慢速冷冻和玻璃化冷冻对人类卵巢组织中卵泡形态的影响

【目的】比较慢速冷冻、麦管玻璃化冷冻和尼龙网玻璃化冷冻对人类卵巢组织中卵泡形态的影响。【方法】16例人卵巢组织切成薄片随机分配到新鲜卵巢组（A组1、麦管玻璃化冷冻组（B组）、尼龙网玻璃化冷冻组（C组）和慢速冷冻组（D组）后行组织学和电镜检查。【结果】A、B、C、D组中形态正常的原始卵泡比例分别为72．5％±8．4％、65．6％±12．8％、66．1％±11．1％、48．4％±13．3％；形态正常的初级卵泡比例分别为62．0％±13．9％、58．1％±7．9％、59．0％±16．2％、37．0％±14．0％。D组中形态正常的原始卵泡比例和初级卵泡比例均明显低于A、B、C组。B、C组形态正常的原始卵泡比例和初级卵泡比例。与A组相比无统计学差异。B、C组中形态正常的原始卵泡超微结构无明显改变，D组中形态正常的原始卵泡超微结构存在一定程度的改变。【结论】玻璃化冷冻是人类卵巢组织较适宜的冷冻保存方法。     

玻璃化冷冻对人卵巢组织形态结构及组织增殖活性的影响

目的:探讨玻璃化冷冻对人类卵巢组织卵泡形态、超微结构和组织增殖活性的影响。方法:采用12例人卵巢组织切成薄片随机分配到新鲜卵巢组(A组)和玻璃化冷冻组(B组),两组行组织学光镜和透射电镜检查卵泡形态及结构变化,免疫组化测定卵巢组织中增殖细胞核抗原(PCNA)的表达情况。结果:A、B两组中原始卵泡和初级卵泡分布比例分别为86.4%、13.6%和84.5%%、15.5%,两组比较差异无统计学意义(P>0.05)。光镜下A、B两组中形态正常的原始卵泡分别为75.3%和69.3%,两组比较差异亦无统计学意义(P>0.05);但B组冻融后初级卵泡形态异常的比例(56%)高于A组(28.2%),两组比较差异有统计学意义(P<0.05);A、B组中形态正常的原始卵泡超微结构无明显改变,B组中初级卵泡超微结构存在一定程度的改变。A、B两组卵巢中PCNA阳性表达比较差异无统计学意义(P>0.05),PCNA阳性表达主要见于原始和初级卵泡的卵母细胞、颗粒细胞和卵巢组织间质细胞。结论:玻璃化冷冻是一种较适宜保存人类卵巢组织的方法。     

不同玻璃化冷冻载体对人卵巢组织冷冻效果的影响

目的: 比较不同的玻璃化冷冻载体对人卵巢组织冷冻效果的影响,为人卵巢组织玻璃化冷冻方案的选择提供理论基础和实验参考依据.方法: 预处理人卵巢皮质切成10 mm×1 mm×1 mm大小,随机分为新鲜组(A组)、针浸润玻璃化冷冻组(B组)、表面玻璃化冷冻组(C组)和直接覆盖玻璃化冷冻组(D组).A组直接予以固定、组织切片及HE染色;B、C、D 3组采用不同玻璃化冷冻载体冷冻后,投入液氮保存2个月,采用快速复温法解冻后予以固定、切片及染色.行形态学分析,通过计算各组卵巢组织切片中正常形态始基卵泡率,比较各组冷冻效果.结果: 4组共计数809个始基卵泡,其中正常形态始基卵泡率分别为:A组96.12%,B组88.14%,C 组80.00%,D组81.25%.行玻璃化冷冻的3组卵巢组织正常形态始基卵泡率均明显小于A组(P< 0.01);而B组、C组和D组正常形态始基卵泡率差异均无统计学意义(P >0.05).结论: 玻璃化冷冻后人卵巢组织正常形态始基卵泡率较新鲜组织下降,但仍能较好地保存人类卵巢组织,可用于人类卵巢组织的冷冻保存.     

玻璃化冷冻人卵巢组织形态学研究

目的：探讨玻璃化冷冻及体外培养对人卵巢组织内各级卵泡的形态影响。方法采用液氮直接覆盖玻璃化冷冻（DCV ）方法冷冻保存成人卵巢组织,复苏后行体外培养,观察卵巢组织中卵泡的形态学改变。结果新鲜培养组与新鲜组、冷冻培养组与冷冻组相比,原始卵泡所占比例明显下降,初级卵泡和次级卵泡构成比显著增加（P＜0．05）;新鲜培养组较新鲜组形态正常的原始、初级、次级卵泡比例降低（P＜0．01）;冷冻组形态正常的初级、次级卵泡比例均低于新鲜培养组（P＜0．05）;冷冻培养组形态正常的次级卵泡比例低于新鲜培养组,差异有统计学意义（P＜0．05）。结论 DCV方案卵泡形态结构有一定损伤,体外培养有一定修复作用。     

