一腓骨肌萎缩症家系临床及基因分析

目的分析一腓骨肌萎缩症家系的临床表现及不同基因检测方法的特点。方法收集一CMT家系8名成员临床资料,并应用等位基因特异性PCR-双酶切方法及多重连接依赖的探针扩增技术(MLPA)检测PMP22基因突变情况,同时选择60名性别、年龄无明显差异的健康人做为对照组。结果该家系中患病者以行走不稳、跨阈步态,伴有弓形足为主要临床表现。该家系中5名成员经等位基因特异性PCR-双酶切及MLPA方法均检测出PMP22基因重复序列,其中出现临床症状的有4名(Ⅱ3、Ⅱ9、Ⅱ11、Ⅲ7),未出现临床症状但基因检测结果示PMP22基因重复序列的为携带者有1名(Ⅲ5),家系中余3名成员及对照组60名均未见重复序列。结论基因检测在明确CMT诊断中起重要作用,且MLPA法筛查基因时操作更简便、灵敏度更高、特异性更好。     

光化学作用对肿瘤细胞胞吞大分子的释放

目的观察不同光敏剂在肿瘤细胞内的分布,以及光化学作用对肿瘤细胞胞吞大分子的释放作用.方法使用激光共聚焦显微镜观察ALA、ALA-HE、HMME、TPPSa2在肿瘤细胞SW480、K562的胞内分布,使用FITC-右旋糖酐观察肿瘤细胞对荧光标记大分子的胞吞作用,通过光照激发光敏化细胞,动态观察光化学作用前后胞吞大分子FITC-右旋糖酐的胞内分布变化.结果ALA、ALA-HE、HMME、TPPSa24种光敏剂均表现为胞质分布,核内分布少.ALA、ALA-HE的胞内分布主要表现为胞质内的弥散性荧光.HMME则表现为胞质内颗粒状荧光与弥散荧光同时存在.TPPSa2为典型的胞质内颗粒状荧光分布.光敏剂在SW480、K562两种肿瘤细胞的胞内分布没有明显差异.光照激发不同光敏剂对肿瘤细胞胞吞FITC-左旋糖酐的胞内分布影响不同.由TPPSa2及HMME活化而产生的光化学作用表现出明显的荧光颗粒重分布,ALA、ALA-HE则对胞吞荧光无明显重分布作用.结论不同光敏剂因其不同的理化性质而表现为不同的胞内分布.光化学作用可对肿瘤细胞胞吞大分子产生胞吞释放作用,其作用机制可能与光化学作用对胞吞泡的破坏有关.     

C端带Myc和多聚组氨酸双重标签基因的改构体J-HNP-1在COS-7细胞的转染表达

目的改构α-防御素-1(HNP-1)使其成为一端带J链的改构体J-HNP-1分子,并探索建立能有效表达和分泌J-HNP-1,其表达产物又易于被检测和分离纯化的哺乳动物细胞表达系统。方法利用重组PCR技术,使HNP-1一端带上J链;将J-HNP-1 DNA片段插入哺乳动物细胞表达载体pcDNA3.1(-)/Myc-HisC中,构建出C端带Myc和6×His的rpcDNA3.1(-)/Myc-HisC/J-HNP-1;将此重组真核表达载体导入COS-7细胞,分别从mRNA和蛋白质水平分析J-HNP-1的表达情况,并同步检测细胞可溶性蛋白及培养上清的抗菌活性。结果采用RT-PCR法从被转染的细胞中扩增出1条约786bp的片段,其大小与预测相符;采用Western印迹法,用抗His抗体检测到细胞可溶性蛋白及培养上清在相对分子质量约24×103处有强反应条带显示;细胞可溶性蛋白及培养上清的抗菌活性检测显示,经rpcDNA3.1(-)/Myc-HisC/J-HNP-1转染的COS-7细胞的可溶性蛋白和培养上清均有明显的抗菌活性。结论α-防御素-1(HNP-1)被成功改构成杀菌肽J-HNP-1;所建立的J-HNP-1哺乳动物细胞表达载体系统能有效表达和分泌活性J-HNP-1。     

Characterizing luminous efficiency functions for a simulated mesopic night driving task based on reaction time

The objective of this study is to test the luminous efficiency functions V(lambda), V'(lambda), V(10)(lambda) and their linear combinations on the basis of a data set gained from a simulated mesopic night-time driving experiment. Another aim is to provide 'real-world' data for the 'X framework' or 'linear combination model', and to find out its limits in a practical situation. Human performance was measured by the reaction time method. Results show that the single parameter of the linear combination of photopic and scotopic luminous efficiency functions can be determined analytically with little variation for a given mesopic background luminance level and a given visual target colour, but the computation leads to considerable deviations comparing all three target colours (red, green and blue) used in the experiment. The conclusion for the given experimental conditions is that the single parameter of the linear combination model has an increasing deviation for lower background luminance levels.     

