五种布鲁氏菌核酸检测试剂盒检测性能的评价北大核心CSCD

目的比较5种布鲁氏菌核酸实时荧光PCR检测试剂盒的一致性和检出能力,为临床实验室选择检测方法和布鲁氏菌的诊断提供参考依据。方法选用经病原学检测确定为布鲁氏菌阳性的血液样本38份,健康人的血液样本24份,潘氏变形杆菌、溶藻弧菌、河弧菌、铜绿假单胞菌、肺炎克雷伯菌DNA各1份,使用5种试剂盒(编号A-E)分别进行核酸检测,比较5种试剂盒临床样本检测的一致性;选择1份阳性样本核酸用无RNA酶水梯度稀释得到5个浓度(浓度1:4453.13 fg/μL,浓度2:1113.28 fg/μL,浓度3:278.32 fg/μL,浓度4:69.58 fg/μL,浓度5:17.40 fg/μL),每个浓度使用5种试剂盒(编号A-E)分别进行3次检测,比较5种试剂盒的阳性检出率及批内重复性。结果5种试剂盒检测67份DNA样品的符合率稍有不同,试剂盒ABDE的符合率均为100%,试剂盒C的符合率为98.51%。批内重复性显示5种试剂盒在浓度1、浓度2、浓度3水平重复检测DNA的Ct值变异系数均<5%;在浓度1与浓度4梯度区间,试剂盒的阳性检出能力比较显示试剂盒A、B、D较高,为11/12,试剂盒C和E较低,为8/12。结论5种试剂盒的真实性和可靠性较好,灵敏度和符合率稍有差别,特异度均为100%;重复性较好,检测性能良好。部分试剂盒对弱阳性样本的检出能力不强,该类样本可使用多种试剂盒复核,以保障结果的准确性。     

Epstein-Barr virus (EBV)-associated lymphoproliferations in severe combined immunodeficient mice transplanted with Hodgkin's disease lymph nodes: implications of EBV-positive bystander B lymphocytes rather than EBV-infected Reed-Sternberg cells

To establish an in vivo model for the study of Hodgkin's disease and Reed-Sternberg (RS) cells, 25 lymph node tissue samples involved by Hodgkin's disease were grafted into severe combined immunodeficiency (SCID) mice. Ten Epstein-Barr virus (EBV)-associated tumors were obtained in SCID mice. EBV-positive tumors growing in SCID mice were correlated with the presence of EBV-positive nonneoplastic B cells in patient tumors (90% v 26.6%; P<.01) and was independent of the EBV status of RS cells. Our results suggested that EBV-positive tumors growing in SCID mice originated from normal EBV-positive small lymphocytes (bystander B lymphocytes). We also compared the characteristics of these tumors with those obtained after transplantation of 15 non-Hodgkin's lymphoma and four reactive lymph nodes. The latent period to observe a growing tumor in SCID mice was similar between the two groups (12.86 +/- 5.59 weeks for Hodgkin's disease v 13.6 +/- 5.36 weeks for non-Hodgkin's lymphoma and reactive lymph nodes). The relatively high number of EBV-positive small lymphocytes detected in Hodgkin's disease and T-cell lymphoma compared with B-cell lymphoma may account for the greater percentage of EBV- positive tumors obtained in SCID mice. Our results show that SCID mice do not provide the growth conditions that are required for in vivo growth of RS cells. We noted in some SCID tumors, the presence of binucleated and/or multinucleated giant cells resembling RS cells. However, the presence of such cells was not restricted to mice grafted with lymph nodes involved by Hodgkin's disease. We postulate that in previous reports, cells resembling RS cells were just binucleated EBV- positive lymphoma blastoid cells rather than actual RS cells.     

Comparison of ultrashort-lived iridium-191m with technetium-99m for first pass radionuclide angiocardiographic evaluation of right and left ventricular function in adults

Ultrashort-lived iridium-191m (Ir-191m, physical half-life = 5.0 seconds) has been used in angiocardiography, primarily in pediatric patients. A theoretical obstacle to more widespread use of Ir-191m is the belief that its physical half-life is too short to permit evaluation of left ventricular function in adult patients. To evaluate its usefulness in adults, first pass ejection fractions of the left and right ventricles determined with use of Ir-191m and technetium-99m (Tc-99m) were compared in 33 adult patients. An osmium-191m----iridium-191m (Os-191----Ir-191m) generator was employed to deliver doses of 150 to 250 mCi (5.5 to 9.2 GBq) of Ir-191m for intravenous injection. The whole body radiation absorbed dose with Ir-191m was 15 to 25 mrad. High quality angiocardiograms were obtained with both Tc-99m and Ir-191m. Total counts per image for the right ventricle were 51,000 +/- 8,000 (mean +/- SD) for Ir-191m and 30,000 +/- 8,000 for Tc-99m. The left ventricular counts were comparable for both radiotracers (25,000 +/- 7,000 for Ir-191m and 25,000 +/- 8,000 for Tc-99m). Right ventricular ejection fractions were similar: 44 +/- 8% for Ir-191m and 47 +/- 9% for Tc-99m. The correlation coefficient was 0.93 with a standard deviation of the regression of 3.1% ejection fraction units. The left ventricular ejection fractions were also similar: 45 +/- 14% for Ir-191m and 46 +/- 13% for Tc-99m. The left ventricular ejection fraction correlation coefficient was 0.96 with a standard deviation of the regression of 3.7%.(ABSTRACT TRUNCATED AT 250 WORDS)     

