结核分枝杆菌培养和药敏试验及菌群鉴定快速荧光法的研究

目的 建立一种经济、快速的结核分枝杆菌培养、药敏和菌群鉴定的方法。方法 用我们研制的快速荧光检测法（本法）与传统的改良罗氏法（L-J法），对分枝杆菌标准菌株及临床分离菌株，同期进行培养、药敏试验和菌群鉴定的对照研究，验证本法培养结核分枝杆菌的特性，确定药敏和菌群鉴定的药物最适应用浓度。结果 本法培养比L-J法检出时间平均提前19．13d，且阳性率高。药敏试验和菌群鉴定中各药物浓度确定为INH0．1μg／ml、RFP1μg／ml、EB2μg／ml、AMK2μg／ml、LVFX2μg／ml、PNB200μg／ml和TCH2、5μg／ml；7～10d可获结果，比L-J法缩短18～21d。结论 本法明显缩短了结核分枝杆菌培养、药敏和菌群鉴定的时间，提高培养阳性率；经济、实用，适合在基层单位推广。     

结核分枝杆菌临床分离株药敏结果与耐药程度的关联分析

目的 探讨结核分枝杆菌临床分离株对12种抗结核药物的MIC值及其药敏检测结果 与耐药程度的关联规律,从而为临床制定治疗方案提供借鉴依据.方法 采用液体培养基联合MTT技术进行抗结核药物的MIC检测.对上海市肺科医院2009年1-6月间的163株结核分枝杆菌临床分离株进行RFP、INH、SM、EMB、OFLX、LVFX、MOX、AMK、CPM、PTA、CLA和PAIN的MIC检测和Bactec MGIT快速培养仪药敏检测,并进行MIC与耐药程度的关联性分析.结果 67%(42/62)的SM耐药株MIC≥16 μg/ml,63%(51/81)的INH耐药株MIC≥8 μg/ml,77%(50/65)的RFP耐药株MIC≥8 μg/ml,20%(12/60)的EMB耐药株MIC≥4 μg/ml;43%(25/58)的OFLX耐药株MIC≥8 μg/ml;41%(15/37)的AMK耐药菌株MIC≥16 μg/ml,41%(12/29)的CPM耐药菌株MIC≥4 μg/ml.OFLX耐药株的3个氟喹诺酮类药物的MIC(OFLX、LVFX和MOX的MIC分别为2～128、1～32和0.0625～1 μg/ml)差异有统计学意义(F=16.874,P<0.01);SM、INH、RFP、EMB、OFLX、AMK和CPM在任6种及7种药物同时耐药株中的MIC值(分别为0.5～128、2～64、0.25～128、1～32、1～64、0.5～128和1～128μg/ml)明显高于任1种及2种药物耐药菌株的MIC值(MIC值分别为0.25～128、0.0625～64、0.25～32、0.25～2、0.125～2、0.5～4和1～4 μg/ml,F值分别为20.066、40.499、47.197、70.373、91.432、41.840和21.547,P均<0.05);SM、INH、RFP、EMB在4种药物同时耐药菌株中MIC值(分别为1～128、2～64、0.25～128和1～32μg/ml)显著高于任1种及2种药物耐药株(MIC值分别为0.25～128、0.0625～64、0.25～64和0.25～2 μg/ml,F值分别为26.242、23.563、31.541和64.469,P均<0.05);RFP在MDR酶株的MIC值(2～64 μg/ml)显著高于非MDR的耐药菌株的MIC值(0.25 μg/ml,F=5.613,P<0.05).结论 本研究揭示了常用的12种抗结核药物的耐药程度与常规药敏检测结果 之间的关联规律,为临床医生根据常规药敏结果 制定更加有效的抗结核治疗方案提供重要的借鉴.     

甘肃省53株非结核分枝杆菌对9种常用抗结核药物的敏感性分析

目的 初步了解2012－2014年甘肃省临床分离的非结核分枝杆菌（NTM）对常用抗结核药物的耐药情况。方法 收集临床分枝杆菌分离株,用PNB/TCH生长试验和基于16S rRNA基因的序列分析法鉴定NTM。采用传统固体罗氏培养基绝对浓度法对NTM进行9种常用抗结核药物链霉素（SM）、利福平（RFP）、丙硫异烟胺（TH₁₃₂₁）、异烟肼（INH）、对氨基水杨酸钠（PAS）、左氧氟沙星（LVFX）、乙胺丁醇（EMB）、卡那霉素（KAN）和吡嗪酰胺（PZA）的敏感性测试。结果 共收集到分枝杆菌1 081株,经鉴定53株为NTM,分离率为4.90%。其中胞内分枝杆菌最多,为36株;其次是堪萨斯分枝杆菌,为8株;其他依次是鸟分枝杆菌4株、戈登分枝杆菌2株、脓肿分枝杆菌2株和偶然分枝杆菌1株。药敏结果显示,耐药率从高到低依次为RFP（73.58%）、TH₁₃₂₁（69.81%）、INH（67.92%）、SM（62.26%）、PAS（60.38%）、PZA（47.17%）、EMB（28.30%）、LVFX（26.41%）、KAN（15.09%）,其中耐药株49株,耐药率为92.45%,NTM对9种抗结核药物的耐药程度有差异。结论 NTM对抗结核药物敏感性差,应参照NTM药敏结果选择合理的药物,提高治愈率,减少耐药发生。     

