自然感染途径建立肺炎链球菌感染性肺炎小鼠模型

目的探索自然感染途径制备肺炎链球菌感染性肺炎小鼠模型的方法,为肺炎链球菌性肺炎的相关研究提供实验基础。方法肺炎链球菌标准菌株(CMCC 31203)经血平板培养18 h后配成2麦氏浊度。小鼠浅麻醉状态下,破损鼻黏膜,滴注一定量肺炎链球菌菌液。于3、7、14和21 d分别处死小鼠,取肺脏进行组织病理学观察,确定肺炎小鼠模型制备的最佳方式。结果鼻黏膜破损组小鼠3 d时肺泡间隔内毛细血管扩张,肺泡腔内以红细胞和纤维素渗出为主;7 d时病变以纤维素和中性粒细胞浸润为主;14 d时肺泡腔内渗出减少,炎症开始减轻;21 d时肺脏外观趋于正常,肺泡腔内渗出物质基本溶解吸收。结论以3×107CFU的肺炎链球菌菌液通过破损的鼻黏膜感染7 d时可以制备出比较典型、稳定的肺炎小鼠模型。     

以乳突切迹和翼钩为基点的侧颅底分区方法

目的：探讨以乳突切迹和翼钩为基点的侧颅底分区新方法。方法：在乳突切迹后缘、翼钩、枕骨大孔前缘中点和颧根四个结构间相互连线,区分侧颅底并测量连线的长度。结果：乳突切迹与侧颅底重要结构的关系密切,切迹后缘与翼钩连线和正中线将侧颅底分成内、外侧两个大的三角区,每个区再分成前后两个三角区共四个三角区,即腭和颞下三角、咽三角、关节和听三角、血管神经三角,其中血管神经三角的三边长度左右侧分别为（74.52±5.47）mm和（74.66±5.41）mm、（59.77±3.84）mm和（59.67±3.56）mm、（42.23±3.11）mm和（42.48±2.60）mm。结论：本研究提供了新的侧颅底分区方法,且血管神经三角的区域划分更为科学,为临床侧颅底手术入路和定位提供了解剖学参考。     

小反刍兽疫研究概况

小反刍兽疫是严重危害山羊和绵羊等家畜的A类动物传染病,目前该病已经给我国养羊业造成了巨大的威胁。近年来,国内外学者对该病开展了许多研究工作,本文从病原学、流行病学、临床症状、发病机理、病理变化、诊断、防控措施等几个方面对该病研究概况进行了综述,为该病的有效防控提供有益的参考。     

不同加热方式对嗜肺军团菌环介导等温扩增检测法的影响

目的：研究不同的加热方式对嗜肺军团菌环介导等温扩增检测法的影响方法：用已知的13株嗜肺军团菌样本,采用空气浴、水浴和PCR仪同时进行环介导等温扩增,观察沉淀反应、荧光反应以及产物电泳结果。结果：水浴和PCR仪加热LAMP反应的沉淀产物较多,荧光反应较强,电泳检测结果较为明显。空气浴的3种检测结果均较弱。结论：采用水浴和PCR仪进行环介导等温扩增反应的效果较好,从仪器设备的成本及实验条件考虑,采用水浴是环介导等温扩增反应首选的加热方式。     

不同孔径CPC材料对大鼠骨髓间充质干细胞增殖的影响

目的：评价不同大小孔径的磷酸钙骨水泥（Calcium phosphate cement,CPC）材料对大鼠骨髓间充质干细胞（Bone mesenchymal stem cells,BMSCs）增殖能力的影响。方法：用盐析法制备三种不同孔径的（200-300μm、300-450μm、450-600μm）CPC材料,利用Micro-CT测量三种材料的平均孔径、孔隙率。无菌条件下取新生大鼠BMSCs原代培养并传代;将三组材料分别放置于24孔板内,每个材料接种5×104个细胞后,37℃、饱和湿度环境下静置培养。于接种后第1、4、7、14、21天用picogreen试剂盒测定细胞增值率;并在第14天、21天戊二醛固定材料并干燥喷金,扫描电镜观察材料表面细胞生长情况。结果：micro-CT测量结果显示：三种CPC材料孔径间相互连通,孔隙率均大于68%,平均孔径分别为235μm、422μm、505μm。细胞在三组材料上均呈对数增长趋势,在第14天到达平台期,在第1天三组细胞数量无明显差异,第4天450-600μm组细胞数量明显高于其余两组（P〈0.05）,在第7天细胞数量随孔径的增加而增加,3组间均有统计学差异（P〈0.05）,第14天和第21天200-300μm组细胞数量明显少于其余两组（P〈0.05）,300-450μm组和450-600μm组间无统计学差异（P〉0.05）。结论：孔径大小可影响大鼠BMSCs在多孔CPC材料上的增殖能力,随着孔径增大,细胞增殖力增高。本研究为进一步研究孔径结构对细胞的影响提供了实验依据。     

乳突切迹与侧颅底重要结构的解剖学观测及应用

目的:通过测量乳突切迹与侧颅底重要骨性结构的距离,为临床相关应用提供解剖学参考。方法:取成人颅骨标本50例(去颅盖标本8例,整颅42例)100侧,测量乳突切迹及其与侧颅底重要孔、裂和管的距离。结果:左右侧乳突切迹后缘距茎乳孔、颈静脉孔外侧缘、颈动脉管外口后缘、破裂孔、棘孔、卵圆孔距离分别为25.16±3.73cm和25.02±3.58cm、30.92±3.50cm和30.45±3.49cm、38.22±3.57cm和38.14±3.43cm、57.23±3.71cm和57.14±3.44cm、47.94±3.83cm和48.32±3.54cm、53.70±3.98cm和53.55±3.75cm。结论:以乳突切迹后缘做为侧颅底手术的定位标志能够为临床相关应用提供较方便、准确的定位参考。     

