外周血淋巴细胞穿孔素、颗粒酶B在大鼠肾移植急性排斥的表达及意义

目的:探讨大鼠同种异体肾移植外周血淋巴细胞(peripheralbloodlymphocyte,PBL)穿孔素、颗粒酶B的表达与急性排斥反应(acuterejection,AR)的关系。方法:实验组:异基因移植组(SD→Wister),对照组:同基因移植组(Wister→Wister)。采用逆转录聚合酶链反应(RT-PCR)检测移植肾和PBL穿孔素、颗粒酶B的表达,并以同基因肾移植为对照。结果:病理学检查结果显示实验组大鼠在术后第5、7、9、11天分别发生轻、中、重度排斥,对照组无明显排斥现象。RT-PCR结果显示急性排斥反应发生在肾移植术后3天,即急性排斥早期均表达穿孔素、颗粒酶B基因mRNA,4天后表达明显增强,并贯穿整个排斥反应全过程,而同基因对照组术后11天内均不表达两基因。外周血和肾组织中穿孔素、颗粒酶BmRNA的表达水平平均在急性排斥反应发生前3￣5天开始明显上升,其变化趋势早于血清肌酐。结论:穿孔素、颗粒酶B是一种有价值的AR标志物。定量RT-PCR测定外周血淋巴细胞中穿孔素、颗粒酶BmRNA表达可以较敏感预测肾移植急性排斥反应的发生,这一非侵入性检查方法可以为急性排斥反应的监测和诊断提供客观依据。     

外周血免疫效应分子基因表达与移植物急性排斥的关系

目的 探讨5种免疫效应分子基因的表达与移植肾急性排斥的关系.方法 采用定量逆转录PCR(RT-PCR)方法动态测定急性排斥反应(AR) 组(n=22)、阴性对照组(n=22)肾移植患者移植外周淋巴细胞(PBL)的颗粒酶B、穿孔素、Fas、FasL和TIA-1的含量,判断其与急性排斥的关系.结果 肾移植术后出现急性排斥的患者各效应分子表达均明显增强,在排异初期与后期有明显的下降(P<0.05);各效应分子升高时间比临床上出现AR的症状早约2d,随着AR的逆转,其表达也逐渐降至原有基础水平.AR组与对照组在多个基因表达的联合分析可不同程度地提高敏感性和特异性,3种基因同时上调尤其有意义.结论 定量逆转录PCR(RT-PCR)方法测定外周血免疫效应分子基因的表达,是一种无创、较敏感的早期诊断肾移植急性排斥的方法,并可预测抗排斥反应治疗效果.     

穿孔素和颗粒酶B蛋白表达对胰腺移植急性排斥反应早期诊断的研究

目的 研究移植胰腺局部穿孔素和颗粒酶B的蛋白表达对急性排斥反应的早期诊断和鉴别诊断的意义.方法 将40只四川当地杂种长白猪随机分为3组,即对照组(n=8)、移植组(n=16)、移植 免疫抑制剂治疗组(n=16).移植手术为同种异体经门静脉回流、肠内引流全胰十二指肠移植.免疫抑制剂治疗组在进行同种异体胰十二指肠移植术同时应用环孢A 骁悉 甲强龙三联免疫抑制治疗.分别于术前1 d和术后1、3、5、7 d检测受者的锁骨下静脉血胰岛素、胰高血糖素、血糖水平.术后1、3、5、7 d,开腹取胰腺组织进行常规病理学检查和免疫组化染色检测穿孔素和颗粒酶B的蛋白表达.结果 ①对照组、移植组以及移植 免疫抑制剂组之间受者的血糖、胰岛素、胰高血糖素水平变化有统计学意义(P＜0.05).②胰腺移植 免疫抑制剂组与单纯胰腺移植组相比胰腺组织中穿孔素和颗粒酶B的蛋白表达及穿孔素和颗粒酶B的阳性淋巴细胞计数的差异有统计学意义(P＜0.05).③免疫组化染色检测移植后胰腺组织中穿孔素和颗粒酶B蛋白表达,比常规病理学检查诊断排斥反应早2～4 d.结论 移植胰腺组织穿孔素和颗粒酶B局部蛋白表达变化早于常规病理学改变,结合常规病理学检查可以对胰腺急性排斥反应做出早期诊断和鉴别诊断.     

