Small envelope protein E of SARS：cloning,expression, purification, CD determination, and bioinformatics analysis

AIM:To obtain the pure sample of SARS small envelope E protein (SARS E protein), study its properties and analyze its possible functions. METHODS: The plasmid of SARS E protein was constructed by the polymerase chain reaction (PCR), and the protein was expressed in the E coli strain. The secondary structure feature of the protein was determined by circular dichroism (CD) technique. The possible functions of this protein were annotated by bioinformatics methods, and its possible three-dimensional model was constructed by molecular modeling. RESULTS: The pure sample of SARS E protein was obtained. The secondary structure feature derived from CD determination is similar to that from the secondary structure prediction. Bioinformatics analysis indicated that the key residues of SARS E protein were much conserved compared to the E proteins of other coronaviruses. In particular, the primary amino acid sequence of SARS E protien is much more similar to that of murine hepatitis virus(MHV) and other mammal coronaviruses. The transmembrane (TM) segment of the SARS E protein is relatively more conserved in the whole protein than other regions. CONCLUSION: The success of expressing the SARS E protein is a good starting point for investigating the structure and functions of this protein and SARS coronavirus itself as well. The SARS E protein may fold in water solution in a similar way as it in membrane-water mixed environment. It is possible that β-sheet I of the SARS E protein interacts with the membrane surface via hydrogen bonding, this β-sheet may uncoil to a random structure in water solution.     

Tanshinone IIA triggers p53 responses and apoptosis by RNA polymerase II upon DNA minor groove binding

Our previous work has shown that tanshinone IIA (Tan IIA) is a DNA minor groove binder instead of an intercalator as previously thought. In this study, we have further demonstrated that the molecular antitumor pharmacology of Tan IIA is dependent on its groove-binding capability. First, we investigated the structure damage to duplex DNA upon Tan IIA binding using circular dichroism spectra. Subsequently, we performed western blot, flow cytometry analysis, chromatin immunoprecipitation, and quantitative real-time PCR to illustrate the RNAPII degradation, phosphorylation, and distribution along the transcribed gene in H22 cells exposed to Tan IIA. In addition, p53 activation and apoptosis induction in both cultured H22 cells and in mice bearing the ascitic-type H22 were measured following Tan IIA treatment. It was revealed that Tan IIA decreases the level of RNAPII by altering DNA structure. At the low dose range (0.2-4 μM) of Tan IIA exposure, the DNA structure damage results in the inhibition of RNAPII binding to DNA and the initiation of RNAPII phosphorylation, while higher concentrations of Tan IIA (4-20 μM) cause complete phosphorylation and degradation of RNAPII followed by p53 activation and apoptosis. A similar apoptosis induction by RNAPII was observed in animals. Apoptosis of tumor cells from ascitic fluid was not detected until RNAPII levels were downregulated by Tan IIA, which requires 40 mg/kg body weight of Tan IIA. It was concluded that DNA-conformational-damage-dependent RNAPII response upon groove binding is the molecular basis of the antitumor property of Tan IIA, in vivo and in vitro.     

siRNA沉默卵巢癌CXCR4基因对卵巢癌腹腔转移的影响

目的研究应用基因靶向治疗对卵巢癌腹腔转移的实验疗效观察。方法制作好的卵巢癌大鼠模型60只,采用随机数字表法随机分为实验组和对照组,实验组应用基因靶向治疗,对照组采用阴性治疗,siRNA沉默卵巢癌CXCR4基因后,观察CXCR4蛋白的表达情况,对卵巢癌细胞增殖的影响,观察应用基因靶向治疗后腹腔转移情况,评价大鼠生存情况。结果实验组在治疗后腹水中的CXCR4显著减少（P〈0．05）;对照组治疗后腹水中的CXCR4量显著性增加（P〈0．05）;实验组与对照组相比,治疗前腹水中CXCR4,差异无统计学意义（P〉0．05）;治疗后差异有统计学意义（P〈0．05）。实验组腹水抽出量与实验组相比,明显减少（P〈0．05）。结论应用基因靶向治疗方法siRNA沉默卵巢癌CXCR4基因对卵巢癌腹腔转移,具有较好的疗效,临床预后较好。     

普罗帕酮终止室上性心动过速

目的 通过对普罗帕酮终止室上性心动过速时不同剂量、给药速度与疗效、不良反应的观察,探讨其最佳给药方法与剂量.方法 随机将符合入选标准的246例患者分为A、B、C、D四组.A、C组分别以每千克体重1mg、2mg的剂量静脉推注,1min完毕;B、D组分别在A、C组的基础上加5%葡萄糖10ml稀释,静脉推注5min完毕.观察各组间转复率、转复时间、不良反应发生率.结果 ①给药速度越快,转复为窦性心律越快,不良反应越小.②随剂量的增加,不良反应发生率大幅度增加;而转复成功率、转复时间无显著性差异.结论 以每千克体重1mg的普罗帕酮1min静注完毕,能快速终止室上性心动过速的发生且不良反应发生率最低.     

