精子膜处理方法对精子介导的外源质粒转基因效率的影响

目的 探讨超声、NaOH和溶血卵磷脂三种小鼠精子膜处理方法对转染质粒与精子共孵育后卵胞质内单精子注射(ICSI)授精,观察质粒转染的效率.方法 采用上述三种方法对小鼠精子进行膜处理,通过Transgreen/PI荧光染色,观察小鼠精子质膜的完整性;通过流式细胞仪检测小鼠精子DNA的完整性;将处理后的小鼠精子与带GFP的转染质粒共孵育,再采用ICSI与促排卵后成熟小鼠卵子授精,统计各组受精率、卵裂率及荧光囊胚率,观察三种方法对小鼠精子作为质粒载体ICSI后转基因效率的影响.结果 溶血卵磷脂方法处理小鼠精子膜的效果显著优于超声和NaOH组(99.2％ vs 94.7％％,90.9)(P＜0.05).三种方法对小鼠精子DNA结构的损伤无显著性差异(P＞0.05),行ICSI后三组小鼠胚胎发育率差异不显著(P＞0.05),各组之间荧光胚胎的形成率无显著性差异(P＞0.05).结论 三种小鼠精子膜处理方法的效率均能达到90％以上,且对小鼠精子DNA的损伤、行ICSI后小鼠胚胎的发育率和转基因效率无显著差异,因此,三种方法均可用于ICSI法小鼠转基因研究.     

睾丸内注射和体内电穿孔法建立转基因小鼠的实验研究

目的 探讨在体睾丸生精小管注射合并体内电穿孔建立转基因小鼠的可行性.方法 将两种不同处理的转染液分别注射到4组SPF级雄性昆明小鼠睾丸生精小管内,然后对同一种转染液注射的两组动物中的一组进行体内电穿孔,另一组则不进行体内电穿孔;2周后各组SPF级雄性昆明小鼠分别与超排处理的SPF级雌性昆明小鼠合笼交配,最后对各组新生仔鼠进行PCR-Southern检测.结果 PCR检测,四组之间存在显著差异(P＜0.01),并且以A组最高(45%);Southern blotting检测,4组之间差异不显著(P＞0.05),但A组的整合效率(25%)仍高于其他3组.结论 运用在体睾丸生精小管内注射合并体内电穿孔技术可以显著地提高外源基因的转染效率,该方法是否可广泛用于生产转基因动物还有待于进一步深入研究.     

胞内精子注射法制备转基因爪蟾的研究

目的研究爪蟾胞内精子注射法(intracytoplasmic sperm injection,ICSI)转基因技术的可行性。方法取成熟雄蛙的睾丸分离纯化精子,用毛地黄皂甙(digitonin)崩解精细胞膜并制备精子浓缩液,然后与线性化报告基因载体pCMV-EGFP-N1共处理,最后将处理后的精子注射入从雌性爪蟾体内取出的未受精卵中,培养并观察。结果制备的精子质量较高,稀释的精子注射入未受精卵后,卵的受精率为10%,受精卵活过神经胚的比率为20%,但都略微有点畸形。其中,绿色荧光报告基因EGFP的整合率为81%,但镜下未见绿色荧光蛋白的表达。结论ICSI法是制备转基因爪蟾的简便可行的新途径。     

精子注射IVF技术与后代发育迟缓无关

一份新的研究结果指出,经在胞质内注射精子(ICSI)而怀孕分娩的孩子不会增加息神经发育迟缓的风险。 伦敦皇家自由大学医学院的Dr.Alastair G.Sutcliffe和他的同事们认为,有几种可能的原因来解释ICSI对发育的影响。理论上使用不太成熟的精子可能会发生基因异常或结构缺损。还有,技术本     

