有效中药复方研究的方法学探讨

从化学的角度探讨了临床有效中药复方研究的四个过程 ,简述了直接试验设计在全方药味筛选及化学模式识别中药质量控制中的应用 ,论述了复方有效部位和有效部位中主要有效成分的确定及化学计量学的应用和主要药效物质化学结构信息的计算机模拟方法 ,为中药复方研究提供一个范例。     

Protection of chickens against highly lethal H5N1 and H7N1 avian influenza viruses with a recombinant fowlpox virus co-expressing H5 haemagglutinin and N1 neuraminidase genes

Inactivated whole avian influenza virus (AIV) vaccine provides protection against homologous haemagglutinin (HA) subtype virus, but poor protection against a heterologous HA virus. Moreover, it induces chickens to produce antibodies to cross-reactive antigens, especially nucleoprotein, which is limits AIV serological surveillance. In this study, a recombinant fowlpox virus co-expressing HA (H5 subtype) and NA (N1 subtype) genes of AIV was evaluated for its ability to protect chickens against intramuscular challenge with a lethal dose of highly pathogenic (HP) AIV. Susceptible chickens were also vaccinated by wing-web puncture with the parent fowlpox vaccine virus. Following challenge 4 weeks later with HPAIV, all chickens vaccinated with recombinant virus were protected, while the chickens vaccinated with either the unaltered parent fowlpox vaccine virus or unvaccinated controls experienced 100% mortality following challenge. This protection was accompanied by the high levels of specific antibody to the respective components of the recombinant vaccine. The above results showed that rFPV-HA-NA could be a potential vaccine to replace current inactivated vaccines for preventing AI.     

125~I－后标记检测DNA－蛋白质交联物的研究

１２５~Ｉ－后标记法是庄志雄等人（１９９４）提出的一种检测ＤＮＡ－蛋白质交联物（ＤＰＣｓ）的新技术，该法在检测离体ＣＨＯ细胞ＤＰＣｓ的实验中已显示出敏感、快速和简便的优点。本研究表明，１２５~Ｉ－后标记法不仅能有效地检出由紫外光和铬酸钾诱导的ＣＨＬ细胞ＤＰＣｓ，并能有效而敏感地检测由铬酸钾和氯化镍引起的大鼠不同组织特别是白细胞ＤＰＣｓ形成的情况，是一种敏感的和可考虑应用于人群调查的ＤＰＣｓ检测方法。     

白癜风患者血小板的研究

测定７５例白癜风患者周围血象、血小板相关抗体（ＰＡＩｇＧ）、血小板粘附及聚集、第３因子功能、高岭土部分凝血活酶时间（ＫＰＴＴ）、一期凝血酶原时间（ＰＴ）。结果发现白癜风患者血小板减少，ＰＡＩｇＧ增高、聚集时间延长。提示这种异常与免疫有关。     

酵母硒对小鼠EAC瘤生长和抗氧化机能的影响

目的观察酵母硒对小鼠艾氏腹水瘤(EAC)生长及机体氧化机能的影响。方法将30只小鼠分为3组(对照组、低剂量酵母组和高剂量酵母组),酵母硒组小鼠通过灌胃的方法分别灌注酵母硒溶液,剂量分别为107μg/kg.BW.d和214μg/kg.BW.d,灌胃第13 d接种EAC,第23 d后测定小鼠肿瘤重和血清中丙二醛(M DA)含量、谷胱甘肽过氧化酶(G SH-Px)和超氧化物歧化酶(SOD)活力。结果酵母硒可抑制小鼠EAC的生长,同时显著提高血清G SH-Px活性(P<0.05),极显著提高血清SOD活力(P<0.01),高剂量酵母硒降低血清M DA水平(P<0.05)。结论酵母硒可抑制小鼠移植肿瘤的生长,显著提高机体抗氧化能力。     

JC病毒样颗粒可直接转运进入细胞核

目的探讨JC病毒(JCV)病毒样颗粒(VLP)是否可以直接转运进入细胞核。方法心用JCV主要外壳蛋白VP1体外表达、重组VIP，在其表面标记异硫氰基荧光素(FTTC)，同时在其内部包裹荧光染料Cy3，感染培养的HeLa细胞和SVG细胞，荧光显微镜观察VLP入核转运。结果HeLa和SVG细胞感染包裹Cy3的FTTC-VLP时，FTTC与Cy3同时出现于细胞核内相同部位；而感染FTTC—VP1与Cy3混合物时，FTTC虽可在细胞核内检测到，但Cy3信号几乎消失。包裹Cy3的VIP用SDSPAGE展开，荧光显像后行考马斯亮蓝(CBB)染色，发现Cy3和VLP移行至不同部位，证明Cy3不能与VP1结合，提示VLP以完整的颗粒形式转运进入细胞核。应用包裹外源性DNA的VLP感染培养的HeLa和SVG细胞，发现包裹的DNA在细胞浆和细胞核内均可检测到，提示JCV入核过程与VIP相同。结论VLP可以不经裂解直接转运进入细胞核，JCV入核转运可能与VLP相同。     

Cytokine expression and endothelial cell and lymphocyte activation in human cardiac allograft rejection: an immunohistochemical study of endomyocardial biopsy samples.

