微囊化异种甲状旁腺组织移植的研究

目的探讨微囊化异种甲状旁腺(PTG)组织移植对Wistar大鼠甲状旁腺功能低下的治疗作用及微囊的通透性。方法 40只去甲状旁腺Wistar大鼠随机分为微囊组、非微囊组、空囊组、空白对照组。用海藻酸-钡交联微囊包裹兔甲状旁腺组织,移植至Wistar大鼠肾包囊。移植后每隔2周取血测血钙,移植后第16周取移植物进行透射电镜检查及T淋巴细胞、大鼠IgG抗体的免疫组织化学染色。结果微囊组移植后第4周血清钙由(1．62±0．04)mmol／L恢复至正常水平 (2．2～2．6)mmol／L,9例维持至观察期结束(P<0．01),非微囊组、空囊组及空白对照组血清钙差异无统计学意义(P>0．05)。第16周取出移植物检测显示移植物活性良好,微囊周围可见较多T 淋巴细胞浸润,囊壁及囊内IgG抗体染色阳性。结论海藻酸-钡交联微囊可对甲状旁腺组织起到有效的保护作用,使甲状旁腺组织较长时间存活并发挥正常功能。但海藻酸-钡交联微囊并未减轻受体免疫排斥反应的激活且未有效的免疫隔离IgG抗体。     

IL-4与IL-10联合诱导异种骨移植免疫耐受的体外研究

目的　探讨 IL - 4和 IL - 10在诱导异种骨移植免疫耐受中的作用。方法　反应细胞为 BAL B/c小鼠脾淋巴细胞 ,刺激细胞为新西兰白兔血淋巴细胞 ,刺激抗原为兔骨上清液。采用经典的混合淋巴细胞培养法及骨上清液与淋巴细胞混合培养法作为异种骨移植的体外实验模型。在各培养液中分别加入 IL - 4、IL - 10及两者联合应用 ,通过测定其 3H- Td R掺入率 ,观察不同细胞因子对刺激淋巴细胞增殖的影响。结果　无论在细胞刺激组还是骨上清液刺激组 ,IL- 4对淋巴细胞增殖均有显著抑制作用 (P<0 .0 0 1和 P<0 .0 5 ) ,IL- 10未表现出抑制作用 (P>0 .0 5 )。在两组 IL- 4和 IL - 10联合应用均产生比 IL - 4单独应用更为明显的细胞增殖抑制作用 (P<0 .0 0 1和 P<0 .0 5 )。结论　 IL - 4对由细胞或骨上清液刺激产生的淋巴细胞增殖均有很好的抑制作用 ,IL- 10没有表现出抑制作用 ;IL- 4与 IL- 10联合应用有协同抑制作用。     

中华眼镜蛇蛇毒对大鼠异种心脏移植超急性排斥反应的影响

试用中华眼镜蛇蛇毒消耗补体，观察其对豚鼠到ＳＤ大鼠异种心脏移植超急性排斥反应的影响。一次性给予小剂量ＣＣＶ腹腔注射后可明显降低大鼠血清补体溶血活性，４－２４小时后渐降至给药前１／２以下水平。ＣＣＶ可明显延长异种移植心脏４ 存活时间达２．５－３６小时，对照组仅为２５分钟；ＣＣＶ与脾切除，前列腺素Ｅ１联用可进一步延长移植心的存活时间，最长１例达４０小时。     

Heterologous Expression of Rat Testis GABAA Receptor β3t Splicing Variant in CHO Cells

Objective To characterize a possible retention function of unique sequence in the 5'end of rat testis GABAA receptor β3t splicing variantMethods Rat testis GABAA receptor β3t splicing variant cDNA was cloned and two eukaryotic expression recombinant plasmids of pEGFP-N1 and pEGFP-C1 were constructed respectively by fusing green fluorescent protein to the N or C-terminus of β3t isoform. The recombinant plasmids were transfected into CHO cells by calcium phosphate co-precipitation method. Fluorescence microscope and laser confocal microscope were used to analyze localization of β3t in the transfected cells. ConA-Texas-Red was used to label cell ER and the localization of rat testis β3t splicing variant in CHO cells was determined.Results When rat testis β3t splicing variant was expressed in CHO cells, two expression patterns were delineated, the distributions of uniform and mainly discrete intracellular compartments respectively. The chimera product failed to be translocated into the cell surface when expressed in CHO cells; whereas the β3 subunit of rat brain was incorporated into the plasma membrane.Conclusion The inability of β3t to target into the ER may be a consequence of the unique 25 specific amino acid segments in the N terminus.     

微囊化大鼠胰岛异种移植的实验研究

目的　通过动物实验明确微囊化胰岛是否具有免疫隔离作用。方法　SD大鼠胰腺原位消化 ,Ficoll间断密度梯度离心法纯化、分离胰岛 ,气流吹喷制作海藻酸钠 /聚赖氨酸 /海藻酸钠(APA)微囊化大鼠胰岛 ,比较微囊化与未微囊化胰岛的胰岛素释放试验 ;将微囊化 (实验组 )与未微囊化 (对照组 )大鼠胰岛植入链脲佐菌素 (STZ)诱导的I型糖尿病小鼠中 ,作两组间血糖正常持续时间比较。结果　实验组与对照组的胰岛素释放试验差异无显著性 (P >0 .0 5 ) ;实验组血糖正常持续时间为 2 3～ 6 5d(平均 48d) ,对照组为 3～ 6d(平均 5d) ,两组差异有极显著性 (P <0 .0 1)。已排斥的实验组小鼠腹腔灌洗发现部分微囊化胰岛存活 ,部分已坏死 ,但微囊膜皆完整 ,囊壁无纤维化。结论　微囊具有良好的免疫隔离作用 ,可使胰岛移植物存活时间明显延长。同时推测微囊内移植物死亡与细胞因子、自由基作用或营养不足等有关。     

