我国结核病免疫学预防、治疗与诊断用制品发展进程与展望

结核病的免疫学预防、治疗与诊断是控制结核病的最重要措施之一。作者总结了我国结核病免疫学生物制品的发展进程、研究现状及未来的发展方向,同时展望了在已构建的结核病免疫预防体系框架下,有机运用各种生物制品,降低我国结核感染率、发病率,改变我国结核病高负担现状的可能性。     

Liver transplantation: making the best out of what we have

Since the first human liver transplantation, carried out in 1963, the procedure has become routine with an excellent outcome in terms of both quality and length of survival. One of the major challenges facing the transplant community is the shortage of donor organs. Possible approaches to overcoming this problem include changes in legislation, setting up of organizational structures, more effective use of marginal donor livers, splitting livers, and the development of living related transplants. Alternative treatments to liver transplantation have been sought, including hepatocyte transplantation, xenotransplantation, liver-directed gene therapy, extracorporeal liver support systems, tissue-engineered organs, and auxiliary partial orthotopic liver transplantation.     

骨髓基质细胞不同培养方法效果比较

目的：观察全骨髓培养法与密度梯度离心法对培养骨髓基质细胞的效果差异,建立简单、经济、实用分离骨髓间充质干细胞方法。方法：体外提取骨髓基质细胞,分别采用全骨髓培养法与密度梯度离心法培养骨髓基质细胞,观察细胞形态学、细胞活力、细胞周期及细胞表面分子表达差异。结果：两种方法均能有效培养出骨髓间充质干细胞,各细胞周期的细胞检出率无显著差异（P〉0．05）,细胞形态学无差异,细胞表面分子CD34、SH1、SH2、SH3表达差异不显著（P〉0．05）。结论：全骨髓培养法在分离培养骨髓基质细胞方面与密度梯度离心法具有类似效果。     

鄱阳湖区洲滩植被与钉螺空间分布关系的研究

目的探讨鄱阳湖区洲滩植被分布与钉螺孳生地的空间分布规律,以更好地利用遥感技术对洲滩钉螺孳生地进行监测。方法选择鄱阳湖区血吸虫病流行村附近的洲滩为调查点,用传统方法进行植被和钉螺调查,卫星定位仪（GPS）记录调查点的经纬度,并利用ArcGIS软件把地面调查数据矢量化。运用遥感图像处理软件PCI9.0,对鄱阳湖区春季枯水期的卫星TM图像进行几何校正和非监督分类。然后,把矢量化的地面数据与卫星图片叠加,进行钉螺孳生地与植被关系的空间分布分析。结果鄱阳湖区洲滩植被可以被划分成3类地表类别,其中单一优势植被区为钉螺主要孳生类别,湿生植被区和混合杂草区为少螺或无螺类别。结论洲滩植被分布规律与钉螺孳生地关系密切,呈片状和带状分布,应用遥感技术能快速确定鄱阳湖区洲滩的钉螺孳生地。     

HIV实验室检测技术进展

HIV实验室检测技术对AIDS的诊断、治疗及疫情监测极其重要。随着AIDS防治工作的不断深化,实验室检测技术也在不断发展,不仅抗原抗体检测、核酸检测及免疫学检测等常规技术得到了进一步完善,还出现了BED捕获酶免疫试验、滤纸干血片技术、新生儿早期诊断及超敏反应基因检测等以更好监测、更早诊断及更好治疗为目的的新技术。本文对HIV实验室检测技术进展进行了阐述。     

Expert consensus for the diagnosis and treatment of cystic and alveolar echinococcosis in humans

