新改良外周血造血干细胞冷冻技术的临床应用观察

目的为临床自体造血干细胞移植提供一种简单易行、安全有效、成本低廉的干细胞冻存方法。方法 10份外周造血干细胞以二甲基亚砜、自体血浆作保护剂在-80℃条件下直接冻存,冻存前后检测外周血干细胞的活性、单个核细胞(MNC)数和CD34+细胞数及回收率。结果冻存前后外周血干细胞活性单个核细胞(MNC)数和CD34+细胞数及回收率满足移植要求,输注后造血均重建。结论二甲基亚砜、自体血浆作保护剂在-80℃条件下直接冻存干细胞是一种简便、可靠、安全、成本低的外周造血干细胞冻存方法。     

改良DICE方案联合自体造血干细胞移植治疗侵袭性非霍奇金淋巴瘤的效果观察

目的:探讨自体外周血造血干细胞移植中改良的DICE+G-CSF±R方案动员外周血干细胞以及用自体血浆代替人血白蛋白作为冻存保护液在非霍奇金淋巴瘤干细胞移植中的应用价值。方法:2例患者分别为间变大细胞淋巴瘤和伯基特淋巴瘤,在化疗达到完全缓解或部分缓解后,采用改良的DICE+G-CSF±R动员自体外周血干细胞,分别采集到干细胞122 mL和95 mL,单个核细胞(mononuclear cells,MNC)分别为3.28×109/kg和2.05×109/kg,CD34+细胞分别为3.5%、1.5%,采用羟乙基淀粉(hydroxyethyl starch,HES)、二甲基亚砜(dimethysulfoxide,DMSO)和自体血清作为冻存保护液。结果:两例患者均获得完全缓解和快速造血功能重建,两例患者均在干细胞回输后的第11天重建造血,无移植相关死亡。结论:采用改良的DICE+G-CSF±R方案动员外周血干细胞动员效果好;用自体血浆代替人血白蛋白作为冻存保护液安全有效,避免了二次污染,值得临床进一步探讨。     

两次输注自体干细胞移植治疗血液肿瘤4例报告

目的:观察两次干细胞输注在自体外周血干细胞移植(APBSCT)中对中性粒细胞及血小板减少的影响,观察其对患者并发感染和出血的影响.方法:对4例血液肿瘤患者经化疗 G-CSF动员并采集外周血干细胞,预处理后分别于0、 3天回输外周血干细胞采集物,观察外周血ANC、BPC及移植相关并发症和血液制品支持治疗情况.结果:用Cy AraC G-CSF动员可获足够CD34 细胞,患者ANC低于0.5×109/L为5～12天,BPC持续低于20×109/L仅为3～4天,仅1例因输血小板发热,仅1例因口腔出血点输血小板1单位,移植费用降低.结论:两次干细胞输注在自体干细胞移植中可缩短粒细胞及血小板减少时间,并减少感染和出血,降低支持治疗费用.     

纯化的自体CD34+细胞移植治疗多发性骨髓瘤的临床观察

目的采用CD34+细胞体外分选技术对多发性骨髓瘤患者进行自体CD34+细胞移植,以降低自体移植后肿瘤复发率.方法对1例多发性骨髓瘤患者采用CliniMACS临床型细胞富集仪,利用磁性分选技术收集CD34+细胞,患者于预处理后,输注分选后的CD34+细胞.结果 CD34+细胞体外纯化富集可使回输CD34+细胞中位数达3.9×106/kg,CD34+细胞回收率为35%,纯度达到97.6%;移植后,患者于+14 d时,中性粒细胞数＞0.5×109/L,+35 d时血小板＞20×109/L,提示造血重建.结论 CD34+细胞移植后获得迅速、稳定的造血重建.体外CD34+细胞纯化富集后移植可望提高多发性骨髓瘤患者自体移植疗效.     

自体骨髓间充质干细胞和外周血造血干细胞共移植治疗恶性淋巴瘤(附1例报告)

目的探讨自体骨髓间充质干细胞(bone marrow-mesenchymal stem cell,BM-MSC)和外周血造血干细胞(PBSC)共移植治疗恶性淋巴瘤(NHL)的安全性和疗效.方法对1例确诊为NHLⅡ期A组的患者,采用CHOP方案(环磷酰胺750 mg/m2、阿霉素40 mg/m2、长春新碱1.4 mg/m2,第1天静滴;强的松60mg/m2,第1～5天口服)化疗4次.自体PBSC动员前取自体骨髓培养BM-MSC,扩增后计数1×107,冻存.动员方案为美罗华、环磷酰胺加粒系集落刺激因子(G-CSF).采集结果单个核细胞(MNC)2.61 ×10s/kg,CD细胞4.27×106/kg,予-80℃低温冻存.CHOP方案预处理后24h先回输自体PBSC,次日回输自体BM-MSC.结果移植后白细胞计数(WBC)和血小板计数(BPC)低谷时间为移植后第6(+6)天和+9天,恢复中性粒细胞绝对计数(ANC)>0.5×109/L和BPC＞20×109/L的时间分别为5天和1天.除口腔溃疡和皮肤感染外,无其他严重的移植相关并发症.结论自体BM-MSC和自体PBSC共移植治疗恶性淋巴瘤安全可行,移植后造血重建快,长期疗效有待进一步积累.     

