用多重及套式PCR同时检测血清中的HBV和HCV方法的建立

首次应用多重及套式ＰＣＲ技术同时检测人血清中的ＨＢＶ和ＨＣＶ。将抽提的ＨＢＶＤＮＡ和（或）ＨＣＶＲＮＡ在含ＡＭＶ逆转录酶、ＴａｑＤＮＡ多聚酶以及ＨＢＶ和ＨＣＶ外套引物的ＰＣＲ缓冲引物的ＰＣＲ缓冲液中进行逆转录后连续进行ＰＣＲ扩增，以第一轮扩增产物为模板，在含ＨＢＶ和ＨＣＶ内套引物的ＰＣＲ反应体系中进行第二轮扩增，产物经电泳后，以ＤＮＡ分子量标志物或已知片段做参考，出现５２３ｂｐ和（或）２６０ｂｐ产     

丙型肝炎患者血清和外周血单个核细胞中HCVRNA检测及其临床意义

应用套式ＰＣＲ检测了８８例丙型肝炎患者血清和外周血单个核细胞（ＰＢＭＣ）中ＨＣＶＲＮＡ；结果发现：慢性丙肝患者ＰＢＭＣ中ＨＣＶＲＮＡ检出率显著高于急性丙肝患者（Ｐ＜０．００１）；急、慢性丙肝患者血清和慢性丙肝患者ＰＢＭＣ中ＨＣＶＲＮＡ检出率显著高于ＡＬＴ正常的抗－ＨＣＶ阳性者（Ｐ＜０．００１）；少数患者血清中ＨＣＶＲＮＡ阴性，ＰＢＭＣ中可测及，部分患者血清抗一ＨＣＶ阴性，而血清ＨＣＶＲＮＡ阳性．提示：血清ＨＣＶＲＮＡ检测有助于抗－ＨＣＶ阴性丙肝的诊断和早期诊断；丙肝的肝损害可能与ＨＣＶＲＮＡ血症有关；ＰＢＭＣ中ＨＣＶ感染在丙肝的慢性化和慢性肝损害中可能起一定作用；ＰＢＭＣ中可贮存ＨＣＶＲＮＡ。     

应用原位聚合酶链反应对肝细胞癌组织中HBVDNA进行再评价

为探讨ＨＢＶ感染与肝细胞癌（ＨＣＣ）发生之间关系，作者应用原位聚合酶链反应（ＰＣＲＩＳ）对２６例肝癌、２３例癌旁肝组织的石蜡切片进行了ＨＢＶＤＮＡ的检测，检出率分别为８４．６％和９１．３％，明显高于原位核酸杂交（ＩＳＨ）的５３．８％（１４／２６）和５６．５％（１３／２３）；亦高于从患者血清和ＨＣＣ组织提取ＤＮＡ后行聚合酶链反应（ＰＣＲ）扩增的５７．７％（１５／２６）和５５．６％（５／９）。用ＰＣＲＩＳ方法，ＨＣＣ组织细胞中ＨＢＶＤＮＡ阳性颗粒强度、ＨＢＶＤＮＡ阳性细胞数和组织切片的清晰度均高于ＩＳＨ。ＰＣＲＩＳ既显示了ＰＣＲ技术的高敏性和特异性，又能在细胞内定位。本结果显示我国ＨＣＣ的发生与ＨＢＶ感染密切相关。     

家庭内乙型肝炎病毒变异株感染的分子病毒学调查

为分析乙型肝炎病毒（ＨＢＶ）家庭内聚集感染病例的ＨＢＶ变异特点，我们对１０个家庭２５例ＨＢＶ感染者进行了分子病毒学研究，均用单链构型多态性分析（ＳＳＣＰ）ＨＢＶ外膜基因“ａ”决定簇片段的变异，ＰＣＲ－限制性片断长度多态性分析，分析ＨＢＶ前Ｃ区终码变异。结果与用聚合酶链反应（ＰＣＲ）产物直接序列分析测定的４个家庭有、无ＨＢＶ变异株感染的外膜基因序列和前Ｃ基因序列比较，从分子水平鉴定ＨＢＶ家庭内传播，同时建立了ＳＳＣＰ检测ＨＢＶ免疫逃避变异株的方法。     

