单管半套式PCR-SSCP用于结核分支杆菌rpoB基因突变的研究

目的 ①证明单管半套式聚合酶链反应(PCR)是检测结核分支杆菌的特异性方法;②研究结核分支杆菌rpoB基因突变与利福平耐药性的关系。方法 设计三条针对结核分支杆菌的特异性引物,采用单管半套式PCR-单链构象多态性(SSCP)方法对结核分支杆菌、其它分支杆菌和富含GC碱基的细菌进行分析。结果 ①结核分支杆菌标准菌株H_(37)Rv和55株临床分离株均扩增出一条193bp片段,其特异性为100％,敏感性约为10pgDNA;15株非结核分支杆菌和8株富含GC的细菌均未得到扩增;②55株结核分支杆菌临床分离株193bp扩增片段SSCP图谱特点:以H_(37)Rv为对照,15株利福平敏感株均无区别;40株耐药株除6株外其余34株SSCP图谱有明显区别,检测阳性率为85％。结论 单管半套式PCR检测结核分支杆菌简便、敏感、特异,结合SSCP可检出耐利福平结核分枝杆菌rpoB基因突变,此突变与利福平耐药关系密切。     

结核分支杆菌耐药基因的杂交检测

目的采用PCR-反向斑点杂交检测结核分支杆菌中rpoB基因、KatG基因和rpsL基因突变。评价该方法检测结核分支杆菌利福平（RFP）、异烟肼（INH）和链霉素耐药性,探讨直接检测结核患者痰标本的可行性。方法用PCR-反向斑点杂交技术分析对75株临床分离株和45例肺结核患者涂阳痰标本进行rpoB、rpsL和KatG基因突变的检测。结果临床耐药株分离株进行rpoB、rpsL和KatG基因突变的检测,其突变率分别为83.9%、62.5%、57.1%。肺结核患者涂阳痰标本中耐药株突变率分别为81.0%、60.0%、52.9%。结论rpoB基因、KatG基因和rpsL基因突变与结核分支杆菌耐利福平、异烟肼和链霉素密切相关,应用PCR-反向斑点杂交技术可快速检测结核分支杆菌对异烟肼、链霉素和利福平耐药性。PCR-反向斑点杂交技术有望成为结核分支杆菌耐药性检测的重要方法之一。     

快速检测结核分支杆菌对利福平的耐药性

目的：建立快速检测结核分支杆菌耐药基因型的分子药敏试验方法。方法：用聚合酶链反应-单链构象多态性(PCR-SSCP)分析结核分支杆菌rpoB基因突变并与药敏试验对照。结果：116株结核分支杆菌的PCR-SSCP图谱与H37Rv为对照，81株耐药菌73株有明显区别，而35株敏感，仅1株位置有区别；检测阳性率90．1％；特异性97．1％。结论：rpoB基因突变是结核分支杆菌耐利福平的主要机理，PCR-SSCP可简便快速地检出rpoB基因的突变，有助于结核分支杆菌耐药性的快速检测和研究。     

基因芯片检测结核分枝杆菌对利福平和异烟肼耐药性

目的研究基因芯片在检测结核分枝杆菌对利福平和异烟肼耐药性中的应用。方法根据与利福平和异烟肼耐药相关的4条基因上的11个位点和30种单核苷酸多态性设计制作芯片,用芯片检测结核分枝杆菌的基因突变,从而判断其耐药性。结果用基因芯片在110株异烟肼耐药株中检测出85侏（73．3％）,在30株异烟肼敏感株中检测出22株（73．3％）,在94株利福平耐药株中检测出77株（81．9％）,在46株利福平敏感株中检测出40株（87．0％）。芯片检测结果与部分样本的测序结果完全相符。结论用DNA芯片检测结核分枝杆菌对利福平和异烟肼的耐药性具有较高的特异性和敏感性,该法快速、精确,可以应用于结核分枝杆菌耐药性的检测。     

