鼠成熟卵母细胞冷冻与复苏的实验研究

目的 探讨不同的植冰温度和冷冻保护剂中蔗糖浓度对小鼠MⅡ期卵子冷冻复苏后形态结构及发育潜能的影响.方法 共收集小鼠MⅡ期卵子372个,随机分为A、B、C三组冷冻.A组冷冻保护液含0.1 mol/L蔗糖,在-6℃植冰;B组冷冻保护液中蔗糖浓度同A组,在-7℃植冰;C组冷冻保护剂中蔗糖浓度为0.2 mol/L,在-7℃植冰.分别对冷冻复苏后的卵子进行质量评估、受精及胚胎培养,观察并比较其受精及卵裂情况.结果 MⅡ期卵子复苏后存活率以及优质卵率A组较B组好,但差异无统计学意义(P>0.05);A组卵子冻融后的受精卵卵裂率(82.35%)高于B组(38.46%),其差异有统计学意义(P<0.05);而受精率和优质胚胎率虽然A组高于B组,但无统计学差异(P>0.05).C组的卵子冻融后存活率(82.61%)、优质卵率(78.26%)、受精卵卵裂率(76.92%)均分别高于B组(68.24%,52.94%,38.46%),其差异有统计学意义(P<0.05);受精率和优质胚胎率C组与B组相比较无统计学差异(P>0.05). 结论 植冰温度升高不影响MⅡ期卵子的冻融效果,而且可能有助于保护卵子的发育潜能;在冷冻保护液中,0.2mol/L蔗糖浓度比0.1mol/L更适合于MⅡ期卵子的冷冻.     

小鼠卵母细胞程序化和玻璃化冷冻效果的比较

目的 本实验通过建立小鼠的实验模型,比较程序化和OPS玻璃化冷冻效果,探讨卵子冷冻的具体方法 ,为卵子冷冻技术应用于临床提供实验依据.方法 通过促排卵获取小鼠MII期卵母细胞,随机分组,观察不同冷冻方法 、高浓度的冷冻保护液,对卵子的成活及进一步发育潜能的影响;不同的IVF前培养时间对冻融卵子的受精及继续发育的影响.结果 (1)OPS玻璃化冷冻组与程序化冷冻组的存活率、受精率、卵裂率及6-8cell胚形成率均无显著差异.(2)玻璃化冷冻组与暴露组的卵母细胞存活率分别为35.3%,100%,有显著性差异(P＜0.05).暴露组与对照组比较的受精率、卵裂率,无显著性差异(P＞0.05).而暴露组与对照组的6-8cell的胚形成率分别为41%,63%,两组比较,有显著差异(P＜0.05).(3)无论OPS玻璃化冷冻法还是程序化冷冻法,冻融后的培养2 h组和培养1 h组的,受精率、卵裂率,两组比较,均有显著性差异(P＜0.05).结论 本实验证实,OPS玻璃化冷冻法至少可以达到程序化冷冻法同样的效果,而OPS玻璃化冷冻法有好于程序化冷冻法的趋势.加之,其简便、省时、经济等优点,因而,更有发展前景.     

玻璃化冷冻法对冻融胚胎移植周期结局的影响

目的:探讨玻璃化冷冻法对冻融胚胎移植周期结局的影响.方法:对324个玻璃化冷冻人卵裂期胚胎复苏周期和146个程序化慢速冷冻胚胎复苏周期进行分析,比较2组冷冻胚胎复苏后的存活率、完整率、种植率、临床妊娠率、孕周、新生儿身高及体质量等指标.结果:玻璃化冷冻组的胚胎完整率、临床妊娠率均高于程序化慢速冷冻组(83.24％ vs...     

玻璃化冷冻在卵巢过度刺激综合征全胚胎冷冻中的应用

目的比较慢速程序化冷冻法(慢速法)和玻璃化冷冻法(玻璃化法)对卵巢过度刺激综合征全胚胎冷冻分裂期胚胎发育潜能的影响。方法选择为避免过度刺激而行全胚胎冷冻的不孕患者162例,按其冷冻方法的不同,分成慢速程序化冷冻组和玻璃化冷冻组,并对其随后进行首次解冻移植的胚胎复苏率、胚胎种植率、妊娠率和多胎妊娠率进行分析。结果玻璃化冷冻组72例胚胎解冻后,其溶解率、复苏率、种植率、妊娠率、多胎妊娠率分别为7.84%、96.08%、52.38%、69.44%和50.00%,慢速程序化冷冻组90例胚胎解冻后,上述指标分别为17.79%、90.38%、30.32%、52.22%和19.15%,两组差异有统计学意义(P<0.05)。结论玻璃化冷冻法能够更好地保存胚胎复苏后的发育潜能,更适合卵巢过度刺激综合征患者胚胎的冷冻保存。     

两种胚胎冻融方法对累积妊娠率的影响

目的比较玻璃化法和程序法对人第3天卵裂期胚胎冻融移植周期累积妊娠率的影响。方法选取程序化冻融周期92例、玻璃化法冻融周期179例,比较分析这两种冻融方法在存活率、累积临床妊娠率等方面的效果。结果玻璃化法在胚胎复苏存活率(94.3%vs53.9%,P<0.01)、复苏完整胚胎形态(89.4%vs36.3%,P<0.01)、累积临床妊娠率(28.2%vs43%,0.010.05)。结论与程序化法比较,玻璃化法可以提高累积妊娠率,是一种高效胚胎冻融方法 。     

