重组人bFGF的原核表达及其高效价抗血清的制备

目的 以重组人碱性成纤维生长因子为免疫原，制备高效价抗hbFGF抗血清。方法通过PCR方法改造5’编码区的12个密码子，构建hbFGF’原核表达载体并在大肠杆菌(E．coli)中表达，以纯化的hbFGF、免疫新西兰兔，制备高效价抗血清，用于重组hbFGF、的免疫印迹分析。结果经过改造的hbFGF基因在E．coli中获得较高水平表达。从可溶性部分纯化得到纯度95％以上的重组hbFGF，以该重组蛋白免疫兔子，在二次加强后以间接ELISA检测抗血清效价可达1：512000。免疫印迹分析显示该抗血清与E．coli中表达的重组hbFGF、和标准hbFGF、均有特异性反应，但与某些细菌蛋白存在弱交叉反应，经E．coli菌体蛋白吸附的抗血清，与菌体蛋白的弱交叉反应消失。结论以纯化的重组hbFGF为免疫原制备了高效价的特异性抗血清，经菌体蛋白吸附可消除存在的交叉反应性。     

Cross-neutralization of the neurotoxicity of Crotalus durissus terrificus and Bothrops jararacussu venoms by antisera against crotoxin and phospholipase A2 from Crotalus durissus cascavella venom.

We have previously demonstrated that rabbit antisera raised against crotoxin from Crotalus durissus cascavella venom (cdc-crotoxin) and its PLA2 (cdc-PLA2) neutralized the neurotoxicity of this venom and its crotoxin. In this study, we examined the ability of these antisera to neutralize the neurotoxicity of Crotalus durissus terrificus and Bothrops jararacussu venoms and their major toxins, cdt-crotoxin and bothropstoxin-I (BthTX-I), respectively, in mouse isolated phrenic nerve-diaphragm preparations. Immunoblotting showed that antiserum to cdc-crotoxin recognized cdt-crotoxin and BthTX-I, while antiserum to cdc-PLA2 recognized cdt-PLA2 and BthTX-I. ELISA corroborated this cross-reactivity. Antiserum to cdc-crotoxin prevented the neuromuscular blockade caused by C. d. terrificus venom and its crotoxin at a venom/crotoxin:antiserum ratio of 1:3. Antiserum to cdc-PLA2 also neutralized the neuromuscular blockade caused by C. d. terrificus venom or its crotoxin at venom or toxin:antiserum ratios of 1:3 and 1:1, respectively. The neuromuscular blockade caused by B. jararacussu venom and BthTX-I was also neutralized by the antisera to cdc-crotoxin and cdc-PLA2 at a venom/toxin:antiserum ratio of 1:10 for both. Commercial equine antivenom raised against C. d. terrificus venom was effective in preventing the neuromuscular blockade typical of B. jararacussu venom (venom:antivenom ratio of 1:2), whereas for BthTX-I the ratio was 1:10. These results show that antiserum produced against PLA2, the major toxin in C. durissus cascavella venom, efficiently neutralized the neurotoxicity of C. d. terrificus and B. jararacussu venoms and their PLA2 toxins.     

四型念珠菌与抗念珠菌血清的交叉免疫反应

目的 分析4型不同念珠菌抗血清的免疫应答状况,为念珠菌疫苗的研制奠定实验基础.方法 将白色念珠菌、热带念珠菌、吉利蒙念珠菌、假热带念珠菌4型念珠菌分别超声破碎,0.5％甲醛溶液灭活,加入弗氏完全佐剂制成灭活全菌疫苗.分别在1、4、8、12周腹腔注射免疫小鼠,4周后收集小鼠抗体血清,分别按10、100、1000倍稀释,再分别加入含200μl4种念珠菌的沙氏培养液中,观察其抑菌作用.结果 所有抗体均为10倍效果最佳,1000倍基本无效.白色念珠菌抗体血清对白色念珠菌、吉利蒙念珠菌、热带念珠菌有明显的抑菌作用（P＜0.01）;热带念珠菌抗体血清对白色念珠菌和热带念珠菌有抑菌作用（P=0.01）;吉利蒙念珠菌抗体血清对吉利蒙念珠菌、热带念珠菌有抑制作用（P＜0.01）;假热带念珠菌抗体血清对热带念珠菌、假热带念珠菌有抑制作用（P＜0.01）.结论 小鼠念珠菌免疫抗体血清对念珠菌的生长有一定的抑制作用.     

