小鼠卵母细胞的孤雌激活

目的 探讨一种有效的孤雌激活方法。方法 用乙醇、Ca^2＋载体A23187分别联合6-二甲氨基嘌呤对小鼠卵母细胞进行孤雌激活。结果 7％乙醇卵母细胞活化率、囊胚形成率分别达80.88％、10.29％、;5μmol/L Ca^2＋A23187卵母细胞活率、囊胚形成率分别达86.88％、11.48％。两者差异无显著性。结论 应用乙醇、Ca^2＋载体A23187联合6-二甲氨基嘌呤处理小鼠卵母细胞,均可实现小鼠卵母细胞孤雌发育。     

不同激活方法对小鼠卵母细胞孤雌激活的实验观察

目的　建立合适的小鼠卵母细胞孤雌激活方法。方法　取不同卵龄小鼠卵母细胞 ,运用不同浓度的氯化锶和不同强度的电脉冲对其进行活化 ,观察小鼠卵母细胞激活率和体外发育状况。结果　① 14、16和 18h卵龄组卵母细胞经 10mmol LSrCl2 处理后 ,三组的激活率随卵龄的增长而提高 ,分别为 2 1 1%、4 1 8%和 73 7% ,组间差异有统计学意义 (P <0 0 5 ) ;卵母细胞离体后在激活前体外培养 3h也能提高其激活率 ,但当卵龄达到一定时 (18h) ,体外培养激活率不再提高。② 5 0、10 0、15 0mmol L三种浓度的SrCl2 均能有效地激活小鼠卵母细胞 ,激活率分别为 5 8 5 %、6 8 95和 70 9% ,与对照组和 1 6mmol L组相比差异有统计学意义 (P <0 0 5 )。③卵母细胞的激活率随处理时间的延长而提高 ,30、6 0、12 0、2 4 0minSrCl2 处理时间的激活率分别为 4 4 9%、5 6 1%、6 8 9%、77 0 % ,相隔两组之间差异有统计学意义 (P <0 0 5 )。④ 1 5kV cm ,16 0 μs和 1 0kV cm ,32 0 μs脉冲刺激下的卵母细胞激活率显著高于 1 8kv cm ,90 μs脉冲刺激组 (5 8 5 %vs2 7 1%、6 9 1%vs2 7 1% ,P <0 0 5 ) ,前两组之间差异无统计学意义 (P >0 0 5 )。结论　小鼠卵母细胞孤雌激活率与卵龄和激活方案有关 ;氯化锶浓度、作用时间和     

小鼠电激活孤雌胚胎早期体内外发育能力的比较

目的 探讨小鼠电激活孤雌胚胎的早期体内、外发育能力.方法 利用不同电脉冲参数和激活液对小鼠卵母细胞进行活化,观察激活后的小鼠孤雌胚体外发育状况和移植后的发育能力.结果 非电解质激活液优于电解质液,脉冲强度、脉冲宽度和脉冲次数3个参数各自处于某一范围内时,他们之间存在某种相关性,降低其中1个参数可通过升高另外2个参数得到补偿,经筛选较适宜的电脉冲参数为:1.0 kV/cm、40μs、2p,或者1.5kV/cm、30/μs、2p,分别为74.65％和71.19％,体外囊胚发育率分别为43.40％和47.62％.电激活孤雌胚体外发育时序比正常胚胎慢,但囊胚细胞数与对照组差异不显著.它们经胚胎移植后,其中的一部分能够着床,但着床率仅为3.6％,极显著低于对照组(67％,P＜0.01).结论 电刺激能够较好地模拟正常受精过程激活小鼠卵母细胞,但激活后的多数小鼠孤雌胚胎着床能力较低,不能够顺利着床.     

