广东汉族人群补体C2 , Bf, C4的遗传多态现象

对110例广东汉族人血清作了补体C2, Bf, C4的测定,其基因频率分V1为：C2*C'0.9500, C2*B: 0.0227,C2-,4:0182, C2*QO:O．0091;Bf*S：0．8364, Bf^`F:0．1409, Bf*S07：0．0091, Bf *S025： 0.009i,Bf*S055：0,0045; C4*A3：0.6327,C4*A4：0．1327,C4*_00：0．1020, C4*A5：0．0255 （一4*A2: 0·0918,C4*,41：0．0053;C4*B1：0．4569, C4*B2：0．4416, C4*QO:O．0558,C4*B5：0．0152,C4"}B96： 0.0152, C4*B3:0.0102, C4*B92:0.0051。木调查在我国首次发现一例C2*QO纯合子。     

中国武汉地区汉族人补体第四成份(C4)的遗传多态现象

对神经氨酸酶处理去涎的血浆进行高压琼脂糖电泳后,分别应用免疫固定和溶血覆盖技术调查了我国武汉地区汉族180人的C4多态现象。在C4A座位发现5个变型:C4A4、3、2、1和91;在C4B座位发现6个变型:C4B 3、2、1、92、9W和96。两个座位均有静息等位基因(C4A~*QO和C4B~*QO)存在。它们相应的基因频率为:C4A~*4,0.014;3,0.633;2,0.192;1,0.011;91,0.003;QO,0.147;C4B~*3,0.006;2,0.127;1,0.751;92,0.041;9W,0.003;96,0.006;QO,0.0660单零C4A(c4A QO)表型个体占总体28.3％,单零C4B(C4B QO)占11.1％;纯合C4A缺乏(C4A QO,QO)和纯合C4B缺乏(C4BQO,QO)分别占0.56％。和1.11％卡方测验表明,C4A和C4B基因频率分别符合Hardy-Weinberg遗传平衡定律。     

第13届国际生物奥林匹克竞赛(2002)实验试题答案

(续 2 0 0 3年第 38卷第 6期第 5 7页 )实验 动物系统学和形态学Q1.　 (5分 )A B C D E F G H I J0 1 0 1 0 2 0 4 0 1 0 1 0 1 0 1 0 5 0 3Q2　 (5分 )A B C D E F G H I J0 1 0 2 0 4 0 5 0 1 0 6 0 3 0 70 80 9Q3　 (10分 )A B C D E F G H I JK + + +L +M ++ +N +O +P +R ++S ++T ++ +++ +Q4 A　 (4分 ) 　□　□　　 　□　□Q4 B　 (1分 )　　□Q5　 (5分 )A B C D E F G H I J0 3 0 1 0 2 0 2 0 3 0 3 0 1 0 1 0 3 0 2Q6　 (10分 )　　 A.Gammaruspulex(钩虾 )　 B.Anopheles sp.(疟　蚊 )　 C.H irudo …     

烟叶果胶质分解菌的选育

从烟叶叶面分离到产果胶酶真菌 18株 ,在此基础上 ,进行液体培养 ,测定酶活 ,得到一株酶活性较高的黄曲霉DPE - 0 0 5。对该菌株产酶培养基的碳、氮源进行正交法研究 ,正交实验的结果表明 ,影响该菌产酶活性的因素依次为A(麸皮 ) >B(乳糖 ) >C(果胶 ) >D(硫酸铵 ) ,其最佳组合为A2B3C3D1。最适产酶条件为 :麸皮 4 0 % ,果胶 0 3% ,乳糖 1 5 % ,(NH4 ) 2 SO4 0 5 % ,KH2 PO4 0 2 5 % ,MgSO4 ·7H2 O 0 0 5 % ,NaNO30 0 2 % ,FeSO4 ·7H2 O 0 0 0 1% ,起始pH 6 0 ,在 2 8℃摇床培养 7d产酶量达到最高。以玉溪B3F烟叶为材料 ,施加DPE - 0 0 5菌株所产酶液 ,在 5 0℃贮存 12h。经化学成分检测 ,结果表明 ,果胶质降低了 18 15 %。     