Characterization of an antiserum to synaptic glomeruli from rat cerebellum

Rabbit anti-rat cerebellar synaptic glomeruli antiserum when absorbed with non-neural tissues reacts only with neural tissues when tested by indirect immunofluorescence on tissue sections. Further absorption with forebrain results in an antiserum which detectably reacts only with synaptic glomeruli and soma of Purkinje cells of both rat and mouse. The developmental expression of the synaptic glomeruli antigen(s) parallels the formation of synapses between mossy fibers and granule cells. Immature synaptic contacts do not contain recognizable antigent(s), whereas only at postnatal Day 15 glomeruli become antigen-positive. At this stage antigen in Purkinje cells is no longer carried in their dendrites, but becomes confined to the cell soma. Staggerer mutant mice still express the immature pattern of antigen distribution on postnatal Day 18.     

Inhibition by neuropeptide Y of fentanyl-induced muscular rigidity at the locus coeruleus in rats

The aim of this study was to evaluate the effects of centrally administered neuropeptide Y (NPY) on muscular rigidity induced by fentanyl in Sprague–Dawley rats. They were anesthetized with ketamine (120 mg/kg, i.p.) and their lungs were mechanically ventilated. Intravenous administration of fentanyl (100 μg/kg) consistently evoked a significant increase in the electromyographic activity (EMG) recorded from the sacrococcygeus dorsalis lateralis muscle. This implied muscular rigidity was appreciably attenuated by intracerebroventricular administration of NPY (4 nmol/2.5 μl). Microinjection of NPY (40 or 160 pmol/50 nl) into the bilateral locus coeruleus (LC) elicited an inhibition of the EMG activation induced by fentanyl in a dose-dependent manner. Microinjection of 160 pmol NPY plus antiserum against NPY (NPY_Ab, 1:20) into the LC failed to suppress fentanyl-induced muscular rigidity. However, this implied muscular rigidity was not affected by NPY plus normal rabbit serum (NRS, 1:20). In addition, NPY_Ab or NRS per se had no substantial effect on the rigidity. Microinjection of NPY (160 pmol) into the areas adjacent to the LC did not attenuate the rigidity. Our results suggest that the centrally administered NPY attenuated fentanyl-induced muscular rigidity by acting at the LC, but endogenous NPY may not be involved in this process.     

胰岛素酶联免疫测定法的研究快速制备高质量胰岛素抗血清(Ⅰ)

本文报告制备胰岛素抗血清的新方法。每只免疫动物的胰岛素用量为200～300μg.免疫时间60天。经放射免疫分析,10株抗血清的50%B/T稀释度均大于1:10万,最高者达1:26万。用计算机进行Scatchard分析,10株抗血清的Ka_1均大于1.5×10~(12)L/M,Ka_2均小于4×10~(?)L/M。[Ab~·]_1,在0.5～1.6×10~(-12)M/L之间,[Ab~°]_2均大于1×10~(-11)M/L。10株抗血清与前胰岛素、胰多肽和胰高血糖素进行放免分析,均无交叉反应。     

抗p30血清的制备与鉴定

作者用分离和纯化获得的p30免疫家兔,制备特异性抗p30血清。鉴定结果表明,该抗血清仅与p30发生反应,与猪、狗、牛、羊、兔、大鼠、小鼠、鸡等8种动物精液;猪、狗、牛、鸡等4种动物血清;人阴道分泌液、初乳、血清、唾液、尿液等5种人体液;人阴道粘膜、子宫内膜、蜕膜、绒毛、卵巢、乳腺、睾丸、附睾、脑、扁桃、甲状腺、胸腺、心、肺、脾、肝、胰、胃、小肠、结肠、肾上腺、肾与膀胱等23种人体组织均无交叉反应,具有高度的种属特异性和男性生殖器官特异性。     

Failure to detect radioimmunoassayable arginine vasotocin in mammalian pineals

The presence of arginine vasopressin (AVP) and arginine vasotocin (AVT) in pineal and neural lobes of a variety of species was evaluated by radioimmunoassay using two antisera. One antiserum recognizes only AVP, while the other cross-reacts 100% with both AVP and AVT. Only traces of AVP were detected in toad and chicken pituitaries and chicken pineals, tissues found to contain abundant AVT. On the other hand, pineals from rats and rabbits, while containing measurable quantities of AVP, did not contain detectable levels (less than 6 pg) of AVT. These results fail to demonstrate the presence of AVT in rat and rabbit pineals and strongly question the role of AVT as a physiological pineal hormone in mammals.     

抗Dystrophin特异性抗血清Anti 5－7的制备

应用作者自行制备的含ＤＭＤ基因中段缺失突变热点区ｃＤＮＡ片段的克隆菌株ＴＡＰ１０６（ｐＳＤ），表达出ＤＭＤｃＤＮＡ５－７蛋白，作为抗原制备特异性抗血清。利用彻底变性的、由ＳＤＳ－ＰＡＧＥ凝胶直接匀浆制备的抗原、和电洗脱纯化使处于相对天然状态的抗原分别免疫兔，得到各为１：２５６０和１：１２８０的高效价抗血清Ａｎｔｉ５－７，为我国ＤＭＤ／ＢＭＤ蛋白诊断创造了条件。