K-ras基因突变与结直肠癌临床病理特征的相关性研究

目的:探讨结直肠癌患者K-ras基因突变特点,并分析K-ras基因突变与结肠癌临床病理特征的相关性及其临床意义。方法:收集哈尔滨医科大学附属第一医院收治的500例结直肠癌患者的手术石蜡标本,采用突变扩增阻滞系统技术(amplification refractory mutation system,ARMS)检测K-ras基因第二外显子第12、13密码子的七种热点突变情况,分析其与结肠癌患者临床病理特征的相关性。结果:ARMS法检出K-ras基因突变213例,突变率为42.6%。K-ras基因突变与患者的年龄、性别、肿瘤发生的部位、肿瘤分化程度及有无淋巴结转移均无相关性(P0.05),而与肿瘤的组织学类型具有相关性(P0.05)。结论:(1)结直肠癌患者K-ras基因突变率较高,其中GLY12ASP位点突变率最高,GLY12ARG为稀有突变位点。(2)K-ras基因突变与结肠癌的组织学类型相关,可能用于结直肠癌患者治疗方案的选择。     

非小细胞肺癌患者K-ras基因突变与预后的相关性分析

目的:探讨非小细胞肺癌(NSCLC)患者K-ras基因突变与预后的相关性。方法:回顾分析2006年3月~2008年7月江西省肿瘤医院收治的92例NSCLC患者的临床资料,所有患者均采用蝎形探针扩增阻遏突变系统(ARMS)法行K-ras基因突变检测。结果:92例NSCLC患者中,11例K-ras基因突变阳性,阳性率为11.96%,包括Gly12Cys(GGTTGT)突变3例、Gly12Val(GGTGTT)突变2例、Gly12Ala(GGTGCT)突变2例、Gly12Arg(GGTCGT)突变2例、Gly12Cys(GGTTGT)突变与Gly12Val(GGTGTT)突变并存1例、Gly12Cys(GGTTGT)突变与Gly12Ala(GGTGCT)突变并存1例;K-ras基因突变阳性组与阴性组之间的患者性别、年龄、病理类型、吸烟史相比,差异无统计学意义(P0.05);K-ras基因突变阳性组Ⅲa~Ⅳ期患者中位生存期显著低于K-ras基因突变阴性组Ⅲa~Ⅳ期患者,差异有统计学意义(P0.05)。结论:NSCLC患者K-ras基因突变阳性率较低,但它可以对NSCLC患者预后产生负性影响。     

超声内镜引导下细针穿刺活检联合基因检测对胰腺癌的诊断价值

目的:研究超声内镜引导下细针穿刺活检(EUS-FNA)联合K-ras基因检测对胰腺癌的诊断价值,为临床诊疗提供依据。方法:选取2013年11月到2015年11月我院收治的胰腺占位病变患者90例,患者入院次日行EUS-FNA,检测患者血清及活检物中K-ras基因阳性率,比较EUS-FNA单独与EUS-FNA联合K-ras基因检测对胰腺癌诊断的准确率与敏感性。结果:90例胰腺占位病变者中,经病理证实胰腺癌56例,EUS-FNA单独与EUS-FNA联合K-ras基因分别检出胰腺癌50例、53例,准确率分别为89.29%、94.64%,敏感性分别为92.59%、98.15%,两组比较差异均有统计学意义(P0.05)。胰腺癌患者活检物中K-ras阳性率为83.93%,明显高于血清中的41.07%(P0.05)。结论:EUS-FNA联合K-ras基因检测可提高对胰腺癌诊断的准确率与敏感性。     

肺腺癌组织中表皮生长因子受体基因突变的检测及其临床意义

目的:通过检测肺腺癌组织中表皮生长因子受体(EGFR)基因的突变情况,研究EGFR突变与患者临床特征(性别、年龄和吸烟史)的相关性。方法:收集160例肺腺癌患者术前的石蜡组织标本,提取DNA后用实时定量PCR方法对EGFR基因18~21外显子进行突变检测;对基因突变结果与患者的性别、年龄和吸烟史分别做χ2检验。结果:160例肺腺癌组织标本中,有57例检测到EGFR基因突变,突变率为35.6%,且突变与患者性别、年龄和吸烟史均无显著相关性(P0.05);57例EGFR基因突变标本中,27例为19外显子缺失,22例为21外显子L858R点突变,这2种突变占总突变类型的85.96%,并且与患者性别、年龄及吸烟史无显著相关性(P0.05)。结论:EGFR基因在女性不吸烟肺腺癌中有较高的突变率,突变主要集中在19外显子缺失和21外显子L858R点突变,但突变率及突变类型与患者性别、年龄和吸烟史均不相关。     

