免疫细胞在小鼠→大鼠心脏移植排斥反应中的作用

目的 :探讨T细胞、巨噬细胞 (MΦ)和自然杀伤细胞 (NK)细胞在小鼠→大鼠心脏移植反应中的作用 .方法 :动物随机分 4组 :对照组、环孢素A(CsA)组、环磷酰胺 (CyP)组和CsA +CyP组 .排斥后取移植心脏作病理学检查 .用免疫组织化学法检测CD4 ,CD8,CD5 7和CD6 8在移植物中的沉积 .结果 :单用CsA (2 .6± 1.5 )d或CyP(3.2± 1.6 )d均不能显著延长异种移植物的存活时间 ,联用CsA和CyP使移植心脏存活 (4 .7± 1.4 )d (P <0 .0 5 ) .病理检查发现被排斥心脏中大量细胞浸润 ,所有被排斥心脏中均有大量NK细胞和MΦ浸润 ,但未见CD4 +和CD8+T细胞浸润 .结论 :NK和MΦ在小鼠→大鼠心脏移植反应中起重要作用 ,而与T细胞无明显相关 .     

耐受T细胞抑制NK细胞和巨噬细胞对大鼠皮肤移植物的排斥

目的 了解NK细胞和巨噬细胞在T细胞缺乏或T细胞成功重建的情况下对异种移植物排斥的作用。方法在无胸腺裸小鼠、通过混合胸腺移植重建T细胞后裸小鼠和正常免疫健全小鼠分别进行F344大鼠皮肤移植,观察异种皮肤移植物的存活情况,并利用流式细胞仪检测各组受体动物外周血中CD4+和CD8+T细胞的百分率,利用ConA增殖实验和混合淋巴细胞培养检测其体外功能状态。结果 在没有清除受体体内NK细胞和巨噬细胞的情况下,正常无胸腺BALB/c裸小鼠上移植的F344大鼠皮肤移植物发生了排斥,平均存活时间为(20.50±2.38)d;而在裸小鼠左肾包膜下移植的F344胎大鼠胸腺组织,却能够正常发育,并有效重建了受体的T细胞免疫,而且诱导了F344大鼠皮肤移植物的长期存活。结论NK细胞和巨噬细胞在异种移植物的排斥中起十分重要的作用,不同组织对NK细胞和巨噬细胞细胞毒性的易感性可能存在一定的差异。对异种供体抗原已产生耐受的T细胞可抑制受体体内NK细胞和巨噬细胞对异种移植物的排斥。     

FTY720与ICAM-1单抗对小鼠-大鼠异种心脏移植抗排斥的作用

目的:研究FTY720与ICAM-1单抗(mAb)对小鼠-大鼠异种心脏移植的抗排斥作用.方法:采用小鼠-大鼠腹部心脏移植,观察移植心存活时间、病理检查、CD4和CD8细胞的浸润程度及检测血中IL-2,IFN-γ,IL-4以及IgM抗体水平.结果:对照组移植心平均存活时间为(2.75±0.43)d.单用ICAM-1 mAb组移植心存活时间无明显延长,单用FTY720大剂量组移植心平均存活(4.25±0.71)d(P＜0.01):FTY720与ICAM-1 mAb大剂量联合用药组移植心存活时间平均(10.25±2.1 2)d(P＜0.01).联合用药组的病理反应、T细胞浸润及IL-2,IFN-γ,IgM水平明显减轻或降低;IgM水平与移植物存活时间呈负相关(R=-0.754,P＜0.01).结论:FTY720与ICAM-1 mAb单一用药对异种移植的抗排斥作用不明显,联合用药可有效延长异种心脏移植的存活时间并能抑制异种抗体的产生.     