The earlier recommendations of the WHO-Informal Working Group on Echinococcosis (WHO-IWGE) for the treatment of human echinococcosis have had considerable impact in different settings worldwide, but the last major revision was published more than 10 years ago. Advances in classification and treatment of echinococcosis prompted experts from different continents to review the current literature, discuss recent achievements and provide a consensus on diagnosis, treatment and follow-up. Among the recognized species, two are of medical importance - Echinococcus granulosus and Echinococcus multilocularis - causing cystic echinococcosis (CE) and alveolar echinococcosis (AE), respectively.For CE, consensus has been obtained on an image-based, stage-specific approach, which is helpful for choosing one of the following options: (1) percutaneous treatment, (2) surgery, (3) anti-infective drug treatment or (4) watch and wait. Clinical decision-making depends also on setting-specific aspects. The usage of an imaging-based classification system is highly recommended.For AE, early diagnosis and radical (tumour-like) surgery followed by anti-infective prophylaxis with albendazole remains one of the key elements. However, most patients with AE are diagnosed at a later stage, when radical surgery (distance of larval to liver tissue of >2 cm) cannot be achieved. The backbone of AE treatment remains the continuous medical treatment with albendazole, and if necessary, individualized interventional measures. With this approach, the prognosis can be improved for the majority of patients with AE.The consensus of experts under the aegis of the WHO-IWGE will help promote studies that provide missing evidence to be included in the next update.     

Experimental canine leishmaniasis: Clinical, parasitological and serological follow-up

Canine leishmaniasis (CanL) caused by Leishmania infantum is transmitted by the bite of phlebotomine sand flies and affects millions of dogs in Europe, Asia, North Africa and South America. Canis familiaris is the major host for these parasites, and the main reservoir for human visceral infection. The development of effective molecules for therapy and immunoprophylaxis, would be an important tool in the control of this zoonosis. The aim of this study was to characterize an experimental CanL model in order to determine the best challenge model and which parameters are the most reliable to evaluate the efficacy of new drugs or vaccine candidates against L. infantum infection. The intravenous challenge with purified amastigotes used in this study allowed the development of infection in all animals inoculated (as confirmed by the detection of parasite in the different tissues and organs collected 6 months after inoculation). Molecular and serologic techniques were efficient methods for the follow-up. Lymph node and bone marrow aspirates were suitable clinical samples to detect the presence of Leishmania parasites. Despite ELISA was highly sensitive in detecting specific anti-Leishmania antibodies the use of two tests can improve the sensitivity and specificity of serological diagnosis.     

人外周血树突状细胞体外稳定培养方法的建立及与磁珠法的比较

目的 建立一种稳定的人外周血树突状细胞(DCs)体外培养的方法,并与磁珠分选法进行比较.方法 通过密度梯度离心法分离出志愿者的外周血单个核细胞(PBMC),再分别应用磁珠分选法、贴壁法对PBMC进行培养,应用重组人集落刺激因子(rhGM-CSF)、重组人白细胞介素-4(rhIL-4)诱导获得DCs.倒置显微镜观察细胞形态变化,并分别在第3、5、6天用台盼蓝染色法进行细胞活力检测;经过1、2、5 h的贴壁培养后,应用流式细胞仪检测单核细胞表面CD14、CD1a、HLA-DR的表达以确定最佳贴壁时间;经人重组细胞因子诱导培养后,对所获得的细胞检测CD14、CD1a、CD86、CD83、HLA-DR的表达.采用同种混合淋巴细胞反应,评价DCs刺激T淋巴细胞增殖的能力.结果 经贴壁2 h后诱导培养的DCs形态较典型.磁珠分选法获得的DCs第5、6天细胞活力[(53.333±5.774)%、(38.333±7.638)%]明显低于第3天[(68.667±3.215)%,P均＜0.05];贴壁培养法获得的DCs第3、5、6天的细胞活力[(92.667±3.055)%、(94.000±1.000)%和(94.667±1.528)%]比较,差异无统计学意义(F=0.737,P＞0.05);贴壁培养法获得的DCs第3、5、6天细胞活力均高于磁珠分选法(t值分别为9.374、12.021、12.527,P均＜0.05).PBMC经磁珠分选前后CD14的阳性表达率分别为(32.457±12.351)%、(41.914±14.858)%,二者比较差异无统计学意义(t=1.295,P＞0.05).单核细胞表面CD14的阳性表达率在培养2 h时[(35.267±4.658)%]高于培养1、5 h时[(15.033±6.189)%、(21.233±4.895)%,P均＜0.05].培养第6天,DCs表面CD14的阳性表达率[(2.200±1.356)%]较第1天[(32.328±14.517)%]明显下降(t=5.467,P＜0.05),CD1a的阳性表达率[(43.371±16.250)%]较第1天[(12.300±6.223)%]显著升高(t=2.545,P＜0.05);而CD86、CD83、HLA-DR的阳性表达率[(16.857±5.686)%、(9.343±5.230)%、(72.800±17.881)%]与第1天[(12.550±16.758)%、(6.250±1.323)%、(64.671±15.588)%]比较,差异无统计学意义(t值分别为0.652、1.137、0.907,P均＞0.05).同种混合淋巴细胞反应,随着淋巴细胞的增多,增殖能力下降.磁珠分选法中,DCs与淋巴细胞的比例为1:50、1:100时,细胞增殖能力(1.502±0.055、1.507±0.029)较1:10时(1.859±0.049)降低(P均＜0.05);贴壁培养法中,DCs与淋巴细胞的比例为1:100时,细胞增殖能力(1.545±0.066)较1:10时(2.015±0.301)降低(P＜0.05).在DCs与淋巴细胞的比例相同时,两种方法得到的DCs在刺激T淋巴细胞方面的能力相近(P＞0.05).结论 与磁珠分选法比较,贴壁培养2 h后的人外周血PBMC再行诱导可获得形态与功能较优的DCs,且此法稳定、简便、经济,是一种适于基础、临床研究的DCs体外培养方法.     

