Interspecies comparison of stellate cell‐containing macula flavae and vitamin A storage in vocal fold mucosa

The macula flavae (MF), populated by vitamin A‐storing stellate cells (SCs), are believed to play a fundamental role in development, maintenance and repair of the vocal fold (VF) mucosa; however, to date, they have mostly been examined in observational human cadaver studies. Here, we conducted an interspecies comparison of MF and SC phenotype, as well as vitamin A quantification and localization, in human, pig, dog, rabbit and rat VF mucosae. MF containing vitamin A‐positive SCs were only identified in human and rat specimens. Pig, dog and rabbit VF mucosae contained no discernable MF, but rather exhibited preferential vitamin A localization to mucous (pig), serous (dog) or mixed (rabbit) glands. This glandular vitamin A storage corresponded to exceedingly high concentrations of retinol in pig and dog mucosae, and retinyl ester in dog mucosa. These findings have significant implications for the presumed role of the MF and SCs in VF biology, the nature of vitamin A storage within the VF mucosa, and the selection of an appropriate animal model for future experimental studies.     

基于16S rDNA和COⅠ基因的3种血蜱分子生物学鉴定

目的利用16S rDNA基因及线粒体DNA细胞色素氧化酶亚基Ⅰ(cytochrome oxidase subunitⅠgene,COⅠ)基因建立3种形态学特征相近的血蜱属蜱类-长角血蜱、褐黄血蜱及铃头血蜱的分子生物学鉴定方法,探讨它们的系统发生关系。方法在上海市动物体表采集寄生蜱,在解剖显微镜下进行形态学鉴定后,提取蜱样本基因组DNA,采用PCR方法从蜱基因组中扩增16S rDNA及COⅠ基因,测序后进行同源性分析,用Mega 6.0软件分别构建系统发生树并进行系统进化分析。结果3种血蜱中长角血蜱的16S rDNA序列和COⅠ序列与GenBank中的长角血蜱的16S rDNA序列和COⅠ序列同源性分别为96.0%~97.2%,96.9%~98.8%;褐黄血蜱的16S rDNA序列和COⅠ序列与GenBank中的褐黄血蜱的16S rDNA序列和COⅠ序列同源性分别为95.9%~98.3%、87.1%~91.9%;铃头血蜱的16S rDNA序列与GenBank中来自日本的铃头血蜱(AB819170)的16S rDNA序列同源性为97.5%;3种血蜱16S rDNA基因之间的同源性分别为88.0%、87.0%、86.9%,COⅠ基因之间的同源性分别为80.4%、80.8%、82.8%。用16S rDNA及COⅠ基因构建系统发生树中,长角血蜱、褐黄血蜱分别与已知的长角血蜱和褐黄血蜱聚在一起,长角血蜱、褐黄血蜱均形成独立的分支。结论在传统形态学分类的基础上结合分子生物学鉴定方法能更准确地鉴定蜱的种类,并能更好地了解其系统发育进化关系。     

长角血蜱实验室繁殖规律研究与种群的建立

目的研究长角血蜱的繁殖规律和饲养条件,建立长角血蜱的实验种群。方法从自然界抓捕蜱,放在光照培养箱或干燥器中饲养,以小鼠和兔为饲血动物,对长角血蜱进行繁殖传代,记录饲血动物特异性、各虫态时间以及饲养条件。结果长角血蜱适宜养殖温度和相对湿度分别为（22±2）℃、（85±10）%,小鼠和兔可以作为长角血蜱的饲血动物,长脚血蜱一个生活周期约4～5个月。结论本研究基本摸清了长角血蜱的繁殖规律和饲养条件,并建立了实验种群。     

再次从福建分离出斑点热群立克次体

目的：从蜱类媒介中分离斑点热群立克次体。方法：用鸡胚卵黄囊培养法直接分离。结果：自福建省宁化县林区麂体表捉获的越原血蜱（Ｈａｅｍｅｐｈｙｓａｌｉｓｙｅｎｉ）中分离到１株斑点热群立克次体，经微量补结试验和ＰＣＲ检测初步确定为西伯利亚立克次体（Ｒｉｃｋｅｔｓｉａｓｉｂｉｒｉｃａ）。结论：进一步从病原学上证明了福建省存在北亚斑点热自然疫源地的可能性     

中国莱姆病螺旋体主要生物媒介的调查研究

为研究中国莱姆病螺旋体的生物媒介———蜱的种类、带菌状况及在传播莱姆病中的作用 ,采用布旗法和动物诱捕法在山林地区采集蜱 ,进行分类鉴定 ,应用直接荧光抗体染色法查蜱中肠带菌率 ,BSK培养基分离病原体 ,单克隆抗体鉴定菌株。调查蜱叮咬率与人群莱姆病感染率、患病率以及蜱的季节消长与莱姆病慢性游走性红斑 (ECM )病例发生的季节性关系。在全国 2 0个省 (市、区 )共采集到170 0 0多只蜱 ,分属为 2科 8属 2 3种 ,蜱类分布的地区差异较大 ,北方林区以全沟硬蜱为优势种 ,南方则以二棘血蜱和粒形硬蜱为优势种。 9种蜱的中肠携带莱姆病螺旋体 ,其带菌率以全沟硬蜱、二棘血蜱和粒形硬蜱为高 ,分别为 2 0 .0 0 %～ 4 5.0 0 %、16.0 0 %～ 4 0 .0 0 %和 2 4 .0 0 %。从 8种蜱分离出 10 8株莱姆病螺旋体 ,其单克隆抗体反应与美国B31菌株有明显的差异 ,绝大多数中国菌株对H6831呈阴性反应。蜱叮咬率高的人群 ,莱姆病的感染率和患病率也高。全沟硬蜱的季节消长与ECM发生的季节性相一致。证实多种蜱的中肠内携带莱姆病螺旋体 ;我国北方林区莱姆病螺旋体的主要生物媒介是全沟硬蜱 ,南方林区二棘血蜱和粒形硬蜱是重要的生物媒介。     

