染色体原位抑制杂交鉴别染色体易位的研究

本文报告作者运用荧光标记的６号、９号、１３号、１５号、和Ｙ染色体探针，对１２名患者的Ｇ显带外周血、骨髓或胎儿羊水细胞染色体标本进行染色体原位抑制杂交的研究。     

荧光原位杂交技术及染色体核型分析在产前诊断中的应用

目的：探讨荧光原位杂交技术（FISH）及染色体核型分析技术在产前诊断中的应用价值。方法应用FISH 技术对未培养的羊水间期细胞进行13、16、18、21、X／Y 号染色体数目检测，同时对培养后的羊水中期细胞进行染色体核型分析。结果600例羊水标本 FISH 检测均得出结果，检测成功率为100％，FISH 检测出31例阳性结果，异常核型检出率为5．2％。600例羊水标本染色体核型分析中598例检测出结果，2例标本污染无法分析，检测成功率为99．7％。598例检测结果中阳性结果63例，异常核型检出率为10．5％。结论 FISH 检测羊水胎儿染色体非整倍体的方法过程简便、快速，成功率高，结论可靠。但只能检出特定染色体的非整倍体，应用有一定的局限性。与传统染色体核型分析技术相互联合、补充，可以更好地应用于产前诊断中。     

应用荧光原位杂交技术进行产前诊断的可行性探讨

目的：探讨荧光原位杂交技术（FISH）应用于绒毛及羊水间期细胞进行产前诊断的可行性。方法 应用18、X号染色体着丝粒探针及21号染色体特定区域探针,分别对绒毛及羊水间期细胞进行荧光原位杂交,并与中期分裂相进行比较分析。结果 绒毛及羊水间期细胞均出现了杂交信号,万以绒毛细胞杂交比率高,信号强;常染色体探针出现3个及1个杂交点的比率约为1％和6％,X染色体探针出现3点的比例也在1％左右。结论 荧光原位杂交技术对检出染色体非整倍体有较强的特异性,同时也为染色体非整倍体的检出提供了诊断依据。FISH技术应用于产前诊断具有快速、简便、灵敏的优点,有较大的临床应用价值。     

2830例孕中期羊水细胞染色体核型分析

目的 分析妊娠中期进行产前诊断的高危孕妇羊水细胞染色体核型,了解此期异常核型出现的频率、类型及与各种产前诊断指标的关系。方法2006年1月-2009年7月来我中心就诊的2830例有产前诊断指征的孕妇,妊娠17～24W,在B超定位下行羊膜腔穿刺术,抽取羊水细胞进行培养及染色体核型分析。结果在2830例羊水检查的标本中,发现染色体多态性110例,占3．89％,主要为1qh＋、inv（9）和Y染色体的异染色质区域的变异;发现异常核型102例。占3．60％。平衡易位为主要的染色体异常（23例）,占异常核型的22．5％（23／102）其次为非整倍体,其中,21三体占23．5％（24／102）,18三体次之,占13．7％（14／102）,其他的异常核型如罗伯逊易位、部分缺失、衍生染色体等,所占比例较小。结论通过对广东地区产前诊断高危孕妇的羊水,在有产前诊断指征的孕妇,胎儿染色体异常的发生率为3．60％,染色体平衡易位、三体、尤其21三体是妊娠中主要的异常核型,是高龄最常见的染色体异常。     

1550例遗传咨询及细胞遗传学分析

目的 总结和探讨1550例遗传咨询者中染色体异常的发生情况。方法 抽取受检者外周血，常规微量法培养及制备染色体标本，G显带运动核型分析。结果 在1550例受检查中共检出染色体异常87例，异常率为5．61％。其中有不良孕产史者833例中，染色体异常34例；不孕不育症109例中，染色体异常8例；闭经及性腺发育不良102例中，染色体异常19例，智能发育不全者172例中，染色体异常24例，其它334例中，染色体异常2例。结论 染色体异常与许多生育及发育缺陷有关，对这些患者进行染色体检查以明确诊断很有必要。     