抗酸分枝杆菌菌种鉴定的快速、微量、直视方法

目的创建一种经济、适用、快速、微量、直视的抗酸分枝杆菌菌种鉴定系统(以下简称"本法")。方法利用对硝基苯甲酸(PNB)对结核分枝杆菌有抑菌作用而对非结核分枝杆菌无抑菌作用的原理,对分枝杆菌标准菌株及临床分离菌株进行PNB抑菌试验,以改良罗氏法(L-J法)对照,确定药敏试验最适的PNB应用浓度。利用自行研制的电脑控制无级连续超倍放大、微量培养和图象采集系统,建立分枝杆菌菌种快速、微量、直视鉴定系统。结果本法PNB的应用浓度为400μg/L;检出时间平均5.6d。每实验孔仅需培养基100μl。本法结果与L-J法结果符合率达94.6%。结论本法明显缩短了分枝杆菌鉴定的报告时间,节省试剂,简单、经济、实用。     

结核分枝杆菌快速微量直视培养药敏试验方法的研究

目的建立一种经济、适用、快速的微量结核分枝杆菌（MTB）直视培养药敏方法。方法研制电脑控制无级连续超倍放大、微量培养和图像采集组成的MTB快速培养药敏及鉴定系统;用该系统进行MTB标准菌株H37Rv及临床分离菌株药敏试验;以改良罗氏法（L—J法）为对照,确定药敏试验药物最适浓度。结果本法药敏试验的药物：硫酸链霉素、异烟肼、利福平、乙胺丁醇、硫酸阿米卡星、硫酸卷曲霉素、盐酸左氧氟沙星及对氨基水杨酸异烟肼应用浓度分别为：1、0．1、1、5、5、10、4、1μg／ml;检出时间H37Rv为3d;临床分离菌165株平均5．6d。每实验孔仅需培养基100μl。结论快速法显著缩短了MTB药敏试验时间,节省试剂,操作简便,适合基层单位应用。     

50例分枝杆菌药敏实验的意义

[目的]对结核分枝杆菌及非典型分枝杆菌培养阳性药物的敏感性及耐药性研究及意义。[方法]用生物梅里埃BacT／ALERT药敏试剂（内含链霉素SM、异烟肼INH、利福平RFP、乙胺丁醇EMB4种药）对50例分枝杆菌培养阳性的标本做药敏试验。[结果]通过药敏试验，得出痰、尿、椎体脓液等临床标本对SM、INH、RFP、EMB的敏感性及耐药性比例，以及结核分枝杆菌（TB）及非典型分枝杆菌（MOIT）的耐药程度。[结论]通过分枝杆菌药敏试验对指导帮助临床用药有一定的意义。     

噻唑蓝和刃天青法在一线抗结核药物药敏试验中的应用

目的 评价噻唑蓝[3-(4,5-二甲基噻唑)-2,5-二苯基溴化四氮唑蓝,MTT]和刃天青(resazurin)2种快速而价廉的药敏试验方法进行一线抗结核药物的药敏试验的可行性.方法 分别用MTT和刃天青2种微孔板方法对64株结核临床分离菌株进行一线抗结核药物的药敏检测,并将结果与传统罗氏培养基的绝对浓度法的结果进行比较分析.结果 MTT和刃天青2种微孔板方法所得利福平(RFP)、异烟肼(INH)、乙胺丁醇(EMB)、链霉素(SM)4种一线抗结核药物的药敏结果与传统绝对浓度法有较好的一致性,MTT法进行RFP、INH、EMB、SM药敏试验的敏感度分别为94.8%、93.8%、92.9%、90.6%,特异度分别为96.0%、93.8%、96.0%、87.5%,准确性分别为95.3%、93.8%、95.3%、89.1%;刃天青法进行BFP、INH、EMB、SM药敏试验的敏感度分别为92.3%、90.6%、92.9%、87.5%,特异度分别为96.0%、90.6%、94.0%、87.5%,准确性分别为93.8%、90.6%、93.8%、87.5%.4种一线抗结核药物应用MTT法与绝对浓度法的Kappa值为:RFP 0.857,INH 0.831,EMB 0.714,SM 0.792;应用刃天青法与绝对浓度法的Kappa值为:RFP 0.871,INH 0.826,EMB 0.826,SM 0.750.MTT和刃天青2种药敏方法4种一线抗结核药物的Kappa值为:RFP 0.889,INH 0.875,EMB 0.787,SM 0.844.结论 MTT和刃天青是2种简单快速、价廉且敏感的药敏方法,有希望成为一线抗结核药物药敏试验的候选方法.     

