昆布多糖对大鼠骨髓巨噬细胞生物活性的影响

目的 研究昆布多糖对大鼠骨髓巨噬细胞生物活性的影响.方法 制备并体外培养大鼠骨髓造血干/祖细胞、巨噬细胞;制备昆布多糖作用后的骨髓巨噬细胞条件培养液,测定培养液中造血生长因子促红细胞生成素(EPO)、粒-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-3(IL-3)表达及活性;测定昆布多糖条件培养液作用后造血祖细胞的集落产率.结果 经昆布多糖诱导的骨髓巨噬细胞培养上清液可显著提高骨髓造血祖细胞的集落产率;经昆布多糖诱导后,骨髓巨噬细胞的EPO、GM-CSF和IL-3的表达增高.结论 昆布多糖可刺激骨髓巨噬细胞造血因子表达增高,从而促进髓系多向性造血祖细胞(CFU-Mix)、红系祖细胞(CFU-E)、粒一单系造血祖细胞(CFU-GM)的增殖分化.     

造血生长因子对脐血单个核细胞的体外增殖效应

目的 :探讨不同造血生长因子组合对人脐血造血细胞的体外增殖效应 .方法 :采用免疫荧光染色和祖细胞集落测定 ,观察人脐血单个核细胞 (MNC)经多种造血生长因子的不同组合作用 2wk后其有核细胞、CD34+ 细胞和祖细胞数量的变化 ,并用脾结节形成实验和骨髓祖细胞培养等观察其在小鼠体内的短期造血能力 .结果 :脐血MNC分别与A(SCF /IL 3/IL 6 /G CSF/GM CSF) ,B(SCF/IL 3/IL 6 /FL/EPO) ,C(SCF/IL 3/IL 6 /EPO ) ,3种造血生长因子组合悬浮培养 2wk ,有核细胞、CD34+ 细胞和祖细胞数量增加 .有核细胞总数在B组因子作用下增加最显著 [(2 6± 5 )倍 ],而CD34+ 细胞数量在A组因子作用下增加最明显 [(8± 2 )倍 ].3种因子组合对不同种类祖细胞的增殖效应不同 ,其中A组因子主要刺激早期混合系祖细胞CFU Mix扩增 [(11± 2 )倍 ],B组因子主要刺激粒单系祖细胞CFU GM扩增 [(2 2± 3)倍 ],C组因子主要刺激早期红系祖细胞BFU E扩增 [(13± 4 )倍 ].此外 ,给受照小鼠输注A组因子作用后的人脐血细胞 ,13d在小鼠脾脏形成脾结节 (CFU S) ,30d从小鼠骨髓检出造血祖细胞(CFU GM和CFU E) ,造血细胞数量与从输注新鲜脐血细胞的小鼠体内检出的结果无显著性差异 (P >0 .0 5 ) .结论 :造血细胞生长因子对人脐血单个核细胞具有体外     

脐血造血干细胞体外培养及其特性研究

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-单系祖细胞(CFU-GM)和红系祖细胞(BFU-E)的数量和特性,并对其细胞增殖周期和骨髓及成人外周血做了比较.结果表明,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;脐血红系祖细胞对促红细胞生成素(EPO)和爆式促进活性(BPA)表现出更高的敏感性,而粒系祖细胞对集落刺激因子(CSF)的敏感性和骨髓比较似无明显差别;脐血S期、G_2+M期及S+G_2M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.提示脐血造血干细胞体外培养可以得活性较高的祖细胞.     

小鼠腹腔巨噬细胞对红系祖细胞增殖的影响

应用造血祖细胞体外培养技术,观察了小鼠腹腔巨噬细胞对早期红系祖细胞(BFU-E)及晚期红系祖细胞(CFU-E)增殖的影响。结果表明,小鼠腹腔贴壁巨噬细胞对CFU-E表现为刺激作用;对BFU-E增殖依培养体系条件不同而表现为刺激和抑制双重作用。巨噬细胞培养上清液也表现相似的作用。提示巨噬细胞具有促红细胞生成素(EPO)和爆式集落促进活性(BPA)样活性。     

烧伤后血清中造血刺激活性组分分析

目的　检测烧伤血清中是否有异常的造血刺激活性组分出现,并对其性质作初步鉴定。方法　采用离子交 换层析分离正常及烧伤血清,用CFU -E、CFU- GM集落培养法检测各分离组分在烧伤前后造血刺激活性的变化,并对其进 行热、酸、碱和酶处理后进一步检测造血活性。结果　小鼠烧伤血清经离子交换层折后得到A、B、C、D4个组分,其中A、B 组分红系造血刺激活性增高;C组分对红系和粒系造血刺激活性增高;且活性最强。D组分不具有造血刺激活性。A、B、C 3组分经酸处理和胰酶消化后活性消失,A、B组分经碱处理和热处理后活性消失,而C组分加热70℃以及经碱处理后仍 保持部分活性。结论　①烧伤后血清经离子交换色谱分离得到的A、B、C、D4组分中,A、B组分具有红系刺激活性,C组 分活性最强,对红系和粒系均有刺激。②A、B、C3个活性组分均为蛋白质,C组分对酸不稳定,对热和碱具有一定的稳定 性,而A、B两组分既不耐热,对酸碱均不稳定。     