人类卵巢组织玻璃化冷冻及复苏后形态学及组织增殖活性观察

目的 探讨玻璃化冷冻对成人卵巢组织中原始卵泡与初级卵泡的形态学及细胞增殖状态的影响.方法 采用液氮直接覆盖玻璃化冷冻(direct cover vitrification,DCV)方法冻存成人卵巢组织块,液氮保存2周后复苏组织.观察和比较冻融前后人卵巢组织中原始卵泡与初级卵泡组织形态学改变;免疫组化染色测定经体外培养48 h后新鲜和冻融卵巢组织中增殖细胞核抗原(PCNA)的表达情况.结果 冻融前后人卵巢组织内原始卵泡和初级卵泡分布差异无统计学意义(P>0.05);新鲜卵巢组织中,形态异常的原始卵泡比例与初级卵泡比例差异无统计学意义(P>0.05),冻融卵巢组织中,形态异常的原始卵泡比例低于形态异常的初级卵泡比例(P<0.05);新鲜组、新鲜及冻融后培养组均有PCNA的阳性表达,PCNA 阳性表达可见于中间型卵泡和初级卵泡的卵母细胞和颗粒细胞、卵巢组织间质细胞.结论 利用DCV方法对人卵巢组织进行冻存能够保存原始卵泡和初级卵泡,冻融过程对原始卵泡和初级卵泡形态结构都可能造成一定程度的损伤,冻融后组织内卵泡有体外生长发育的潜力.     

采用不同载体的玻璃化冷冻对人卵巢组织冻存效果的比较研究

目的比较采用不同载体的玻璃化冷冻方案对人卵巢组织的卵泡形态、冻融后体外培养时卵泡生长和激素分泌能力的影响,寻找临床上简便易行且保存效果上佳的人卵巢组织玻璃化冷冻方案。方法将16例患者的卵巢组织切割成皮质小条后随机分配到新鲜组和不同载体玻璃化组,包括冷冻管组、最小微滴组、不锈钢网筛组和固相表面玻璃化组。观察各组卵巢组织卵泡形态;冻融后的卵巢组织行体外培养,比较卵泡生长情况以及培养液中雌二醇和孕酮的浓度。结果冻融后各组原始卵泡正常形态率均低于新鲜组,差异有统计学意义（P〈0.05）。冷冻管组、最小微滴组、不锈钢网筛组和固相表面组,原始卵泡形态正常率分别为（64.57±11.84）%、（78.36±7.62）%、（77.21±6.51）%和（84.70±5.03）%。固相表面组原始卵泡形态正常率明显高于其他玻璃化组（P〈0.05）。体外培养液中雌二醇和孕酮的平均水平随培养时间逐渐升高。固相表面玻璃化组两种激素平均浓度明显高于其他玻璃化组。培养后卵巢组织中原始卵泡所占比例均较新鲜组下降,生长卵泡比例升高。固相表面玻璃化组培养后生长卵泡比例明显高于其他玻璃化组。结论固相表面玻璃化冷冻可以保存大部分原始卵泡的正常形态,并能较好地保存其体外生长和性激素分泌功能。该技术简便易行且冷冻效果好,适合人卵巢组织玻璃化冷冻的临床使用。     

应用自研载体的玻璃化冷冻与慢速程序化冷冻保存羊卵巢组织的效果比较

目的 自研冷冻载体用于羊卵巢组织的玻璃化冷冻,探讨自研载体对羊卵巢组织的冷冻保存效果。 方法 将获取的32只羊卵巢通过简单随机抽样方式分配到新鲜组、程序化冷冻组、自研载体I玻璃化冷冻组及自研载体Ⅱ玻璃化冷冻组,观察各组卵巢组织形态、增殖与凋亡及培养后雌激素分泌水平。结果 冷冻复苏后程序化冷冻组、自研载体I玻璃化冷冻组及自研载体Ⅱ玻璃化冷冻组原始卵泡形态正常率分别为74.2%、72.8%、72.3%,低于新鲜组83.7%(χ²=13.079,P=0.004),程序化冷冻组初级卵泡正常形态率低于新鲜组(χ²=12.486,P=0.000),2个玻璃化冷冻组初级卵泡正常形态率与新鲜组比较差异无统计学意义(P=1.000,P=0.972);各组之间激素水平及增殖细胞核抗原阳性表达率差异无统计学意义(F=0.363,P=0.780;χ²=0.359,P=0.949);冷冻复苏各组细胞凋亡数目均明显高于新鲜组(F=37.584,P=0.000),程序化冷冻组细胞凋亡数目明显高于自研载体I玻璃化冷冻组和自研载体Ⅱ玻璃化冷冻组(F=18.992,P=0.000)。结论 应用自研载体的玻璃化冷冻与慢速程序化冷冻相比能更好地保存大块羊卵巢组织的细胞活性。     

