2型糖尿病患者外周血CD8+CD25+Foxp3+Treg细胞检测及意义

目的探讨外周血CD8~+CD25~+Foxp3~+Treg(CD8~+Treg)细胞比例在2型糖尿病(T2DM)疾病过程中的作用。方法选取T2DM患者90例。根据尿微量白蛋白(MAL),分为45例T2DM伴正常蛋白尿患者(NMAU,MAL30 mg/24 h),45例T2DM伴微量蛋白尿患者(MAU,MAL 30～300 mg/24 h),同时选择门诊体检健康人员45名作为对照组(HC)。通过特定蛋白仪检测患者mAlb浓度;流式细胞仪检测CD8~+Treg细胞比例;Luminex200检测血浆白细胞介素(IL)-8、1β、6和肿瘤坏死因子(TNF)-α浓度。结果与HC组[4.43(3.26～5.75)%]相比,NMAU组[4.06(3.15～5.43)%]和MAU组[3.52(2.30～4.87)%]患者CD8~+Treg细胞比例降低,且MAU组降低更明显(P0.05);与HC组相比,MAU组和NMAU组患者血浆细胞因子TNF-α、IL-8、IL-6和IL-1β浓度均升高(P0.05);MAU组患者CD8~+Treg比例与血清细胞因子TNF-α(r=-0.444 8,P0.01)、IL-8(r=-0.341 2,P0.05)、IL-6(r=-0.365 3,P0.05)和IL-1β(r=-0.319 2,P0.05)呈负相关;Logistic回归分析显示CD8~+Treg对T2DM患者可能具有保护作用。结论通过调控T2DM患者外周血CD8~+Treg细胞比例,有可能减缓糖尿病的进程。     

2型糖尿病患者尿Ⅳ型胶原和尿微量白蛋白联合测定的意义

为评价尿微量白蛋白与Ⅳ型胶原对2型糖尿病患者肾功能早期损伤的诊断价值, 取正常人30名与165例2型糖尿病患者24小时的尿液, 采用放射免疫法(RIA), 同时测定尿微量白蛋白和Ⅳ型胶原(浓缩法).结果显示: (1)正常组24h尿微量白蛋白为6.45±2.04mg/24h,尿Ⅳ型胶原为86.68±11.98μg/L.(2)根据2型糖尿病患者24h尿微量白蛋白含量分为两组,A组130例, 尿微量白蛋白大于或等于30mg/24h,尿Ⅳ型胶原为120.03±34.02μg/L; B组35例, 尿微量白蛋白小于30mg/24h, 尿Ⅳ型胶原为97.14±24.93μg/L.结论: 2型糖尿病患者在尿微量白蛋白还未超过正常上限时, 已经有部分人尿Ⅳ型胶原升高, 说明尿Ⅳ型胶原比尿微量白蛋白更能体现2型糖尿病患者肾功能早期损伤.     

2型糖尿病患者微量白蛋白尿的相关危险因素分析

目的:研究2型糖尿病(NIDDM)患者微量白蛋白尿(MAU)的危险因素.方法:用放射免疫法测定了112例NIDDM尿白蛋白排泄率(UAER),利用多因素逐步回归分析法分析MAU的危险因素,并显示这些因素独立作用.结果:NIDDM患者UAER与胆固醇、舒张压、空腹血糖、体重指数呈独立正相关;性别、年龄、高密度脂蛋白无明显作用.结论:高血脂、高血压、高血糖、肥胖将导致NIDDM患者发生MAU.     

