Ⅰ型人类免疫缺陷病毒包膜gp120与乙型肝炎病毒表面抗原免疫原性比较

目的 比较Ⅰ型人类免疫缺陷病毒(HIV-1)及乙型肝炎病毒(HBV)包膜蛋白初次免疫及加强免疫后诱导产生抗体的规律,为提高HIV-1包膜蛋白诱导保护性抗体产生能力提供创新思路.方法 以10周龄雌性C57BL/6小鼠为动物模型,分别用HIV-1 06044毒株gp120三聚体(gp120T)、HBV表面抗原(HBsAg)蛋白与AddaVax佐剂免疫小鼠,背部皮下注射,共免疫3次,每次免疫间隔3周,第一、第二次免疫后7d和第三次免疫后3d、7d取血;第一次免疫后7d、第三次免疫后3d、7d取脾组织.用酶联免疫吸附实验(ELISA)及酶联免疫斑点实验(ELISpot)方法检测免疫小鼠血浆特异性结合抗体滴度及抗体分泌细胞(ASC)数量.结果 gp120T和HBsAg两种蛋白初次免疫后,动物均未产生明显的特异性抗体.两种蛋白加强免疫后特异性抗体水平明显升高,gp120T一次加强免疫及两次加强特异性抗体滴度逐渐升高,而HBsAg一次加强抗体滴度已经接近两次加强的水平.二次加强免疫后,gp120T和HBsAg免疫鼠脾脏特异性ASC数量差异不显著.结论 HIV-1包膜gp120T加强免疫诱导抗体水平达到高峰慢于HBsAg加强免疫,即加强免疫后gp120T诱导的回忆反应慢于HBsAg.     

原代HIV-1包膜糖蛋白修饰体诱导小鼠产生高滴度gp120抗体

目的 探索1型人类免疫缺陷病毒(HIV-1)包膜糖蛋白修饰对包膜免疫原性的的影响.方法 通过PCR扩增获得原代HIV-1 06044株包膜gp120基因及其突变体gp120/W427S基因,并构建gp120三聚体蛋白真核表达载体pcT-gp120和pcT-gp120/W427S,重组表达载体体外瞬时转染人胚肾HEK2...     

HIV-1流行株去糖基化修饰gp41突变体的构建及其表达和纯化

目的:构建HIV-1ⅢB标准株、中国大陆地区主要流行株CRF-BC和海南省流行株CRF-AE野生型和突变型gp41表达载体,获得gp41重组蛋白。方法:利用基因定点突变的方法,将HIV-1ⅢB、CRF-BC和CRF-AE流行株野生型gp41基因中特定的N611Q糖基化位点天冬酰胺Asn突变为谷氨酰胺Gln,使该位点去糖基化。利用基因工程技术构建野生型pc DNA3. 1-gp41和突变型pc DNA3. 1-Mgp41表达载体。取测序正确的重组质粒转染293 T细胞,重组蛋白经Ni-NTA亲和层析柱纯化后采用免疫杂交方法进行分析。结果:测序结果显示野生型gp41的N611糖基化位点均由天冬酰胺(Asn)的编码基因(AAU)突变为谷氨酰胺(Gln)的编码基因(GAA),成功构建了野生型和突变型gp41表达载体。琼脂糖凝胶电泳分析显示野生型和突变型HIV-1 gp41基因大小约为700 bp,Western blot免疫杂交结果显示在26 k D处出现目的条带,与预期一致。结论:成功构建了HIV-1流行株去糖基化修饰gp41突变体,为研究去糖基化修饰gp41蛋白与疾病发生发展的分子机制奠定基础。     