无培养羊水细胞染色体荧光原位杂交法初探

荧光原位杂交法 (ＦＩＳＨ)是一种快速、准确的染色体测定方法。为了引进ＦＩＳＨ技术用于产前诊断 ,我们抽取了中期妊娠引产妇女的羊水标本 ,用ＦＩＳＨ法对无培养羊水细胞间期的性染色体进行了检测。材料与方法一、标本来源 :16例中期妊娠引产病例 ,年龄 19～ 30岁 ,平均年龄 2 5岁。孕周 17～ 31周 ,平均 2 2孕周。全部病例无心、肝、肺、肾及内分泌疾病史 ,采用雷凡诺尔羊膜腔内注射引产术 ,在注入雷凡诺尔前先抽取羊水 ,若抽出羊水为血性或黄绿色 (胎粪污染 ) ,则此病例被排除。 16例全部抽取羊水 5ｍｌ,色澄清淡黄色 ,送实验室处理。…     

荧光原位杂交技术产前诊断常见染色体非整倍体

目的:探讨荧光原位杂交(HSH)技术产前诊断常见染色体非整倍体的应用价值.方法:采用13、18、21、X和Y染色体特异性DNA探针对108例孕20～25周孕妇的未培养羊水细胞进行HSH检测,同时行羊水细胞染色体核型分析.结果:HSH检测108例成功104例(96.3%),其中染色体数目正常100例(58例为46,XX;42例为46,XY),染色体数目异常4例(3例47,XX,+21;1例45,X/46,XX).正常染色体数目中99例与传统核型分析结果一致,而另1例HSH结果为46,XX者传统核型分析显示为47,XX,+mar.染色体数目异常的4例与传统核型分析结果完全一致.核型分析为47,XX,+mar者因标记染色体来源不是13、18、21、X和Y染色体故HSH未能诊断.对于13、18、21、X和Y特异性探针来说,敏感性、特异性及与核型分析的符合率均为100%.结论:HSH技术用于产前诊断常见染色体非整倍体,具有简便、快速、特异性强、敏感性高、所用样本量少等优点.     

两种FISH试剂盒检测羊水染色体的比较

目的应用两种荧光原位杂交(FISH)试剂盒检测羊水染色体,比较两者在临床的应用效果。方法采用北京金菩嘉和美国雅培Vysis生产的FISH试剂盒对30例未培养的羊水进行染色体检测,比较两者在信号强度和杂交率的差异。结果 30例样本均获得FISH检测结果,且两种试剂临床诊断符合率达到100%,杂交率无统计学意义(P>0.05)。Vysis试剂21/13染色体荧光杂交信号较强,金菩嘉试剂18染色体信号清楚,不易融合。结论 Vysis与金菩嘉两种FISH检测试剂应用染色体异常产前诊断均可达到同样的临床诊断效果。     

22q11.2微缺失在贵州少数民族先天性心脏病患者中的研究

目的 探索22q1i.2微缺失在贵州少数民族先天性心脏病患者中的发病情况.方法 选取44例苗族、侗族及布依族先天性心脏病患者,在UCSC数据库中选择合适的BAC克隆,自制双色BAC探针,采用双色细菌人工染色体-荧光原位杂交技术对上述患者进行22q11.2微缺失检测.结果 在2例法洛四联症患者中检测到22q11.2微缺失,缺失率为4.5％(2/44).结论 在苗族、布依族、侗族等少数民族先天性心脏病患者中存在22q11.2微缺失现象,对发现的阳性患者应密切观察其行为和认知方面的发育情况,一旦发现异常,应予以积极干预.     

妊娠丢失夫妇染色体核型分析的临床意义

目的探讨染色体异常与妊娠丢失的关系。方法收集260对妊娠丢失夫妇的临床资料,采用传统的细胞遗传学方法进行外周血淋巴细胞培养和染色体核型分析。结果48例标本染色体异常,发生率为18．5％（48／260）,包括28例（58．3％）结构异常、18例（37．5％）多态性改变、2例（4．2％）数目异常。染色体结构异常主要是平衡易位（71．4％,20／28）。偶发性与复发性妊娠丢失（RPL）夫妇之间的染色体异常发生率差异无统计学意义（P〉0．05）。结论染色体异常可导致妊娠丢失,平衡易位是其主要原因,夫妇双方孕前进行染色体检查有助于优生指导。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.