小胶质细胞在Cuprizone所致髓鞘脱失动物模型中的双重性作用

小胶质细胞(microglia)是中枢神经系统(CNS)中固有的免疫监视细胞,是CNS的重要组成部分,构成CNS的第一道免疫防线。小胶质细胞活化是许多CNS疾病的重要病理特征之一,其在多发性硬化(MS)的发病过程中扮演重要角色。MS是以炎症反应、髓鞘脱失和轴突损伤为主要特征的CNS疾病,MS患者及实验动物脑内均可见活化的小胶质细胞,其作用机制复杂,具有致炎和抗炎双重作用。一方面小胶质细胞可通过促进吞噬、轴突再生、释放神经营养因子等作用促进髓鞘的再生修复;另一方面小胶质细胞还可通过释放炎性因子、自由基、蛋白酶等对神经元和胶质细胞发挥毒性作用。Cuprizone(双环己酮草酰二腙)所致髓鞘脱失动物模型是研究髓鞘再生修复的理想模型,了解小胶质细胞的双重性对于理解MS的发生发展,寻找疾病治疗靶点具有十分重要的理论和实际意义。在此本文对小胶质细胞在cuprizone所致髓鞘脱失动物模型中的双重性作用做一综述。     

小胶质细胞在Cuprizone所致髓鞘脱失动物模型中的双重性

小胶质细胞(Microglia)是中枢神经系统(CNS)中固有的免疫监视细胞,是CNS的重要组成部分,构成CNS的第一道免疫防线。小胶质细胞活化是许多CNS疾病的重要病理特征之一, 其在多发性硬化(MS)的发病过程中扮演重要角色。MS是以炎症反应、髓鞘脱失和轴突损伤为主要特征的CNS疾病,MS患者及实验动物脑内均可见活化的小胶质细胞,其作用机制复杂,具有致炎和抗炎双重作用。一方面小胶质细胞可通过促进吞噬、轴突再生、释放神经营养因子等作用促进髓鞘的再生修复;另一方面小胶质细胞还可通过释放炎性因子、自由基、蛋白酶等对神经元和胶质细胞发挥毒性作用。Cuprizone(双环己酮草酰二腙)所致髓鞘脱失动物模型是研究髓鞘再生修复的理想模型,了解小胶质细胞的双重性对于理解MS的发生发展,寻找疾病治疗靶点具有十分重要的理论和实际意义。在此本文对小胶质细胞在Cuprizone所致髓鞘脱失动物模型的双重性做一综述。     

银杏内酯K通过抑制小胶质细胞介导的炎性反应保护髓鞘

目的:研究银杏内酯K(GK)能否通过抑制小胶质细胞介导的炎性反应保护髓鞘。方法:C57BL/6雄性小鼠随机分为正常(Normal)组、双环己酮草酰双腙(Cuprizone,CPZ)组和银杏叶内酯K组(GK组),通过免疫荧光染色评估各组髓鞘脱失程度和小胶质细胞的活化情况,Western blot和ELISA检测各组脑组织细胞因子诱导型一氧化氮合酶(iNOS)、精氨酸酶-1(Arg-1)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)、白介素-10(IL-10)和转化生长因子-β(TGF-β)的表达情况。结果:与正常组相比,CPZ成功诱导了小鼠胼胝体广泛的髓鞘脱失(P0.001),并引起了小胶质细胞的活化和炎性细胞因子iNOS、TNF-α及IL-6的升高(P0.001)。与CPZ组相比,GK组有效阻止了髓鞘脱失(P0.01),抑制了小胶质细胞的活化(P0.01)。同时GK组下调了促炎细胞因子iNOS、TNF-α和IL-6(P0.05),上调了抗炎细胞因子Arg-1、IL-10、和TGF-β(P0.05)。结论:GK通过抑制小胶质细胞介导的炎性反应保护髓鞘。     

