老年急性淋巴细胞白血病分子生物学改变的初步观察

目的 研究老年急性淋巴细胞白血病(ALL)的分子生物学特征。方法 应用多聚酶链反应技术(PCR)检测了16例老年ALL患者的bcr／abl融合基因、TCRγ基因重排(TCRγRA)以及IgH基因重排(IgHRA)。结果 16例患者中IgHRA11例，TCRγRA4例，两类基因重排均阴性1例；bcr／abl融合基因阳性9例，阳性率56．25％。结论 老年ALL以B—ALL为多，bcr／abl融合基因阳性率较高，可能是其治疗效果较差的原因之一。     

应用半重叠TCRγ引物扩增TCRγ基因重排检测淋巴细胞白血病的研究

采用更加敏感的半重叠式T细胞受体（TCR）γ链特异性引物的多聚酶链反应（PCR）法,对28例急性淋巴细胞白血病（ALL）初治、完全缓解（CR）及骨髓移植（BMT）患儿的骨髓标本进行检测,用消化煮沸及酚抽提法分别对所有骨髓标本进行DNA提取,将PCR结果进行比较。结果表明,消化煮沸法用于微量标本的DNA提取优于酚抽提法。28例ALL患儿中16例检出TCRγ特异性条带,其中微小残留病（MRD）组18例;阳性检出12例,其中免疫分型标记为B细胞型的4例（25.0%）,免疫标记为T细胞型者全部出现TCRγ阳性条带。表明TCRγ基因重排并非克隆性T细胞增生所特有,部分ALL患儿存在双克隆重排。     

426例急性白血病细胞免疫表型与临床特点

采用21种单克隆抗体以APAAP法测定426例急性白血病细胞的表面标记,还对31例急性淋巴细胞白血病（ALL）进行免疫球蛋白重链（IgH）、T细胞受体（TCRγ,δ）基因重排研究。结果:291例（68.5％）表达纯系表型,其中 T-ALL97例,B-ALL93例,TB-ALL35例,急性非淋巴细胞白血病（ANLL）66例;淋、髓多系表型14例（3.3％）;非系限表型93例（21.8％）;裸型28例（6.5％）。临床观察表明非系限型、裸型、B-ALLⅣ型缓解率低,预后差。免疫分型不仅提高白血病的确诊率,而且能指导临床治疗与判断预后。     

急性非淋巴细胞性白血病免疫球蛋白和T细胞受体基因重排的研究及其意义

采用PCR法对25例不同时期急性非淋巴细胞性白血病（ANLL）患者免疫球蛋白重链（IgH）和T细胞受体γ链（TcRr）基因重排进行了研究。结果显示，3例（12．0％）发生IgH基因重排，3例（12．0％）出现TcRr基因重排，其中1例同时出现IgH和TcRr基因重排，对这种ANLL中序列失真现象的机制及其在白血病基因分型及检测微小残留疾病中的意义进行讨论。     

儿童急性白血病形态学免疫学和细胞遗传学的分型诊断

目的：准确地进行儿童急性白血病（AL）的诊断分型,提高初诊患儿的诊断符合率。方法：采用形态学、免疫学和细胞遗传学（MIC）相结合的诊断方法,分析了110例初诊为AL的患儿。结果：形态学与MIC分型诊断符合率为88．2％;急性淋巴细胞性白血病（ALL）免疫分型诊断符合率为92．2％;而急性髓细胞性白血病（AML）仅为62．9％。8／35例AML表达淋系抗原（1y^ -AML),12／59例ALL表达髓系抗原（My^ -AML);11／110例为杂合性白血病。染色体核型异常检出率为63．6％。t(8;21）易位见于（13／21例）M2;t(7;11）易位见于12例M2;t（15;17）易位见于（2／5例）M3;t（9;22）和t（4;11）易位见于（8／64例）ALL。结论：运用MIC诊断分型方法能提高儿童AL的诊断率,为AL个体化治疗和评估预后提供信息。     

肺黏膜相关淋巴组织型淋巴瘤的表型和IgH基因重排的检测价值

目的探讨肺黏膜相关淋巴组织型淋巴瘤（MALTLoma）的免疫表型和免疫球蛋白重链（IgH）基因重排检测的意义。方法对12例肺MALTLoma患者的临床病理资料进行回顾性分析和免疫组化研究，7例进行IgH和T细胞受体γ链（TCRγ）基因重排检测，并对治疗后结果随访。结果12例中，开胸肺活检7例，经电视胸腔镜肺活检2例，细针肺穿刺活检3例。病理组织检查结果：瘤细胞主要由中心细胞样淋巴细胞、小淋巴细胞样瘤细胞组成；12例有淋巴上皮病变，11例有反应性淋巴滤泡，10例有淋巴滤泡的克隆化，9例有血管受侵，4例有胸膜受侵；瘤细胞表达B细胞相关抗原。7例行IgH基因重排检测，FR2阳性6例，FR3A阳性5例。TCRγ1和TCRγ2基因重排检测均为阴性。12例患者单纯化疗3例，手术切除8例，其中术后化疗6例，对症治疗1例；随访11例，随访时间1—12年，复发1例，死亡2例。结论组织病理学结合免疫组化检查能对大多数具有典型病变的肺MALTLoma进行诊断；在肺MALTLoma与肺良性淋巴组织增生性疾病的鉴别诊断上，IgH基因重排检测有重要的辅助价值。     

