Caspase-3／Caspase-9活化与皮鳞癌1号方抑制A431细胞生长的关系研究

目的：研究 Caspase-3和 Caspase-9活化与皮鳞癌1号方抑制 A431细胞生长的关系。方法培养 A431细胞,用不同浓度的皮鳞癌1号方分别作用于 A431细胞;Hochest 染色检测细胞凋亡形态;酶联免疫吸附测定法（ELISA）检测细胞色素 C 含量;比色法测定 Caspase-3和 Caspase-9活性。结果Hochest33258染色可见皮鳞癌1号方组 A431细胞发生核固缩、凋亡小体等典型的凋亡形态学特征;与对照组比较,Cytc 、Caspase-3、Caspase-9表达增加。结论Caspase-3和 Caspase-9的活化与皮鳞癌1号方可诱导 A431发生凋亡有关。     

黄芪甲苷Ⅳ联合紫杉醇通过STAT3-NF-κB途径对胃癌细胞的作用研究

目的探讨黄芪甲苷Ⅳ联合紫杉醇对胃癌细胞的作用及其可能的机制。方法黄芪甲苷Ⅳ和紫杉醇按给药剂量分为0μg/ml组、5μg/ml组、10μg/ml组、20μg/ml组、40μg/ml组、80μg/ml组和160μg/ml组处理胃癌细胞AGS。CCK-8试剂盒检测细胞活力;集落形成实验检测细胞增殖;流式细胞术检测细胞凋亡;Western blot检测Bax、Bcl-2和STAT3-NF-κB途径相关蛋白的表达;Transwell实验检测细胞迁移和侵袭。结果黄芪甲苷Ⅳ在20μg/ml、40μg/ml、80μg/ml和160μg/ml浓度下可明显抑制AGS细胞活力,黄芪甲苷Ⅳ IC_(50)=63.99μg/ml;紫杉醇在10μg/ml、20μg/ml、40μg/ml、80μg/ml和160μg/ml浓度下可明显抑制AGS细胞活力,紫杉醇IC_(50)=40.08μg/ml。与对照组相比,黄芪甲苷Ⅳ组、紫杉醇组和联合组的细胞活力、克隆形成数、细胞迁移和侵袭数明显降低(P<0.05),细胞凋亡率、Bax蛋白表达明显升高(P<0.05),Bcl-2、pSTAT3/STAT3和pNF-κB/NF-κB蛋白表达明显降低(P<0.05);与联合组比较,黄芪甲苷Ⅳ组和紫杉醇组的细胞活力、克隆形成数、细胞迁移和侵袭数明显升高(P<0.05),细胞凋亡率、Bax蛋白表达明显降低(P<0.05),Bcl-2、pSTAT3/STAT3和pNF-κB/NF-κB蛋白表达明显升高(P<0.05)。结论黄芪甲苷Ⅳ联合紫杉醇可抑制胃癌细胞增殖、迁移和侵袭,促进细胞凋亡,该作用可能是通过抑制STAT3-NF-κB途径实现的。     

葛根素对人小细胞肺癌H446细胞增殖与凋亡的影响及其作用机制研究

目的观察葛根素对人小细胞肺癌H446细胞增殖与凋亡的影响,并初步探讨其作用机制。方法采用MTr比色法和流式细胞术检测葛根素对H446细胞增殖抑制和诱导凋亡的作用;免疫组化法检测葛根素对PCNA、Bax和Bcl-2蛋白表达的影响。结果40、80μg/ml浓度的葛根素体外作用24、48h,对H446细胞有促增殖作用,在160—640μg／ml浓度范围内,葛根素能够抑制H446细胞的生长,并呈现良好的剂量依赖和时间依赖关系。435μg/ml的葛根素可以诱导H446细胞凋亡。检测葛根素对PCNA、Bax和Bcl-2蛋白表达的影响显示葛根素能够下调PCNA和Bcl-2蛋白的表达,上调Bax蛋白的表达。结论在一定浓度范围内,葛根素可能通过下调PCNA和Bcl-2蛋白的表达,上调Bax蛋白的表达而诱导H446细胞凋亡。     