含密码子优化型HIV-1 gp120基因重组腺病毒的构建及其免疫效果研究

目的 构建能表达野生型和密码子优化型人免疫缺陷病毒Ⅰ型(HIV-1)B亚型中国流行株gp120基因的非复制型腺病毒。方法 按哺乳动物细胞偏好的密码子对HIV-1B亚型中国流行株Ch gp42的gp120基因进行优化，合成优化基因。将野生型和密码子优化的gp120基因插入穿梭质粒，再与腺病毒骨架质粒pAdEasy-1共转化E．coli BJ5183，获得重组子，转染293细胞后获得重组病毒。分别以两种重组腺病毒疫苗免疫小鼠，ELISA检测小鼠血清中的特异性抗体，乳酸脱氢酶法检测小鼠细胞毒性T淋巴细胞(CTL)反应。结果 获得两株重组腺病毒rAd-wt．gp120和rAd．mod．gp120，能正确表达Gp120。rAd-mod．gp120比rAd-wt．gp120蛋白表达水平明显提高。重组腺病毒免疫小鼠后能产生HIV-1特异性的抗体及CTL反应，rAd-mod．gp120组明显优于rAd-wt．gp120组。结论 成功构建了表达野生型和密码子优化的HIV-1 gp120基因的重组腺病毒，能诱导HIV-1特异性体液和细胞免疫反应。     

HIV-1包膜蛋白V2区突变对V3区特异的中和抗体的影响

目的 研究HIV-1包膜蛋白V2区突变对V3区特异中和抗体与HIV-1中和作用的影响.方法 使用假病毒中和试验测定L175P突变对于V3区中和抗体对HIV-1中和能力的影响,ELISA方法测定上述中和抗体对gp120蛋白单体的结合力.结果 V2区突变L175P可以改变多个V3区特异性中和抗体对病毒的中和能力,但这些中和抗体对gp120蛋白单体的结合力不因V2区突变而改变.结论 V2区的L175P突变改变了gp120蛋白天然三聚体的结构,促进了V3区抗体的中和能力.

Abstract：

Objective To study the impact of V2 mutations on neutralizing ability of HIV-1-specific neutralizing antibodies. Methods We tested the influence of L175P mutation to the neutralizing ability of V3-specific antibodies by pseudotype virus and the binding affinity of those V3-spesific antibodies to gpl20 monomer by ELISA. Results We found L175P mutation changed the neutralizing ability of V3-specific antibodies. However, L175P mutation showed no effects on the binding affinity of these antibodies to gpl20 monomer. Conclusion Our results revealed the L175P mutation at V2 loop changed the natural trimmer structure of gp120 and enhanced the neutralizing ability of V3-specific antibodies.     

人类免疫缺陷病毒1型(HIV-1)糖蛋白gp120部分糖基化位点突变对其抗原性影响

目的 研究HIV-1包膜糖蛋白gp120特定糖基化位点突变后对gp120抗原性的影响,并推测糖基化位点突变对结构的影响.方法 采用连接PCR方法,将获得的野生型gp120中选定N糖基化位点的Asn定点突变为Cln,使该位点去糖基化,构建DNA疫苗,免疫小鼠,ELISA检测鼠血清中的抗体,进行统计学分析,推测糖基化位点突变对gp120抗原性和结构的影响.结果 2组糖基化位点突变的gp120诱发非V3特异性抗体的能力较之野生型gp120有明显提高,但诱发V3特异性抗体能力没有显著提高,7个位点突变的gp120诱发V3特异性抗体能力有很大降低.结论 gp120部分糖基化位点的突变在一定程度上能提高或改变gp120的抗原性,这可能由突变导致的构象变化和/或糖基化寡糖链移除使原来被遮盖的抗原表位暴露引起.     

HIV-1广谱中和性单克隆抗体的研究进展

从HIV感染患者血清中分离的针对HIV-1包膜上保守表位的中和性单克隆抗体(mAb)很多,识别的表位分别是HIV-1gp120上CD4结合位点(CD4bs),gp41的跨膜前域MPER,gp120表面包膜的高甘露聚糖,gp120上V2/V3环,这些抗体对HIV-1疫苗免疫原的研究设计至关重要.本文主要综述了各中和性单克隆抗体的结构特征,识别的表位,抗体与病毒包膜之间的相互作用.     

