人类胚胎冻存技术的初步研究

研究了不同冷冻与复芳方法以及不同移植周期法对人类胚胎冻存结果的影响。共复１４９个胚胎,８９个（５９．７％）得以存活。在２０个移植周期中,５例（２５．０％）获得临床妊娠,２例已经分健康的婴儿。结果表明,采用缓慢降温至－１２０℃再浸入液氮中冷冻及快速升温、逐步置换冷冻保护剂的方法复苏胚胎的存活率（７２．７％）较高,但不同移植周期对冻胚移植的临床妊娠情况影响不大。     

人类胚胎冷冻复苏影响因素分析

目的：观察患者年龄、促排卵方案、受精方式、胚龄及冻胚移植周期方案对于冷冻胚胎复苏和移植后临床妊娠的影响。方法：慢速冷冻体外受精周期中剩余的质量良好的胚胎，随后于不同周期用快速融解法复苏后进行移植。根据上述因素将患者分组，对复苏后胚胎质量和临床妊娠率进行比较。结果：126例患者中，融解胚胎总数445个，复苏后成活368个(82．7％)。获得临床妊娠48例(38．1％)。年龄对冷冻胚胎复苏率和移植后临床妊娠率的影响比较明显。随着年龄增加，冻胚移植的妊娠率下降，尤其是在35岁以后，下降更为明显。促排卵方案、受精方式、冷冻时胚胎的培养天数和移植周期方案均对冷冻胚胎复苏率和移植后临床妊娠率没有明显影响。结论：在诸多因素中，只有患者年龄会影响冻胚移植的结果。     

影响冻融胚胎复苏率和移植成功率因素分析

目的探讨影响冻融胚胎复苏率和移植成功率的相关因素。方法回顾性分析573个冷冻胚胎移植（FET）周期的临床资料,按冷冻前胚胎质量、胚胎发育时间、解冻后胚胎复苏程度和冷冻方法分别进行分组,分析不同分类各组间的胚胎复苏情况和（或）临床妊娠率差异。结果冷冻当天胚胎中有优质胚胎者的临床妊娠率显著高于无优质胚胎者（31．8％vs20．0％）（P〈0．05）。受精第2天冷冻者与受精第3天冷冻者的胚胎复苏率和临床妊娠率差异无统计学意义（79．1％vs82．9％和25．5％vs31．2％）（P〉0．05）。解冻后完全移植100％卵裂球完整者和混合移植者的临床妊娠率显著高于完全移植部分卵裂球损伤者（36．7％vs24．1％和29．2％vs24．1％）（P〈0．05）。冷冻方法改进后的胚胎复苏率和全胚复苏率显著高于冷冻方法改进前（82．0％vs66．3％和50．0％vs27．5％）（P〈0．05）。结论胚胎质量与胚胎复苏率和临床妊娠率密切相关,冷冻胚胎解冻后全胚复苏率与临床妊娠率正相关,冷冻过程中的操作细节是胚胎实验室保证冻融胚胎复苏、移植成功的重要因素。     

卵子冷冻技术的临床应用

目的 探讨卵母细胞冷冻技术在临床应用.方法 将258例接受体外受精-胚胎移植(IVF-ET)和单精子卵浆内注射(ICSI)治疗,取卵数≥20枚的258个周期,分为部分卵母细胞冷冻组(简称冻卵组)84个周期和胚胎冷冻组(简称冻胚组)174个周期,比较两组新鲜周期受精率和临床妊娠率.总结了23个卵母细胞冷冻复苏周期及19个移植周期(其中4个周期为捐赠周期)的复苏率,受精率,卵裂率,胚胎种植率,临床妊娠率,比较冻胚组冻胚复苏移植的临床结局.结果 冻卵组和冻胚组新鲜周期的受精率和临床妊娠率差异无统计学意义(P>0.05);19个卵母细胞冷冻复苏的移植周期与56个冷冻胚胎复苏的移植周期的胚胎种植率和临床妊娠率差异亦无统计学意义(P>0.05);卵母细胞冷冻复苏的移植周期妊娠率为47.4%(9/19),其中冻卵捐赠4个移植周期,2例临床妊娠(1例已分娩).结论 对于接受IVF/ICSI治疗的不孕妇女,由卵巢刺激产生的较多卵母细胞,无论是否选择冷冻部分卵母细胞,对其新鲜周期的受精率和妊娠率没有影响;冻卵移植周期与冻胚移植周期的妊娠率差异也无统计学意义;但是卵母细胞冷冻在生育力保存和分享捐赠方面较冻胚具有显著的优势.     

