4℃与-196℃保存胸腺细胞的比较

目的探讨4℃与-196℃保存胸腺细胞的存活率。方法 取大白鼠,无菌条件下制备成胸腺细胞悬液,分别于4℃与-196℃保存,保存36h和35d后,用Trypanblue染色、观察胸腺细胞的回收率。结果 胸腺细胞4℃保存36h,回收率为28％;-196℃冻存35d,细胞回收率为2318／μl。结论 -196℃冻存胸腺细胞其存活率高于4℃保存,-196℃保存胸腺细胞是一种有效的中长期保存方法。     

骨髓细胞低温(4℃)保存的实验研究和临床初步应用

本文通过10只兔和13例人骨髓细胞4℃保存,证明TC199对骨髓细胞4℃保存是有效的。EDTA·Na_2对骨髓细胞的抗凝能力比枸橼酸钠强,肝素抗凝维持时间短,容易产生凝结。TC199-EDTA·Ha_2(TCE)保存液与等量骨髓细胞混悬后,在4℃保存3天,骨髓细胞的台盼蓝和伊红拒染细胞的百分率和有核细胞回收率均在70％以上,骨髓粒系祖细胞(Granulocyte-Macrophage Colony-Forming Unit In Culture,简称CFU-C)的回收率为80％,保存5天时骨髓细胞中的粒细胞将出现固缩等退行性变化。临床资料初步表明,4℃保存3天的TCE骨髓细胞的输注是安全的,未见异常反应,可以加速放疗和化疗后骨髓及血液学的恢复。     

普通冰箱4℃短期保存小鼠骨髓单个核细胞的研究

目的:建立小鼠骨髓单个核细胞短期保存简便有效的方法。方法:采用密度梯度离心法获得小鼠骨髓单个核细胞,DMEM培养液为小鼠骨髓单个核细胞冷藏营养液,直接置骨髓单个核细胞于4℃冰箱中保存,分别在当天至60 h选择性观察细胞形态,测其细胞回收率、活力及细胞周期。结果:小鼠骨髓单个核细胞置4℃冰箱保存2 d,活细胞率达80%以上;细胞回收率达70%以上;细胞周期与对照组相比无明显差别。结论:4℃冰箱能在2 d内保存小鼠骨髓单个核细胞,是一种简便易行的短期保存方法。     

普通冰箱4 ℃短期保存小鼠骨髓单个核细胞的研究

目的：建立小鼠骨髓单个核细胞短期保存简便有效的方法。方法：采用密度梯度离心法获得小鼠骨髓单个核细胞,DMEM培养液为小鼠骨髓单个核细胞冷藏营养液,直接置骨髓单个核细胞于4 ℃冰箱中保存,分别在当天至60 h选择性观察细胞形态,测其细胞回收率、活力及细胞周期。结果：小鼠骨髓单个核细胞置4 ℃冰箱保存2 d,活细胞率达80%以上;细胞回收率达70%以上;细胞周期与对照组相比无明显差别。结论：4 ℃冰箱能在2 d内保存小鼠骨髓单个核细胞,是一种简便易行的短期保存方法。     

家兔睾丸移植模型研究

目的 比较不同睾丸血管吻合方式及睾丸保存方式对移植后睾丸存活率及生精功能的影响。方法 分别将睾丸血管与睾丸血管、腹壁下血管、隐血管吻合，建立器官移植模型；分别用UW液和HTK液保存睾丸，建立器官低温保存移植模型。术后监测睾丸存活率、精子生成及雄激素分泌水平。结果端端吻合睾丸动静脉、输精管的睾丸阴囊原位移植方法最为简单易行，存活率高，术后生精功能良好；在0℃～4℃条件下，UW液和HTK液分别可保存兔睾丸72h和60h。结论 睾丸原位移植最为方便可靠，存活率高，术后生精功能良好；在0℃～4℃条件下，UW液和HTK液保存兔睾丸不宜超过72h和60h。     

Research on short-term preservation of mouse bone mononuclear cell in the ordinary refrigerator at the temperature of 4 ℃

目的:建立小鼠骨髓单个核细胞短期保存简便有效的方法.方法:采用密度梯度离心法获得小鼠骨髓单个核细胞,DMEM培养液为小鼠骨髓单个核细胞冷藏营养液,直接置骨髓单个核细胞于4℃冰箱中保存,分别在当天至60 h选择性观察细胞形态,测其细胞回收率、活力及细胞周期.结果:小鼠骨髓单个核细胞置4℃冰箱保存2d,活细胞率达80％以上；细胞回收率达70％以上；细胞周期与对照组相比无明显差别.结论:4℃冰箱能在2d内保存小鼠骨髓单个核细胞,是一种简便易行的短期保存方法.     

