组织外植块对培养的鸡胚背根节神经突起生长的影响

为了解神经元的发育和周围组织的关系,本文分别用:(1) 10天鸡胚背根节与10天鸡胚心脏、皮肤、角膜、骨骼肌、肠、大脑、脊髓外植块联合培养;(2) 不同时期的鸡角膜与不同胚龄的鸡胚背根节联合培养;(3) 10天鸡胚背根节与雏鸡角膜内、外层联合培养。用于联合培养的背根节和组织外植块均按Maximow双盖片法种植在涂有鼠尾胶原的盖玻片上,两者相距1.5mm。外植块大小约0.5×1×1mm~3。培养48小时后取出固定,用改良的Bodian蛋白银法浸镀染色,在低倍镜下对神经突起进行半定量计数。 实验结果表明鸡胚心脏,皮肤和角膜外植块对背根节神经突起生长具较强的促进作用且对生长方向有明显诱导作用;骨骼肌、肠和大脑对神经突起生长也有不同程度促进作用,但脊髓却无明显作用。14、16、18天鸡胚角膜对8、10、12、14天鸡胚背根节神经突起生长有明显促进作用;12、14天鸡胚背根节在各时期角膜作用下神经突起生长都较丰富。含上皮层的雏鸡角膜外植块促进背根节神经突起生长的作用比含内皮层的角膜外植块要强。     

人神经营养素-3成熟区基因在大肠杆菌内的表达

【目的】应用基因工程方法制备人神经营养素 3(hNT 3)蛋白 ,为探讨其对阿尔茨海默病等中枢神经退行性疾病的治疗作用提供材料。【方法】将通过序列分析确定的hNT 3成熟区基因重组至原核表达质粒pBV2 2 0中 ,构建了重组表达载体 pBV/mhNT 3 ,在大肠杆菌中表达后 ,SDS PAGE法测定表达蛋白的相对分子质量 ,鸡胚背根节培养法检测其生物学活性。【结果】pBV/mhNT 3可在大肠杆菌中表达出一相对分子质量为 15× 10 3 的新蛋白 ,与hNT 3蛋白相对分子量相符。表达蛋白主要以包涵体形式存在 ,经纯化复性后 ,可明显促进鸡胚背根节的生长。【结论】人神经营养素 3成熟区基因可在大肠杆菌内高效表达hNT 3。     

腺苷酸环化酶激活剂Forskolin促背根节神经突起生长作用的研究

为探讨Ｆｏｒｓｋｏｌｉｎ对感觉神经元神经突起生长的影响,采用悬浮培养法,对鸡胚背根节进行体外培养。结果：培养２４ｈ后,对照组、Ｆｏｒｓｋｏｌｉｎ组、神经生长因子（ＮＧＦ）组及Ｆｏｒｓｋｏｌｉｎ加ＮＧＦ组生长的神经突起的平均长度分别为９１．２±１７．３μｍ、２７２．３±３１．４μｍ、２９５．３±２８．７μｍ、３０１．８±４０．１μｍ。与对照组相比,三个实验组生长的神经突起的平均长度均较长（Ｐ值均＜０．０１）;Ｆｏｒｓｋｏｌｉｎ加ＮＧＦ组与其它二个实验组比较,神经突起的平均长度无显著差异（Ｐ值均＞０．０５）。结果提示：１、Ｆｏｒｓｋｏｌｉｎ与ＮＧＦ均有促鸡胚背根节神经突起生长的作用;２、Ｆｏｒｓｋｏｌｉｎ与ＮＧＦ无协同促鸡胚背根节神经突起生长作用。     

坐骨神经源性因子对培养中成年青蛙背根节神经元的作用

【目的】探讨离断的坐骨神经 (sciaticnerve,SN)分泌的可溶性因子对培养中的成年青蛙背根神经节 (dorsalrootganglion ,DRG)神经突起生长及形态学的影响。【方法】分离成年青蛙DRG神经元培养于含有离断的SN条件性培养基(SNCM)中 ,检测SNCM促进神经突起生长的量效和时效作用及对生长的神经突起形态学的影响。【结果】在 0 1～ 5mg/L蛋白质量浓度范围内 ,SNCM促进神经突起生长的作用有蛋白质量浓度依赖关系。在培养的第 1天 ,实验组平均神经突起长度是对照组的 3 9倍 ,培养 3d后实验组神经元突起平均长度已达对照组神经元的 9 2倍 ,实验组和对照组两者有显著的差异 (P <0 0 0 1)。且实验组神经元生长出的突起缺乏对照组神经元那种典型的片层状伪足。活性因子的分子质量介于 30～10 0ku之间。【结论】离断的外周神经可释放可溶性的因子促进离体培养的成年DRG神经元突起的生长并影响突起的形态。     

体外培养鸡胚背根节神经细胞的形态学观察

为探讨鸡胚背根节(DRG)神经元的体外发育过程,分别取培养0、 1、 2、 3、 4、 5、 7、 14、 21天的神经元,计数相同面积内(2.5×105μm2)神经元的存活数,观察培养神经元的形态变化并与体内者比较.结果 :各时相神经元的存活数分别是94±4、 83±6、 67±4、 65±7、 68±5、 71±9、 73±6、 22±4、 0 个.神经元的形态在培养48小时内无显著变化,多为球形或椭圆形,但至3天后,不少神经元呈锥形或梭形,而体内者仍为圆球形或卵圆形.由此提示,离体后,仅部份神经元在一定时段内能够存活生长,且神经元的体外发育过程可能与其在体内的情况有所不同.     

