛋白白球比值(A／G)的参考范围和A／G在计算过程中产生的误差对临床应用的影响。方法①选取健康查体人员统计其A／G参考范围；②从理论上分析A／G计算误差的存在。结果①以统计结果的2s置信区间作为A／G的参考范围是1.1～2.2；②A／G计算值的误差会远大于相应总蛋白和白蛋白的测定误差。结论A／G较合适的参考范围为1.1～2.2,并且在临床应用中仍可根据实际情况降低其范围的下限。     

Safety and efficacy of botulinum toxin type A following long-term use

Botulinum toxin serotype A (BoNT-A) has long heritage of use leading to confidence in its safety and efficacy. The application of BoNT-A does not lead to persistent histological changes in the nerve terminal or the target muscle. Clinical trials defined the safety and tolerability profile of BoNT-A across common therapeutic indications and showed an incidence of adverse events of approximately 25% in the BoNT-A-treated group compared with 15% in the control group. Focal weakness was the only adverse event to occur more often following BoNT-A treatment. Long-term BoNT-A administration has been assessed in various treatment settings, with the level and duration of BoNT-A efficacy response being maintained over repeated rounds of injection with no major safety concerns. The treatment of children with cerebral palsy often require long-term, repeated, multimuscle BoNT-A injections that lead to the administration of comparably higher toxin doses. Despite the high total body doses used, their distribution over multiple muscles and injection sites means that systemic side effects are rare. Recent formulation changes have reduced the incidence of antibody development following treatment with BOTOX^®. These findings show long-term BoNT-A treatment to be both safe and efficacious for a wide variety of indications.     

Matrix metalloproteinases-2 and -9 in cervical cancer: different roles in tumor progression

The incidence of uterine cervical cancer has increased slightly in Western countries, with an increase in relatively young women. Overexpression of matrix metalloproteinases (MMPs)-2 and -9 has turned out as a prognostic factor in many cancers. We compared the expression of the proteins MMP-2 and MMP-9 in cervical primary tumors with clinical outcome and risk factors of cervical cancer. One hundred sixty-one patients with cervical cancer treated in Ume? University Hospital or Sahlgrenska University Hospital, Sweden, between 1991 and 1995 were included in the study. Paraffin-embedded tissue samples obtained prior to treatment were examined immunohistochemically by specific antibodies for MMP-2 and MMP-9. Forty-two percent of the tumors were intensively positive for MMP-2 and 31% for MMP-9. Nineteen percent of the samples were intensively positive for both proteinases and 47% negative or weak for both. Overexpression of MMP-2 seemed to predict unfavorable survival under Kaplan-Meier analysis and in the multivariate analysis. Early sexual activity and low parity seemed to correlate to overexpression of MMP-2. MMP-9 was not associated with survival or sexual behavior. Intensive MMP-9 was noted in grade 1 tumors. We conclude that MMP-2 and MMP-9 have different roles in uterine cervical cancer. MMP-2 could be associated with aggressive behavior, but MMP-9 expression diminishes in high-grade tumors.     

Digestive properties of the venom of the Australian Coastal Taipan, Oxyuranus scutellatus (Peters, 1867).

The digestive properties of Australian elapid snake venoms have not been studied to any great extent. To address this, the in vitro digestive properties of Oxyuranus scutellatus (Australian Coastal Taipan) venom were investigated in a simulation of the in vivo conditions using the parameters reported for the stomach of snakes and representative prey for this species. The amount of soluble protein released was measured over time using a bicinchoninic acid (BCA) assay. Dismembered mouse hindlegs were injected intramuscularly with 0.1 ml O. scutellatus venom (concentration 10 mg/ml) and maintained in a micro-anaerobic, acidic environment (pH approximately 1.2-1.7) at 25 degrees C. The bathing liquid was sampled every 24 h for 7 days, and assayed for soluble protein. Statistical analysis revealed that O. scutellatus venom increased the rate at which proteins were released when compared to a negative control suggesting the potential importance of envenomation in the digestion of whole prey.     

鸡抗内毒素卵黄抗体IgY的制备

目的：研究分别应用内毒素（LPS）、类脂A(LipidA)和大肠杆菌突变株15免疫鸡后其蛋黄中抗内毒素抗体IgY的产量、纯度、效价并筛选最佳免疫抗原。方法：分别应用内毒素（LPS）、类脂A(LipidA)和大肠杆菌J5突变株作为抗原免疫25周龄Leghom鸡，水溶法（WD）提取蛋黄中抗体IgY，双紫外光测定抗体含量，SDS-PSGE电泳检测抗体纯度，细胞酶联染色和ELISA检测抗体特异性、效价及筛选最佳免疫抗原。结果：3种抗内毒素IgY含量和效价分别为14.4mg/ml和1：12 800（J5）、10.61mg/ml和1：12 800（LPS）、9.26mg/ml和1：3200（LipidA），抗体纯度均为95％左右。结论：大肠杆菌突变株J5和内毒素（LPS）为最佳免疫抗原，免疫鸡后其蛋黄中抗内毒素抗体IgY的产量和效价最高。     

从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位

目的：建立一种有效从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位HspA的方法。方法：重组基因工程大肠杆菌发酵后，表达的Hp r HspA包涵体经洗涤，变性，复性，采用Q Sepharose High Performance阴离子交换层析和Superdex75凝胶过滤层析分离纯化，使用SDS－PAGE和HPLC检测纯度，选用ELISA和动物实验对纯化蛋白的免疫学活性和生物学活性进行鉴定。结果：Hp的HspA包涵经洗涤和溶解后，Hp的HspA的纯度＞60％，包涵体溶解液经阴离子交换层析和凝胶过滤层析后纯度超过95％，Hp的HspA纯品经检测具有良好的免疫学活性和生物学活性。结论：本研究建立的分离纯化方法可从包涵体中获得高纯度的重组HspA蛋白，为进一步的动物实验研究奠定了基础。     

Haemophilia A and haemophilia B: molecular insights

This review focuses on selected areas that should interest both the scientist and the clinician alike: polymorphisms within the factor VIII and factor IX genes, their linkage, and their ethnic variation; a general assessment of mutations within both genes and a detailed inspection of the molecular pathology of certain mutations to illustrate the diverse cause–effect relations that exist; a summary of current knowledge on molecular aspects of inhibitor production; and an introduction to the new areas of factor VIII and factor IX catabolism. An appendix defining various terms encountered in the molecular genetics of the haemophilias is included, together with an appendix providing accession numbers and locus identification links for accessing gene and sequence information in the international nucleic acid databases.