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。     

Size fractions of ambient particulate matter induce granulocyte macrophage colony-stimulating factor in human bronchial epithelial cells by mitogen-activated protein kinase pathways

Environmental pollutants, including ambient particulate matter (PM), increase respiratory morbidity. Studies of model PM particles, including residual oil fly ash and freshly generated diesel exhaust particles, have demonstrated that PM affects inflammatory airway responses. Neither of these particles completely represents ambient PM, and therefore questions remain about ambient particulates. We hypothesized that ambient PM of different size fractions collected from an urban environment (New York City air), would activate primary culture human bronchial epithelial cells (HBECs). Because of the importance of granulocyte-macrophage colony-stimulating factor (GM-CSF) on inflammatory and immunomodulatory processes, we focused our studies on this cytokine. We demonstrated that the smallest size fraction (ultrafine/fine; < 0.18 micro m) of ambient PM (11 micro g/cm(2)), upregulated GM-CSF production (2-fold increase). The absence of effect of carbon particles of similar size, and the day-to-day variation in response, suggested that the chemical composition, but not the particle itself, was necessary for GM-CSF induction. Activation of the extracellular signal-regulated kinase and the p38 mitogen-activated protein kinase was associated with, and necessary for, GM-CSF release. These studies serve to corroborate and extend those on model particles. Moreover, they emphasize the role of the smallest size ambient particles in airway epithelial cell responses.     

3～20周人胚和胎儿肾小体的组织发生

应用光、电镜对3～20周22例人胚和胎儿肾小体的组织发生进行了观察.受精后第25天胚肾已有肾小体发生.随着胚龄的增长,肾小体的发生数目增多,肾小体的形成方式是:胚肾内先形成许多厚壁小管,在一部分厚壁小管的诱导下,另一部分厚壁小管的一侧壁呈帽状增厚,分化形成造血干细胞、毛细血管内皮及肾小囊脏层,对侧壁形成肾小囊壁层.即厚壁小管的双侧壁形成肾小体.     

Heterogeneity of hepatitis delta antigen

Hepatitis delta antigen (HDAg) is the only known protein encoded by the hepatitis delta virus (HDV). Two HDAg species of different sizes have been detected in the sera and livers of the infected humans, chimpanzees, and woodchucks, even though only one RNA species was previously identified in most of the HDV strains. To study HDAg heterogeneity, we took advantage of the fact that a single base mutation at nucleotide 1015 (C to U), which results in an amber termination codon in the HDAg open reading frame (ORF), eliminates a unique Ncol restriction enzyme site. We screened various HDV cDNA clones and detected sequence heterogeneity of the HDAg-coding region on the basis of the presence or absence of the Ncol site. Five delta hepatitis patients were examined. In every patient, two types of HDAg-coding sequence were detected at nucleotide 1015: one which contains a C and results in an ORF encoding a delta antigen of 214 amino acids, and the other which possesses a U and results in an amber termination codon and a truncated HDAg species of 195 amino acids. The in vitro translation products of these two ORFs comigrated with the two HDAg species from the patient's plasma on SDS polyacrylamide gels. Polymerase chain reaction (PCR) amplification of the HDV RNA from some patients' sera and subsequent sequencing showed several additional mutations in the HDAg-coding region. These mutations are independent of the C or U nucleotide change at the site of the amber termination codon.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of immunohistochemical markers in the differential diagnosis of adrenocortical tumors: immunohistochemical analysis of adrenocortical tumors.

Most adrenocortical tumors (ACTs) can be diagnosed directly by a combination of morphologic features and clinical findings. However, sometimes it may be difficult to distinguish ACTs from other neoplasms such as pheochromocytomas and some metastatic tumors, particularly for small biopsy specimens because they may be morphologically similar. Expression of calretinin has recently been suggested as a valuable immunomarker for the differential diagnosis between ACTs and other tumors; however, its diagnostic value is still under debate. To determine the diagnostic value of calretinin in Chinese patients with adrenocortical and non-ACTs, we employed both polyclonal and monoclonal anticalretinin to characterize the expression of calretinin in adrenal tissues and compared its expression with that of inhibin alpha, Melan-A, cytokeratin, or CD99 by immunohistochemistry in tissue microarrays and standard tissue sections of 414 specimens. Our results revealed that calretinin was expressed by adrenocortical cells, but not by the other cells tested and the percentage of calretinin-positive ACTs reached 99% when stained with polyclonal antibodies, which was higher than that with monoclonal anticalretinin (91.3%), anti-Melan-A (90.3%), antiinhibin alpha (81.6%). In addition, our results also revealed that ACTs were stained by cytokeratin (AE1/AE3) with variable degrees (58.7%). Furthermore, unlike anti-Melan-A that stained all metastatic malignant melanoma, anticalretinin did not recognize other tested tumors. Therefore, immunohistologic staining with polyclonal anticalretinin is more sensitive than other antibodies tested for the diagnosis of ACTs. However, monoclonal anticalretinin appeared to be more specific. Importantly, our data suggested that the fried-egg-like staining pattern, but not the mere cytoplasmic staining, was characteristic of anticalretinin staining in adrenocortical tissues. Notably, a few anticalretinin negative-ACTs were stained by other immunomarkers that we tested. Thus, the combinational characterization of calretinin (either by polyclonal or monoclonal antibody), inhibin alpha, and Melan-A expression is of great significance in the differential diagnosis of ACTs.     

