人MxA基因启动子-88/-123位多态性荧光PCR检测方法的建立及临床应用

目的 建立一套快速、灵敏和特异的人抗黏液蛋白A(MxA)基因启动子中干扰素刺激应答元件-88/-123位多态性检测体系,为合理应用IFN-α治疗慢性乙型肝炎提供分子生物学检测手段.方法 对经IFN-α治疗的患者进行HBV DNA、HBV基因分型及MxA基因启动子中干扰素刺激应答元件-88/-123位多态性位点检测,确定基因多态性与IFN-α治疗效果之间的关系.通过各种条件优化实验,建立MxA基因启动子中干扰素刺激应答元件-88/-123位多态性荧光PCR检测体系,并通过与DNA序列分析法检测结果的对比,验证荧光PCR检测体系的灵敏性及特异性,从而对该体系的临床适用性进行初步评估.采用卡方检验计算P值、回归分析法计算对比率和95%可信区间.结果 MxA基因启动子干扰素刺激应答元件中-88位为G/T杂合型和-123位为C/A杂合型者皆为IFN-α敏感型,-88位为G/G纯合型和-123位为C/C纯合型者为IFN-α不敏感型.与DNA序列分析的金标准相比,荧光PCR检测的符合率为99.65%.结论 荧光PCR检测体系,可灵敏、快捷地检测患者MxA基因多态性位点.     

Pseudomonas aeruginosa possesses homologues of mammalian phenylalanine hydroxylase and 4 alpha-carbinolamine dehydratase/DCoH as part of a three-component gene cluster.

Pseudomonas aeruginosa possesses a multigeneoperon that includes phenylalanine hydroxylase (PhhA; phenylalanine4-monooxygenase, EC 1.14.16.1). phhA encodes PhhA (M(r) = 30,288), phhB (M(r) =13,333) encodes a homologue of mammalian 4 alpha-carbinolaminedehydratase/homeodomain protein transregulator, and phhC encodes an aromaticaminotransferase (M(r) = 43,237). The reading frames specifying phhB and phhCoverlap by 2 bases. The P. aeruginosa PhhA appears to contain iron and is pterindependent. Unlike the multimeric mammalian hydroxylase, the native P. aeruginosaenzyme is a monomer. The P. aeruginosa PhhA is homologous with mammalian PhhA,tryptophan hydroxylase, and tyrosine hydroxylase. Expression of PhhA from itsnative promoter required phhB. This may suggest a positive regulatory role forphhB, consistent with the dual catalytic and regulatory roles of thecorresponding mammalian homologue.     

野生型p^53基因导入对培养的兔血管平滑肌细胞生长的抑制作用

为探索腺病毒载体重组野生型ｐ５３基因转染平滑肌细胞的效率及其应用于动脉粥样硬化和动脉再狭窄的基因治疗的可能性，构建了野生型ｐ５３基因的生长缺陷型重组腺病毒载体。在体外培养条件下转染兔血管平滑肌细胞。应用生化染色法、免疫组化法及聚合酶链技术等检测了外源基因的转染效率及表达效果。应用细胞计数及同位素参入技术测定了ｐ５３基因对平滑肌细胞的抑制效果。流式细胞仪检测了平滑肌细胞。结果显示：腺病毒载体可快速有效地将外源基因转导进入平滑肌细胞，在转染细胞内可检测到外源性ｐ５３ｃＤＮＡ，并可高效表达ｐ５３蛋白。ｐ５３重组腺病毒载体转染细胞后，可显著抑制细胞生长，细胞计数减少，ＤＮＡ合成减少，并可导致细胞死亡。流式细胞仪检测有部分细胞体积缩小，ＤＮＡ减少。结果提示，野生型ｐ５３基因重组腺病毒载体转染平滑肌细胞可有效抑制细胞生长，有可能成为动脉粥样硬化和再狭窄的基因治疗手段。     