Transcriptional activities of two newly identified Haemaphysalis longicornis tick‐derived promoter regions in the Ixodes scapularis tick cell line (ISE6)

Ticks are obligate haematophagous ectoparasites considered to be second to mosquitoes as vectors of human diseases and the most important vector for animals. Despite efforts to control tick infestations, they remain a serious health problem. Gene manipulation has been established in mosquitoes and led to the control of mosquito populations and of mosquito‐borne pathogens. Therefore, gene manipulation could be useful for controlling ticks and tick‐borne pathogens. To investigate effective gene expression vectors for ticks, the promoter activities of commercial plasmids were evaluated in a tick cell line (ISE6). Dual luciferase assays revealed that pmirGLO, the human phosphoglycerate kinase promoter contained plasmid vector, showed the highest activity in ISE6 cells amongst the tested plasmids. Moreover, we identified the promoter regions of the Haemaphysalis longicornis actin (HlAct) and the intracellular ferritin (HlFer1) genes. To construct a more effective expression vector for ticks, these promoter regions were inserted into pmirGLO (pmirGLO‐HlAct pro and pmirGLO‐HlFer1 pro). The pmirGLO‐HlAct pro vector showed significantly higher promoter activity than pmirGLO, whereas the pmirGLO‐HlFer1 pro vector demonstrated significantly lower promoter activity than pmirGLO in ISE6 cells. The HlAct promoter region may have high promoter activity in ISE6 cells. The results of the present study provide useful information for the development of a genetic modification system in ticks.     

日本血蜱的染色体

本文报告日本血蜱的核型:2n=22(20 XX)和2n=21(20 X)。少数胚细胞另有1B染色体,即2n=20 XX B。性别决定属XX-XO遗传系统。     

Differentiation potential of the cells in the macula flava of the human vocal fold mucosa

The latest research suggests cells in the maculae flavae located at both ends of the lamina propria of the human vocal fold mucosa have stemness. This study investigated the differentiation potential of the cells in the maculae flavae of the human vocal fold mucosa. Four normal human adult vocal folds from surgical specimens were used. After extraction of the anterior maculae flavae located at the anterior end of the lamina propria of the human vocal fold mucosa under microscope, the maculae flavae were minced, cultured and proliferated in mesenchymal stem cell growth medium and morphological features were assessed. Cell surface markers were detected using flow cytometry. Cell differentiation into adipogenic, chondrogenic and osteogenic lineages was performed. Cell’s differentiation potential was assessed using a human pluripotent stem cell functional identification kit and immunohistochemistry. Subcultured cells formed a colony-forming unit. Subcultured cells expressed CD90, CD105 and CD73 and lacked expression of CD45, CD34, CD11b, CD19 and HLA-DR. They differentiated into adipogenic, chondrogenic and osteogenic lineages. Consequently, the cell features in the maculae flavae meet the minimal criteria defining mesenchymal stromal cells. In addition, subcultured cells differentiated into ectoderm, mesoderm and endoderm and expressed stage-specific embryonic antigen 3 (SSEA-3). The results of this study are consistent with the hypothesis that the cells in the maculae flavae in the lamina propria of the human vocal fold mucosa are putative stem cells.     

胸椎黄韧带骨化症的手术疗效分析

目的探讨胸椎黄韧带骨化症的手术治疗方法及其影响因素。方法回顾性分析我院1998-2005年采用后路整块半关节突全椎板切除减压术治疗的76例病人中已随访到的40例OTLF患者的资料,对手术疗效进行随访,应用秩检验进行统计学分析,判断患者年龄、术前病程、JOA评分与病变节段数与手术疗效的关系。结果获得随访的40例病人,平均随访时间56个月,优15人,占37.5%;良17例,占42.5%;可5例,占12.5%;差3例,占7.5%。优良率80%,有效率92.5%。JOA评分与病变节段数对术后疗效有显著影响。结论后路整块半关节突全椎板切除减压术是行之有效的手术方式。脊髓损伤程度轻、病变节段少者术后疗效好。     

孤雌生殖长角血蜱各虫期形态的扫描电镜观察

用扫描电镜对采自四川地区孤雌生殖的长角血蜱的各期虫体及虫卵进行了形态学观察。结果:雌成蜱的口下板齿式4︱4或4—5︱5—4,多孔区小孔30～35个,两多孔区的间距为孔区长径的2倍,气门板“D”字状,肛毛5对;若蜱齿式2—3︱3—2,须肢第3节背面后缘无刺,无多孔区和生殖孔,气门板亚圆形,无气门斑,肛毛3对;幼蜱齿式2︱2,无气门板,肛毛1对,无肛后沟,爪垫明显超过爪长的2/3。各期虫体的哈氏器结构基本相同,但成蜱前窝内感毛7根,而若蜱、幼蜱均为6根。卵表面光滑无特殊结构。首次观察到孤雌生殖长角血蜱成蜱气门板腹面前方有7、8个小孔,肛瓣下方有密集成带的小刺以及十几个气孔样物。