基于DNA条形码和HRM技术建立紫草药材的RFLP-HRM鉴别方法

目的:基于DNA条形码技术和高分辨率熔解曲线(HRM)技术鉴别紫草市场样品的真伪并建立紫草药材的RFLP-HRM鉴别方法。方法:通过聚合酶链式反应(PCR)对紫草药材的ITS2序列进行扩增;利用MEGA 6.06软件对紫草药材的ITS2序列进行NJ树聚类分析和遗传距离分析;对市场中紫草非药典品进行基原确证;针对紫草药材的ITS2序列利用Primer Premier 5软件进行引物设计;对经设计、合成的引物通过HRM实验进行筛选;建立紫草药材的RFLP-HRM鉴定方法。结果:通过对紫草药材的ITS2序列进行NJ树聚类分析和遗传距离分析,可以有效区分新疆紫草、疏花软紫草、黄花软紫草及市场上非药典来源的紫草药材(紫草非药典品);经过Primer Premier 5软件进行引物设计共选出4对引物进行合成;通过HRM实验筛选出引物Zcf-1为最佳实验引物;确立了有效鉴别紫草市场药材的RFLP-HRM方法。结论:通过DNA条形码和HRM技术均能有效鉴别紫草市场药材真伪,且结果一致,起到互相验证的作用,建立了紫草市场药材的RFLP-HRM鉴定方法,为紫草及其他中药民族药品种的快速鉴定技术提供了可以参...     

有经验术者行非体外循环与体外循环冠状动脉旁路移植术的近期安全性比较：单中心31 075例手术对比研究

目的:比较在单一高手术量心脏外科中心,有经验术者行体外循环和非体外循环冠状动脉旁路移植术（coronary artery bypass graft,CABG）的近期安全性结果。方法:本研究为单中心回顾性研究,连续选取2009年1月至2019年12月共31 075例在阜外医院由有经验术者（既往CABG完成超过100例）完...     

说说蛋黄的颜色

正许多人以蛋黄颜色来判断土鸡蛋与洋鸡蛋。其实这是很不准确的。蛋黄的黄色可从很浅到很深,食品行业通常用罗氏蛋黄颜色扇(RYCF)来确定蛋黄的色度,颜色越深,数字越高,最高为15。蛋黄的颜色,     

丹参注射液治疗普外科手术后患者下肢深静脉血栓形成的临床观察研究

目的研究丹参注射液预防性治疗普外科手术后患者下肢深静脉血栓形成的临床效果。方法我院普外科2016年7月到2018年6月期间实施手术治疗的患者120例,随机分为对照组和试验组各60例。对照组实施常规手术治疗,试验组患者采用丹参注射液进行预防性治疗,比较两组术后并发症发生率。结果试验组患者术后并发症发生率低于对照组,数据对比P0.05。试验组的凝血酶原时间为(12.49±0.62)s、活化部分凝血活酶时间为(27.46±3.29)s,比对照组的(14.28±0.85)s、(31.46±5.71)s短,数据对比P0.05。试验组的纤维蛋白原水平为(3.26±0.41)g/L,比对照组的(4.32±0.57)g/L低,数据对比P0.05。结论丹参注射液在普外科手术后患者下肢深静脉血栓形成中的预防应用效果较好,能有效降低患者术后下肢深静脉血栓等并发症,改善凝血酶原时间与纤维蛋白原水平等指标,具有较高临床价值。     

阴道超声对卵巢癌淋巴结转移的诊断效能分析

目的:探讨阴道超声对卵巢癌淋巴结转移的诊断效能分析.方法:入选2017年6月～2020年6月某院86例卵巢癌患者作为研究对象,所有患者均使用超声检查,记录超声检查结果和患者淋巴结转移的情况,并对其进行分析.结果:阴道超声对卵巢癌淋巴结转移阳性有55例,阴性有31例.手术判断阳性有56例,阴性30例,共误诊1例,漏诊2例...     