供体血管内皮细胞胸腺修饰对猪-猴心脏移植的影响

目的　探讨胸腺修饰对大动物异种心脏移植的影响 ,建立猪—猴异种心脏移植模型。方法　受体猕猴随机分成 4组 :对照组、照射组、照射 +胸注 16 4 0组和照射 +胸注供体血管内皮细胞 (VECs)组 ,对受体进行不同预处理后 ,作猪—猴腹腔异位心脏移植。同时在移植前作供体VECs溶解率和供体VECs与受体淋巴细胞 (猴T细胞 )的混合细胞反应检测。结果　照射 +胸注猪VECs组平均供心存活时间 (MeanSurvivalTime,MST)显著延长(12 4 .2± 4 .2h) ,超过照射组 (6 5 .2± 6 .6h)、照射 +胸注 16 4 0组 (6 3.0± 4 .0h)和对照组 (36 .2± 5 .9h) (P <0 .0 5 )。同时 ,该组供体VECs溶解率显著下降 (P <0 .0 5 ) ,受体T细胞增殖显著受抑制 (P <0 .0 5 )。结论　供体VECs(猪VECs)胸腺修饰可通过抑制受体 (猴 )T细胞的识别与增殖 ,减轻供心VECs激活 ,最终缓解延迟性异种排斥反应(DelayedXenograftRejection ,DXR) ,从而使供心存活时间显著延长 ,但不能完全克服DXR。     

协调性异种心脏移植排斥反应病理变化的动态研究

目的 建立小鼠→大鼠异种心脏移植延迟性异种移植排斥反应(delayed xenograft rejection, DXR)动物模型,动态观察移植心脏组织病理和免疫病理变化,探讨DXR发生机制.方法 袖套法建立小鼠→大鼠颈部异位异种心脏移植模型,设立未移植小鼠心脏为对照组(n=4),余组分别在移植术后3、8、16、24 h(每时相点n=4)供心仍跳动时取心,另一组(n=16)在供心停跳时取心并以此决定移植心脏存活时间.各组标本作HE染色和SABC法检测补体C3、IgM、IgG、E-selectin和巨噬细胞标记物CD68表达,结果行半定量评价.结果 移植心脏HE染色表现为血管内皮损伤、血栓形成、心肌实质损伤、间质出血、炎症细胞浸润等排斥反应随移植术后时间逐渐加重趋势,完全排斥时部分心肌凝固性坏死及溶解、广泛间质出血、血管结构崩解及血管内血栓形成、严重炎症细胞浸润.移植术后供心平均存活时间(49.3±16.2) h.移植心脏免疫组化检测示:移植后任何时段均无C3沉积;从移植后3 h开始即有显著IgM沉积,且IgG沉积也逐渐增加;移植后3 h即有E-selectin表达,且表达逐渐增强,排斥时达高峰;从移植后3 h 开始在浸润的炎症细胞中有逐渐增加的CD68阳性细胞.结论 小鼠→大鼠异种心脏移植可作为DXR研究动物模型.内皮细胞激活、IgM和IgG抗体、巨噬细胞浸润参与DXR发生,但其发生不依赖于补体参与.     

中华眼镜蛇毒因子清除补体建立异种心脏移植急性血管性排斥反应模型

目的　采用纯化中华眼镜蛇毒因子 (CVF)清除补体 ,建立豚鼠 -大鼠非协调性异种心脏急性血管性排斥反应 (AVR)模型。　方法　供者豚鼠和受者 SD大鼠各 38只 ,分为 3组 : 组 (超急排组 ,n=2 0 ) ,受者不进行任何预处理 ; 组 (小剂量 CVF组 ,n=1 0 ) ,受者移植前 1 d经腹腔注射 CVF2 0 0 μg/ kg,移植后每日注射 1 0 0 μg/kg直至供心停搏 ; 组 (大剂量 CVF组 ,n=8) ,受者移植前 1 d经腹腔注射 CVF4 0 0 μg/ kg,移植后每日注射 2 0 0μg/ kg直至供心停搏。观察供心存活时间 ,供心停跳后行病理组织学检查。　结果　 组和 组供心平均存活时间分别为 (4 1± 2 ) h和 (4 0± 3) h明显长于 组 [(1 7± 2 ) min,P<0 .0 1 ], 组与 组间差异无显著性 (P>0 .0 5 )。病理检查 : 组呈超急性排斥反应 (HAR)改变 ,而 、 组呈 AVR改变 ; 、 组移植物炎症细胞浸润数明显多于 组(P<0 .0 1 ) ,移植物补体 C3沉积明显少于 组 (P<0 .0 1 )。　结论　适当剂量纯化 CVF具有良好的清除大鼠补体C3作用 ,克服豚鼠—大鼠非协调性异种心脏移植物 HAR的发生 ,延长其存活时间 ,建立以炎症细胞浸润为主的异种移植 AVR模型     

