具有广谱中和活性的HIV感染者血浆病毒膜蛋白基因及中和特征分析

目的 分析一例具有广谱中和活性的慢性HIV感染者病毒膜蛋白（Env）基因序列特征及中和特征。方法 使用单拷贝基因组扩增（SGA）方法从患者两个随访时间点血浆样本扩增env基因,测序后进行序列比对和进化树构建,分析env基因特征;将env基因克隆至pcDNA^TM3.1载体上,env质粒和骨架质粒pSG3△env共转染293T细胞制备假病毒,使用假病毒分别和两个时间点的自体血浆及6种不同的广谱中和抗体进行中和试验,分析Env蛋白的中和特征。结果 从两个时间点血浆样本中共获得50个全长env基因,进化分析显示两个时间点序列各自聚集成簇,第1个时间点膜蛋白序列有较短的V1区和较少的V1区糖基化位点数目,第2个时间点膜蛋白序列有更高的基因多样性。自体血浆不能中和同时期的假病毒,但能中和前期的假病毒;两个时间点的假病毒对PGT121、PGT135、2G12和10E8抗体均高度敏感,但对VRC01及12A21抗体呈现中和抗性。结论 该具有广谱中和活性的慢性HIV感染者病毒膜蛋白基因持续进化;感染者体内存在CD4bs类中和抗体的免疫压力。     

肠道嗜性和神经嗜性SIV毒株Gp120序列变异特点的比对分析

目的研究SIVmac239在不同中国恒河猴体内由于免疫压力出现的病毒Env区的变异进化情况。比对分析可形成AIDS相关肠病和脑病的SIV病毒其Gp120序列的差异及特点。方法四株SIVmac239感染晚期发病恒河猴PBMC中分离的病毒及两株神经嗜性SIVmac251病毒,通过有限稀释分离培养病毒单克隆,提取病毒RNA后反转录,PCR扩增病毒gp120序列进行系统进化树分析。同时分析两组不同嗜性毒株Gp120氨基酸序列及糖基化位点的变化情况。结果 SIVmac239在四只恒河猴体内发生不同的变异进化。肠道嗜性毒株与神经嗜性毒株氨基酸序列的差异集中在V1和V4区,肠道嗜性毒株在V4区与一个糖基化位点的增加,而神经嗜性毒株的糖基化位点的变化都发生在保守区C1、C2和C3。结论 SIV肠道嗜性和神经嗜性的增强分别与包膜蛋白Gp120上V4区糖基化位点的增加和C1区的糖基化位点缺失相关,两种嗜性的差异并不表现在与嗜性形成有紧密联系的V3区上。     

HIV-1功能性膜蛋白基因gp160的快速筛选和鉴定

目的 应用假病毒感染实验快速筛选可用于HIV-1中和抗体测定的功能性膜蛋白基因.方法 从HIV感染样品中扩增全长膜蛋白基因gp160,通过系统进化树分析确定其基因亚型,将HIV-1gp160阳性克隆与pSG3△Env质粒共转染293T细胞制备假病毒并对其感染能力进行分析,筛选功能性膜蛋白基因.结果 克隆和鉴定了我国HIV-1流行毒株的9个功能性膜蛋白基因,系统进化树分析表明其中包括4个CRF07_BC、2个CRF01_AE和3个B亚型毒株,这些gp160膜蛋白基因所制备的假病毒具有良好的感染性.结论 不同亚型HIV-1功能性膜蛋白gp160克隆为选择HIV-1中和抗体测定毒株提供了有价值的实验材料.     

HIV-1B’亚型及CRF07_BC重组毒株感染者中和反应特征分析

目的探讨我国主要流行的HIV～1B’和CRF07-BC重组毒株感染者中和反应特征。方法采用基于Env假病毒的以表达CD4及CCR5／CXCR4的TZM-bl细胞系为靶细胞的中和抗体测定方法,测定40例感染时间在3年左右的HIV—CRF07-BC重组毒株感染者及31例感染时间在10年以上的HIV-1B’亚型毒株感染者血浆对11株包括B（4株）、CRF07BC（3株）、CRF01-AE（3株）及中和敏感毒株（1株）Env假病毒的中和反应,并分析感染者血浆的中和宽度和中和强度。结果B’亚型毒株感染者在中和宽度及亚型交叉中和强度方面均高于感染时间相对较短的CRF07BC重组毒株感染者,B’亚型感染者对CRF01-AE亚型病毒的中和活性显著高于CRF07-BC重组型感染者：而CRF07-BC重组型感染者对同亚型病毒的中和强度则显著高于B’亚型感染者。结论HIV-1感染者中和抗体宽度以及对异源病毒的中和活性与感染时间正相关,适度的病毒抗原刺激有助于广谱中和活性的产生。     

