雷帕霉素抑制小鼠骨髓移植后移植物抗宿主病作用的初步研究

目的：探讨新型免疫抑制药物雷帕霉素（RAPA）对小鼠异基因骨髓移植（allo-BMT）后急性移植物抗宿主病（aGVHD）的抑制作用，并与传统免疫抑制药物比较作用效果。方法：用主要组织相容性抗原（MHC）完全不合的纯种近交系小鼠[供鼠为雄性c57BL／6J（H-2“）鼠，受鼠为雌性BALB／c（H-2^d）鼠]建立allo-BMT／aGVHD动物模型，随机分5组，第1、2和3组分别给予RAPA、环孢菌素A（CsA）加甲氨蝶呤（MTX）、RAPA加CsA加MTX联合用药作为aGVHD抑制方案，第4组为空白对照，第5组为未移植组。观察各组小鼠骨髓移植后aGVHD的出现情况。结果：移植小鼠出现典型的aGVHD症状，未用aGVHD预防方案的移植小鼠（第4组）死亡高峰在移植后第5～7天，死亡率达100％。用不同aGVHD预防方案的第1～3组小鼠aGVHD症状明显减轻，平均生存时间（MST）较第4组显著延长（分别延长3．5、1．9和6．0d，P〈0．05）。不同aGVHD预防方案的移植小鼠之间，移植后外周血常规变化差异无显著性。经用各预防方案后，移植小鼠aGVHD病理表现减轻，且RAPA、CsA和MTX联合用药组的病理分级轻于RAPA单药组或CsA加MTX标准方案组。结论：RAPA可以显著抑制体外T淋巴细胞增殖效应，增强小鼠对骨髓移植的免疫耐受，对小鼠骨髓移植后移植物抗宿主病（GVHD）有明显抑制作用，可减轻GVHD症状和病理损害程度，延长平均生存时间。     

白介素-11对小鼠异基因骨髓移植后aGVHD的预防作用

目的 评估白介素-11(Interleukin-11,IL-11)对小鼠异基因骨髓移植(allo-BMT) 后急性移植物抗宿主病(aGVHD) 的预防作用. 方法 用主要组织相容性抗原(MHC) 完全不合的纯种近交系SPF级小鼠,雄C57BL/6(H-2b)为供鼠,雌性BALB/C(H-2d)为受鼠,建立allo-BMT/aGVHD 动物模型,随机分3组:模型对照组、IL-11组(剂量为500μg/(kg·d)、环孢菌素A(CsA)组(剂量为10mg/(kg·d),观察其对aGVHD 的预防作用. 结果 IL-11组、CsA组受鼠生存时间均长于模型对照组(P<0.01);IL-11组、CsA组受鼠aGVHD临床表现和病理改变均较模型对照组明显轻.IL-11组、CsA组之间平均生存时间比较无统计学差异(P=0.901). 结论 IL-11可有效预防小鼠allo-BMT后aGVHD,减轻其症状和病理损害程度,显著延长平均生存时间.     

树突状细胞亚群比例对小鼠异基因骨髓移植后急性移植物抗宿主病的影响

目的 探讨树突状细胞(Dendridc cell，DC)亚群比例对小鼠异基因骨髓移植(allo-BMT)后急性移植物抗宿主病(aGVHD)的影响。方法用主要组织相容性抗原(MHc)完全不合的近交系小鼠建立小鼠allo-BMT的模型，随机分为三组，分别在移植前、后和不对供鼠使用重组小鼠粒系集落刺激因子rmG-CSF，移植后均采用环孢素A(CsA) 甲氨蝶呤(MTX)方案预防aGVHD，观察三组小鼠移植前后外周血中DC亚群的比例及变化以及aGVHD的情况。结果 移植前后使用rhG-CSF均可以使DC亚群发生改变。使DC2的比例增加，移植前使用更明显；处理组移植小鼠aGVHD严重程度明显比不使用rmG-CSF组弱。移植前使用aGVHD表现最轻。结论 移植前后使用rmG-CSF可以使小鼠DC亚群比例发生改变，使DC2比例增高，并可使减轻移植小鼠aGVHD的症状。     

