脑微血管形态参数和细胞化学定量参数测量法

本文介绍采用图象分析仪测量脑微血管一些形态定量参数和细胞化学定量参数的方法。脑毛细血管的一些三维形态参数根据数学形态学和体视学原理进行测量和计算。这些定量参数对于微血管和微循环的定量研究和临床诊断有一定的实用价值。     

肝癌组织中线粒体DNA D-Loop区碱基变异与ROS水平

为了探讨ROS水平与突变的关系,对原发性肝癌线粒体DNA区的突变情况进行研究,同时对原发性肝癌患者组织细胞内ROS进行测定。选择20例原发性肝癌组织及其邻近的癌旁组织,用PCR方法将线粒体DNA D-Loop扩增后测序。组织内ROS的水平采用流式细胞技术测定。结果表明在20对原发性肝癌组织中存在8对mtDNA突变,突变率为40%,共发现突变位点53个,包括2个插入,11个缺失,40个点突变,其中T-C,C-T的转换占75%,4个属于微卫星结构。癌组织突变一般伴有癌旁组织突变,癌组织突变位点高于癌旁组织。发现一例标本的癌组织和癌旁组织均有大片段丢失。原发性肝癌组织内ROS水平明显高于癌旁对照( P<0.01),同时我们发现在区发生突变的患者的组织中ROS水平明显高于未发生突变的肝癌组织标本（P<0.01）,发生突变的癌旁组织内ROS水平明显高于未发生突变的癌旁组织（P<0.01）。结论 (1)线粒体DNA D-Loop区是一个高度多态性和突变性的区域,在原发性肝癌中突变率较高。(2)肝癌患者组织细胞内ROS异常,提示肝癌的线粒体DNA发生的点突变及肝癌的发生可能与ROS升高有关。     

华南长江和西江流域鲤科宽鳍鱲和马口鱼不同线粒体DNA谱系之间的体形分化

本文研究了两种鲤科鱼类,即分布于华南长江与西江的宽鳍鱲和马口鱼的不同线粒体DNA谱系之间的体形差异。近期基于线粒体DNA的这两个物种的系统发育分析认为这两个类元内包含多个谱系(宽鳍鱲A-D和马口鱼1-5)。本文采用几何形态度量学的方法研究了这些不同线粒体DNA谱系间是否存在体形方面的差异。在宽鳍鱲的4个不同的线粒体DNA谱系中,有3个被纳入本研究内容。结果显示:在宽鳍鱲的3个线粒体DNA谱系中,2个具有体形的差异。在马口鱼的5个线粒体DNA谱系中,3个被纳入本文的研究。结果显示所有3个谱系均存在体形差异。因此,线粒体DNA的谱系差异很大程度上表现在谱系间的体形差异方面,表明线粒体DNA谱系可能对应于不同的物种。     

肝细胞癌及相关病变的计算机图像定量分析

为研究肝炎、肝硬变与肝细胞癌的关系,利用自动图像分析仪对６４例肝炎、肝硬变、癌旁肝硬变、肝细胞癌和正常肝组织进行了十三项参数的形态定量研究。结果：大多数参数随病变的发展而呈规律性变化,癌旁肝硬变的多数参数介于不伴肝癌的肝硬变与肝细胞癌之间。提示：（１）慢活肝、肝硬变与肝细胞癌密切相关;（２）癌旁肝硬变不同于不伴肝癌的肝硬变,与肝细胞癌的关系更密切。利用逐步判别分析选择出六项参数建立判别函数方程,回代正确率为９８．２％。     

华南长江和西江流域鲤科宽鳍鳞和马口鱼不同线粒体DNA谱系之间的体形分化

本文研究了两种鲤科鱼类,即分布于华南长江与西江的宽鳍雠和马口鱼的不同线粒体DNA谱系之间的体形差异。近期基于线粒体DNA的这两个物种的系统发育分析认为这两个类元内包含多个谱系（宽鳍鱲A—D和马口鱼1—5）。本文采用几何形态度量学的方法研究了这些不同线粒体DNA谱系间是否存在体形方面的差异。在宽鳍馘的4个不同的线粒体DNA谱系中,有3个被纳入本研究内容。结果显示：在宽鳍雠的3个线粒体DNA谱系中,2个具有体形的差异。在马口鱼的5个线粒体DNA谱系中,3个被纳入本文的研究。结果显示所有3个谱系均存在体形差异。因此,线粒体DNA的谱系差异很大程度上表现在谱系间的体形差异方面,表明线粒体DNA谱系可能对应于不同的物种。     