Analysis of the strain relatedness of oral Candida albicans in patients with diabetes mellitus using polymerase chain reaction‐fingerprinting

To increase our understanding of Candida pathogenicity, the identification of those strains most frequently associated with infections is of paramount importance. Polymerase chain reaction (PCR)‐based methods are extremely effective in differentiating and determining reproducibility, they require minimum starting material and are rapid and simple to perform. In this study, the genetic relatedness of Candida albicans was assessed for two geographically different patient groups (London, UK and Parma, Italy) affected by diabetes mellitus. C. albicans samples from the oral cavities of non‐diabetic healthy subjects were also examined by PCR fingerprinting to evaluate the possible genetic differences among endogenous strains in individuals with and without diabetes mellitus. PCR fingerprinting, with subsequent phylogenetic analysis of C. albicans isolates from the diabetic patients from London and Italy and from the non‐diabetic subjects, revealed that there were significant differences (P < 0.0001) between C. albicans isolates indicative of the distinct ecological niches that occur in the oral cavities of these patient cohorts. The most diverse group comprised the isolates from the diabetic patients in the UK, possibly reflecting the antifungal treatment that these patients had received. Further studies that include isolates from patient cohorts with systemic diseases other than diabetes mellitus, and from more diverse geographic localities are required to explain the relatedness of C. albicans isolates in the mouth.     

An experimentally supported, mathematical explanation of the gas chromatographic elution behaviour of the long-chain carbon members of the homologous series

By substituting the saturated vapour phase tension of the pure normal hydrocarbons described by the Clausius-Clapeyron law into the accepted expression of the specific retention volume (V_g,T), a theoretically coherent and relatively simple mathematical evidence of the elution behaviour of the homologous members has been deduced. It gives exponential retention time dependence on carbon number for isothermal, and nearly equidistant (i.e., approximately linear retention-time dependence on carbon number) elution for linear temperature programmed gas chromatographic runs. The final equations are in close correlation with the experimental results. Special emphasis is placed on the fact that a good approximation—not strict physical laws—have been found.     

中药膳食配合运动干预对肥胖性脂肪肝的影响

目的 探讨中药膳食配合运动干预对肥胖性脂肪肝的影响。方法 将68例脂肪肝患者随机分为对照组和观察组各34例。对照组常规给予低脂饮食和易善复、大黄庶虫丸等降酶保肝治疗；观察组按常规方法治疗外，采用中医辨证施食予以活血化淤、健脾利湿的中药膳食疗法配合运动干预．疗程均为1年。结果 治疗后观察组体重、甘油三酯、总胆固醇、谷丙转氨酶等显著低于对照组（均P〈0．01）。结论 中药膳食配合运动干预对肥胖性脂肪肝疗效显著。     

间期细胞原位PCR检测B细胞系恶性淋巴瘤

目的：应用原位PCR技术检测B细胞系恶性淋巴瘤。方法：采用免疫球蛋白重链（IgH)基因第三互补决定区（CDR－Ⅲ）的引物，dig-11-dUTP标记物，应用直接原位PCR技术检测Ⅲ-Ⅳ级恶性淋巴瘤的间期淋巴细胞，结果：在初诊的15例恶性淋巴瘤患中，12例检测到IgH基因重排；4例临床表现呈现淋巴瘤性白血病患，治疗后骨髓完全缓解，2例间期细胞原位PCR仍显示有IgH基因重排，6个月复发，结论：间期细胞原位PCR技术能敏感、准确地检测Ⅲ级以上的恶性淋巴瘤，并帮助预测复发，为一有效的基因诊断方法。     

静脉毒瘾者84例HGV感染状况

目的调查庚型肝炎病毒（ＨＧＶ）在静脉毒瘾者中的感染状况。方法采用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）检测８４例静脉毒瘾者血浆标本。ＨＧＶＲＮＡ经热变性法提取后逆转录为ｃＤＮＡ，在ＨＧＶ５′非编码区（５′ＮＣＲ）设计两对引物进行巢式扩增，产物为２３８ｂｐ，并经限制性内切酶ＨｐａⅡ鉴定扩增产物来自ＨＧＶ。结果８４例中有１５例为ＨＧＶＲＮＡ阳性，阳性率为１７．９％。ＨＧＶＲＮＡ阳性病例中１１例合并丙型肝炎病毒感染（１１／１５）。结论静脉毒瘾者是ＨＧＶ感染的高危人群；不洁注射是获得ＨＧＶ感染的重要途径。