人脐血造血祖细胞体外培养的研究

应用体外培养方法对人脐血造血祖细胞（ＣＦＵ－ＧＭ、ＣＦＵ－Ｅ、ＢＦＵ－Ｅ、ＣＦＵ－Ｆ）数量及其特征进行观察，并与成人外周血及骨髓相比较。结果显示：①脐血中ＣＦＵ－ＧＭ、ＣＦＵ－Ｅ、ＢＦＵ－Ｅ数量约相当于成人外周血的７～１８倍；ＣＦＵ－ＧＭ及ＣＦＵ－Ｅ数量与骨髓相比无统计学差异，但ＢＦＵ－Ｅ低于骨髓；②脐血中ＣＦＵ－ＧＭ的集落大小、细胞构成、集落生长曲线明显不同于成人外周血及骨髓；③脐血红系祖细胞生长模式近似于骨髓；④脐血中可培养出ＣＦＵ－Ｆ，其数量约为骨髓的２／５，而外周血中则无ＣＦＵ－Ｆ形成。提示人脐血中富含造血干／祖细胞，而且在数量和特征上明显不同于外周血和骨髓，在移植中可能更具有重建骨髓造血的能力。     

脐血造血干细胞特性研究

应用造血祖细胞体外培养和流式细胞术（ＦＣＭ），观察了脐血中粒一单系祖细胞（ＣＰＵ－ＧＭ）、红系祖细胞（ＢＦＵ－Ｅ）的数量和特性，并对其细胞增殖周期和骨贿及成人外周血做了比较，实验提示脐血中含丰富的ＣＦＵ－ＧＭ和ＢＦＵ－Ｅ，其数量低于成人骨髓而大大超过外周血水平，脐血红系祖细胞对ＥＰＯ、ＢＰＡ表现出更高的敏感性，而粒系祖细胞对ＣＳＦ的敏感性和骨髓比较似无明显差别．脐血Ｓ期、Ｇ２＋Ｍ期，及Ｓ＋Ｇ２Ｍ期细胞数均明显低于骨髓，虽略高于外周血各期细胞数，但差别无显著性意义．     

人脐血粒单系及在纤维系祖细胞的体外培养

采用体外单层半固体琼脂培养法及液体培养法检测了脐知中ＣＦＵ－ＧＭ和ＣＦＵ－Ｆ并与成人骨髓和外周血对照，结果显示，脐血培养７ｄＣＦＵ－ＧＭ产率约相当于成人外周血的５～６倍，但低于骨髓；培养１４ｄ的产率略高于骨髓，与骨髓培养７ｄ结果相比无血中可培养出ＣＦＵ－Ｆ，但产率仅为骨髓的１／５，而外周血中则无ＣＦＵ－Ｆ形成，提示人脐血富含造血祖细胞，且有不同于骨髓及外周血的性质。     

细胞因子及4℃保存对脐血造血干／祖细胞的影响

应用体外半固体培养方法,研究了粒巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）、白细胞介素３（ＩＬ－３）、白细胞介素６（ＩＬ－６）和促红细胞生成素（Ｅｐｏ）的不同组合及４℃保存对脐血造血干／祖细胞ＣＦＵ－ＧＭ和ＨＰＰ－ＣＦＣ产率的影响。结果表明：在琼脂半固体培养体系中,在不同组合的细胞因子作用下,脐血ＣＦＵ－ＧＭ产率有是明显差异;以ＧＭ－ＣＳＦ组最低、ＧＭ－ＣＳＦ＋ＩＬ－３＋ＩＬ－６＋Ｅｐｏ组最高。脐血置４     

重组白细胞介素2对骨髓造血祖细胞体外增殖的影响

为了了解重组白细胞介素２对骨髓造血祖细胞体外增殖的影响，应用体外半固体培养法研究了ｒｈＩＬ－２及其与ＧＭ－ＣＳＦ和Ｅｐｏ组合对骨髓祖细胞集落形成的调节作用。结果：ｒｈＩＬ－２单独对骨髓造血祖细胞的体外增殖无刺激作用；当其与ＧＭ－ＣＳＦ联合应用时，在浓度为１０．０×１０４～４０．０×１０４Ｕ·Ｌ－１时，ＣＦＵ－ＧＭ集落数明显高于ＧＭ－ＣＳＦ对照组；当其与Ｅｐｏ联用时，在浓度为１０．０×１０４～４０．０×１０４Ｕ·Ｌ－１时ＣＦＵ－Ｅ和ＢＦＵ－Ｅ集落数明显高于Ｅｐｏ单用组；但当其浓度升至１０．０×１０５Ｕ·Ｌ－１时，各种集落数均明显减少。重组人白介素２（ｒｈＩＬ－２）对骨髓造血祖细胞体外增殖的研究结果显示，在一定的浓度范围内，ｒｈＩＬ－２与ＧＭ－ＣＳＦ和Ｅｐｏ联合对骨髓造血祖细胞的体外增殖具有协同的刺激作用，单独应用无集落形成能力，大剂量应用可能会抑制骨髓造血。     