锰超氧化物歧化酶模拟化合物对人胃癌MGC-803细胞增殖和凋亡的影响

目的观察Mn SOD模拟化合物(Mn SODm)对人胃癌MGC-803细胞增殖和凋亡的影响。方法体外培养人胃癌MGC-803细胞,MTT法观察Mn SODm对MGC-803细胞增殖的影响;用Hoechst33258染色观察MGC-803细胞形态学的变化;Annexin V-FITC/PI检测MGC-803细胞凋亡;Western blot法检测p53、cleaved caspase-3、cleaved caspase-9、Bcl-2、Bax蛋白的表达。结果 MTT法检测结果显示1~20μg·m L~(-1)Mn SODm对MGC-803细胞有显著的抑制作用,24、48、72 h的IC50分别为10.18、6.93和5.05μg·m L~(-1);20μg·m L~(-1)Mn SODm作用细胞48 h后,细胞凋亡率为(69.33±4.07)%(P<0.01);Western blot结果显示,用5、10、20μg·m L~(-1) Mn SODm处理细胞48 h后,Bcl-2表达显著降低,同时p53、cleaved caspase-3、cleaved caspase-9和Bax表达显著增加(P<0.05或P<0.01)。结论 Mn SODm对人胃癌MGC-803细胞有明显的抑制作用,可能是通过下调Bcl-2表达,增加p53、cleaved caspase-3、cleaved caspase-9和Bax表达来诱导MGC-803细胞凋亡。     

红三叶异黄酮对人宫颈癌HeLa细胞的抑制作用

目的探讨红三叶异黄酮对人宫颈癌HeLa细胞的抑制作用及其作用机制。方法以MTT法检测红三叶异黄酮20、40、80μg/m L分别在24、48、72 h对人宫颈癌HeLa细胞增殖作用的影响,并在倒置显微镜下观察HeLa细胞形态学改变。流式细胞仪检测红三叶异黄酮20、40、80μg/m L对HeLa细胞凋亡的影响。实时荧光定量PCR仪检测红三叶异黄酮20、40、80μg/m L对HeLa细胞凋亡中调节基因Bax、Bcl-2表达水平的影响。结果红三叶异黄酮对人宫颈癌HeLa细胞的增殖抑制作用,表现为时间、浓度的相关性。红三叶异黄酮对HeLa细胞的形态学改变,表现为随药物浓度增加细胞固缩变小,并出现凋亡小体。流式细胞仪检测分析显示红三叶异黄酮能引起HeLa细胞凋亡,并随药物浓度的增加而明显,当红三叶异黄酮达到一定有效浓度时,诱导其细胞凋亡。通过调节Bax mRNA与Bcl-2 mRNA的表达,促进抑制HeLa细胞增殖,能使Bax表达上升、Bcl-2表达下降,且呈剂量相关性。结论红三叶异黄酮对人宫颈癌HeLa细胞的增殖具有抑制作用,并呈一定的剂量和时间相关性,提示通过调节Bax mRNA与Bcl-2 mRNA的表达,促进抑制HeLa细胞增殖。     

异补骨脂素对L02细胞增殖、周期和凋亡的影响

目的研究异补骨脂素对人正常肝细胞(L02)增殖、细胞周期和凋亡的影响,并探讨其相关作用机制。方法以异补骨脂素为研究对象,L02细胞为体外模型,采用MTT试验检测不同浓度异补骨脂素(5、10、20、40、80和100μg/ml)对L02细胞增殖的影响,流式细胞仪检测不同浓度异补骨脂素体外作用24 h对L02细胞周期分布和凋亡率的影响,通过蛋白印迹法(Western blot)检测细胞周期蛋白D1(Cyclin D1)、细胞周期蛋白E1(Cyclin E1)、细胞周期素依赖性激酶2(CDK2)、细胞周期素依赖性激酶4(CDK4)、B淋巴细胞瘤-2基因(B-cell lymphoma 2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2-like protein4/Bcl-2-associated X protein,Bax)的表达,探讨异补骨脂素毒性的作用机制。结果 MTT试验表明,异补骨脂素对L02细胞生长有明显抑制作用,且抑制作用呈现时间-剂量依赖性;流式细胞仪检测结果显示,各组细胞经异补骨脂素(20、40和80μg/ml)作用24 h后,细胞总凋亡率分别为5.59%,7.8%和20.91%(P0.001),40和80μg/ml异补骨脂素组L02细胞处于G1期的细胞数增多,S期的细胞减少,说明异补骨脂素能将L02细胞的细胞周期阻滞在G1期;Western blot蛋白质印迹结果显示,同对照组相比,不同浓度异补骨脂素处理L02细胞24 h后,Cyclin D1蛋白表达水平明显下降(P0.05),而Bax蛋白表达水平明显升高,差异有统计学意义(P0.05)。结论异补骨脂素可能通过调控周期蛋白及细胞凋亡相关蛋白抑制L02细胞增殖,并诱导细胞凋亡。     

