大鼠脑微血管内皮细胞的培养与鉴定

目的探讨大鼠脑微血管内皮细胞体外培养方法,为血管内皮细胞的研究打下基础。方法用胶原酶消化大鼠脑皮质制成细胞悬液再进行培养,用光镜及免疫组织化学检测进行鉴定。结果分离的大鼠脑微血管内皮细胞体外培养5～7d左右可长成单层,光镜下为多角形或扁平梭形,呈"铺路石样"排列,第Ⅷ因子相关抗原免疫组织化学测定阳性。结论该分离细胞培养法可培养出大鼠的脑微血管内皮细胞。     

大鼠脑微血管内皮细胞的原代培养

目的分离、培养原代脑微血管内皮细胞,建立体外血脑屏障模型。同时探索高纯度、活性好的脑微血管内皮细胞的分离培养方法。方法取3周大鼠大脑,分离皮质,经过匀浆、葡聚糖离心以及消化后,取纯度较高的脑微血管段种植于胶原蛋白包被过的塑料培养瓶中进行培养。显微镜观察及检测Ⅷ因子相关抗原。结果镜下细胞呈长梭形,7d左右细胞可融合,Ⅷ因子相关抗原免疫组化检测为阳性,且阳性细胞占绝大部分。结论本实验成功分离大鼠脑微血管内皮细胞,并进行原代培养,为脑微血管内皮细胞的进一步研究提供了模型,也为构建更高级的大鼠体外血脑屏障奠定基础。     

兔脑微血管内皮细胞的培养及鉴定

目的：建立简便、有效的兔脑微血管内皮细胞体外原代培养方法。方法：兔脑皮质通过匀浆、过筛分离处理后接种，原代培养兔脑微血管内皮细胞，并通过Ⅷ因子相关抗原免疫组化染色和透射电镜观察鉴定。结果：培养的细胞呈典型的“铺路石”状生长，Ⅷ因子相关抗原免疫组化染色为阳性。结论：该方法可方便获取较纯净的脑微血管内皮细胞。     

大鼠脑微血管内皮细胞的分离、培养及不同纯化方法的比较

目的 对脑微血管内皮细胞分离、培养方法进行探索性研究,并对目前常用的纯化方法进行比较.方法 首先采用两次酶消化、两次梯度离心法对大鼠脑血管内皮原代细胞进行分离,然后分别采用免疫磁珠和Thy1.1抗体杀伤法分别进行纯化.结果 采用两次酶消化、两次梯度离心法可以成功分离到脑微血管内皮细胞,磁珠纯化法可得到纯度高但是数量较少的细胞,Thy1.1抗体杀伤法可得到纯度及得率均较高的内皮细胞.结论 我们所摸索的方法能成功进行纯度较高的大鼠脑微血管内皮细胞原代培养.     

脑溢安对缺氧大鼠脑微血管内皮细胞VEGF蛋白表达的影响

目的:观察脑溢安血清对体外缺氧培养的大鼠脑微血管内皮细胞血管内皮生长因子(VEGF)蛋白表达的影响。方法:大鼠用脑溢安浸膏连续灌胃3d后,心脏内采血分离出血清;用新生7d SD大鼠脑内分离培养的微血管内皮细胞（RCMEC）移入厌氧培养箱造成缺氧模型;MTT法测定细胞活性;免疫细胞化学及Werstern blotting研究脑微血管内皮细胞VEGF蛋白表达。结果:缺氧18h脑溢安组OD值较对照组增高（P<0.05）。缺氧损伤的脑微血管内皮细胞内VEGF蛋白表达增强,脑溢安血清组VEGF蛋白的表达水平高于对照组(P<0.01)。结论:缺氧可诱导脑微血管内皮细胞VEGF蛋白的表达;脑溢安可以上调VEGF蛋白的表达,这可能是脑溢安对缺氧的脑微血管内皮细胞的保护作用机制之一。     

大鼠原代脑微血管内皮细胞体外分离与培养的实验研究

目的 探讨获取大鼠脑微血管内皮细胞(BMEC)的简单、有效方法,为构建体外血肿瘤屏障(BTB)模型提供材料.方法 采集出生3～5 d的Wistar胎鼠大脑皮质,应用酶消化法及葡聚糖离心法获得脑微血管段后,接种于培养皿中进行原代培养,采用倒置显微镜对所培养的细胞进行形态学观察;以Ⅷ因子相关抗原免疫组化染色法鉴定细胞;将BMEC与C6脑胶质瘤细胞共培养,构建体外BTB模型,并采用免疫组化法和免疫荧光法检测BMEC间紧密连接相关蛋白occludin的表达.结果 体外培养2h时脑微血管段贴壁,12～48 h见圆形生发中心形成,2～3d单层内皮细胞自生发中心长出,4～5 d见较大单层内皮细胞团,5～7 d可见融合成片的内皮细胞单层,外观呈“铺路石”样;第Ⅷ因子免疫组化结果显示“铺路石”样细胞胞质呈棕黄色染色;紧密连接相关蛋白occludin的免疫组化和荧光结果证明共培养的BMEC间表达BTB的特性.结论 本方法能成功地进行大鼠原代BMEC培养,构建大鼠体外BTB模型,进而应用于BTB的生理、生化及药理学研究.     

