An ultrastructural,immunohistochemical, and cytogenetical study of a monophasic pulmonary synovial sarcoma: implications of the frequent ultrastructure of oligocilia and concentric membranous bodies with positive immunostaining for VS38c and MEF2

A rare case of a monophasic pulmonary synovial sarcoma is reported. A 44-year-old Japanese man underwent lower lobectomy for a nodular mass in his right lung. Immunohistochemical study of the excised primitive spindle cell sarcoma revealed occasional positive stains by hitherto reported antigens of S-100, cytokeratin 7, high molecular weight cytokeratin (34betaE12), pankeratin (AE1/AE3), and EMA, which were helpful for the differential diagnosis of other spindle cell sarcomas. Furthermore, positive immunostains for MEF2, VS38c (plasma cell antigen), and bcl-2 were rather significant findings in the present case. The definitive evidence that molecular genetic analysis showed a clonal single electrophoretic band of SYT-SSX mutated chimera gene was conclusive for the pathological diagnosis. The implications of the frequently seen ultrastructure of oligocilia and concentric membranous bodies with positive stains for VS38c and MEF2 are discussed. In the difficult pathological diagnosis of a rare and undifferentiated type of sarcoma with unusual clinicopathological features generated at an unusual site, comprehensive ultrastructural, immunohistochemical, and cytogenetic studies will lead to the correct pathological diagnosis and elucidate the detailed characteristics of the tumor.     

小鼠→大鼠异种移植耐受模型的建立

目的：用非清髓方法建立小鼠→大鼠混合嵌合体模型，探讨免疫耐受机制。方法：给SD大鼠亚致死全身照射(TBI)后，4h内输入Balb／c小鼠骨髓细胞(BMC)，2d后腹腔注射环磷酰胺(CTX)，分别于BMT后30、60和90d，检测小鼠源性BMC在大鼠体内植活情况。通过皮肤移植、迟发超敏反应(DTH)和混合淋巴细胞反应(MLR)检查，探讨其耐受机制。结果：经处理大鼠外周血可测出小鼠源性嵌合体，皮肤移植、DTH和MLR检查显示对Balb／c小鼠产生特异性耐受，且较持久。结论：应用7．5Gy TBI 腹腔注射50mg／kg CTX 供体BMT、可成功建立小鼠→大鼠混合嵌合体模型诱导特异性耐受，嵌合体与耐受有关系。     

嵌合体大鼠诱导异种皮肤移植免疫耐受的初步研究

目的 探讨嵌合体大鼠诱导异种皮肤移植免疫耐受的可行性。方法 采集兔骨髓干细胞，经行宫内胎鼠移植以及对新生子鼠行肝内注射，制作兔骨髓干细胞嵌合体大鼠模型。6周后将15只嵌合体大鼠分为A组（8只）、B组（7只）。将A组大鼠皮肤移植给供髓兔，将7只非嵌合体大鼠皮肤移植给非供髓兔作A组对照；将B组大鼠、7只非嵌合体大鼠（B组对照）皮肤同时移植给供髓兔。记录移植后皮片成活时间、创面愈合时间。结果A组对照移植皮片成活（9．3±1．8）d，创面于（20．9±2．1）d愈合；A组移植皮片成活（15．1±2．6）d，创面于（18．5±1．3）d愈合。B组及其对照移植皮片的成活时间、创面愈合时间与A组相似。与各自对照皮肤移植相比，A、B组皮片成活时间延长（P〈0．01），创面愈合时间缩短（P〈0．05或P〈0．01）。结论 嵌合体大鼠行异种皮肤移植后能诱导移植皮片免疫耐受．使其成活时间明显延长．创面愈合时间缩短。     

联体共生诱导大鼠心脏移植免疫耐受的实验研究

目的　通过联体共生模型 ,建立供、受者嵌合体 ,探讨嵌合体与免疫耐受的关系。方法　纯系雄性DA(RT1a)大鼠为供者 ,Lewis(RT11)大鼠为受者 ,随机分成 3组 ,每组供、受者各 15只。Ⅰ组 (未处理组 ) :仅行DA到Lewis大鼠的腹部心脏移植 ,手术前后不作任何处理。Ⅱ组 (环磷酰胺组 ) :DA到Lewis大鼠的心脏移植前后分别经腹腔注射环磷酰胺 80mg/kg。Ⅲ组 (联体组 ) :0d :供、受者大鼠腹腔注射环磷酰胺 80mg/kg ;第6d :供、受者联体 ;第 16d :联体大鼠腹腔注射环磷酰胺80mg/kg ;第2 1d :分开联体 ,行DA到Lewis大鼠的心脏移植。观察各组移植心存活时间 ,供心病理学改变 ,供、受者间的混合淋巴细胞反应 (MLR)。结果　Ⅲ组形成了稳定的供、受者嵌合体 ,供心平均存活时间为 :(76 .33± 10 .71)d ,较Ⅰ组 (7.17± 1.17)d、Ⅱ组 (8.5 0± 1.87)d显著延长 ,差异有显著性 (P <0 .0 1) ;Ⅲ组的供心仅见少量炎性细胞浸润 ;供、受者间MLR较正常对照组显著降低 ,差异有显著性 (P <0 .0 1)。结论　联体共生可形成稳定的外周和中枢嵌合体 ,嵌合体在同种心脏移植的免疫耐受中起重要作用。     