RyR1 Deficiency in Congenital Myopathies Disrupts Excitation–Contraction Coupling

In skeletal muscle, excitation–contraction (EC) coupling is the process whereby the voltage‐gated dihydropyridine receptor (DHPR) located on the transverse tubules activates calcium release from the sarcoplasmic reticulum by activating ryanodine receptor (RyR1) Ca²⁺ channels located on the terminal cisternae. This subcellular membrane specialization is necessary for proper intracellular signaling and any alterations in its architecture may lead to neuromuscular disorders. In this study, we present evidence that patients with recessive RYR1‐related congenital myopathies due to primary RyR1 deficiency also exhibit downregulation of the alfa 1 subunit of the DHPR and show disruption of the spatial organization of the EC coupling machinery. We created a cellular RyR1 knockdown model using immortalized human myoblasts transfected with RyR1 siRNA and confirm that knocking down RyR1 concomitantly downregulates not only the DHPR but also the expression of other proteins involved in EC coupling. Unexpectedly, this was paralleled by the upregulation of inositol‐1,4,5‐triphosphate receptors; functionally however, upregulation of the latter Ca²⁺ channels did not compensate for the lack of RyR1‐mediated Ca²⁺ release. These results indicate that in some patients, RyR1 deficiency concomitantly alters the expression pattern of several proteins involved in calcium homeostasis and that this may influence the manifestation of these diseases.     

Diagnosis and management of asthma in preschoolers: A Canadian Thoracic Society and Canadian Paediatric Society position paper

Asthma often starts before six years of age. However, there remains uncertainty as to when and how a preschool-age child with symptoms suggestive of asthma can be diagnosed with this condition. This delays treatment and contributes to both short- and long-term morbidity. Members of the Canadian Thoracic Society Asthma Clinical Assembly partnered with the Canadian Paediatric Society to develop a joint working group with the mandate to develop a position paper on the diagnosis and management of asthma in preschoolers.In the absence of lung function tests, the diagnosis of asthma should be considered in children one to five years of age with frequent (≥8 days/month) asthma-like symptoms or recurrent (≥2) exacerbations (episodes with asthma-like signs). The diagnosis requires the objective document of signs or convincing parent-reported symptoms of airflow obstruction (improvement in these signs or symptoms with asthma therapy), and no clinical suspicion of an alternative diagnosis. The characteristic feature of airflow obstruction is wheezing, commonly accompanied by difficulty breathing and cough. Reversibility with asthma medications is defined as direct observation of improvement with short-acting ß₂-agonists (SABA) (with or without oral corticosteroids) by a trained health care practitioner during an acute exacerbation (preferred method). However, in children with no wheezing (or other signs of airflow obstruction) on presentation, reversibility may be determined by convincing parental report of a symptomatic response to a three-month therapeutic trial of a medium dose of inhaled corticosteroids with as-needed SABA (alternative method), or as-needed SABA alone (weaker alternative method). The authors provide key messages regarding in whom to consider the diagnosis, terms to be abandoned, when to refer to an asthma specialist and the initial management strategy. Finally, dissemination plans and priority areas for research are identified.     

The effect of lithium on growth factor production in long-term bone marrow cultures

