Restriction endonuclease fingerprinting enhanced conformation sensitive gel electrophoresis (REF-CSGE) in the analysis of BRCA1 exon 11 mutations in a high-risk breast cancer cohort

Efficient genetic analysis of large exonic regions containing heterozygous mutations and common polymorphisms can be difficult. We have analyzed 30 patients for inherited susceptibility mutations (ISM) within exon 11 of the BRCA1 gene as part of an ongoing genetic epidemiological study of high-risk breast cancer (HRBC). A novel combination of restriction endonuclease fingerprinting (REF) and conformation sensitive gel electrophoresis (CSGE) was developed for rapid and efficient screening of mutations. This method (REF-CSGE) was compared side-by-side with standard CSGE and evaluated for both efficiency and sensitivity of detection. REF-CSGE detected 100% of the alterations found by CSGE. However, one variant was only detectable by REF-CSGE. All samples with variant bands were sequenced to confirm the nature of the alteration. In total, two small deletions (frameshifts) and 62 point mutations (60 known polymorphisms and two variants of unknown significance) were found in our cohort. The majority of the exon 11 polymorphisms detected are inherited as a linked haplotype. Point mutations that comprise these haplotypes could be simultaneously detected on a single gel by REF-CSGE, thereby decreasing the number of sequencing reactions necessary to elucidate heteroduplex patterns seen on CSGE gels. An analysis of the overall efficiency of both techniques revealed that REF-CSGE required 67% fewer confirmatory sequencing reactions, resulting in savings in both reagents and technician time.     

镉对培养大鼠睾丸支持细胞的损伤及黄芪的拮抗作用

目的:探讨镉对培养大鼠睾丸支持细胞的损伤及黄芪的保护作用.方法:对照组和经镉、镉加黄苠处理的原代培养大鼠睾丸支持细胞用于波形蛋白、E-钙黏蛋白免疫组织化学检测、细胞内钙离子浓度测定及电镜观察.结果:镉处理后2种蛋白阳性产物表达较对照组减弱.镉加黄芪组阳性产物较对照组减弱但明显强于镉组.对照组、镉和镉加黄苠组细胞内钙离子浓度分别为(81.773±3.711)、(161.6024±1.978)和(104.278±1.963).镉处理后支持细胞超微结构受损明显,部分细胞核异染色质凝聚或核碎裂,胞质空泡化.镉加黄芪组细胞破坏较轻.结论:镉对培养支持细胞具有明显的毒性作用,而黄苠可以拮抗镉对支持细胞的损伤.     

虚拟咬合运动初步仿真模拟的研究与实现

我们试验了利用VR技术进行虚拟咬合仿真制作的全部过程。首先,采用光学三维测量仪对上下颌石膏模型进行数字化,通过预处理获取有效的三角网格曲面模型;其次,对咬合运动模型进行合理的简化,分解为一系列的平移运动和旋转运动;通过动态刷新完成开闭口运动、侧移运动的计算机运动仿真,可视化地观察咬合运动;然后利用模型碰撞检测算法动态地计算咬合接触位,并详细地分析了咬合接触时的咬合点位置分布和咬合剖切面上的咬合点接触关系;最后讨论了目前虚拟咬合仿真存在的问题和今后研究的方向。     

散发性急性戊型肝炎血清抗体动态变化和肝脏超微…

为探讨散发性戊型肝为血清抗体动态变化，应用酶联免疫法（ＥＩＡ）检测了７例急性戊型肝炎抗戊型肝炎病毒（ＨＥＶ）ＩｇＧ和ＩｇＭ抗体、并对１例２进行了肝超微结构病理检测，结果表明，发病１０天至４５天内抗ＨＥＶ－ＩｇＧ和ＩｇＭ滴度最高，发病第４０天仍有肝细胞肿胀，胞浆空化和线粒体固缩等病理变化。患者轿清抗－ＨＥＶＩｇＭ滴度在４５天后逐渐下降，２个月内全中消失，抗－ＨＥＶＩｇＭ滴度在４５天后逐渐下降，２个月     

