HPV18E2蛋白CTL表位多肽的构建与表达

目的分析HPV18E2蛋白的CTL表位,合成并纯化这些表位多肽,构建表达HPV18E2蛋白的靶细胞,检验特异性CTL对靶细胞的杀伤效果,为进一步研制人乳头瘤病毒18型（HPV18）基因工程疫苗打下基础。方法以重组质粒（pBR322-HPV18）为模板,利用PCR方法扩增HPV18E2 DNA片段,将HPV18E2 DNA与加强绿色荧光质粒（pIRES2-EGFP）重组构建重组质粒（pIRES2-HPV18E2-EGFP）。用酶切电泳及测序检查质粒重组后序列正确性。重组质粒转染Hela细胞。51^Cr释放实验评估CTL杀伤能力。结果克隆重组质粒pIRES2-HPV18E2-EGFP酶切后显示的酶切图谱与预期相同,而且测序验证插入片段全序列无改变。转染并用G418筛选后,在荧光显微镜下可见绿色荧光细胞的表达。三条候选多肽特异性杀伤能力分别为60.00%、71.29%和80.00%。结论三条备选肽段都具有明显的杀伤效应,均为HPV18E2蛋白的有效CTL表位。     

蛋氨酸合酶基因A275 6G位点多态性与非综合征型唇腭裂的关联性研究

Objective To study the association of the A2756G polymorphism of the methionine synthase (MS) gene with nonsyndromie cleft lip with or without cleft palate (NSCL/P) in Chinese. Methods Ninety-seven NSCL/P case-parent triads were selected as the case group. One hundred and four healthy subjects and their biological parents were selected as control group. For all subjects the A2756G polymorphism of the MS gene was examined by PCR-RFLP method. Results There was no statistical difference in genotype and allele frequencies for MS A2756G variants among family members between case group and control group. The GG genotype was not detected in the offsprings and mothers. The odds ratio and confidence interval of genotype AG in offspring, father and mother were 1.78(0.74-4.34), 0.80(0. 36-1.79) and 1.26(0. 54-2.93) respectively. The odds ratio and confidence interval of allele G in offspring, father and mother were 1.70(0.78-3.73), 0. 88(0. 49-1. 75) , and 1.23(0.59-2.60) respectively. The G allele did not increase the risk of NSCL/P. Transmission disequilibrium test (TDT) analysis yielded no evidence of linkage disequilibrium (χ2=0.034,P>0. 05). The results of haplotype-based haplotype relative risk (HHRR) analysis (χ2=0.03,P>0.05) and family-based association tests (FBAT) (Z=0. 186, P> 0.05) failed to show association between the MS A2756G variant and the risk of NSCL/P. Conclusion The A2756G polymorphism of the MS gene was not associated with NSCL/P in Chinese in the present study.     

蛋氨酸合酶基因A275 6G位点多态性与非综合征型唇腭裂的关联性研究

Objective To study the association of the A2756G polymorphism of the methionine synthase (MS) gene with nonsyndromie cleft lip with or without cleft palate (NSCL/P) in Chinese. Methods Ninety-seven NSCL/P case-parent triads were selected as the case group. One hundred and four healthy subjects and their biological parents were selected as control group. For all subjects the A2756G polymorphism of the MS gene was examined by PCR-RFLP method. Results There was no statistical difference in genotype and allele frequencies for MS A2756G variants among family members between case group and control group. The GG genotype was not detected in the offsprings and mothers. The odds ratio and confidence interval of genotype AG in offspring, father and mother were 1.78(0.74-4.34), 0.80(0. 36-1.79) and 1.26(0. 54-2.93) respectively. The odds ratio and confidence interval of allele G in offspring, father and mother were 1.70(0.78-3.73), 0. 88(0. 49-1. 75) , and 1.23(0.59-2.60) respectively. The G allele did not increase the risk of NSCL/P. Transmission disequilibrium test (TDT) analysis yielded no evidence of linkage disequilibrium (χ2=0.034,P>0. 05). The results of haplotype-based haplotype relative risk (HHRR) analysis (χ2=0.03,P>0.05) and family-based association tests (FBAT) (Z=0. 186, P> 0.05) failed to show association between the MS A2756G variant and the risk of NSCL/P. Conclusion The A2756G polymorphism of the MS gene was not associated with NSCL/P in Chinese in the present study.     

促肝细胞生长素治疗慢性重型肝炎疗效观察

为观察促肝细胞生长素 (PHGF)治疗慢性重型肝炎的疗效 ,对 2 0 0 0年 1月～ 2 0 0 2年 3月入院的 62例慢性重型肝炎患者以综合治疗和在综合治疗基础上加 PHGF方法进行分组观察 ,报告如下 :1　资料与方法1.1　一般资料　 62例慢性重型肝炎 ,诊断均符合 1995年全国传染病与寄生虫病学术会议修订的病毒性肝炎诊断标准 ,随机分为治疗组和对照组。治疗组 3 3例 ,男 3 0例 ,女 3例 ,年龄 11岁～ 43岁。对照组 2 9例 ,男 2 7例 ,女 2例 ,年龄 12岁～42岁。两组病例入院时谷丙转氨酶、血清胆红素、血清白蛋白、凝血酶原时间等指标统计学无显著性…     

pIRES2-HPV18E2-EGFP质粒重组及其在HeLa细胞中的表达

目的构建人乳头瘤病毒18型E2基因片段(HPV18E2)重组表达质粒并予以表达,为进一步研制HPV18相关疾病提供材料.方法以重组质粒(pBR322-HPV18)为模板,PCR方法扩增HPV18E2DNA片段,将HPV18E2DNA与加强绿色荧光质粒(pIRES2-EGFP)重组构建重组质粒(pIRES2-HPV18E2- EGFP).用酶切电泳及测序检查质粒重组后序列正确性.重组质粒转染Hela细胞.RT-PCR鉴定转染细胞中E2基因.结果 PCR扩增DNA片段约为1 kb,与预期结果相同.克隆重组质粒pIRES2-HPV18E2- EGFP酶切后显示的酶切图谱与预期相同,而且测序验证插入片段全序列无改变.转染并用G418筛选后,在荧光显微镜下可见绿色荧光细胞的表达.转染细胞RT-PCR出现特异性条带.结论成功构建表达HPV18 E2蛋白的靶细胞模型.     

