临床样本DNA自动测序结果的判读

自动测序技术向临床医学方向转化,使测序结果电泳图的正确判读成为临床分子病理学家、肿瘤学家及临床研究者十分重要的技能之一。本文总结了自动测序结果图中单位点杂峰、杂合子、纯合子及长片段杂峰序列,以及缺失与插入所致框移等的特点和正确判读方法,并就其发生原理、注意事项及处理方法等作一探讨。另外,结合临床实践对ＤＮＡ测序的报告内容提出了相应的要求。     

胃癌石蜡包埋组织的miRNA实时荧光定量PCR表达

目的：研究miRNA在胃癌石蜡包埋组织和新鲜组织的表达。方法：应用实时荧光定量PCR检测6例石蜡包埋胃癌组织及其正常胃黏膜组织、新鲜胃癌组织及其正常黏膜组织的miRNA表达（let-7a,miR-21,miR-155）,以U-6为内参对照。结果：胃癌石蜡包埋组织与胃癌新鲜组织中miRNA表达呈平行性高表达。结论：可以利用石蜡组织进行miRNA相关指标表达研究。     

Experimental therapeutic effect of combined cytokines on beagle dogs exposed to 4.5 Gy γ-rays

Objective To evaluate the effects of combined administration of recombinant human interleukin-11(rhIL-11),recombinant human G-CSF(rhG-CSF)and recombinant human interleukin-2 (rhIL-2)on acute radiation sickness(ARS)beagles.Methods Sixteen beagle were irradiated with 4.5 Gy60 Co γ-rays to establish ARS models,and were divided into irradiation control group,supportive care group and combined cytokines treatment group.After irradiation irradiation control group was given no treatment,the dogs in supportive care group received purely symptomatic treatment,while combined cytokines treatment group received rhIL-11 50μg/(kg·d)and rbG-CSF 10μg/(kg·d)subcutaneously(0-14 d)and rhIL-2 1×1 06 U/d(29-43 d)besides symptomatic treatment.Manifestation and characteristics of ARS beagles were observed,and the survival time were recorded.At last,post-mortem examination and histological examination were performed.Results All animals underwent nausea,diarrhea and fever.After irradiation,all animals in irradiation control group died in two weeks,and the mean survival time was 12.7 d,while only one died at 33 d in supportive care group.All dogs in combined cytokine group survived at 45 day after exposure,and their haematopoiesis and gastrointestinal tract were recovered.Conclusions Combination of rhIL-11 + rhG-CSF + rhIL-2 treatment could be significantly effective on ARS beagles irradiated by 4.5 Gy60 Co γ-rays,which could accelerate injured haemotopoiesis and intestinal tract recovery,increase the survival rate and improve the life quality of animals.     

Clotting mechanism in beagles irradiated by 4.5 Gy γ-rays

Objective To explore the clotting mechanism in beagles irradiated by 4.5 Gy γ-rays after treatment with supportive care,or supportive care and combined cytokines.Methods Sixteen beagles were divided into irradiation control group,Supportive care group and combined cytokines treatment group.Platelet aggregation test,thrombelagtography (TEG) and the time measurement were analyzed in vitro.Results In irradiation group and supportive care group,the platelet aggregation rates in beagles were decreased markedly and the k value of TEG was increased 7 d post-irradiation,while those indexes in combined cytokines treatment group changed little.At 14 d post-irradiation,each parameter of TEG in irradiated group changed obviously.The values of r,k,r+k and M were elevated significantly,clotting time and the maximum coagulation time of thrombus delayed,the Ma value was decreased markedly,and the maximum elasticity amplitude of thrombus was diminished.All parameters in combined cytokines treatment group were better than those in supportive care group.The thrombin time was prolonged obviously in irradiated group 14 d post-irradiation,while the thrombin time was the longest at 2-3 weeks post irradiation in supportive care group and combined cytokines treatment group(P>0.05).Conclusions Cytokines could improve the platelet aggregation and the blood clotting functions of beagles suffering from acute radiation sickness.     

γ线照射小鼠胸腺组织基因表达转录谱分析

目的 探讨6 Gy γ线照射后不同时间小鼠胸腺组织基因表达转录谱的动态变化。方法 应用快速高通量的cDNA基因芯片技术进行研究。结果 1随照射后时间推移,差异表达基因数目和种类逐渐减少,如照射后1、6、14和28 d差异基因数量分别为878、318、162和25;2辐射所诱导的胸腺组织差异表达基因广泛涉及细胞周期、免疫和应激、细胞凋亡、细胞信号转导、转录调节、DNA合成修复和重组、细胞骨架、离子通道和运输、代谢、蛋白翻译和合成、发育、细胞分化多个方面;3照射后对某些重要差异表达基因的分析表明,与细胞周期相关的基因有5个(上调3个:Cyclin G、Anxa1、Fgf1,下调2个:Cdc2a、Cdc25b);与免疫应激相关的5个(上调4个:IL-18、Casp1、IL-15、IL-7,下调1个:Cd28);与细胞凋亡相关的7个(上调4个:Casp1、Anxa1、Perp、IL-7,下调3个:Pten、Api5、Fas)。结论 6 Gy γ线照射后,小鼠胸腺组织差异表达基因随照射后时间延长不仅涉及多个靶点、多个层次,显示出多样性变化的特点,而且显示出时间上的明显差异;从基因水平上揭示了中等剂量照射小鼠胸腺组织损伤修复和重建基因的变化规律,为辐射免疫损伤与修复的分子机制和防治措施的研究提供了新的启示。     