2型糖尿病患者颈动脉粥样硬化与尿微量白蛋白的关系

目的探讨2型糖尿病患者颈动脉硬化与尿微量白蛋白的关系。方法收集109例2005年至2008年在本院住院的2型糖尿病患者,采用彩色多普勒超声诊断仪10MHz高频探头检测患者双侧颈动脉,按超声检查结果分为4组：颈动脉正常组30例[双侧颈动脉内膜中层厚度（CIMT）〈0．9mm];颈动脉内膜增厚组30例（0．9mm≤CIMT≤1．3mm）;稳定斑块组23例（斑块呈强回声,伴或不伴声影）;不稳定斑块组26例（斑块呈略低回声,内部或周边／有强回声附着）。同时测定血肌酐、空腹血糖（FPG）、糖化血红蛋白（HbA1c）、血清总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL）、高密度脂蛋白胆固醇（HDL）、尿微量白蛋白（MAU）,记录血压并计算体质量指数,比较四组患者MAU的差异。结果各组年龄、性别、体质量指数、血压、血肌酐比较差异无统计学意义（P〉0．05）,MAU值各组与CIMT〈0．9mm组间差异有显著性（P〈0．01）,尤其是不稳定斑块组与正常组差异最显著（P〈0．01）,不稳定斑块组与稳定斑块组、内膜增厚组,稳定斑块组与内膜增厚组差异也有统计学意义（P〈0．05）,不稳定斑块组与正常组在病程、FPG、Hba1C、TC、TG、LDL、HDL比较差异均有统计学意义（P〈0．05）,稳定斑块组与内膜增厚组除TC、MAU差异有统计学意义外（P〈0．05）,其余指标差异均无统计学意义（P〉0．05）。结论2型糖尿病患者颈动脉硬化程度与尿微量白蛋白水平呈正相关,MAU可预测2型糖尿病患者心血管疾病的风险。     

黄芪注射液对2型糖尿病尿微量白蛋白的影响

目的 探讨黄芪注射液对2型糖尿病尿微量白蛋白的影响.方法 将我院分2型糖尿病患者68例,其中男38例、女30例,平均年龄58岁.均符合1998 年WHO 糖尿病诊断标准.随机分为治疗组34例和对照组34例,治疗组采用在常规治疗的基础上加用黄芪注射液20 ml,加入0.9%氯化钠注射液250 ml中静脉滴注,1次/d两周为一疗程.对照组给予生理盐水500m l,每天1 次,静脉滴注2 周为一疗程.严格测定治疗前后两组患者在休息状态下卧位右侧肱动脉压,空腹静脉血浆葡萄糖(FPG),及连续2d24h尿微量白蛋白(UA E) 平均值.结果 对照组治疗前后UA E 无明显变化,而治疗组(黄芪注射液治疗)治疗2 周后较治疗前UA E 显著下降(P < 0.01),与对照组比较有显著性差异(P < 0.01).结论 黄芪注射液使2型糖尿病患者尿微量白蛋白明显下降.     

2型糖尿病患者血内皮素和尿微量白蛋白对肾功能受损的初步探讨

糖尿病是一种较常见的内分泌代谢障碍性疾病 ,特点是以胰岛素缺乏及糖代谢紊乱为主 ,继发脂肪、蛋白质代谢障碍、水、电介质丢失 ,以及急、慢性并发症的临床表现[1] ,主要病理改变是血管病变 ,其特征性的是微血管病变。常见的是肾脏、眼、神经系统、心血管、骨及皮肤的并发症 ,可以单一器官或多器官受累。本文报告 2型糖尿病患者血内皮素和尿微量白蛋白对肾功能受损程度的探讨 ,现报告如下。对象和方法一、对象 :(一 )正常人组 :35人 ,均为健康人 ,无心、肝、肺、肾等重要脏器疾患 ,肝、肾功能试验正常 ,无糖尿病史。(二 )病人组 :6 8人 ,均…     

2型糖尿病肾病患者外周血单个核细胞组织因子活性检测的意义

目的探讨2型糖尿病肾病患者外周血单个核细胞组织因子促凝活性（TF-PCA）的改变及其临床意义。方法 63名2型糖尿病患者按照尿白蛋白/肌酐比共分为三组：比值〈30mg/g为正常蛋白尿组（n=22）,30mg/g≤比值〈300mg/g为微量白蛋白尿组（n=26）,比值≥300mg/g为大量白蛋白尿组（n=15）;同时选择21名健康体检者作为对照组。采用一期凝固法测定外周血单个核细胞TF-PCA。常规检测空腹血糖、糖化血红蛋白、超敏CRP（Hs-CRP）、尿酸（UA）、胱抑素C（CYSC）,并进行相关分析。结果糖尿病组外周血TF-PCA随尿蛋白水平升高而升高,同时单个核细胞TF-PCA与空腹血糖、Hs-CRP、UA等呈正相关,相关系数分别为0.419、0.293、0.232（P〈0.05）。结论 2型糖尿病患者外周血单个核细胞TF-PCA明显升高,且与多种因素相关,TF-PCA升高可能与DN的发病机制、病程进展相关。     