PCR-SSCP技术直接快速检测痰标本中结核分支杆菌rpoB,katG,ahpC基因突变的研究

目的：应用PCR－SSCP技术直接快速检测痰标本中结核分支杆菌rpoB,katG及ahpC基因突变,评价此方法用于结核分杆菌利福平（RFP）及异烟肼（INH）耐药性试验的意义。方法：以结核分支杆菌H37Rv为对照,30例耐RFP（单耐或多耐）、21例耐INH（单耐和多耐）的结核分枝杆菌临床分离株及10例敏感菌株;39例菌阳肺结核患者及30例非结核 性肺部疾病患者痰标本,采用PCR－SS－CP技术检测痰标本和菌株中的结核杆菌rpoB,katG,ahpC基因突变,并与药敏试验对照。结果：PCR－SSCP检测痰标本中结核分支杆菌rpoB,katG及ahpC基因突变的阳性率分别为79％（31／39）、46％（18／39）及5％（2／39）.结论：PCR－SSCP可望成为直接检测临床痰标本中结核分枝杆菌耐利福平及耐异烟肼的快速方法。     

非结核分支杆菌16例药敏特点分析

目的:分析我院2008-2010年非结核分支杆菌(NTM)的耐药情况.方法:对2008年3月～2010年3月无锡市传染病医院结核病实验室所做结核分支杆菌药敏试验的279例菌株用TCH和PNB进行菌型初步鉴定,采用绝对浓度法对12种抗结核药物:异烟肼、利福平、链霉素、乙胺丁醇、吡嗪酰胺、奥氟星、对氨基水杨酸、卷曲霉素、力克肺疾、利福喷丁、丁胺卡那和丙硫异烟胺进行药敏试验,并对分离出来的16株NTM的药敏结果进行分析.结果:279例菌株中NTM分离率为5.73%,有3例全耐药;16株NTM对吡嗪酰胺呈现全耐药;除对丁胺卡那的低浓度与高浓度耐药率差异有统计学意义外,对其他10种药物低、高浓度的耐药率差异无统计学意义.说明很多非结核分支杆菌对抗结核药物呈天然的抗药性.结论:我市非结核分支杆菌处于较低的流行水平,针对非结核分支杆菌的高耐药性.需要制定新的、更有效的治疗手段,对NTM感染和NTM病应予高度重视.     

结核分支杆菌耐药基因检测结果分析

目的 分析肺结核患者结核分支杆菌获得性耐药情况,比较两种耐药性检测方法的检测效果.方法 用药敏试验(绝对浓度法)检测结核分支杆菌株对利福平(RFP)、异烟肼(INH)、链霉素(SM)、吡嗪酰胺(PZA)和乙胺丁醇(EMB)的耐药情况,采用聚合酶链反应-单链构象多态性分析(PCR-SSCP)检测结核分支杆菌耐药rpoB、katG、rpsL、pncA和embB基因突变.结果:400例肺结核耐药患者常规药敏试验耐药316例(79.8%);耐药基因检测耐药株288株,突变率72.0%.RFP耐药例数和耐药率明显高于SM、PZA和EMB(耐药例数:F=2.45,2.56,2.69,P＜0.05;耐药率:F=2.55,2.66,2.79,P＜0.05),rpoB突变株和突变率明显高于katG、rpsL、pncA和embB(突变株:F=2.28,2.46,3.19,3.33,P＜0.05～0.01;突变率:F=2.36,2.61,3.25,3.45,P＜0.05～0.01),高浓度药物耐药菌株的突变株和突变率显著高于低浓度药物耐药菌株,随着不规律用药时间的延长,耐药率和突变率均逐渐上升(耐药率:F=2.77,2.88,P＜0.05;突变率:F=2.72,2.85,P＜0.05).结论 PCR-SSCP是一种快速检测结核杆菌rpoB、katG、rpsL、pncA和embB基因突变敏感、特异的方法.     