DNA切除修复酶的表达及与肺癌预后的关系

目的　探讨DNA切除修复鼠缺陷交叉互补基因 2 (ERCC2 )蛋白、尿嘧啶DNA糖基化酶(UDG)、增殖细胞核抗原 (PCNA)在不同肺组织中的表达及与肺癌预后的关系。方法　免疫组化法测ERCC2、UDG、PCNA在良性病变、癌周组织、肿瘤组织中的表达水平 ,并与临床、病理资料进行Ridit分析。结果　肿瘤组织、癌周组织ERCC2、UDG、PCNA表达与良性病变组织相比 ,差异有显著性 (P值均<0 0 5 ) ,其中ERCC2、UDG表达低于后者 ,PCNA表达高于后者 (P <0 0 0 1) ;但癌周组织中UDG的差异未达显著界值 (P >0 0 5 )。肿瘤组织与癌周组织相比 ,ERCC2、UDG、PCNA表达差异无显著性 (P值均 >0 0 5 )。年龄、吸烟、肺癌组织分型、肿瘤转移对ERCC2、UDG、PCNA的表达无显著影响 (P值均 >0 0 5 ) ,但UDG在低分化癌和肿瘤 >3cm的表达水平显著降低 (P <0 0 5 ) ,而PCNA则相反 (P <0 0 5 )。结论　UDG、PCNA的表达水平与肺癌分化、大小有关 ,可作为肺癌预后的指标。     

静脉注射尼卡地平对重度高血压患者血压和心功能的影响

目的研究静脉注射尼卡地平对重度高血压患者血压和左心功能的影响。方法用尼卡地平持续２４ｈ静脉滴注（１．５～３ｍｇ／ｈ）治疗２０例重度高血压患者，观察给药前后血压、心率、左心功能以及自觉症状变化。结果降压显效率１００％，５ｍｉｎ起效，０．５～１ｈ血压降至理想水平，同时全部病例脑循环障碍症状消失，２４ｈ降压效果平稳，心率轻度增快（＜１０ｍｉｎ－１），左心收缩功能明显改善，而舒张功能未见变化。结论静脉注射尼卡地平可迅速、显著而平稳降低重度高血压患者的血压，改善左心收缩功能。     

Protective effects of recombinant human growth hormone on cirrhotic rats

AIM: To investigate the effects and molecular mechanisms of recombinant human growth hormone (rhGH) on protecting liver function and alleviating portal hypertension of liver cirrhotic rats. METHODS: Liver cirrhosis of male Sprague-Dawley rats was induced by administration of thioacetamide. The rats with or without liver cirrhosis were randomly divided into four groups. Group A consisted of the normal rats was treated with normal saline (NS), group B consisted of the normal rats was treated with rhGH, group C consisted of cirrhotic rats was treated with NS, and group D consisted of cirrhotic rats was treated with rhGH. The rats of different groups were subcutaneously injected with 0.5 mL of NS or 333 ng/kg of rhGH daily for 7 d. After treatments, the following parameters were examined, including GH-binding capacity (R(T)) by (125)I-hGH binding, growth hormone receptor mRNA(GHR mRNA) expression by RT-PCR, relative content of collagen (RCC) by histomorphomertry, and level of malon-dialdehyde (MDA) and superoxide dismutase (SOD) in liver tissue by thiobarbituric acid reaction and pyrogallic acid self-oxidation, respectively. Serum albumin (ALB), alanine transaminase (ALT) and portal vein pressure (PVP) were also examined. RESULTS: rhGH up-regulated both the GH-binding capacity (R(T)) and the expression of GHR mRNA in vivo. R(T) in group A (72+/-12 fmol/mg protein) was significantly higher than that in group C (31+/-4 fmol/mg protein) (P<0.05). R(T) in group B (80+/-9 fmol/mg protein) increased markedly compared to group A (P<0.05). R(T) in group D (40+/-7 fmol/mg protein) raised remarkably compared with group C (P<0.05), but less than that in group A, and there was no significant GH binding affinity contrast (Kd) change. The GHR mRNA level (iOD, pixel) in group A (29+/-3) was significantly higher than that in group C (23+/-3) (P<0.05). GHR mRNA levels were significantly raised in group B (56+/-4) and group D (42+/-8) compared with groups A and C (29+/-3 and 23+/-3, respectively) (P<0.05). Compared with the normal liver, MDA level was higher and SOD level was lower in cirrhotic livers. After rhGH treatment, MDA level was significantly declined to 12.0+/-2.2 nmol/mg protein and SOD was raised to 1 029+/-76 U/mg protein in group D (P<0.05). ALB levels in groups B and D (42+/-7 g/L and 37+/-7 g/L, respectively) were significantly raised compared with those in groups A and C (35+/-5 g/L and 29+/-4 g/L, respectively) (P<0.05). ALT level was markedly lower in group D (69+/-7 U/L) compared to group C (89+/-15 U/L) (P<0.05), and close to group A (61+/-10 U/L). RCC in group C (22.30+/-3.86%) was significantly higher than that in group A (1.14+/-0.21%) and group D (14.70+/-2.07%) (P<0.05). In addition, rhGH markedly alleviated portal hypertension in liver cirrhotic rats (group D vs C, 9.3+/-1.5 cmH(2)O vs 14.4+/-2.0 cmH(2)O) (P<0.05). CONCLUSION: Pharmacological doses of rhGH can increase R(T) and GHR mRNA expression, ameliorate liver functions, repress fibrosis and decline portal hypertension, suggesting it has potentially clinical usage as a hepatotropic factor.     

