Determination of cardiac troponin I forms in the blood of patients with unstable angina pectoris

OBJECTIVE: To determine the predominant form in which cardiac troponin I circulates in the bloodstream of unstable angina patients. DESIGN AND METHODS: The cardiac troponin I forms released in the bloodstream of 25 patients suffering from unstable angina were examined by using three immunoenzymatic assays: the total cTnI assay for detection of free and complexed cTnI, the IC-TIC assay for detection of the IC and TIC troponin complexes, and the IT-TIC assay for detection of the IT and TIC troponin complexes. RESULTS: Approximately 60% of patients with unstable angina had at least one positive value in the total cTnI assay or in the IC-TIC assay. Our results demonstrated that the predominant cardiac troponin I form circulating in the bloodstream of patients with unstable angina was the IC complex. Free cTnI, IT, and/or TIC forms were seldom found, the frequency of IT and/or TIC complexes being higher than that observed previously in patients with acute myocardial infarction. CONCLUSIONS: The release pattern of cTnI in patients suffering from unstable angina is similar to that previously observed in patients with acute myocardial infarction, i.e., a predominance of the IC complex.     

C6细胞中诱导体外培养大鼠神经干细胞迁移蛋白的初步分析

目的：许多体内实验表明神经干细胞具有向胶质瘤细胞迁移的特性，该特性使神经干细胞有可能成为基因治疗脑胶质瘤潜在的基因携带者。实验拟观察大鼠星形胶质瘤C6细胞诱导体外培养的大鼠神经于细胞的迁移作用，并初步分离C6细胞中诱导神经干细胞迁移的蛋白： 方法：实验于2005—01/2006—05在南通大学江苏省神经再生重点实验室完成。实验材料：C6细胞用含10％小牛血清的DMEM/F12培养基进行培养，使用时细胞处于对数生长期：清洁级的新生1周内的SD大鼠由南通大学实验动物中心提供，实验过程中对动物处置符合动物伦理学标准。实验方法：从新生5～7dSD大鼠大脑皮质分离培养、纯化神经干细胞。从SD新生红皮鼠大脑皮质分离、培养、纯化星形胶质细胞。采用“Transwell Inserts”细胞培养室培养系统共培养36h，比较C6细胞和大鼠星形胶质细胞对体外培养的大鼠神经干细胞的迁移作用；自然系统聚丙烯酰胺凝胶蛋白电泳分离C6细胞和星形胶质细胞差异蛋白，观察各蛋白凝胶条带对神经干细胞的迁移作用。 结果：体外培养C6细胞能诱导神经干细胞迁移，迁移细胞数目与星形胶质细胞相比，差异有显著意义（P〈0．001）。将C6细胞中差异蛋白条带与神经干细胞共培养，与对应的星形胶质细胞蛋白条带相比，可观察到与相对分子质量约67000蛋白胶条共培养的神经细胞球在接近蛋白胶条的一侧细胞向胶条迁移生长。 结论：C6细胞可以诱导体外培养的神经干细胞迁移，其总蛋白在自然系统聚丙烯酰胺凝胶电泳分离实验中获得的相对分子质量为67000的蛋白组分具有诱导神经干细胞迁移的活性。     

