宫颈癌组织中HPV16 E7序列多态性分析

目的探讨广东地区宫颈癌组织中HPV16肿瘤相关性抗原E7基因序列的多态性.方法采用通用引物PCR直接测序法对宫颈癌标本中的HPV分型,从含有HPV16型的标本中采用自行设计的多重引物通过巢式PCR扩增出HPV16E7,经DNA序列测定法检测其基因变异,进而寻找其热点突变.结果50例宫颈癌组织HPV-DNA的检出率为78%,其中HPV16和HPV18型混合感染18例,单纯HPV16型感染15例.33例含有HPV16型的标本中扩增出25例HPV16E7,其中17例647位核苷酸“T”变异“C”,导致相应的蛋白质由天冬氨酸变异为丝氨酸.结论广东地区宫颈癌组织中HPV16E7DNA序列发生碱基替换的区域主要在647位至846位,热点突变点为Nt647和Nt846.     

Concentration-dependent metabolism of diazepam in mouse liver

Previous mouse liver studies with diazepam (DZ),N-desmethyldiazepam (NZ), and temazepam (TZ) confirmed that under first-order conditions, DZ formed NZ and TZ in parallel. Oxazepam (OZ) was generatedvia NZ and not TZ despite that preformed NZ and TZ were both capable of forming OZ. In the present studies, the concentration-dependent sequential metabolism of DZ was studied in perfused mouse livers and microsomes, with the aim of distinguishing the relative importance of NZ and TZ as precusors of OZ. In microsomal studies, theK _ms andV _maxs, corrected for binding to microsomal proteins, were 34 μM and 3.6 nmole/min per mg and 239 μM and 18 nmole/min per mg, respectively, forN-demthylation andC ₃-hydroxylation of DZ. TheK _ms andV _maxs forN-demethylation andC ₃-hydroxylation of TZ and NZ, respectively, to form OZ, were 58 μM and 2.5 nmole/min per mg and 311 μM and 2 nmole/min per mg, respectively. The constants suggest that at low DZ concentrations, NZ formation predominates and is a major source of OZ, whereas at higher DZ concentrations, TZ is the important source of OZ. In livers perfused with DZ at input concentrations of 13 to 35 μM, the extraction ratio of DZ (E{DZ}) decreased from 0.83 to 0.60. NZ was the major metabolite formed although its appearance was less than proportionate with increasing DZ input concentration. By contrast, the formation of TZ increased disporportionately with increasing DZ concentration, whereas that for OZ decreased and paralleled the behavior of NZ. Computer simulations based on a tubular flow model and thein vitro enzymatic parameters provided a poorin vitro-organ correlation. TheE{DZ}, appearance rates of the metabolites, and the extraction ratio of formed NZ (E{NZ, DZ}) were poorly predicted; TZ was incorrectly identified as the major precursor of OZ. Simulations with optimized parameters imporved the correlations and identified NZ as the major contributor of OZ. Saturation of DZN-demethylation at higher DZ concentrations increased the role of TZ in the formation of OZ. The poor aqueous solubility (limiting the concentration range of substrates usedin vitro), avid tissue binding and the coupling of enzymatic reactions in liver, favoring sequential metabolism, are possible explanations for the poorin vitro-organ correlation. This work emphasizes the complexity of the hepatic intracellular milieu for drug metabolism and the need for additional modeling efforts to adequately describe metabolite kinetics. This work was supported by the Medical Research Council of Canada (MA-9104).     

