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。     

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…     

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.     

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.     

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.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.     

Survey and practice of reporting quality of randomized controlled clinical trials on traditional Chinese medicine

Evidence obtained from randomized controlled trials （RCTs） has been generally accepted as the gold standard in the evaluation of clinical effectiveness. Readers need to understand the trial design, implementation, results, analysis and interpretation, so as to fully Jnderstand the results of RCTs. Thus, the investigators of RCTs have to report these items in a complete, accurate and clear manner. Since 1998, we have conducted several evaluations on the reporting quality of RCTs published in Chinese journals on traditional Chinese medicine （TCM） and results have shown that there is an urgent need for higher quality RCTs on TCM.     

TCM Treatment of Ankylosing Spondylitis by Tonifying the Kidney and Strengthening the Governor Vessel

Ankylosing spondylitis is a chronic and progressive disorder with inflammation mainly involving the central axis joints. It mainly affects the cervical spine and the lumbosacral area, with the pathogenesis closely related to the kidney and the Governor Vessel (GV). TCM holds that the syndrome is deficiency in origin and excess in superficiality, which is due to insufficiency of the kidney, deficiency of GV, and blocking of the channels with the invasion of exogenous evil, leading to poor circulation of qi and blood and malnutrition of the bones, muscles and joints. The TCM method of tonifying the kidney and strengthening GV to regulate circulation of qi and blood and check the arthralgia pain should be adopted, with the Kidney-Tonifying and GV Strengthening Decoction (益肾强督汤) prescribed.     

IMPROVING P-gp EXPRESSION IN HUMAN MONONUCLEAR CELLS IN VITRO TRANSFECTED BY MULTIDRUG RESISTANCE-1 mRNA

CHEMOTHERAPY playsa greatrolein the treat- ment of malignanttumors,especiallyingynecolo- gicalones.But inanticancerchemotherapy,leuko-cytopeniaisfrequentlytheprimarydose-limitingsideeffect factor.Moreover,cancersarefrequentlychemoresistantbe-causeof overexpressionof P-glycoprotein(P-gp), which isencodedby multidrugresistancegene (MDR1 ) and detectableinup to50% ofhuman cancersand renderscellsresistancetoanticancerdrugs.The safetyand potentialtherapeuticbenefitof mdr1 gene transferredto h…