蛋氨酸合酶基因A275 6G位点多态性与非综合征型唇腭裂的关联性研究

Objective To study the association of the A2756G polymorphism of the methionine synthase (MS) gene with nonsyndromie cleft lip with or without cleft palate (NSCL/P) in Chinese. Methods Ninety-seven NSCL/P case-parent triads were selected as the case group. One hundred and four healthy subjects and their biological parents were selected as control group. For all subjects the A2756G polymorphism of the MS gene was examined by PCR-RFLP method. Results There was no statistical difference in genotype and allele frequencies for MS A2756G variants among family members between case group and control group. The GG genotype was not detected in the offsprings and mothers. The odds ratio and confidence interval of genotype AG in offspring, father and mother were 1.78(0.74-4.34), 0.80(0. 36-1.79) and 1.26(0. 54-2.93) respectively. The odds ratio and confidence interval of allele G in offspring, father and mother were 1.70(0.78-3.73), 0. 88(0. 49-1. 75) , and 1.23(0.59-2.60) respectively. The G allele did not increase the risk of NSCL/P. Transmission disequilibrium test (TDT) analysis yielded no evidence of linkage disequilibrium (χ2=0.034,P>0. 05). The results of haplotype-based haplotype relative risk (HHRR) analysis (χ2=0.03,P>0.05) and family-based association tests (FBAT) (Z=0. 186, P> 0.05) failed to show association between the MS A2756G variant and the risk of NSCL/P. Conclusion The A2756G polymorphism of the MS gene was not associated with NSCL/P in Chinese in the present study.     

上皮型钙黏蛋白在宫颈癌中作用研究进展

上皮型钙黏蛋白（E-cadhenn，E-CD）是一种钙依赖性细胞黏附分子，分布于人和动物的上皮细胞，是维护上皮细胞形态和结构的完整性和极性的重要分子。近年来大量研究表明，E-CD及其复合物与宫颈癌的发生发展密切相关，对近年来E-CD在宫颈癌方面的研究作一综述。     

固化磷酸钙加钢板治疗肱骨近端粉碎性骨折

目的 讨论固化磷酸钙填塞和三叶钢板固定术治疗肱骨近端粉碎性骨折的疗效。方法 回顾分析2002年1月至2004年12月收治的11例肱骨近端粉碎性骨折临床资料，经肩关节前入路进入复位，三叶草状钢板螺丝钉内固定，并在断端骨折缺失部填入金世植骨灵，术后吊带悬吊，3周后功能锻炼。结果 11例均获随访（平均8．3个月），肩关节外展〉90°，能生活自理，X线片示无肱骨头坏死，骨折对位对线好，按Neer功能评分标准，优9例，良2例。结论 三叶板固定和固化磷酸钙植骨，能够保持骨断端的连续性，维持断端的稳定，对于伴有严重骨质疏松的老年患者，避免人工关节置换。     

自拟活血祛瘀散外敷治疗急性腕关节扭挫伤

腕关节损伤除包括组成腕关节的尺桡骨下端与8块腕骨及5个掌骨基部诸骨构成骨的骨折外；还包括连接以上骨组织并构成关节的外来韧带与内在韧带，尺腕三角软骨盘的急慢性扭、挫、撕裂伤。对于急性外伤后的腕部疼痛，活动受限，无力、肿胀等，即外伤性腕关节损伤，在排除腕关节骨折后，主要指外伤性参与腕关节构成的韧带、软骨的急性损伤。我科自2002年1月到2005年3月应用自拟活血祛瘀散外敷治疗急性腕关节扭挫伤110例，取得满意疗效，现报告于下。     

新生儿永存动脉干1例

患儿 ,男 ,57ｈ ,因“全身皮肤发灰 ,呻吟样呼吸2 0ｍｉｎ”由产科转入。系第 3胎第 1产 ,孕 4 1+2 周 ,因“胎儿宫内窘迫”剖宫产 ,出生体重 2 950 ｇ ,Ａｐｇａｒ评分 1分钟及 5分钟均为 10分 ,羊水Ⅲ°污染。查体 :Ｔ 36℃ ,Ｒ 80次 /ｍｉｎ ,刺激无反应 ,全身皮肤苍白发灰、下腹部散在数个出血点 ,指压皮肤充盈 >3ｓ,前囟平坦 ,双目凝视 ,瞳孔等大等圆 ,对光反应迟钝。唇绀 ,鼻扇 ,两肺呼吸音粗 ,心率 140次 /ｍｉｎ ,心音有力、无杂音。肝右肋下 4ｃｍ ,质中 ,脾未及 ,四肢瘫软 ,肢端厥冷 ,原始反应消失。辅检 :血ＷＢＣ12 .3× 10 9/…