化疗+G-CSF方法动员外周血干细胞时骨髓及外周血相关细胞亚群的动态变化

目的探讨应用化疗 G-CSF方法动员外周血干细胞时,恶性血液病患者的骨髓及外周血CD34 细胞和T细胞亚群的变化特点.方法 16例拟行自体外周血干细胞移植患者,在应用化疗 G-CSF方法动员外周血干细胞期间,应用流式细胞仪测定骨髓及外周血CD34 和T细胞亚群动态变化情况.结果 1.恶性血液病患者在应用G-CSF前,骨髓中CD34 细胞的初始值为(0.52±0.31)%.应用G-CSF 48 h后,CD34 细胞测定值为(1.22±0.42)%,较前有明显增加(P＜0.001).CD34 细胞的高峰值出现在应用G-CSF 96 h后,为(2.23±0.34)%,较未应用G-CSF时差异显著(P＜0.001).2.恶性血液病患者在应用G-CSF前,外周血CD34 细胞测定值为(0.39±0.27)%.在应用G-CSF 72 h后外周血中CD34 细胞的测定值为(1.29±0.64)%,较前有明显增加(P＜0.001).应用G-CSF96 h后,CD34 细胞的测定值达高峰,为(1.41±0.73)%,较未应用G-CSF前差异显著(P＜0.001).3.无论是否应用G-CSF,恶性血液病患者的T淋巴细胞亚群比例均处于倒置状态,应用G-CSF后随CD34 细胞的逐渐增加,T淋巴细胞亚群变化不明显(P＞0.05).结论恶性血液病患者,在应用化疗 G-CSF动员外周血干细胞时,其骨髓及外周血CD34 细胞和T细胞亚群的动态变化各有其特点.     

G-CSF动员的外周血与骨髓CD34+细胞黏附分子的表达

目的：研究黏附分子在外周血干细胞动员机制中的作用。 方法：应用流式细胞仪检测粒细胞集落刺激因子G-CSF动员的外周血细胞与骨髓CD34⁺细胞表达的黏附分子非常延迟抗原4(VLA-4)、淋巴细胞功能相关抗原1(LFA-1)和L-选择素(CD62L)。 结果：G-CSF动员后的外周血CD34⁺细胞表达的VLA-4、LFA-1与骨髓CD34⁺细胞相比明显减少,CD62L无明显改变。 结论：黏附分子VLA-4及LFA-1降低可能是G-CSF动员造血干细胞进入外周血过程中的一个重要环节。     

rhG-CSF动员对外周血淋巴细胞亚群和CD34+细胞的影响

目的:动态观察自体外周血造血干细胞移植患者经rhG-CSF动员过程中外周血及外周血造血干细胞(PBPC)中淋巴细胞亚群和CD34 细胞的变化,指导临床选择最佳采集时机。方法:对大剂量化疗(HDC)联合重组人粒细胞集落刺激因子(rhG-CSF)动员的38例白血病和淋巴瘤患者,采用流式细胞术(FACS)测定其动员前后外周血及PBPC中的淋巴细胞亚群CD3、CD4、CD8、NK、CD19和造血干细胞CD34细胞含量的变化,同时用甲基纤维素半固体培养基进行CFU-GM培养来评价干细胞克隆生成能力。结果:动员后外周血淋巴细胞亚群CD3、CD4细胞含量均低于动员前(P<0.01),而动员后外周血中的CD34细胞含量明显高于动员前(P(0.05)。动员后第5天CD34含量达到最高峰。PBPC中CD4、CD4/CD8明显低于外周血(P<0.005),其他淋巴细胞亚群含量与外周血比较无明显变化。动员后外周血中CD34 细胞明显高于动员后,动员后PBPC中CFU-GM生成明显高于外周血(P<0.05),CD4/CD8比值严重倒置,B细胞恢复较快。结论:大剂量化疗联合rhG-CSF动员会使外周血淋巴细胞亚群发生不同程度的变化并明显增加外周血造血干细胞含量。     

自体与异基因外周血干细胞移植早期造血重建的初步研究

目的探讨自体外周血干细胞移植(auto-PBSCT)患者与异基因外周血干细胞移植(allo-PBSCT)供者的动员后外周血(MPB)及产品中CD34 细胞及其亚群所占比例、数量及其在移植后造血重建中的意义。方法应用流式细胞仪分别检测16名auto-PBSCT患者和24名allo-PBSCT供者MPB及分离产品的115份标本,比较CD34 、CD34 CD38-细胞所占比例及两组患者早期造血恢复情况。结果allo-PBSCT供者MPB及分离产品中CD34 细胞比例均明显低于auto-PBSCT患者(P<0.05);MPB及分离产品中CD34 CD38-细胞亚群含量在两组人群中无显著差异(P>0.05);allo-PBSCT患者所接受的单核细胞数明显多于auto-PBSCT患者(P<0.01);两组患者移植后中性粒细胞恢复到0.5×109/L、血小板恢复到2×1010/L的时间无显著差异(P>0.05),但均与单位体质量输入的单个核细胞、CD34 、CD34 /CD38-细胞数量相关。结论无论是auto-PBSCT 还是allo-PBSCT,回输的CD34 、CD34 CD38-细胞数量是决定造血重建速度的关键因素。     

