CTLA4-Ig和抗CD40L单克隆抗体抑制大鼠胰腺移植急性排斥反应的机制

目的探讨细胞毒T淋巴细胞相关抗原4免疫球蛋白（CTLA4-Ig）和抗CD40L单克隆抗体对异基因大鼠胰腺移植后急性排斥反应的作用及其相关机制。方法建立大鼠的胰十二指肠移植模型，供者为F344大鼠，受者为经链尿佐菌素诱导为糖尿病模型的Lewis大鼠，受者移植后分为4组，每组12只。A组：为应用生理盐水对照组；B组：应用CTLA4-Ig200μg；C组：应用抗CD40L单克隆抗体200μg；D组：联合应用CTLA4-Ig和抗CD40L单克隆抗体各200μg。各组分别于术后第2d腹腔注射相应的药物。术后1、4、7、10d分别取各组的移植胰腺，进行常规病理检测；采用逆转录聚合酶链（RT-PCR）法检测移植物白细胞介素2（IL-2）、白细胞介素4（IL-4）、白细胞介素10（IL-10）、γ干扰素（IFN-γ）的表达；术后第1、4、7、10d取受者外周血，采用流式细胞术计数T细胞亚群CD3^＋、CD4^＋和CD8^＋；术后第4d取移植胰计数CD4^＋CD25^＋T细胞。结果病理检测显示：与A组相比，B、C组排斥反应明显减弱，D组几乎未发生排斥反应；B、C、D组IL-2的表达高峰延迟，且表达水平较A组有不同程度的降低，D组又较B、C组表达水平下降，差异有统计学意义；B、C、D组IFN-γ的表达水平较A组有不同程度的降低，但D组与B、C组的差异不显著；B、C组IL-4的表达水平较A组有不同程度的升高，D组较A、B、C组表达水平下降，差异有统计学意义；B、C组IL-10的表达水平较A组有不同程度的升高，D组与A组差异不显著；B、C、D组CD3^＋、CD4^＋和CD8^＋T细胞数均相对减少，CD4^＋CD25^＋T细胞数有不同程度升高，与A组比较，D组的差异最为显著。结论联合应用CTLA4-Ig和抗CD40L单克隆抗体能更有效地抑制大鼠胰腺移植后排斥反应，其机制可能与Th1／Th2型细胞因子偏移及CD4^＋CD25^＋调节性T细胞增多等有关。     

免疫分选软骨前体细胞并诱导永生化的研究

目的建立永生化大鼠软骨前体细胞株,为细胞移植和转基因治疗提供稳定的细胞来源。方法利用免疫磁珠技术分离纯化具有特异性表面标志成纤维生长因子受体-3(FGFR-3)的软骨前体细胞,用基因转染技术将含有猿肾病毒40大T抗原基因(SV40Tag)的重组质粒pEGFP- IRES2-SV40Tag转染原代培养的新生大鼠软骨前体细胞,经G418筛选,抗性克隆扩大培养。应用FGFR-3、Ⅱ型胶原和X型胶原抗体进行细胞鉴定,检测其分化能力,观察细胞的形态及其生长状况,绘制生长曲线。用逆转录-聚合酶链反应(RT-PCR)、Southern blot和免疫细胞化学法鉴定SV40Tag在转染细胞中的表达。结果获得1个阳性细胞克隆,免疫细胞化学证实为FGFR-3阳性的具有较强增值能力和多分化潜能的软骨前体细胞。经Southern印迹杂交证实,SV40Tag已稳定转染入软骨前体细胞,表达mRNA及其蛋白。贴壁培养的永生化软骨前体细胞株(IPSC),群体倍增时间为23.62 h,传代、冻存和复苏对细胞形态及生长无明显影响。结论SV40Tag导入可诱导软骨前体细胞永生化,为软骨前体细胞的实验研究及其介导的细胞移植治疗提供了稳定的细胞来源。     