HBV—DNA套式一次PCR方法的建立

应用乙型肝炎病毒ＤＮＡ Ｃ基因区的一对外引物（ＨＢＣ１和ＨＢＣ２，护增片段长度为５６６ ｂｐ）及一对内引物（ＨＢＣ３和ＨＢＣ４，护增片段长度为３０１ ｂｐ）对不同稀释度的ＨＢＶ－ＤＮＡ（ａｄｒ亚型）进行套式一次ＰＣＲ扩增，结果表明灵敏度可达到１０ａｇ水平。此方法快速、简便、灵敏、特异，与分子杂交的灵敏度相当。     

输血传播病毒(TTV)感染的肝病患者36例分析

目的：观察ＴＴＶ（Ｔｒａｎｓｆｕｓｉｏｎ ｔｒａｎｓｍｉｔｔｅｄｖｉｒｕｓ,ＴＴＶ）阳性肝病患者的临床特征。方法：用巢式ＰＣＲ法检测血清ＴＴＶ ＤＮＡ,同时观察血清生化指标和肝组织学的变化。结果：ＴＴＶ感染者的血清学模式以ＨＢＶ、ＨＣＶ、ＨＡＶ、ＨＥＶ混合感染为主,占６１．１％（２２／３６）,临床表现以乏力、纳关为特征,ＴＢＬ、ＡＬＴ变化不明显;而ＴＴＶ单独感染者为３８．９％（１４／３６）,临床特     

烧伤病人巨细胞病毒感染的检测分析

目的：观察严重烧伤病人巨细胞病毒（ＨＣＭＶ）感染情况及对病情的影响。方法：采用ＰＣＲ技术６０例烧伤病人创面及尿液中ＨＣＭＶ－ＤＮＡ,用ＥＬＩＳ观察血清中ＨＣＮＭＶ－ＩｇＭ、ＩｇＧ,并与５０例正常检测结果进行对照。结果：观察组血清中ＨＣＭＶ－ｇＭ和创面及尿液中ＨＣＭＶ－ＤＮＡ均高于对照组,感染ＨＣＭＶ的烧伤病人创而愈合较慢,外周血中淋巴细胞计数较高。结论：烧伤患感染ＨＣＭＶ的机会较大,且烧伤后早     

PCT检测HCV-RNA阳性与血清HCV抗体的对比研究

ＰＣＴ检测ＨＣＶ－ＲＮＡ阳性与血清ＨＣＶ抗体的对比研究武警河南总队医院病理科梁喜林，郑晓芙，彭亚丁，粟娟（郑州４５００５２）关键词ＰＣＲ检测，ＨＣＶ－ＲＮＡ，抗－ＨＣＶ采用ＰＣＲ技术检测ＨＣＶ－ＲＮＡ及酶免检测血清丙型肝炎抗体（抗－ＨＣＶ）来测定及辅...     

聚合酶链反应检测HBV感染患者血清HBV DNA的临床意义

聚合酶链反应（Ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ，ＰＣＲ）技术是本世纪分子生物学重大进展之一，在体外可将特异ＤＮＡ序列呈百万倍扩增，具有高灵敏度和高特异性。随着该技术的不断改进和完善，其在ＨＢＶ感染研究中的应用也越来越多，大大丰富和更新了人们对ＨＢＶ感染的认识［１］。本文应用ＰＣＲ技术检测不同ＨＢＶ感染患者血清中ＨＢＶＤＮＡ，旨在探讨急、慢性乙型肝炎及与ＨＢＶ感染相关的肝硬变和肝癌患者血清ＨＢＶＤＮＡ的存在情况及其与乙型肝炎病毒血清标志物（ＨＢＶＭ）在反映体内病毒复制方面存在的差异，…     

HBV－DNA套式一次PCR方法的建立

应用乙型肝炎病毒ＤＮＡＣ基因区的一对外引物（ＨＢＣ＿１和ＨＢＣ＿２，扩增片段长度为５６６ｂｐ）及一对内引物（ＨＢＣ＿３和ＨＢＣ＿４，扩增片段长度为３０１ｂｐ）对不同稀释度的ＨＢＶ－ＤＮＡ（ａｄｒ亚型）进行套式一次ＰＣＲ扩增，结果表明灵敏度可达到１０ａｇ水平。此方法快速、简便、灵敏、材异，与分子杂交的灵敏度相当。     

Physiologic and pathologic calcifications and ossifications in the face and neck

The aim was to give a systematic presentation of physiologic and pathologic calcifications and ossifications in the face and neck with a special emphasis on clinical relevance. In a sometimes subacute setting one should recognize specific calcifications which often lead to important diagnoses such as fungal sinusitis or sclerosing labyrinthitis. In a more chronic situation intraocular calcifications in small children are pathognomonic for retinoblastoma. Juxtatumoral sclerosis of the laryngeal cartilage in laryngopharyngeal carcinoma is usually caused by tumor infiltration of the cartilage resulting in a higher tumor stage and, this way, has a major impact on the therapeutical strategy. Calcified lymph nodes are mainly unspecific but can be the result of tuberculosis or metastases of thyroid cancer. Cross-sectional imaging methods, most of all computed tomography, are ideally suited to reveal head and neck calcifications and ossifications, especially those which are clinically relevant.     