第12届国际生物奥林匹克竞赛(2001)理论试题(3)

(续 2 0 0 2年第 7期第 6 1页 )理论试题 A答案1.D　 2 .D　 3.B　 4 .D　 5 .C　 6 .C　 7.D　 8.A　 9.C10 .C　 11.A　 12 .D　 13.B　 14 .D　 15 .删除　 16 .D17.C　 18.B　 19.B　 2 0 .A　 2 1.A　 2 2 .D　 2 3.D2 4 .B　 2 5 .C　 2 6 .B　 2 7.D　 2 8.D　 2 9.A　 30 .D31.删除　 32 .D　 33.C　 34.B　 35 .D　 36 .C　 37.A38.删除　 39.删除　 4 0 .C　 4 1.A　 4 2 .D　 4 3.C4 4.C　 4 5 .D　 4 6 .A　 4 7.C　 4 8.C　 4 9.D　 5 0 .B5 1.删除　 5 2 .C　 5 3.A　 5 4.D　 5 5 .B　 5 6 .C　 5 7.A…     

双侧自体腘绳肌腱一期重建前后交叉韧带损伤的临床疗效

目的：探讨关节镜辅助下使用双侧自体腘绳肌腱一期修复膝关节前后交叉韧带损伤的方法和临床疗效。方法：内窥镜微创双侧自体腘绳肌腱修复膝关节内韧带,术后用IKDC分级、影像学IKDC分级、Lysholm功能评分和KT2000TM测量进行关节机能打分。结果：11例患者获得3—5年随访,平均随访3．8年。术前Lysholm功能评分平均（46．8±5．7）分,终末随访时平均（81．3±10．5）分,差异有显著性（P〈0．05）。术后关节稳定性测量,在20磅时、30磅和最大拉力时健膝和患膝分别是：6．1±0．3和6．8±0．8;6．3±0．5和7．7±1。3;7．5±0．6和9．6±2．4,统计学上差异无显著性（P〉0．05）。主观IKDC分级：A级4例,B级6例,C级1例;影像学IKDC分级：A级8例,B级2例,C级1例。结论：关节镜辅助下使用双侧自体腘绳肌腱一期修复膝关节前后交叉韧带损伤是重建膝关节稳定性的良好有效方法。     

复合诱变选育高效转化DHEA为7α,15α-diOH-DHEA的菌株及其转化工艺优化

结合酮康唑抗性筛选法,采用亚硝基胍和低能氮离子注入复合诱变方法筛选得到一株高效生物转化去氢表雄酮(D H E A)为3β,7α,1 5α-三羟基雄甾-5-烯-1 7-酮(7α,1 5α-d i O H-D H E A)的菌株亚麻刺盘孢C o l l e t o t r i c h u m l i n i S T-1,该突变株在底物D H E A投料浓度为1 0 g/L时产物摩尔得率达到3 4.2%,较出发菌株提高了4 6.2%。在此基础上进行培养基组分的优化,采用P l a c k e t t-B u r m a n实验设计考察转化培养基中各组分对产物摩尔得率的影响,有效筛选出葡萄糖、酵母粉和M g S O4·7 H2O浓度对产物摩尔得率影响显著,继而采用最陡爬坡路径逼近最大响应区域,并利用中心组合响应面设计实验对3个显著性因素的最佳水平进行研究,得到最适转化培养基组分为(g/L):葡萄糖2 6.3 4;酵母粉1 2.1 5;玉米浆3.0 0;F e S O4·7 H2O 0.0 1 5;M g S O4·7 H2O0.1 4;K H2P O40.9 0。采用该优化培养基,菌株C.l i n i S T-1的产物摩尔得率达到4 9.3%,较优化前提高了4 4.2%。     

