骨髓间质干细胞体外分化为成骨细胞的实验研究

建立猪骨髓间质干细胞 (mesenchymalstemcells ,MSCs)体外分离培养方法。对猪MSCs体外分化为成骨细胞的能力进行研究。抽取猪骨髓 ,体外培养MSCs。取第二代MSCs ,以含有不同浓度的抗坏血酸、β -磷酸甘油、地塞米松及碱性成纤维细胞生长因子等条件培养基进行成骨细胞诱导分化。通过细胞形态变化 ,碱性磷酸酶染色及钙盐沉积对成骨细胞进行鉴定。结果表明MSC细胞形态由长梭形向多边形转变 ,ALP染色阳性 ,VonKossa染色阳性 ,经体外诱导分化后呈典型的成骨细胞样改变。猪骨髓MSCs可在体外长期、稳定培养 ,具有向成骨细胞分化的潜能 ,可以为骨组织工程研究提供较理想的细胞来源和动物模型。     

腭部恶性肿瘤的CT诊断

探讨腭部恶性肿瘤的CT表现及其诊断价值。回顾性分析32例经病理证实的腭部恶性肿瘤的CT表现,其中男27例,女5例。所有病例均行横断增强扫描,其中8例同时行平扫,2例加冠状位扫描。腭部恶性肿瘤的CT表现为:(1)腭部肿块(26例)、腭部软组织增厚(6例)和腭骨质破坏(6例),其中病变位于软腭(22例)、硬腭(4例)、软硬腭交界处(3例);(2)邻近结构受累及(27例);(3)颈部淋巴结转移(17例)。CT是一种优良的检查技术,能显示腭部恶性肿瘤的大体病理改变以及侵犯途径,为临床治疗和预后提供重要信息。     

天然角蛋白自身反应性B细胞亚群及功能的分析

目的:明确天然角蛋白反应性B细胞的亚群及解剖定位,初步分析其分泌天然抗角蛋白自身抗体(anti-keratinautoantibody,AKautoAb)的能力。方法:取SPF级C57BL/6小鼠的脾细胞和腹腔细胞,荧光抗体染色后用流式细胞仪分析角蛋白反应性B细胞亚群。将脾脏和腹腔淋巴细胞体外培养后,用ELISA分析AKautoAb的滴度,用ELISPOT法分析分泌AKautoAb的B细胞数。结果:腹腔中几乎所有结合角蛋白的B细胞均为B-1a细胞,脾脏中结合角蛋白的B细胞以边缘带B细胞为主。腹腔细胞中分泌AKautoAb的细胞数显著多于脾细胞,其培养上清中AKautoAb的滴度显著高于脾细胞。结论:角蛋白反应性B细胞存在于3个成熟B细胞亚群:B-1细胞、滤泡B细胞和边缘带B细胞,其中CD5 的B-1a细胞具有活跃的分泌AKautoAb的能力。     

应用IRS-PCR对金黄色葡萄球菌分型的研究

目的探讨低频限制性位点聚合酶链反应(infrequent-restriction-site PCR，IRS-PCR)在金黄色葡萄球菌(简称金葡菌)基因分型中的应用价值。方法建立本实验室IRS-PCR方法。同时用IRS-PCR和脉冲场凝胶电泳(PFGE)对金葡菌进行基因分型。根据49株社区感染分离菌的分型结果计算辨别力指数(ID)值估计分辨力。对其中30株社区感染菌重复实验一次估计重复性。比较两种基因分型方法的分型率、分辨力、重复性、结果的一致率及操作特点。结果建立IRS-PCR对金葡菌基因分型的方法。70株金葡菌均可被2种方法分型，分型率100％。IRS-PCR分为38个型，21株院内感染菌分为6个型，49株社区感染菌分为32个型，计算ID值为0．981。PFGE分为40个型，21株院内感染菌分为6个型，49株社区感染菌分为34个型，计算ID值为0．983。两种分型方法的重复性均为100％。对院内感染菌，两种方法分型的一致率为100％；对社区感染菌，两种方法分型的一致率为92％(45／49)。与PFGE相比，IRS-PCR更简单、省时、易于操作、不需特殊昂贵仪器。结论IRS-PCR能对金葡菌简易快速可靠分型，适合检验科对临床标本的快速有效分型，是一种有价值的分子流行病学研究工具。     