细胞松弛素B和解冻方法对玻璃化冷冻小鼠卵母细胞的影响

 【目的】探讨细胞松弛素B和解冻程序对玻璃微细管法（GMP）玻璃化冷冻小鼠卵母细胞的影响。【方法】以小鼠MII期卵母细胞为模型，研究冷冻前细胞松弛素B预处理、不同解冻程序对小鼠卵母细胞冷冻效果的影响；随后对经GMP玻璃化冷冻的卵母细胞直接进行培养以检测冷冻是否诱发孤雌发育。【结果】与对照组相比．细胞松弛素B预处理的卵母细胞存活率、受精率、卵裂率及囊胚率没有显著差异（89．3％vs91．3％．44．0％vs40．4％，30．0％vs27．7％，4．0％vs6．4％；P〉0．05）；采用连续浓度梯度递减解冻法的受精率明显高于间断浓度梯度递减解冻法（57．4％vs40．4％；P〈0．05），且前者的卵裂率和囊胚发育率也相对较高f40，2％vs27，7％，14．5％vs6．4％；P〉0．05）。冷冻复苏后卵母细胞的孤雌发育率稍高于未经冷冻的卵母细胞（17．1％vs3．2％；P〉0．05）。【结论】细胞松弛素B预处理对MII期小鼠卵母细胞的GMP玻璃化冷冻保存效果没有影响：采用连续浓度梯度递减解冻法能明显提高复苏后卵母细胞的体外发育能力。     

细胞松弛素B和解冻方法对玻璃化冷冻小鼠卵母细胞的影响

【目的】探讨细胞松弛素B和解冻程序对玻璃微细管法（GMP）玻璃化冷冻小鼠卵母细胞的影响。【方法】以小鼠MII期卵母细胞为模型，研究冷冻前细胞松弛素B预处理、不同解冻程序对小鼠卵母细胞冷冻效果的影响；随后对经GMP玻璃化冷冻的卵母细胞直接进行培养以检测冷冻是否诱发孤雌发育。【结果】与对照组相比．细胞松弛素B预处理的卵母细胞存活率、受精率、卵裂率及囊胚率没有显著差异（89．3％vs91．3％．44．0％vs40．4％，30．0％vs27．7％，4．0％vs6．4％；P〉0．05）；采用连续浓度梯度递减解冻法的受精率明显高于间断浓度梯度递减解冻法（57．4％vs40．4％；P〈0．05），且前者的卵裂率和囊胚发育率也相对较高f40，2％vs27，7％，14．5％vs6．4％；P〉0．05）。冷冻复苏后卵母细胞的孤雌发育率稍高于未经冷冻的卵母细胞（17．1％vs3．2％；P〉0．05）。【结论】细胞松弛素B预处理对MII期小鼠卵母细胞的GMP玻璃化冷冻保存效果没有影响：采用连续浓度梯度递减解冻法能明显提高复苏后卵母细胞的体外发育能力。     

化学剂法对兔卵母细胞孤雌激活的效果

目的研究钙离子载体A23187和6-二甲基氨基嘌呤(6-DMAP)及3种酶对兔卵母细胞的激活率,并观察其体外发育情况。方法兔超排后获得卵母细胞,经脱颗粒处理后用A23187和6-DMAP处理不同时间,体外培养5~7d,观察细胞发育与囊胚形成情况。结果钙离子载体A23187激活处理5min后,继续在2.0mmol/L6-DMAP中分别处理3.5、4.0、4.5、5.0h,均能激活兔卵母细胞;以处理4.0h的卵裂率最高(58.82%,20/34),并有15%(3/20)的囊胚发育率。透明质酸酶、胶原酶和胰酶处理20min后,均能激活兔卵母细胞,其中透明质酸酶处理20min的激活率和卵裂率最高,分别为58.33%(28/48)和63.64%(28/44)。结论兔卵母细胞孤雌激活率与激活方法密切相关。化学激活剂的处理时间及消化酶的不同均对兔卵母细胞的激活产生显著影响。     

化学剂法对兔卵母细胞孤雌激活的效果

目的 研究钙离子载体A23187和6-二甲基氨基嘌呤(6-DMAP)及3种酶对兔卵母细胞的激活率,并观察其体外发育情况.方法 兔超排后获得卵母细胞,经脱颗粒处理后用A23187和6-DMAP处理不同时间,体外培养5～7 d,观察细胞发育与囊胚形成情况.结果 钙离子载体A23187激活处理5 min后, 继续在2.0 mmol/L 6-DMAP中分别处理3.5、4.0、4.5、5.0 h,均能激活兔卵母细胞;以处理4.0 h的卵裂率最高(58.82%,20/34),并有15%(3/20)的囊胚发育率.透明质酸酶、胶原酶和胰酶处理20 min后, 均能激活兔卵母细胞,其中透明质酸酶处理20 min的激活率和卵裂率最高,分别为58.33%(28/48)和63.64%(28/44).结论 兔卵母细胞孤雌激活率与激活方法密切相关.化学激活剂的处理时间及消化酶的不同均对兔卵母细胞的激活产生显著影响.     