HIF-1α的表达在肝移植后肝癌复发与转移中的意义

目的:探讨肝癌组织中低氧诱导因子-1α(HIF-1α)表达与肝癌行肝移植治疗后肿瘤复发与转移之间的关系.方法:选择肝癌行全肝移植患者45例,应用免疫组化方法检测其切除的肝癌组织、癌旁组织中HIF-1α的表达,分析其表达与移植后肝癌复发与转移之间的关系和意义.结果:肝癌组织中HIF-1α阳性表达率明显高于癌旁组织(X<'2>=6.39,P<0.01).HIF-1α在肝癌组织中的表达与肿瘤直径、TNM分期、有无淋巴结转移、有无门静脉癌栓及肿瘤播散灶密切相关(P<0.01).复发转移组和未复发转移组HIF-1α阳性表达率比较,差异存在显著性(X<'2>=8.46,P<0.01).Cox回归多因素分析表明HIF-1α高表达是影响肝癌患者肝移植后肿瘤复发和转移的独立预后因素(P<0.01).结论:HIF-1α蛋白有望成为一个预测肝癌患者肝移植后复发和转移的重要指标.     

转化生长因子β1质粒对新鲜同种异体骨移植的免疫影响

目的:研究局部含注射转化生长因子β1(TGF-β1)基因的质粒对大鼠同种异体骨移植的免疫排斥反应的影响。方法:将64只Wister大鼠完全随机分成4组,每组16只,切取SD大鼠的胫骨移植到Wister大鼠人工建模后形成的胫骨缺损区。TGF-β1组于移植骨局部一次性注射含有TGF-β1基因的质粒(40μg/只);空质粒组于移植局部注射空质粒;免疫抑制剂组术前3天开始腹腔内注射环孢素A(10mg/kg)直至处死;异体移植组仅行异体骨移植,不予特殊处理。术后3、6、12周行光镜和电镜检查。结果:异体移植组和空质粒组骨基质排列紊乱,骨细胞消失,基质区内见块状低电子密度区。TGF-β1组和免疫抑制剂组可见骨细胞和完整的成骨细胞,骨小管状态良好。混合淋巴细胞培养(MLC)结果显示TGF-β1组(0.331±0.017)与免疫抑制剂组(0.501±0.004)差异无统计学意义(P>0.05),TGF-β1组与空质粒组(1.104±0.023)和异体移植组(1.206±0.019)差异有统计学意义(P<0.05)、结论:新鲜骨移植过程中局部注射TGF-β1质粒可发挥免疫抑制作用,降低宿主的免疫排斥反应。     

Uncovering the profile of somatic mtDNA mutations in Chinese colorectal cancer patients

In the past decade, a high incidence of somatic mitochondrial DNA (mtDNA) mutations has been observed, mostly based on a fraction of the molecule, in various cancerous tissues; nevertheless, some of them were queried due to problems in data quality. Obviously, without a comprehensive understanding of mtDNA mutational profile in the cancerous tissue of a specific patient, it is unlikely to disclose the genuine relationship between somatic mtDNA mutations and tumorigenesis. To achieve this objective, the most straightforward way is to directly compare the whole mtDNA genome variation among three tissues (namely, cancerous tissue, para-cancerous tissue, and distant normal tissue) from the same patient. Considering the fact that most of the previous studies on the role of mtDNA in colorectal tumor focused merely on the D-loop or partial segment of the molecule, in the current study we have collected three tissues (cancerous, para-cancerous and normal tissues) respectively recruited from 20 patients with colorectal tumor and completely sequenced the mitochondrial genome of each tissue. Our results reveal a relatively lower incidence of somatic mutations in these patients; intriguingly, all somatic mutations are in heteroplasmic status. Surprisingly, the observed somatic mutations are not restricted to cancer tissues, for the para-cancer tissues and distant normal tissues also harbor somatic mtDNA mutations with a lower frequency than cancerous tissues but higher than that observed in the general population. Our results suggest that somatic mtDNA mutations in cancerous tissues could not be simply explained as a consequence of tumorigenesis; meanwhile, the somatic mtDNA mutations in normal tissues might reflect an altered physiological environment in cancer patients.     

A novel synthesis of iminodiacetic acid: biocatalysis by whole Alcaligenes faecalis ZJB-09133 cells from iminodiacetonitrile

Iminodiacetic acid (IDA) has been widely used as an important intermediate in the fine chemical industry. In this study, a novel synthesis route of IDA from iminodiacetonitrile by whole microorganisms was investigated. A strain with the capability of producing nitrilase, ZJB-09133, was isolated and identified, and later named Alcaligenes faecalis ZJB-09133. In addition, the detailed biocatalysis of iminodiacetonitrile to produce IDA using ZJB-09133 was investigated. The results showed that the conversion reached 65.3% in Na(2)HPO(4)-NaH(2)PO(4) buffer of pH 8.0 under the following conditions: cells in the amount of 0.075-g DCW/L, 1.5% substrate, conversion time of 8 h, and a reaction temperature of 35°C. To the best of our knowledge, this is the first time that the production of IDA using a biocatalysis method has been reported.