Tdentification of a new target region on the long arm of chromosome 7 in gastric carcinoma by loss of heterozygosity

AIM: To define the common deleted region on the long arm of haman chromosome 7q linked to primary gastric carcinomas in Chinese by loss of heterozygosity (LOH)and its clinical significance.METHODS: Nine microsatellite markers distributed over chromosome 7q with an average marker density of 10cM were used to examine 70 primary gastric carcinomas for LOH by PCR amplification. The PCR products were separated by electrophoresis on polyacrylamide gel.Genescan and Genotyper soft-wares were used to analyze LOH.RESULTS: LOH with at least one marker on 7q occurred in 34.3% (12/50) of the tumors. Among them, LOH at D7S486 and D7S798 was higher in 24.0% (24/70) and 19.2% (5/26), respectively. By statistical analysis we also observed an obvious genotype-phenotype correlation on 7q (P＜0.05). The frequency of LOH at D7S486 in patients with lymph node metastasis was significantly higher than that in those without lymph node metastasis (P= 0.015).CONCLUSION: The high incidence of LOH at D7S486and its correlation with poorer prognosis suggest that there might be putative tumor suppressor genes in this region involved in the tumorigenesis and progression of gastric carcinoma.     

心脏磁共振特征组织追踪成像定量评价健康人群左心室心肌形变

目的:探讨3.0T心脏磁共振(CMR)特征组织追踪成像(FT)测量中国汉族健康人群左心室心肌形变参数的可行性,并分析年龄、性别对各应变参数的影响。方法:本研究共纳入健康志愿者53例。其中男27例,女26例。随后每组纳入人群按照年龄分为青年、中年及老年3个亚组。所有纳入者均行3.0TCMR检查。分析、对比不同性别和不同年龄组人群左心室整体及局部的径向、周向及纵向峰值应变(PS)、峰值收缩期应变率(PSSR)、峰值舒张期应变率(PDSR)。结果:其中,女性整体周向PS、纵向PS、心基底段周向PS大于男性;男性整体周向PSSR及心中间段周向PSSR、纵向PSSR大于女性。分析不同年龄段心肌形变结果显示,整体径向PS在男女性中老年组均明显高于中年组和青年组;女性组整体周向PS、周向PDSR老年组低于青年组。左心室整体径向、周向、纵向PS分别与部分心功能参数相关。结论:3.0TCMR-FT能客观地量化健康人群左心室心肌形变的相关参数,且不同性别、不同年龄段心肌各形变参数也存在差异,制定相应正常人群心肌形变参数范围需考虑性别及年龄的影响。     

夏幼周教授防治小儿腹泻的经验

本文介绍了著名老中医夏幼周教授运用祖传秘方“儿泻宁”治疗小儿腹泻的临床经验，并附有１３８例病案资料。本组患几经用“儿泻宁“治疗后，痊愈１０６例，占７７％；好转２９例，占２１％；无效３例，占２％。总有效率为９８％。其治疗关键在于治疗不离脾胃，久泻宣补肾，食滞则攻补兼施。