刃天青显色法检测结核分枝杆菌的耐药性

目的用刃天青显色法检测结核分枝杆菌对4种一线抗结核药物的耐药性,并探讨该法的临床应用价值。方法来自河南省3家结核病医院的共240株结核分枝杆菌分别用刃天青显色法检测异烟肼(INH)、利福平(RFP)、乙胺丁醇(EMB)、硫酸链霉素(SM)的耐药性,并与比例法作比较。结果刃天青显色法检测平均时间为8d,与比例法相比结果一致性好,INH、RFP、EMB、SM 4种药物的折点浓度分别为0.25、2.0、4.0、2.0μg/mL。结论刃天青显色法检测结核分枝杆菌的耐药性具有成本低、快捷、操作简便、准确等优点,为结核耐药检测提供了一种较好的方法。     

西藏地区169例复治失败肺结核病例耐药状况检测

目的 了解西藏地区复治肺结核患者中的结核分枝杆菌对抗结核化学药物的耐药状况,为有效防治肺结核病提供科学依据.方法 收集西藏自治区结核病防治所2000-2005年复治失败病例169例痰标本,用改良罗氏培养基分离培养结核分枝杆菌,绝对浓度法检测分离菌株对4种抗结核药物[链霉素(SM)、异烟肼(INH)、乙氨丁醇(EMB)、利福平(RFP)]的耐药性测定.结果 共检测169株结核分枝杆菌,其中耐药株165株,总耐药率高达97.63%,其中耐多药(所耐药物中含R和H)133例,耐多药率为78.70%.复治失败的耐药率和耐多药率明显高于全国46.50%的水平.对RFP、INH、SM和EMB的耐药率分别为91.12%(154/169)、84.02%(142/169)、75.15%(127/169)和21.30%(36/169).结论 西藏地区属高耐药区,应加强结核分枝杆菌的耐药监测和控制.     

南京地区结核分枝杆菌耐药状况的调查

目的监测结核分枝杆菌对一线和二线抗结核药物的耐药水平,掌握耐多药结核病流行情况。方法2003~2008年6年内本院收集的4 703株临床分枝杆菌,采用绝对浓度法在改良罗氏培养基上检测其对4种一线抗结核药和5种二线抗结核药的敏感性。结果结核复合群4 470株(95%),非结核分枝杆菌233株(5%)。4 703株分枝杆菌对9种抗结核药的耐药率为RIF(48.50%)、INH(45.52%)、SM(31.85%)、EB(19.58%)、DIP(18.56%)、LVX(16.24%)、PAS(9.78%)、AMK(7.61%)、CPM(6.29%)。与2003年的报道相比,对INH、RIF、EB、LVX、DIP的耐药率均明显增高,P<0.01;对SM、CPM的耐药率增高无显著性差异,P>0.05;而对AMK和PAS的耐药率明显降低,P<0.01。4 470株结核杆菌中有耐多药结核菌(MDR-TB)和严重耐多药结核菌(XDR-TB)分别为1 573和136株,分别占结核杆菌总数的35.19%和3.04%。其中MDR-TB数比2003年报道的32.02%略高。结论结核杆菌对抗结核药耐药水平较高且有增高的趋势,MDR-T...     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

Dissociable correlates of two classes of retrieval processing in prefrontal cortex

Although substantial evidence suggests that the prefrontal cortex (PFC) implements processes that are critical for accurate episodic memory judgments, the specific roles of different PFC subregions remain unclear. Here, we used event-related functional magnetic resonance imaging to distinguish between prefrontal activity related to operations that (1) influence processing of retrieval cues based on current task demands, or (2) are involved in monitoring the outputs of retrieval. Fourteen participants studied auditory words spoken by a male or female speaker and completed memory tests in which the stimuli were unstudied foil words and studied words spoken by either the same speaker at study, or the alternate speaker. On "general" test trials, participants were to determine whether each word was studied, regardless of the voice of the speaker, whereas on "specific" test trials, participants were to additionally distinguish between studied words that were spoken in the same voice or a different voice at study. Thus, on specific test trials, participants were explicitly required to attend to voice information in order to evaluate each test item. Anterior (right BA 10), dorsolateral prefrontal (right BA 46), and inferior frontal (bilateral BA 47/12) regions were more active during specific than during general trials. Activation in anterior and dorsolateral PFC was enhanced during specific test trials even in response to unstudied items, suggesting that activation in these regions was related to the differential processing of retrieval cues in the two tasks. In contrast, differences between specific and general test trials in inferior frontal regions (bilateral BA 47/12) were seen only for studied items, suggesting a role for these regions in post-retrieval monitoring processes. Results from this study are consistent with the idea that different PFC subregions implement distinct, but complementary processes that collectively support accurate episodic memory judgments.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

Clinical Pharmacokinetics of Morphine

Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.