来氟米特预防小鼠异基因骨髓移植后aGVHD的实验研究

目的 评估来氟米特（LEF）对小鼠异基因骨髓移植（allo-BMT）术后急性移植物抗宿主病（aGVHD）的预防作用.方法 以SPF级雄性C57BL/6（H-2b）为供鼠,雌性BALB/C（H-2d）为受鼠,建立allo-BMT/aGVHD动物模型,并将大鼠随机分为：A组（模型对照组）、B组（环孢菌素A组,剂量为10 mg·kg^-1·d^-1）、C组（LEF组,剂量为10 mg·kg^-1·d^-1）,同时根据临床表现、病理组织学检查、平均生存时间评估各组aGVHD的严重程度.结果 B组和C组受鼠平均生存时间均较A组显著延长（均P〈0.01）,B组和C组间平均生存时间的差异均无统计学意义（均P〉0.05）;B组和C组受鼠的aGVHD临床表现和病理改变均较A组明显减轻.结论 LEF可有效预防小鼠allo-BMT术后出现的aGVHD,并可减轻其症状和病理损害程度,同时显著延长平均生存时间.     

霉酚酸脂联合环孢素和甲氨蝶呤预防外周血干细胞移植后急性移植物抗宿主病

目的　比较环孢素A(CsA)和短程甲氨蝶呤 (MTX)加或不加用短程霉酚酸脂 (MMF)预防急性移植物抗宿主病 (aGVHD)的效果。方法　人类白细胞抗原 (HLA)相合的异基因外周血造血干细胞移植 (allo PBSCT) 39例 ,常规环磷酰胺和全身照射为主进行预处理。GVHD预防分二组 :CsA +MTX组 2 6例 ,应用小剂量CsA(2mg·kg-1·d-1)和短程MTX(分别在移植后 1、3、6和 11d) ;MMF +CsA+MTX组 13例 ,在CsA和MTX基础上加用短程MMF 2g/d ,移植后口服 1～ 2 8d ,但MTX只在移植后1、3、6d应用。结果　两组患者移植后均顺利重建造血 ,中性粒细胞和血小板恢复时间的差异无显著意义 (P >0 0 5 ) ;MMF +CsA +MTX组aGVHD的发生率为 7 6 % ,未见II度以上aGVHD ,明显低于CsA+MTX组 (46 2 % ,P <0 0 5 ) ,CsA +MTX组II度以上aGVHD为 2 3 0 %。MMF +CsA +MTX组重度粘膜炎发生率 (15 4 % )亦明显低于CsA +MTX组 (30 8% )明显减少。结论　短程MMF联合CsA和短程MTX方案对allo PBSCT后的aGVHD的预防效果明显优于常规CsA和短程MTX ,其对慢性GVHD和移植后复发率的影响有待于进一步随访观察     

阿扎胞苷和CsA＋MTX对小鼠Allo—PBSCT后急性GVHD的预防效果比较

摘要：目的比较阿扎胞苷（AzaC）和环孢素A＋甲氨蝶呤（CsA＋MTx）对小鼠异基因外周血干细胞移植（A110～PBSCT）后发生急性移植物抗宿主病（aGVHD）的预防效果并初步分析其原因。方法雄性SPF级BAI。B／c（H-2d）小鼠为供鼠，雌性C57BI。／6（H-2b）小鼠为受鼠，随机选20只雌性C57BL／6（H-2b）受鼠为预处理组。另将供、受鼠随机分为PBS组、AzaC组和CsA＋MTX组各40只（每组含雄性供鼠、雌性受鼠各20只）。给PBS组、AzaC组和CsA＋MTX组的受鼠全身放疗（TBI）后输注供鼠T细胞去除（TCD）的外周血单个核细胞（PBMNC）和T细胞，再皮下注射PBS、AzaC和CsA＋MTX。检测受鼠的嵌合状态、生存天数、aGVHD观察及平均体重变化百分率和外周血CD4＋CD25＋FOXP3＋Tregs百分率。结果PBS组、AzaC组和csA＋MTx组受鼠外周血白细胞中均检测到雄性供鼠的sRY基因，未移植组＋20d内相继死亡，PBS组较早出现较严重的aGVHD，CsA＋MTX组随后也出现了aGVHD，而AzaC组未出现aGVHD。AzaC组的平均体重变化百分率、生存天数和外周血CD4＋CD25＋FOXP3＋Tregs百分率均与CsA＋MTX组和PBS组差异有统计学意义（P〈O．01）。结论AzaC预防小鼠A110—PBSCT后aGVHD的效果优于CsA＋MTX，可能与其增加了移植后外周血cD4＋CD25＋FOXP3＋Tregs数量有关。     