血清放置时间对大鼠血生化检测的影响

目的探讨大鼠血清放置时间对14项血生化检测结果的影响。方法随机选取40只成年SD大鼠,空腹采集静脉血液标本,静置1 h,分离血清并密封分装,存放于4℃和-20℃冰箱内,应用全自动生化分析仪于分离血清后0 h、4 h、24 h、96 h、7 d检测谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、总蛋白(TP)、白蛋白(ALB)、肌酐(CREA-J)、尿酸(UA)、尿素氮(UREA)、血糖(GLU)、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、肌酸激酶(CK)和乳酸脱氢酶(LDH)等14项血生化指标,并进行比较。结果雌雄大鼠血生化数据变化趋势一致。4℃保存时,与0 h相比,ALP在4 h、24 h、96 h、7d均显著降低(P0.05),ALB在96 h和7 d均极显著升高(P0.01),CREA-J在96 h和7 d均显著升高(P0.05),UA在24 h、96 h、7 d均极显著升高(P0.01),其他指标差异无显著性(P0.05)。-20℃保存时,与0 h相比,ALT在7 d显著升高(P0.01),AST在96 h和7 d显著升高(P0.05),TP在4 h和24 h显著降低(P0.05),ALB在4 h、24 h、96 h、7 d均显著升高(P0.01),CREA-J在24 h、96 h、7 d均显著升高(P0.01),UA在4 h、24 h、96 h、7 d均显著升高(P0.01),TC在4 h、24 h、96 h、7 d均显著升高(P0.05),TG在96 h和7 d显著升高(P0.05),CK在96 h和7 d显著升高(P0.05),LDH在96 h和7 d显著升高(P0.05),其他指标差异无显著性(P0.05)。结论大鼠血生化测定尽可能在采集血清后即刻完成,尤其是ALP的测定,4℃冷藏时,测定UA不宜超过4 h,测定ALB和CREA-J不宜超过24 h,测定ALT、AST、TP、UREA、GLU、TC、TG、LDL-C、CK及LDH等指标不宜超过7 d。     

红细胞冷冻干燥保存研究进展

随着临床成分输血技术的推广及成熟,红细胞保存方法的研究倍受关注及重视。目前红细胞的常规保存方法主要有液体保存法（4℃）和深低温保存法（-80℃或-196℃）。液体保存法（4℃）保存时间短,最长只能有效延续至42d,而且容易受到细菌污染;深低温保存（-80℃或-196℃）可长期保存,但需要昂贵、笨重的低温设备,而且由于保护液中含有甘油等渗透性保护剂,解冻后需要反复洗涤。而冷冻干燥保存法以其重量大大减轻、便于运输、适合室温保存、易于再水化等优势备受关注。本文就红细胞保存的相关理论及冷冻干燥法保存红细胞研究的进展及所面临的挑战进行讨论,从而为发展一种安全、简单和有效的红细胞冷冻干燥保存方法提供理论指导。     

4℃冰箱保存骨髓及外周血造血干细胞

目的　为骨髓造血干细胞 (BMSC)及外周血造血干细胞 (PBSC)的短期保存建立简便有效的方法。方法　采用自体血浆为骨髓单个核细胞 (BMMCs)及外周血单个核细胞 (PBMCs)冷藏营养液 ,直接置BMMCs及PBMCs于 4℃冰箱中保存 ,分别在当天至第 7d选择性测其细胞活力 ,CD+ 3 4 细胞阳性率及粒 -单系造血祖细胞 (CFU -GM )集落形成能力。结果　BMMCs及PBMCs置 4℃冰箱保存 3d后 ,台盼兰拒染率、CD+ 3 4 细胞回收率和CFU -GM回收率分别达 80 %以上 ;保存 4d后 ,上述三项指标仍达 70 %以上。结论　当患者接受高剂量化疗时用此方法能在 4d内安全地保存BMSC或PBSC。     

低温下皮肤力学信号相关蛋白表达与活力关系研究

目的:探讨皮肤力学信号相关蛋白整合素(integrin)、踝蛋白(talin)在不同低温条件下的表达与皮肤活力改变的关系,进一步揭示皮肤低温损伤的机制。方法:从同一个体取标本后用5种不同的温度保存,分为新鲜组、4℃组、-20℃组、-80℃组和-196℃组,并在保存48 h后进行实验观察,用integrinβ1、talin抗体对5种储存条件下的皮肤作免疫组化染色,图像分析方法定量,同时采用氧耗量的方法对不同低温保存后皮肤活力进行检测。结果:新鲜组、-196℃组、-80℃组、-20℃组、4℃组活力依次下降,同时-80℃组、-20℃组、4℃组两种蛋白表达亦有明显下降。结论:低温保存后活力的改变与皮肤力学信号相关蛋白的表达下降关系密切。     

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.