腰骶部慢性骨筋膜间隔综合征解剖及临床研究现状

目的:总结近年来腰骶部慢性骨筋膜间隔综合征的研究现状,以期提高人们对腰骶部骨筋膜间隔综合征的认识、诊断及治疗。方法:应用计算机检索CNKI1994-01/2007-03及万方数据库2000-01/2007-03相关腰骶部慢性骨筋膜间隔综合征方面的文献,检索词"下腰痛,腰肌劳损,腰骶部骨筋膜间隔综合征",限定文章语言种类为中文。同时计算机检索PubMed、Medline、Springerlink2000-01/2007-03及Embase数据库2003-01/2007-03相关腰骶部慢性骨筋膜间隔综合征方面的文献,检索词"LBP,lumbar muscle strain,Chronic Compartment syndrome",并限定文章语言种类为English。对资料进行初审,选取包括腰骶部慢性骨筋膜间隔综合征的文献,开始查找全文,并查看每篇文献后的引文。纳入标准:文章所述内容应与慢性骨筋膜室综合征、慢性下腰痛、肌内压测定有关。排除标准:重复研究或Meta分析类文章。结果:共检索到45篇关于腰骶部慢性骨筋膜间隔综合征的文献,最终纳入15篇符合标准的文献。腰骶部骨筋膜间隔综合征目前尚未被普遍认识,但是1981年Peck就首次报道腰部骨筋膜间隔综合征,并提出该病可能是下腰痛原因之一。腰骶部骨筋膜间隔综合征与其他部位的骨筋膜间隔综合征病理变化相似,若得不到及时治疗就会引起肌肉的变性、坏死及纤维化,这是腰肌劳损形成的重要病理原因之一。诊断腰骶部慢性骨筋膜间隔综合征需要在排除其他疾病的基础上结合骨筋膜间隔内压力测定。骨筋膜间隔内压力的直接测定仍然是骨筋膜间隔综合征诊断的金标准。目前骨筋膜间隔综合征治疗四肢部位都趋向于行解压手术治疗,腰骶部的骨筋膜间隔综合征治疗意见不一,保守治疗虽然可以暂时缓解酸痛症状但疗效没有确切的统计学依据,并且不能预防阻止受压的骨骼肌变性。筋膜切开解压术是针对此病的基本疗法,有关专家的研究表明术前术后效果明显。因此对腰骶部骨筋膜间隔综合征的认识、诊断及治疗,对预防或减少慢性腰肌劳损所致的慢性腰痛具有重要意义。结论:腰骶部慢性骨筋膜间隔综合征可引起下腰痛,保守治疗虽然可以暂时起效但不能根本解决问题,手术治疗仍是基本的治疗手段。     

重组腺相关病毒介导增强型绿色荧光蛋白在骨髓间充质干细胞中的表达

目的:重组腺相关病毒是携带治疗目的基因的有效载体.骨髓间充质干细胞则可作为外源基因的载体和表达场所实验选择携带绿包荧光蛋白的腺相关病毒体外转染骨髓间充质干细胞,观察转染效果。方法:实验于2006-01/12在咸宁学院心血管疾病研究所和武汉大学心内科实验室完成清洁级成年SD大鼠6只.由武汉大学医学院试验动物中心提供,实验过程中对动物的处置符合动物伦理学标准。携带增强型绿色荧光蛋白的重组腺相关病毒载体由北京本元正阳基因技术公司提供,基因启动子为CMV,有效滴度4×10~(11)v.g/mL。全骨髓法分离大鼠骨髓间充质干细胞,于体积分数为0.1的胎牛血清中培养.扩增纯化至第3代。按感染复数分别为10~2.10~3.10~4,10~5 v.g/cell加入携带增强型绿色荧光蛋白的重组腺相关病毒。于4,8,24、48 h倒置相差荧光显微镜下观察腺相关病毒感染骨髓间充质干细胞的状态,随机记录200个细胞中增强型绿色荧光蛋白阳性表达率.计算感染效率。结果:①骨髓间充质干细胞转染效率:感染复数为10~3.10~4,10~5 v.g/cell时.转染8 h后骨髓间充质干细胞内可见绿色荧光蛋白的表达.其中105 v.g/cell感染效果最佳.于转染48 h时荧光强度达最高值.感染效率约28%②骨髓间充质干细胞经G418筛选后检测增强型绿色荧光蛋白阳性细胞率达98%。结论:介导绿色荧光蛋白的腺相关病毒转染骨髓间充质干细胞简便易行,感染复数为10~5v.g/cell时效果较佳,目的基因表达稳定,用于后续基因治疗具备一定的可行性。     