α粒子诱发 BEP2D细胞转化过程中肺癌相关基因表达的 cDNA Microarray研究

目的：探讨辐射诱发人支气管上皮细胞（BEP2D）转化过程中肺癌相关基因的表达。方法：用Cartesian PixSys5500cDNA Microarray点样仪将60个肺癌相关基因以微阵列形式点布于醛基化的玻璃片上。提取α离子辐射前BEP2D细胞（原代）和α粒子辐射后20代、35代细胞总RNA,经长片段反转录和线性扩增标记成荧光探针后与微阵列中cDNA进行杂交。结果：原代细胞中检测到40个基因表达;20代检测到47个基因表达;35代检测到20个基因表达。所检测的基因中,抑癌基因的mRNA丰度的原代和20代后细胞中急剧下降;大多数癌基因的表达丰度在20代以后细胞中仅轻微下降;生长因子类基因大都在20代细胞表达。结论：在辐射诱发的人支气管上皮转化细胞中,抑癌基因的失活可能与细胞恶化有关,癌基因及生长因子类基因可能促进了细胞的转化。     

复方甲硝唑漱口液配方的改进

复方甲硝唑漱口液是一种医院自制复方制剂,由甲硝唑、醋酸氯己定、薄荷脑、糖精钠、尼泊金组成,用于防止口腔感染,含量测定采用高效液相法,但由于医院目前没有高效液相色谱仪,因此含量测定实际不能进行.我们试验改用双波长分光光度法进行含量测定,发现尼泊金和糖精钠在220～400nm波长处有干扰,针对这种情况,我们对配方进行了改进,去掉配方中的尼泊金,并将糖精钠换成甜菊素,避免了含量测定的干扰.为保证改进处方后的漱口液也具有良好的杀菌消炎作用,我们将按原配方和按改进后的配方制得的制剂进行卫生学对比实验,进行考察,结果如下.     

白血病样非淋巴结性套细胞淋巴瘤临床病理观察

目的 观察白血病样非淋巴结性套细胞淋巴瘤(LNNMCL)临床病理特征。方法 收集2例LNNMCL患者临床及实验室检查资料,取骨髓或脾脏组织进行病理检查,利用HE染色、免疫组化等方法明确诊断后治疗并随访,结合文献分析疾病的临床病理特点。结果 2例均为老年患者,实验室检查外周血淋巴细胞绝对计数显著增高,影像学检查脾大,未检测到肿大淋巴结,血涂片内检测到小到中等大淋巴细胞,核圆或椭圆,深染,可见凹陷及小核仁,流式检测提示肿瘤细胞表达B系表型;病理检查肿瘤淋巴细胞免疫组化阳性表达CD20、CD79a、PAX-5、CD5、CD43、CyclinD1,阴性表达SOX11、Bcl-6、CD10,细胞增殖活性较低,Ki-67增殖指数约5%;一例患者外周血肿瘤细胞核型分析染色体为非复合核型伴t(11;14)(q13;q32)染色体易位,另一例患者在脾脏组织中用二代测序检测出IgH/CylinD1基因易位,2例基因检测均显示IgHV及P53突变。综合临床、形态、免疫表型及基因检测结果,诊断为白血病样非淋巴结性套细胞淋巴瘤。采用化疗和靶向治疗后,分别随访69、12个月,患者病情稳定,健康状况良好。结论 白血...     

黄连温胆汤加减治疗中风52例

笔者用黄连温胆汤加减着重从痰论治本病52例,取得满意疗效,现报道如下: 1 临床资料 中风患者104例,年龄50～70岁,男81例,女23例,其中脑梗死82例,脑出血10例,脑栓塞12例,分为治疗组和对照组各52例.诊断参照全国第3次脑血管学术会议所定标准.     

杜冷丁配伍东莨菪碱加速产程进展的临床观察

目的探讨初产妇临产后杜冷丁配伍东莨菪碱一次性肌肉注射,对产程进展情况及新生儿的影响.方法 80例初产妇随机分为实验组、对照组各40例,实验组临产后(宫口开大2～3cm)时肌肉注射杜冷丁+东莨菪碱1次,对照组临产后肌肉注射杜冷丁1次.结果实验组用药至宫口开全时间为(360±33.1)min,对照组为(470±65.2)min(P＜0.05),实验组总产程时间(412.9±55.7)min,对照组为(511.9±102.8)min,(P＜0.05).两组新生儿Apgar评分经统计学分析有显著性意义.结论临产后使用杜冷丁+东莨菪碱肌肉注射能缩短产程、对新生儿无不良影响.