Comparison of the unlabeled and labeled pre-mRNA splicing assays in vitro

Pre-mRNA splicing is a fundamental process required for the expression of most metazoan genes. It is carried out by the spliceosome that catalyzes the removal of non-coding intron sequences to ligate exons into mature mRNA prior to transport and translation. The purpose of our study is to explore whether the in vitro unlabeled pre-mRNA splicing assay could be performed as an alternative method of splicing reaction other than the radiolabeled one. Two different splicing methods in vitro , P labeled and unlabeled pre-mRNA as the substrates in the reaction, were investigated. The radiolabeled products were visualized by autoradiography while the unlabeled products were observed by Ethidium Bromide (EB) staining. As a result, although there are more unspecific bands in the EB staining assay than 32P labeled one, the RNA products of in vitro splicing could be observed clearly. This suggests that the unlabeled pre-mRNA splicing assay can be an optional substitution for the isotope-labeled assay.     

Application of antigens fromTaenia hydatigena cyst fluid for the immunodiagnosis of human hydatidosis

Fluid was collected from cysts ofTaenia hydatigena in 60 adult sheep and fluid from each animal pooled separately. By double diffusion antigen 5 was demonstrated in all pools but one. The criteria are described for selection and standardization of these preparations for use as antigens for the immunodiagnosis of human hydatid disease. Sera from 50 persons harbouring hydatid cysts and from 50 patients with other disease conditions were examined by the arc-5 double-diffusion test, using two antigens prepared fromEchinococcus granulosus andT. hydatigena cyst fluids, respectively. The results showed that a higher diagnostic sensitivity was obtained with the hydatid antigen. The significance of the findings is discussed in terms of their application to human immunodiagnosis in areas where hydatidosis, but not cysticercosis, is rare in livestock.     

国外乳腺癌病理诊断报告标准化概述及对国内标准化报告的建议

随着乳腺癌治疗方案的不断改进和发展,临床医师对乳腺癌病理诊断报告的内容和标准有了更多和更高的要求.     

IFN-γ在沙眼衣原体呼吸道感染中免疫防御机制的探讨

目的:检测与IFN-γ作用相关的酶吲哚胺2,3二氧化酶(IDO)、诱导性一氧化氮合成酶(iNOS)和NADPH氧化酶(ox)gp91在沙眼衣原体呼吸道感染中的表达及与机体防御的关系,探讨衣原体感染中IFN-γ免疫防御作用的机制.方法:用沙眼衣原体小鼠肺炎株(MoPn)通过鼻腔感染C57BL/6(H-2b)小鼠,用过氧化物酶连接的鼠抗衣原体脂多糖单抗染色HeLa 229细胞,检测衣原体在肺组织的生长;用RT-PCR检测衣原体感染后第7及14天小鼠肺组织IFN-γ、IDO、iNOS和gp91NADPH ox mRNA表达.结果:MoPn呼吸道感染后小鼠肺组织匀浆衣原体活性测定,于感染后第2天,HeLa 229细胞内可见有衣原体包涵体生长,IFU值增高,于感染后第7天IFU达最高水平,以后逐渐下降,至感染后21天基本恢复到基线水平.与未感染的对照组比较,Th1细胞因子IFN-γ于感染后第7天表达显著增高,感染后14天有所降低,但仍维持较高水平;同时衣原体感染可显著诱导与IFN-γ作用相关的三种酶IDO、iNOS和gp91 NADPH ox在小鼠肺组织的表达,感染后第7及14天,IDO,iNOS及gp91 NADPH ox的表达与对照组比较均有显著差异,其中IDO和gp91 NADPH ox于感染后第7天mRNA表达增高显著(P＜0.01),14天略有下降(P＜0.05).结论:衣原体呼吸道感染诱导Th1细胞因子IFN-γ mRNA高表达,参与宿主对衣原体的清除及机体免疫防御,此作用可能与其相应的酶IDO、iNOS和gp91 NADPH ox表达增高有关.     

人λ-干扰素在BHK-21中的表达及生物学活性的研究

目的:研究HuIFN-λ1和HuIFN-λ2真核细胞重组表达及其生物学活性.方法:用水疱性口炎病毒(VSV)刺激人宫颈癌HeLa细胞, 用RT-PCR克隆HuIFN-λ1和HuIFN-λ2基因, 与PCAGG-EGFP真核表达载体连接, 构建重组体PCAGG-HuIFN-λ1和PCAGG-HuIFN-λ2并进行鉴定, 后在BHK-21细胞进行表达, 采用VSV*GFP于人肺腺癌A549上进行表达产物抗病毒活性测定;并通过已构建的MDBK-Mxp-Luc细胞系对HuIFN-λ1和HuIFN-λ2诱导MxA抗病毒蛋白产生的特性进行研究.结果:HuIFN-λ1-PMD18-T Vector和HuIFN-λ2-PMD18-T Vector的SacⅠ、XhoⅠ和SacⅠ和SalⅠ双酶切鉴定, 均出现610 bp大小的带, 测序示与GenBank上报道的序列完全一致.PCAGG-HuIFN-λ1活性104 IU/mL, PCAGG-HuIFN-λ2活性为102 IU/mL, 且PCAGG-HuIFN-λ1诱导Mx抗病毒蛋白的表达一定时间内随时间延长表达不断增强, 9～12 h达高峰, 24 h后消失(P＜0.05).结论:成功构建了PCAGG-HuIFN-λ1PCAGG-HuIFN-λ2 的真核表达载体并在BHK-21细胞中得到表达, 其抗病毒活性与诱导Mx抗病毒蛋白密切相关.     