荧光PCR熔解曲线法在结核分枝杆菌耐药基因检测中的应用

目的探讨气管镜刷检物实时荧光PCR熔解曲线耐药法检测对复治结核病耐药早期诊断的价值,同时根据支气管刷检深部痰液结核菌耐药检测结果,进一步分析指导可疑耐药结核病治疗,规范临床合理用药,为耐药结核病的诊断及治疗提供科学依据。方法纳入160例结核分枝杆菌(MTB)患者为本研究对象。每例患者采用同一标本进行绝对浓度法和熔解曲线法耐药基因检测,观察熔解曲线法对利福平、异烟肼的检测敏感度、特异度、准确度。结果同一标本应用两种方法检测时,绝对浓度法检出利福平21例耐药,139例敏感;检出异烟肼68例耐药,92例敏感。熔解曲线法检出利福平17例耐药,139例敏感,4例未检出结果;检出异烟肼61例耐药,99例敏感。熔解曲线法对利福平的检测准确度为96.15%、敏感度为87.50%、特异度为98.39%;对异烟腓的检测准确度为95.00%、敏感度为88.57%、特异度为96.80%。两种检验方法对利福平及异烟肼的耐药及敏感率比较,差异均无统计学意义(P>0.05)。结论荧光PCR熔解曲线法能够快速检测出MTB对药物的耐药基因检测,而且有良好的敏感度和特异度,有效节约时间、安全高效,是临床检验及诊治耐药结核的有效手段。     

抗结核药物诱导结核分支杆菌形成L型及其特性的观察分析

目的对抗结核药物诱导结核分支杆菌形成L型及其特性进行观察。方法将结构分支杆菌接种于含有乙胺丁醇、异烟肼、利福平的非高渗透压培养基内,通过显微镜逐日对结核分支杆菌L型的形成情况进行观察。在形成L型后过滤培养物,最终生成结核分支杆菌L型纯培养物同时观察结构分支杆菌特异性基因、抗菌药物敏感性、培养以及形态等方面的情况。结果在常规药物试验浓度下,结核分支杆菌在乙胺丁醇、异烟肼以及利福平作用的过程中均能够形成L型。这些抗结核药物与结构分支杆菌之间的作用不再明显,但对于诺氟沙星、氧氟沙星、红霉素、链霉素等药物依然存在比较大的敏感性。非高渗透压培养基内的结核分支杆菌呈现不规则或圆球形态,成链状排列或单个、成双存在。不运动、不黏附瓶壁沉于培养基底部,革兰染色阳性或阴性,抗酸染色阴性,结构分支杆菌保留了原有的特异性PCR引物基因序列。结论乙胺丁醇、异烟肼与利福平虽然对于结核分支杆菌有着比较良好的杀菌作用,但部分情况下也能够被诱导并形成L型,使以上几种药物失去对于结核疾病的治疗作用。在传代培养环境下,非高渗培养基中的结构分支杆菌L型能够使PCR引物基因序列得到保留,能够通过特异性PCR的方式对分支杆菌进行鉴定。依照各种药物与结核分支杆菌L型之间的敏感性特点,需要在应用抗结核药物的过程中与能够作用于结核分支杆菌L型的抗菌药物联合使用。     

结核分枝杆菌异烟肼耐药基因katG点突变的快速检测

目的：探讨快速检测结核分枝杆菌异烟肼（INH）耐药基因型的分子药敏方法。方法：用聚合酶链反应－单链构象多态性（PCR－SSCP）检测了30株INH敏感菌株及30株耐药株的katG基因，随后用SSCP方法鉴定扩增产物有无突变，以结核分枝杆菌H37Rv作对照。结果：所有INH敏感株均观察到katG基因扩增产物，30株INH耐药株中28株观察到katG基因扩增产物。以H37Rv标准株为对照，30株敏感株SSCP带谱与对照相同；28株INH耐药株中13株与对照相同，15株有不同程度的差异。与药敏试验比较，PCR－SSCP检测INH耐药结核菌的敏感性为57%，特异性为100％。结论：多数结核分枝杆菌耐INH是由于katG基因突变所致，用PCR－SSCP筛选突变株可达到快速检测结核分枝杆菌INH耐药基因型的目的。     