既往卵母细胞单精子显微注射(ICSI)不良结局和有限的第2次有丝分裂中期卵细胞的患者应用激光辅助的ICSI

在本研究中,激光辅助的卵母细胞单精子显微注射(ICSI)改善了既往ICSI不良结局和有限的第2次有丝分裂中期卵细胞的患者受精率和受精卵质量。这项技术在操作过程中对卵子创伤小,可推广使用。既往卵母细胞单精子显微注射(ICSI)不良结局和有限的第2次有丝分裂中期卵细胞的患者应用激光辅助的ICSI@Demirol A.$Women's Health, Infertility and IVF Center, Ankara, Turkey Dr. @Benkhalifa M. @Sari T. @Gurgan T. @张旸…     

转基因的方法

显微注射法和胚胎干细胞转染法是制作转基因动物的两种主要方法。这两种方法的目的是制作一种人工改变基因型的动物。两种方法之间的差异和其技术限度决定了各自的实验用途。因此,阐述这两种方法并正确评估两者的应用是很有价值的。显微注射原核制作转基因小鼠如同Gorden等(1980)报道过的,标准方法是显微注射小鼠胚胎。取交配后第2天的小鼠胚胎,将胚胎清洁后放置于操作显微镜。在此阶段两个配子融合,但细胞核(称为原核)仍     

提高制备转基因小鼠效率的研究

目的提高制备转基因动物的效率.方法着重对注射DNA的制备,高质量受精卵的获得,显微注射及胚胎移植各步骤中的主要影响因素进行了优化.结果移植后出生的小鼠经PCR及Sonthern-Blot检测,从最初的146只仔鼠中检测到2只阳性鼠,提高到由34只仔鼠中检测到10只阳性鼠,阳性鼠概率由1.4%基本稳定增加到30%.结论通过技术改良后,转基因效率得到明显提高,为从事转基因动物方面的研究建立了稳定的技术平台.     

常规体外受精-胚胎移植与卵细胞浆内单精子注射临床效果比较

目的 :比较常规体外受精 -胚胎移植 ( in vitro fertilization and embroy transfer,IVF- ET)与卵胞浆内单精子注射 ( intracytoplasmic sperm injection,ICSI)两种助孕技术的临床效果。方法 :61对夫妻进行 61个周期 ( IVF31周期 ,ICSI30周期 ) ,比较两组每周期平均获卵数、正常受精卵数、卵裂数、移植胚胎数、生化妊娠率和临床妊娠率。结果 :ICSI组平均每周期平均获卵数、正常受精卵数、卵裂数、移植胚胎数与 IVF组相似 ( ICSI组为 9.31、6.40、5 .63、2 .71 ;IVF组为 1 0 .0 2、7.5 3、5 .91、2 .69。 P>0 .0 5 ) ;ICSI组平均每周妊娠率与IVF组相似 ( ICSI组 36.7% ,IVF组 38.7%。 P>0 .0 5 )。结论 :ICSI技术治疗严重男性因素引起的不育症与常规 IVF治疗女性因素引起的不孕症临床结果相似。     

大鼠精子冷冻保存和卵胞浆内精子注射研究进展

许多哺乳类动物精子冷冻保存技术均已成功建立。然而,成功建立一套啮齿类动物精子冷冻保存程序还具有一定的挑战性。尤其是大鼠,因为其较长的精子尾巴,使得它的精子冷冻保存相当困难,并且大鼠的卵胞浆内精子注射(ICSI)技术也没有小鼠那样成熟。国内尚未见关于大鼠精子冷冻保存和ICSI的报道,国际上关于这方面的报道也较少。本文就这两方面的研究进展做一简要综述。     

体细胞核移植技术进展

转基因动物的研究和生产是动物胚胎工程中最诱人和最有发展前景的课题之一。早期采用原核内注射外源基因的方法，但整合效率低，出生的转基因动物外源性基因的表达也不稳定。核移植是近年发展起来的一种细胞克隆技术。本文介绍了核移植的定义、细胞去核方法，着重对核移植的三种方法电融合法、PEG介导法、微注射法的原理、应用、优缺点比较等研究进展进行综述。核移植技术在生产转基因动物、适于移植杩细胞克隆、器官等方面有广泛的应用前景。     

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.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

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.