构建含有小鼠CD40-IgG2aFc-IRS-GFP融合基因腺病毒并体外检测

目的 构建载有小鼠CD40-IgG2aFc-IRS-GFP融合基因片段的腺病毒载体,并检测其转染293细胞后的融合蛋白表达及出毒情况.方法 提取小鼠CD40、IgG2aFc基因片段构建PDC316-IgG2aFc-GFP质粒,测序成功后包装构建Ad5-PDC316-CD40-IgG2aFc-GFP,转染293细胞.出毒后大量复制病毒并纯化,测定病毒滴度,体外感染细胞观察病毒复制及分泌目的 蛋白情况并检测目的 蛋白表达量.结果 成功构建了质粒及病毒,体外感染显示该病毒能有效的感染细胞并表达蛋白(荧光显示),检测目的 蛋白表达量随时间点及浓度增加而增加.当病毒浓度增加到一定程度时目的 蛋白表达趋于无明显差异性.结论 构建小鼠融合基因片段并成功转入细胞内,分泌表达目的 蛋白是可行的,为进一步研究该病毒在大鼠体内感染及表达CD40Ig、大鼠肝移植模型免疫耐受及其机制的研究奠定了基础.     

Breast cancer in young women: Imaging and clinical course

The purpose of this study was to characterize presenting imaging findings in women younger than 40 diagnosed with invasive breast cancer in the context of pathology and clinical course. Retrospective chart and imaging reviews were performed in patients under 40 diagnosed with breast cancer between July 1, 2004, and December 31, 2013. Patient demographic, imaging, pathology, and clinical data were collected. Overall and recurrence-free survival were estimated using the Kaplan-Meier method. Univariate Cox proportional hazards models were performed to identify factors associated with recurrence-free survival. Our study cohort consisted of 110 patients with invasive mammary carcinoma. One hundred one (91.8%) presented with a palpable mass. The mean size of all lesions on imaging was 3.5 cm ± 2.9 cm. Malignant calcifications were present in 54 (49.1%) cases. Imaging demonstrated multifocal or multicentric disease in 45 (40.9%) cases. Seventy four (67.3%) cancers were high grade. Luminal genomic subtypes were the most common (n = 61, 55.5%). At presentation, 4 (3.6%) patients had bilateral malignancy and 8 (7.3%) patients had distant metastatic disease. Ninety seven (88.2%) underwent neoadjuvant chemotherapy and 67 (60.9%) underwent radiation therapy. Seventy five (68.2%) of the patients underwent mastectomy. The restricted mean time to recurrence was 9.01 years (standard error 3.162 months). ER positivity was associated with compromised recurrence-free survival. The overall survival rate was 0.962 at 10 years. Young patients diagnosed with breast cancer typically present with advanced breast imaging findings and undergo aggressive treatment. Recurrence often occurs >5 years from diagnosis, and ER positive subtypes are at increased risk for recurrence.     

A C >T polymorphism located at position −1 of the Kozak sequence of CD40 gene is associated with low bone mass in Spanish postmenopausal women

SUMMARY: This study evaluated the association of a polymorphism in the CD40 gene with BMD and risk of osteopenia or osteoporosis in a population of 602 postmenopausal women. Results showed that women with the TT genotype had lower BMD at femoral neck and spine sites and increased risk of osteopenia or osteoporosis. INTRODUCTION: Recent findings have demonstrated that the CD40/CD40L system, which is of main importance for the immune system, can also be implied in the regulation of bone metabolism. The main objective of the present work has been to clarify whether single nucleotide polymorphisms (SNPs) affecting genes of CD40/CD40L system could be linked with abnormalities in the level of bone mineral density (BMD) in menopausal women. METHODS: We performed an association study of BMD values with a SNP located at position -1 of the Kozak consensus sequence of CD40 gene (rs1883832; C>T) in a population of 602 postmenopausal women. RESULTS: Women with the TT genotype (8.6% of women) displayed a reduction in femoral neck BMD (FN BMD) and lumbar spine BMD (LS BMD) of 6.2% and of 6.3%, respectively, as compared to women with CC + CT genotype. Logistic regression analysis adjusted for age, weight, and height showed that women with the TT genotype had increased risk for FN (odds ratio: 2.34; 95% CI: 1.12-4.89) and LS (odds ratio: 2.49; 95% CI: 1.19-5.24) osteopenia or osteoporosis. CONCLUSIONS: Women with the TT genotype in rs1883832 SNP affecting to Kozak consensus sequence of CD40 gene had lower BMD at FN and at LS sites and increased risk of osteopenia or osteoporosis.     