乳杆菌制剂(定君生)和(或)甲硝唑三种方法治疗滴虫性阴道炎疗效比较

目的探讨滴虫性阴道炎的更佳治疗方法。方法2004年10月至2005年9月绍兴市妇幼保健院妇科门诊对92例滴虫性阴道炎病例随机分为A、B、C三组,A组31例,采用乳杆菌制剂0．25g塞阴道,每晚1次,共10d;B组31例．采用甲硝唑片0．2g口服,3次／d,共7d,同时用乳杆菌制剂塞阴道;C组30例,采用甲硝唑栓0．5g塞阴道,每晚1次,共10d,同时用甲硝唑片口服。结果停药3～5d的治愈率：A组67．74％、B组93．55％、C组93．33％;A组与B、C组之间差异有显著性（P〈0．05）,B组与C组间差异无显著性（P〉0．05）。停药3月的治愈率：A组41．93％、B组87．10％、C组56．67％,B组与A、C组间差异有非常显著性（P〈0．01）,有效率：A组54．84％、B组93．55％、C组70．00％,B组与A、C组间差异有显著性（P〈0．05）,A组与C组间治愈率及总有效率差异无显著性（P〈0．05）。结论甲硝唑片口服联合乳杆菌制剂（定君生）塞阴道是一种治疗滴虫性阴道炎的更佳方法,值得临床推广应用。     

慢性低O2高CO2对大鼠脑组织的损伤作用及川芎嗪对其影响的研究

目的：探讨慢性低O2高CO2条件下脑烯醇化酶（NSE）、蛋白S100、超微结构等改变及川芎嗪对其影响。方法：将SD大鼠分为正常对照组（A组）,低O2高CO2组（B组）,低O2高CO2＋川芎嗪组（C组）,采用免疫组织化学、透射电镜方法,观察平均肺动脉压（mPAP）、颈动脉压（mCAP)、血清NO浓度、脑NSE、S100变化及脑超微结构等改变。结果：①B组mPAP明显高于A组,C组低于B组,三组间mCAP无明显差异。②B组血清NO浓度低于A组（P＜0．01）,C组血清NO浓度高于B组（P＜0．01）。③B组比A组NSE、S100吸光度值降低,C组比B组NSE、S100吸光度值增高。④B组神经元、神经胶质细胞水肿,C组神经细胞损害明显减轻。结论：慢性低O2高CO2脑NSE、S100降低,川芎嗪对脑损害有保护作用。     

神经刺激仪经肌间沟定位臂丛神经分支行臂丛麻醉效果分析

目的：比较神经刺激仪经肌间沟定位臂丛神经分支行肌间沟臂丛神经阻滞麻醉的效果及安全性。方法：选择拟行肌间沟臂丛神经阻滞的上肢手术的患者80例,ASAI或II级,随机均分为A组和B组,每组40例患者。两组均给予1％的利多卡因＋0．375％耐乐品20mL。记录完成操作所需时间、感觉神经阻滞起效时间、感觉神经阻滞完善时间、运动神经阻滞起效时间、运动神经阻滞完善时间;评价手术区域麻醉效果（优、良、差、失败）;观察并记录并发症。结果：A组完成操作所需时间（5．01±1．40）min,明显长于B组（2．83＋O．87）min（P〈0．01）。A组感觉阻滞起效时间（4．48±1．36）min,明显短于B组（7．0±2．06）min（P〈0．01）;A组运动阻滞起效时间（4．88＋±1．18）min,明显短于B组（7．0±1．67）min（P〈0．01）。A组感觉阻滞完善时间（11．73±3．62）短于B组（13．33±3．02）min（P=0．033）。A组运动阻滞完善时间（11．18±2．73）短于B组（12．41±2．48）min（P=0．038）;麻醉效果优等率A组为87．5％,B组为67．5％,差异有统计学意义x^2=4．588,P=0．032;优良率A组为97．5％,B组为90．0％,差异无统计学意义x2=1．920,P=0．166;A组、B组均未出现严重麻醉并发症。结论：A组行肌间沟臂丛神经阻滞比B组阻滞操作时间长,但神经阻滞麻醉效果好,神经阻滞完善率高。     