急性淋巴细胞白血病IgH和Vδ2Dδ3基因重排检测在微小残留病监测中的意义

目的 :探讨急性淋巴细胞白血病 (ALL)IgH和Vδ2 Dδ3基因重排分布频率及定量检测在微小残留病(MRD)监测中的意义。方法 :对初诊ALL患者及ALL完全缓解期 (CR)后不同时期的骨髓标本及脑脊液标本进行IgH和Vδ2 Dδ3基因重排检测及定量计算MRD值。结果 :1 0 2例初诊ALL患者骨髓标本IgH和Vδ2 Dδ3基因重排阳性率分别为 49.0 %和 36 .3 %。诱导缓解期与完全缓解期脑脊液IgH基因重排分别有 7例和 3例 ,Vδ2 Dδ3基因重排分别为 5例和 2例。MRD值 >0 .1 %者 3例 ,复发率为 66 .7%。MRD为 0 .0 0 2 %～ 0 .1 %者 6例 ,复发率为 1 7.0 % ;MRD <0 .0 0 2 %者 9例 ,无复发。复发组MRD明显高于未复发组 (P <0 .0 5)。结论 :动态PCR检测ALL患者脑脊液IgH和Vδ2 Dδ3基因重排比细胞学检测更灵敏。IgH和Vδ2 Dδ3基因重排MRD值增高 ,复发危险率增高。MRD定期定性定量监测对指导化疗药物的选择、疗效观察及判断预后有指导意义。     

慢性髓细胞白血病急变期形态学及免疫学和细胞遗传学特征研究

目的：探讨慢性髓细胞白血病急变期（CML－BC）的形态学、免疫表型和细胞遗传学法及流式细胞仪进行细胞免疫分型,细胞遗传滂采用直接法或短期培养法制备染色体标本,采用R显带技术进行核型分析。结果：免疫分型结果显示：急变为急性髓细胞白血病（AML）23例占74．2％;急性淋巴细胞白血病（ALL）5例占16．1％,均为B系ALL,其中4例同时伴有髓系表达;急性未分化细胞白血病1例,B系和髓系急性混合细胞白血病（AMLL）2例。31例CML－BC中21例（67．7％）的急变患者CD34^ ,其中4／5（80．0％）ALL,15／23（65。2％）,2／2AMLL均为CD34^ 。AML急变患者中具有CD7和CD34共表达者为8／23（占34．8％）。细胞遗传学分析表明,14／27（51．9％）和急变期患者出现Ph染色体以外的附加核型异常,其中有＋8（3／14）,＋Ph(3/14),i(17q)(2/14),Y染色体丢失（1／14）及复杂易位5／14）。结论：CML－BC是一干细胞疾病,原始细胞分化阻滞在早期阶段,故预后差。MIC分型在CML－BC诊断,预后判断及指导治疗方面均有重要价值。     

15例急性混合细胞白血病的临床与实验研究

目的：研究急性混合细胞白血病(MAL)的临床特征、实验室指标、治疗及预后。方法：对15例MAL骨髓标本分别进行光镜细胞形态学及相关细胞化学染色观察，以确定其FAB类型，应用流式细胞仪做免疫分型检测一系列相关单抗，同时采用G显带技术进行核型分析；应用PCR基因扩增方法检测TCRδ／lgH CDRⅢ2基因重排；运用急性粒细胞白血病(AML)、急性淋巴细胞白血病(ALL)或兼顾二者的治疗方案。结果：MAL患者的临床表现与AML及ALL患者的差异无显著性意义，形态学上MAL表现为AML的多为M2a、M1、M4，表现为ALL的多为ALL-L2；免疫分型显示MAL患者中以三系共表达者多见。另外，CDll7、HLA-DR在MAL中呈高表达。提示MAL的白血病细胞可能起源于早期造血细胞；核型分析可见异常染色体出现，但无特异性；基因重排显示部分患者TCRδ／lgH CDRⅢ2为阳性；患者对治疗反应差，生存期相对较短。结论：MAL具有独特的临床、实验室检查指标及预后。     