Head and neck infection and inflammation

This article discusses the imaging manifestations of infectious and inflammatory conditions of the head and neck. Special attention is paid to the sites, routes of spread, and complications of neck infections. Because the clinical signs and symptoms and the complications of these conditions are often determined by the precise anatomic site involved, anatomic considerations are stressed. Familiarity with the fascial layers, spaces of the neck, and the contents of each space is helpful for this discussion. The fascial layers of the neck are important barriers to infection, and once infection is established, the fascial layers play a part in directing its spread.     

幽门螺杆菌分泌与重组表达的VacA蛋白的分离纯化及其致细胞空泡效应与凋亡的对比研究

目的：分离纯化幽门螺杆菌分泌和重组表达的细胞空泡毒素抗原（ VacA）蛋白,并评价其致细胞空泡效应及致细胞凋亡效应。方法分别从幽门螺杆菌ATCC26695菌株培养上清和重组表达VacA蛋白的pQE30-VacA-E．coliM15基因工程菌中分离纯化VacA蛋白,经酸化后,以不同终浓度（5,10 ng／ml）分别与人胃腺癌AGS细胞共孵24 h,观察致空泡效应,并通过流式细胞术检测细胞凋亡。结果成功分离纯化出幽门螺杆菌分泌和重组表达的VacA蛋白;幽门螺杆菌分泌的VacA蛋白能显著引起AGS细胞的空泡样改变及凋亡（P＜0．01）,而重组表达的VacA蛋白致细胞空泡样改变及凋亡不显著（ P＞0．05）。结论幽门螺杆菌分泌的VacA蛋白有良好的空泡毒性及致凋亡效应,而重组表达的VacA蛋白无致空泡及凋亡效应,幽门螺杆菌分泌的VacA蛋白可用于VacA作用机制的研究。     

化学和生物学融合对国际武器公约履约的影响

化学武器公约（ CWC）和生物武器公约（ BWC）是为禁止生产、发展、储存和使用化学武器和生物武器而制定的国际公约。近年来,科学技术快速发展,知识交叉渗透,学科之间出现整合和融合,促进了科技进步和经济发展。其中化学和生物学融合在有力促进制药、健康卫生、绿色化学和环境保护等产业进步的同时,也对化学和生物武器公约的履约产生了重要的影响。该文综述了与化学武器和生物武器公约相关的化学和生物学融合进展,并分析其对公约履约的影响。     

Oxygen uptake and ventilation during rowing and running in females and males

This study evaluated if the ventilatory response to exercise is impaired by the cramp position of rowing. Maximal oxygen uptake (VO2max), maximal expiratory volume (VEmax), and maximal heart rate (HRmax) during rowing and running were compared in 55 males (age, mean +/- SD, 21 +/- 3 years; height 176 +/- 5 cm; body mass 72 +/- 6 kg) and 18 females (age 20 +/- 2 years; height 164 +/- 5 cm; body mass 61 +/- 4 kg). VEmax was larger during rowing than during running (males, 157 +/- 16 vs. 147 +/- 13 L min(-1); 114 +/- 9 vs. 105 +/- 11 L min(-1), P<0.01). Also VO2max was larger during rowing than during running (males, 4.5 +/- 0.5 vs. 4.3 +/- 0.4 L min(-1); females, 3.3 +/- 0.4 vs. 3.2 +/- 0.4 L min(-1), P<0.01). However, HRmax was lower during rowing than during running (males, 194 +/- 8 vs. 198 +/- 11 beats min(-1); females, 192 +/- 6 vs. 196 +/- 8 beats min(-1), P<0.05). VEmax was correlated to body mass and fat-free mass, as was VO2max. Thus, the oxygen pulse (VO2max/HRmax) was larger during rowing than during running, while the ventilatory equivalent for oxygen (VEmax/VO2max) was similar. We showed that bending the body during rowing does not seem to impair ventilation either in males or in females. The results indicate that VEmax and VO2max relate to body size and fat-free mass for both females and males. The findings indicate that the involvement of more muscles, the entrainment, and the body position during rowing facilitates ventilation and venous return and lowers maximal heart rate.     