SD大鼠骨代谢与非洛地平的干预

目的:流行病学报告提示治疗心血管疾病的重要药物钙离子通道阻滞剂影响骨代谢,但这种作用与药物的剂量和作用时间的关系还不明确。观察不同剂量、不同时间的非洛地平对SD大鼠骨代谢的作用。方法:实验于2006-05/2007-04在西安交通大学动物实验中心完成。①实验分组:132只6月龄SD大鼠,随机分为4组:正常对照组,非洛地平0.15,0.45,1.25mg/kg组,每组33只。②实验方法:非洛地平各组每天按0.15mg/kg,0.45mg/kg,1.25mg/kg剂量灌胃非洛地平,正常对照组灌服等量生理盐水。③实验评估:在实验第3,4,5,6个月时每组随机取8只大鼠,对其股骨和腰椎骨密度、身长、体质量和血清游离钙、磷、碱性磷酸酶、胆固醇、三酰甘油含量进行测定。结果:进入结果分析的SD大鼠共有127只,正常对照组31只,非洛地平0.15mg/kg组31只,非洛地平0.45mg/kg组32只,非洛地平1.25mg/kg组33只。①各组大鼠股骨和腰椎骨密度:给药后6个月后非洛地平0.15mg/kg组大鼠的股骨和腰椎骨密度高于正常对照组(P<0.01),非洛地平0.45,1.25mg/kg组股骨骨密度则出现降低(P<0.01),非洛地平1.25mg/kg组降低更明显(P<0.01)。②各组大鼠的身长、体质量和血清钙、磷、碱性磷酸酶、胆固醇、三酰甘油含量:不同剂量的非洛地平组与正常对照组比较,差异无显著性。各时间段比较差异也无显著性。结论:①短期应用非洛地平对大鼠骨代谢无明显影响。②长期应用低剂量非洛地平对大鼠股骨和腰椎的骨代谢产生正性的影响。③长期应用中、高剂量非洛地平对大鼠负重骨(股骨)的骨代谢产生负性的影响,剂量越大,负性影响越明显;而对非负重骨(腰椎)的骨代谢无明显影响。     

SLE-ASSOCIATED THROMBOCYTOPENIA IN PREGNANCY: SUCCESSFUL OUTCOME WITH COMBINED IgG INFUSION AND PLATELET TRANSFUSION

SUMMARY Autoimmune thrombocytopenia presents special problems during pregnancy because of the limited therapeutic options and the risks posed by even moderate degrees of thrombocytopenia. Although corticosteroids and intravenous immunoglobulin G (IV IgG) have been shown to be useful and relatively safe therapies, they are not always effective. We describe a patient with systemic lupus erythematosus (SLE) who developed severe thrombocytopenia unresponsive to prednisone and IV IgG. The combination of IV IgG and platelet transfusion immediately before delivery resulted in a rapid rise in platelet count and allowed a lower segment caesarean section to be undertaken without perioperative complication.     

Hairy cell identification by immunohistochemistry of tartrate-resistant acid phosphatase

Tartrate-resistant acid phosphatase (TRAcP) has been an indispensible marker for hairy cell leukemia (HCL) for over two decades. However, the traditional TRAcP cytochemical stain cannot be performed effectively on sections of paraffin-embedded tissues that are important resources for histopathologic evaluation in diagnosis and treatment of HCL. Wide variation in expression of TRAcP activity by hairy cells (HCs) within and among patients is an interesting biologic phenomenon that has not been explained and can cause some diagnostic uncertainty as well. To solve the problem of staining TRAcP in paraffin sections and to begin to address the questions of variable TRAcP expression in HCL, we developed a monoclonal antibody to TRAcP, 9C5, for immunohistochemical identification of HCs. In smears of blood and bone marrow, immunocytochemistry of TRAcP using 9C5 was as specific but slightly less sensitive than direct cytochemical staining of enzymatic activity. In paraffin sections of spleen and bone marrow from HCL patients, immunohistochemistry with 9C5 stained the HCs with high sensitivity and specificity and clearly showed the characteristic diffuse infiltration by HCs. Other cells noted to stain strongly with 9C5 were occasional macrophages in bone marrow smears and osteoclasts and occasional tissue macrophages in paraffin sections. These are cells known to express abundant TRAcP activity. Immunohistochemistry with anti-TRAcP monoclonal antibody 9C5 may have utility as an added option in the diagnosis of HCL, as a means to evaluate residual disease in HCL patients undergoing new treatments, and as a way to address questions regarding variable expression of TRAcP activity by HCs within and among patients with HCL. Also, 9C5 has potential as a reagent for the immunoassay of bone-derived serum TRAcP in patients with certain bone diseases and cancers with bone metastasis.     

