应用SELDI-TOF-MS技术筛选2型糖尿病肾病血清差异表达蛋白

目的 采用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术建立2型糖尿病肾病血清蛋白指纹图谱,筛选其差异表达蛋白.方法 收集2型糖尿病肾病(T2DN)患者31例、2型糖尿病(T2DN)不伴肾病患者32例和正常对照血清30例,采用SELDI-TOF-MS技术建立蛋白指纹图谱,应用BioMarker Wizard 3.1软件对实验数据进行分析.结果 糖尿病肾病组与糖尿病不伴肾病组相比较,有14个差异表达蛋白峰(P＜0.05),其中糖尿病肾病组有2个蛋白峰高表达,12个蛋白峰低表达;糖尿病肾病组与正常对照相比较,有25个差异表达蛋白峰(P＜0.05),其中糖尿病肾病组有13个蛋白峰高表达,12个蛋白峰低表达.结论 糖尿病肾病具有多种差异表达蛋白,这些蛋白可能为其诊断的潜在标志物.     

阳和汤对急性乳腺炎大鼠蛋白质组的影响

目的探究阳和汤对急性乳腺炎大鼠蛋白质组的影响,从而探索阳和汤治疗急性乳腺炎可能的途径。方法采用双向凝胶电泳(2-DE)对实验对照组、模型组、青霉素组和阳和汤组大鼠血清进行蛋白分离,经过图像分析识别差异表达蛋白质,由基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)得到差异表达蛋白的肽指纹图,用蛋白质数据库(SwissProt)搜索匹配的蛋白质。结果所获2-DE图谱分离效果较好,质谱分析共鉴定了12个差异蛋白质点,分别属于炎症相关蛋白、物质代谢相关蛋白和肿瘤相关蛋白,青霉素组的差异蛋白有三个上调,九个下调,阳和汤组的差异蛋白全部下调。结论阳和汤治疗急性乳腺炎可能与下调炎症相关蛋白、物质代谢相关蛋白和肿瘤相关蛋白有关。     

糖尿病肾病大鼠CTGF异常表达及药物干预研究

目的 探讨缬沙坦和雷公藤多甙对糖尿病肾病大鼠结缔组织生长因子(CTGF)异常表达的干预作用.方法 60只雄性SD大鼠随机分为正常对照组、糖尿病肾病组(DN组)、糖尿病肾病缬沙坦干预组(VAL组)和雷公藤多甙干预组(TG组).DN组、VAL组和TG组大鼠经腹腔注射链脲佐菌素(STZ)建立糖尿病肾病模型.于实验2、4、8、12周后分别处死大鼠,测定大鼠体质量、血糖、肾功能、尿白蛋白排泄率(UAER);RT-PCR检测肾脏CTGF mRNA表达.结果 与正常对照组相比,DN组大鼠血糖、血尿素氮、血清肌酐和UAER显著增高(P＜0.05或P＜0.01),肾脏CTGFmRNA表达增加(P＜0.01).VAL组和TG组除血糖外的各项检测指标均较DN组明显改善(P＜0.05或P＜0.01),而VAL组与TG组组间比较无显著性差异.结论 缬沙坦和雷公藤多甙对DN模型的干预效果相似,两者对糖尿病大鼠肾脏保护作用可能与减少肾脏CTGF表达相关.     

糖尿病肾病大鼠CTGF异常表达及药物干预研究

目的 探讨缬沙坦和雷公藤多甙对糖尿病肾病大鼠结缔组织生长因子(CTGF)异常表达的干预作用.方法 60只雄性SD大鼠随机分为正常对照组、糖尿病肾病组(DN组)、糖尿病肾病缬沙坦干预组(VAL组)和雷公藤多甙干预组(TG组).DN组、VAL组和TG组大鼠经腹腔注射链脲佐菌素(STZ)建立糖尿病肾病模型.于实验2、4、8、12周后分别处死大鼠,测定大鼠体质量、血糖、肾功能、尿白蛋白排泄率(UAER);RT-PCR检测肾脏CTGF mRNA表达.结果 与正常对照组相比,DN组大鼠血糖、血尿素氮、血清肌酐和UAER显著增高(P＜0.05或P＜0.01),肾脏CTGFmRNA表达增加(P＜0.01).VAL组和TG组除血糖外的各项检测指标均较DN组明显改善(P＜0.05或P＜0.01),而VAL组与TG组组间比较无显著性差异.结论 缬沙坦和雷公藤多甙对DN模型的干预效果相似,两者对糖尿病大鼠肾脏保护作用可能与减少肾脏CTGF表达相关.     

双向电泳结合质谱法分析双相情感障碍患者血浆差异表达蛋白

目的 应用比较蛋白质组学分析静息相双相情感障碍(bipolar disorder,BD)患者和健康对照血浆的差异表达蛋白质,以期找到与情感发作无关但和疾病发生相关的蛋白质.方法 收集BD静息相患者(n=12)和健康对照者(n=20)空腹血浆,MARS柱去除7种高丰度蛋白质,运用2-DE分离血浆蛋白,PD-Quest分析后MALDI-TOF/TOF鉴定差异表达的蛋白点,MetaCore软件进行生物信息学分析,Western blot验证部分差异蛋白结果.结果 经2-DE分析在静息相BD患者血浆中找到45个差异蛋白点,质谱鉴定出27个非冗余蛋白质,生物信息学分析显示差异蛋白主要与免疫调节相关,Apo A1和A2M经Western blot验证差异与2-DE结果一致且有统计学意义(P＜0.05).结论 与健康对照相比,BD静息相患者存在蛋白质谱的异常表达.差异表达蛋白提示BD静息相患者存在免疫功能异常及脂质代谢紊乱.     