肾移植患者外周血淋巴细胞穿孔素的表达

目的：探讨外周血淋巴细胞(PBL)免疫标记物穿孔素在移植肾急性排斥反应(AR)诊断方面的价值。方法：选肾移植手术后近期稳定患者30例,AR18例,肾移植手术后长期稳定患者10例和健康对照组20例,应用聚蔗糖-泛影葡胺密度梯度离心法分离PBL,免疫细胞化学ABC法检测PBL中穿孔素表达。结果：AR患者PBL穿孔素表达阳性率明显高于肾功能近期与长期稳定组(60．95％ vs 16．66％)．并且穿孔素表达出现的时间可比AR的临床表现出现早2—3d,AR时应用甲基强的松龙冲击治疗可显著降低PBL穿孔素阳性细胞数。结论：穿孔素是一种有价值的AR标志物,对PBL中穿孔素表达的动态检查有助于早期诊断肾移植术后可能发生的AR。     

白细胞介素15和骨桥蛋白在大鼠肾移植急性排斥反应早期的表达

目的 研究白细胞介素(IL)15、骨桥蛋白、穿孔素及颗粒酶B在大鼠肾脏移植急性排斥反应(AR)早期的表达变化.方法 以BN大鼠和LEW大鼠分别作为供受体,建立大鼠肾移植动物模型.环孢素A处理组(CsA组):(BN→LEW,术后注射环孢素A);IL-15阻断组:(BN→LEW,术后注射IL-15抗体);AR组:(BN→LEW,术后注射生理盐水);对照组:(LEW→LEW,术后注射生理盐水).对照组6只大鼠,其余组10只大鼠,于移植后1、3、5、7 d取外周血,逆转录一聚合酶链反应(RT-PCR)动态检测骨桥蛋白、IL-15、穿孔素及颗粒酶B mRNA的表达,并作移植肾病理分析.结果 AR组、CsA组及IL-15阻断组肾移植术后IL-15、骨桥蛋白mRNA表达逐渐上调,第5天均达到高峰;对照组均处于低水平.术后第5天,CsA组和IL-15阻断组IL-15 mRNA分别为9685±1440、4346±741均低于AR组(17 022±2153,P<0.01),IL-15阻断组明显低于CsA组(P<0.01);CsA组和IL-15阻断组骨桥蛋白mRNA分别为13 226±1565、19 112±2908均低于AR组(24 663±2449,P<0.01),IL-15阻断组明显高于CsA组(P<0.01).术后第5天IL-15阻断组和CsA组穿孔素和颗粒酶B mRNA表达明显低于AR组(P<0.01).移植肾病理组织检查显示,AR组呈现重度急性排斥反应,IL-15阻断组排斥反应征象轻微.结论 大鼠肾移植AR发生早期骨桥蛋白、IL-15、穿孔素和颗粒酶B mRNA在外周血中均高水平表达,通过阻断IL-15/IL-15受体途径可明显下调颗粒酶B和穿孔素mRNA的表达.     

外周血穿孔素和颗粒酶B mRNA检测对胰腺移植急性排斥反应早期诊断的价值

目的 研究胰腺移植术后外周血穿孔素和颗粒酶B的基因表达对急性排斥反应早期诊断的价值.方法 实验动物为杂种长白猪,分为3组(n=8):对照组,移植组.免疫抑制剂治疗组.移植手术为同种异体门静脉回流、肠内引流全胰十二指肠移植.免疫抑制剂治疗组在行同种异体胰十二指肠移植的同时应用环孢A 骁悉 甲强龙三联免疫抑制治疗.分别于术前1 d和术后1、3、5、7 d采集受者锁骨下静脉血,抽提总RNA,RT.PCR方法检测静脉血淋巴细胞中穿孔素和颗粒酶B的mRNA表达,检测受者血糖、胰岛素、胰高血糖素水平.在术后1、3、5、7d,开腹取胰腺组织进行常规病理学检查,与血液中穿孔素和颗粒酶B的mRNA表达水平进行比较分析.结果 ①对照组、移植组、免疫抑制剂组之间受体的血糖、胰岛素、胰高血糖素水平变化差异无统计学意义.②与移植组比较,免疫抑制剂组静脉血淋巴细胞中穿孔索和颗粒酶B的mRNA表达水平下降(P<0.05).③胰腺移植术后静脉血穿孔素、颗粒酶B mRNA表达变化比急性排斥反应病理学改变早2～d.结论 监测外周血淋巴细胞中穿孔素和颗粒酶B mRNA表达的变化,可以对胰腺急性排斥反应做出早期诊断.     