口腔鳞状细胞癌线粒体DNA的HVRⅢ区突变的研究（英文）

目的：检测口腔鳞状细胞癌患者线粒体DNA复制控制区（mtDNA D-loop）高变Ⅲ区（hypervariable regionⅢ,HVRⅢ）的突变情况,并探讨其意义。方法：以口腔鳞状细胞癌患者癌旁组织及正常组织作为对照,对7例口腔鳞状细胞癌组织样本的mtDNA D-loop HVRⅢ区进行PCR扩增和测序分析。结果：在7例患者的癌组织、癌旁组织、正常组织样本中共发现72个（56种）核苷酸改变,其中51个（26种）为核苷酸多态性改变;3个肿瘤组织样本中共发现21个突变,其中16个位于HVRⅢ区范围内;癌旁组织及正常组织未发现突变;口腔鳞状细胞癌的mtDNA D-loop HVRⅢ区突变率为42.9%（3/7）。结论：mtDNA D-loop HVRⅢ区的变异可能与口腔鳞状细胞癌的易感性有一定的联系;本研究为寻找新的肿瘤基因诊断和肿瘤遗传易感性的标志物提供了依据。     

微囊藻毒素-LR 对小鼠肝细胞线粒体功能的影响*

摘要目的：探究微囊藻毒素-LR 对小鼠肝细胞线粒体功能的影响。方法：采用BALB/c 小鼠作为模型动物,随机分为3 组：A 组, 空白对照组,正常饮用水;B 组,添加5 g/L微囊藻毒素-LR 的饮用水;C 组,添加30 g/L 微囊藻毒素-LR 的饮用水。分组喂养3 个月,分离小鼠肝脏、提取线粒体,采用线粒体荧光探针JC-1 测定线粒体膜电位（MMP）,qRT-PCR检测自噬相关基因Beclin1 和 Lc3琢的转录水平,Western Blot检测细胞色素C的释放,电镜观察线粒体的形态和内部结构。结果：微囊藻毒素-LR 处理组的小 鼠肝细胞线粒体膜电位明显下降,自噬相关基因Lc3琢的转录水平上升,细胞色素C由线粒体释放到胞浆,电镜观察线粒体形态 异常、内部结构被破坏。结论：微囊藻毒素-LR 对小鼠肝细胞线粒体有较强的毒性作用,并引发线粒体自噬。     

微囊藻毒素-LR对小鼠肝细胞线粒体功能的影响

目的：探究微囊藻毒素-LR对小鼠肝细胞线粒体功能的影响。方法：采用BALB／c小鼠作为模型动物,随机分为3组：A组,空白对照组,正常饮用水;B组,添加5g门L微囊藻毒素．LR的饮用水;C组,添加30gm微囊藻毒素-LR的饮用水。分组喂养3个月,分离小鼠肝脏、提取线粒体,采用线粒体荧光探针JC-1测定线粒体膜电位（MMP）,qRT-PCR检测自噬相关基因Beclinl和Lc3α的转录水平,WesternBlot检测细胞色素c的释放,电镜观察线粒体的形态和内部结构。结果：微囊藻毒素-LR处理组的小鼠肝细胞线粒体膜电位明显下降,自噬相关基因Lc3α的转录水平上升,细胞色素C由线粒体释放到胞浆,电镜观察线粒体形态异常、内部结构被破坏。结论：微囊藻毒素-LR对小鼠肝细胞线粒体有较强的毒性作用,并引发线粒体自噬。     

《实用生物体视学技术》

由申洪、沈忠英编著的《实用生物体视学技术》已由中山大学出版社出版。体视学是由二维结构信息定量推论三维结构信息的一门新兴边缘学科。该书从实用性出发,阐述了体视学的基本原理,着重介绍了体视学的基本测试方法,并就体视学中的样本、抽样,数据处理等作了详细介绍。此外还介绍了图像分析系统及其在生物医学中的应用和平面图像常用定量指标及相应计算公式等。本书不仅包括体视学经典方法,而且包括体视学的新概念、新参数、新方法、新进展及作者的一些经验,体会和研究成果;注重理论与应用结合,注重实际工作中的具体问题,对具体操作和测算方法有举例说明,图文并茂,深入浅出。     