两种冷冻载体对人胚胎玻璃化冷冻效果的影响

目的探讨不同载体对人早期胚胎玻璃化冷冻复苏的效果。方法采用半封闭式载体冷冻复苏胚胎的患者共265个周期,采用封闭式载体冷冻复苏胚胎的患者共136个周期,均采用玻璃化冷冻办法冷冻体外精-胚胎移植第3日的可利用胚胎,观察胚胎复苏和临床妊娠情况。结果半封闭式麦管冷冻复苏胚胎的患者移植周期258个,取消周期7个;复苏率、临床妊娠率和种植率分别为96．88％、48．44％和32．20％。封闭式麦管冷冻复苏胚胎的患者移植周期132个,取消周期4个;复苏率、临床妊娠率和种植率分别为95．31％、47．72％和32．28％。、两组患者的平均年龄、不孕年限、不孕类型、不孕原因、体外受精或卵胞质内单精子注射（IVF／ICSI）构成比以及胚胎复苏率、移植胚胎数、临床妊娠率和种植率比较差异均无统计学意义（P〉0．05）。结论 封闭式冷冻载体能获得与半封闭式载体同样的冷冻效果,同时能避免胚胎受病原微生物的交叉污染,是冷冻人早期胚胎的理想方法。     

授精第三天4细胞及5细胞期胚胎的冷冻价值

目的 分析授精第3天为4细胞及5细胞期胚胎冻融及移植后对临床结局的影响,探讨其冷冻价值。方法 收集270个冻融胚胎移植（FET）周期,授精第3天胚胎采用程序化冷冻法冻存,快速解冻法复苏后移植,根据冷冻日胚胎所含细胞数比较各细胞数胚胎复苏存活率,并分为4、5细胞组与≥6细胞组,比较两组间的临床妊娠率、种植率及因胚胎因素取消周期率（简称周期取消率）。结果 （1）270个周期共解冻胚胎939枚,复苏存活率为65.3% (613/939);移植胚胎561枚,临床妊娠率27.2%(67/246),种植率15.2%（85/561）;（2）4、5细胞组周期取消率显著高于≥6细胞组（P＜0.01）,且移植胚胎数显著低于≥6细胞组（P＜0.05）,但胚胎复苏成功并移植后两组妊娠率、种植率及活产率均无显著性差异（P＞0.05）。结论 冻存4、5细胞期胚胎与≥6细胞期胚胎妊娠率无统计学差异,提示具有冷冻价值。     

冷融胚胎移植的临床应用(附36例分析)