二烯丙基二硫对Raji细胞化疗增敏的研究

目的研究二烯丙基二硫（dimlyldisulfide,DADS）对长春新碱（VCR）作用于人淋巴瘤Raji细胞化疗增敏的作用及机制。方法DADS（0．625μg／ml、1．25μg／ml）及VCR（6．25,12．5,25．0μg／ml）单用及联合应用作用于Raji细胞,采用CCK-8（cellcountingkit）法检测各自的抑制率,应用金氏公式求q值。评价联合用药效果;DADS（0．625μg／ml）、VCR（6．25,12．5,25．0μg／ml）及联合应用作用于Raii细胞,采用流失细胞术检测细胞凋亡率,采用细胞免疫化学法检测细胞Bax及Bcl-2的表达情况。结果无毒剂量的DADSfo．6251Ag／ml、1．25μg／ml）分别与VCR联合应用,可提高VCR对Raji细胞增殖抑制率。DADS（0．625μg／m1）提高VCR（6．25,12．5,25．0μg/ml）诱导Raji细胞凋亡率（P〈0．05）。联合组Raji细胞Bcl-2蛋白表达水平比VCR单药组低（P〈0．05）;Bax蛋白表达水平在联合组Raji细胞VCR单药组高（P〈0．05）。结论DADS能增强VCR对Raji细胞增殖抑制和诱导凋亡作用．起化疗增敏作用：可能机制与其下调Bcl-2蛋白表达水平,上调Bax蛋白表达水平有关。     

色素上皮衍生因子在A431细胞的表达及对其增殖的影响

目的明确色素上皮衍生因子（pigment epithelium-derived factor, PEDF）在A431细胞的表达情况以及对其增殖影响。方法采用RT-PCR和Western—blot分别检测A431细胞PEDFmRNA和蛋白的表达：MTT方法检测PEDF对A431细胞增殖的影响。结果A431细胞在mRNA和蛋白水平均可检测到PEDF的表达。此外,100gg/ml的PEDF可以抑制A431细胞的增殖。结论PEDF抑制A431细胞增殖,有望用于皮肤鳞癌的治疗。     

复方绞股蓝胶囊对口腔鳞癌CAL27细胞增殖和表达的影响

目的研究复方绞股蓝胶囊（CoFG）对口腔鳞癌CAL27细胞MMP-2,9和Bax、Bcl-2表达的影响。方法采用MTT法检测CoFG对口腔鳞癌CAL27细胞增殖抑制的作用;qPCR法和Western blot分别检测CoFG对口腔鳞癌CAL27细胞MMP-2,9和Bax、Bcl-2 mRNA及蛋白表达的影响。结果 CoFG组和非CoFG组在细胞增殖抑制方面差异有统计学意义;CoFG下调MMP-2、Bcl-2 mRNA及蛋白表达;CoFG上调MMP-9、Bax mRNA及蛋白表达。结论 MMP-2,9和Bax、Bcl-2在口腔鳞癌的发生发展中起一定作用,CoFG可能对口腔鳞癌的治疗具有一定的潜在作用。     

尿石素A对乳腺癌细胞MCF-7增殖、凋亡的影响及其机制研究

目的 研究尿石素A对乳腺癌细胞MCF-7增殖、凋亡的影响并探讨其作用机制。方法 CCK-8法考察不同浓度尿石素A作用12、24、36、48 h对MCF-7细胞增殖能力的影响;细胞凋亡染色法考察20、40 μmol/L的尿石素A对MCF-7细胞凋亡的影响;实时荧光定量PCR（RT-qPCR）检测c-Myc、Cyclin D1、Bcl-2、Bax mRNA的表达水平;Western blotting法检测c-Myc、Cyclin D1、Bcl-2、Bax蛋白的表达水平。结果 尿石素A对MCF-7细胞增殖具有抑制作用且呈时间浓度相关性;细胞凋亡染色显示,20、40 μmol/L尿石素A给药后均能够诱导MCF-7细胞凋亡;RT-qPCR及Western blotting结果显示,20、40 μmol/L尿石素A能够显著降低MCF-7细胞中c-Myc、Cyclin D1、Bcl-2 mRNA及蛋白的表达水平（P<0.05、0.01）,升高Bax mRNA及蛋白的表达水平（P<0.05、0.01）。结论 尿石素A具有抑制MCF-7细胞增殖并诱导其凋亡的作用,其作用机制可能与抑制c-Myc、Cyclin D1、Bcl-2表达,升高Bax表达水平有关。     

Analysis of Drug Penetration Through the Skin by the Two-Layer Skin Mode

A diffusion model for the skin penetration of drug in the finite-dose system was developed considering the skin to be composed of two layers, the outermost layer (stratum corneum) and the lower layer (viable epidermis and dermis). Based on this skin model, the Laplace transforms of the equations for the drug amounts in the receptor, the vehicle, and the skin were derived. The penetration profiles of 6-mercaptopurine (6-MP) through the intact and stripped guinea pig skin were obtained from in vitro diffusion experiments. The computer fitting of those profiles to the Laplace-transformed equations by a nonlinear least-squares program based on a fast inverse Laplace transform algorithm [MULTI-(FILT)] gave parameters such as diffusion coefficients of 6-MP and thicknesses of both layers. The mean transit time (MTT) for each diffusion process was defined based on statistical moment concept and calculated using the obtained parameters. Under the present condition, the process to move from the vehicle to the stratum corneum is demonstrated to have the longest mean time in overall processes of 6-MP penetration.     

