Subunit composition of strychnine-sensitive glycine receptors expressed by adult rat basolateral amygdala neurons

In neonatal rats, strychnine-sensitive glycine receptors are widely expressed in the spinal cord, brainstem and forebrain. During development, these 'neonatal' receptors are replaced by an adult isoform, the expression of which becomes restricted primarily to brain stem and spinal cord. Unlike most forebrain regions, functional strychnine-sensitive glycine receptors appear to persist within adult rat amygdala. However, the subunit composition of glycine receptors expressed by amygdala neurons and its relationship to the adult isoform in brain stem/spinal cord has not been defined precisely. In this report, we have utilized RT-PCR and single-cell RT-PCR to demonstrate that the 'neonatal' alpha2-subunit mRNA persists in adult rat amygdala neurons and is the predominant alpha-subunit. We further demonstrate that native amygdala glycine receptors are relatively insensitive to the receptor antagonist picrotoxin, suggesting that alpha2- and beta-subunits may be present together in the same multisubunit complex. We further demonstrate that alpha2- and beta-subunits cloned from adult rat amygdala can form functional channels when expressed in a heterologous system. Together, these studies highlight both the unique characteristics of strychnine-sensitive glycine receptors in the adult rat amygdala as well as the possibility that alpha2/beta channels may represent the adult forebrain isoform of the strychnine-sensitive glycine receptor.     

接受供精辅助生殖技术治疗夫妇知情同意与生育伦理知识调查研究

目的：探讨接受供精辅助生殖技术（ART）治疗夫妇对知情同意和生育伦理的认知、行为和态度。方法：对在广东省计划生育专科医院登记接受供精人工授精技术治疗的92例对象进行自填式问卷调查。内容包括一般信息、生育压力、知情同意和生育伦理的认知、态度和行为。资料处理方法采用描述性统计。结果：对象面临强烈的生育渴求和巨大的生育压力,且主要来源于对象自己、社会和父母。对供精人工授精技术治疗有比较充分的知情,但仅有75%的对象知道ART后代可能发生出生缺陷。在接受供精人工授精技术治疗时,如果可以选择供精者生物学特征时,58%的对象优先选择供精者身高,其次是相貌和学历。对象支持在治疗前签署知情同意书。结论：对接受供精ART治疗的夫妇,在治疗前应做好知情同意的全过程。     

异种脱细胞真皮重建眼睑的实验研究

目的探讨异种（猪）脱细胞真皮基质（Xeno—ADM）植入兔眼睑后的组织相容性。方法38只健康新西兰白兔,随机平均分成实验组和对照组,均游离去除下眼睑5mm×4mm的全层睑板,分别植入Xeno-ADM和异体巩膜,观察大体情况,于术后1、2、4、6、8、12、16周分别取植片交界处,HE染色光镜下观察组织学变化,并同时取离心血清及植片组织匀浆上清液用放射免疫法检测白细胞介素2（IL-2）和白细胞介素6（IL-6）的含量。结果Xeno-ADM植入兔眼睑后,炎性反应轻,血管化及新生胶原长入快,植片与正常组织容合佳;术后16周测量所有动物（n=10）睑裂平均宽度及高度,实验组、对照组、正常组两两之间差异均无统计学意义（P〉0．05）。免疫学指标统计分析,植片组织匀浆液中IL-2和IL-6含量变化情况为：术后第1、2、4周升高不显著,第6～8周浓度明显升高,达峰值（实验组IL-2第6周为0．29281mg／L,IL-6第8周为118．258ng／L,对照组IL-2第8周为0．27799ng／ml,IL-6第8周为255．871pg／ml）,于12周降至极低水平,而第16周回升,其中对照组浓度可达第二峰值（IL-2为1．36341ng／ml,IL-6为622．863pg／ml）,明显高于第一峰值。外周血中IL-2和IL-6含量变化均呈波动性升高趋势,在植入后的4—8周有明显峰值,且与植入区域内炎性反应变化相似;应用重复测量方差分析发现,处理因素、时间因素对两组血清中IL-2和IL-6含量的影响无统计学意义（P〉0．05）。结论Xeno-ADM植入兔眼睑后组织相容性佳,可在局部及全身检测到高水平变化的IL-2和IL-6,其动态变化与同种异体巩膜相比,差异尚无统计学意义,考虑可替代巩膜重建眼睑,作为新的修复材料在眼科推广应用。     