小鼠腹部同种异体心脏移植模型的建立及改进

目的 :建立并改进小鼠腹部同种异体心脏移植模型。 方法 :在放大 2 6倍视野下用 10 0线将小鼠供体心脏的升主动脉和肺动脉分别与受体腹主动脉和下腔静脉行端侧吻合 ,术后观察移植心存活时间并做病理检查。结果 :手术成功率达 90 % ,平均取心时间和血管吻合时间分别为 (5 .9± 0 .97)min和 (38.5± 5 .5 )min ;Balb/c小鼠 昆明小鼠移植心平均存活时间为 (6 .2± 1.0 3)d ,昆明小鼠 昆明小鼠移植心存活时间超过 2 0d。 结论 :小鼠心脏移植模型与大鼠模型相比 ,技术操作难度大 ,术者需更长时间的训练 ,可作为大鼠心脏移植模型的补充 ,是利用纯系鼠进行移植研究的廉价选择     

茯苓醇提取物抗心脏移植急性排斥反应的实验研究

目的　研究茯苓醇提取物对心脏移植急性排斥反应的抑制作用。方法　以Wistar大鼠为供者、SD大鼠为受者 ,建立异位 (腹腔 )心脏移植模型 ,移植前按组分别以橄榄油、茯苓醇提取物及环孢素A灌胃 ,并设SD大鼠空白对照组。术后观察移植心的存活时间 ,测定各组术后第 7d外周血中白细胞介素 2 (IL 2 )和γ干扰素 (IFN γ)含量以及CD3+ 、CD4 + 、CD8+ 细胞和CD4 + 细胞与CD8+ 细胞的比值 (CD4 + CD8+ ) ,并观察移植心的病理变化。结果　按受茯苓醇提取物 2 5mg·kg- 1 ·d- 1 或 5 0mg·kg- 1 ·d- 1 灌胃的大鼠 ,移植心存活时间显著延…     

IL-17参与小鼠心脏移植排斥反应的实验研究

目的 研究Th17细胞相关因子白细胞介素17(IL-17)在小鼠心脏移植排斥反应中的表达及意义.方法 建立小鼠颈部异位心脏移植模型,实验动物随机分为同系移植组和急性排斥反应组.观察2组供心存活时间,应用RT-PCR检测移植心脏IL-17、核孤独受体γt(RORγt)、干扰素-γ(IFN-γ)mRNA在移植术后1、2、3、4、5、6、7、8 d的动态表达水平.免疫荧光法观察移植术后第4天移植心脏内CD4+、CD8+T细胞中IL-17的表达情况.结果 RT-PCR检测显示,在急性排斥反应组中,IL-17和RORγt mRNA的表达都早于IFN-γ,在移植术后第2天即可于移植心脏中检测到,其表达量在术后第4天均达到高峰,随后逐渐下降,并分别在术后第6天和第7天无表达.免疫荧光结果显示,急性排斥组移植心脏内有大量CD4+、CD8+T细胞浸润,细胞因子IL-17主要由CD4+T细胞分泌.结论 Th17细胞在移植排斥反应的发生发展中可能起着非常重要的作用,对移植脏器中细胞因子IL-17的检测可作为急性排斥反应早期诊断的预见性和特异性指标.     

同系或同种胸腺与异种胸腺混合移植诱导T细胞免疫耐受的实验研究

目的 探讨同系或同种胸腺与异种胸腺混合移植诱导移植免疫耐受而不发生自身免疫性疾病的可行性。方法 BALB／c裸小鼠的肾包膜下植入胸腺建立异种、同系与异种混合、同种与异种混合胸腺移植模型。4个月后，用流式细胞仪技术检测受者裸小鼠外周血中CD3^＋CD4^＋T细胞，用单向混合淋巴细胞培养了解受者T细胞对ConA以及同种异种淋巴细胞的反应性，异基因皮肤移植观察皮肤移植物存活期。用ELISA法检测受体血清中抗DNA自身抗体。结果 胎胸腺移植4个月后，受者裸小鼠中CD3^＋CD4^＋T细胞重建良好；T细胞对ConA以及无关的SD鼠异种淋巴细胞的反应性强，而对移植胸腺供体来源的淋巴细胞不发生增殖反应；受者对移植胸腺供体来源鼠皮肤移植物不发生排斥，而对无关的SD鼠皮肤移植物发生排斥。异种胸腺移植组发生消耗性疾病死亡高，抗DNA自身抗体明显高于混合胸腺移植组。结论 同系或同种异种胎胸腺混合移植可诱导受者T细胞对供者移植物的免疫耐受。     

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.