CX3CL1/CX3CR1系统与神经系统疾病

趋化因子CX3CL1主要表达在神经元上，通过与表达在小胶质细胞上的趋化因子受体CX3CR1相互作用，调控神经元与小胶质细胞之间的信息传递。CX3CL1/CX3CR1系统成为近年来许多神经系统疾病研究的热点，目前有关该系统的研究报道尚存在争议。CX3CL1/CX3CR1系统可通过调节小胶质细胞活化或者促进小胶质细胞表型的转化等方式，在神经系统中发挥着神经保护或神经毒性的双重作用。本文就CX3CL1/CX3CR1系统在神经系统疾病中的作用及最新研究进展做一综述。     

抗人中性粒细胞单克隆抗体的筛选、鉴定和抑制调理吞噬活性的检测

用淋巴细胞分离液梯度离心法,自健康人血液中分离出中性粒细胞,作为颗粒抗原,免疫BALB/c小鼠,取脾细胞与SP2/0骨髓瘤细胞融合,获得分泌抗细胞膜单克隆抗体(单抗)杂交瘤细胞。用调理吞噬抑制试验,初步筛选出1株分泌抑制补体介导的吞噬功能的单抗杂交瘤细胞。单抗亚类检定结果为IgG_(2a)。     

β1-整合素与乳腺癌细胞凋亡关系的实验研究

目的探讨乳腺癌细胞凋亡与β1-整合素作用的关系.方法人乳腺癌细胞SK-BR-3在培养中以不同浓度的抗β1-整合素单克隆抗体的作用下,观察其对TNF-α诱导的凋亡并计算凋亡率.结果加入凋亡诱导剂后,凋亡细胞由正常生长的2.85±0.41增加到25.62±1.41(P＜0.01);而加入抗β1-整合素单克隆抗体的实验组凋亡细胞数明显减少,且随加入抗体浓度的增加而再度减少.结论TNF-α对SK-BR-3人乳腺癌细胞凋亡的诱导作用可能是由β1-整合素参与的介导作用实现的,且存在着一种在一定范围内的量、效依赖关系.     

豚鼠过敏性脑脊髓炎CNS中IL-1R的表达

目的 探索中枢神经系统脱髓鞘疾病进程中IL－1R的表达与神经胶质细胞活化的关系。方法 用牛脊髓髓鞘碱性蛋白加完全福氏佐剂对豚鼠进行免疫,诱发实验性过敏性脑脊髓炎（EAE）模型,采用免疫组化方法对EAE不同时期中枢神经系统中IL－1R的表达进行研究,同时采用抗蓖 凝集素RCA－1标记小胶质细胞、抗胶质纤维酸性蛋白标记星型胶质细胞。结果 在EAE症状出现之前3d至临床高峰期,小胶质细胞样细胞呈现IL－1R免疫反应阳性,表达IL－1R的星型胶质细胞样细胞呈活化状态,而恢复期则主要为星型胶质细胞样细胞呈现免疫反应阳性。结论 在EAE不同时期表达IL－1受体的细胞型不同与不同时期神经胶质细胞的活化类型有关,即与EAE早期小胶质细胞的活化和恢复期星型胶质细胞增生有关。     

不同状态下的小胶质细胞对脑缺血发病机制的作用

小胶质细胞作为中枢神经系统中最主要的免疫细胞,在缺血性损伤中涉及的炎症反应以及神经修复过程中起着重要的作用。生理情况下,小胶质细胞分泌营养因子对神经元提供营养支持,并参与神经环路的建立。脑缺血损伤时,小胶质细胞上的Toll样受体能识别并吞噬坏死的神经元,清除凋亡神经元;同时小胶质细胞也会释放出有害的细胞因子,加重神经元的损伤。脑缺血损伤发生时小胶质细胞会活化成两种状态：一种是促炎表型（M1型）：分泌白细胞介素-1(interleukin-1,IL-1),IL-6和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),加重炎症反应;一种是抗炎表型（M2型）：分泌IL-4,IL-10和TGF-β1,抑制炎症促进组织修复。脑缺血发生后,与小胶质细胞相关的通路例如NF-κB通路、TLRNF-κB通路、NLRP3炎性小体通路、Notch信号通路、p38MAPK信号通路等可能会被激活,因此药物通过不同的作用机制或者通路对小胶质细胞的活化产生影响,可能是一种新的治疗策略。     