Role of the liver in the disposition of intravenous nitroglycerin in the rat

Previous studies have indicated that the liver is the main site of nitroglycerin (NTG) elimination when the drug is systematically infused. To examine this hypothesis, we measured the apparent systemic clearance (Cls) of nitroglycerin in anesthesized rats receiving a constant intravenous infusion at a dose of 100 micrograms per kg per min. Animals were divided into shunt and sham groups; the former had undergone a portal vein ligation 10 days prior to the study, while the latter was subjected to a sham operation. On the study day, half of the animals of each group also received probenecid at 200 mg/kg, i.v., a drug previously reported to inhibit organic nitrate ester reductase (ONER) activity in rat liver. Arterial NTG samples were obtained at 41, 43 and 45 min of infusion in all four experimental groups; Cls was 439 +/- 32 ml per kg per min (mean +/- S.E.) in sham, 460 +/- 44 in sham and probenecid, 477 +/- 39 in shunt, and 461 +/- 34 in shunt and probenecid animals. During NTG infusion, hepatic blood flow (measured with a constant infusion of indocyanine green) was decreased markedly in shunted rats as was liver/body weight, indicating hepatic atrophy. The specific activity of hepatic ONER was similar in all four groups. In spite of marked differences in hepatic blood flow and hepatic mass, the Cls was similar in all four groups. The liver does not appear to be a major site for the elimination of systemic nitroglycerin as hitherto assumed.     

HACCP在学校集体食堂卫生管理中的应用研究

目的 通过HACCP在学校集体食堂管理中的应用,提高食品卫生水平,保障学生的身体健康。方法 HACCP原则。结果 食品卫生达到较高水平,极大地减少了食物中毒的发生。结论 提示HACCP原则可在学校集体食堂的卫生管理中发挥有效作用。     

半导体激光治疗老年人压疮疗效观察

目的探讨半导体激光局部照射治疗老年人压疮疗效.方法:收集2012年1月-2015年6月48例压疮患者,分为半导体组和常规组各24例.常规组采用常规治疗,半导体组采用半导体激光加常规治疗,10 d为1个疗程,不超过3个疗程.疗程结束后比较两组疗效.结果:半导体组压疮愈显率为83.33％,创面愈合时间为(12.75±5.51)d,常规组分别为54.17％和(19.63±8.65)d,组间差异有统计学意义(P＜0.05),两组均未见不良反应.结论:半导体激光加常规治疗压疮效果肯定,无明显不良反应,且操作简便.     

脑卒中恢复期患者社区综合康复的疗效观察

目的：探讨社区开展康复活动对于脑卒中患者肢体功能康复的影响。方法：选取2014年4至10月参与社区“陈睿健康工作室”康复活动的脑卒中恢复期患者50例作为观察组,选取同期延吉社区卫生服务中心门诊的50例脑卒中恢复期患者作为对照组（进行门诊随访、服药、自我康复）。采用知识、信念、行为（KAP）量表对观察组患者进行问卷调查。采用简化Fugl-Meyer肢体运动功能评分表（Fugl-Meyer assessment of motor function,FAM）比较两组患者入组时和入组6个月时的肢体功能恢复情况及对家庭医生（门诊医师）的满意度。结果：入组6个月时,两组患者的FAM均有所改善,观察组FAM得分[上肢（50.92±7.05）分,下肢（33.01±1.37）分]较对照组[上肢（34.85±6.55）分,下肢（25.20±4.88）分]更高,差异有统计学意义（t上肢=11.8215,t下肢=10.8595,P均＜0.01）。观察组患者的满意率为92%（46/50）,明显高于对照组的30%（15/50,P＜0.05）。结论：在社区开展康复活动可以促进脑卒中患者肢体功能的康复,提高患者的满意度。     

Evaluation of the safety of enalapril in the treatment of heart failure in the very old