因特网上医学影像学资源获取技术

目的探讨医学网络资源的一般获取技巧,以及医学影像学资源的获取途径。方法应用因特网等关键词,采用医学专业检索引擎Medical Matrix,并在此基础上链接其他医学检索引擎,获得有关医学影像学文献的网址和相关网页。结果网上有一些专门的医学影像学资源网站,在高等医学院校和医学科研机构中经常包含医学影像学的资源。结论本文具体阐述了医学影像学资源的获取途径和技巧。     

上矢状窦旁桥静脉的显微解剖与影像学观察

目的:通过对上矢状窦(SSS)旁大脑桥静脉的显微解剖、影像学观察及其对照研究,为各种纵裂间手术入路中桥静脉的保护提供基础。方法:分别对30例(60侧)上矢状窦和颈内静脉内灌注蓝色乳胶的成人头颅湿标本、40侧DSA静脉相和16侧CTV进行显微解剖和影像学观察。结果:上矢状窦前部和后部分别有一缺乏桥静脉注入段,显微解剖、DSA和CTV观察上矢状窦旁桥静脉的数量分别为11.2支、8.9支和7.0支,DSA和CTV观察发现桥静脉注入上矢状窦处常显示不清。结论:熟悉和比较上矢状窦旁桥静脉的解剖学和影像学特征有利于各种纵裂间手术入路和手术过程中桥静脉的保护。     

人类白细胞抗原-G突变体cDNA克隆及在K562细胞上的表达

目的：克隆人类白细胞抗原-G(Human leukocyte antligen-G，HLA-G)突变体cDNA，并使其在HLA-I类阴性的K562细胞上获得稳定表达，为研究配-受体之间的识别机制奠定基础。方法：用RT-PCR方法从人子宫蜕膜组织扩增出HLA-GcDNA，得到全长HLA-GPCR产物后，用桥式PCR方法进行定点点突变，将突变的目的基因亚克隆于逆转录，将突变的目的基因亚克隆于逆转录mG-pLNCX表达载体，采用感染的方法将重组质粒转入K562细胞，最后经G418筛选及有限稀释，利用单克隆抗体W6/32进行FACS及mRNA检测，观察HLA-G突变体在靶细胞表面的表达。结果：HLA-G突变体分子在经mG-pLNCX转染的靶细胞表面获得稳定高表达。结论：成功构建了mG-pLNCX表达载体，并使HLA-G突变体分子在HLA-I类阴性的靶细胞K562细胞上获得稳定表达。     

肾混合性上皮间质肿瘤和成人囊性肾瘤的临床病理学观察

Objective To study the clinicopathologie features,immunophenotype and differential diagnosis of mixed epithelial and stromal tumor of kidney ( MEST) and adult cystic nephroma ( CN).Methods Five cases of MEST and 4 cases of CN were retrospectively analyzed.Immunohistochemical study was carried out and the literature was reviewed.Results All of the five patients with MEST were females.Their median age was 45 years.For CN,there were 3 males and 1 female and their median age was 41 years.All patients presented with loin pain and hematuria.On gross examination,MEST was well-circumscribed but non-encapsulated.There was no evidence of haemorrhage or necrosis.Three of the cases were solid in nature.One was composed of a mixture of solid and cystic elements,while the remaining case showed a multicystic cut surface bridged by thick fibrous septa.On the other hand,CN were well-circumscribed and encapsulated.They were multiloculated cystic in nature.The cystic spaces were separated by thin septa and there was no significant solid or necrotic component.Histologically,MEST consisted of proliferation of cystically dilated glands admixed with spindly stromal cells with various cellularity and growth patterns.Both the glandular and stromal elements were well-differentiated with no cytologic atypia identified.The glandular structures in 2 of the cases were partially lined by endometrial or tubal epithelium.In contrast,the thin-walled cystic spaces in CN were lined by a single layer of epithelium.Immunohistochemical　study showed that the epithelial cells were positive for pan-cytokeratin and epithelial membrane antigen.The spindle cells in MEST expressed vimentin (5/5 ) ,smooth muscle actin (3/5 ),desmin (4/5 ),CD10 (5/5),estrogen receptor (4/5) and progesterone receptor (4/5).They were negative for HMB45,CD34,CD117 and S-100 protein.On the other hand,the spindle cells in CN were variably positive for vimentin (4/4),smooth muscle actin (4/4),desmin (1/4),estrogen receptor (3/4) and progesterone receptor (1/4).They were negative for CD10,HMB45,CD34,CD117 and S-100 protein.Conclusions Both MEST and CN are uncommon renal neoplasm.Most of them run a benign clinical course.The stromal cells in MEST show smooth muscle or myofibroblastic differentiation.Areas demonstrating Miillerian features also existed in some cases.MEST and CN share overlapping histological and immunohistochemical features,and may represent spectrum of the same group of lesions.     