-80℃简易冻存外周造血干细胞临床应用观察

目的:探索一种简便易行、安全有效、成本低的干细胞的冻存方法。方法:6份外周造血干细胞以1640培养液和二甲基亚砜、10%白蛋白作保护剂在-80℃条件下直接冻存,检测冻存前后外周血干细胞的活性、单个核细胞(MNC)数和CD34+细胞数及回收率。结果:冻存前后外周血干细胞活性、MNC和CD34+细胞的数量及回收率满足移植要求,输注后造血均重建。结论:1640培养液和二甲基亚砜、10%白蛋白作保护剂在-80℃条件下直接冻存干细胞是一种简便、可靠、安全、成本低的外周血干细胞冻存方法。     

自体干细胞移植治疗类风湿性关节炎

目的探讨自体干细胞移植治疗难治性类风湿性关节炎的疗效。方法对1例类风湿性关节炎患者进行自体骨髓移植,预处理方案采用环磷酰胺、ATG。结果移植后造血恢复较快,干细胞回输后第10天白细胞升至1.0×109/L以上,干细胞回输后第5天患者关节疼痛消失,可下床活动。移植后2个月RF、CRP、ESR均恢复正常。结论自体干细胞移植有可能作为类风湿性关节炎的有效方法。     

-80℃冻存外周血造血干细胞的应用研究

目的:探求一种操作简便实用、造血干细胞回收率和存活率高、成本低的外周血造血干细胞的冻存方法。方法:CP-1和5%白蛋白作保护剂在?80℃条件下直接冻存,并检测冻存前后外周血干细胞的活性,单个核细胞(MNC)和CFU-GM、CD34 细胞的回收率。结果:冻存前后外周血干细胞活性,单个核细胞(MNC)和CFU-GM、CD34 细胞的回收率无明显差异(P>0.05),输注后造血均重建。结论:CP-1和5%白蛋白作保护剂在?80℃条件下直接冻存干细胞是一种简便、可靠、安全、成本低的外周血干细胞冻存方法,具有广泛的临床推广价值。     

深低温保存自体外周血干细胞移植治疗多发性骨髓瘤的临床研究

目的探讨自体外周血干细胞移植(APBSCT)治疗多发性骨髓瘤(MM)的临床疗效及安全性。方法1例MM患者接受APBSCT治疗,采用环磷酰胺(CTX) 粒细胞集落刺激因子(G-CSF)方案动员干细胞,CS-3000血细胞分离机采集外周血干细胞并保存于-80℃冰箱;用马法兰200 mg/m2方案预处理后,解冻回输冻存的干细胞。观察患者移植前后的临床疗效。结果动员后获得单个核细胞数(MNC)3.705×108/kg;CD34 细胞数2.56×106/kg。患者获得造血重建,中性粒细胞数(ANC)>0.5×109/L、血小板计数(PLT)>20×109/L的时间分别是12 d、15 d。移植后随访1年患者处于完全缓解(CR)状态。结论APBSCT治疗MM安全有效。     

自体造血干细胞冷冻保存方法的实验研究

目的 观察程序降温仪冷冻法和HES冷冻法在冷冻保护体系和技术实施上的差别，选择一种可以获得良好回收率，操作过程更为简便和安全的冷冻保存方法。方法 (1)通过血细胞分离机，获得去除了红细胞后的单个核白细胞。(2)使用程序降温仪冷冻法，将细胞冷冻后，放置液氮中保存。(3)使用HES冷冻法，直接置-80℃低温冰箱中保存。(4)采用常规计数法，台盼蓝拒染法，甲基纤维素法，流式细胞仪，检测两种方法冻存自体造血干细胞前后，单个核细胞的总数、细胞存活数，CFU—GM集落形成数、CD34阳性细胞的百分率。(5)在37℃恒温水浴槽中，快速解冻细胞。结果 用两种冻存方法保存的自体造血干细胞，解冻回输给患者后，均未出现过敏症状和特殊的不良反应。检测两种不同冻存方法，在冻存前后单个核细胞的总数、细胞存活率、以及CFU—GM集落形成数和CD34阳性细胞的百分率。经统计学处理，无显著性差异。在操作时间上，程序降温仪冷冻法为4h，HES冷冻法为1．5h。结果表明，采用HES冷冻体系，直接置-80℃低温冰箱短期冻存自体造血干细胞，回收率不受影响，方法简单，操作时间短，临床使用安全可靠。     

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.