Factors influencing platelet transfusion refractoriness in patients undergoing allogeneic hematopoietic stem cell transplantation

Hematopoietic stem cell transplantation has been considered a risk factor for development of platelet transfusion refractoriness. The objective of this study was to assess the platelet transfusion refractoriness rate in patients undergoing allogeneic hematopoietic stem cell transplantation from different sources. We retrospectively reviewed the charts and transfusion records of patients who underwent allogeneic stem cell transplantation at our institution between 2013 and 2015. The evaluation of post-transfusion platelet count was assessed for each transfusion given, from day of progenitor infusion to day 30 after transplantation. Of 167 patients included in this study, 101 received peripheral blood stem cell transplantation (PBSCT) and 66 received umbilical cord blood transplantation (UCBT). Overall, the percentage of platelet transfusions with a 14-h CCI lower than 5000 was 59.3%, being these data significantly higher for UCBT (67.6%) than for PBSCT (31.0%). Seventy-eight percent of patients underwent UCBT become refractory, while 38.6% of patients who received PBSCT were refractory. Factors associated to platelet refractoriness were lower CD34+ cell dose infused, higher number of antibiotics used, presence of anti-HLA I antibodies, and reduced-intensity conditioning regimen. Platelet refractoriness is a frequent and complex adverse event and remains a therapeutic challenge in the management of patients undergoing HSCT. There is a higher rate of platelet refractoriness in patients who received UCBT as compared to patients who received PBSCT.     

Demographic and immune correlates of human herpesvirus 8 seropositivity in Malawi, Africa.

BACKGROUND: In the USA, human herpesvirus 8 (HHV-8) is associated with Kaposi's sarcoma (KS) and HIV infection. We examined HHV-8 seroprevalence in a Malawian cohort, and assessed its relationship with HIV, KS, demographic characteristics, and immune findings. METHODS: In 1997 and 1998, blood samples were obtained from 272 hospitalized Malawian patients, for whom demographic information was obtained, and 24 healthy volunteers without demographic data. We used enzyme immunoassays to assess seroprevalence and antibody titers to peptide antigens derived from HHV-8 K8.1 and ORF65-encoded proteins. Intracellular cytokines and cell surface antigens were assessed with four-color flow cytometry. Data were analyzed using non-parametric univariate and regression analytic techniques. RESULTS: The rates of HHV-8 seroprevalence to either or both HHV-8 peptides were 67% for the patients and 54% for the healthy volunteers. Seroprevalence increased with patients' age (P<0.001) but was not associated with HIV status, percentage of lymphocytes expressing CD4, or KS (n=10). Seropositive females had lower antibody titers to both peptides than did males (medians: 455 versus 1361 for K8.1, P<0.001; and 268 versus 405 for ORF65, P=0.044). For the healthy volunteers, the percentage of CD8+ cells producing IFN-gamma after stimulation was significantly lower in ORF65-specific antibody-positive persons (medians: 24% versus 57%, P=0.008). CONCLUSIONS: In Malawi, HHV-8 is endemic and is not associated with HIV infection or HIV severity. Seroprevalence rates increase in childhood, and, most steeply in adolescence. Titers are higher in seropositive males than in sero-positive females. The immune effects of HHV-8 in healthy adults are consistent with chronic inhibition of type 1 cytotoxic T-cell responsiveness, independent of HIV status.