福氏志贺菌ipaB基因的克隆表达及免疫原性研究

构建福氏志贺菌ipaB基因的原核表达载体,诱导表达并纯化.探讨重组蛋白IpaB经口服免疫小鼠后,机体产生的免疫应答.方法:用PCR扩增ipaB目的基因,克隆至pQE-30表达载体中,转化宿主菌DH5α,经IPTG诱导表达蛋白IpaB;Western blot分析其抗原性.纯化后灌胃免疫小鼠,ELISA法检测血清IgG、IgA、小肠黏液sIgA和粪便sIgA.结果:ipaB基因全长1 743 bp.经SDS-PAGE分析相对分子量(Mr)约为64 000.Western blot分析能与福氏志贺菌全菌抗血清反应.免疫小鼠后,血清IgA、IgG,粪sIgA,肠黏液sIgA明显高于对照组(P＜0.01).结论:成功构建了ipaB基因的原核表达载体并能高效表达,纯化后口服免疫可有效诱导黏膜免疫应答,产生高水平的sIgA,发挥局部黏膜免疫作用,为进一步研究细菌性痢疾的疫苗和发病机制奠定基础.     

肠内免疫营养对烫伤大鼠肠黏膜损伤的保护作用

目的探讨肠内免疫营养（EIN）对烧伤后肠黏膜损伤的保护作用。方法采用30％总体表面积Ⅲ度烫伤大鼠模型。64只Wistar大鼠采用随机数字表法分为EIN组和标准肠内营养（EN）组，另取8只大鼠作为伤前对照。EIN组和EN组给予等热量肠内营养液。分别取伤前对照及EN组、EIN组伤后1、4、7和10d检测肠黏液分泌型免疫球蛋白A（sIgA）浓度及血浆二胺氧化酶（DAO）活性和肿瘤坏死因子-α（TNFa）含量的变化，逆转录一聚合酶链反应（RT—PCR）方法检测肠黏膜组织TNF-αmRNA表达。结果烫伤后EN组和EIN组肠黏液sIgA浓度、血浆DAO活性、TNF-α含量和肠黏膜组织TNFamRNA表达均比伤前显著升高（P〈0.05或P〈0．01）；EIN组肠黏液sIgA浓度、血浆DAO活性和TNF-α含量在伤后4、7和10d，肠黏膜组织TNF-αmRNA的表达于伤后1、4、7和10d均比EN组显著降低（P〈0．05或P〈0．01）。结论与标准EN相比，EIN可明显减轻严重烫伤后肠黏膜损伤，其机制与增加肠黏液sIgA浓度、降低肠黏膜组织TNF-αmRNA的表达有关。     

新生儿脐血T细胞亚群、免疫球蛋白与HBV宫内感染的相关性研究

目的分析T淋巴细胞亚群、免疫球蛋白IgM、IgG、IgA的表达水平与新生儿免疫状态之间的关系。方法依据新生儿脐血中HBV-DNA(Hepatitis B Virus-DNA)及乙肝五项的结果,将研究对象中感染HBV(Hepatitis B Virus)的17例作为宫内感染组,未感染的58例作为宫内未感染组,并选择同期正常孕妇分娩的新生儿43例作为对照组,并采用流式细胞术检测3组新生儿脐血中T细胞亚群及免疫球蛋白IgM、IgG、IgA的表达水平。分析3组脐血T淋巴细胞亚群及免疫球蛋白IgM、IgG、IgA的百分比,并进行统计学分析。结果 HBV宫内感染组新生儿脐血中的CD3+、CD4+阳性率低于宫内未感染组及对照组(P 0.05);CD8+阳性率显著高于HBV宫内未感染组及对照组(P 0.05);HBV宫内感染组新生儿脐血中的IgM阳性率高于宫内未感染组及对照组(P 0.05);IgA、IgG各组间差异无统计学意义(P0.05)。结论宫内感染HBV的新生儿存在T淋巴细胞亚群比例失调,使机体的免疫功能降低,不能很好的清除病原;新生儿宫内感染HBV后,IgM较前升高,提示机体受到抗原刺激后,会产生抗体来进行自我保护;IgA、IgG各组间无明显差异,则提示新生儿免疫功能低下。     