一例促皮质素静脉外渗至局部皮肤坏死的案例解析与改进

对1例癫痫性痉挛患儿采用促皮质素静脉给药时发生静脉外渗至局部皮肤坏死的案例进行解析。针对事件发生原因,制定并采取措施:静脉穿刺难度事前评估,优化临床监护,加强相关知识培训,以肌肉注射代替静脉滴注等,避免了此类事件的再次发生,提升了临床合理用药水平,确保了患者安全。     

胞磷胆碱胶囊治疗血管性认知障碍患者的疗效及对认知功能的影响

目的:研究胞磷胆碱胶囊治疗血管性认知障碍(VCI)患者的疗效及对认知功能的影响。方法:选取2016年1月至2021年12月郑州市中心医院收治的80例VCI患者作为研究对象,随机分为对照组和观察组,各40例。在口服抗血小板药物阿司匹林预防心血管事件和脑血管事件,口服降脂药物阿托伐他汀预防心血管事件,行生活方式干预、认知训练、心理治疗,缓解情绪问题和改善认知功能等基础上,对照组患者口服安慰剂治疗,观察组患者口服胞磷胆碱胶囊治疗。比较两组患者治疗效果、认知功能情况及生活质量。结果:观察组患者患者治疗有效率高于对照组,差异具有统计学意义(P <0.05)。治疗8周及16周,观察组患者认知功能评分、生活质量评分高于对照组,差异具有统计学意义(P <0.05)。结论:口服胞磷胆碱胶囊可以有效改善VCI患者的认知功能,提高治疗效果,改善患者生活质量。     

腹腔镜针对复杂性胆囊及胆总管结石术中的临床决策及处理技巧

目的 探讨腹腔镜术中处理复杂性胆囊结石及胆总管结石的不同方式对手术的影响。方法回顾性分析青岛市市立医院收治的复杂性胆囊及胆总管结石患者136例,总结并归纳术中情况及相对应处理技巧及经验。结果 本研究中腹腔粘连病例占全组120例（88.2%）,其中炎症粘连73例,手术后粘连47例,术中遵循“宁伤胆囊、远离肝门”的处理原则及相应的处理技巧,98例（81.7%）腹腔粘连患者按照原定手术方式顺利完成,但22例（18.3%）腹腔粘连患者因术中胆总管或胆囊颈管辨别不清、术中发现胆囊癌变等原因施行中转开腹手术。结石嵌顿病例占全组16例（11.8%）,包括胆囊颈管及胆总管结石嵌顿,术中应用推挤法、直接取石法、液电碎石等技巧联合应用胆道镜、碎石仪及扩张球囊等设备,手术均在腹腔镜下完成。结论 腹腔粘连和结石嵌顿是腹腔镜下复杂性胆囊结石及胆总管结石手术中较常见的困难,相应的术中决策和处理技巧能够为提高患者安全、降低手术风险、减少术后并发症等提供更多的方法与思路。     

热化疗对人体结肠癌LoVo细胞中一氧化氮含量的影响

Aim To study the effect of nitric oxide (NO) on LoVo cells after hyperthermia combined with MMC and their mechanisms. Methods The levels of NO were determined in cell culture medium of LoVo cells by Cu-Cd reduction method.Results The levels of NO in control (45.9±16.12) μmol/L group were lower than those in chemohyperthermy(92.3±24.30) μmol/L and hyperthermia (67.67±15.48)μmol/L group (P＜0.01,P＜0.05 respectively). Levels of NO in chemohyperthermy and hyperthermia groups were higher than those in chemotherapy(47.03±16.32)μmol/L(P＜0.01,P＜0.05 respcevtively). The levels of NO in chemo-hyperthermy group were higher than those in hyperthermia group(P＜0.05), and those in chemotherapy group higher than in control group but without statistical significance.Conclusion Chemohyperthermy and hyperthermia could remarkably enhance the levels of NO produced by LoVo cells. NO levels could also be enhanced by chemotherapy. It could be concluded that Nitric Oxide Synthase (NOS) in tumor cells might be activated by hyperthermia, chemohyperthermy and chemotherapy, and increased NO level which induced cytotoxic reaction. This may be one of the mechanisms of treatment with chemohyperthermy.