当归多糖对骨髓巨噬细胞的影响及其与造血调控的关系

目的研究当归多糖(APS)对骨髓巨噬细胞(BMMФ)生物活性的影响及其与造血调控的关系，从而阐明当归的活血机制。方法采用骨髓造血祖细胞和巨噬细胞体外培养，造血生长因子生物活性检测，免疫细胞化学，核酸探针原位杂交等技术。结果经APS诱导的BMMФ的培养上清液可提高髓系造血祖细胞的集落产率；经APS诱导后，BMMФ表达促红细胞生成素(EPO)、粒一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)的蛋白水平有不同程度的提高；EPO mRNA和GM-CSF mRNA的表达水平和强度明显提高。结论APS可以直接和(或)间接地刺激造血诱导微环境中的BMM中，从基因水平和蛋白水平上促进造血调控因子的合成和分泌，进而促进髓系多向性造血祖细胞(CFU-Mix)、晚期红系祖细胞(CFU-E)、粒单系造血祖细胞(CFU-GM)的增殖分化。     

促红细胞生成素激活新的细胞信号途径发挥心脏保护作用

促红细胞生成素（erythropoietin，EPO）主要是由肾脏间质细胞（少量来自肝脏）合成和分泌的含唾液酸的糖蛋白生长因子。传统观点认为．EPO只作用于骨髓中造血干／祖细胞和幼稚红细胞表面特异性促红细胞生成素受体（EPOreceptor．EPOR），刺激造血前体细胞增殖和成熟．抑制造血前体细胞的凋亡．促使红细胞自骨髓向血液释放…。重组人促红细胞生成素（recombinant human EPO，rhEPO）也仅用于慢性贫血如肾性贫血、HIV相关贫血、恶性肿瘤性贫血和自体输血的血液储存。尽管有研究观察到EPO对贫血伴心力衰竭患者的心功能有所改善．但因其可能增加血栓形成和高血压的发病率．从而限制了EPO在心血管疾病中的应用。近来发现．EPOR在多种非红系细胞包括神经细胞、心肌细胞、血管内皮细胞以及成肌细胞表面表达，EPO及其氨甲酰化衍生物（carbamylated EPO，CEP0）对神经系统和心血管系统也具有强大的保护作用。     

贫血患者瘦素水平的测定

目的　通过对贫血患者体内瘦素水平的测定 ,了解在贫血状态下是否存在瘦素分泌的异常。方法　分别抽取正常人、再生障碍性贫血 (再障 )、营养不良性贫血、溶血性贫血患者骨髓液及外周血 ,用放射免疫分析法分别测定其瘦素含量。结果　营养不良性贫血、溶血性贫血患者骨髓瘦素含量和外周血瘦素含量比 (BM/PB)与正常人接近 (P >0 0 5 ) ,而再障患者两者的比值比正常人明显升高 (P <0 0 1 ) ,而贫血程度与瘦素含量无明显相关性 (P >0 0 5 ) ,但骨髓与外周血瘦素含量则高度相关 (P <0 0 1 )。结论　再障患者瘦素水平明显高于正常人及其他类型贫血患者 ,可能与再障骨髓脂肪细胞明显增多、脂肪细胞分泌瘦素明显增多有关。瘦素受体的不完整及造血干 /祖细胞对干细胞因子 (SCF)的反应性降低、破坏了瘦素与SCF协同刺激原始造血祖细胞增殖的作用 ,可能是造成再障瘦素水平高 ,但造血干 /祖细胞数量减少的一个原因     

黄芪注射液对贫血小鼠巨核系造血的作用及其机理的研究

目的　研究黄芪注射液 (AMI)对贫血小鼠巨核系造血的影响及其作用机制。方法　制备贫血小鼠模型 ,随机分为给药组和对照组 ,分别腹腔注射 AMI〔2 0 m g/ (ml· 2 0 g)〕或等量生理盐水 q.d× 6 ,于给药后第8、11、14天常规方法检测外周血血小板数、骨髓有核细胞数 ,同时用体外巨核系祖细胞 (CFU- Meg)培养方法检测CFU- Meg,采集血清作巨核细胞系集落刺激活性 (Meg- CSA)测定。结果　贫血小鼠给药组和对照组比较 ,血清Meg- CSA明显升高 ,给药后第 11天所测各项指标均恢复正常 ,恢复时间明显缩短。结论　 AMI能够升高血清Meg- CSA,加速骨髓抑制后巨核系造血功能的恢复     

用胚胎干细胞诱导的造血干/祖细胞重建小鼠造血功能研究

目的 胚胎干细胞体外诱导分化出现CD34^ 造血干/祖细胞，然后将其注射入致死量照射小鼠体内，以研究其重建造血功能。方法 胚胎干细胞自由分化形成胚胎体后，加入造血刺激因子混合液以诱导产生CD34^ 造血干／祖细胞，将这些造血干／祖细胞注射入经致死剂量照射的小鼠，观察小鼠存活率的改变，并取存活2个月的小鼠骨髓和脾脏，PCR检测嵌合体形成。结果 体外分化第13日CD34^ 造血干／祖细胞比例可高达17．36％，将这些细胞注射入致死量照射小鼠后，可提高存活率达86．67％，存活小鼠的骨髓和脾脏均检测到供体来源的细胞。结论 胚胎干细胞体外分化获得CD34^ 造血干/祖细胞，且后者具有其相应的正常生物学功能。     

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.