低镁摄入对C57BL/6小鼠肺组织β2受体表达的影响

目的:探讨低镁饮食对小鼠肺组织β2受体(β2AR)表达和β2AR低调节的影响。方法:健康4～6周龄清洁级雌性C57BL/6小鼠96只,体重12±2克,随机分为A、B、C、D组,每组各24只。A、B组予低镁饮食,C、D组予正常镁饮食。B、D组腹腔内注射沙丁胺醇诱导β2受体低调节,A、C组腹腔内注射生理盐水作对照。d1、d21、d34天各组分别随机抽取8只检测血浆Mg2+、红细胞内Mg2+、肺组织β2受体mRNA及蛋白表达。结果:血浆Mg2+、红细胞内Mg2+、肺组织β2ARmRNA及蛋白表达:d1各组间差异无显著性(P>0.05)。d21、d34组间比较:C组显著高于A组(d21:0.95±0.07vs0.73±0.08mmol/L、2.44±0.09vs2.32±0.09mmol/L、0.82±0.10vs0.61±0.04、96.6±7.1vs75.1±4.3fmol/mgprotein;d34:0.94±0.10vs0.69±0.14mmol/L、2.43±0.08vs2.26±0.13mmol/L、0.81±0.08vs0.58±0.06、99.2±7.5vs69.6±7.1fmol/mgprotein)(P均<0.05);D组显著高于B组(d21:0.92±0.09vs0.60±0.10mmol/l、2.46±0.16vs2.13±0.16mmol/L、0.73±0.10vs0.52±0.03、97.4±7.9vs62.3±7.2fmol/mgprotein;d34:0.67±0.10vs0.47±0.08mmol/L、2.07±0.12vs1.89±0.14mmol/L、0.62±0.05vs0.43±0.05、78.3±8.4vs45.1±7.3fmol/mgprotein)(P均<0.05)。结论:细胞内镁不足小鼠肺组织β2AR表达减少;维持一定水平的细胞内镁可抑制激动剂诱导的小鼠肺组织β2AR低调节。     

定期单采血小板捐献者部分免疫指标变化的研究

目的了解定期单采血小板捐献者外周血T淋巴细胞亚群及免疫球蛋白IgG、IgM、IgA水平的变化。方法按捐献间隔期将87名男性单采血小板捐献者分为对照组:首次单采血小板捐献者;实验1组:2次单采血小板间隔45～73 d的捐献者;实验2组:2次单采血小板间隔29～44 d的捐献者。分别用流式细胞术和免疫比浊法检测其外周血T淋巴细胞亚群和免疫球蛋白IgG、IgM、IgA的水平,比较3组捐献者上述指标的差异。结果外周血中CD3+CD4+T淋巴细胞、CD3+CD4+T淋巴细胞/CD3+CD8+T,在实验1组中为(51.33±10.47)%和1.36±0.43,在实验2组中为(50.48±9.50)%和1.34±0.51,在对照组中为(55.18±9.84)%和1.61±0.64。外周血中CD3+CD8+T淋巴细胞、IgG、IgM、IgA,在实验1组中为(39.42±7.14)%、(13.13±2.56)g/L、(1.37±0.60)g/L、(2.55±0.70)g/L,在实验2组中为(40.72±9.24)%、(13.13±2.58)g/L、(1.31±0.70)g/L、(2.44±0.70)g/L,在对照组中为(37.32±8.30)%、(11.55±1.95)g/L、(1.08±0.40)g/L和(2.20±0.91)g/L。3组间上述指标的差异均无统计学意义(P>0.05)。结论定期捐献单采血小板不会影响外周血T淋巴细胞亚群和IgG、IgM、IgA的水平。     

新生儿脐血T细胞亚群、免疫球蛋白与HBV宫内感染的相关性研究

目的分析T淋巴细胞亚群、免疫球蛋白IgM、IgG、IgA的表达水平与新生儿免疫状态之间的关系。方法依据新生儿脐血中HBV-DNA(Hepatitis B Virus-DNA)及乙肝五项的结果,将研究组中乙型肝炎病毒(Hepatitis B Virus,HBV)的17例作为宫内感染组,未感染的58例作为宫内未感染组,并选择同期正常孕妇分娩的新生儿43例作为对照组,并采用流式细胞术检测三组新生儿脐血中T细胞亚群及免疫球蛋白IgM、IgG、IgA的表达水平。分析三组脐血T淋巴细胞亚群及免疫球蛋白IgM、IgG、IgA的百分比,并进行统计学分析。结果 HBV宫内感染组新生儿脐血中的CD3+、CD4+阳性率低于宫内未感染组及对照组(P <0.05);CD8+阳性率显著高于HBV宫内未感染组及对照组(P <0.05);HBV宫内感染组新生儿脐血中的IgM阳性率高于宫内未感染组及对照组(P <0.05);IgA、IgG各组间无明显差距(P>0.05)。结论宫内感染HBV的新生儿存在T淋巴细胞亚群比例失调,使机体的免疫功能降低,不能很好的清除病原;新生儿宫内感染HBV后,IgM较前升高,提示机体受到抗原刺激后,会产生抗体来进行自我保护;IgA、IgG各组间无明显差距,则提示新生儿免疫功能低下。     