免疫球蛋白重链基因重排对多发性骨髓瘤的研究及应用价值

为探讨免疫球蛋白重链（IgH）基因重排在浆细胞增殖性疾病中的应用价值，用聚合酶链反应（PCR）技术以10例正常人骨髓标本为对照，对57例多发性骨髓瘤（MM）、11例未定性单克隆丙种球蛋白病（MGUS）、10例反应性浆细胞增多症进行了IgH基因重排的研究。MM的外周血和骨髓的IgH基因重排检出率分别为56．9％及84.4％。外周血IgH基因重排的分析显示Ⅱ、Ⅲ期患者检出率高于Ⅰ期。经化疗后缓解的病例其骨髓标本，仍可检出重排带。MGUS及反应性浆细胞增多症均未能测出IgH基因重排。本结果表明：IgH基因重排对MM的诊断、鉴别诊断及指导治疗有重要意义。     

妊娠时间与肺结核复发的相关性研究及诊治对策

目的 分析肺结核史患者妊娠时间和肺结核复发间相关性.方法 选取我院收治的有肺结核史的妊娠妇女576例作为研究对象,对其妊娠前肺结核治疗、治愈后妊娠时间、妊娠后复发肺结核等进行分析,总结有肺结核史育龄女性的妊娠时间和肺结核复发之间的关系.结果 肺结核治愈后不同时间段妊娠者的结核复发率比较,差异具有显著性（P＜0.05）,停药后间隔时间越久妊娠,肺结核复发的几率越小.结论 加强孕期痰菌检查,及早发现复发肺结核,提高母婴安全.     

我国骨关节结核诊治的回顾与展望

骨关节结核是危害人们健康的严重感染性疾病,近95％由他处结核病继发而来.罹患骨关节结核疾病后几乎均将致残,严重影响人们的健康、工作和生活.建国以来在党和国家的关心和支持下,骨关节结核的诊治水平取得了长足进步.时至今日,由于多种原因,学科发展和被重视程度受到一定的制约,同整个医疗行业的发展不相适应.回顾过去,展望未来,我们需要重新审视骨关节结核的诊治方法,努力推进骨关节结核诊疗技术的科学发展.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fos and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44~(MAPK), p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44~(MAPK), p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44~(MAPK) and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between P42/44~(MAPK) and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Raf/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44~(MAPK), c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

Overweight and Obesity and Progression of ADPKD

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Replication and Inhibitors of Enteroviruses and Parechoviruses

The Enterovirus (EV) and Parechovirus genera of the picornavirus family include many important human pathogens, including poliovirus, rhinovirus, EV-A71, EV-D68, and human parechoviruses (HPeV). They cause a wide variety of diseases, ranging from a simple common cold to life-threatening diseases such as encephalitis and myocarditis. At the moment, no antiviral therapy is available against these viruses and it is not feasible to develop vaccines against all EVs and HPeVs due to the great number of serotypes. Therefore, a lot of effort is being invested in the development of antiviral drugs. Both viral proteins and host proteins essential for virus replication can be used as targets for virus inhibitors. As such, a good understanding of the complex process of virus replication is pivotal in the design of antiviral strategies goes hand in hand with a good understanding of the complex process of virus replication. In this review, we will give an overview of the current state of knowledge of EV and HPeV replication and how this can be inhibited by small-molecule inhibitors.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fas and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44MAPK, p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44MAPK, p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44MAPK and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between p42/44MAPK and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Rat/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44MAPK, c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

心电图、心电向量图与冠状动脉造影结果对比分析

目的:通过分析心电图(Electrocardiogram,ECG)和心电向量图(Vectorcardiogram,VCG)的改变与冠脉造影（CAG）结果进行对比,探讨ECG、VCG在冠状动脉病变中的诊断价值。方法: 选择2008年1月~2009年12月临床拟诊断为冠心病患者108例,行常规ECG、VCG检查,并于1周内进行CAG,对检查结果依据各自的诊断标准进行判定,以CAG为标准诊断法,利用四格表法,计算相关评价真实性的指标并进行比较。结果: ①VCG检测的灵敏度、特异度、准确度显著高于ECG(P<0.05,P<0．01)。②ECG、VCG阳性率与冠脉病变支数组间比较:在单支病变、双支病变中,VCG阳性率明显高于ECG(P<0.05),左主干或三支病变无统计学意义;组内比较:ECG组左主干或三支病变组较单支病变、双支病变阳性率高(P<0.05,P<0.01);VCG组左主干或三支病变组较单支病变阳性率高(P<0.05);与双支病变阳性率比较无统计学意义;③ECG、VCG阳性率与冠脉病变程度组间比较:冠脉病变狭窄50%～69%的VCG阳性率明显高于ECG (P<0.05),其他两组阳性率比较无统计学意义;组内比较:ECG组冠脉病变狭窄≥90%较50%～69%、70%～89%的阳性率高(P<0.05,P<0.01); VCG组狭窄≥90%较50%～69%阳性率高（P<0.01),其他无统计学意义。结论: VCG对冠心病检测价值显著高于ECG。     