目的对３６个冷冻胚胎移植周期的治疗做一临床分析。方法用药方案包括自然周期、激素替代周期间（ＨＲＴ）和克罗米芬或人绝短期促性腺激素（ＨＭＧ）刺激周期，第２天胚胎于出现尿黄体生成素（ＬＨ）峰或给予黄体酮的第３－４天移植，第３天胚胎于出现尿ＬＨ峰或给予黄体酮的第４～５天移植。结果　３６例患者进行了３６个周期的冷冻胚胎移植，每周期移植胚胎数（３．５±１．４）个，３６个移植周期中１０次临床妊娠（２７．８％），３例双胎妊娠（ １例于孕中期自动减为单胎）， ７例单胎． ２例妊娠早期自然流产．有 ８例继续妊娠达 ７－３５周。原核期冻融 ５个周期，获临床妊娠 ３次，胚胎期冻融３１个周期，获临床妊娠７次，移植第２天胚胎：于出现尿ＬＨ峰后第４天移植的１０个周期中，１次妊娠：第３天移植的１个周期获妊娠；第２天胚胎于出现尿ＬＨ峰后第４天移植的１６个周期中，５次妊娠；第５天移植的１个周期中，无妊娠。于用黄体酮第４天移植的１周期中无妊娠，第５天移植的３个周期中２次妊娠。结论冻融胚胎移植能获得满意的妊娠率。第２天冻融胚胎的最佳移植时机为出现尿ＬＨ峰的第３大，第３天冻融胚胎为出现尿ＬＨ峰的第４天或于激素替代治疗（ＨＲＴ）周期用黄体酮第５天。     

应用颗粒细胞部分剥除法提高冻融胚胎移植后妊娠率和植入率的临床研究

目的探讨应用颗粒细胞部分剥除法对常规体外受精-胚胎移植（IVF-ET）冻融胚胎移植后妊娠率和植入率的影响。方法140个冻融周期共完成122例移植。实验组70个冻融周期共374个胚胎冷冻前采用颗粒细胞部分剥除法培养，对照组70个冻融周期共392个胚胎冷冻前颗粒细胞完全剥除后培养。复苏后胚胎经激光辅助孵化，培养0．5～3h后移植。比较两组冻融胚胎移植后的妊娠率和植入率。结果对照组完成60个移植周期，临床妊娠率20．00％。植入率7．87％；实验组完成62个移植周期，临床妊娠率37．10％，植入率14．77％，均高于对照组（P〈0．05）。结论应用颗粒细胞部分剥除法可以提高IVF-ET冻融胚胎移植后妊娠率和植入率。     

224例冻融胚胎移植周期分析

目的：探讨胚胎冷冻时机对复苏后胚胎发育的可能影响，并分析移植胚胎数量和质量与冻融胚胎移植妊娠结果的关系。方法：（1）将224个冻融胚胎移植周期，根据冷冻时间分为第2天（D2组）97例，第3天（D3组）127例两组，回顾比较解冻后两组的胚胎存活率、胚胎完好率和临床妊娠率等.（2）比较了移植不同胚胎个数（3、2、1个）和优质胚胎个数（3、2、1、0个）的临床妊娠率。结果：（1）D2组胚胎完好率为54．28％，显著高于D3组的40．65％（P〈0．05），D2组的胚胎存活率、妊娠率、种植率、多胎率和流产率分别为82．57％、26．80％、13．01％、34．62％、11．54％，与D3组（82．17％、37．01％、17．66％、36．17％、17．02％）的差异均无显著性（P〉0．05）。（2）移植3、2、1个胚胎分别可获得34．62％、26．47％、12．5％的临床妊娠率，各组之间比较差异无显著性（P〉0．05）；移植3个或2个优质胚胎的临床妊娠率没有差异，但其临床妊娠率显著高于只移植1个或没有优质胚胎组。结论：胚胎冷冻时机对早期分裂胚胎的发育潜能没有影响。冻融胚胎移植周期的妊娠率与移植的优质胚胎数目有关。     

胚胎复苏后体外孵育时间对妊娠结局的影响

目的探讨复苏后的体外孵育时间对冻存胚胎妊娠结局的影响。方法对100例接受冷冻胚胎移植患者的临床资料进行回顾性分析。结果100例冷冻胚胎移植周期,临床妊娠率为21%,复苏后体外孵育时间的不同妊娠率无显著差异,但流产率差异显著。结论复苏后延长体外孵育时间可以更好的观察胚胎的发育潜能,但增加流产率,应缩短复苏后体外孵育时间。     

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.