抗AFP抗体—蓖麻毒素A链偶联物对靶细胞AH66的体外杀伤效应

本文应用流式细胞仪(FACStar~(PIns))的荧光分析技术,去检测与比较羊抗大鼠甲胎蛋白抗体(αAFP)与蓖麻毒蛋白A链(RTA)偶联物(αAFP-RTA)、RTA及αAFP和BTA的混和物(αAFP+RTA)对大鼠腹水型肝癌细胞AH66的体外杀伤作用。其αAFP-RTA、RTA和αAFP+RTA的剂量均以RTA的克分子浓度计算,致死50％靶细胞的浓度(IC_(50))分别为6×10~(-9.6)mol/L、6×10~(-8.4)mol/L、6×10~(-8.2)mol/L。αAFP-RTA与RTA的IC_(50)之比为1:12,αAFP-RTA与αAFP+RTA的IC_(50)之比为1∶14。相当于αAFP-RTA中抗体量的αAFP对AH66细胞不显示杀伤作用。实验证明,αAFP-RTA有明显的导向杀伤靶细胞的效应。上述实验结果与以台盼蓝除外试验所获得的结果基本一致。     

抗人B细胞MAb-737与蓖麻毒蛋白偶联物对肿瘤细胞选择性杀伤

具有针对B细胞类Ia抗原特异反应性MAb-737与蓖麻毒蛋白通过二硫键连接成免疫毒素ricin-737。体外0.1M乳糖存在时,此免疫毒素对含B细胞抗原的Burkitt氏淋巴瘤细胞系Raji细胞表现出选择性地强力杀伤,半数杀伤浓度(IC_(50))为5×10~(-11)M。同样条件,对不含靶抗原的对照细胞K_(562)没有显著杀伤,IC_(50)为10~(-3)M,毒性比前者低500倍。游离毒蛋白IC_(50)为5×10~(-3)M,比免疫毒素毒性低1000倍。动力学研究表明,免疫毒素作用1小时就可抑制靶细胞的增殖,并随处理时间延长,杀伤更趋显著。结果提示,免疫毒素ricin-737具有强烈地选择性杀伤靶细胞能力,可成为肿瘤治疗的候选药物。     

天花粉毒蛋白免疫毒素对黑色素瘤细胞的体外抑制

用单链致核糖体失活蛋白天花粉蛋白(TCS)和抗人黑色素瘤单抗(Ng76)构建了免疫毒素(Ng76—TCS),它对体外培养的人黑色素瘤细胞M21具有强烈的抑制活性,IC_(50)为5.6×10~(-10)mol/L,其毒性比游离的TCS 和Ng76的混合物高2000倍,比TCS 与正常小鼠免疫球蛋白的复合物(NIgG—TCS)高160倍,而它对非靶细胞HeLa 的IC_(50)为7.0×10~(-?)mol/L,毒性较靶细胞低125倍。结果表明,免疫毒素TCS—Ng76对体外培养的黑色素瘤细胞有很强的选择性杀伤作用.     

OX_(7-)肥皂草素免疫毒素的细胞毒性质研究

单链免疫毒素由於没有能与细胞膜上某些糖基结合的B链。所以非特异性毒性小,作为导向药物有一定的优点。肥皂草素是单链毒素中毒性较强的一种。本文比较了肥皂草素主要成分SO6与另一个成分SL连接到抗小鼠T细胞单抗OX_7上组成的免疫毒素的细胞毒性质。在无细胞体系中SL抑制蛋白质合成的活性较SO6高10倍,与OX_7抗体连接后的免疫毒素对靶细胞均有特异性毒性,但二种免疫毒素对靶细胞SL-2的IC_(50)都为3×10~(-10)M。动力学研究表明OX_7-SL对靶细胞的作用较OX_7·SO6快,说明前者杀伤靶细胞的效率较高。     

单克隆抗体HI_(30)萞麻毒素及其A链连接物的研究

用硫酸铵沉淀配合Sepharose 4 B柱亲和层析法从蓖麻子中提取到纯蓖麻毒素。该毒素 对小鼠LD_(50)为0.32μg。体外培养对CEM及L_(1210)细胞的IC_(50)分别为1.73×10~(-12)M和5.65×10~(-13)M。半乳糖能明显减弱毒素的细胞毒性。通过硫醚链连接制备的抗人T淋巴细胞表面分化抗原单抗 HI_(30)-完整毒素连接物基本保持了单抗与靶细胞CEM的结合能力并     