社会支持对精神分裂症患者生活质量和康复效果的影响

目的 探讨社会支持对精神分裂症患者生活质量和康复效果的影响.方法 选取华北理工大学附属开滦精神卫生中心于2014年2月-2015年8月收治的慢性精神分裂症278例,按照干预方法分成观察组和对照组,每组139例.对照组给予常规康复干预,观察组在对照组基础上实施社会支持模式干预.采用社会支持评定量表(SSRS)、精神分裂症患者生活质量量表(SQLS)评分和简明精神病评定量表(BPRS)评估干预后患者的生活质量和康复效果.结果 观察组干预后SSRS评分高于干预前和对照组,SQLS和BPRS评分均低于干预前和对照组(P＜0.05).观察组SQLS总均分与SSRS中主观支持、客观支持和总均分呈负相关,BPRS总均分与SSRS中客观支持、支持利用度和总均分呈负相关(P＜0.05).结论 社会支持能明显改善精神分裂症患者生活质量和康复水平,应重视社会支持对精神分裂症预后的影响.     

大鼠肠道菌群对长梗冬青苷体外代谢的影响

摘 要 目的：探究体外孵育的大鼠肠道菌群对长梗冬青苷的代谢转化规律。方法: 将长梗冬青苷与大鼠肠道菌群在厌氧条件下共孵育,分别于0,4,8,12,24,48 h时取样,经乙酸乙酯萃取后采用HPLC法进行定性和定量分析。结果: 在与大鼠肠道菌群共孵育48 h后,90.8%的长梗冬青苷被代谢转化为M₂,M₂与铁冬青酸对照品通过HPLC分析时色谱行为一致,故确定M₂为铁冬青酸。结论:长梗冬青苷可以被体外孵育的大鼠肠道菌群代谢转化为铁冬青酸。     

2012 年我院住院患者金黄色葡萄球菌感染调查及耐药性分 析简

目的 对2012年医院住院患者金黄色葡萄球菌感染现状及耐药性进行调查,为临床用药提供参考.方法 回顾性分析2012年住院患者细菌培养标本中金黄色葡萄球菌的分离情况及耐药性.结果 共分离金黄色葡萄球菌305株,其中耐甲氧西林金黄色葡萄球菌（MRSA）191株,占62.62％.金黄色葡萄球菌感染的标本来源主要有痰液（44.92％）、伤口分泌物（24.92％）和脓液（11.15％）;感染科室主要以骨外科、重症监护室（ICU）、神经外科、老年病房和呼吸内科最多见.药物敏感性试验结果显示,MRSA耐药率明显高于甲氧西林敏感金黄色葡萄球菌（MSSA）,未发现对万古霉素、替考拉宁、利奈唑胺及替加环素耐药的菌株,克林霉素诱导耐药率为36.36％.结论 金黄色葡萄球菌耐药严重,临床应根据药物敏感性试验结果合理使用抗菌药物.     

曲美他嗪治疗慢性心力衰竭疗效观察

目的观察曲美他嗪(商品名为万爽力)对慢性心力衰竭的治疗效果。方法根据病史、查体、相关辅助检查及心脏彩色多普勒超声检查结果,并参照《慢性心力衰竭诊断治疗指南》及纽约心脏病协会(NYHA)心功能分级标准,选择72例NYHA心功能分级Ⅱ-Ⅳ级慢性心力衰竭患者,随机分为治疗组和对照组各36例。治疗组在常规治疗基础上加万爽力口服,每次20 mg,每天3次;对照组给予常规治疗,疗程均为3个月,治疗结束评价万爽力治疗慢性心衰竭的疗效。结果治疗组心脏彩色多普勒超声检查左心室收缩末期内径(LVESD)、左心室舒张末期内径(LVEDD)、左心室射血分数(LVEF%)等指标有所改善,临床主要症状及体征、心功能等改善情况均明显优于对照组(P<0.05)。结论万爽力治疗慢性心力衰竭疗效较好,患者依从性好,未见明显不良反应,值得临床推广应用。     

Neural representation of phonological information during Chinese character reading

Previous studies have revealed that phonological processing of Chinese characters elicited activation in the left prefrontal cortex, bilateral parietal cortex, and occipitotemporal regions. However, it is controversial what role the left middle frontal gyrus plays in Chinese character reading, and whether the core regions (e.g., the left superior temporal gyrus and supramarginal gyrus) for phonological processing of alphabetic languages are also involved in Chinese character reading. To address these questions, the present study used both univariate and multivariate analysis (i.e., representational similarity analysis, RSA) to explore neural representations of phonological information during Chinese character reading. Participants were scanned while performing a reading aloud task. Univariate activation analysis revealed a widely distributed network for word reading, including the bilateral inferior frontal gyrus, middle frontal gyrus, lateral temporal cortex, and occipitotemporal cortex. More importantly, RSA showed that the left prefrontal (i.e., the left middle frontal gyrus and left inferior frontal gyrus) and bilateral occipitotemporal areas (i.e., the left inferior and middle temporal gyrus and bilateral fusiform gyrus) represented phonological information of Chinese characters. These results confirmed the importance of the left middle frontal gyrus and regions in ventral pathway in representing phonological information of Chinese characters.