大鼠坐骨神经去细胞基膜管异体移植的比较研究

目的:探讨化学萃取法(CEN)、冻融法(FTN)制备的异种鼠坐骨神经去细胞基膜管移植术后8、13、23天神经再生轴突距离、Schwann细胞迁入、再血管化及炎症反应异同.方法:将36只雌性Wistar鼠随机配对,分为CEN组与FTN组,每组18只,分别桥接移植两种方法制备的Sprague-Dawley鼠坐骨神经去细胞基膜管.观察术后8、13、23天移植物物理性状及组织形态学变化.结果:术后8、13、23天,神经轴突再生距离、近和远端Schwann细胞数、再血管化及炎症反应差异有显著性(P<0.05),CEN组优于FTN组.结论:化学萃取法制备的异种周围神经去细胞基膜管较冻融法能更快、更好地修复周围神经缺损,亦应有更大的临床实用价值.     

Adhesive functions of both chains of VLA-integrins are not fully conserved across the human-porcine species barrier: implications for xenotransplantation

BACKGROUND: A possible solution to the shortage of organs for transplantation would be the use of swine as source animals. As current immunosuppressive protocols cannot prevent rejection of these organs, super-selective immunosuppression or the induction of donor-specific central tolerance represent two promising approaches. Central tolerance induction involves bone marrow transplantation, and depends on intrathymic deletion of donor reactive host cells by donor antigen-presenting cells. In super-selective immunosuppression, the aim would be to block specific adhesive interactions on one species side only, leaving the other species side unaffected. As both processes depend on the interaction of adhesion molecules with their ligands, we investigated whether the beta1-integrins, which play roles in hematopoiesis as well as in rejection, can successfully interact across the swine-to-human species barrier. METHODS: We employed static cell-to-extracellular protein and cell-to-cell adhesion assays, using different cell types and monoclonal antibody as well as peptide-fragments to analyze conservation of cross-species adhesive interactions. RESULTS: We found that porcine and human cells interact differently with their cross-species ligands than their own and that the adhesive function of the beta1-chain does not seem to be fully conserved across the species barrier. CONCLUSIONS: Integrin functions are not fully conserved across the pig-to-human species barrier. While the development of multi-transgenic pigs, whose integrins interact with human ligands in a more 'human-like' manner may be necessary to facilitate tolerance induction, these facts give rise to new possibilities concerning super-selective immunosuppression.     

Evidence of macrophage receptors capable of direct recognition of xenogeneic epitopes without opsonization

BACKGROUND: We have previously demonstrated that porcine livers perfused with human blood remove most of the erythrocytes from three units of human blood over the course of a 72-h extracorporeal perfusion. Red blood cell loss did not appear to involve classical complement pathway-mediated hemolysis, but instead resulted from porcine Kupffer cell phagocytosis. METHODS: We developed a method incorporating collagenase digestion and metrizamide separation to isolate and maintain porcine Kupffer cells in primary culture. An in vitro rosetting assay was used to assess the binding of human and porcine erythrocytes to porcine Kupffer cells. Immunohistochemistry was used to confirm the presence of porcine macrophages. The rosetting assay was quantified using 51Cr-labeling of erythrocytes to assay for both rosette formation and phagocytosis. RESULTS: Porcine Kupffer cells were successfully isolated and maintained in primary culture. The presence of porcine macrophages was confirmed using the monoclonal antibody 74-22-15A. Human, but not porcine, erythrocytes were bound in an in vitro rosetting assay as confirmed by immunohistochemistry, electron microscopy and 51Cr-quantitation. Porcine Kupffer cells bound human erythrocytes regardless of the presence of opsonizing antibody. Approximately 70% of the isolated porcine Kupffer cells demonstrated the capacity to bind non-opsonized human erythrocytes. Phagocytosis was not observed. CONCLUSIONS: Using primary porcine Kupffer cell cultures, we have demonstrated that a subpopulation of porcine macrophages has the ability to recognize specifically xenogeneic human erythrocyte epitopes without the need for prior opsonization. The possibility is discussed that lectin-mediated carbohydrate binding plays a role in the cellular and humoral recognition and rejection of xenografts.     