原发性高血压患者冠状动脉粥样硬化与主动脉中央脉压的关系

背景:脉压和主动脉僵硬度都是冠心病的预测因子,主动脉脉压与动脉粥样硬化之间是一种互为因果的关系,对经冠状动脉造影确诊的冠心病与主动脉脉压的关系进行观察。目的:观察冠状动脉粥样硬化与原发性高血压患者中央脉压的关系。设计:病例-对照观察。单位:首都医科大学宣武医院心内科。对象:选择2002-07/2005-01到首都医科大学宣武医院心内科就诊的300例原发性高血压患者,均为初发或未经正规治疗,均符合国际高血压协会的诊断标准,并除外继发性高血压、心肌病、心脏瓣膜病、心力衰竭、肝肾功能不全等。其中男170例,女130例,平均年龄(61±11)岁。所有患者均对检测项目知情同意。方法:300例原发性高血压患者入院后均给予冠状动脉造影检查,根据是否患有冠心病将纳入患者分为冠心病组(共143例,其中男性92例,占64%)和非冠心病组(共157例,其中男性82例,占52%)。所有患者均签署知情同意。收集的参数包括:由侵入性方法测得的主动脉内收缩压和舒张压,冠状动脉的病变程度以及患者的基础临床资料。所有观察指标均以x±s表示,两组之间的比较采用独立样本的t检验,P<0.05认为有显著性差异。主要观察指标:两组患者主动脉收缩压、舒张压和脉压,反映动脉僵硬度的每搏输出量与主动脉脉压的比值;冠状动脉病变的支数及狭窄程度(狭窄程度<50%的病例归入非冠心病组);患者入院后的基础临床指标和常规生化指标(包括空腹血糖、血清肌酐、血脂指标等)。结果:纳入300例高血压患者全部进入结果分析,冠心病组的主动脉收缩压和脉压明显高于非冠心病组[(150.3±26.5),(145.6±23.3)mmHg,P<0.05]和[(77.1±22.7),(70.4±19.3)mmHg,P<0.05]。冠心病组主动脉脉压与每搏输出量的比值明显高于非冠心病组(1.20±0.44,0.96±0.33,P<0.05)。冠心病组患者的空腹血糖为(1.38±0.27)mmol/L,明显高于非冠心病组(1.08±0.28mmol/L,P<0.01),冠心病组患者的血清肌酐为(11.98±2.15)μmol/L,明显高于非冠心病组(11.19±1.58μmol/L,P<0.01),而高密度脂蛋白胆固醇明显低于非冠心病组[(0.54±0.13),(0.62±0.18)mmol/L,P<0.01]。结论:动脉粥样硬化可能与原发性高血压患者的主动脉僵硬度加重有一定关系,中央脉压可能因此升高。另外,动脉粥样硬化还将损害原发性高血压患者的肾功能,并影响脂质代谢。     

医疗纠纷处理应重视的几个问题

通过对医疗纠纷产生的原因进行分类,提出一套有针对性的处理医疗纠纷的方法,依照客观公正的原则及时处理,并建议医院成立一支处理医疗纠纷的的专门队伍和机构。     

HLA-B27相关的前葡萄膜炎眼后节并发症的临床特点

目的探讨HLA—B27阳性的前葡萄膜炎眼后节并发症的临床特点及治疗。方法2002年1月至2005年10月就诊的前葡萄膜炎患者采用流式细胞术进行外周HLA-B27的检测，筛选出71-例82眼HLA-B27阳性前葡萄膜炎患者，分为两组：实验组为眼后节受累的患者24例31眼，对照组为眼后节未受累的患者47例51眼。另有HLA—B27阴性前葡萄膜炎患者74例116眼作参考。所有患者均进行裂隙灯、眼底镜等常规眼科查体，部分行眼底荧光血管造影。分析了HLA—B27相关的前葡萄膜炎眼后节并发症的临床特点。采用x^2检验及t检验进行统计学分析。结果71例82眼HLA—B27阳性前葡萄膜炎中有24例31眼占37．8％出现眼后节并发症者，包括：黄斑囊样水肿12例15眼占48．4％，视乳头水肿7例8眼占25．8％，而玻璃体炎最多见为21例27眼占87．1％，8例10眼占32．3％患者存在两种或两种以上的眼后节表现。74例116眼HLA—B27阴性者有8例12眼占10．3％有眼后节表现。实验组中发生前房成型纤维素渗出31眼中18眼占58．06％和积脓31眼中9眼占29．03％明显高于对照组（前房纤维素渗出51眼中3眼，5．88％，X^2=27．56，P〈0．01；前房积脓51眼中1眼，1．96％，X^2=13．20，P〈0．01）；实验组中伴发相关全身性疾病24例中有16例占66．67％的几率高于对照组（47例中有17例，31．66％，X^2=5．94，P〈0．05）。出现眼后节并发症的患者经全身及局部应用糖皮质激素治疗，31眼中有11眼最佳矫正视力≥1．0。结论HLA-B27阳性前葡萄膜炎易出现眼后节并发症，HLA—B27相关的前葡萄膜炎眼后节受累的患者发生相关全身性疾病和前房纤维素渗出与积脓的几率较高。     

西宁地区健康小儿窦房结功能测定的初步探讨

应用食道心房调博缩测定健康小儿窦房结功能，以了解西宁地区正常数值与平原地区进行对比。选择２７例健康小儿，将Ｆ６二级电极导管从鼻腔送入，定位地食道心电图最大正负双相Ｐ波处，采用苏州东方电子仪器厂生产的ＤＦ－３Ａ型心脏电生理治疗仪进行检查，从而作出诊断。     