BMP-4对人卵巢组织玻璃化冷冻卵泡作用的实验研究

目的 将人卵巢组织进行玻璃化冷冻和冷冻后体外培养,在培养中加入骨形态发生蛋白-4（BMP-4）,探讨其在玻璃化冷冻后人卵泡的生长、转化和存活方面的作用。方法 32例人卵巢组织标本分为新鲜组,玻璃化冷冻复温后卵巢组织随机分为未培养组、基本培养液组和BMP-4培养液组,前两组直接行组织学检查,后两组体外培养15d后行组织学检查、雌孕激素测定,评估卵泡存活和生长发育情况。结果 玻璃化冷冻后未培养组与新鲜组比较,原始卵泡、生长期卵泡占总卵泡的比率及形态正常的原始和初级卵泡百分率均无显著性(P>0.05);培养结束后基本培养液组和BMP-4组原始卵泡比例显著降低,生长卵泡比例显著升高,BMP-4组孕激素浓度显著低于基本培养液组(P<0.01)。结论 BMP-4有助于冷冻后人卵巢组织体外培养中原始卵泡向初级卵泡的转化,可降低体外培养中颗粒细胞孕激素的基础分泌,有利于卵泡体外存活,可能是玻璃化冷冻卵泡生长发育过程中的促进因子。 [关键词 ] 卵泡;玻璃化冷冻;体外培养;骨形态发生蛋白-4;     

豚鼠卵巢组织片和分离的窦前卵泡玻璃化冷冻效果比较

目的：比较分析不同玻璃化冷冻配方对豚鼠卵巢组织片和单个窦前卵泡形态、存活率及体外发育能力的影响。方法：取5周龄雌性豚鼠卵巢,实验分新鲜对照组、组织片A液毒性组、组织片B液毒性组、卵泡A液毒性组和卵泡B液毒性组,及相应的冻融组,即组织片A液冻融组、组织片B液冻融组、卵泡A液冻融组、卵泡B液冻融组,共9组行毒性试验和冷冻保存。各组卵巢组织片行HE染色形态学观察,分离窦前卵泡经锥虫蓝染色活力测试并行体外培养。结果：A液冻融组和B液冻融组豚鼠卵巢组织片HE染色光镜观察可见大部分卵泡形态正常,A液组和B液组异常窦前卵泡数与新鲜对照组比较差异有统计学意义（P〈0．05）;在分离的窦前卵泡锥虫蓝染色活力测试中,冻融卵巢组织片A组的卵泡复苏率（为66．1％）显著低于（P〈0．05）冻融卵泡B液组（为78．5％）和新鲜对照组（为87．5％）;其余组别之间差异无统计学意义。体外培养卵泡发育情况组织片冷冻组比卵泡冷冻组差,以A液组尤甚,第6天存活卵泡数目不足（为20．0％）,与新鲜对照组（为60．0％）及卵泡B液冻融组（为50．0％）相比较差异有统计学意义（P〈0．05）。结论：玻璃化冷冻法能较好地冷冻保存豚鼠卵巢组织片和分离的窦前卵泡,而单个豚鼠窦前卵泡玻璃化冷冻优于卵巢组织片的玻璃化冷冻;B液较A液更适合豚鼠卵巢卵泡的玻璃化冷冻保存。窦前卵泡的体外培养证明玻璃化冷冻复苏卵泡具有一定的继续发育能力。     

人卵巢组织两种超低温冻存方法的研究

【目的】探索两种人卵巢组织冷冻方案用于生殖细胞保存的效果。【方法】活检卵巢组织来自15名经知情同意的手术患者。组织被随机分配到新鲜组、玻璃化冷冻组和慢速冷冻组。观察和比较各组卵巢组织冻融后的原始卵泡形态完整率,并通过体外培养系统测定培养液中的雌二醇和孕酮水平,观察和比较冻融后卵巢组织内分泌功能。【结果】本研究中新鲜组、慢速冻融组和玻璃化冻融组的原始卵泡形态完整率分别是97.6%、72.6%和80.3%。两冻融组与新鲜组相比明显下降(P<0.001),而在两冻融组间差异并无统计学意义(P=0.237)。在体外组织培养期间,两种冻融卵巢组织都能持续分泌雌二醇和孕酮,两组间差异无统计学意义(P>0.05)。【结论】本研究中两种人类卵巢组织超低温冷存的方法是可行的,可适应不同需要。     

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.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.     

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.