Summary We have introduced enalapril, in doses equal to or less than the 2.5 mg currently recommended, as an adjuvant to digoxin and diuretics in 17 patients of mean (SD) age 83 (5) years with severe heart failure. Only eleven patients tolerated its introduction. Unlike those reported in younger patients, all but one of the adverse drug reactions occurred 8 h or more after the first dose. Aged patients started on ACE inhibitors should be observed in hospital until stabilized on a maintenance dose. Three patients had an adverse reaction which differed in nature from those previously reported: acute confusional state, ataxia and mesenteric ischaemia.Ten patients were discharged on 5 mg or 10 mg maintenance doses of enalapril. In nine of them improvement on triple therapy was sustained for a minimum of three months. ACE inhibition was lost in the other patient when her compliance with enalapril therapy fell to around 75%: monitoring compliance is essential when ACE inhibitors are used in low dosages.Enalapril was withdrawn during follow up in three patients because of symptoms of mesenteric ischaemia and in four because of dramatic deterioration of renal function. One of the latter was found subsequently to have severe bilateral atheromatous renal artery stenosis. When isosorbide dinitrate was substituted for enalapril, symptoms of mesenteric ischaemia resolved and renal function returned to baseline. Continuing surveillance for adverse effects is essential in patients of this age group with severe heart failure, and the risk of occult renal artery stenosis requires regular biochemical screening during follow up.The benefit to cost ratio of ACE inhibitors might be improved in aged patients by their use at an earlier stage in the natural history of heart failure, when perfusion of essential organs is not grossly impaired, but carefully monitored trials would be necessary to establish this.     

替普瑞酮治疗老年慢性浅表性胃炎的疗效分析

目的分析替普瑞酮治疗老年慢性浅表性胃炎的疗效。方法100例老年慢性浅表性胃炎患者,通过计算机随机表数字法分为对照组和实验组,每组50例。对照组患者选择西咪替丁作为本次治疗药物,实验组患者选择替普瑞酮作为本次治疗药物。比较两组患者临床疗效、不良反应发生情况以及生活质量。结果实验组治疗总有效率92.00%高于对照组的76.00%,差异具有统计学意义(P<0.05)。实验组不良反应发生率6.00%低于对照组的20.00%,差异具有统计学意义(χ^2=4.332,P=0.037<0.05)。实验组患者治疗后的生活质量评分(68.68±4.11)分高于对照组的(57.73±3.48)分,差异具有统计学意义(t=14.377,P=0.000<0.05)。结论对于老年慢性浅表性胃炎疾病患者,选择替普瑞酮进行治疗不仅疗效更为理想,而且还不易增加患者的用药不良反应,有利于提高患者的生活质量,建议进一步推广。     

Nifedipine in the treatment of hypertension in the elderly

The effect of nifedipine monotherapy, retard tablets, 20 mg bid, was evaluated in 23 hypertensive patients, mean age, 79 +/- 2 years. Twenty-one patients completed an eight-week study. Blood pressure (BP) decreased to 160/90 mm Hg in 15 patients; in four additional patients diastolic BP dropped by 15% to 28%. In a subset of five patients with isolated systolic hypertension, a significant reduction in systolic BP was noted. Side effects were relatively mild and only two patients discontinued the study. The results suggest that nifedipine monotherapy offers an alternative, logic, therapeutic approach to hypertension in the elderly.     

Interactions among the cannabinoids in the antagonism of the abdominal constriction response in the mouse

The ability of ⁹-tetrahydrocannabinol (THC), cannabinol (CBN), cannabidiol (CBD), 11-OH THC and 8,11-diOH THC to antagonise the abdominal constriction response in the mouse induced by formic acid, phenylquinone, 5-hydroxytryptamine, prostaglandin E₁ (PGE₁) and bradykinin was tested. THC was an effective antagonist against all nociceptive agents with an ED₅₀ in all cases between 1.0 and 2.6 mg/kg. CBN, while also effective against all nociceptive agents, was less potent than THC, with an ED₅₀ range between 46.2 and 112.5 mg/kg. CBD in doses as high as 200 mg/kg was without effect. Using PGE₁ as the nociceptive agent, 11-OH THC was equipotent to THC while 8,11-diOH THC was inactive. Naloxone, while able to antagonise the antinociceptive effect of morphine against formic acid-induced writhing, did not reverse the antinociceptive effects of THC. There were no pharmacological interactions between THC, CBD and CBN.     

Variation in the use of biologics in the management of rheumatoid arthritis across the UK

Abstract

Objectives:

Treatment options for rheumatoid arthritis (RA) include conventional synthetic disease-modifying antirheumatic drugs (sDMARDs) and newer biologic DMARDs (biologics). This study describes treatment patterns, adherence to guidance and outcomes at hospital/regional level in the UK.