Rat Cytomegalovirus Infection Interferes with Anti-CD4 mAb-(RIB 5/2) Mediated Tolerance and Induces Chronic Allograft Damage

In order to assess the role of heterologous immunity on tolerance induction (TI) by signal 1 modification, the influence of rat cytomegalovirus infection (RCMVI) on TI by a non-depleting monoclonal anti-CD4 mAb (monoclonal antibody) (RIB 5/2) in a rat kidney transplant (KTx) model was investigated. Orthotopic rat KTx (Dark Agouty (DA)-->Lewis (LEW)) was performed after TI with RIB 5/2 [10 mg/kg body weight (BW); day -1, 0, 1, 2, 3; i.p. (intraperitoneal route)]. RCMVI (5x10E5 Plaque forming units [PFU] i.p.) was simultaneously conducted to KTx, 50 days after KTx, and 14 days before and after KTx. RIB 5/2 induced robust allograft tolerance even across the high-responder strain barrier. RCMVI broke RIB 5/2-induced tolerance regardless of the time of RCMVI but did not induce acute graft failure during the 120 days follow-up. RCMVI induced a significant chronic deterioration of allograft function (p<0.01) and enhanced morphological signs of chronic allograft damage (p<0.05). Cellular infiltrates and major histo-compatibility complex (MHC)-expression were more pronounced (p<0.05) in the infected groups. RCMVI induced not only RCMV-specific T-cell response but also enhanced the frequency of alloreactive T cells. RCMV interferes with anti-CD4 mAb-induced tolerance and leads to chronic allograft damage. The data we presented suggest a potentially important role of viral infections and their prophylaxis in clinical TI protocols.     

酒精胎骨混悬液注射移植促进骨愈合实验研究

用酒精兔胎骨混悬液为植骨材料，以兔双侧桡骨中段３ｍｍ宽骨缺损横断骨折及肱三头肌为动物模型，通过注射将骨植入实验侧骨折端及肌肉内，对侧注入等量生理盐水作自身对照。术后进行免疫学、放射学、组织学及生物力学检查，结果表明：植骨不引起明显免疫排斥反应；植骨侧新骨形成多，骨折愈合快，抗弯应力强度大；肌内诱导成骨明显。酒精胎骨混悬液注射移植是一种简而有效的植骨材料和方法。     

异种移植动物模型-转人CD59 cDNA首建小鼠的培育

目的　培育转人CD5 9cDNA首建小鼠 ,以进一步研究异种移植 ,并为建立鼠系奠定基础。方法　构建转基因结构pEGFP -C1-OMT -CD5 9,限制性内切酶切除质粒载体 ,回收、纯化包括人CD5 9cDNA、绿色荧光蛋白(greenfluorescentprotein ,GFP)基因及各自启动子的约 3 .3kb的基因片段 ,用显微注射法将此片段注入小鼠受精卵 ;提取F0 代小鼠尾组织DNA ,用PCR法初选 ,再用Southern杂交法对PCR阳性鼠进一步筛选 ,确定阳性转基因首建小鼠。结果　共注射 5 90枚受精卵 ,移植于 2 4只受体母鼠中 ,出生F0 代小鼠 73只 ;PCR初选有 8只呈阳性(6♂∶2♀ ) ,Southern杂交有 2只呈阳性 (1♂∶1♀ ) ;经粗略估算 ,整合外源基因拷贝数分别为 3和 10。结论　成功地培育出转人CD5 9cDNA首建小鼠 ,用PCR和Southern杂交法证实了外源片段的整合     

Gynandroblastoma of ovary with juvenile granulosa cell component and heterologous intestinal type glands

An ovarian gynandroblastoma in a 15-year-old girl is described. The predominant component was juvenile granulosa cell tumour. Areas of adult granulosa cell tumour and Sertoli cell elements were also present. Stromal theca and luteinised cells were identified. An additional histological finding was the presence of heterologous intestinal type glands. There was positive immunohistochemical staining of juvenile and adult granulosa cell areas with inhibin and MIC2 antibodies. Electronmicroscopy showed a close ultrastructural resemblance between tumour cells in granulosa and Sertoli cell areas, in spite of differences in architectural pattern, suggesting that both morphological components may derive from a single cell of origin. The tumour demonstrates a unique combination of elements which has not previously been described.     

创面异体移植培养表皮细胞后存活的判定

应用４种方法判定烧伤创面异体移植的培养表皮细胞存活情况：１．聚合酶链反应（ＰＣＲ）查Ｙ染色体特异性ＤＮＡ片段；２．ＰＣＲ扩增ＰＭＣＴ１１８长度多态性基因位点；３．ＰＣＲ／ＳＳＯＨＬＡ－ＤＱａ分型；４．ＤＮＡ指纹。共检测１６例病人的２７个活检标本，均证实有供者的ＤＮＡ，最长时间为移植后９２天。初步实验结果表明：培养表皮异体移植后可存活较长时间。