重型肝炎患者免疫球蛋白及补体C3的变化

目的了解本地区重型肝炎患者IgA、IgG、IgM及C3变化的临床意义.方法以甲型肝炎为对照组,采用透射免疫比浊法分别对重型肝炎组、对照组进行IgA、IgG、IgM及C3测定.结果重型肝炎组IgA、IgG、IgM及C3分别为(17.78±3.37)g/L、(3.31±0.52)g/L、(2.75±0.62)g/L、(0.65±0.13)g/L,与对照组IgA、IgG、IgM及C3含量(12.56±3.12)g/L、(2.43±0.48)g/L、(1.52±0.64)g/L、(1.05±0.31)g/L相比均有显著性差异(P＜0.05).结论重型肝炎患者多呈高免疫球蛋白血症,且C3明显降低.     

儿童重型β珠蛋白生成障碍性贫血患者外周血淋巴细胞亚群和免疫球蛋白水平状态的研究

目的 研究重型β珠蛋白生成障碍性贫血患儿外周血淋巴细胞各亚群、血清免疫球蛋白(Ig)水平的变化,并探讨其在重型β珠蛋白生成障碍性贫血患儿输血治疗过程中的意义。方法 采用流式细胞术(FCM)和免疫比浊法,对45例重型β珠蛋白生成障碍性贫血患儿和50例健康对照组儿童外周血T淋巴细胞亚群(包括CD3⁺,CD3⁺CD4⁺,CD3⁺CD8⁺和CD4⁺/CD8⁺)、B淋巴细胞亚群(CD3^-CD19⁺)、NK细胞(CD16⁺CD56⁺)和血清Ig(IgG,IgA和IgM)水平进行检测,两组间进行比较。结果 与健康组相比较,重型β珠蛋白生成障碍性贫血患儿CD3⁺T细胞比例[(60.49±9.23)% vs(68.41±6.31)%,t=2.46,P=0.005],CD4⁺T细胞比例[(31.19±5.02)% vs(43.23±4.52)%,t=3.31,P=0.002],CD4⁺/CD8⁺比值[(1.19±0.31)% vs(1.73±0.27)%,t=3.78,P=0.001]、NK细胞[(9.27±2.24)% vs(18.04±5.07)%,t=3.92,P=0.001]比例明显减低,差异均具有统计学意义; 而CD8⁺T细胞比例[(29.11±4.03)% vs(24.49±5.13)%,t=1.89,P=0.04]和CD3^-CD19⁺B细胞比例[(26.13±2.99)% vs(12.53±3.11)%,t=4.45,P=0.001]明显增高,差异均具有统计学意义。与此同时,与健康组相比较,重型β珠蛋白生成障碍性贫血患儿血清IgG(13.27±2.31 g/L vs 11.41±1.81 g/L,t=2.93,P=0.011),IgA(1.99±0.43 g/L vs 1.19±0.19 g/L,t=2.69,P=0.043)和IgM(1.91±0.45 g/L vs 1.49±0.27 g/L,t=2.81,P=0.02)水平明显增高,差异均具有统计学意义。结论 重型β珠蛋白生成障碍性贫血患儿体内存在着细胞和体液免疫功能紊乱。外周血淋巴细胞亚群和免疫球蛋白水平状态的测定,对监测患儿输血治疗过程中机体免疫功能状态的变化有重要意义。     

反复呼吸道感染患儿部分免疫指标的检测

目的观察反复呼吸道感染患儿T淋巴细胞亚群、免疫球蛋白IgG、IgA、IgM及补体C3的变化.结果60例反复呼吸道感染患儿CD3、CD4、CD3/CD8检测结果分别为(58.8±6.8)%、(36.2±3.6)%和1.07±0.14,均明显低于对照组(P＜0.001);CD8为(32±3.8%),明显高于对照组(P＜0.001);IgG、IgA和C3分别为(9.4±1.9)g/L、(0.92±0.18)g/L、(1.21±0.14)g/L均明显低于对照组(P＜0.001);IgM为(0.98±0.17)g/L,与对照组无明显差异(P＞0.05).结论反复呼吸道感染患儿存在细胞和体液免疫功能低下及平衡失调.     