2型糖尿病合并症患者血清唾液酸和尿微量白蛋白的检测分析

本文报道2型糖尿病合并症患者血清唾液酸，尿微量白蛋白等指标的检测分析结果。     

2型糖尿病检测晨尿Alb的临床价值

目的评价测定2型糖尿病患者晨尿白蛋白(Alb)的临床价值。方法同期检测20例正常人和33例2型糖尿病患者晨尿Alb/Cr比值、Alb和α1-微球蛋白(α1-MG)浓度。结果(1)Alb/Cr≥2.5mg/mmol为早期糖尿病肾病诊断标准 ,早期糖尿病肾病组晨尿Alb较糖尿病尿白蛋白正常组显著升高 (P<0.001) ,亦较正常人显著升高(P<0.001)。(2)晨尿Alb和尿Alb/Cr在正常人或2型糖尿病人中 ,均呈正相关。结论晨尿Alb可作为监测2型糖尿病肾脏早期病变指标之一。     

糖尿病肾病患者Ⅳ型胶原含量变化及其临床意义

目的探讨糖尿病肾病 (DN)患者Ⅳ型胶原 (Ⅳ -C)含量变化及其临床意义。方法57例2型糖尿病 (DM)患者根据尿白蛋白排泄率 (UAER)分为正常白蛋白尿组 (Ⅰ )、微量白蛋白尿组 (Ⅱ )和大量白蛋白尿组 (Ⅲ ) ,分别检测各组血、尿Ⅳ -C及尿微量白蛋白 (MA)、尿α1-微球蛋白 (α1-MG)含量 ,并与40例正常对照组 (NC)进行比较。结果DM患者血和尿Ⅳ -C含量均显著高于NC组 (P<0.01)。血Ⅳ -C、尿Ⅳ -C、UAER及α1-MG含量在NC组、Ⅰ组、Ⅱ组、Ⅲ组间呈逐组增高趋势 ,除血Ⅳ -C在Ⅰ组与NC组间无统计学差异外 (P>0.05),其余各组间结果均呈显著差异 (P<0.05)。尿Ⅳ -C与UAER、α1-MG呈显著正相关(P<0.01)。结论Ⅳ -C与DN的发生和发展有关。测定DM患者血、尿Ⅳ -C ,尤其是尿Ⅳ -C含量 ,不仅可作为诊断DN早期损害较灵敏的预警指标 ,还将有助于监测、判断DN的病程进展。     

慢性肾功能衰竭患者外周血细胞CD146的表达及意义

目的 探讨CD146在慢性肾功能衰竭（CRF）患者外周血细胞中的表达及意义.方法 收集本院2007年8月至2009年7月门诊及住院CRF患者51例为CRF组,33例正常人为健康对照组,流式细胞术检测两组外周血细胞CD146的表达并比较两组之间的差异,分析CRF患者外周血细胞CD146表达与血尿素氮（BUN）、血肌酐（Scr）的相关性.结果 与对照组比较,CRF组单核细胞、中性粒细胞CD146＋细胞百分率及CD45-CD146＋细胞数量均显著升高[（4.09±3.48）%比（0.20±0.10）%;（4.71±3.94）%比（0.79±0.14）%;（16.43±10.48）个/μl比（2.16±0.81）个/μl,均P〈0.001].CRF患者外周血CD45-CD146＋细胞数量与BUN、Scr均呈正相关（r=0.480、0.595,均P〈0.01）.结论 外周血单核细胞及中性粒细胞CD146的表达及升高与CRF的发生有关,CD45-CD146＋细胞数量增加与CRF的发生及严重程度有关.     