两组冷冻保护剂对卵裂期胚胎玻璃化冷冻效果的研究

目的：评价两组玻璃化冷冻保护剂对卵裂期胚胎的冷冻效果。方法以体外受精-胚胎移植周期患者受精后第3天废弃胚胎为实验对象,共102枚胚胎入选,随机分为乙二醇＋丙二醇（EG＋PROH）组52枚,乙二醇＋二甲基亚砜（EG＋DMSO）组50枚。比较观察两组玻璃化冷冻保护剂对卵裂期胚胎冷冻解冻后胚胎的复苏效果。结果玻璃化冻融后两组在复苏率上无显著差异,但在完整胚胎复苏率和卵裂球复苏率上EG＋PROH组显著高于EG＋DMSO组（P＜0.05）。结论对于卵裂期胚胎的玻璃化冷冻,选用EG＋PROH作为冷冻保护剂可取得较好的效果。     

不同冷冻方法对小鼠卵母细胞冻存的影响

目的：：探讨不同冷冻方法对小鼠卵母细胞存活率及其发育潜能的影响。方法：采用玻璃化冷冻技术,选取开放式载体与封闭式载体分别冻存带卵丘细胞(COC)或剥除卵丘细胞(OD)的小鼠卵丘复合体,复苏后分别作体外受精,胚胎培养。结果：开放式载体冻存组存活卵数76.79%略高于封闭式载体冻存组72.98%,无统计学意义(P >0.05);两组内 COC 组的卵子存活率显著高于 DO 组,但有统计学差异(P <0.05),COC 组的受精率、囊胚率略高于 DO 组,无统计学差异(P >0.05)。结论：(1)封闭式载体有效避免了潜在的污染问题,但不同载体对卵子存活率无显著影响。(2)卵丘细胞在玻璃化冷冻卵母细胞中可以起到保护作用,并对胚胎后期的发育有一定的改善。     

D3卵裂期胚胎玻璃化冷冻技术在IVF实验室中的应用

目的探讨玻璃化冷冻技术在人类卵裂期胚胎冻存中的应用价值。方法回顾性分析本中心261个冷冻胚胎复苏周期,比较2种冷冻方法的胚胎存活率、完好率、妊娠率和种植率等数据。结果玻璃化冷冻组的存活率(97.11%)、完好率(89.02%)和种植率(23.12%)显著高于程序化组(91.31%、63.73%、16.31%)(P<0.05),而妊娠率和平均年龄、平均移植胚胎数、平均内膜厚度两组间无统计学差异(40.26%vs40.22%、31.3±5.3vs32.2±5.5、2.25±0.55vs2.27±0.70、8.9±1.1vs9.1±1.6)(P>0.05)。结论玻璃化冷冻是卵裂期胚胎保存的优选方法。     

玻璃化冷冻囊胚复苏移植与玻璃化冷冻胚胎复苏后继续培养至囊胚移植的妊娠结局比较

目的比较玻璃化冷冻囊胚复苏后移植与玻璃化冷冻胚胎复苏后继续培养至囊胚移植的临床妊娠结局。方法回顾性分析2013年1月至2014年12月在郑州大学第二附属医院生殖中心行玻璃化冷冻复苏后移植囊胚的不孕症患者134例(134个周期)的妊娠情况,其中卵裂期胚胎培养至囊胚冷冻复苏后移植者79个周期(A组),卵裂期胚胎冷冻复苏后继续培养至囊胚再移植者55个周期(B组)。比较2组患者临床妊娠率、着床率及早期流产率。结果 A组79个周期,复苏囊胚数151个,存活囊胚数133个,复苏率88.08%(133/151);获得可移植囊胚72个周期,妊娠周期46个,妊娠率为63.89%(46/72),着床率51.56%(66/128),早期流产率为15.21%(7/46)。B组55个周期227个胚胎,复苏存活170个胚胎,复苏率74.89%(170/227);其中45个周期共形成可移植囊胚93个,囊胚形成率为54.70%(93/170);获得妊娠周期23个,妊娠率为51.11%(23/45),着床率为36.56%(34/93),早期流产率13.04%(3/23)。A组囊胚复苏率高于B组卵裂期胚胎复苏率(P<0.05),且A组着床率显著高于B组(P<0.05),但2组临床妊娠率、流产率、异位妊娠率比较差异均无统计学意义(P>0.05)。结论玻璃化冷冻囊胚复苏移植能获得较高的着床率和妊娠率,囊胚可能是玻璃化冷冻胚胎的较好阶段。     

两种玻璃化胚胎冷冻方法对不同品系小鼠胚胎冷冻结果比较

目的 探讨EFS和DAP两种玻璃化冷冻方法对不同品系小鼠胚胎冷冻的效果。方法6个品系小鼠（KM、ICR、BALB/c、C57BL/6J、OB/OB、LAP/rtTA0F59）的2-cell胚胎分别用EFS和DAP两种玻璃化冷冻方法进行冷冻和复苏,比较两种冷冻方法的胚胎复苏率和着床率。结果 6个品系小鼠冷冻胚胎EFS方法的平均复苏率为69.97%（47.9%～83.6%）,DAP方法的平均复苏率47.23%（26.3%～76.7%）EFS方法明显优于DAP方法。其中KM、ICR和BALB/c小鼠EFS方法的冷冻复苏率显著高于DAP方法（P<0.01）;冻融胚胎移植后EFS方法的平均着床率27.23%（1.75%～45.0%）,DAP方法的平均着床率31.43%（7.0%～46.3%）。除KM、ICR小鼠外,其他4个品系小鼠的着床率DAP方法高于EFS方法。结论 KM和ICR远交群小鼠胚胎适合用EFS方法冷冻保存;C57BL/6J、OB/OB、LAP/rtTA0F59三个品系小鼠DAP方法优于EFS方法,但差异不大; BALB/c小鼠两种玻璃化冷冻方法的冻融胚胎着床率均较低,需进一步研究。     

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.