The frequency of occurrence of autoantibodies against beta1-adrenoceptors and its clinical relevance in patients with hepatitis virus myocarditis

The aim of this study was to examine the frequency of occurrence of autoantibodies against beta1-adrenoceptors in patients with hepatitis virus myocarditis (HVM) and its possible correlation with clinical characteristics. A total of 103 patients with viral myocarditis were divided into a positive group (HVM group, n=29) and a negative group (Non-HVM group, n=74) according to the laboratory findings regarding their type of hepatitis virus. The study parameters included UCG, ECG, biochemical findings and screening of autoantibodies against beta1-adrenoceptor. It was shown that the positive rate of the hepatitis virus was 28.16% (29/103) in patients with viral myocarditis. The severity of myocardial or liver injuries and the frequency of occurrence of autoantibodies against beta1-adrenoceptors in patients with viral myocarditis were more pronounced, before treatment, in the HVM group than in the Non-HVM group. The positive rates of the antibodies against the hepatitis virus and the autoantibodies against beta1-adrenoceptors were highly consistent in patients with HVM (p<0.05). In conclusion, the frequency of occurrence of the autoantibodies against beta1-adrenoceptors may be one important marker of HVM and, thus, possibly involved in the pathogenesis of the HVM.     

人工肱骨头置换与肱骨近端锁定钢板治疗老年复杂肱骨近端骨折的近期临床疗效分析

[摘要] 目的 对比分析人工肱骨头置换术和肱骨近端锁定钢板治疗老年复杂肱骨近端骨折的近期疗效。方法 回顾性分析本院骨科2012年1月~2014年6月采用人工肱骨头置换术和肱骨近端锁定钢板治疗的老年（年龄>65岁）NeerⅢ、Ⅳ部分肱骨近端骨折患者54例，人工肱骨头置换治疗（置换组）30例，肱骨近端锁定钢板内固定治疗（内固定组）24例。比较分析两组手术时间、术中出血量、随访Neer评分及Constant-Murley评分来评定两组临床疗效。结果 两组随访时间相仿，平均随访时间为19.5个月（12~36个月）。内固定组手术时间（101±13）分钟，术中出血量（237.9±32.4）mL，Neer评分（82.5±3.2），Constant-Murley评分（71.7±5.0）；置换组手术时间（98±11）分钟，术中出血量（246.0±39.8）mL，Neer评分（82.9±4.5），Constant-Murley评分（73.5±5.0）。两组手术时间、术中出血量、随访Neer及Constant-Murley评分比较差异均无统计学意义（P＞0.05）。内固定组有1例出现肩关节半脱位，肱骨头置换组有2例出现肩关节半脱位，通过三角巾悬吊后关节脱位均得到纠正。有1例四部分骨折患者，行内固定术后出现内固定松动、肱骨头坏死，改行人工肱骨头置换术。结论 人工肱骨头置换术和肱骨近端锁定钢板治疗老年肱骨近端复杂骨折的近期临床疗效相似，对不可修复的肱骨近端骨折宜行肱骨头置换术。     

LuSIV cells: a reporter cell line for the detection and quantitation of a single cycle of HIV and SIV replication

A single cycle of viral replication is the time required for a virus to enter the host cell, replicate its genome, and produce infectious progeny virions. The primate lentiviruses, human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV), require on average 24 h to complete one cycle of replication. We have now developed and characterized a reporter assay system in CEMx174 cells for the quantitative measurement of HIV/SIV infection within a single replication cycle. The SIV(mac)239 LTR (-225 --> +149) was cloned upstream of the firefly luciferase reporter gene and this reporter plasmid is maintained in CEMx174 cells under stable selection. This cell line, designated LuSIV, is highly sensitive to infection by primary and laboratory strains of HIV/SIV, resulting in Tat-mediated expression of luciferase, which correlates with viral infectivity. Furthermore, manipulation of LuSIV cells for the detection of luciferase activity is easy to perform and requires a minimal amount of time as compared to current HIV/SIV detection systems. The LuSIV system is a powerful tool for the analysis of HIV/SIV infection that provides a unique assay system that can detect virus replication prior to 24 h and does not require virus to spread from cell to cell. Thus these cells can be used for the study of replication-deficient viruses and the high throughput screening of antivirals, or other inhibitors of infection.     