柯萨奇病毒B3 VP1蛋白、rAd/VP1和pcDNA3/VP1的免疫效果比较

目的 观察柯萨奇病毒B3(Coxsackievirus B3,CVB3)衣壳蛋白VP1、表达VP1蛋白的重组腺病毒rAd/VP1和重组质粒pcDNA3/VP1的免疫效果.方法 用原核细胞表达VP1蛋白并纯化、扩增重组腺病毒rAd/VP1,扩增并提取真核表达质粒pcDNA3/VP1.BALB/c小鼠随机分为4组,每组18只,分别在股四头肌注射VP1蛋白、rAd/VP1、pcDNA3/VP1和PBS.VP1蛋白组和pcDNA3/VP1组免疫3次,间隔3周;rAd/VP1组免疫2次,间隔2周.VP1蛋白、pcDNA3/VP1和rAd/VP1每次每只注射剂量分别为50μg、100μg和1.2×107PFU.用ELISA法和微量中和试验法检测各次免疫后血清CVB3特异性IgG抗体和中和抗体滴度;末次免疫后3周,CCK-8法检测脾脏淋巴细胞的CTL杀伤活性;用致死量CVB3攻击小鼠后,检测血中病毒滴度并观察动物的存活情况.结果 VP1蛋白组血清特异性IgG抗体和中和抗体滴度明显高于其他实验组(P＜0.05),而脾脏淋巴细胞CTL杀伤活性低于rAd/VP1组(P＜0.05);致死量病毒攻击后,VP1蛋白组血中病毒滴度低于pcDNA3/VP1和rAd/VP1组(P＜0.05),生存率明显高于这两组(P＜0.05).结论 VP1蛋白疫苗能诱导较高水平的体液免疫应答,对动物有明显的免疫保护作用,免疫效果优于质粒pcDNA3/VP1和重组腺病毒rAd/VP1.

Abstract：

Objective To compare the immune effects of Coxsackievirus B3 (CVB3) capsid protein VP1 expressed bacterially, recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1which express VP1 protein in mice. Methods After expressed in prokaryotic cells, VP1 protein was purified. Recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1 were amplified and extracted. Six to 8-week-old, male BALB/c mice were divided into four groups randomly. Each group contained 18 mice. The mice of pcDNA3/VP1 group or VP1 protein group were immunized intramuscularly with three injections at three weeks apart, of recombinant plasmid pcDNA3/VP1 at a dose of 100 μg/mouse or recombinant protein VP1 at a dose of 50 μg/mouse. The mice of rAd/VP1 group were immunized intramuscularly twice at two weeks interval with rAd/VP1 at a dose of 1.2 × 107 PFU. The control group was mock-immunized with 100 μl of PBS intramuscularly. Mice were bled from the retroorbital sinus plexus every two weeks after each immunization. ELISA and micro-neutralization test were used to detect levels of CVB3-specific IgG antibody and neutralizing antibody titers in the sera of immunized mice. Three weeks after the last immunization, the cytotoxic T lymphocyte(CTL) killing activity of spleen lymphocytes was detected with CCK-8 assay. Subsequently, virus titers in the sera of immunized mice were determined by the 50% cell culture infective dose( CCID50 ) assay on HeLa cell monolayers and percentage of animals surviving were observed after lethal CVB3 attack over a period of 21 days. Results The titers of specific IgG antibody and neutralizing antibody in sera of VP1 protein immunized mice were higher than other groups( P ＜0.05 ). While CTL killing activity of spleen lymphocytes of VP1 protein immunized mice was lower than mice in rAd/VP1 group( P ＜0. 05). Virus titers in sera of VP1 protein immunized mice were lower than the mice in pcDNA3/VP1 or rAd/VP1 groups ( P ＜ 0.05 ), while survival rate was significantly higher than these two groups ( P ＜ 0.05 ).Conclusion VP1 protein induced higher level of humoral immune response and acquired obvious immune protection effects in mice. The immunizing potency of VP1 protein vaccine surpassed plasmid pcDNA3/VP1or recombinant adenovirus rAd/VP1. It appeared to be a promising candidate among the three different vaccines.     

慢性疼痛与器质性疼痛心理学特征的对照研究

目的 旨在探讨慢性疼痛与器质性疼痛心理学特征的差异。方法 采用ＳＣＬ－９０、ＬＥＳ、ＳＳＲＳ、ＤＳＱ、ＨＡＭＤ、ＨＡＭＡ等尽理学量表对５９例慢性疼痛（ＣＰ）与５０例器质性疼痛（ＯＰ）患者进行对照研究,并用ｔ检验对数据进行统计分析。结果 与ＯＰ组相比较,ＣＰ组有较多的生活支持相对缺乏,较易动用不成熟防御方式,有明显的抑郁、焦虑情绪。结论 慢性疼痛与心理社会因素密切相关。