外周血CTL活化基因在大鼠肾移植急性排斥的表达及意义

目的：探讨大鼠同种异体肾移植外周血淋巴细胞（peripheral blood lymphocyte，PBL）穿孔素、颗粒酶B和FasL mRNA表达与急性排斥反应（acute reiection，AR）的关系。方法：采用TdT介导的脱氧核苷酸原位末端标记法（TUNEL）检测移植肾脏中的凋亡细胞。另外同时检测血清肌酐水平。采用逆转录聚合酶链反应（RT-PCR）检测移植肾和PBL中穿孔素、颗粒酶B和FasLmRNA表达。并以同基因肾移植为对照。结果：病理学检查结果显示实验组大鼠在术后第5、7、9～11天分别发生轻、中、重度排斥。对照组无明显排斥现象。实验组于手术后第5、7、9～11天细胞凋亡数明显高于对照组（P〈0．01）：肾小管上皮细胞凋亡程度与血清肌酐水平、病理学检查结果一致。RT-PCR结果显示实验组于术后3～5天起，肾组织和PBL中穿孔素、颗粒酶B和FasLmRNA表达水平即显著升高，于术后7-9天达到高峰。与对照组相比差异有显著性（P〈0．01）。外周血和肾组织中FasL、穿孔素和颗粒酶BmRNA的表达水平平均在急性排斥反应发生前3～5天开始明显上升，其变化趋势早于血清肌酐。结论：定量RT-PCR测定外周血淋巴细胞中穿孔素、颗粒酶B和FasLmRNA表达可以较敏感预测肾移植急性排斥反应的发生，具有临床诊断参考价值。     

肺癌患者血清对植物血凝素活化的淋巴细胞表达穿孔素和颗粒酶B的抑制作用

目的探讨肺癌患者血清对植物血凝素(PHA)活化的淋巴细胞表达穿孔素和颗粒酶B的影响。方法收集肺癌患者外周静脉血,分离血清和淋巴细胞,用肺癌患者血清作用于PHA活化的淋巴细胞,采用免疫细胞化学染色及Western blot技术测定淋巴细胞穿孔素和颗粒酶B的表达,以健康血清为对照。结果免疫细胞化学染色显示,在肺癌患者血清的作用下,PHA活化的淋巴细胞穿孔素和颗粒酶B的表达明显低于对照组;Western blot技术也显示,在肺癌患者血清的作用下,PHA活化的淋巴细胞穿孔素和颗粒酶B的表达明显低于对照组,差异有统计学意义(分别为t=11.105,P=0.000;t=13.885,P=0.000)。结论肺癌患者血清对PHA活化的淋巴细胞表达穿孔素和颗粒酶B具有抑制作用。     

肝移植术后趋化因子Mig、IP10、ITAC的变化对早期诊断急性排斥反应的意义

目的:观察趋化因子Mig、IP10、ITAC在肝移植术后的变化,探讨其对肝移植术后急性排斥早期诊断的意义.方法:2005年4～9月30例肝移植患者,根据临床表现及病理学检查分为急性排斥组(AR,n=9)(排除3例术后感染患者),非急性排斥组(NAR,n=18).检测患者术前1 d及术后1、3、5、7 d血清中趋化因子Mig、IP10、ITAC的表达,并与同期住院的肝硬化或肝癌患者(肝癌肝硬化组,n=16)以及健康体检者(正常对照组,n=16)进行比较.AR组患者分别于确诊当天以及经激素冲击治疗后3、7 d检测血清中3种趋化因子的表达,并分析确诊当天3种趋化因子表达与肝脏穿刺活检Banff排斥活动指数(RAI)的相关性.结果:肝移植术前1 d AR组、NAR组患者血清Mig、IP10、ITAC表达与肝癌肝硬化组无明显差异,但高于正常对照组(P<0.01).肝移植术后3 d,AR和NAR组Mig、IP10、ITAC的表达水平均有上升,高于术前1 d的表达(P<0.05).AR组9例患者分别在术后11、12和14 d确诊AR发生,术后各时间点患者血清Mig、IP10、ITAC表达明显高于NAR组(P<0.01);确诊AR当天Mig、IP10、ITAC的表达与RAI呈正相关(r=0.88、0.94、0.80).与确诊AR当天相比,经冲击治疗逆转后,3种趋化因子的表达也相应下降(P<0.01).结论:肝移植术后血清中趋化因子Mig、IP10、ITAC的表达可作为早期诊断急性排斥反应辅助特异、敏感的指标.     