慢性肾功能衰竭患者外周血细胞CD146的表达及意义

Objective To study the clinical significance and expression of CD146 on peripheral blood cells in patients with chronic renal failure (CRF). Methods Expression of CD146 in peripheral blood samples from 51 ambulatory or hospitalized patients with CRF (registered between August 2007 and July 2009 to our hospital) and from 33 normal controls were determined and compared between two groups. Correlation of peripheral blood CD146 with blood urea nitrogen (BUN) , and serum creatinine (Scr) in patients with CRF was analyzed. Results Compared with normal controls, the rate of CD146* on monocyte and neutrophile in patients with CRF were significantly higher [ (4.09±3.48)% vs (0.20±0.10)% ;(4.71± 3.94)% vs (0.79±0.14)% , both P<0.001], and the number of CD45-CD146* cells in patients with CRF was obviously higher [ (16.43±10.48) /μl vs (2.16±0.81 )/μl,P<0.001) ]. The number of CD45-CD146+ cells in patients with CRF was positively correlated with BUN and Scr (r=0.480 and 0.595, both P<0.01). Conclusions Correlation between increased CD 146 on monocyte or neutrophile and the pathogenesis of CRF is found. The increase in CD45-CD146+ cell numbers is correlated with the pathogenesis and severity of CRF.     

The role of CD146 (Mel-CAM) in biology and pathology

CD146, also known as Mel-CAM, MUC18, A32 antigen, and S-Endo-1, is a membrane glycoprotein which functions as a Ca²⁺-independent cell adhesion molecule involved in heterophilic cell–cell interactions. Based on homology of the nucleotide sequence, CD146 is classified as a member of the immunoglobulin gene superfamily, since it contains the characteristic V-V-C2-C2-C2 immunoglobulin-like domain structure. Using immunohistochemistry with CD146-specific antibodies, CD146 expression has been demonstrated in a relatively limited spectrum of normal human tissues and malignant neoplasms. The lineage-specific expression pattern of CD146 can be useful in the differential diagnosis of certain lesions including melanomas and various types of gestational trophoblastic lesions. Although the biological role of CD146 in normal tissue and malignant tumours remains unclear, CD146 has been suggested to play an important role in tumour progression, implantation and placentation. CD146 expression can promote tumour progression in human melanoma, possibly through enhanced interaction between melanoma cells and endothelial cells. In contrast, CD146 may act as a tumour suppressor in breast carcinoma. CD146 expression is frequently lost in breast carcinomas and overexpression of CD146 in breast carcinoma cells results in a more cohesive cell growth and the formation of smaller tumours in nude mice. During implantation and placentation, CD146 expressed by the intermediate trophoblast in the placental site binds to its putative receptor in uterine smooth muscle cells and limits trophoblastic invasion in the myometrium. In conclusion, CD146 is a recently identified novel cell adhesion molecule and its biological functions and role as a diagnostic marker in pathology are now being recognized. Identification of the receptor for CD146 and the development of experimental models that can account for the complex interactions between CD146-expressing cells and their microenvironment are needed to investigate further the functions of this molecule in biology and in pathological states. Copyright © 1999 John Wiley & Sons, Ltd.     

CD146 protein in prostate cancer: revisited with two different antibodies

AIMS: CD146 is a potentially metastasis promoting cell adhesion molecule and its expression has been described in various solid tumours. We aimed to evaluate the expression of CD146 in prostate cancer by immunohistochemistry in a clinically characterised study cohort to evaluate its prognostic properties. METHODS: We evaluated the CD146 protein expression using a polyclonal and a monoclonal antibody on 169 clinico-pathologically characterised cases. Statistical analyses were applied to test for correlations and diagnostic and prognostic associations. RESULTS: CD146 detection with the polyclonal antibody revealed marked differential expression between tumour and normal tissue and was also a significant marker for shortened PSA relapse free survival. The monoclonal CD146 antibody demonstrated a weaker epithelial signal, which was significantly correlated with that of the polyclonal antibody, but revealed no prognostic value. However, the Western blot of the polyclonal antibody displayed a clearly reduced specificity. CONCLUSIONS: Evaluation of protein expression can be highly dependent on the primary antibody employed. A credible evaluation of antibody specificity is crucial to prove the validity of protein expression studies. The immunoreactivity of the polyclonal CD146 antibody (Abcam, ab28360) is prognostic of PSA-relapse in prostate cancer patients, although its immunoreactivity is possibly not restricted to CD146 associated epitopes.     

