表皮干细胞在人体不同部位皮肤的分布差异

目的:研究表皮干细胞在人体不同部位皮肤的分布差异.方法:从5例成年男性志愿者身体取头部、胸部、背部、臀部、大腿内侧、大腿外侧、上臂内侧、上臂外侧、手掌、足底、包皮及阴囊皮肤标本,采用免疫组织化学EnVision法检测皮肤组织表皮干细胞标志物角蛋白19(K19)和整合素β1的表达.结果:皮肤基底层K19和整合素β1阳性细胞以包皮和阴囊最多,其次是臀部,背部及四肢近端外侧相应地多于胸部及四肢近端内侧,头皮、手掌及足底皮肤基底层阳性细胞很少.头皮毛囊隆突部及皮下腺管可见较多K19和整合素β1阳性细胞.结论:表皮干细胞在人体不同部位皮肤的分布存在差异,头皮的毛囊隆突部及包皮和阴囊的皮肤基底层存在较多表皮干细胞,背部及四肢近端外侧皮肤基底层表皮干细胞相应地多于胸部及四肢近端内侧.     

表皮细胞生长因子治疗创面出现的干细胞岛现象

目的：从细胞分化角度研究皮肤干细胞分布与增殖分化特征,以及这些特征在加速受创皮肤再生中的作用。方法：取8例经重组人表皮细胞生长因子（rhEGF)治疗而愈的溃疡他是面皮肤,7例经磺胺嘧啶银治疗而愈的创面皮肤以及3例流产胎儿、3例儿童和2例正常成人皮肤,用过氧化酶标记的链霉卵白素（SP）免疫组化法研究β1整合素、角蛋白19（K19）、角蛋白14（K14）以及角蛋白10（K10）的表达特征。结果：免疫组织化学染色显示,经rhEGF治疗的创面表皮干细胞和短暂扩充细胞不仅数量多,而且体积增大,在再生表皮基底层与角质层之间出现若干干细胞岛。这些干细胞岛的特征是β1整合素和K19染色阳性,在组织学上与基底层干细胞没有直接联系,为一类独立的岛状结构。经K14与K10等系列染色鉴别研究表明,在正常成人、青少年、胎儿以及未用rhEGF治疗的对照创面皮肤均未发现类似的干细胞岛现象。结论：在经rhEGF治疗的创面表皮中出现的干细胞岛是一种特殊和独立的细胞学结构,可能由已经分化成熟的细胞逆分化而来。其机制涉及生长因子对细胞分化调控等诸方面。     

成人体皮表皮干细胞的定位及分离培养

目的:研究表皮干细胞在成人体皮中的分布及其分离和培养.方法:用免疫组织化学方法检测成人体皮中β1整合素及角蛋白19的表达,对成人体皮中的表皮干细胞进行定位和定量;用Ⅳ型胶原分离培养成人体皮表皮干细胞,通过检测β1整合素及角蛋白19的表达水平及克隆形成率(以角质细胞为对照)对其进行鉴定.结果:β1整合素及角蛋白19的免疫组化阳性信号主要位于表皮的基底部及毛囊周围,阳性细胞在表皮中所占的数量较少,且在真皮中也有少量表达.体外培养,可见细胞呈克隆样生长、β1整合素及角蛋白19免疫组织化学染色呈阳性,且其克隆形成率为17.17%,明显高于对照组的6.83%(P＜0.05).结论:应用Ⅳ型胶原快速贴附法及成人成纤维细胞条件培养液,对成人体皮中表皮干细胞成功进行了分离培养,为表皮干细胞体外的大量扩增奠定了基础.     

毛囊干细胞的体外研究

毛囊是皮肤的重要附属器官,其生长具有周期性.这与其中存在的毛囊干细胞的功能密不可分.毛囊干细胞具有强大的增殖潜能及多向分化能力,可以分化成毛囊及表皮的多种细胞.它不仅对维持毛发的周期性生长有重要作用,同时也是表皮及皮肤附属器官再生的重要细胞来源.目前对成人毛囊干细胞的定位还没有明确的结论,多数的在体研究认为它可能存在于毛囊膨出部(立毛肌插入点附近)和外根鞘的最外层,其特点是体积小,分化程度低,具有慢周期性,特异性表达β1整合素和角蛋白19型(Keratin 19,K19).     

ES细胞源性表皮干细胞与胶原海绵构建组织工程皮肤

【目的】以胚胎干细胞（ES细胞）源性表皮干细胞为种子细胞与胶原海绵构建组织工程皮肤，探讨其对皮肤缺损修复的作用。【方法】胎鼠皮肤成纤维细胞与胶原海绵构建类真皮，植入小鼠全层皮肤缺损创面。以生物膜为载体，把羊膜诱导后带有核标记的ES细胞源性表皮干细胞覆盖在类真皮上，术后1～8周连续取材，HE染色，B1整合素、CK15、CK19、CK10和CEA免疫荧光双标和免疫组化观察。【结果】植入后3周，创面完全长合。较厚新生皮完全覆盖创面，基底层细胞增生，形成许多大小不一的细胞柱伸向真皮层，新生表皮中可见核标记的细胞呈B1整合素、CK15、CK19阳性，真皮中的管腔样结构呈核荧光和β1整合素、CEA免疫组化双标阳性，4～8周新生表皮基底层细胞呈CK19、CK10阳性，新生表皮下可见毛囊样、皮脂腺样结构。【结论】ES细胞源性表皮干细胞为种子细胞与胶原海绵构建的组织工程皮肤可以修复缺损皮肤，在其下真皮层有分化为毛囊样、皮脂腺样和汗腺样的结构，但是是否来源于ES细胞源性表皮干细胞仍需进一步证实。     