沙参麦冬汤对运动小鼠免疫功能的影响*

目的:探讨沙参麦冬汤对运动小鼠免疫功能的影响。方法:40只雄性昆明种小鼠,随机分为4组：安静对照组、运动对照组、运动+小剂量沙参麦冬汤组、运动+大剂量沙参麦冬汤组,运动小鼠递增负荷游泳训练40d,游泳时间由40min开始渐增至100min,第40天游泳至力竭,24h后处死取血。测定小鼠免疫器官脏器/体重比值、外周血免疫球蛋白、外周血T细胞亚群。结果：服药组小鼠脾指数、外周血IgG、IgA、CD4+T细胞百分比、CD4+/CD8+比值均显著高于运动对照组(P<0.01或P<0.05);大剂量组小鼠胸腺指数、     

Guns and Knives in New Mexico: Patterns of Penetrating Trauma, 1978–1993

Objective: To identify patterns of nonfatal and fatal penetrating trauma among children and adults in New Mexico using ED and medical examiner data.
Methods: The authors retrospectively sampled in 5-year intervals all victims of penetrating trauma who presented to either the state Level-1 trauma center or the state medical examiner from a 16-year period (1978–1993). Rates of nonfatal and fatal firearm and stabbing injury were compared for children and adults.
Results: Rates of nonfatal injury were similar (firearm, 34.3 per 100,000 person-years; stabbing, 35.1). However, rates of fatal injury were significantly different (firearm, 21.9; stabbing, 2.7; relative risk: 8.2; 95% confidence interval: 5.4, 12.5). From 1978 to 1993, nonfatal injury rates increased for children (p = 0.0043) and adults (p < 0.0001), while fatal penetrating injury remained constant. The increase in nonfatal injury in children resulted from increased firearm injury rates. In adults, both stabbing and firearm nonfatal injury rates increased.
Conclusions: Nonfatal injury data suggest that nonfatal violence has increased; fatal injury data suggest that violent death rates have remained constant. Injury patterns vary by age, mechanism of trauma, and data source. These results suggest that ED and medical examiner data differ and that both are needed to guide injury prevention programs.     

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.     

Interprofessional approaches to creating safe,high quality health care

Three supplementary perspectives are presented arguing that interprofessional collaboration is both necessary and desirable. Nonetheless, there are often too many serious intra-professional barriers and obstacles to interprofessional collaboration to make it successful. Some of these barriers, it is argued and illustrated, are found in the multiple ways in which professional identity is tacitly acquired and embodied in the practitioners' habitual, everyday practice. The paper then explores ways in which reflection, especially Second order reflection, can help to elucidate and overcome these obstacles, as well as increasing professional adaptability and competence.     

Systematic Reviews Published in the January 2011 Issue of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD and monthly online. The January 2011 issue (first quarterly DVD for 2011) contains 4515 complete reviews, 1985 protocols for reviews in production, and 13,521 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 641,000 randomized controlled trials, and 14,018 cited papers in the Cochrane methodology register. The health technology assessment database contains over 9300 citations. One hundred and seven new reviews have been published in the last 3 months, of which five have potential relevance for practitioners in pain and palliative medicine.     

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.     

Modified micro-superior percutaneously-assisted total hip: early experiences & case reports

Because of the extensile nature and familiarity of the standard posterior-lateral approach to the hip, a family of "micro-posterior" approaches has been developed. This family includes the Percutaneously-Assisted Total Hip (PATH) approach, the Supercapsular (SuperCap) approach and a newer hybrid approach, the Supercapsular Percutaneously-Assisted Total Hip (SuperPATH) approach. Such approaches should ideally provide a continuum for the surgeon: from a "micro" (external rotator sparing) posterior approach, to a "mini" (external rotator sacrificing) posterior approach, to a standard posterior approach. This could keep a surgeon within his comfort zone during the learning curve of the procedure, while leaving options for complicated reconstructions for the more practiced micro-posterior surgeons. This paper details one author's experiences utilizing this combined approach, as well as permutations of this entire micro-posterior family of approaches as applied to more complex hip reconstructions.     

Structure and function of "metalloantibiotics"

Although most antibiotics do not need metal ions for their biological activities, there are a number of antibiotics that require metal ions to function properly, such as bleomycin (BLM), streptonigrin (SN), and bacitracin. The coordinated metal ions in these antibiotics play an important role in maintaining proper structure and/or function of these antibiotics. Removal of the metal ions from these antibiotics can cause changes in structure and/or function of these antibiotics. Similar to the case of "metalloproteins," these antibiotics are dubbed "metalloantibiotics" which are the title subjects of this review. Metalloantibiotics can interact with several different kinds of biomolecules, including DNA, RNA, proteins, receptors, and lipids, rendering their unique and specific bioactivities. In addition to the microbial-originated metalloantibiotics, many metalloantibiotic derivatives and metal complexes of synthetic ligands also show antibacterial, antiviral, and anti-neoplastic activities which are also briefly discussed to provide a broad sense of the term "metalloantibiotics."