Coracoclavicular and coracoacromial ligament calcification and ossification

Thirty-six patients with calcification or ossification at or around the coracoclavicular and coracoacromial regions were analyzed with regard to type, location, and configuration of the deposits and related clinical history. Calcification or ossification in the coracoclavicular region resulted largely from trauma (36%) or renal failure (28%). Trauma patients may develop punctate calcification or ossification but do not develop the tumoral type of calcification. About 5% of the renal failure patients had coracoclavicular ligament calcifications, one-half of which were of the tumoral type. Renal failure patients may have punctate or tumoral calcifications but do not develop ossification.     

CLIP相关技术的研究进展

在真核生物基因表达的转录后调节中,RNA结合蛋白（ RBP）起着关键作用,很多RBP的异常与人类疾病的发生密切相关。自2000年的RNA免疫沉淀和芯片分析方法（ RNA immunoprecipitation with differential display or microarray analysis , RIP-ChIP）出现以来,人们开始就RBP与RNA相互作用进行了系统而广泛的研究。经过改良和发展,基于体内实时紫外交联免疫沉淀法（ ultraviolet crosslinking and immunoprecipitation , CLIP ）、交联免疫沉淀cDNA文库高通量测序法（ high-throughput sequencing of CLIP cDNA library , HITS-CLIP）、光催化核糖核苷增强交联和免疫沉淀法（ photoactivatable-ribonucleoside-enhanced crosslinking and immunprecipitation , PAR-CLIP）以及提高个别核苷酸分辨率交联和免疫共沉淀法（ individual nucleotide resolution CLIP , iCLIP）等RIP-ChIP衍生方法相继产生,使用这些方法,可以解析RBP的RNA识别特异性,而且通过与高通量测序技术结合,可以实现转录组尺度的RBP的靶序列的鉴定,分辨率也得到极大提高。该文就RNA与蛋白的相互作用的基本原理及其研究进展、相关技术存在的问题以及发展趋势进行简要综述。     

Old and dangerous: Prison and dementia

Older prisoners are the fastest growing group of prisoners in many countries. The purpose of this study is to explore the phenomenon of detention of persons suffering from dementia. Medline searches were conducted for relevant articles, chapters and books published until August 2016. Search terms included dementia, elderly, prison and criminal. Publications found through this indexed search were reviewed for further relevant references. As results, there is a lack of data about elderly with dementia in prisons. Given the rise in the average age, it is reasonable to hypothesize that the number of older prisoners is growing. Moreover, some elderly are imprisoned with a concomitant cognitive impairment or psychiatric disorder while others will develop such diseases once incarcerated. At the present time, legal and social systems seem unprepared to handle the phenomenon of dementia in prison. As proposal, health assessments for older first time offenders should become a practice inside the correctional facilities and include an evaluation for specific health issues, such as psychiatric comorbidity and cognitive impairment.     

Kidney and iodinated and gadolinium-based contrast agents

In patients with renal failure, iodinated contrast agents may cause acute deterioration of the renal function and gadolinium-based contrast agents (GBCAs) may cause nephrogenic systemic fibrosis (NSF). The administration of a contrast agent must thus be reviewed for each patient and evaluation of renal function is paramount even though its estimation using formulas derived from the creatinine level may fluctuate. For iodinated contrast agents, contrast induced nephropathy is reduced by hydratation, preferably intravenous, when the GFR is less than 60 ml/min. The risk for intravenous injections is less than the risk for arterial injections, and the GFR threshold may be reduced to 45 ml/min. For gadolinium-based contrast agents, patients at risk for NSF are those with end-stage renal disease and patients undergoing dialysis. In such cases, the injection of a gadolinium-based contrast agent is only considered after a risk-benefit analysis has been completed, an alternate linear or macrocyclic agent issued and the dose limited to 0,1 mmol Gd/kg. Recently, recommendations from US and European agencies have converged. Learning objectives: to be familiar with the risk factors of CIN with iodinated contrast agents; to be familiar with hydration procedures for patients at risk of CIN; to be familiar with the diagnostic criteria of NSF; to be familiar with the classification of GBCA with regards to the risk of NSF; to be familiar with the contraindications of the different groups of GBCA.