ES细胞源性表皮干细胞与胶原海绵构建组织工程皮肤

【目的】以胚胎干细胞(ES细胞)源性表皮干细胞为种子细胞与胶原海绵构建组织工程皮肤,探讨其对皮肤缺损修复的作用。【方法】胎鼠皮肤成纤维细胞与胶原海绵构建类真皮,植入小鼠全层皮肤缺损创面,以生物膜为载体,把羊膜诱导后带有核标记的ES细胞源性表皮干细胞覆盖在类真皮上,术后1～8周连续取材,HE染色,β1整合素、CK15、CK19、CK10和CEA免疫荧光双标和免疫组化观察。【结果】植入后3周,创面完全长合,较厚新生皮完全覆盖创面,基底层细胞增生,形成许多大小不一的细胞柱伸向真皮层,新生表皮中可见核标记的细胞呈β1整合素、CK15、CK19阳性,真皮中的管腔样结构呈核荧光和β1整合素、CEA免疫组化双标阳性,4～8周新生表皮基底层细胞呈CK19、CK10阳性,新生表皮下可见毛囊样、皮脂腺样结构。【结论】ES细胞源性表皮干细胞为种子细胞与胶原海绵构建的组织工程皮肤可以修复缺损皮肤,在其下真皮层有分化为毛囊样、皮脂腺样和汗腺样的结构,但是是否来源于ES细胞源性表皮干细胞仍需进一步证实。     

大鼠表皮基底层干细胞体外分离与培养

目的建立简单、可靠的大鼠表皮基底层干细胞体外分离培养方法。方法取新生大鼠背部皮肤,采用胰酶两步消化法获得单细胞悬液,Ⅳ型胶原差速贴壁法富集干细胞,将慢黏附细胞作为对照组细胞,均以角质形成细胞无血清培养基培养。以β1-整合素和角蛋白19(K19)双重荧光染色鉴定细胞表型,克隆形成实验检测细胞体外增殖能力。结果分离培养的细胞为多角形、呈铺路石样排列,倍增时间约为24h,细胞形态和生长规律符合基底层干细胞的特征。免疫荧光鉴定显示细胞共表达β1-整合素和K19,基底层干细胞克隆形成能力显著强于对照细胞。结论两步消化联合Ⅳ型胶原差速贴壁法获取基底层干细胞简易可行,培养的细胞活力好、表型可靠。     

正常皮肤和新生疤痕组织表达角蛋白19的比较性研究

目的:研究正常皮肤和新生疤痕组织中表皮干细胞分布、增殖分化特征,并探讨这些特征与皮肤创伤修复的关系。方法:选择有Ⅲ度烧伤创面的成人患者8例,在获得了患者的知情同意后用手术尖刀分别切取正常皮肤、肉芽中的皮丁及新生疤痕组织(创面愈合后2个月内的疤痕)各一块,约2cm×1cm×0.5cm大小,应用过氧化酶标记的链霉卵白素(SP)免疫组化法,以小鼠抗人角蛋白19型的单克隆抗体检测表皮干细胞的分布、增殖分化特征。结果:肉芽中的皮丁组织内有散在分布的角蛋白19表达阳性细胞,新生疤痕组织中的表皮基底层有多层角蛋白19表达阳性细胞,在再生表皮的中部,即位于基底细胞层与角质细胞层之间有大量的角蛋白19表达阳性细胞,且分布广泛,而正常皮肤只有表皮基底层和附件有角蛋白19表达阳性细胞。结论:新生疤痕组织中表皮再生仍很活跃,在皮肤创伤愈合过程中的重塑期,如果能够结合抑制疤痕增生的最有效治疗,这种新生疤痕组织有可能转变成正常皮肤。     

人表皮干细胞的体外培养与鉴定

目的 探讨人表皮干细胞的体外分离培养方法 ,并对培养的表皮干细胞进行鉴定.方法 运用中性蛋白水解酶和胰蛋白酶两步法分离表皮层和真皮层,并获得表皮单细胞悬液,采用Ⅳ型胶原铺板选择性黏附分离角质形成细胞,应用表皮干细胞培养基进行培养.倒置显微镜下观察培养细胞的生长状况,免疫组织化学染色观察表皮干细胞β1整合素、角蛋白19(K19)、CK10的表达. 结果 组织学观察显示,培养中实验组细胞呈现克隆生长,且克隆维持时间较长;免疫组织化学染色显示,实验组的培养细胞β1整合素及K19均呈阳性表达,而CK10阴性表达.结论 应用Ⅳ型胶原黏附和运用表皮干细胞培养基可以实现人体表皮干细胞的分离和培养,为表皮干细胞体外的大量扩增奠定了基础.     

复合壳多糖人工皮肤的初步实验研究

目的探讨复合壳多糖人工皮肤移植家兔后表皮层角蛋白的表达以了解移植后皮肤表皮的分化情况.方法分别将复合壳多糖人工皮肤移植前、移植后1个月、移植后2个月、正常人皮肤、正常兔皮肤石蜡块切片,用抗广谱角蛋白多抗、抗角蛋白K5和K10单抗进行免疫组化染色,观察表皮层角蛋白的表达和表皮分化情况.结果复合壳多糖人工皮肤移植后1个月及2个月表皮层均有较强的角蛋白表达,与正常皮肤相似;基底层以上细胞有角蛋白K10表达,且越接近表皮表面表达越强,与正常人皮肤相似,基底层细胞有角蛋白K5表达,但明显弱于正常人皮肤.结论复合壳多糖人工皮肤移植家兔后表皮层均有角蛋白表达,有增生性细胞和分化细胞,表皮分化良好、层次清楚,与正常皮肤相近.     

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…