Alzheimer病患者APP及PS1基因表达量的研究

为了检测Alzheimer病（Alzheimer’s disease,AD）患者外周血中淀粉样前体蛋白（Amyloid Precursor Protein, APP）基因及早老素1（Presenilin 1, PS1）基因的表达情况,进而探讨APP及PS1基因的表达与AD的相关性,采用SYBRGreenⅠ的方法对45例AD患者、25例血管性痴呆（vascular dementia, VD）患者及60名正常对照组样本的mRNA进行绝对定量,检测得到APP基因及PS1基因在对照组中的表达水平分别为0.026±0.005 amol/μg cDNA和0.026±0.004 amol/μg cDNA;在AD患者组中的表达量分别为0.044±0.006 amol/μg cDNA和0.051±0.011 amol/μg cDNA;,在VD患者组中的表达水平分别为0.072±0.013 amol/μg cDNA和0.039±0.005 amol/μg cDNA 。经显著性检验,AD患者组APP基因的表达水平上调,t=2.639, P<0.01;PS1基因的表达水平同样呈上调趋势,t=2.173,P<0.05,差异均具有统计学意义。VD患者组APP基因的表达水平上调,t=3.028,P<0.01;PS1基因的表达水平也同样呈上调趋势,t=2.012,P<0.05,均有显著性差异。因此,APP及PS1基因的表达水平的增高并不一定与AD发生特异性关联,而可能与多种导致痴呆的脑部病变发生关联。     

Time-Dependent Increase of Chitinase1 in APP/PS1 Double Transgenic Mice

It is reported that chitinase1 increases in Alzheimer’s disease (AD). However, the alteration of chitinase1 in the progress of AD is still unclear. Thus, we designed the present study to detect chitinase1 level in different stages of APP/PS1 double transgenic mice. Experimental models were APP/PS1 double transgenic mice with 4, 12 and 22 months. Cognitive function was detected by Morris water maze test in APP/PS1 mice as well as controls. ELISA and the quantitative RT-PCR were used to detect chitinase1 level in different groups. The study displayed that expression of chitinase1 gradually increased in a time-dependent manner in APP/PS1 mice, while there were no statistical differences among the wild-type mice in varies ages. Moreover, chitnase1 increased significantly in APP/PS1 mice aged 12 and 22 months compared with the age matched wild-type group, respectively. However, no difference of chitnase1 was found between 4 months-old APP/PS1 mice and wild-type mice. Comparing with the age matched wild type group, the consequences of mRNA on the increase in chitnase1 is in accordance with protein in APP/PS1 mice. Furthermore, Morris water maze showed that 4 months-old APP/PS1 mice have normal spatial learning and impaired spatial memory; both spatial learning and spatial memory in 12 and 22 months-old APP/PS1 mice were declined. Time-dependent increase of chitnase1 in APP/PS1 double transgenic mice indicates that the level of chitinase1 is associated with decline of cognition. Therefore, chitinase1 might be a biomarker of disease progression in AD.     

Mithramycin A Alleviates Cognitive Deficits and Reduces Neuropathology in a Transgenic Mouse Model of Alzheimer’s Disease

Increasing evidence has shown that specificity protein 1 (Sp1) is abnormally increased in the brains of subjects with Alzheimer’s disease (AD) and transgenic AD models. However, whether the Sp1 activation plays a critical role in the AD pathogenesis and selective inhibition of Sp1 activation may have a disease-modifying effect on the AD-like phenotypes remain elusive. In this study, we reported that Sp1 mRNA and protein expression were markedly increased in the brain of APPswe/PS1dE9 transgenic mice, whereas chronic administration of mithramycin A (MTM), a selective Sp1 inhibitor, potently inhibited Sp1 activation in the APPswe/PS1dE9 mice down to the levels of wild-type mice. Specifically, we found that MTM treatment resulted in a significant improvement of learning and memory deficits, a dramatic reduction in cerebral Aβ levels and plaque burden, a profound reduction in tau hyperphosphorylation, and a marked increase in synaptic marker in the APPswe/PS1dE9 mice. In addition, MTM treatment was powerfully effective in inhibiting amyloid precursor protein (APP) processing via suppressing APP, beta-site APP cleaving enzyme 1 (BACE1), and presenilin-1 (PS1) mRNA and protein expression to preclude Aβ production in the APPswe/PS1dE9 mice. Furthermore, MTM treatment strongly inhibited phosphorylated CDK5 and GSK3β signal pathways to reduce tau hyperphosphorylation in the APPswe/PS1dE9 mice. Collectively, our findings provide evidence that Sp1 activation may contribute to the AD pathogenesis and may serve as a novel therapeutic target in the treatment of AD. The present study highlights that selective Sp1 inhibitors may be considered as disease-modifying therapeutic agents for AD.     