小鼠卵母细胞的OPS法玻璃化冷冻保存技术

目的以小鼠卵母细胞为模型,利用开放式拉长细管(open pulled straw,OPS)法研究在不同条件下冷冻、解冻对其发育能力的影响,为家畜、濒危动物乃至人类探索一种简易、高效的卵母细胞冷冻保存提供理论与技术参考。方法在室温(25℃±0.5℃)条件下,以乙二醇(EG)、DMSO为主体抗冻保护剂配制成的玻璃化溶液(EFS、EDFS),采用OPS法对卵母细胞进行玻璃化冷冻保存。实验一:操作台温度分别为25℃±0.5℃和37℃±0.5℃时,卵母细胞在10%EG 10%DMSO或10%EG溶液中预处理30 s,移入EDFS或EFS中平衡15 s~45 s进行冷冻保存;实验二:操作台为37℃±0.5℃条件下,采用三种方法(卵母细胞在A:0.5 mol/L和0.3 mol/L的蔗糖中分别处理2 min和5 min;B:0.3 mol/L和0.15 mol/L的蔗糖中分别处理5 min和2 min;C:0.5 mol/L的蔗糖处理5 min)对EDFS30和EDFS40冷冻保存的卵母细胞进行解冻;实验三:解冻后的卵母细胞在SrCl2溶液中进行孤雌激活。结果与结论实验一中卵母细胞解冻后的形态正常率分别高达92.1%(25℃±0.5℃)和92.5%(37℃±0.5℃),均与对照组(98.9%)无显著性差异(P>0.05);实验二中卵母细胞解冻以C法效果最佳(形态正常率分别为92.5%和67.5%);实验三中解冻后在mCZB中培养0.5 h的卵母细胞激活效果优于未培养组(卵裂率和囊胚发育率分别为80.0%vs.57.7%;18.2%vs.0),均有显著或极显著差异(P<0.05或P<0.01);而培养组与新鲜对照组卵母细胞激活后卵裂率和桑椹胚发育率与对照组(80.0%vs.77.8%;52.8%vs.61.9%)均无显著性差异(P>0.05)。     

骨髓间充质干细胞条件培养液对小鼠卵母细胞的孤雌激活作用

目的 研究骨髓间充质干细胞(mesenchymal stem cell,MSC)条件培养液对小鼠MII卵母细胞的孤雌激活作用及胚胎发育能力.方法 分离、培养小鼠MSC,获取MSC条件培养液(conditioned medium of MSC,CM).通过促排技术获取小鼠MII卵母细胞,分别采用CM、7%乙醇、IVF方法激活,体视显微镜下观察原核形成及囊胚形成率.在CM激活后不同时间点,利用α/β-tubulin抗体标记纺锤体,激光共聚焦显微镜下观察有/无细胞松弛素B(CB)存在时纺锤体的运动变化.结果 CM可以激活小鼠MII卵母细胞,最佳刺激时间为40 min,激活率达到95.4%,囊胚形成率为62%,与7%乙醇组比较无显著差异,但明显低于IVF组(95.4%vs.100%;62%vs.88%,P＜0.01).CB可以抑制纺锤体的旋转,阻止第二极体的排出,促进二倍体孤雌胚形成,提高囊胚形成率(62%vs.9%,P＜0.01).结论 CM能有效激活小鼠MⅡ卵母细胞并促进孤雌发育.     

还原型谷胱甘肽对C57BL/6J小鼠体外受精的影响

目的 探讨受精液中添加还原型谷胱甘肽(GSH)对C57BL/6J小鼠体外受精的影响.方法 以体外受精液(HTF)作为对照,在HTF液中添加不同浓度(0.25 mmol/L、0.50 mmol/L、0.75 mmol/L、1.00 mmol/L、1.25 mmol/L和1.50 mmol/L)的GSH,对C57BL/6J近交系小鼠进行体外受精实验,观察体外受精和早期胚胎发育情况.同时,用5种C57BL/6J小鼠背景的基因工程小鼠加以验证.结果 与对照组相比,实验组的体外受精率显著升高,浓度为0.50 mmol/L时,体外受精率提高显著(P＜0.05),浓度在1.25 mmol/L时,体外受精率最高(P＜0.01).受精卵的体外囊胚发育率实验组与对照组无显著性差异(P＞0.05).C57BL/6J背景的基因工程小鼠的体外受精率和体外囊胚发育率与C57BL/6J近交系小鼠相似.结论 在HTF中添加一定浓度的GSH能够提高C57BL/6J小鼠及C57BL/6J背景的基因工程小鼠的体外受精效果.     

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.     

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…     

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.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

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.