一组分离自男男性行为人群HIV-1毒株的近似全长序列分析

目的对5名北京市男男性行为者体内人类免疫缺陷病毒-1（HIV-1）全基因组特征进行全面分析。方法采用淋巴细胞共培养法分离出病毒,并采用RT-PCR法扩增出其近似全长基因组,测序后对其基因序列特征进行深入分析。结果经系统进化和重组分析,分离出的5株病毒分属CRF07_BC、CRF01_AE和B亚型。病毒虽然分属不同亚型,但膜区糖基化位点的数目和分布位置都趋于保守,V3环顶端序列存在GPGR和GWGR两种形式,嗜性预测皆为M嗜性。结论男男性行为人群中HIV-1流行毒株存在CRF01_AE、CRF07_BC和B亚型。本研究获得的全长序列将为研究HIV-1各亚型在男男性行为人群中的传播和进化提供理论依据。     

HIV相关神经认知紊乱患者配对组织来源的HIV-1 env基因特征分析

目的：探讨人类免疫缺陷病毒相关神经认知紊乱（HIV associated neurocognitive disorder, HAND）患者不同组织中HIV-1 env基因的变异进化特征。方法通过国际HIV痴呆量表筛选出HAND患者,获得其血浆及脑脊液来源的HIV-1 env基因核苷酸序列,回顾性分析遗传多样性、氨基酸长度、潜在糖基化位点（PNGS）、辅助受体利用以及特征性氨基酸的特点及组织间差异。结果本研究纳入4例HAND患者共57条HIV-1 env基因V3-C4序列（血浆来源29条,脑脊液来源28条）。脑脊液中的HIV-1遗传多样性高于血浆来源毒株,差异有统计学意义（遗传多样性变异度中位数分别为0.053和0.007,P=0.043）。3例患者脑脊液与血浆来源的毒株均使用CCR5辅助受体,其中2例患者脑脊液（患者D 158 bp,患者F 158 bp）中的HIV-1 V3-C4区氨基酸长度中位数明显长于血浆来源（患者D 156 bp,P=0.017;患者F 157 bp,P=0.003）。脑脊液与血浆中HIV-1 env基因序列相比共有16个氨基酸位点存在明显差异,62.5%（10/16）分布在V4区。结论 HAND患者脑脊液和血浆中的HIV-1 env基因特征存在差异,脑脊液中的HIV-1准种遗传多样性更高。     

人类免疫缺陷病毒1型AE重组亚型毒株全长gp120基因序列分析

目的研究人类免疫缺陷病毒1型(HIV-1)AE重组型(CRF01-AE)毒株全长gp120基因的变异特征,为针对CRF01-AE重组型的免疫学研究和疫苗的研制提供帮助。方法应用巢式PCR对福建省2004至2005年期间100例HIV-1CRF01-AE感染者随机抽取21份血样的外膜蛋白全长gp120进行扩增、回收、克隆至T载体后测序,应用MEGA、DNASIS、CLUSTAL1·83、N-GLYCOSITE等软件对HIV-1全长gp120序列进行分析。结果基因系统树显示这21份样本属于CRF01-AE重组型HIV-1株,样本组内基因距离为9·5%±2·5%。V3环顶端四肽存在4种类型:GPGQ(71·43%),GPGR(19·05%),GPGH(4·76%),GQGQ(4·76%)。根据V3环关键氨基酸推测辅助受体使用情况的结果显示,16份(76·19%)可能使用CCR5作为辅助受体,1份(4·76%)样本可能使用CCR5/CXCR4序列,4份样本(19·05%)不能做出预测。21条HIV-1全长gp120氨基酸序列中,C1-C5比较保守,而V1-V5变化较大,其中V3相对保守。N糖基化位点分析发现大多数位点较为保守,少数发生糖基化位点丢失和增加。结论福建省大部分CRF01-AE重组型毒株与东南亚代表株关系密切,来源较为复杂,大部分CRF01-AE重组毒株为NSI型,gp120基因变异大,该研究为以后的病毒免疫学和疫苗的研究提供丰富的材料和基础数据。     

