宫颈癌细胞中DAPK1基因CpG岛甲基化及其表达

目的 :探讨宫颈癌细胞中凋亡相关蛋白激酶 1(DAPK1 )基因CpG岛甲基化及其表达与宫颈癌的相关性 .方法 :应用甲基化特异性PCR和SABC免疫组化方法 ,检测 32(鳞癌 1 8,腺癌 1 4 )例宫颈癌组织DAPK1基因CpG岛甲基化修饰及蛋白表达 .结果 :宫颈癌中DAPK1基因甲基化扩增阳性率明显增高 5 6 .3% ,与正常宫颈比较有显著性差异 (P <0 .0 5 ) ,鳞癌与腺癌甲基化扩增阳性率无显著差异 (P >0 .0 5 ) ;宫颈癌中DAPK1蛋白表达阳性率降低 1 5 .6 3% ,与正常宫颈比较有显著性差异 (P <0 .0 5 ) ,鳞癌和 7例与腺癌无显著性差异 (P >0 .0 5 ) .结论 :DAPK1基因CpG岛异常甲基化修饰能抑制DAPK1的转录 ,使DAPK1蛋白不表达 ,丧失抑癌作用 ,是促进正常宫颈上皮细胞癌变的一个重要因素     

Effects of buffering in hypercapnia and hypercapnic hypoxemia

Anesthetized, paralyzed and mechanically ventilated pigs were exposed to extreme hypercapnia (Paco_2-20 kPa) at Fio₂ 0.4 for 480 min, with (n = 6) or without (n = 6) continuous infusion of isotonic buffers (bicarbonate and trometamol). Arterial pH was higher in buffered animals than controls, 7.21 ±0.01 vs 7.01±0.01 (mean ± s.e.mean, P < 0.01). Serum osmolality and Paco₂ did not differ between groups throughout the experiment. The hemodynamic response to hypercapnia was attenuated in the buffered group, who had lower heart rate, 133 ± 6 vs 189±12 min^-1 (P < 0.01), mean arterial pressure (MAP) 109 ± 4 vs 124 ± 4 mmHg (14.5 ± 0.5 vs 16.5 ± 0.5 kPa) (P < 0.05), mean pulmonary arterial pressure 16±1 vs 23 ± 1 mmHg (2.1 ±0.1 vs 3.1 ±0.1 kPa) (P < 0.01), and pulmonary vascular resistance (PVR) 249 ± 21 vs 343 ± 20 dyn s-cm^-5 (2490±210 vs 3430±200 μN-s-cm^-5) (P < 0.01), compared with the control group. Subsequently, both groups were exposed to hypercapnic hypoxemia by stepwise increases in Fio₂ (0.15, 0.10, 0.05) at 30-min intervals, while Fico₂ was kept at 0.2. PVR increased in both groups (P < 0.05) but, except for heart rate, all hemodynamic differences between the groups disappeared during hypoxia. At Fio₂ 0.15, buffered animals had higher arterial oxygen saturation (73 ± 5%) than the controls (55 ± 5%), (P < 0.05). The control animals died after 1–29 min (mean 14 min) at Fio₂ 0.10, while all buffered animals survived Fio₂ 0.10 with stable MAP (122 ± 14 mmHg (16.3 ± 1.9 kPa). The buffered animals died after 4–22 min (mean 15 min) at Fio₂ 0.05. We conclude that buffering to a pH of 7.21 attenuates the observed hemodynamic response in extreme hypercapnia and improves survival in hypercapnic hypoxemia.     

Concurrent oral and genital infection with HPV DNA types 6 and 11 in a heterosexual male

Objective Detection of HPV DNA in oral and genital lesions of a heterosexual male. 4 months after oral and vaginal intercourse with a woman with vulvar warts. Passible modes of acquisition of oral HPV infection in the male sexual partner are discussed. Setting Genitourinary Medicine clinic. Methods Polymerase chain reaction amplification of genomic DNA from oral and genital lesions. HPV DNA typing by dot blot hybridization. Results HPV DNA types 6 and 11 were identified in a polypoid tongue lesion and in a penile wart from the male sexual partner. Conclusions The acquisition of oral HPV infection in the male sexual partner may have resulted from genital-oral HPV transfer, either by direct contact with vulvar warts or by digital self-inoculation.     

特异性DNA倍增技术检测t-PA cDNA基因在表达细胞中的稳定性

利用特异性DNA倍增技术(polymerase chain Reaction,PCR)检测t-PA cDNA基因在表达细胞基因组中的稳定性,并对所得的PCR反应产物进行了限制性内切酶片段、分子杂交和核苷酸顺序分析等方面的研究,证实了t-PA cDNA基因已插入到表达细胞染色体中。这种方法快速、简便、灵敏度和特异性高,是检测基因工程表达细胞中cDNA基因稳定整合状况的好方法。     

A newly recognized point mutation in the cytochrome b558 heavy chain gene replacing alanine57 by glutamic acid,in a patient with cytochrome b positive X-linked chronic granulomatous disease

Molecular genetic analysis was performed in a patient with cytochrome b positive X-linked chronic granulomatous disease. A previous Southern blot study, using a cytochrome b heavy chain cDNA as probe, revealed a Pst I restriction fragment pattern for the cytochrome b heavy chain gene (CYBB) different to that of normal individuals. Since restriction length polymorphism with Pst I has never been observed in control individuals and no abnormal restriction fragment patterns in the patient's CYBB was detected with seven other enzymes used, we focussed on the single Pst I site in the CYBB cDNA as being the only mutation site responsible for his disease. A fragment of the patient's cDNA which included the Pst I site was amplified by reverse polymerase chain reaction, and loss of the Pst I site in the fragment was confirmed by incubation with Pst I. Subsequent sequence analysis of the fragment revealed a point mutation in the Pst I site (cytosine to adenine), substituting glutamic acid for alanine at position 57.     

SRY基因诊断在临床中的应用

应用聚合酶链式反应（ＰＣＲ）扩增技术，对１０例染色体核型为４６，ＸＹ和１例为４６，ＸＸ／４６，ＸＹ的性反转、睾丸女性化、两性畸形及男性生殖器发育不良的患者进行了性别决定区（ＳＲＹ）基因检测。结果表明９例４６，ＸＹ和１例４６，ＸＸ／４６，ＸＹ患者的ＳＲＹ基因存在，１例４６，ＸＹ女性患者的ＳＲＹ基因缺失。该结果表明能够用分子生物学技术对性反转、性发育异常的病因进行分析     

膀胱肿瘤组织中端粒酶催化亚基和c-myc的表达

目的 研究膀胱肿瘤组织中端粒酶催化亚基(hTERT)和c-myc间的表达及关系。方法 采用逆转录-聚合酶链反应(RT-PCR)法对27例膀胱肿瘤组织、16例癌旁组织和16例正常膀胱粘膜同时测定hTERT和c-myc表达,分析两指标在3种组织的表达差异和相关性。结果 TCC组织hTERT和c-myc的表达阳性率分别为100.0％(27/27)和66.7％(18/27);癌旁组织两指标的阳性率均为 37.5％,而正常膀胱粘膜分别为 0和 25.0％;hTERT和c-myc两指标在膀胱肿瘤组织中的阳性表达有相关性(P<0.01)。结论 hTERT较c-myc作为TCC的诊断指标更灵敏和特异;两指标在TCC中的阳性表达具有相关性。