白花蛇舌草对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响

目的:探讨白花蛇舌草(注射液)对人胃癌细胞MNK-45线粒体膜电位及凋亡相关基因表达的影响。方法:将人胃癌细胞MNK-45分成4组,每组设置3个复孔,对照组为未加入白花蛇舌草的MNK-45细胞,3组实验组分别加入终浓度为20、30、40μg/ml白花蛇舌草的MNK-45细胞,各组在5%的CO₂培养箱中孵育48 h后,利用激光共聚焦显微镜下观察细胞形态变化,流式细胞术检测线粒体膜电位,qRT-PCR检测Cytochrome C (Cyt c)、caspase3和caspase9基因的表达变化,Western blot检测Cytochrome C (Cyt c)、caspase3和caspase9蛋白的表达变化。结果:与对照组相比,终浓度为20、30、40μg/ml的各白花蛇舌草处理组,其MNK-45细胞的线粒体膜电位均明显降低(P<0.01),Cyt c、caspase3和caspase9的基因表达均明显上调(P<0.01)、蛋白表达也均显著升高(P<0.05或P<0.01),40μg/ml的白花蛇舌草处理组的表现最佳。结论:在终浓度为20～...     

线粒体融合素基因-2在正常体重和肥胖乳腺癌患者不同组织中表达的研究

目的研究线粒体融合素基因(mfn2)在正常体重和肥胖乳腺癌患者的不同组织中的表达情况,探讨mfn2基因与乳腺癌发生的关系及评价肥胖与乳腺癌发病的关系。方法采用RT-PCR技术对20例正常体重乳腺癌患者及20例肥胖乳腺癌患者的胸大肌组织,脂肪组织,正常乳腺组织,癌旁组织和癌组织的mfn2的表达水平进行了测定和对比分析。结果两组乳腺癌患者不同组织的mfn2的表达水平有差异。在正常体重组内,胸大肌组织和正常乳腺组织内的表达无显著性差异(P=0.101),但显著高于脂肪组织(P=0.016),高于癌旁组织(P=0.027),高于癌组织(P=0.002)。肥胖组内,胸大肌组织、脂肪组织、正常乳腺组织内的表达无显著性差异(P>0.05),但显著高于癌旁组织(P=0.003),高于癌组织(P=0.001)。两组间比较,mfn2在肥胖组患者的五种组织中的表达较正常体重组均有显著性降低(P<0.05)。结论mfn2基因在癌组织中低表达,且在肥胖患者组织中表达更低,mfn2与乳腺癌的发生有关且肥胖可能增加患乳腺癌的风险。     

c-myc oncogene gene dosage, serum CEA and CA-15.3 antigen levels, and cellular DNA values in relation to ex vivo chemosensitivity of primary human breast cancer

A pilot study on relationships of selected molecular factors (c-myc oncogene average gene copy numbers (AGCN); serum CEA and CA 15.3 antigen levels; tumor cells' DNA values), to the ex vivo chemosensitivity of primary female human breast cancer in a modified adenosine triphosphate cell viability chemosensitivity assay (ATP-CVA), was performed. Four drug combinations were tested. A group of 75 cases of female primary breast cancer was assessed. Numerous correlations were found among molecular factors tested but none, with the exception of tumor grading, of these reflected ex vivo chemosensitivity of tumors tested. The results suggest that the parameters tested may not be important factors related to adjuvant chemoresponsiveness of primary human breast cancer to tested drug combinations.     

Evolution of sexual preferences in quantitative characters

An analysis of equilibria and dynamics of the means, variances, and covariances of female mating preference for a quantitative male secondary sexual character following a Gaussian model is presented. For many combinations of viability and sexual selection parameters the evolving Gaussian distribution of phenotypes can diverge. The results on the cases of convergence and their limiting forms suggest some reinterpretations of Fisher's "runaway" process of sexual selection. One possibility is to interpret Fisher's postulated "initial advantage not due to female preference" as a shift in viability selection where runaway evolution occurs if the mean preferred trait evolves beyond its new viability optimum (due to sexual selection). This definition is contrasted with situations in which the new viability optimum is undershot. The quantitative and qualitative conclusions differ from models that approximate genetic covariance evolution involving a constant covariance.     