妊娠期高血压疾病患者CD146水平的检测及其与眼底视网膜血管病变的关系

目的 探讨不同类型妊娠期高血压疾病（HDIP）患者及正常妊娠产妇血清CD146水平与疾病严重程度及眼底视网膜血管病变分期的关系.方法 以2010年1月至2012年1月于首都医科大学附属北京同仁医院妇产科分娩的产妇113例为研究对象,包括HDIP患者73例和正常妊娠产妇40例.HDIP患者按照疾病严重程度分为3组：妊娠期高血压组（n=22）、轻度子痫前期组（n=23）、重度子痫前期组（n=28）;正常妊娠产妇为正常妊娠组（n=40）.酶联免疫吸附法（ELISA）检测各组产前和产后血清CD146水平,分析CD146水平与HDIP严重程度及眼底视网膜血管病变分期的关系.结果 产前、产后4组血清CD146水平差异均有统计学意义（均P＜0.05）,并且随着病情加重而升高.产前轻度子痫前期组、重度子痫前期组血清CD146水平高于正常妊娠组和妊娠期高血压组（均P＜0.05）.产后4组血清CD146水平均较产前下降（均P＜0.05）.所有研究对象按眼底视网膜病变不同分期分为4组,产前、产后4组血清CD146水平组间比较差异均有统计学意义（均P＜0.05）,并且CD146水平随眼底病变加重而升高.产前、产后眼底视网膜病变分期与HDIP严重程度之间均有显著的相关性（产前：rs=0.793,P＜0.05;产后：rs=0.631,P＜0.05）.结论 CD146水平可以作为反映HDIP及眼底病变严重程度的指标.     

慢性肾功能衰竭患者外周血细胞CD146的表达及意义

目的 探讨CD146在慢性肾功能衰竭(CRF)患者外周血细胞中的表达及意义.方法 收集本院2007年8月至2009年7月门诊及住院CRF患者51例为CRF组,33例正常人为健康对照组,流式细胞术检测两组外周血细胞CD146的表达并比较两组之间的差异,分析CRF患者外周血细胞CD146表达与血尿素氮(BUN)、血肌酐(Scr)的相关性.结果 与对照组比较,CRF组单核细胞、中性粒细胞CD146+细胞百分率及CD45-CD146+细胞数量均显著升高[(4.09±3.48)%比(0.20±0.10)%;(4.71±3.94)%比(0.79±0.14)%;(16.43±10.48)个/μl比(2.16±0.81)个/μl,均P<0.001].CRF患者外周血CD45-CD146+细胞数量与BUN、Scr均呈正相关(r=0.480、0.595,均P<0.01).结论 外周血单核细胞及中性粒细胞CD146的表达及升高与CRF的发生有关,CD45-CD146+细胞数量增加与CRF的发生及严重程度有关.     

Targeting the CD146/Galectin-9 axis protects the integrity of the blood–brain barrier in experimental cerebral malaria

Cerebral malaria (CM) is a life-threatening diffuse encephalopathy caused by Plasmodium falciparum, in which the destruction of the blood–brain barrier (BBB) is the main cause of death. However, increasing evidence has shown that antimalarial drugs, the current treatment for CM, do little to protect against CM-induced BBB damage. Therefore, a means to alleviate BBB dysfunction would be a promising adjuvant therapy for CM. The adhesion molecule CD146 has been reported to be expressed in both endothelial cells and proinflammatory immune cells and mediates neuroinflammation. Here, we demonstrate that CD146 expressed on BBB endothelial cells but not immune cells is a novel therapeutic target in a mouse model of experimental cerebral malaria (eCM). Endothelial CD146 is upregulated during eCM development and facilitates the sequestration of infected red blood cells (RBCs) and/or proinflammatory lymphocytes in CNS blood vessels, thereby promoting the disruption of BBB integrity. Mechanistic studies showed that the interaction of CD146 and Galectin-9 contributes to the aggregation of infected RBCs and lymphocytes. Deletion of endothelial CD146 or treatment with the anti-CD146 antibody AA98 prevents severe signs of eCM, such as limb paralysis, brain vascular leakage, and death. In addition, AA98 combined with the antiparasitic drug artemether improved the cognition and memory of mice with eCM. Taken together, our findings suggest that endothelial CD146 is a novel and promising target in combination with antiparasitic drugs for future CM therapies.     