胃癌中Fascin 表达与K-ras 突变情况及两者相关性的研究

目的：观察胃癌组织中Fascin 表达和K-ras 突变情况,并分析其与临床病理特征的关系,研究Fascin表达和K-ras突变之间 的相关性,探讨两者在胃癌浸润转移过程中的作用。方法：在108 例胃癌组织中采用免疫组织化学法检测Fascin 表达,采用DNA 测序法检测K-ras突变。结果：108 例胃癌组织中Fascin 表达率为25.00 %,K-ras突变率为11.11 %;胃癌组织中Fascin 阳性表达, 发生淋巴结转移组多于未发生淋巴结转移组(P＜0.05),发生远处转移组多于未发生远处转移组（P＜0.01）;K-ras 突变,发生远处 转移组多于未发生远处转移组（P＜0.01）;胃癌组织中K-ras 突变和Fascin 表达之间存在相关性,Fascin阳性比Fascin阴性的胃癌 组织更倾向于发生K-ras突变（rp=0.378,P＜0.01）。结论：在胃癌组织中K-ras 基因突变和Fascin蛋白阳性表达存在相关性,且与 肿瘤的转移有关。     

非综合征型耳聋患者耳聋易感基因突变的检测分析

目的:探究非综合征型耳聋患者耳聋易感基因的携带情况及突变类型,为耳聋患者治疗或遗传咨询提供理论依据。方法:收集821例非综合征型耳聋患者的外周静脉血,提取基因组DNA后,进行4个常见耳聋易感基因GJB2、GJB3、SLC26A4和线粒体12S r RNA的9个突变热点筛查。结果:821例非综合征型耳聋患者中耳聋易感基因筛查阳性375例,阳性率为45.7%,不同性别之间阳性率无明差异(P=0.625)。375例存在耳聋易感基因突变的研究对象中,4个易感基因的9突变热点共检测出447例点突变,其中GJB2基因的有241例点突变(53.9%),以235 del C位点突变率最高;SLC26A4基因的有126例点突变(28.2%),以IVS7-2 AG位点突变为主;线粒体12S r RNA基因的有79例点突变(17.7%),绝大部分为1555 AG位点突变;而GJB3基因仅有1例点突变(0.2%)。375例存在耳聋易感基因突变的研究对象中,304例发生1种突变(81.1%),有70例发生2种突变(18.7%),仅有1例发生3种突变(0.3%)。结论:非综合征型耳聋患者中GJB2基因的235 del C位点以及SLC26A4基因的IVS7-2 AG位点突变率较高,常见耳聋易感基因筛查有助于非综合征型耳聋患者的诊断、干预及治疗。     

HPV16 变异体在新疆地区宫颈癌及癌前病变中的分布研究

目的:研究新疆地区HPV16 E6、E7、LCR基因突变情况,分析HPV16变异体在宫颈癌及癌前病变发生发展中的作用。方法:选择HPV16阳性的宫颈癌及癌前病变患者,提取基因组DNA,利用PCR扩增HPV16 DNA E6、E7基因及LCR区核苷酸片段,正反向测序。与HPV16基因序列分析比对,分析核苷酸突变位点。结果:E6基因突变率为80.00%(92/115)主要突变位点T350G(59.78%)、T178G(18.47%);E7突变率为54.78%(63/115),主要突变位点A647G(33.33%)、T846C(26.98%);LCR突变率为23.48%(27/115),主要突变位点为C24T(74.07%)、C13T(25.92%)。维吾尔族T350G突变率较汉族妇女显著升高,而汉族A647G、T846C、C24T突变率显著高于维吾尔族,差异具有统计学意义(P0.05)。维吾尔族宫颈癌组T350G突变率显著高于炎症组(P0.05),且随病变严重程度增加突变率上升,汉族T350G、A647G、T846C、C24T突变率炎症组、宫颈病变组显著高于宫颈癌组(P0.05),维吾尔族C24T突变率炎症组显著高于宫颈癌组(P0.05),差异均具有统计学意义(P0.05)。结论:HPV16E6、E7突变可能与宫颈病变进展有关,T350G突变可能是维吾尔族宫颈癌高发的原因之一。     