Estimating carrying capacity in a newly colonized sand fly Lutzomyia serrana (Diptera: Psychodidae)

The phlebotomine sand fly Lutzomyia serrana (Damasceno & Arouck) was mass-reared tinder conditions of varying densities in an effort to improve colony production efficiency. To do this, the experimental carrying capacity of a standard rearing chamber was determined, i.e., the optimum population size in relation to density (individuals per unit of space). Rearing chambers of 100 cm3 were populated with 1-50 L. serrana engorged females and an equal number of males. Laboratory conditions were maintained at 23-26 degrees C and 85-95% RH. The following parameters were recorded for each experimental chamber (three replicates): (1) female mortality without oviposition, (2) number of eggs oviposited and (3) number of adults emerging from the egg cohort. Female mortality began to increase substantially in the 26-female chamber, from 5.7% to 15% and finally reaching 60.2% in the 46-50 female chambers. In the chambers containing 1-20 females, egg number and realized adult progeny increased linearly to reach an asymptote. In the 20-50 female chambers, the number of eggs ranged from 420 to 699, and adult production from 306 to 432. The optimum carrying capacity for the 100-cm3 chambers was 22 +/- 2 females. Beyond this number, auto-regulation was initiated, i.e., female mortality without oviposition increased as the number of females per chamber increased. Total number of eggs and adult production was similar in all chambers containing 20-50 females. In conclusion, for optimizing production of mass reared sand flies, determination of the carrying capacity is essential to optimize use of insectary resources, to avoid loss of valuable potentially ovipositing females, and to increase overall production efficiency.     

Sampling strategies for using female gametophytes to estimate heterozygosity in conifers

Summary Unbiased estimators of genotype and allele frequencies and their respective variances are obtained for loci identified by mendelian segregation in haploid female gametophytes from individual trees. By a minimum sampling variance criterion, the allocation of experimental effort between the number of female gametophytes analysed per tree and the number of trees sampled per population is examined for a fixed total amount of experimental effort. For estimating heterozygosity, the optimum sampling design for many (generally most) cases is three female gametophytes per tree, but may be more than three depending upon the true genotype frequencies in the population. For estimating allele frequencies, the optimum sampling design is one female gametophyte per tree except in cases where a strong negative correlation exists between alleles within genotpyes. Guidelines are discussed for determining a suitable number of female gametophytes to be analysed per tree in order to estimate heterozygosity.     

常规超声及超声造影在肝癌中的诊断及鉴别诊断价值

目的:探讨常规超声及超声造影在肝癌中的诊断及鉴别诊断价值。方法:收集2009年5月至2013年10月在我院进行住院诊治的肝癌患者120例,选择飞利浦IU22进行常规超声检查,选择飞利浦的IU22进行超声造影检查。结果:常规超声检查68例为低回声,20例为强回声,12例为等回声,17例为混合性回声,3例误诊为肝外病变,检出率为97.5%。肝癌组织的AT、TTP、ACT值明显晚于正常肝组织(P0.05),而BI、PI、MTT与BF值对比差异无明显差异(P0.05)。Spearman相关分析显示超声造影灌注参数AT、TTP、ACT与肿瘤分化程度呈正向相关性(P0.05)。结论:常规超声及超声造影在肝癌中的诊断及鉴别都有很好的应用价值,超声造影的应用可反映正常和病变组织的血流灌注情况,从而提高肝癌诊断的准确性。     

Estimation of sojourn time in chronic disease screening without data on interval cases

Estimation of the sojourn time on the preclinical detectable period in disease screening or transition rates for the natural history of chronic disease usually rely on interval cases (diagnosed between screens). However, to ascertain such cases might be difficult in developing countries due to incomplete registration systems and difficulties in follow-up. To overcome this problem, we propose three Markov models to estimate parameters without using interval cases. A three-state Markov model, a five-state Markov model related to regional lymph node spread, and a five-state Markov model pertaining to tumor size are applied to data on breast cancer screening in female relatives of breast cancer cases in Taiwan. Results based on a three-state Markov model give mean sojourn time (MST) 1.90 (95% CI: 1.18-4.86) years for this high-risk group. Validation of these models on the basis of data on breast cancer screening in the age groups 50-59 and 60-69 years from the Swedish Two-County Trial shows the estimates from a three-state Markov model that does not use interval cases are very close to those from previous Markov models taking interval cancers into account. For the five-state Markov model, a reparameterized procedure using auxiliary information on clinically detected cancers is performed to estimate relevant parameters. A good fit of internal and external validation demonstrates the feasibility of using these models to estimate parameters that have previously required interval cancers. This method can be applied to other screening data in which there are no data on interval cases.     