HIV-1中和抗体b12的研究进展

HIV-1由于其外膜糖蛋白结构多变可以很容易逃避抗体介导的中和作用.b12抗体是目前少数几种被证明具有广谱抗HIV-1活性的中和抗体,近来关于gp120结构研究表明在gp120上有一个保守的中和位点,且b12抗体可以和这个位点结合从而阻断靶细胞CD4与gp120作用,这一发现预示b12抗体对于新一代艾滋病疫苗设计具有重要的作用.本文即对b12抗体的有关研究进展作一综述.     

HIV-1中和抗体b12的研究进展

HIV-1由于其外膜糖蛋白结构多变可以很容易逃避抗体介导的中和作用.b12抗体是目前少数几种被证明具有广谱抗HIV-1活性的中和抗体,近来关于gp120结构研究表明在gp120上有一个保守的中和位点,且b12抗体可以和这个位点结合从而阻断靶细胞CD4与gp120作用,这一发现预示b12抗体对于新一代艾滋病疫苗设计具有重要的作用.本文即对b12抗体的有关研究进展作一综述.     

HIV-1中和抗体b12的研究进展

HIV-1由于其外膜糖蛋白结构多变可以很容易逃避抗体介导的中和作用.b12抗体是目前少数几种被证明具有广谱抗HIV-1活性的中和抗体,近来关于gp120结构研究表明在gp120上有一个保守的中和位点,且b12抗体可以和这个位点结合从而阻断靶细胞CD4与gp120作用,这一发现预示b12抗体对于新一代艾滋病疫苗设计具有重要的作用.本文即对b12抗体的有关研究进展作一综述.     

Level of functioning of siblings and parents of probands of varying degrees of retardation

A further analysis of already published data supports the position that retardates of low ability level less frequently have retarded siblings, retarded parents, and parents low in occupational level than do retardates higher in ability level. The analysis supports the position that there are two types of retarded individuals, persons retarded as a result of gene or chromosomal anomalies, brain injury, etc., who more frequently occur in the lower-level retardate group, and persons whose retardation represents polygenic segregation, who more frequently occur in the higher-level group.     

局部注射辛伐他汀对骨质疏松大鼠股骨髁骨小梁的改建效应

背景：局部注射具有成骨作用的辛伐他汀,可显著增加骨质疏松大鼠股骨颈及股骨髁部的骨密度及力学强度,分析局部注射辛伐他汀对股骨髁骨小梁的影响。 目的：进一步研究骨质疏松大鼠股骨内局部注射辛伐他汀对股骨髁骨小梁的影响。为将辛伐他汀应用于临床骨质疏松局部治疗提供实验基础。 方法：18只雌性SD大鼠双侧卵巢切除后3个月,制备大鼠骨质疏松模型。实验大鼠随机数字表法均分为3组,分别在实验大鼠的右侧股骨髓腔内单次注射辛伐他汀溶液5 mg、10 mg,对照组单纯注射空白载体。分别在注射后1个月处死大鼠并取材。Micro-CT扫描并定量分析骨组织形态变化。 结果与结论：给药后1个月,Micro-CT扫描结果显示,辛伐他汀治疗组的骨微结构参数如骨皮质厚度、骨小梁密度及连接率明显优于对照组。说明疏松骨骼单次注射小剂量辛伐他汀可显著促进股骨髁部骨小梁改建,改善骨骼微结构,可为强化局部、防治骨质疏松骨折的新选择进一步提供实验基础。     

石斛属民族药用植物的分类及生药学研究

中药石斛早在《神农本草经》中就被列为上品,其应用历史悠久,具有养阴生津、补肾益气、润喉护嗓、活血明目、抗癌防老等功效。本文对国内外石斛属民族药用植物的分类及生药学研究进行了综述。     