自体骨髓干细胞动员治疗右心衰竭的实验研究

目的:探讨粒细胞集落刺激因子(G-CSF)对野百合碱(MCT)诱导的大鼠右心衰竭(RHF)的治疗作用. 方法:动物随机分为3组:正常对照组(CON组)、MCT组、MCT/G-CSF组,腹腔注射野百合碱诱导大鼠右心衰竭模型,采用G-CSF动员自体骨髓干细胞(BMSC),动员结束后采集外周血标本,流式细胞仪检测外周血单个核细胞中CD34 细胞含量.第4周分别对3组大鼠进行血流动力学检测,处死大鼠,取肝、心脏、肺组织行苏木精-伊红(H-E)染色. 结果:①单个核细胞中CD34 细胞含量在MCT/G-CSF组明显高于MCT组、对照组,差异均具有统计学意义(P<0.01).②实验第35d,MCT组血流动力学指标明显高于对照组 (P<0.05),RHF造模成功;MCT/G-CSF组则低于MCT组(P<0.05),说明G-CSF干预治疗取得疗效.③MCT组大鼠右心室、肝脏、肺组织出现RHF的病理学改变,MCT/G-CSF组的病理学改变部分逆转. 结论:G-CSF能动员骨髓干细胞,部分逆转RHF的血流动力学和病理学改变、减缓RHF的发展,可以用于RHF的研究和治疗.     

粒细胞集落刺激因子动员正常供者外周血造血干细胞研究

目的:探讨粒细胞集落刺激因子(G-CSF)动员正常供者外周血造血干细胞的效果、影响因素及不良反应。方法:对20例造血干细胞正常供者采用中位剂量G-CSF10.9μg/(kg·d)皮下注射3～5天后,使用COBESpectra血细胞分离机采集外周血干细胞。采用流式细胞仪检测采集物中CD34 细胞数。结果:第一次采集的单个核细胞(MNC,个)及CD34 细胞量(个)分别为(1.12～13.06,中位数3.10)×108/kg供者体重及(1.14～10.42,中位数3.8)×106/kg供者体重。MNC及CD34 细胞采集总量分别为(2.21～13.60,中位数5.80)×108/kg受者体重及(2.30～14.00,中位数6.40)×106/kg受者体重,均达到移植要求。男性CD34 细胞总量显著高于女性。不良反应较为轻微。供者年龄、体重、G-CSF剂量、采集前白细胞数等与采集所得MNC及CD34 细胞量无明显相关性。结论:G-CSF作为正常供者动员剂安全有效。     

人脐带间充质干细胞对脐血CD34~+细胞在NOD/SCID小鼠体内造血重建的影响

目的 探讨人脐带间充质干细胞对脐血CD34~+细胞在NOD/SCID小鼠体内造血重建的影响.方法 将3.5×10~5个脐血CD34~+细胞单独(单移植组)或与5.0×10~6个人脐带间充质干细胞共同(共移植组)输入经~(137)Cs 3.0Gy照射后的NOD/SCID小鼠体内,观察移植后6周内小鼠外周血象的变化情况.于移植后第6周处死小鼠,采用流式细胞术检测小鼠骨髓、脾脏及外周血人源细胞(hCD45~+)含量,并分别检测小鼠骨髓中人源淋巴系(CD3/CD19)、粒系(CD33)、单核系(CD14)、血小板(CD61)、红系(CD235a)等各系血细胞比例,比较间充质干细胞共移植对CD34~+细胞植入率的影响.结果 移植后3周,两组小鼠外周血象开始有不同程度恢复;移植后6周,共移植组外周血白细胞和血小板计数均已达高峰,明显高于单移植组(P<0.05),两组小鼠的红细胞计数差异无统计学意义(P>0.05).移植后6周,共移植组骨髓及外周血中人源细胞hCD45~+CD34~+比例分别为(42.66±2.57)%和(4.74±1.02)%,明显高于单移植组的(25.27±1.67)%和(1.19±0.54)%(P=0.006).移植后6周,共移植组小鼠骨髓内的CD19~+、CD33~+、CD14~+、CD61~+和CD235a~+细胞比例均明显高于单移植组(P<0.05),CD3~+T淋巴细胞比例明显低于单移植组(P=0.003);CD19~+B淋巴细胞得到优势扩增,明显高于其他各系血细胞比例(P<0.05).结论 脐带间充质干细胞与脐血CD34~+细胞共移植可促进造血干细胞的植入,缩短CD34~+细胞移植后造血恢复时间.     