BMP-2在糖尿病肾病大鼠肾内小动脉中表达的意义

目的 观察骨形态蛋白2(BMP-2)在糖尿病肾病(DN)大鼠肾实质内小动脉上的表达及其变化规律,探讨其对糖尿病肾病血管病变的影响.方法 设对照组(N组)和糖尿病肾病组(DN组),建立链脲佐菌素(STZ)诱导的DN大鼠模型.茜素红染色观察肾实质内小动脉周围钙盐沉积,并分别采用免疫组化及mRNA 原位杂交法检测BMP-2在肾实质内小动脉上的蛋白表达及基因定位.结果 (1)茜素红染色DN 组第4～12周时偶见肾内小动脉少许点状红色钙盐沉积,第24 周时可见肾实质内小动脉及周围组织有较多红色点片状沉积,N 组茜素红染色为阴性.(2)DN 组肾实质内小动脉BMP-2蛋白及mRNA 在4周时已有表达,随时间延长表达逐渐增强,各时间点均明显高于N组,N组各时间点表达差异无统计学意义.结论 (1)DN大鼠肾内小动脉钙化前即已有BMP-2的表达;(2)BMP-2可能是糖尿病血管钙化的重要转录因子及早期生物学标志之一; (3)肾内小动脉上BMP-2的表达变化可能参与DN进展.     

实时荧光定量PCR法检测糖尿病肾病大鼠肾脏中BMP-2与MGP

目的:检测糖尿病肾病(DN)大鼠肾脏中骨形成蛋白2(BMP-2)和基质Gla蛋白(MGP)的基因表达量的变化规律,探讨其对糖尿病肾病的影响.方法:设对照组(N组)及糖尿病肾病组(DN组,STZ诱导的DN大鼠模型);采用实时荧光定量PCR(Real-Time PCR)方法检测4、12、24周DN大鼠及正常大鼠肾脏中BMP...     

早期糖尿病肾病肾阳虚证的血浆蛋白质组学分析

目的寻找糖尿病肾病肾阳虚证敏感的血浆分子标志物。方法对4例早期糖尿病肾病肾阳虚证患者的血浆和4例正常人血浆进行荧光差异双向电泳（2－D DIGE）分析,选择表达差异大于1．5倍的蛋白斑点进行质谱分析。结果成功建立糖尿病肾病肾阳虚证血浆和正常血浆的胶内差异双向凝胶电泳图谱,通过分析共鉴定出9种差异蛋白质,包括补体C3、补体C4、载脂蛋白E、泛素化因子等。结论荧光差异双向电泳能够客观地显示糖尿病肾病肾阳虚证血浆与正常血浆之间的蛋白质表达差异,经鉴定的9种蛋白有可能为糖尿病肾病的早期诊断以及中医证型研究提供潜在的血浆分子标志物。     

吗啡成瘾和正常大鼠前额叶皮质差异表达蛋白的比较研究

目的：比较吗啡成瘾与正常大鼠前额叶皮质（prefrontal cortex,PFC）蛋白质双向电泳图谱,寻找和鉴定吗啡成瘾大鼠PFC中的差异表达蛋白质。方法：以固相pH梯度等电聚焦为第一向和垂直SDS-PAGE为第二向,分别对正常对照大鼠和吗啡成瘾大鼠的PFC蛋白质样品进行二维分离,2-DE图谱经ImageMaster 2D Platinum v5.0软件分析,选取4个差异蛋白点用基质辅助激光解吸附离子化飞行时间质谱（MALDI-TOF-MS）进行鉴定。结果：通过对2-DE图谱蛋白斑点的匹配及对比分析,与吗啡成瘾相关的差异表达蛋白斑点为87个;经质谱鉴定出2个有意义的差异表达的蛋白斑点：snap25亚型β-snap25突触相关蛋白25、核不均一核糖核蛋白U。结论：吗啡成瘾组与正常对照组大鼠PFC蛋白质组存在差异;初步鉴定出了2种大鼠前额叶皮质中与吗啡成瘾相关的差异蛋白,其表达的变化可能通过多种途径影响PFC神经元功能,为研究阿片类物质依赖作用机制提出了新的思路和方向。     

西维来司钠对脑缺血/再灌注大鼠全脑蛋白质组的影响

目的:研究西维来司钠(Sivelestat sodium hydrate,ONO-5046)对脑缺血/再灌注(Cerebral ischemia/reperfusion,CI/R)大鼠全脑蛋白质表达的影响.方法:大鼠随机分为假手术组、CUR组和CI/R+ONO-5046组(6 mg/kg).采用四血管阻断法制备CUR模型,缺血15 min,再灌注24 h.提取脑组织总蛋白进行双向电泳,对差异蛋白点用HPLC-Chip-MS/MS纳流液质联用技术进行序列分析,经Spectrum Mill搜索NCBInr数据库后鉴定蛋白质.结果:成功获得了分辨率和重复性好的大鼠脑组织蛋白2-DE图谱,经质谱鉴定最终获得了巨噬细胞炎性蛋白1-beta、白细胞弹性蛋白酶A、β2-巨球蛋白、谷胱甘肽S-转移酶omega-1、谷氨酸脱氢酶-1、细胞色素C还原酶铁硫亚单位、硫氧还蛋白过氧化物酶1等19个差异蛋白点相关信息.结论:大鼠脑缺血后脑组织蛋白质表达存在明显差异,西维来司钠可通过调节有关蛋白质的表达,参与大鼠脑缺血神经元保护作用.     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.