Rosiglitazone down-regulates lipopolysaccharide-induced expression of CD40 and intercellular adhesion molecule 1 in rat peritoneal mesothelial cells through a NF-κB dependent mechanism

Objective To investigate the effect and mechanism by which PPARγ ligand, rosiglitasone, regulates the expression of CD40 and intercellular adhesion molecule 1 (ICAM-1) in the rat peritoneal mesothelial cells (RPMCs) induced by lipopolysaccharide (LPS). Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were randomly divided into groups as follows: medium, LPS (5 mg/L), LPS (5 mg/L)+BAY11-7085(5 μmol/L, NF-κB inhibitor), rosiglitazone (10 μmol/L or 20 μmol/L, peroxisome proliferator-activated receptor γ activator), LPS (5 mg/L)+rosiglitazone (10 μmol/L)+GW9662 (3 μmol/L, peroxisome proliferator-aetivatcd receptor γ antagonist), and LPS (5 mg/L)+vehicle (DMSO 0.2 ml/L). The expressions of CD40 and ICAM-1 RNA in RPMCs were examined by RT-PCR after 3 hour treatment, and the protein expressions of CD40, ICAM-1, p-NF-κB p65 and p-IκBα were examined by Western blot or immunofluorescence after 24 hour treatment. Results Following treatment with LPS, both the expressions of CD40 and ICAM-1 protein in RPMCs were up-regulated significantly (P<0.05), and the phosphoralation of p65 was increased greatly (1.10±0.17 vs 0.55±0.06, P<0.05). BAY11-7085 (5 μmol/L) significantly decreased the protein expression of p-p65 (0.22±0.11 vs 1.10±0.17, P<0.01), CD40 (0.34±0.02 vs 0.50±0.06, P<0.05) and ICAM-1 (0.35±0.16 vs 0.74±0.03, P<0.05). Pretreated with rosiglitazone for 3 h then added with LPS for 1 h, the levels of p-p65, CD40 and ICAM-1 in RPMCs were significantly decreased compared with those of LPS group (0.77±0.08 vs 0.90±0.10, P相似文献   

Mutations in the transcription factor FOXO1 mimic positive selection signals to promote germinal center B cell expansion and lymphomagenesis

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Echocardiographic criteria for aortic root dissection.

Echocardiographic findings from 10 patients without clinical indications of aortic root dissection or aortic valve disease from 1 patient with angiographic confirmation of aortic root dissection are reported and compared. Previously reported echocardiographic findings were confirmed in the patient with aortic root dissection. These include (1) a widened posterior or anterior aortic wall, or both; (2) parallel motion of the separated margins of the aortic walls; and (3) aortic root dilatation (42 mm or more at end-systole). However, all three findings were also noted in 5 of the 10 patients without clinical indications of aortic root dissection or aortic valve disease, and at least two of the three findings were noted in the remaining 5 patients. Echocardiographic detection of aortic root dissection appears to be most reliable when clinical indications of the anomaly are present.     

The use of dextran-40 during percutaneous transluminal coronary angioplasty: A report of three cases of anaphylactoid reactions—one near fatal

Successful percutaneous transluminal coronary angioplasty is achieved by fracture of the atheromatous plaque and perhaps dilatation of the arterial walls to increase the luminal diameter of the artery. Because this ?controlled”? injury stimulates platelet adhesion on the subendothelial matrix, the use of dextran-40 in addition to heparin has been advocated. The overall incidence of Dextran-induced anaphylactoid reactions at our institution was 0.6% and of severe life-threatening reactions, 0.2%. With the recent doubts cast on the efficacy of dextran-40, the question arises regarding its routine use in PTCA.     

Simian virus (SV) 40 like sequences in cell lines and tumour biopsies from Australian malignant mesotheliomas

Background : Simian virus (SV) 40 sequences have been found in some, but not all studies of mesotheliomas. This virus is known to cause tumours in rodents but its role in human oncogenesis remains controversial.
Aims : The aim of this study therefore was to determine whether SV40 is associated with the development of mesotheliomas in Australia. The absence of the virus or its gene products in tissue derived from mesotheliomas would detract from this possibility.
Methods : We used polymerase chain reaction from three pairs of primers to amplify different regions of the large T antigen from DNA from cell lines and cDNA from both cell lines and an independent set of tumour biopsies from patients with mesothelioma.
Results : We examined five human mesothelioma cell lines that were established in our laboratories. In addition, we examined several tumour biopsies from seven different patients. SV40 like sequences were present in all the cell lines and in at least one sample from each of the patients examined.
Conclusions : The large T antigen of SV40 or an SV40 like virus is expressed in Australian mesotheliomas and therefore could be aetiologically-associated with tumourigenesis. Alternatively, these sequences could be expressed subsequent to the development of the disease.