Identification and comparison of residues critical for cell-adhesion activities of two neutrophil CD66 antigens, CEACAM6 and CEACAM8

CEACAM6 (CD66c) and CEACAM8 (CD66b) are cell-adhesion proteins on neutrophils that belong to the human carcinoembryonic antigen (CEA) family. CEACAM6 reveals homophilic adhesion and heterophilic adhesion to other CEACAM family antigens including CEACAM8, CEACAM1, and CEA, whereas CEACAM8 exhibits only heterophilic adhesion to CEACAM6. Here, we investigated and compared structural requirements for the homophilic adhesion of CEACAM6 and heterophilic adhesion between CEACAM6 and CEACAM8 at the amino acid level by using CHO transfectants expressing their mutant and chimeric proteins. The NH(2)-terminal domain (N-domain) of CEACAM6 expressed on a CHO cell was suggested to bind the N-domain of CEACAM6 or CEACAM8 on the opposing cell. By homologue-scanning mutagenesis, we found that the locations of the sequences critical for the adhesion of CEACAM6 to itself and to CEACAM8 are overlapped and that they are highly similar but not identical to the locations of the residues previously shown to be essential for the binding of CEACAM antigens to Opa proteins of pathogenic NEISSERIAE: Our findings imply that subtle differences in the N-domain sequences determine the specificity of the CEACAM antigens on neutrophils for interaction with the same or different CEACAM antigens and the bacterial proteins.     

肺淋巴管平滑肌瘤病临床病理学观察

目的 探讨肺淋巴管平滑肌瘤病临床、病理特征。方法 对5例肺淋巴管平滑肌瘤病临床资料进行收集分析,HE切片观察,采用免疫组织化学(SP法)检测平滑肌肌动蛋白(SMA)、HMB45、基质金属蛋白酶(MMP)2、孕激素受体(PR)、雌激素受体(ER),并进行文献复习。结果 肺淋巴管平滑肌瘤病是原因不明的肺部疾病,只发生在女性,特别是绝经前妇女。临床表现为呼吸困难,咯血,气胸和乳糜胸等。病理学检查显示不同成熟度平滑肌细胞在细支气管壁、肺泡壁、淋巴管壁和血管壁周围增生,肺实质呈囊性变。增生的平滑肌细胞免疫组织化学5例SMA、HMB45、MMP2均阳性;1例的ER和PR均阳性,1例仅ER阳性,1例仅PR阳性,1例的ER和PR均阴性。结论 育龄期妇女如反复出现自发性气胸、咯血、活动后呼吸困难应考虑肺淋巴管平滑肌瘤病的可能,病理检查可确定肺淋巴管平滑肌瘤病的诊断。     

带臂外侧上皮神经及其营养血管筋膜皮瓣的应用解剖

目的:为带臂外侧上皮神经及其营养血管筋膜皮瓣提供解剖学基础.方法:32例经灌注红色乳胶的成人上肢标本,对臂外侧上皮神经及其营养血管等进行了较详细的应用解剖学研究.结果:臂外侧上皮神经在均由腑神经发出,起点横径为1.5±0.4mm,在三角肌深方斜向外下3.6±1.1cm从该肌后缘中1/3浅出肌间隔,分为上支和下支,分布于三角肌后部、外侧部和臂外侧上部.该神经的营养血管起源于旋肱后动脉,起点外径为0.9±0.4mm;其行程、分支和分布均同在神经,供血范因为14.8×9.8cm~2,并与周围的皮动脉存在丰富吻合.结论:带臂外侧上皮神经及其营养血管筋膜皮 瓣可视受区需要设计成游离瓣或旋转瓣,用于修复邻近部位、手或颌面部缺损.