We have previously reported that lithium chloride (LiCl) stimulates the production of granulocyte-macrophage colony-forming cells (GM-CFC), pluripotent stem cells (CFU-S), and differentiated granulocytes, macrophages and megakaryocytes in murine Dexter marrow cultures and that this effect appears to be mediated indirectly by a radioresistant adherent marrow cell. In this study we have established that exposure of murine Dexter cultures to LiCl (4 mEq/L) causes an increase of colony-forming cell megakaryocytes (CFU-meg) over 1 to 6 weeks of culture in both supernatant (188% to 611%) and stromal phases (123% to 246%). Moreover, we have shown that lithium treatment of either irradiated (1,100 rad) or unirradiated stromal cells increased production of activities stimulating formation of megakaryocyte, granulocyte, macrophage, and mixed lineage colonies and proliferation of the factor-dependent cell line, FDC-P1. This FDC-P1 stimulatory activity was completely blocked by an antibody to purified recombinant granulocyte-macrophage colony stimulating factor (rGM-CSF). The baseline or lithium-induced--stromal-derived bone marrow colony stimulating activity was partially blocked by the antibody to rGM-CSF and by an antibody to purified colony stimulating factor I (CSF-1); the two antibodies combined resulted in greater than 90% inhibition of the lithium-induced marrow stimulatory activity. In addition, radioimmunoassay (RIA) showed that although CSF-1 was detectable in supernatants of these cultures, exposure to lithium did not increase CSF-1 levels. These data indicate that Dexter stromal cells produce CSF- 1 and GM-CSF and that lithium appears to exert its stimulatory effects on in vitro myelopoiesis by inducing production of GM-CSF.     

Quantitative thallium-201 myocardial imaging in assessing right ventricular pressure in patients with congenital heart defects.

Thallium-201 myocardial scintigraphy was performed in patients with congenital heart defects to determine whether, by quantification of right ventricular isotope uptake, one could assess the degree of right ventricular hypertrophy and so predict the level of right ventricular pressure. A total of 24 patients ranging in age from 7 months to 30 years was studied; 18 were studied before corrective surgery and six after operation. All but three had congenital heart defects which had resulted in pressure and/or volume-overload of the right ventricle. At routine cardiac catheterisation, 20 microCi/kg thallium-201 as thallous chloride was injected through the venous catheter and myocardial images were recorded in anterior and left anterior oblique projections; these were subsequently analysed quantitatively and qualitatively. Insignificant right ventricular thallium-201 counts judged as being less than 1 per cent of the injected dose or less than 0.3 of the left ventricular counts were present in six patients all with right ventricular peak systolic pressure less than 30 mmHg. In the remaining 18 patients there was a good correlation between the right ventricular/left ventricular peak systolic pressure ratio and the right ventricular/left ventricular thallium-201 counts ratio. All patients with right ventricular/left ventricular peak systolic pressure less than 0.5 had right ventricular/left ventricular thallium-201 counts less than 0.4. Qualitative evaluation of right ventricular isotope intensity proved helpful mainly in distinguishing the patients with right ventricular pressures at or above systemic levels. Thus quantitative analysis of myocardial imaging with thallium-201 is of use clinically in patients with congenital heart defects, in assessing the severity of pulmonary stenosis or the presence of pulmonary artery hypertension.     

急性阑尾炎多层螺旋CT扫描征象与血清炎性标志物的相关性研究

目的探讨急性阑尾炎多层螺旋CT征象与血清炎性标志物的关系。方法收集2012年1月至2013年12月于同济大学附属杨浦医院经手术病理证实的具有完整临床及影像资料的急性阑尾炎患者66例,对所有患者的急性阑尾病变程度行CT分级,分析其与患者白细胞(WBC)计数、中性粒细胞百分比(NEUT%)及血清C反应蛋白(CRP)水平的相关性。结果急性阑尾炎CT分级与患者WBC计数与CRP水平呈正相关(P0.05),穿孔性阑尾炎患者NEUT%及CRP水平明显高于其他患者。CRP水平与阑尾直径、阑尾积液、回盲部变化、阑尾周围炎性条纹、小肠积液呈正相关(P0.05),WBC计数与回盲部变化及阑尾周围炎性条纹呈正相关(P0.05)。结论WBC计数及CRP水平与急性阑尾炎CT分级有关,CRP对急性阑尾炎的诊断及其严重程度的判断更有优势;CRP、NEUT%是穿孔性阑尾炎的重要预测因子,WBC可以更好地发现早期阑尾周围的炎性反应,综合分析CT表现与血清炎性标志物能更准确地诊断急性阑尾炎。     

纳米羟基磷灰石/硫酸钙复合人工骨顺铂缓释系统注射液的制备与研究

目的研制可注射α-CSH-nano-HA/PHBV-PEG顺铂释药系统,为骨转移瘤提供新型的局部药物缓释系统。方法α-CSH-nano-HA/PHBV-PEG载顺铂制成可注射用α-CSH-nano-HA/PHBV-PEG cis-platinum缓释微球,研究其结构、释药特性、可注射性以及力学性能。结果(1)第1、3、5、7天缓释微球的释药浓度分别为97.5、90.7、83.2、68.5μg/mL,第7天后趋于稳定。(2)可注射α-CSH-nano-HA/PHBV-PEG顺铂释药系统在液固比为0.7时可注射性强,与此同时缓释药系统随着液固比的增大凝固时间延长。结论α-CSH-nano-HA/PHBVPEG顺铂释药系统具有良好的注射性能和缓释作用。