Detection and characterization of the product of hydroethidine and intracellular superoxide by HPLC and limitations of fluorescence

Here we report the structural characterization of the product formed from the reaction between hydroethidine (HE) and superoxide (O(2)(.-)). By using mass spectral and NMR techniques, the chemical structure of this product was determined as 2-hydroxyethidium (2-OH-E(+)). By using an authentic standard, we developed an HPLC approach to detect and quantitate the reaction product of HE and O(2)(.-) formed in bovine aortic endothelial cells after treatment with menadione or antimycin A to induce intracellular reactive oxygen species. Concomitantly, we used a spin trap, 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO), to detect and identify the structure of reactive oxygen species formed. BMPO trapped the O(2)(.-) that formed extracellularly and was detected as the BMPO-OH adduct during use of the EPR technique. BMPO, being cell-permeable, inhibited the intracellular formation of 2-OH-E(+). However, the intracellular BMPO spin adduct was not detected. The definitive characterization of the reaction product of O(2)(.-) with HE described here forms the basis of an unambiguous assay for intracellular detection and quantitation of O(2)(.-). Analysis of the fluorescence characteristics of ethidium (E(+)) and 2-OH-E(+) strongly suggests that the currently available fluorescence methodology is not suitable for quantitating intracellular O(2)(.-). We conclude that the HPLC/fluorescence assay using HE as a probe is more suitable [corrected] for detecting intracellular O(2)(.-).     

Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice

Non-invasive techniques to monitor stress hormones in small animals like mice offer several advantages and are highly demanded in laboratory as well as in field research. Since knowledge about the species-specific metabolism and excretion of glucocorticoids is essential to develop such a technique, we conducted radiometabolism studies in mice (Mus musculus f. domesticus, strain C57BL/6J). Each mouse was injected intraperitoneally with 740 kBq of 3H-labelled corticosterone and all voided urine and fecal samples were collected for five days. In a first experiment 16 animals (eight of each sex) received the injection at 9 a.m., while eight mice (four of each sex) were injected at 9 p.m. in a second experiment. In both experiments radioactive metabolites were recovered predominantly in the feces, although males excreted significantly higher proportions via the feces (about 73%) than females (about 53%). Peak radioactivity in the urine was detected within about 2h after injection, while in the feces peak concentrations were observed later (depending on the time of injection: about 10h postinjection in experiment 1 and about 4h postinjection in experiment 2, thus proving an effect of the time of day). The number and relative abundance of fecal [3H]corticosterone metabolites was determined by high performance liquid chromatography (HPLC). The HPLC separations revealed that corticosterone was extensively metabolized mainly to more polar substances. Regarding the types of metabolites formed, significant differences were found between males and females, but not between the experiments. Additionally, the immunoreactivity of these metabolites was assessed by screening the HPLC fractions with four enzyme immunoassays (EIA). However, only a newly established EIA for 5alpha-pregnane-3beta,11beta,21-triol-20-one (measuring corticosterone metabolites with a 5alpha-3beta,11beta-diol structure) detected several peaks of radioactive metabolites with high intensity in both sexes, while the other EIAs showed only minor immunoreactivity. Thus, our study for the first time provides substantial information about metabolism and excretion of corticosterone in urine and feces of mice and is the first demonstrating a significant impact of the animals' sex and the time of day. Based on these data it should be possible to monitor adrenocortical activity non-invasively in this species by measuring fecal corticosterone metabolites with the newly developed EIA. Since mice are extensively used in research world-wide, this could open new perspectives in various fields from ecology to behavioral endocrinology.     

大鼠骨髓间充质干细胞的分离培养和外源基因的导入

目的探讨绿色荧光蛋白基因转染骨髓间质干细胞的可行性。方法采用F icoll-PaqueTMP lus淋巴细胞分离液,根据细胞密度梯度原理,分离大鼠骨髓间充质干细胞(rM SC s)并进行体外原代培养和传代扩增,倒置相差显微镜观察细胞生长情况,免疫细胞化学法对其初步鉴定。流式细胞仪分析转染效率。结果原代和传代培养的细胞呈现梭形外观,具有较强的生长增殖能力;细胞均一表达CD44、CD54、CD106、CD29抗原。电穿孔法转染rM SC s转染率为32.8%±3%。结论采用比重为1.077 g/L的F icoll-PaqueTMP lus能分离获得大鼠骨髓间充质干细胞,经原代培养和传代培养能够迅速扩增。电穿孔法具有较高的介导外源基因表达于rM SC s的效率。