大鼠视网膜光损伤模型中小胶质细胞的迁移及活化

 目的 探讨大鼠视网膜光损伤模型中小胶质细胞的迁移、活化及其与光感受器凋亡的关系。方法 将SD大鼠分为光照组（n=78）和正常对照组（n=15）,光照组大鼠在自制的光损伤箱中接受强度为2500 lux的宽谱蓝光照射24 h,建立光损伤模型。在光照结束后2 h、6 h、1天、3天、7天和14天时（每一时间点,n=13）,用原位末端标记法（TdT-mediated dUTP nick end labeling,TUNEL）染色观察两组大鼠视网膜细胞凋亡情况;用免疫荧光染色观察视网膜OX42(+)小胶质细胞的形态改变和迁移活动,并对视网膜外层的TUNEL(+)细胞和OX42(+)细胞分别计数并绘制时间-数量曲线;用透射电镜观察进入光感受器层的小胶质细胞的吞噬行为;用real-time PCR法定量分析光照后视网膜胶质源性神经毒性物质IL-1β mRNA的表达变化。结果 光照结束后2 h视网膜外核层（out nuclear layer,ONL）即可见TUNEL(+)细胞,1天后达高峰,3天后逐渐减少;光照结束后6 h视网膜ONL开始出现少量OX42(+)细胞,逐渐增多并于3天后达高峰,7天后渐消失,其形态转变为肥大细胞体的激活型。从时间-数量曲线可见OX42(+)细胞的迁移高峰落后并紧随凋亡高峰;强光照射上调了视网膜IL-1β mRNA的表达,其随时间变化的趋势同小胶质细胞的激活和迁移趋势基本一致;透射电镜显示进入ONL的小胶质细胞吞噬了光感受器的外节膜盘。结论 视网膜光损伤模型中光感受器的凋亡诱导小胶质细胞向ONL的迁移、活化及吞噬行为,并伴有视网膜IL-1β表达水平的升高,小胶质细胞的活化可能在加速光感受器变性过程中起重要作用。     

脊髓小胶质细胞在糖尿病神经痛形成中的作用

目的:观察大鼠糖尿病神经痛模型中脊髓小胶质细胞的活化状态和P物质含量的变化及腹腔注射米诺环素后的效应。方法:48只Wistar雄性大鼠,12只为正常对照组(C组);其余单次腹腔注射STZ制备糖尿病模型。自注射STZ前1 d起,连续29 d,糖尿病大鼠注入米诺环素40 mg/kg(M4组)、20 mg/kg(M2组)、等量PBS(D组)。在注射STZ前1 d、注射STZ后7 d、14 d、21 d、28 d(t1～5)测定机械性缩足反射阈值MWT。在t5时处死大鼠取脊髓,免疫组化测定P物质含量,RT-PCR测定小胶质细胞标志性蛋白CR3表达。结果:与C组相比,D组MWT下降,P物质含量降低,CR3 mRNA表达增强(P<0.05);与D组相比,M4组MWT升高,M4组和M2组P物质含量升高,CR3 mRNA表达减弱(P<0.05);与M2组相比,M4组MWT和P物质含量均升高(P<0.05),CR3 mRNA表达差异无统计学意义(P>0.05)。结论:糖尿病神经痛的形成依赖于脊髓内激活的小胶质细胞,P物质在该模型中含量减少,小胶质细胞活化被米诺环素抑制的同时,P物质含量增加并且神经痛得到了改善。     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

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.