Parkin localizes to the Lewy bodies of Parkinson disease and dementia with Lewy bodies

Mutations in alpha-synuclein (alpha S) and parkin cause heritable forms of Parkinson disease (PD). We hypothesized that neuronal parkin, a known E3 ubiquitin ligase, facilitates the formation of Lewy bodies (LBs), a pathological hallmark of PD. Here, we report that affinity-purified parkin antibodies labeled classical LBs in substantia nigra sections from four related human disorders: sporadic PD, inherited alphaS-linked PD, dementia with LBs (DLB), and LB-positive, parkin-linked PD. Anti-parkin antibodies also detected LBs in entorhinal and cingulate cortices from DLB brain and alphaS inclusions in sympathetic gangliocytes from sporadic PD. Double labeling with confocal microscopy of DLB midbrain sections revealed that approximately 90% of anti-alpha S-reactive LBs were also detected by a parkin antibody to amino acids 342 to 353. Accordingly, parkin proteins, including the 53-kd mature isoform, were present in affinity-isolated LBs from DLB cortex. Fluorescence resonance energy transfer and immunoelectron microscopy showed that alphaS and parkin co-localized within brainstem and cortical LBs. Biochemically, parkin appeared most enriched in cytosolic and postsynaptic fractions of adult rat brain, but also in purified, alpha S-rich presynaptic elements that additionally contained parkin's E2-binding partner, UbcH7. We conclude that parkin and UbcH7 are present with alphaS in subcellular compartments of normal brain and that parkin frequently co-localizes with alpha S aggregates in the characteristic LB inclusions of PD and DLB. These results suggest that functional parkin proteins may be required during LB formation.     

Ⅲ型胶原肾小球病的形态学观察

目的了解Ⅲ型胶原肾小球病的形态学改变,并对Ⅲ型胶原可能的细胞来源进行初步探讨。方法对3例肾活检组织进行光镜、免疫荧光、电镜和Ⅰ、Ⅲ、Ⅳ型胶原及d平滑肌肌动蛋白(α-SMA)的免疫组织化学染色(SP法)观察。结果2例患者临床表现为肾病综合征,其中1例伴高血压,第3例表现为肾功能不全和肾性高血压。3例均无肾病家族史。光镜检查可见肾小球基膜内和系膜区弥漫性过碘酸-希夫反应阳性物质沉积,系膜细胞无明显增生。电镜检查在基膜内疏松层和系膜区可见大量胶原纤维沉积,系膜细胞胞膜下平行排列的束状微丝明显增加。免疫组织化学显示这些胶原纤维为Ⅲ型胶原,Ⅰ型和Ⅳ型胶原阴性,同时系膜区多数系膜细胞α-SMA阳性。结论Ⅲ型胶原肾小球病光镜、电镜及免疫组织化学上都有其特殊的病理改变。肾小球内激活的系膜细胞可能是Ⅲ型胶原的来源。