低相对分子质量透明质酸的促血管生成作用

目的 研究低相对分子质量透明质酸(LMWHA)的促血管生成作用.方法 采用鸡胚绒毛尿囊膜(CAM)血管生成模型,观察不同相对分子质量(Mr)的LMWHA及不同给药剂量对血管生成的影响.结果 Mr低于10&#215;103的LMWHA对CAM血管生成的影响与溶剂对照组相比,差异有非常显著性意义(P＜0.01);随着给药剂量的增大,新生血管总数明显增多(Mr6 000,r=0.992 9;Mr6 240,r=0.994 4).结论 Mr低于10&#215;103的LMWHA具有促进CAM血管生成作用,且呈剂量依赖性.     

人睫状神经营养因子基因的克隆、表达、纯化和生物活性鉴定

目的表达和纯化人睫状神经营养因子(CNTF)并观察其生物学活性。方法用PCR方法构建人CNTF的结构基因,并克隆于pLy4原核表达载体中。将重组表达质粒pLy4-CNTF转化大肠埃希菌JF1125,30℃培养至A600(nm)达到0.6时,迅速升温至42℃诱导目的蛋白的表达。细菌经超声破胞,包涵体经变性、复性、透析和阴离子交换层析,获得纯化的CNTF重组蛋白。然后,用鸡胚背根神经节神经元细胞存活实验观察其生物活性。结果42℃诱导后可以获得Mr≈22.9×103的目的蛋白,Western印迹结果进一步表明,其具有人CNTF的抗原性。表达蛋白主要以不溶形式存在,占菌体蛋白的35%以上,表达产物经变性、复性,纯化后可获得纯度90%以上的目的蛋白。该重组蛋白能够促进鸡胚背根神经节神经元细胞的存活。结论在大肠埃希菌JF1125中成功表达了人CNTF分子,经变性、复性,纯化后得到具有生物活性的蛋白。     

一种简单高效的新生鼠海马神经元原代培养方法

【目的】建立一种简单、高效、高纯度的新生小鼠海马神经元的原代培养方法。【方法】取出生24 h内的C57BL/6J新生鼠,分离海马组织,采用胰蛋白酶消化加机械吹打的方法获得单个细胞,用含体积分数1%B27及2%Glutamax-100X、终浓度25μmol/L Glutamate的Neurobasal-A培养基接种培养,3 d后用去Glutamate成分的上述培养基换液,并加阿糖胞苷作用48 h以抑制胶质细胞增长。于倒置相差显微镜下观察细胞的生长状态,采用微管蛋白相关标志物2(MAP2)及Hoechst33258免疫荧光染色鉴定神经元纯度。【结果】此培养方法获得的海马神经元生长状态良好,细胞形态从接种时的圆形透亮、无突起,到培养第7天后发展为神经元胞体聚集、树突发达并形成纵横交错的神经网络;经鉴定,神经元的纯度高达97.2%以上,且能稳定存活2~3周。【结论】该方法操作简单、高效,所得神经元纯度高,结果稳定。     

明胶作载体对NGF生物活性的影响

作者将明胶＋神经生长因子（Nervegrowthfactor，NGF），单纯NGF，明胶和单纯0．9％生理盐水4种物质分别：①加到无血清培养基中，观察对鸡胚背根神经节突起生长的影响．②注入大鼠坐骨神经12mm缺损的再生硅胶管小室内，术后5周、8周、12周进行光镜、电镜、图像分析等检测．结果：明胶＋NGF和单纯NGF一样可促进鸡胚背根神经节的存活并促进其细胞突起的生长和大鼠坐骨神经再生，证明明胶对NGF的生物活性无影响．     

己烯雌酚对胎鼠海马神经干细胞增殖与分化的影响

目的:探讨己烯雌酚对胎鼠海马神经干细胞(neural stem cells,NSCs)的促增殖、分化作用及其机制。方法:取孕16天SD胎鼠海马NSCs培养,分别设空白对照组,己烯雌酚1×10-7 mol/L组、1×10-8 mol/L组、1×10-9 mol/L组,以及脑源性神经营养因子(BDNF)5×10-2 mg/L组;免疫荧光化学方法检测Nestin、神经元特异烯醇化酶及神经胶质酸性蛋白的表达,RT-PCR检测BDNF mRNA的含量。结果:不同剂量己烯雌酚处理各组神经球面积、积分光密度值、平均突起长度与平均突起数均增加,其中10-8 mol/L组海马NSC增殖与分化最明显,BDNF mRNA表达量亦最高(P<0.01)。结论:己烯雌酚对海马NSCs的增殖分化具有促进作用,且可能与上调BDNF的表达有关。     

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…