异种板层角膜移植的免疫学及组织学观察

本实验以鸡对兔异种板层角膜移植为实验模型,动态观察手术前后3个月内角膜透明度、体液免疫、细胞免疫及组织学改变。结果发现术后3～4周异种角膜移植动物发生免疫排斥致植片混浊,3个月内又渐恢复透明。作者认为,细胞及体液因素均参与免疫排斥反应;受体角膜细胞的再生修复可能是植片又复透明的原因。     

Infantile Fibrosarcoma—a Misnomer?

Two cases of congenital or infantile fibrosarcoma are described that were incompletely excised at the time of primary excision and have not recurred or metastasized after 3 years. The tumors were composed of densely cellular spindle cells with a high mitotic index. Immunohistochemical stains were positive for vimentin but negative for desmin and S-100. The tumor cells were grown in vitro, and a karyotype was obtained. Both tumors had normal diploid modal karyotypes. In addition, fragments of the primary tumor from both cases were injected subcutaneously into nude mice; neither tumor could be heterotrans-planted. The clinical course and biologic features of these two tumors suggest that congenital or infantile sarcoma does not have the properties of a malignant neoplasm, and thus the designation of these tumors as a sarcoma may be a misnomer.     

Rhabdomyosarcoma of the cervix in sisters with review of the literature

OBJECTIVES: Pediatric embryonal heterologous rhabdomyosarcoma of the cervix is a rare tumor. METHODS: We present an interesting clinico-pathologic situation of two sisters presenting with pediatric embryonal heterologous rhabdomyosarcoma of the cervix. RESULTS: Pediatric embryonal heterologous rhabdomyosarcomas of the cervix are relatively uncommon. After a Pubmed search from 1952 to present, to our knowledge, this is the only report involving sisters presenting with this disease. Treatment has been extrapolated from collaborative groups such as Intergroup Rhabdomyosarcoma Study (IRS) Group so that optimal management may be achieved. CONCLUSIONS: We urge continued reporting of these rare tumors to enhance understanding if there may be a genetic component associated with them.     

非亲缘异基因骨髓移植治疗儿童白血病

目的　评价非亲缘异基因骨髓移植 (URD BMT)治疗儿童急性和慢性白血病的临床疗效。方法　 6例白血病患儿 ,其中慢性髓系白血病 2例 ,急性淋巴细胞白血病 3例 (第 1次完全缓解 ) ,急性早幼粒细胞白血病 1例 (第 2次完全缓解 ) ,由台湾慈济骨髓捐赠中心提供无关供者骨髓。预处理方案为马利兰 环磷酰胺 (Bu/Cy2 )方案 ,急性移植物抗宿主病 (aGVHD)预防为霉酚酸酯(MMF)、环孢菌素A(CsA)加氨甲喋呤 (MTX)联合方案 ;以前列素E1预防肝静脉闭塞病 (VOD) ,以巨细胞病毒 (CMV)抗原血症监测和更昔洛韦预防CMV病。供、受者间HLA基因位点型全相合 3例 ,1个基因位点型不合 2例 ,2个基因位点型不合 1例。结果　 6例患儿经DNA短串联重复序列多态性分析证明为供髓植入 ,中性粒细胞 >0 5× 10 9/L的中位天数为 14 5 (13～ 18)d ,血小板 >2 0×10 9/L的中位天数为 16 (11～ 2 3)d。发生Ⅱ～Ⅳ度aGVHD 2例 (33% ) ,局限性慢性移植物抗宿主病(cGVHD) 3例 ,未发生广泛性cGVHD。中位随访时间 4 12 (187～ 1338)d ,全部患儿均无病生存。结论非亲缘异基因骨髓移植是治疗儿童急性和慢性白血病的有效方法。