CD146基因重组质粒的构建及蛋白表达

目的构建CD146原核、真核表达载体,证实融合蛋白在原核细胞的诱导表达以及在胃癌细胞内的表达和定位。方法提取人黑色素瘤细胞A875的总mRNA并进行反转。以反转录的cDNA为模板PCR扩增CD146全长编码基因,分别克隆至pCDNA3.1-myc/his以及pGEX-4T-3表达载体中。原核重组质粒鉴定后转入BL21细胞中并经了诱导表达及纯化,真核表达质粒转入胃癌MKN45细胞中,分别利用westernblot和激光共焦扫描显微技术检测了重组质粒的表达以及在胃癌细胞中的定位。结果 CD146全长基因序列克隆到原核、真核表达载体中,酶切鉴定片段为1930bp。原核诱导出了GST-CD146并进行了纯化,CD146在真核细胞中表达为113KD的糖蛋白,Westernblot检测到真核转染的myc/his-CD146表达,条带约为120KD,免疫荧光显示蛋白定位在胃癌MKN细胞膜。结论成功构建了CD146原核、真核表达载体,融合蛋白在胃癌MKN45细胞表达。     

Mouse CD146/MCAM is a marker of natural killer cell maturation

CD146/melanoma cell adhesion molecule is an adhesion molecule expressed by endothelial cells and by a small fraction of activated T and B lymphocytes in humans. In order to analyze the pattern of CD146 expression in mouse leukocytes at steady-state conditions, we generated a set of novel rat anti-mouse CD146 monoclonal antibodies. CD146 expression was undetectable on monocytes, dendritic cells, T cells or B cells, but was expressed on about 30% of neutrophils and 60% of NK cells. Within murine lymphocytes, CD146 was defined as a novel NK-specific surface molecule. An increased percentage of CD146+ cells was found in the most mature CD27(-)CD11b+ NK cell subpopulation, which also displays higher expression of Ly49C/I, Ly49D and KLRG1 and lower expression of NKG2A/C/E molecules. CD146+ NK cells were found to be less cytotoxic and produce less IFN-gamma than CD146(-) NK cells upon stimulation with target cells or activating antibodies. These findings define CD146 as a marker of mouse NK cell maturation that may be used as an alternative to the combined use of CD27 and CD11b staining to detect final stages of NK cell maturation.     

不同剂量黄芪注射液对人肾小管上皮细胞黏附分子CD146表达的影响

目的:探讨不同剂量黄芪注射液对人肾小管上皮细胞(HK2)黏附分子CD146表达的干预作用。方法:将传代HK2等分为5组,进行培养处理:正常对照组(LG组,只加5.5mmol/L D-葡萄糖),高糖组(HG组,加入25mmol/L D-葡萄糖),黄芪注射液干预组:在HG组中加入不同剂量黄芪注射液,加入2μg/ml者为低剂量组(AML组)、加入20μg/ml者为中剂量组(AMM组)、加入200μg/ml为高剂量组(AMH组)。观察37℃培养24h、48h、72h后,各组HK2形态变化、分别收集各组培养细胞,用Western Blotting检测其CD146蛋白表达。结果:HG组较LG组HK2变长,呈梭形,细胞间连接疏松,间隔增大;黄芪注射液干预各组,细胞形态均较HG组变小变圆,且连接逐渐紧密,AMH组呈堆积生长。HG组CD146表达均较LG组明显增加(P0.01),干预24h、48h、72h,AMM组、AMH组与HG组比较,CD146表达均明显减少(P0.05或P0.01),AML组干预培养72h,CD146的表达亦明显减少(P0.01)。结论:黄芪注射液改善高糖引起的HK2形态改变和降低其CD146表达,对防治DN有积极作用。