大鼠脑微血管内皮细胞的体外培养方法

目的建立一种简便可靠的脑微血管内皮细胞的培养方法.方法新生SD乳鼠脑皮质经匀浆、过滤、消化和差速贴壁等方法对大鼠脑微血管内皮细胞进行原代培养及传代培养,对培养的细胞进行形态学观察,并用Ⅷ因子相关抗原进行免疫细胞化学鉴定.结果培养的细胞呈单层贴壁生长,约7～9d后呈典型的"铺路石"征象,免疫细胞化学鉴定证实为内皮细胞.结论此种脑微血管内皮细胞培养方法的建立为体外研究脑血管疾病提供了可靠的手段.     

小鼠肺微血管内皮细胞培养及鉴定

目的:建立小鼠肺微血管内皮细胞体外分离培养方法及对微血管内皮细胞的鉴定.方法:取小鼠肺组织边缘剪成小块,用含有20%新生牛血清、肝素90μg/mL、L-谷胺酰胺4 mmoL/L、青霉素200 U/mL和链霉素200μg/mL的RPMI-1640培养基进行培养.纯化细胞进行Ⅷ因子相关抗原间接免疫组化染色和透射电镜观察Weilel-palade小体.结果:获得的内皮细胞具有规律的鹅卵石样形态,Ⅷ因子相关抗原间接免疫组化染色阳性而且在透射电镜观察到Weilel-palade小体.结论:成功建立了小鼠肺微血管内皮细胞体外培养方法,并为体外研究肺微血管内皮细胞提供实验模型.     

前列腺毛细血管内皮细胞屏障功能研究

目的 分离培养大鼠前列腺毛细血管内皮细胞,体外研究大鼠前列腺毛细血管内皮细胞的屏障功能.方法 采用细胞生长特性选择法分离培养大鼠前列腺毛细血管内皮细胞,用第Ⅷ因子相关抗原抗体对分离培养的前列腺毛细血管内皮细胞进行免疫学鉴定.免疫组化观察毛细血管内皮细胞之间紧密连接蛋白claudin-1的表达,采用跨膜电阻测量仪检测毛细血管内皮细胞的跨膜电阻,用酚红渗漏实验检测其通透性,以大鼠肺毛细血管内皮细胞、脑毛细血管内皮细胞作为阴性与阳性对照.结果 成功分离大鼠前列腺毛细血管内皮细胞,体外培养可以形成紧密的单层细胞结构,呈铺路石样生长.第Ⅷ因子相关抗原抗体的免疫荧光结果显示:分离培养的大鼠前列腺毛细血管内皮细胞胞浆表达绿色荧光,提示分离的细胞为毛细血管内皮细胞;免疫组化结果显示紧密连接蛋白claudin-1在相邻的毛细血管内皮细胞之间表达;体外培养的大鼠前列腺毛细血管内皮细胞,阴性对照组肺毛细血管内皮细胞,阳性对照组脑毛细血管内皮细胞的跨膜电阻峰值分别可以达到(142.2 ±3.1)、(43.3±3.5)、(248.2±6.2)Ω/cm2,并稳定保持2～3d,三者比较差异有统计学意义(P＜0.05).酚红渗漏实验结果显示前列腺毛细血管内皮细胞、阳性对照组脑毛细血管内皮细胞随着细胞单层跨膜电阻的增加,基底侧的酚红浓度减低,阴性对照组肺毛细血管内皮细胞酚红渗透量随跨膜电阻的改变变化趋势不明显.结论 大鼠前列腺毛细血管内皮细胞与脑毛细血管内皮细胞等屏障内皮相比拥有相似的结构功能,体外培养的大鼠前列腺毛细血管内皮细胞具有屏障特性,可以作为血前列腺屏障研究的体外模型.     

人卵巢癌微血管内皮细胞的分离培养与鉴定

目的建立稳定可靠的卵巢癌微血管内皮细胞分离培养方法.方法取上皮性卵巢癌组织,酶消化,经Percoll分离介质梯度离心得到单个核细胞进行培养,Ⅷ因子相关抗原免疫组化和透射电镜观察鉴定.结果接种后24 h,细胞开始贴壁,呈多边形或短梭形,8～10 d细胞融合成单层片状,呈现出"铺路石样"外观.Ⅷ因子相关抗原免疫组化染色阳性,透射电镜下可见W-P小体.结论本研究方法能够获取体外培养的卵巢癌源性微血管内皮细胞.     

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…