兔高动力性肺动脉高压模型的建立

目的： 研究通过幼兔长时间体－肺循环分流，建立高动力性肺动脉高压模型方法的可行性。 方法： 将1月龄幼兔正中开胸，行左颈总动脉与主肺动脉吻合，形成持续左向右分流。3个月后通过彩超证实吻合血管通畅性，并测定其肺动脉收缩压（PASP）、肺动脉舒张压(PADP)、平均压(MPAP)，观测肺小动脉病理变化、管壁厚度指数(TI)、面积指数（AI）。 结果： 分流组3个月后，21只形成肺动脉高压。分流血管阻断前、后PASP、PADP、mPAP明显高于对照组（P＜0.05）。肺组织病理检查示肺小动脉管壁增厚、管腔狭窄，与对照组比，TI和AI 明显增加（P＜0.05）。 结论： 幼兔经体－肺循环分流手术3个月后，可形成与临床先心病病理生理相接近的高动力性肺动脉高压。该模型稳定、可靠、经济。     

不同胎龄的胎儿和少儿皮肤中bax,bcl-2和p53基因表达的变化

目的：探讨凋亡相关基因bax, bcl-2和p53在不同胎龄的胎儿皮肤和少儿皮肤组织中表达的变化特征及其可能的生物学意义。方法： 运用末端脱氧核糖转移酶介导的生物素化脱氧尿嘧啶缺口标记技术(TUNEL)检测18例不同胎龄(13-32周)的胎儿皮肤和6例少儿皮肤组织中细胞凋亡的变化后，提取这些皮肤组织中的总RNA，分离mRNA，用RT-PCR方法检测bax, bcl-2和p53基因在不同组织中的表达变化特征。结果： 随着胎儿的生长发育，皮肤组织中的细胞凋亡率逐渐增加。在早期妊娠胎儿的皮肤中，bcl-2基因表达水平较高，随着胎龄的增加，bcl-2基因的转录本含量逐渐降低，在少儿的皮肤组织中，这种基因的表达量明显低于早期妊娠胎儿皮肤（P<0.01）。与bcl-2基因不同，在早期妊娠胎儿皮肤组织中，p53基因表达水平较低，而在晚期妊娠胎儿和少儿的皮肤内，该基因表达较强，而bax基因在不同发育时期的胎儿和少儿皮肤组织中表达差异不显著（P>0.05）。结论： 晚期妊娠胎儿和少儿皮肤组织中细胞增殖减缓，细胞趋向分化或凋亡的增加可能与p53基因表达增强，bcl-2表达降低相关；而p53表达降低，bcl-2表达升高可能是早期妊娠胎儿皮肤中细胞凋亡较少的机制之一。     

Opa相互作用蛋白5在胰腺癌中的表达及其对PANC-1细胞增殖的影响

目的探索Opa相互作用蛋白5(OIP5)在胰腺癌中的表达及其对PANC-1细胞增殖的影响。方法通过数据库分析OIP5在胰腺癌组织及癌旁组织中的表达;用实时定量PCR(RT-qPCR)和蛋白印迹法(Western blot)分别检测人胰腺癌细胞系MIAPaCa-2、PANC-1、KP-3、BxPC-3细胞中OIP5 mRNA和蛋白表达;构建OIP5基因沉默质粒的慢病毒(pGCSIL-shOIP5)和对照质粒慢病毒(pGCSIL-shCtrl),分别感染PANC-1细胞,分为OIP5基因沉默组和shCtrl对照组,5 d后采用RT-qPCR和Western blot测定慢病毒敲低效率,流式细胞计量术检测细胞凋亡;OIP5基因沉默组和shCtrl对照组连续5 d进行MTT检测和细胞计数;OIP5基因沉默组和shCtrl对照组孵育10 d形成集落,Giemsa染色分别集落总数。结果胰腺癌中OIP5 mRNA表达显著高于正常胰腺组织(P<0.05),OIP5高表达患者的总存活率显著低于OIP5低表达患者(P<0.05),且其无病生存率也显著降低(P<0.05);OIP5在MIAPaCa-2、PANC-1和KP-3中表达较高,而在BxPC-3细胞系中的表达较低;MTT检测结果显示OIP5沉默在第4和第5天显著降低了PANC-1细胞的增殖速率(P<0.01);OIP5沉默后细胞集落数(平均为9个)显著低于shCtrl对照组中的数量(平均为40个)(P<0.01);OIP5沉默后PANC-1细胞凋亡比例为8.3%显著高于shCtrl的4.5%(P<0.01)。结论OIP5在胰腺癌细胞系中异常高表达,OIP5基因可调控胰腺癌PANC-1细胞的增殖、凋亡以及集落形成,提示OIP5可能在胰腺癌发病机制中作为癌基因发挥作用,从而为胰腺癌的靶向治疗提供了潜在的生物标志物。     