ATG用于异体造血干细胞移植前预防移植物抗宿主反应

目的 分析异体造血干细胞移植前加用抗T淋巴细胞球蛋白(ATG)预防移植物抗宿主反应(GVHD)的疗效.方法 27例白血病患者分为两组,每组移植供体来源及病种类似,分别有同胞兄妹、无关供体和人类白细胞抗原(HLA)半相合供体.A组(n=12):采用经典的CsA+MTX预防GVHD;B组(n=15):采用CsA + MTX+ATG预防GVHD.结果 B组15例患者全部存活,除4例出现Ⅱ°急性GVHD(aGVHD)外,其余11例均在移植后30 d左右仅出现Ⅰ° GVHD,并很快控制.A组12例患者中,3例HLA半相合移植患者分别在移植后第7、9和10天即出现Ⅳ°超急性GVHD,另有1例无关供体患者出现Ⅲ° aGVHD,该4例患者由于重度GVHD继发肺部感染而死亡;其余8例同胞之间的移植分别出现Ⅱ°～Ⅲ°aGVHD.结论 在异体造血干细胞移植前应用ATG可以有效预防GVHD的发生或减轻GVHD的严重度,明显减少移植相关死亡率.     

ATG用于异体造血干细胞移植前预防移植物抗宿主反应

目的分析异体造血干细胞移植前加用抗T淋巴细胞球蛋白(ATG)预防移植物抗宿主反应(GVHD)的疗效。方法27例白血病患者分为两组,每组移植供体来源及病种类似,分别有同胞兄妹、无关供体和人类白细胞抗原(HLA)半相合供体。A组(n=12):采用经典的CsA MTX预防GVHD;B组(n=15):采用CsA MTX ATG预防GVHD。结果B组15例患者全部存活,除4例出现Ⅱ°急性GVHD(aGVHD)外,其余11例均在移植后30 d左右仅出现Ⅰ°GVHD,并很快控制。A组12例患者中,3例HLA半相合移植患者分别在移植后第7、9和10天即出现Ⅳ°超急性GVHD,另有1例无关供体患者出现Ⅲ°aGVHD,该4例患者由于重度GVHD继发肺部感染而死亡;其余8例同胞之间的移植分别出现Ⅱ°~Ⅲ°aGVHD。结论在异体造血干细胞移植前应用ATG可以有效预防GVHD的发生或减轻GVHD的严重度,明显减少移植相关死亡率。     

高压氧对急性移植物抗宿主病小鼠细胞因子表达的影响

目的:观察高压氧(HBO)对骨髓移植时急性移植物抗宿主病(aGVHD)小鼠脾组织中细胞因子表达的影响,探讨HBO抗aGVHD的作用机制.方法:应用直线加速器排空C57BL/6小鼠骨髓后,注入BALB/C小鼠骨髓与脾淋巴细胞的混合液,复制aGVHD模型.应用RT-PCR技术检测经HBO、环孢素A(CsA)、氨甲喋呤(MTX)处理后,小鼠脾组织中白细胞介素2(IL-2)、肿瘤坏死因子α(TNF-α)、白细胞介素4(IL-4)及白细胞介素10(IL-10)mRNA水平的变化.结果:HBO组小鼠11d的生存率明显高于异基因移植组及CsA MTX组;HBO组小鼠脾组织中TNF-α和IL-2mRNA相对含量显著低于异基因移植组(P＜0.05)、显著高于CsA MTX组;HBO组小鼠脾组织中IL-4和IL-10 mRNA相对含量显著高于异基因移植组及CsA MTX组(P＜0.05).结论:HBO可能通过调节促炎/抑炎细胞因子的分泌而发挥其明显的抗aGVHD作用.     

自然杀伤细胞对小鼠异基因骨髓移植中移植物抗宿主病效应的影响

目的:研究自然杀伤细胞(NK)在异基因骨髓移植中对移植物抗宿主病(GVHD)的影响.方法:以近交系小鼠C57BL/6(H-2b)为供鼠、BALB/c(H-2d)为受鼠,在移植物中增加供者的外周T细胞和(或)NK细胞进行异基因骨髓移植,根据临床表现和病理检查,比较不同移植组的存活率和GVHD发生情况.结果:增加1×106供鼠NK细胞的移植组中40%小鼠出现GVHD,增加2×106供鼠NK细胞的移植组中仅30%出现GVHD;与单纯异基因骨髓移植细(80%急性GVHD、20%慢性GVHD)和骨髓 T细胞组(100%急性GVHD)比较,GVHD发生率显著下降(P＜0.01),30、60和120 d存活率显著增高(P＜0.01).结论:在小鼠异基因骨髓移植中,同种异基因反应性NK细胞可抑制GVHD效应,延长存活期.     

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.