Imbalance of Microglial TLR4/TREM2 in LPS-Treated APP/PS1 Transgenic Mice: A Potential Link Between Alzheimer’s Disease and Systemic Inflammation

Clinically, superimposed systemic inflammation generally has significant deleterious effects on the Alzheimer’s disease (AD) progression. However, the related molecular mechanisms remain poorly understood. Microglial toll-like receptor 4 (TLR4) and triggering receptor expressed on myeloid cells 2 (TREM2) are two key regulators of inflammation that may play an essential role in this complex pathophysiological process. In this study, intraperitoneal injection of lipopolysaccharide (LPS) into APP/PS1 transgenic AD model was used to mimic systemic inflammation in the development of AD. Initial results from the cortex showed that compared with wild-type mice, APP/PS1 mice exhibited elevated gene and protein expression levels of both TLR4 and TREM2 with different degree. Interestingly, after LPS treatment, TLR4 expression was persistently up-regulated, while TREM2 expression was significantly down-regulated in APP/PS1 mice, suggesting that the negative regulatory effect of TREM2 on inflammation might be suppressed by LPS-induced hyperactive TLR4. This imbalance of TLR4/TREM2 contributed to microglial over-activation, followed by increased neuronal apoptosis in the cortex of APP/PS1 mice; these changes did not alter the expression level of Aβ_1?42. Similar alterations were observed in our in vitro experiment with β-amyloid_1–42 (Aβ_1–42)-treated N9 microglia. Further, Morris water maze (MWM) testing data indicated that LPS administration acutely aggravated cognitive impairment in APP/PS1 mice, suggesting that the addition of systemic inflammation can potentially accelerate the progression of AD. Collectively, we conclude that an imbalance of TLR4/TREM2 may be a potential link between AD and systemic inflammation. TREM2 can serve as a potential therapeutic target for treating systemic inflammation in AD progression.

     

Presenilin-interacting proteins

Familial Alzheimer's disease (FAD) accounts for 5-10% of deaths from Alzheimer's disease (AD), and approximately 50% of these cases have been definitely linked to missense mutations in three genes, encoding the amyloid precursor protein (APP), presenilin 1 (PS1) and presenilin 2 (PS2). Of these, the vast majority of FAD-linked mutations are within PS1. There has been an extensive effort to identify proteins that functionally interact with PS1 and PS2 because of their clear roles in FAD. The goal of this review is to describe these proteins and to discuss in more detail the probable biological functions of a subset of the better-studied interacting proteins. In particular, the review examines APP, Notch, nicastrin, modifier of cellular adhesion (MOCA), beta-catenin, and the group of proteins involved in cell death, calcium metabolism and cell adhesion. We argue that, although a few of the interacting proteins are unambiguously involved in well-studied cellular pathways, their exact roles within these pathways have not been clearly defined, and indeed might vary between cell types. We also question the physiological relevance of some of the work linking PS to cell death pathways. Finally, we point out the value of using flies and worms to sort out the often contradictory work in the PS field, and we mention how knowledge of PS-interacting pathways will contribute to the development of new therapeutic strategies in AD.     