环孢菌素A阻断人HL－60抗药性细胞于G_1期而诱导敏感细胞凋亡

进一步研究了抗三尖杉酯碱的ＨＬ－６０细胞（ＨＲ２０）抗细胞凋亡的机制及该抗性和抗药性的关系。结果表明，环孢菌素Ａ（ＣｓＡ）２０，１０μｇ·ｍｌ￣（－１）诱导ＨＬ－６０细胞发生凋亡，而阻断ＨＲ２０细胞于Ｇ＿１期，就不能诱导细胞发生凋亡。低浓度的ＣｓＡ明显增加柔红霉素在ＨＲ２０细胞内的积聚，其逆转抗药性作用与阻断细胞周期运行无关。ＣｓＡ１０μｇ·ｍｌ￣（－１）处理ＨＲ２０细胞，可引起５０ｋＤａ的蛋白质高度磷酸化。结果提示：环孢菌素Ａ阻断抗三尖杉酯碱的ＨＬ－６０细胞于Ｇ＿１期，而诱导敏感的ＨＬ－６０细胞发生凋亡，其阻断作用与抗药性无关     

Immunoglobulin production in vitro by peripheral blood lymphocytes in systemic lupus erythematosus: helper T cell defect and B cell hyperreactivity

Peripheral blood lymphocytes (PBL) from 16 patients with systemic lupus erythematosus (SLE) and 15 healthy control subjects were cultured and immunoglobulin (Ig) production in vitro was measured by immunofluorescent staining for intracytoplasmic Ig, a reverse haemolytic plaque assay to quantify cells secreting Ig and a solid-phase radioimmunoassay for Ig secreted into culture supernatants. Compared with normal PBL, lymphocytes from patients with SLE produced significantly fewer Ig-containing cells, Ig-secretion cells (ISC) and less Ig in supernatants in cultures stimulated by pokeweed mitogen (PWM). These differences were most pronounced during phases of disease activity. Culturing SLE PBL with a supernatant obtained from PWM-activated cultures of normal T lymphocytes partially restored their capacity to produce ISC. This observation suggests a helper T cell defect of SLE lymphocytes. In addition, PBL from patients with active SLE generated more ISC when cultured with Staphylococcus aureus bacteria (S aureus) than with PWM. S. aureus-stimulated cultures of SLE PBL also generated more ISC than PBL from normal individuals. The S. aureus response of SLE lymphocytes did not correlate with disease activity. As S. aureus is a T cell-independent mitogen, the latter observations suggest that in SLE an intrinsic B cell hyperreactivity may be a more persistent defect whereas T cell defects are transitory.     

Association between genetic variants in sortilin-related receptor 1 (SORL1) and Alzheimer's disease in adults with Down syndrome

Recent reports have suggested that variants in the sortilin-related receptor gene (SORL1) increase the risk of late onset Alzheimer's disease (AD) in Northern European, Hispanic, African–American and Isreali–Arab populations. SORL1 directs trafficking of amyloid precursor protein (APP) and under-expression of SORL1 may lead to over-expression of β amyloid peptides. Adults with Down syndrome (DS) over-express APP and have early onset and high risk for AD. We investigated the relation of seven variants in the gene for SORL1 to age at onset and risk for AD among 208 adults with DS, 45–70 years of age at baseline. Participants were ascertained through the New York State developmental disability service system and followed at 18-month intervals. Information from cognitive assessments, caregiver interviews, medical record review and neurological examination was used to establish the diagnosis of dementia. Homozygosity for the minor T allele in rs556349 and for the minor C allele in rs536360 was associated with later age at onset and reduced risk of AD (HR = 0.26, 95% CI: 0.08–0.86; and HR = 0.40, 95% CI: 0.16–0.98, respectively). Mean age at onset was approximately four years later in individuals who were homozygous for those alleles compared with those who had at least one major allele. These findings indicate a modest association of variants in SORL1 with AD. In addition, we did not observe the same alleles to be associated with AD compared with earlier studies, suggesting that these SNPs are in linkage disequilibrium (LD) with the putative functional variants or that expression of the SORL1 gene and hence its interaction with APP might be modified by the extremely high levels of APP characteristic of Down syndrome. Thus, further studies are needed to identify functional variants that influence risk for AD in this uniquely vulnerable population.