肝癌单抗—蓖麻毒素A链结合物的制备及其细胞毒性质

本文应用异型双功能基化合物SPDP与单克隆抗体反应,产生单克隆抗体-PDP,然后再与暴露巯基的RTA结合,制备肝癌单抗-RTA结合物。ELISA及毒素重组试验鉴定表明:结合物在10~(-9)mol/L浓度时与靶细胞仍有较好的结合能力,结合物中RTA保持良好活力。体外细胞毒结果表明:结合物对靶细胞具有良好的导向杀伤作用,在10~(-9)mol/L浓度时对人肝癌细胞株BEL7404的杀伤率为41.2％,对正常细胞SL_7的杀伤力很弱。     

抗CEA单抗-天花粉蛋白偶联物的研制及活性鉴定

应用SPDP化学偶联法,将天花粉蛋白通过二硫键交联与抗CEA单抗制备成免疫毒素,经Sepbadex G-150凝胶过滤和Blue Sepharose-4B离子梯度交换层析纯化,电泳鉴定显示这一免疫毒素纯度高,偶联蛋白克分子比合理。应用间接免疫荧光染色技术和ELISA法测定结果证明,其与CEA人结肠癌细胞(LoVo)具有特异结合反应,仍保留大部份原抗体活性。具有较好的导向性。体外毒性试验结果表明,免疫毒素对靶细胞(LoVo)有选择性的杀伤作用,50％细胞毒剂量(I.C50)为10~(-9)范围,比单用天花粉蛋白和天花粉蛋白与正常小鼠IgG-偶联物对靶细胞杀伤能力分别增强约150倍和2000倍,而对实验对照细胞(Hela)杀伤无增强作用。     

一种分离、培养扩增小鼠NK细胞方法的建立

目的:建立小鼠NK细胞的分离及培养扩增的方法。方法:用两步黏附分离法和磁珠活化的细胞分选(MACS)法,从小鼠脾脏的单个核细胞(MNC)中分离NK细胞,计数所得细胞的总数,并用FITC抗小鼠CD3和PE抗小鼠NK1.1单克隆荧光抗体染色后,用流式细胞仪(FACS)检测NK细胞的纯度。以YAC -1为靶细胞,用MTT比色法检测NK细胞的杀伤活性。结果:将两步黏附分离法分离的2×107个脾MNC培养5、10、15、20d,计数细胞的总数并检测CD3-NK1.1 细胞百分率,分别为0.5×107、1.4×107、2.6×107、3.0×107和18.36%、43.44%、55.68%、60.03%。效靶细胞25∶1时,NK细胞的杀伤率分别为54.38%、66.54%、79.38%和83.86%,明显高于脾MNC(41.93%)(P<0.01)。MACS法纯化前后细胞的总数和CD3-NK1.1 细胞的百分率,分别为1.0×108、1.5×106和2.54%、93.60%;但纯化后的细胞在体外培养20d时,只扩增到1.9×106个。结论:用两步黏附分离法分离的小鼠NK细胞,培养2～3wk可获得大量的NK细胞,纯度可达55%～60%;在效靶细胞为25∶1时,NK细胞对YAC1细胞的杀伤率约为80%。     

抗T细胞免疫毒素的T细胞清除效率及其对造血细胞的影响

抗T细胞免疫毒素主要用于同种异体骨髓移植中清除T细胞,减轻移植物抗宿主反应(GVHD),提高移植成功率,也可用于白血病自体骨髓移植中清除残留白血病细胞,减少移植后白血病复发的可能性。本实验结果表明抗T细胞免疫毒素在10~(-8)M浓度对靶细胞的杀伤达3.0log以上,即可清除99.99%的靶细胞,并可有效地抑制周血T细胞转化功能和骨髓T细胞分泌IL—2的功能,同时对骨髓粒单系和红系造血细胞的增殖无明显的抑制作用。     

胃癌单抗-RTA结合物的制备及其细胞毒特性研究

本文报道鼠抗人胃癌单克隆抗体-蓖麻毒素A链结合物的制备及其对肿瘤靶细胞的杀伤作用。抗体与3-(2-吡啶二巯基)丙酸-N-琥珀酰亚胺酯(SPDP)反应引入2-吡啶二硫化物,然后与巯基暴露的A链反应,得到抗体-A链结合物。ELISA结果表明,抗体与毒素A链连接是成功的,结合物中抗体保存较好的与靶细胞结合活性。结合物对靶细胞具有较强的选择性杀伤作用,在1×10~(-9)M水平,对人胃癌细胞KATO Ⅲ蛋白质合成抑制率为71％,而对非靶细胞作用很弱。结果表明,该单抗具有较好毒素A链导向作用。     

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