膀胱癌H-ras基因点突变和ras癌基因产物P21的表达

应用聚合酶链反应(PCR)技术结合地高辛化寡核甘酸探针杂交以及免疫组织化学方法,检测人膀眈癌组织中H-ras癌基因第12位密码子点突变和基因产物P21蛋白的表达.58例膀胱癌组织中有23例出现H-ras基因的点突变,36例P21蛋白表达阳性,5例正常膀胱组织未见点突变和P21蛋白阳性表达.本组膀胱癌第12位密码子点突变和P21阳性表达水平呈正相关,多发生在肿瘤临床T2-T4期和分化差的肿瘤,并且与膀胱癌的预后有关、提示H-ras癌基因点突变和P21蛋白阳性表达是膀胱癌的晚期表现,并在膀胱癌的发生,发展及预后中起着重要作用.     

Ⅰ-ⅢA期非小细胞肺癌KRAS基因突变与ERCC1、TYMS mRNA 表达水平的相关研究

目的:探讨Ⅰ-ⅢA期非小细胞肺癌(NSCLC)KRAS基因突变与核苷酸切除修复交叉互补基因1(ERCC1)、胸苷酸合成酶(TYMS)mRNA表达水平的相关性及其与患者临床病理特征的关系。方法:收集空军总医院胸外科2010年06月至2014年10月符合入组条件的Ⅰ-ⅢA期NSCLC患者69例,肺癌组织标本均为手术中切取,KRAS基因突变应用x TAG-液相芯片法检测,ERCC1、TYMS mRNA表达水平应用分支DNA-液相芯片法检测。结果:在69例检测样本中,共有13例存在KRAS基因突变,突变率为18.8%(13/69);男性患者中KRAS基因突变率(29.3%,12/41)较女性患者(3.6%,1/28)高(P=0.007)。ERCC1 mRNA的表达水平与病理类型、吸烟史、有无淋巴结转移、临床TNM分期相关(P0.05),TYMS mRNA表达水平与患者各临床病理特征无关(P0.05)。KRAS突变型患者ERCC1 mRNA表达水平比KRAS野生型患者高(P0.05),KRAS基因突变与TYMS mRNA表达水平无关(P0.05)。结论:在Ⅰ-ⅢA期NSCLC患者中,男性患者更容易发生KRAS突变。KRAS突变型患者可能不能从铂类化疗药物中受益,有利于指导早中期NSCLC患者术后的个体化治疗。     

110例维吾尔族及汉族乳腺癌中BRCA1基因突变及P53蛋白表达的研究

目的研究维吾尔族及汉族乳腺癌患者BRCA1基因突变及P53蛋白的表达。方法选取70例维吾尔族和40例汉族乳腺癌根治标本,对照组为32例维汉族乳腺良性病变(纤维腺病及纤维腺瘤)及乳腺癌旁非癌组织;运用PCR-SSCP和DNA序列测定的方法检测BRCA1基因突变及Evision二步法检测P53蛋白的表达。结果(1)110例维吾尔族及汉族乳腺癌中发现BRCA1突变的14个新位点。(2)110例维吾尔族及汉族乳腺癌BRCA1的突变率为10%,其中22例维吾尔族早发性乳腺癌(≤35岁)BRCA1突变率为31.82%,高于维吾尔族晚发性乳腺癌(P<0.01)。(3)110例维吾尔族及汉族乳腺癌中发现11例BRCA1基因核苷酸多态性位点。(4)BRCA1基因突变相关性乳腺癌中P53蛋白阳性表达率高于对照组,其淋巴结转移率高于对照组,其发病年龄小于对照组(P<0.01)。结论BRCA1基因突变与新疆维吾尔族早发性乳腺癌密切相关,且BRCA1突变相关性乳腺癌具有P53阳性率高、发病年龄趋于年轻化、淋巴结转移率高的趋势,这些特点有可能为基因检测前的筛选提供参考依据。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

医疗机构合理用药评价模型的构建研究

目的 针对医疗机构的合理用药水平进行评价研究。方法 根据医疗机构合理用药的具体要求,构建医疗机构合理用药评价指标体系,采用基于模糊群决策的方法和多指标评价分析法构建医疗机构合理用药评价模型。结果 构建了基于模糊群决策的医疗机构合理用药评价模型,并通过实例分析证明了评价模型的可行性。结论 建立的基于模糊群决策的医疗机构合理用药评价模型能够对医疗机构的合理用药水平进行科学评价,为提高医疗机构合理用药水平奠定基础。     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.