广东省人类免疫缺陷病毒1型CRF01_AE毒株膜区V3环氨基酸序列多态性分析

目的 分析广东省人类免疫缺陷病毒1型(HIV-1)CRF01_AE毒株膜蛋白第三高变区(V3环)氨基酸序列的特点。方法 提取HIV-CRF01_AE亚型感染者PBMC中前病毒DNA,巢式PCR扩增HIV-1膜基因c2v3c3区域后测序,对V3环氨基酸序列特征进行分析。结果 73份样本与HIV-CRF01_AE亚型代表毒株聚集在一起、与其他亚型标准毒株分开,进一步证实本研究73例HIV/AIDS患者感染的HIV-1毒株为CRF01_AE亚型。除6个位点氨基酸一致外,73例病人所有V3环序列的其他29个位点分别有2~8个不同的氨基酸出现;GPGQ为V3环顶端四肽的主要形式,占47.9%,其次为GPGK,占15.1%,四肽中未出现多态性的只有第三位氨基酸;预测HIV-1辅助受体为CCR5的有35例,占47.9%,其次为CXCR4的有34例,占46.6%,两者之间差异无统计学意义（P>0.05）。性传播是广东省CRF01_AE亚型HIV-1感染者的主要感染途径,占54.8%,其次是静脉吸毒,占30.2%。通过性传播途径感染的HIV-1毒株的辅助受体主要为CXCR4,占52.5%,辅助受体为CCR5的占40.0%,两者相比差异无统计学意义（P>0.05）。结论 广东省人类免疫缺陷病毒1型CRF01_AE毒株膜区V3环氨基酸存在较高的多态性,使用CCR5及CXCR4为辅助受体的HIV-1毒株比例相近。     

HIV-1广谱中和活性感染者假病毒的中和表型分析

目的分析HIV-1广谱中和活性感染者基于膜蛋白基因(env)的假病毒对自体血浆和代表性广谱单克隆中和抗体(bm NAbs)的中和表型。方法单基因组扩增(SGA)广谱中和活性感染者第0月血浆的env基因并克隆至pc D-NA~(TM)3.1 Directional TOPO表达载体,将env表达质粒与HIV-1骨架质粒p SG3△env共转染293T/17细胞制备假病毒,用假病毒分别与自体连续时间点血浆和代表性bm NAbs进行中和实验分析中和表型。结果共获得11株功能性假病毒,假病毒对8个连续时间点自体血浆的中和敏感性呈现先升高(第0~15月),后下降(第16~32月),之后再上升(第33~45月)的波动。所有假病毒对10E8、PGT121和VRC01中和敏感(IC_(50)6μg/m L);各有18.2%(2株)的假病毒对12A21和2G12高度敏感(IC_(50)1μg/m L);所有病毒均对PGT135呈中和抗性。结论假病毒对当前时间点血浆的中和敏感性低,对之后时间点血浆中和敏感性增强,提示感染者体内病毒与中和抗体的动态进化过程。同一时间点假病毒对特定bm-NAbs的敏感性差异明显,提示感染者准种毒株间中和表型的复杂性。     

无偿献血人群中HIV-1感染者env基因序列特征研究

目的 分析我国无偿献血人群艾滋病病毒I型（HIV-1）感染者病毒亚型及env基因C2-V4区序列特征。方法 针对我国13个省市献血人群筛选的115例HIV-1阳性标本,扩增HIV-1的env基因C2-V4区片段并测序;构建最大似然树（Maximum Likelihood）分析不同亚型env基因序列的进化关系;分析该基因区序列变异以及2G12、PGT135、PGT128中和表位特征。结果 从115例HIV阳性标本中成功扩增C2-V4区76份,经测序分析发现HIV-1基因亚型7种,其中,亚型CRF01_AE和CRF07_BC所占比例最高,分别占46.0%（35/76）和39.5%（30/76）,其他亚型CRF08_BC、B、CRF65_CPX、CRF59_01B、CRF61_BC所占比例分别为5.3%（4/76）、5.3%（4/76）、1.3% （1/76）、1.3% （1/76）、1.3% （1/76）。V3环顶端四肽存在GPGQ、GPGR、GPGK、GQGR四种类型,辅助受体以CCR5（90.8%）为主。97.1%的CRF01_AE毒株分别缺失2G12和PGT135抗体识别的相关糖基化位点,而CRF07_BC毒株100%缺失2G12抗体识别相关的糖基化位点,80%毒株未缺失PGT135抗体识别相关的糖基化位点。结论 我国献血人群HIV-1感染者以CRF01_AE和CRF07_BC为主要流行毒株;各毒株亚型V3顶端四肽以GPGQ变异型为主,且主要使用CCR5为辅助受体;2G12、PGT135及PGT128中和抗体表位呈现不同程度的变异。该研究为进一步开展针对献血人群的艾滋病预防与控制提供了参考数据。     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

Dr.Zhang Ren''''s Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

Luo Lingjie''''s Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.