细胞角蛋白19、galectin-3、HBME-1在甲状腺病变上的表达及鉴别诊断意义

目的 研究细胞角蛋白(CK)19、galectin(Gal)-3、HBME-1在甲状腺不同病变表达的特点及鉴别诊断中的应用价值。方法 应用免疫组织化学EnVision法检测了21例结节性甲状腺肿(结甲)、14例毒性甲状腺肿(甲亢)、15例甲状腺滤泡性腺瘤(腺瘤)、13例滤泡性癌、13例滤泡型乳头状癌及48例经典型乳头状癌中单克隆抗体CK19、Gal-3、HBME-1的表达。结果 甲状腺病变中3种标记表达均位于细胞质;CK19、Gal-3、HBME-1的表达在甲状腺良性病变(结甲、甲亢、腺瘤)大多为弱阳性或阴性,而滤泡性癌阳性明显增加、乳头状癌(滤泡型及经典型)大多为中、强阳性,3种标记在甲状腺不同病变的阳性表达率结甲为52．4％(11／21)、9．5％(2／21)、19．0％(4／21),甲亢为50．0％(7／14)、7．1％(1／14)、7．1％(1／14),腺瘤为60％(9／15)、13．3％(2／15)、13．3％(2／15),滤泡性癌为76．9％(10／13)、61．5％(8／13)、53．8％(7／13),滤泡型乳头状癌为：100％(13／13)、84．6％(11／13)、92．3％(12／13),经典型乳头状癌为100％(48／48)、93．8％(45／48)、95．8％(46／48);在甲状腺良性病变(结甲、甲亢、腺瘤)与恶性病变(滤泡性癌、乳头状癌)间3种标记差异均有显著性(P均=0．000);同时3种标记在滤泡样病变即腺瘤、滤泡性癌和滤泡型乳头状癌间亦有显著差异(CK19：P=0．038,Gal-3：P=0．001,HBME-1：P=0．000)。结甲有9例,甲亢有7例,腺瘤有6例3种标记均不表达,滤泡性癌仅有1例,而乳头状癌(滤泡型及经典型)没有病例3种标记均不表达,同一病例有2种以上阳性表达在结甲、甲亢、腺瘤、滤泡性癌、滤泡型乳头状癌和经典型乳头状癌中分别为14．2％(3／21)、21．4％(3／14)、20．0％(3／15)、69．2％(9／13)、92．3％(12／13)、100．0%(48／48),在甲状腺良性病变与恶性病变间以及滤泡样病变间差异亦有显著性(P=0．000)。结论 CK19、Gal-3、HBME-1的检测尤其是联合检测对甲状腺病变的诊断、鉴别诊断具有较高的实用价值。     

应用聚合酶链反应技术检测肝组织中的HBV－DNA

采用聚合酶链反应（ＰＣＲ）技术，对４２例肝活切组织石蜡切片中乙型肝炎病毒（ＨＢＶ）ＤＮＡ进行检测，并与乙肝表面抗原（ＨＢｓＡｇ）的免疫组织化学及血清学检测进行比较，ＨＢＶ－ＰＣＲ阳性率为７３．８％，高于组织及血清ＨＢｓＡｇ阳性率（分别为５９．５％和５０．０％）。３例病理形态呈肝炎改变，而血清ＨＢｓＡｇ（─）的肝组织中有２例检出ＨＢＶ－ＤＮＡ，提示ＰＣＲ的高度敏感性和准确性。８３．３％的门脉性肝硬变和８７．５％的肝细胞癌组织中ＨＢＶ－ＰＣＲ呈阳性，进一步证实了上述两病与ＨＢＶ的关系密切。我们还发现肝细胞淤胆患者ＨＢＶ感染率较高，ＨＢＶ－ＤＮＡ及组织ＨＢｓＡｇ阳性比例各为６／９和４／８。