We used monoclonal antibodies and immunohistochemical staining of frozen tissue sections to study the expression of cytokines in human cardiac allograft rejection. The 113 endomyocardial biopsy samples were stained for interleukin (IL)-2, IL-6, and interferon-gamma. The findings were compared to expression of the endothelial cell adhesion molecule ICAM-1, and the lymphocyte receptor for the adhesion molecule VCAM-1, VLA-4. Four biopsy samples from patients with idiopathic cardiomyopathy served as controls. IL-2 was not expressed in lymphocytes of controls and only occasionally in mild or moderate cellular rejection, humoral rejection, and Quilty lesions. IL-2 expression was prominent in severe cellular rejection. Interferon-gamma expression increased in proportion to the severity of cellular rejection and was not expressed in other conditions. IL-6 staining, which was only observed in occasional cases, was mild. Cytokine and adhesion molecule expression tended to increase with the severity of cellular rejection. This study shows that cytokine expression can be documented in human allograft endomyocardial biopsy samples with immunohistochemical techniques. The findings support the concept of an important role for cytokines in human cardiac allograft rejection.     

PCR-SSCP法检测结核分枝杆菌耐药性

目的：探讨耐多药结核分枝杆菌耐药基因突变与耐药性的关系以及聚合酶链反应-单链构象多态性分析(poly merase chain reaction-single strand cinfomlation polymorphism，PCR-SSCP)方法的临床应用价值。方法：用PCR-SSCP方法检测58株结核分枝杆菌临床分离株katG，rpoB，rpsL基因突变并与常规药敏试验检测结果进行对比。结果：经常规药敏试验检测，58株结核分枝杆菌临床分离株中共有41株耐药，其中，耐异烟肼(INH)为35株，高耐药株27株；耐利福平(RFP)为31株，高耐药株24株；耐链霉素(SM)有31株，高耐药株26株。同时耐3种药物的有21株(51．2％)，耐2种药物的14株(34．1％)，单耐药株6株(14．6％)．PCR-SSCP方法对58株临床分离株katG，rpoB，rpsL基因突变的检测率为40％(23／58)，45％(26／58)，38％(22／58)，其中检出3个基因同时突变的有13株(32％)，2种基因突变的12株(29％)，1种基因突变的有10株(23．8％)．常规药敏试验与PCR-SSCP法检出结核分枝杆菌同时耐3种药物的符合率为61．9％(13／21)，检出耐2种药物的符合率为85．70k，(12／14)，检出耐1种药物的符合率为50％(3／6)．高耐药株中突变率为80．5％(62／77)，低耐药株中突变率为60％(12／20)．结论：PCR-SS-CP方法对耐2种以上药物的结核杆菌检出率较高，且耐药基因突变率随着耐药浓度增高而增高。将PCR-SSCP法与药敏试验联合应用可互相弥补，已成为临床指导用药的好方法。     

一株产新型大环内酯抗生素海洋解淀粉芽孢杆菌的分离鉴定

Objective To perform taxonomic studies on a novel macrolactin-producing gram-positive bacterial strain MS1721 isolated from the mud at a depth of 30 m in the East China Sea. Methods Taxonomic studies were conducted based on the phenotypic and BIOLOG analysis, 16S rDNA gene sequencing and blast analysis, and fatty acid components analysis. The antibacterial activity of macrolactin S was determined by evaluation models established in our lab. Results MS1721 strain displayed high similarity with genus Bacillus in similarity values between 97%～99%, in which the nearest neighbor was Bacillus amyloliquefaciens with the similarity value of 99.6%. The predominant fatty acid component was 15:1 iso H/13:0 3OH. Conclusions MS1721 strain belongs to the genus Bacillus, and Bacillus amyloliquefaciens MS1721 was proposed as the name for this strain on the basis of the taxonomic data. The second metabolite-macrolactin s produced by this strain exhibited obvious antibacterial activity.     

腹腔镜下处理输卵管积水对体外受精胚胎移植结局的影响

目的探讨腹腔镜下处理输卵管积水对体外受精胚胎移植(in vitro fertilization and embryo transfer, IVF-ET)结局的影响.方法 2002～2005年在我院行IVF-ET伴输卵管积水的53例分3个研究组:①未处理组17例,仅行IVF-ET,未对积水进行处理;②直接切除组17例,行IVF-ET术前在腹腔镜下处理输卵管积水;③失败后切除组19例,IVF-ET失败后腹腔镜下处理输卵管积水后再行IVF-ET.结果未处理组新鲜移植周期妊娠率15.8%(3/19),解冻周期妊娠率10.5%(2/19),平均周期妊娠率13.2%(5/38),累积活胎率17.6%(3/17);直接切除组分别为36.8%(7/19),23.1%(3/13),30.3%(10/32),41.7%(8/17);失败后切除组分别为16.7%(2/12),58.3%(14/24),44.4%(16/36),73.7%(14/19).失败后切除组的解冻周期妊娠率、平均周期妊娠率及累积活胎率均高于未处理组(χ2＝10.374,P＝0.001;χ2＝8.903,P＝0.003;χ2＝11.305,P＝0.001),直接切除组平均周期妊娠率及累积活胎率呈高于未处理组的趋势(χ2＝3.377,P＝0.066;χ2＝3.360,P＝0.067).结论输卵管积水降低IVF-ET的妊娠率,妊娠结局不良,腹腔镜下处理输卵管积水后再行IVF-ET提高妊娠率,获得良好的妊娠结局.