IGF2 Ameliorates Amyloidosis,Increases Cholinergic Marker Expression and Raises BMP9 and Neurotrophin Levels in the Hippocampus of the APPswePS1dE9 Alzheimer’s Disease Model Mice

The development of an effective therapy for Alzheimer’s disease (AD) is a major challenge to biomedical sciences. Because much of early AD pathophysiology includes hippocampal abnormalities, a viable treatment strategy might be to use trophic factors that support hippocampal integrity and function. IGF2 is an attractive candidate as it acts in the hippocampus to enhance memory consolidation, stimulate adult neurogenesis and upregulate cholinergic marker expression and acetylcholine (ACh) release. We performed a seven-day intracerebroventricular infusion of IGF2 in transgenic APPswe.PS1dE9 AD model mice that express green fluorescent protein in cholinergic neurons (APP.PS1/CHGFP) and in wild type WT/CHGFP littermates at 6 months of age representing early AD-like disease. IGF2 reduced the number of hippocampal Aβ40- and Aβ42-positive amyloid plaques in APP.PS1/CHGFP mice. Moreover, IGF2 increased hippocampal protein levels of the ACh-synthesizing enzyme, choline acetyltransferase in both WT/CHGFP and APP.PS1/CHGFP mice. The latter effect was likely mediated by increased protein expression of the cholinergic differentiating factor, BMP9, observed in IGF2-treated mice as compared to controls. IGF2 also increased the protein levels of hippocampal NGF, BDNF, NT3 and IGF1 and of doublecortin, a marker of neurogenesis. These data show that IGF2 administration is effective in reversing and preventing several pathophysiologic processes associated with AD and suggest that IGF2 may constitute a therapeutic target for AD.     

Processing of the Platelet Amyloid Precursor Protein in the Mild Cognitive Impairment (MCI)

It has been suggested that mild cognitive impairment (MCI) patients deteriorate faster than the healthy elderly population and have an increased risk of developing dementia. Certain blood molecular biomarkers have been identified as prognostic markers in Alzheimer’s disease (AD). The present study was aimed to assess the status of the platelet amyloid precursor protein (APP) metabolism in MCI and AD subjects and establish to what extent any variation could have a prognostic value suggestive of predictive AD in MCI patients. Thirty-four subjects diagnosed with MCI and 45 subjects with AD were compared to 28 healthy elderly individuals for assessing for protein levels of APP, β-APP cleaving enzyme 1 (BACE1), presenilin 1 (PS1) and a disintegrin and metalloproteinase-10 (ADAM-10) by western blot, and for the enzyme activities of BACE1 and γ-secretase by using specific fluorogenic substrates, in samples of platelets. A similar pattern in the healthy elderly and MCI patients was found for BACE1 and PS1 levels. A reduction of APP levels in MCI and AD patients compared with healthy elderly individuals was found. Augmented levels of ADAM-10 in both MCI and AD were displayed in comparison with age-matched control subjects. The ratio ADAM-10/BACE1 was higher for the MCI group versus AD group. Whereas BACE1 and PS1 levels were only increased in AD regarding to controls, BACE1 and γ-secretase activities augmented significantly in both MCI and AD groups. Finally, differences and similarities between MCI and AD patients were observed in several markers of platelet APP processing. Larger sample sets from diverse populations need to be analyzed to define a signature for the presence of MCI or AD pathology and to early detect AD at the MCI stage.     

PS-2基因的克隆及其在肝癌中的表达

利用荧光差异显示技术比较了正常肝、肝硬化和肝癌组织 m RNA的表达 ,1 4个有差异的条带通过 Northern blot分析表明其中 9个为阳性 .令人感兴趣的是一个～ 5 0 0 bp的 c DNA片段 ,它在正常肝和肝硬化中低表达 ,在肝癌组织中高表达 .通过测序 ,发现该片段与 PS- 2 ( presenilin- 2 )基因有 94 %的同源性 .PS- 2基因的突变与早发性阿尔茨海默氏症有关 ,但在肝癌发生中的作用未明 .也许 PS- 2基因的上调涉及到肝癌发生的分子机理