肺癌癌组织中M2-PK、和ENO1的表达水平及对其生存期的预测价值

目的研究肺癌组织中肿瘤型丙酮酸激酶(M2-PK)和烯醇化酶-α(ENO1)的表达对患者生存期的预测价值。方法采用免疫组织化学染色法分别对182例肺癌患者和68例肺良性病变组织切片进行检测,观察M2-PK和ENO1在肺癌组织和良性病变组织中的表达情况,分析M2-PK和ENO1表达情况与肺癌临床病理特征及患者生存期的关系。结果肺癌组织中M2-PK和ENO1阳性表达率分别为78.02%、67.58%,高于良性病变组织的32.36%和29.41%,差异具有统计学意义(P<0.001),其表达与肺癌浸润分期(P=0.027,P=0.019)、分化程度(P=0.041,P=0.037)、淋巴转移(P=0.009,P=0.001)、TNM分期均显著相关(P=0.012,P=0.015);肺癌组织中M2-PK与ENO1表达呈正相关(r=0.769,P<0.05);肺癌组织中M2-PK和ENO1的高表达与患者5存活率分别为37.35%和32.31%,低于低表达的62.62%和52.99%,差异具有统计学意义(P<0.05);M2-PK与ENO1共高表达患者5年存活率为28.84%,低于共低表达的59.32%(P<0.05)。结论肺癌组织中M2-PK和ENO1的高表达与肺癌的发生、发展及预测患者的生存期有一定的相关性,可能存在临床应用价值。     

Glu29→Gly纯合子突变导致的遗传性凝血酶原缺乏症

目的 对1个遗传性凝血酶原缺乏症家系进行凝血酶原(FⅡ)基因突变的检测。方法 用活化的部分凝血活酶时间（APTT)，凝血酶原时间(PT)及FⅡ促凝活性(FⅡ：C)、FⅡ抗原(FⅡ：Ag)测定进行表型诊断；用PCR法对先证的FⅡ基因14个外显子及其侧翼序列和5′端非翻译区(5′UTR)、3′端非翻译区(3′UTR)序列进行扩增，PCR产物纯化后直接测序，检测其基因突变。家系成员DNA在先证FⅡ基因突变区域扩增后测序。突变位点经限制性内切酶分析证实。103名健康献血作对照。结果 先证表型诊断为凝血酶原缺乏症(Ⅰ型)；FⅡ外显子区共发现3个与献报道的FⅡ基因序列不同的位点，其中位于第2外显子区的为纯合突变A601G。家系分析表明先证父亲、母亲和外祖母均为A601G杂合子。结论 纯合错义突变A601G引起的Glu29→Gly是导致该例遗传性凝血酶原缺乏的原因。     

广州地区医院获得性肺炎耐药性监测

目的明确广州地区医院获得性肺炎(HAP)的病原菌构成及耐药情况。方法2004年1月-2005年10月,在广州市6所三级甲等医院发生HAP的病原菌进行鉴定,纸片扩散法检测其耐药性。结果HAP前8种病原菌分别为铜绿假单胞菌、金黄色葡萄球菌、肺炎克雷伯菌、白色假丝酵母菌、嗜麦芽寡养单胞菌、鲍氏不动杆菌、洋葱伯克霍尔德菌、大肠埃希菌等;革兰阳性球菌和革兰阴性杆菌均严重耐药,耐甲氧西林金黄色葡萄球菌占91.8%,溶血葡萄球菌全部耐甲氧西林。结论广州地区HAP耐药严重,必须加强防治措施。     

两个特殊血友病A患者家系的基因诊断

目的 探讨特殊血友病A(HA)家系的基因诊断。方法 用内含子22倒位检测及FⅧ基因内外4个位点的多态性进行遗传连锁分析;用核酸直接测序法对FⅧ各外显子及其侧翼序列进行检测。结果 家系1先证者表型为重型HA,FⅧ内含子22倒位检测及FⅧ基因内外4个位点的多态性遗传连锁分析未获得诊断信息;直接核酸测序证实先证者及其妹FⅧ基因24号外显子第2209位氨基酸存在R2209Q错义突变,后者胎儿DNA检测为正常男性。家系2中HA先证者已去世,一名要求做携带者检测的女性表型正常,内含子22倒位检测及直接核酸测序没有发现基因缺陷,用FⅧ基因内外4个位点的多态性遗传连锁分析均显示该成员未携带血友病A基因。结论 联合应用直接核酸测序法及多态性遗传连锁分析可以对特殊血友病A家系进行基因诊断。     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

重症社区获得性肺炎炎性机制研究进展

重症社区获得性肺炎是人类重要的死亡原因之一，也是目前临床所关注的问题。重症社区获得性肺炎一般是严重的细菌或病毒感染所致，因此临床对抗感染和抗病毒治疗十分重视。但随着对该疾病的进一步认识，重症社区获得性肺炎产生的全身炎症反应及进而导致的多器发功能障碍综合征(MODS)是机体防御机制的过度激活而引起自身破坏的结果，而不是细菌、毒素等直接损伤的结果，     

593例医院感染现患率调查分析

目的 了解医院感染现状以及抗菌药物使用情况,为制定防控措施提供科学依据。方法 采取床旁调查和住院病历调查相结合,填写床旁调查表及个案登记表,对2010年6月3日的住院患者进行医院感染现患率调查。 结果 实查593例,实查率100%。医院感染例次率为5.40%(32/593)。感染部位按部位感染率由高至低依次为下呼吸道、创面、上呼吸道、表浅切口。病原体学送检率为43.75%,抗菌药物使用率42.66%。 结论 加强医院感染管理,规范抗菌药物的合理使用,从而降低医院感染率。     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

腺病毒E1A蛋白抑制大鼠肺泡上皮细胞γ-谷氨酰半胱氨酸合成酶催化亚单位表达的机制研究

目的： 研究腺病毒E1A蛋白抑制大鼠肺泡上皮细胞γ-谷氨酰半胱氨酸合成酶催化亚单位（GCLC）表达的机制。方法: 构建稳定表达腺病毒E1A蛋白的大鼠肺泡上皮细胞，将GCLC基因5’-上游调控序列不同长度的缺失体-萤火虫荧光素酶报道系统转染后分析转录活性的变化；检测AP-1、NF-κB、USF与GCLC基因调控序列的结合活性。结果: GCLC基因5’-上游调控序列报道系统检测结果显示大部分（9/11）缺失体的转录活性抑制，AP-1、NF-κB、USF与GCLC基因的结合活性增强，而对应的功能元件的转录活性降低。结论: 腺病毒E1A蛋白通过抑制GCLC基因 5’-上游调控序列的转录活性，扩大大鼠肺泡上皮细胞氧化应激时的氧化/抗氧化失衡,其机制可能涉及E1A对辅助转录因子的抑制。