肝炎基因诊断芯片对丙型肝炎病毒基因分型检测的初步研究

目的 研究丙型肝炎病毒（HCV）基因分型诊断芯片的制备和其对丙型肝炎患者血清HCVRNA基因分型诊断的准确性。方法 根据丙型肝炎病毒5′端非编码区末端（5UTR）和C区设计引物和特异性探针,将设计的20条特异性寡核苷酸探针用核苷酸合成仪合成后,配制成浓度为50μmol/ml的点样液,用点样仪点到特殊处理的玻片介质上,制成丙型肝炎病毒基因分型诊断芯片,收集HCVRNA阳性的丙型肝炎患者血清60份作为实验组,健康人血清60份作为对照组,用荧光定量PCR仪对两组血清进行HCVRNA定量检测,实验组血清HCVRNA含量均大于500拷贝/ml,对照组血清HCVRNA含量均小地500拷贝/ml,两组血清进行HCVRNA分离纯化,逆转录反应,巢式PCR扩增后,分别用基因芯片,HCVRNA核酸序列测定法（美国Li-Cor核酸序列测定仪）进行双盲丙型肝炎基因分型检测。结果 实验组血清基因诊断芯片检测均为阳性;基因分型;1b型46例,3a型3例。3b型3例,2a型2例,2b型2例,混合型1b 2a2例。1b 3b型2例;核酸序列测定分型1b型50例,3a型3例,3b型3例,2a型2例,2b型2例;基因诊断芯片检测和核酸序列测定检测的基因分型结果符合率为93.3%。对照组血清基因芯片检测匀阴性。结论 肝炎病毒基因诊断芯片可以用于检测血清HCVRNA,并进行基因诊断分型,准确率较率。     

DNA芯片技术诊断乙、丙型病毒性肝炎的研究

目的：研究病毒性肝炎基因芯片的制备,用肝炎基因诊断芯片,病毒抗原抗体及DNA/RNA（PCR）检测方法对照检测乙型和丙型肝炎血清,比较两种方法的优缺点。方法：血清来自乙型肝炎患者40例,丙型肝炎患者20例和健康人40例,乙型肝炎确诊用雅培设备试剂,荧光微离子法;抗HCV检测丙型肝炎及PCR法检测HCVRNA,确诊后用健康人血清一起进行双盲混合编组,用肝炎基因诊断芯片检测。结果：40例乙型肝炎病毒标志阳性血清,基因诊断芯片检测阳性30例,阴性10列;12例HCVRNA阳性血清,基因诊断芯片检测阳性8例;抗HCV阳性,HCVRNA阴性血清8例,基因诊断芯片检测阳性2例,阴性6例;40例健康人血清,乙型和丙型肝炎基因诊断芯片检测均为阴性。结论：肝炎基因诊断芯片可以同时检测乙型和丙型肝炎,诊断乙型肝炎的准确率可达80%,假阳性率低;诊断丙型肝炎的阳性率低,技术有待完善。     

微板核酸杂交技术对江西地区HCV基因分型的观察

目的 采用微板核酸杂交-ELISA技术对HCV RNA进行基因分型。方法 采用PCR、核酸杂交和酶联显色技术，首先将HCV RNA进行RT-PCR扩增，并将扩增产物加入预先包被对HCV通用探针的徽孔板，再加入HCV各基因型显色探针，同时进行微板核酸杂交-ELISA显色，对江西地区129例临床诊断为丙肝RNA阳性患者血清中的HCVRNA进行基因分型研究。结果 江西地区HCV基因型可分为1b、2a、2b、3a、3b、6a、la／2a、1b／2a、1b／3a、1b／6a、3a／5a、2a／3a共12种单一基因型和混合基因型，其中以1b基因型为主，占50．39％；慢性丙肝患者的基因型较为复杂，存在较高比例的混合基因型。结论 江西地区HCV感染者以1b基因型为主．PCR徽板核酸杂交-ELISA方法可以准确、快速、简便进行HCV基因分型，在探讨HCV的流行病学及观察药物对各基因型的疗效方面具有重大的意义。     

病毒性肝炎基因诊断芯片测定HBV DNA、HCV RNA的价值

目的：研究病毒性肝炎基因芯片的制备,评价该基因芯片对乙型和丙型肝炎的诊断价值。方法：收集乙型肝炎40例,丙型肝炎20例,40例健康人,乙型肝炎确诊用美国雅培设备试剂,荧光微离子法;丙型肝炎检测抗HCV及PCR法检测HCV RNA。确诊后同健康人血清一起进行双盲混合编组,用肝炎基因诊断芯片检测。结果：40例乙型肝炎病毒标志阳性血清,基因诊断芯片检测阳性30例,阴性10例;12例HCV RNA阳性血清,基因诊断芯片检测阳性8例;抗HCV阳性、HCV RNA阴性血清8例,基因诊断芯片检测阳性2例,阴性6例。40例健康人血清,乙型和丙型肝炎基因诊断芯片检测均为阴性。结论：肝炎基因诊断芯片可以同时检测乙型和丙型肝炎,诊断乙型肝炎的准确率可达80％,假阳性率低;诊断丙型肝炎的阳性率低,技术有待完善。     