Metals and the Skin

Abstract

Certain metals, and many metal-based compounds, are inherently toxic, and their presence in occupational and environmental settings raises appropriate questions concerning human exposure. Contact of these materials with the skin represents an important route of exposure, which is not well characterized. The purpose of this review, therefore, is to assemble the available, useful information pertinent to risk assessment following dermal contact.

Specifically, we summarize here: (1) data relevant to the qualitative and (where possible) quantitative evaluation of metal compound permeation through the skin; (2) the role of each metal in metabolism, particularly with respect to the skin, and the potentially toxic effects that may result from dermal contact; and (3) the immunological characteristics (including allergenicity) of the metals and their derivatives.

In total, information on 31 metals has been reviewed. It is clear that many diverse factors determine the ability of metal-based species to permeate biological membranes, not all of which have been fully defined. Therefore, considerably more experimentation, targeted at the development of high-quality transport data, will be required before the specification of practically useful structure-activity relationships are possible.     

自体微粒-异体筛状复合皮供皮区的移植

目的提高供皮区愈合质量;方法应用鼓式取皮机切取人体不同部位中厚或厚中厚皮肤,采取配对分组法对观察组和对照组分别施以不同覆盖方法,根据组织学、肉眼观察和患者白评等方法对疗效进行判断,使用SAS统计软件包和配对等级分组资料的秩和检验法对资料进行统计处理;结果观察组和对照组在愈合时间上无明显差别,搔痒、刺痛和瘢痕程度等方面观察组优于时照组;结论白体微粒一异体筛状复合皮移植于供皮区有助于提供供皮区愈合质量。     

中厚皮片移植在局部晚期乳腺癌胸壁皮肤缺损中的应用

目的 探讨应用中厚游离皮瓣治疗局部晚期乳腺癌术后胸壁皮肤缺损的可行性.方法 回顾性分析我院自2011年以来收治的15例局部晚期乳腺癌病人的临床资料,患者在接受标准的乳腺癌改良根治术后,采用游离中厚皮片移植的方法修复胸壁的巨大皮肤缺损.结果 13例患者植皮区游离皮片完全成活,1例约20%的植皮区域出现表皮坏死,1例约10%的皮片坏死,经换药后痊愈.无围手术期死亡.结论 对于可手术的局部晚期乳腺癌患者,游离中厚皮片可以覆盖肿瘤切除后的胸壁皮肤缺损,具有较好的疗效.     

Sulphur Mustard Injuries of the Skin

Sulphur mustard is a vesicant (blistering agent), which produces chemical burns with widespread blistering. It was used extensively as a chemical warfare agent in the First World War, and has allegedly been employed in a number of conflicts since then, most recently by Iraq against Iran (1984–1987). The potential further use of mustard in military conflicts and by terrorists remains a significant threat that if realised in practice would result in a large number of casualties with severely incapacitating, partial thickness burns. Such injuries clearly present a huge potential wound care problem. The development and healing of mustard-induced cutaneous injuries has not only been observed in human casualties, but has been studied recently at the microscopic and ultrastructural levels in several animal models. Vesication generally begins on the second day after exposure, and may progress for up to 2 weeks. Wound healing is considerably slower than for a comparable thermal burn, and patients often require extended hospital treatment. The current management strategy is essentially symptomatic and supportive. Recently, two techniques for removing damaged tissue and improving wound healing have been investigated. Mechanical dermabrasion and laser debridement (‘lasablation’) both produced an increased rate of wound healing in animal models, and may be of benefit in a clinical context.     

Skin cancer in the elderly

With the significant increase in the average life-span in the industrial world, skin cancer has become a great health concern. There are various epidemiological, biological and molecular data suggesting that skin cancer is predominantly a disease of the elderly, since approximately 53% of skin cancer-related deaths occur in persons more than 65 years old. With regard to the management of elderly patients with skin cancer, this should be individualized depending upon the clinical performance status, and age alone should not constitute an obstruction for the administration of the optimal treatment. Since elderly patients with melanoma have a worse prognosis, emphasis should be given to primary and secondary prevention. Physicians treating elderly patients should be trained in an individualized approach to these patients and encouraged to participate in programs for the early detection of suspicious skin lesions.