Sexual antagonism leads to a mosaic of X-autosome conflict

Males and females have different optimal values for some traits, such as body size. When the same genes control these traits in both sexes, selection pushes in opposite directions in males and females. Alleles at autosomal loci spend equal amounts of time in males and females, suggesting that the sexually antagonistic selective forces may approximately balance between the opposing optima. Frank and Crespi noted that alleles on the X chromosome spend twice as much time in diploid females as in haploid males. That distinction between the sexes may tend to favor X-linked genes that push more strongly toward the female optimum than the male optimum. The female bias of X-linked genes opposes the intermediate optimum of autosomal genes, potentially creating a difference between the direction of selection on traits favored by X chromosomes and autosomes. Patten has recently argued that explicit genetic assumptions about dominance and the relative magnitude of allelic effects may lead X-linked genes to favor the male rather than the female optimum, contradicting Frank and Crespi. This article combines the insights of those prior analyses into a new, more general theory. We find some parameter combinations for X-linked loci that favor a female bias and other parameter combinations that favor a male bias. We conclude that the X likely contains a mosaic pattern of loci that differ with autosomes over sexually antagonistic traits. The overall tendency for a female or male bias on the X depends on prior assumptions about the distribution of key parameters across X-linked loci. Those parameters include the dominance coefficient and the way in which ploidy influences the magnitude of allelic effects.     

Stromal myofibroblasts in breast cancer: relations between their occurrence, tumor grade and expression of some tumour markers

It is suggested that tumour stromal myofibroblasts exert an unfavourable effect on the biology of breast cancer. We are aware of only a single study which examined relationships between manifestation of myofibroblasts in the stroma of breast cancer and clinicopathological data of the patients. The present study was aimed at estimation of the effect exerted by myofibroblasts present in the tumour stroma on principal pathological parameters and on expression of Ki67, P53 and HER-2 proteins in the group of the most frequent breast cancers, the ductal cancers. In paraffin sections of 60 ductal breast cancers (20 cases in G1, 20 in G2 and 20 in G3), immunohistochemical reactions were performed to detect expression of smooth muscle actin (SMA) in order to visualize myofibroblasts, Ki67, P53 and HER-2. The studies demonstrated that the most numerous myofibroblasts were present in G3 cases and they were the least frequent in G1 cases (P = 0.02). Positive correlations were observed between the presence of myofibroblasts in tumour stroma and expression of Ki67 and HER-2 in breast cancer cells in the entire group (P < 0.001 and P = 0.001, respectively), in G2 cases (P = 0.003 and P = 0.03) and in G3 cases (P = 0.01 and P = 0.03). Considering that the higher grade, Ki67 and HER-2 are thought to represent unfavourable prognostic factors, the elevated content of myofibroblasts in tumour stroma is probably typical for cases with worse prognosis.     

四氧嘧啶诱发糖尿病模型效果的性别差异

目的了解性别因素对四氧嘧啶诱发糖尿病动物模型的影响,为提高动物模型的复制效率提供实验依据。方法分别给雌、雄比格犬和昆明小鼠注射不同剂量的四氧嘧啶,药后3、7、14、21 d测定血糖值,同时统计实验期间动物的死亡情况。结果给予同等剂量的四氧嘧啶,雌性比雄性动物的血糖升高更快,浓度更高。雌性犬四氧嘧啶的最适造模剂量为40 mg/kg,而雄性犬在此剂量下的模型成功率只有40%,二者差异极显著（70%VS40%,P〈0.01）;雄性犬的最适使用剂量为50 mg/kg,但在此剂量下有高达30%的雌性犬因高血糖而死亡。四氧嘧啶对小鼠的影响与犬基本一致,雌雄鼠的最佳剂量分别为200 mg/kg和250 mg/kg。结论雌性动物对四氧嘧啶的敏感性较雄性动物高,雄性动物在使用四氧嘧啶复制糖尿病模型时,其剂量通常需要较雌性动物高20%左右。