类风湿性关节炎患者CTL和NK细胞颗粒酶素B和穿孔素的表达及临床意义

目的:检测类风湿性关节炎(rheumatoid arthritis,RA)患者外周血T杀伤细胞(cytotoxic T lymphocyte, CTL)和自然杀伤细胞(NK)中颗粒酶素B和穿孔素的表达,探讨其可能的临床意义.方法:取25例健康者、43例RA患者(31例活动期、12例稳定期;初发16例、复发27例)外周血及其中15例患者(初发8例、复发7例)关节液标本.采用三色流式细胞术胞内检测外周血及关节液标本中CTL、NK细胞颗粒酶素B和穿孔素的表达.结果:RA患者关节滑膜液中CTL和NK细胞颗粒酶素B及穿孔素的表达显著高于RA患者健康者外周血(P＜0.01或P＜0.05).活动期RA患者外周血CTL和NK细胞表达颗粒酶素B、穿孔素明显高于稳定期RA患者(P＜0.01或P＜0.05).初发和复发患者关节液及外周血中二者表达无显著差异.结论:RA患者外周血及关节液中CTL和NK细胞颗粒酶素B及穿孔素的表达显著高于正常人外周血,对其水平的检测利于判断疾病活动程度,但对判断疾病初发、复发的意义不大.     

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.     

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.     

Electro-Acupuncture Combined with the Trigger Point Needle-Embedding for Treatment of Primary Trigeminal Neuralgia in 31 Cases

In recent years, the author of this essay has applied electro-acupuncture combined with the trigger point needle-embedding for treatment of primary trigeminal neuralgia in 31 cases, yielding satis- factory results as reported in the following.     

Investigation of Matrix Metalloproteinase -1, 2 and tissue inhibitor of matrix metalloproteinases-1 in Endometriosis

Objective： To explore the role of matrix metalloproteinase-1,2 （MMP-1, MMP-2） and tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） in endometriosis. Methods： The eutopic and ectopic endometria from 40 subjects suffering from endometriosis and regular.endometria from 40 subjects （excluding endometriosis） were collected and examined by in situ hybridization technology and western blot assay. Results： Both expressions of MMP-1 and -2 were stronger in ectopic endometrium and eutopic endometrium than in normal endometrium. On the contrary, the expression of TIMP-1 in ectopic endometrium and eutopic endometrium was lower. The differences were significant （P 〈 0.01 ）. Moreover, there was no relationship among the expressions of MMP-1, 2 and TIMP-1 in ectopic endometrium. Conclusion： The expressions of MMP-1, 2 and TIMP-1 lose balance and lack of periodic changes in ectopic endometrium , which explains the biological invasive behavior of endometriosis. It was suggested-that regulating the balance between the MMPs and TIMP-1 should be an ideal therapeutic target to endometriosis.     

Shi Da-zhuo (史大卓) —An outstanding cardiologist of integrative medicine

Prof. SHI Da-zhuo, Ph.D., male, was born on March 20, 1960. Prof. SHI entered the Ph.D. program in 1990 at the China Academy of Chinese Medical Sciences under the supervision of Prof. CHEN Ke-ji, majoring in the treatment of cardiovascular diseases. After receiving his Ph.D. degree in 1993, Prof. SHI started working at the Cardiovascular Center in Xiyuan Hospital affiliated to China Academy of Chinese Medical sciences.     

Persist in and Carry Forward the Integrative Medical Research——Speech at the 6th National General Congress of Chinese Association of Integrative Medicine by Academician CHEN Zhu,Minister of Ministry of Health

The 6th National General Congress of Chinese Association of Integrative Medicine （CALM） was convened at 19-20, April 2008 in Beijing. Academician CHEN Zhu, the minister of Ministry of Health indicated at the congress that the integration of Chinese and Western medicine is very well in keeping with the situation of our country and the general rule of development in medical science; and as a good integration of Chinese medicine and Western medicine, it is mutually beneficial and advantageous to both of them. Seeing the creativity shown in integrative medical investigation in theoretic and methodological sides, we should and must persist in and develop it.