一例PCDH19基因新生突变男性嵌合体致癫痫临床特点分析并文献复习

PCDH19基因突变主要引起限于女性的癫痫伴智力低下。因其特殊的遗传方式主要导致女性杂合子发病。目前PCDH19突变女性病例报道已达200余例,其临床特点已较明确。目前PCDH19基因突变男性患者报道仍然较少。其发病临床特点尚不十分清楚,亟待更深入的研究。本文报道1例PCDH19基因致病性突变男性嵌合体患儿的临床资料,基因检测结果提示PCDH19 c.1078（exon 1）G>A（p.Glu360Lys）变异,为新生突变,拓宽了PCDH19基因突变致病的基因表型谱。经过文献检索分析既往报道的13例性染色体核型为XY型的男性PCDH19基因致病性突变男性嵌合体患儿临床资料。14例男性嵌合体患儿（包含本文病例）发病特点为：（1）均有癫痫发作,100%呈丛集性发作,78.6%（11/14）具有热敏感性,发作起始年龄为5~31个月,中位年龄9个月,首次发作多为全面性发作,强直阵挛发作常见,后期以局灶性发作常见,35.7%（5/14）病程中存在癫痫持续状态;（2）癫痫发作前多数智力正常,癫痫发作后76.9%（10/13）存在智力障碍和精神症状。本文为临床诊治PCDH19基因突变患儿提供一定的理论依据。     

诱导持久稳定的嵌合体维持异种耐受的研究

目的　探讨诱导持久稳定的嵌合体来维持耐受的方法。方法　给SD大鼠亚致死照射 +BALB/c小鼠骨髓细胞 +环磷酰胺建立混合嵌合体模型后 ,于BMT后 14d输注供鼠脾细胞 3× 10 7或于 14 ,2 1,2 8d输注供鼠脾细胞 1×10 7、3× 10 7、5× 10 7,在输注脾细胞后 90 ,12 0d ,检测小鼠源性细胞在受鼠体内植活情况 ,通过混合淋巴细胞反应 (MLR)及迟发性超敏反应 (DTH)检测诱导维持耐受情况。结果　模型建立后未输注组 ,受鼠外周血于 90d时可测出小鼠源性细胞 ,但 12 0d已测不出 ,MLR和DTH检测显示耐受已消失 ,而输注组 12 0d时受鼠外周血仍可测得小鼠源性细胞 ,MLR及DTH检查仍出示特异性耐受 ,且BMT后第 14 ,2 1,2 8天输注组和 14d输注组 ,差异有显著性。结论　输注供体脾细胞可成功诱导持久稳定的混合嵌合体来维持特异性耐受。     

A combination of mutations enhances the neurotropism of AAV-2

There is strong interest in developing practical strategies for gene delivery to the central nervous system (CNS). Direct delivery into the brain or spinal cord is highly invasive as well as inefficient or hazardous using most current vector systems. Our objective was to generate innocuous gene vehicles that would be effectively taken up by axons and then home to the neuron cell bodies. Vectors derived from Adeno-Associated Virus (AAV), a harmless human parvovirus, offer strong starting candidates for deriving such vehicles. Enhancing the axonal uptake of AAV, and conferring more efficient retrograde transport capabilities upon the virus, should produce near ideal gene transfer vehicles for the CNS. To enhance retrograde transport of the virus, peptides mimicking binding domains for cytoplasmic dynein were inserted in the capsid by directed mutagenesis. In separate clones, peptides derived from an NMDA receptor antagonist were also introduced to provide a specific affinity for this receptor. When combined, these two functionally distinct classes of mutation enabled efficient gene transfer into neurons under conditions not permissive for standard AAV-2 vectors prepared under the same conditions. These results hold strong promise for the development of safe, convenient vehicles to target genes and other sequences to neurons, enabling new and novel approaches for the treatment of multiple neurological disorders.     

Cell lineage in fracture healing in chimeric bone grafts

Summary Previous studies have shown that differences in nuclear morphology are generally sufficient to determine the species origin of cells in interspecific grafts between the Japanese quail and domestic chicken. Most quail nuclei possess 1–3 large nucleolus-associated masses of heterochromatin. Chick cells, on the other hand, usually present a more diffuse, stippled distribution of nuclear heterochromatin. Quail embryonic limb rudiments, some with and some without established marrow cavities, were explanted and grown on the chorioallantoic membrane of the chick. Three to five days post-grafting, the explants were surgically fractured and allowed to heal. Tissues were collected and histologically processed during the latter period. The fractures healed completely within 5–6 days and no callus was established in the process. The nuclear staining pattern of the osteoblasts and osteocytes throughout the rudiments and at the fracture site indicated that they were derived from the graft. Possible sources for these cells included the periosteum, endosteum, and posthypertrophy chondrocytes. By contrast, most of the nuclei in the osteoclasts were chick-like and were apparently derived from cells originating in the host. Because the quail-like heterochromatin marker was normally present in a small number (2.5%) of chick osteoclast nuclei and was lacking in about 5% of native quail osteoclast nuclei, the precise extent of the participation of donor, i.e., quail bone and marrow stromal cells in osteoclast formation, could not be determined. However, the data suggest that in large measure the precursor cells for most osteoclasts were hematogenously derived and were carried to the grafted rudiments by the blood vascular system.