脐带血,成人外周血的干祖细胞检测及CFU—GM与CD34^＋细胞百分率？…

本文利用桥联酶标ＡＰＡＡＰ技术，用ＣＤ３４＾＋单抗ＱＢＥＮＤ１０标记脐血和外周血造血干细胞，同时采用体外半固体培养法检测脐血和外周血ＣＦＵ－ＧＭ产率血，并对两者的相关性进行了分析。结果表明：（１）体外培养ＣＦＵ－ＧＭ产率，脐血为９６．０６±６３．３７，外周血为７．２４±６．２０，两者差异显著，Ｐ〈０．０１。（２）ＣＤ３４＾＋细胞百分率，脐血为１．６９±１．０６％，外周血为０．００６５＋０．００４７     

脐带血、成人外周血的干祖细胞检测及CFU-GM与CD34~+细胞百分率相关性研究

本文利用桥联酶标ＡＰＡＡＰ技术，用ＣＤ３４＋单抗ＱＢＥＮＤ１０标记脐血和外周血造血干细胞，同时采用体外半固体培养法检测脐血和外周血ＣＦＵ－ＧＭ产率血，并对两者的相关性进行了分析。结果表明：（１）体外培养ＣＦＵ－ＧＭ产率，脐血为９６．０６±６３．３７，外周血为７．２４±６．２０，两者差异显著，Ｐ＜０．０１。（２）ＣＤ３４＋细胞百分率，脐血为１．６９±１．０６％，外周血为０．００６５＋０．００４７％，两者差异显著，Ｐ＜０．０１。（３）脐血ＣＦＵ－ＧＭ产率与ＣＤ３４＋细胞百分率呈正相关，Ｒ＝０．７５９，Ｐ＜０．０１，回归方程Ｙ＝０．０１２７Ｘ＋０．４６４９。外周血不相关Ｒ＝０．０４４７，Ｐ＜０．０５。提示采用桥联酶标法检测ＣＤ３４＋细胞百分率可以代替ＣＦＵ－ＧＭ用于预测脐血造血重建能力，由于该法简便、快速、敏捷，不需昂贵设备，值得推广和应用     

细胞因子及4℃保存对脐血造血干/祖细胞的影响

应用体外半固体培养方法,研究了粒巨噬细胞集落刺激因子（ＧＭＣＳＦ）、白细胞介素３（ＩＬ３）、白细胞介素６（ＩＬ６）和促红细胞生成素（Ｅｐｏ）的不同组合及４℃保存对脐血造血干／祖细胞ＣＦＵＧＭ和ＨＰＰＣＦＣ产率的影响。结果表明：在琼脂半固体培养体系中,在不同组合的细胞因子作用下,脐血ＣＦＵＧＭ产率有明显差异;以ＧＭＣＳＦ组最低,ＧＭＣＳＦ＋ＩＬ３＋ＩＬ６＋Ｅｐｏ组最高。脐血置４℃保存４ｄ后,ＣＦＵＧＭ和ＨＰＰＣＦＣ产率明显降低。     

低温冷冻对脐血CD＋34细胞和造血祖细胞增殖的影响

采用－８０℃低温冷冻法,观察冷冻前及冷冻后１、２、３周时单个核细胞（ＭＮＣ）的回收率,脐血造血前体细胞在体外的增殖力,并用流式细胞仪检测ＣＤ３４＋细胞的百分率。旨在探讨低温冷冻对脐血造血前体细胞的影响。结果表明：脐血冷冻后１、２、３周时ＭＮＣ的回收率分别为９３．６％,９１．５％和９０．７％（Ｐ＞０．０５）;细胞活力分别为９８％,９７％,９５％和９１％（Ｐ＞０．０５）;ＣＤ３４＋细胞依次为１．２０％、１．１８％、１．１６％和１．１７％;ＣＦＵ－ＧＭ,ＣＦＵ－Ｅ冷冻后１、２周时回收率达９７％和９５％（Ｐ＞０．０５）,第３周时达８９％（Ｐ＞０．０５）。结果提示：长期冷冻可能导致冷冻之脐血祖细胞产率下降,但对代表造血干细胞的ＣＤ３４＋细胞影响则不大。     

脐血血清对正常及白血病骨髓粒－单祖细胞生长的影响

利用粒－单祖细胞集落（ＣＦＵ－ＧＭ）的体外培养，观察脐血血清（ＵＣＳ）对正常及白血病骨髓造血祖细胞生长的影响，结果表明：ＵＣＳ对正常及急性白血病（ＡＬ）患者完全缓解的骨髓ＣＦＵ－ＧＭ形成有一定增殖作用，而对初发／复发未治ＡＬ患者的骨髓ＣＦＵ－ＧＭ形成无类似作用。     

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.