膝关节骨性关节炎的关节镜治疗体会

目的我科自2001～2005年4月对59例67膝的骨性关节炎（0A），进行关节镜检查及镜下清理术。方法镜检：膝关节骨性关节炎伴不同程度滑膜炎，滑膜皱壁粘连纤维片，关节软骨不同程度破坏。镜下清除增生滑膜，松解粘连，去除剥脱软骨、修复关节面。结果随访2月～4年，平均1.5年，术后综合评估2月～1年，优良率94％；1～2年，优良率79.2％；2年以上优良率54.8％。结论关节镜对膝关节骨性关节炎诊断能提供了比较全面的资料，并对骨性关节炎早期有良好疗效，具有创伤小、恢复快、并发症少和重复治疗等优点。     

Anatomical study of distribution of valves of the cutaneous veins of adult''s limbs

Since the cutaneous veins of the four limbs have been used as autogenous grafts in the treatment of cardiovascular disorders, knowledge of distribution of the valves is increasingly required depending upon the use. In the gross anatomical study of distribution of valves of the trunci venae, there has been argument about locational relationships between the venous roots and the valves in the vicinity of the roots and the inter-valvular distance. However, there have been only few reports discussing detailed information about valves of the cutaneous veins of the four limbs. The authors observed patterns of distribution of the cutaneous venous valves of the four limbs of cadavers prepared for practice in anatomy. The following parts were excised from each cadaver: the cephalic, basilic, and the great saphenous veins, which originate from the acral venous network and flow into the proximal deep veins, and the venous roots communicating with these veins. An incision was made on each excised vein in the direction of the long axis under observation with a stereoscopic microscope, and the inter-valvular distance and the distance between the valve and the orifice of venous root in the vicinity of the valve were measured. The inter-valvular distance varied with type of the truncus venae, and it varied according to area even in the same truncus venae.     

Pathways of the realization of immunomodulating effects of neurotransmitters

Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 115, N ^o 4, pp. 401–404, April, 1993     

Characterization of sterols of three digenetic trematodes of buffalo

Sterols of three digenetic trematodes were isolated and characterized by infrared and 1H nuclear magnetic resonance spectroscopies, and gas-liquid chromatography and gas chromatography-mass spectrometry. Sterols identified were cholesterol, cholestanol, 24-methylcholesterol, 24-methylcholestanol, 24-ethyl-22-dehydrocholesterol, 24-ethyl-22-dehydrocholestanol, 24-ethylcholesterol and 24-ethylcholestanol.     

Mechanisms of generation of slow components of bioelectric activity in diagnostics of functional state of the gastrointestinal tract

New electrophysiologic devices were used for the diagnostics of the state of excitable structures in the gastrointestinal tract and correction of their motor functions. Bioelectrical and biomechanical activities form the basis of functioning of internal organs. The mechanisms of generation of slow bioelectrical activity that are important for clinical and physiological studies are described. One of these mechanisms is a capacitance parametric transducer converting the energy of contractions into specific electric signals reflecting muscle functions. Another mechanism results from slow oscillations of resting potentials of interrelated excitable cells in large neuromuscular structures of internal organs. The elaborated procedure is efficient for preventing early postoperative paresis of the gastrointestinal tract. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 10, pp. 448–452, October, 1999     

胃小弯侧血管弓的MDCT解剖

目的：研究正常胃小弯侧血管弓及其属支血管的多排螺旋CT表现。方法：回顾性分析51例正常成人的上腹部CT资料,观察胃小弯侧血管弓及其属支血管的显示情况、血管弓形态及其与胃壁的关系。结果：51例均显示胃小弯侧血管弓,血管弓贲门部属支、胃体部属支和幽门部属支分别显示46例、15例和10例;血管弓与胃壁紧邻41例。结论：MDCT检查可良好显示正常胃小弯侧血管弓及其属支血管。     

Assessment of the efficacy of treatment of hemangiomas

Assessment of the aftereffects of cryoexposure and ultrahigh-frequency cryoexposure on hemangioma tissue of various types, cavernous and squamous, showed a higher cryogenic effect in hemangiomatous tissue preexposed to ultrahigh-frequency waves. A quantitative criterion is proposed for assessing the efficacy of the studied methods of exposure. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 6, pp. 669–672, June, 1995