天津地区丙型肝炎病毒基因型与临床的相关性研究

目的：研究丙型肝炎病毒（HCV）基因型,了解天津地区丙型肝炎的分布情况。方法：收集252例丙型肝炎患者的血清标本,采用基因测序法进行HCV基因型的检测,并对基因型与其他临床指标进行分析。结果：252例丙型肝炎患者分别感染1b、1a、1c、2a、2i、3a、3b、4a和6a共9种基因亚型,其中1b型占59．9％,其次是3a型,占12．7％。对1型、2型、3型和4型4种基因型进行肝功能、HCVRNA病毒载量比较,经单因素方差分析有明显统计学差异（P〈0．05）;对HCV感染的不同肝病程度患者的基因型分布进行分析,无显著性差异（P〉0．05）。结论：天津地区丙型肝炎病毒感染以1b型为主,其次为3a型。HCV基因型与HCVRNA病毒载量、肝脏损伤指标有明显统计学差异。     

南通地区慢性丙型肝炎患者的HCV基因分型

目的 :了解南通地区慢性丙型肝炎患者的HCV基因型分布情况。方法 :将 5 3例抗HCV阳性的血清标本用RT -PCR法检测HCV -RNA ,然后采用膜显色结果判读的基因芯片对 3 2例HCV -RNA阳性者进行基因分型研究。结果 :南通地区HCV感染检出 1b、1a、2a、1b + 1a、1b + 1a + 2a共 5种单一基因型和混合基因型 ,其中以 1b型为主 ,占 5 3 .1%。结论 :南通地区 1b型为优势株 ,2a和 1b + 1a型居次 ;1b + 1a型高于全国其他地区。     

基因芯片检测血清标本的丙型肝炎病毒基因型

目的：探讨丙型肝炎病毒(HCV)基因分型检测芯片在临床的初步应用及意义。方法：基因芯片检测150例血清标本中HCV基因型，并与HCVRNA定性及测序结果进行比较。结果：HCV基因分型检测芯片检出的阳性结果与HCV—RNA定性及测序结果的相对符合率为100．0％。结论：HCV基因分型检测芯片是一种准确性好、灵敏度高、特异性强、操作简便快速的基因检测方法，可以为临床诊断、治疗提供有力保证。     

血清中不同基因型丙型肝炎病毒RNA含量的测定

目的了解血清中不同基因型丙型肝炎病毒HCVRNA的含量。方法用实时荧光定量RTPCR和逆转录型特异性引物PCR同时检测21例费城酒精依赖丙肝患者的血清HCVRNA拷贝数并分析HCV基因型。结果21份血清标本中,有17份为HCVRNA阳性。这些阳性血清的HCVRNA含量波动范围在104.60～106.20拷贝/ml。检测到1a和1b两种HCV基因型,其中1a型7例,RNA平均滴度为105.28±0.75拷贝/ml,1b型(9例)RNA平均滴度为105.28±0.28拷贝/ml。结论定性和定量检测结果有极佳的一致性,酒精依赖丙肝基因型1a和1b血清中HCV含量无显著性差异。     

原发性肝细胞癌与HCV分型的关系

目的：了解原发性肝细胞癌（HCC）患者HCV基因型分布现状及其关系。方法：选择山西省肿瘤医院1996年间住院的HCVRNA阳性，临床上确诊的HCC患者血清标本13份，非肝癌癌症患者血清标本15份，1996年－1997年HCVRNA阳性的公共场所从业人员血清标本15份，用型特异性引物进行RT－PCR基因分型。结果：13例HCVRNA阳性的HCC患者，除1例未能分型外，其余12例均为HCV 1b型（100％）。15例非肝癌癌症患者，除13酌（86．67％）为1b型感染者外，其余均为2a型（13．33％）。15例从业人员中，不仅检出了12b型感染（86．67％），还检出2a型，1／2混合型各1例。所有标本中1b,2a型，1b/2a混合型的构成分别为38／42（90．48％），3／42（7．14％），1／42（2．38％），以1b型的比例最高，未检出1a,2b和3a型。结论：HCVRNA1b型与HCC的关系较为密切，同时也是山西地区HCV的感染优势株。     

联合分析HCV种属信息区和高度保守区序列确定广东地区HCV基因型和亚型

目的扩增HCV种属信息区NS5B和高度保守区5-UTR基因序列,用系统进化树法深入分析HCV基因型和亚型,阐明广东地区丙型肝炎患者HCV基因型和基因亚型的特点。方法采集99例慢性丙型肝炎病例血清,应用RT-PCR技术分别扩增HCVNS5B区和5-UTR特定片段,测序后进行系统进化树分析,确定HCV基因型及亚型。结果99例患者标本均准确分型,其中NS5B区段分型74例,5-UTR区段分型99例。HCV基因型和亚型的分布:1a亚型1例、1b亚型59例、1型7例、2a亚型11例、3a亚型3例、3b亚型2例、3型3例、6a亚型13例;另有2例标本在两区段分型不同,5-UTR/NS5B区段分型分别为6a/1b和3a/1b。两区段共同分型74例,72例标本在两区段基因型一致,2例标本在两区段基因型不同,为HCV重组体。结论联合分析HCV不同区段基因序列,可提高HCV基因分型的准确度及灵敏性。广东地区慢性丙型肝炎患者HCV基因亚型分布以1b为主;其次为6a和2a;3型中的a、b亚型也占有一定比例。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.