造血干细胞动员及采集的影响因素

目的 研究影响外周血干细胞的动员及采集因素.方法 35例血液病患者予以化疗+粒细胞集落刺激因子(G-CSF)动员,15例正常供者予以G-CSF动员.用Cobe Spectra血细胞分离机采集外周血干细胞.结果 外周血造血干细胞的有效动员与被动员者的年龄、性别、体表面积、采集的循环血量及骨髓是否曾经累及无关,而与动员所应用的化疗的种类相关;以环磷酰胺(CTX)为主的化疗+G-CSF动员优于柔红霉素+阿糖胞苷(DA)+G-CSF(P＜0.01);CTX为主的化疗+G-CSF动员产品单个核细胞与中剂量阿糖胞苷+G-CSF动员无统计学差异,CD34+含量则CTX组显著优于DA组(P＜0.05);中剂量阿糖胞苷+G-CSF动员与DA+G-CSF动员方案相比较,两组无统计学差异(P＞0.05).结论 外周血造血干细胞的有效动员与动员所应用的化疗种类相关,CTX为主的化疗+G-CSF动员效果最佳,蒽环类药物动员效果相对较差并动员前尽量减少使用.     

重组人G-CSF治疗不同白细胞减少症的疗效与机制

目的 探讨重组人粒细胞集落刺激因子(rhG-CSF)治疗不同白细胞减少症的疗效与机制.方法 临床选取四组患者: 再生障碍性贫血组(AA组,n=10)、骨髓增生异常综合征组(MDS组,n=10)、药物性白细胞减少组(药物组,n=20)(根据骨髓粒系增生程度再分为增生活跃组和增生减低组)和缺铁性贫血组(对照组,n=10).ELISA法测定外周血G-CSF浓度;流式细胞仪检测骨髓单个核细胞(BMMNC)G-CSF受体(G-CSFR)的表达率及CD34+细胞比例;培养BMMNC,比较粒细胞集落形成单位(CFU-G)集落数.比较AA组、MDS组和药物组进行rhG-CSF治疗的效果.结果 AA组和药物组的血清G-CSF浓度均高于对照组和MDS组(P<0.05).AA组的G-CSFR表达率低于其余各组(P<0.05),但MDS组与药物组和对照组之间差异无统计学意义(P>0.05);增生减低组G-CSFR表达率低于增生活跃组和对照组(P<0.05),但增生活跃组与对照组差异无统计学意义(P>0.05).AA组的CD34+细胞比例低于其余各组(P<0.05),但其余各组间差异无统计学意义(P>0.05).AA组和MDS组的CFU-G集落数均低于对照组和药物组(P<0.05),但AA组与MDS组、药物组与对照组之间差异无统计学意义(P>0.05).经rhG-CSF治疗后,药物组疗效优于MDS组,MDS组疗效优于AA组(P<0.05).结论 rhG-CSF疗效与骨髓G-CSFR表达率、CD34+细胞比例、干祖细胞对G-CSF的反应性及内源性G-CSF浓度有关.     

急性心肌梗死患者血浆VEGF、SDF-1和外周血CD34+细胞的动态改变及其意义

目的研究急性心肌梗死时血管内皮细胞生长因子(VEGF)、基质细胞衍生因子(SDF-1)以及外周血CD34 细胞的动态改变,探讨其在心肌梗死(心梗)中的作用。方法采集急性心梗患者发病第1、3、7、10、14天外周静脉血,应用酶联免疫方法检测心梗患者以及对照组患者血VEGF和SDF-1的水平。应用流式细胞仪检测心梗患者第1、7、14天外周血CD34 细胞的水平。同时对心梗患者进行心肌酶、肌钙蛋白、心电图及心脏超声等常规检查。结果(1)心梗后第7天外周血CD34 细胞(个/μl)明显增高(2.35±0.72vs1.48±0.49,P<0.05);(2)心梗后血VEGF(pg/ml)明显升高,于第14天达到高峰(197.56±39.87vs53.79±18.12,P<0.01);(3)心梗第1天血SDF-1(pg/ml)明显降低(1683.12±224.79vs2178.67±265.34,P<0.01),以后渐恢复至与对照组相同水平;(4)心梗第7天VEGF水平与外周血CD34 细胞高峰值之间有明显相关性;(5)VEGF高峰值与CK-MB高峰值和肌钙蛋白I之间均有明显相关性。结论心肌梗死本身即可动员干细胞至外周血;心梗后VEGF明显升高至少可持续2周以上,SDF-1短暂降低;两者的动态变化可能与干细胞动员及促使更多干细胞归巢至损伤心肌有关。     

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.