Electromagnetic navigation transthoracic needle aspiration for the diagnosis of pulmonary nodules: a safety and feasibility pilot study

Background

Pulmonary nodules remain a diagnostic challenge for physicians. Minimally invasive biopsy methods include bronchoscopy and CT guided transthoracic needle aspiration (TTNA). A novel electromagnetic guidance transthoracic needle aspiration (ETTNA) procedure which can be combined with navigational bronchoscopy (NB) and endobronchial ultrasound (EBUS) in a single setting has become available.

Methods

A prospective pilot study examining the safety, feasibility and diagnostic yield of ETTNA in a single procedural setting. All patients enrolled underwent EBUS for lung cancer staging followed by NB and ETTNA. Feasibility of performing ETTNA and a safety assessment by recording procedural related complications including pneumothorax or bleeding was performed. Diagnostic yield of ETTNA defined by a definitive pathologic tissue diagnosis was recorded. An additional diagnostic yield analysis was performed using a cohort analysis of combined interventions (EBUS + NB + ETTNA). All non-diagnostic biopsies were either followed with radiographic imaging or a surgical biopsy was performed.

Results

Twenty-four subjects were enrolled. ETTNA was feasible in 96% of cases. No bleeding events occurred. There were five pneumothoraces (21%) of which only two (8%) subjects required drainage. The diagnostic yield for ETTNA alone was 83% and increased to 87% (P=0.0016) when ETTNA was combined with NB. When ETTNA and NB were performed with EBUS for complete staging, the diagnostic yield increased further to 92% (P=0.0001).

Conclusions

This is the first human pilot study demonstrating an acceptable safety and feasibility profile with a novel ETTNA system. Further studies are needed to investigate the increased diagnostic yield from this pilot study.     

替米沙坦对电压依赖性的Kv1.3和Kv1.5通道电流阻断作用的实验研究

目的 通过观察替米沙坦对电压依赖性的Kv1.3和Kv1.5的阻断作用,探讨替米沙坦对此类通道的阻断可能具有的临床作用.方法 使用双电极电压钳技术记录表达于非洲爪蟾卵母细胞的Kv1.3和Kv1.5钾通道电流,不同浓度灌流观察其对电流影响.结果 (1)替米沙坦浓度依赖性的阻断Kv1.3通道,其阻断的IC50是2.05 μmol/L.替米沙坦对Kv1.3电流的阻断具有电压依赖性.(2)替米沙坦浓度依赖件的阻断Kv1.5通道,其阻断的IC50是2.37 μmol/L.替米沙坦对Kv1.5电流的阻断具有更显著的电压依赖性.结论 替米沙坦阻断开放状态的Kv1.3可能是其发挥免疫调节和抗动脉粥样硬化作用的机制之一.替米沙坦对开放状态的Kv1.5钾通道的阻断可能是其减少心房颤动发生率的作用机制之一.