像素铒激光联合角质细胞生长因子在面部皮肤年轻化治疗中的应用

目的：探讨像素铒激光与角质细胞生长因子联合应用进行面部年轻化治疗的效果。方法：采用2940nm铒激光与角质细胞生长因子喷雾对68例患者进行治疗,共5次,每次间隔3周。结果：患者无不适反应,面部老化表现均得到较明显改善,随访6～12个月,患者的满意度均较高。结论：像素铒激光联合角质细胞生长因子喷雾对于面部年轻化的治疗其疗效较为持久,副作用小,操作简便,对于改善和延缓各种面部衰老的表现具有较好的效果。     

成纤维细胞生长因子的生物学功能及应用综述

成纤维细胞生长因子的生物学功能及应用综述呙长模＊张代录＊吴凯南＊成纤维细胞生长因子（ｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃ－ｔｏｒ，ＦＧＦ）是一类具广泛生物学活性的肽类物质，按其等电点（ＰＩ）的不同，可分为碱性成纤维细胞生长因子（ａｃｉｄｉｃｆｉｂｒ...     

角质细胞生长因子研究进展

目的 综述角质细胞生长因子(keratinocyte growth factor,KGF)的研究进展,全面了解KGF基本特性及应用方法 .为研制新型KGF药物、改善皮肤替代物的性能奠定基础. 方法 广泛查阅近年来国内外有关KGF的相关文献,并综合分析. 结果 KGF由各种来源的间质细胞分泌,受体分布于上皮细胞,特异性地促进上皮细胞增殖、迁移及分化,与器官发育、创面修复、肿瘤发生及免疫重建等多方面关系密切. 结论 KGF可用于促进创面愈合及改善皮肤替代物性能,但尚需改变结构,去除副作用,并纯化其促上皮生长的作用.     

角质形成细胞生长因子的研究进展

角质形成细胞生长因子（keratinocyte growth factor．KGF）是由间质细胞分泌的，通过旁分泌途径刺激上皮增殖的细胞因子。研究表明KGF在组织修复过程中发挥着很重要的作用，这些作用主要是KGF通过加强上皮屏障功能完成，包括促进细胞的增殖、迁移、分化、存活等。重组人角质形成细胞生长因子Palifermin己由FDA批准用于治疗放化疗导致的严重口腔粘膜炎。本文将对KGF的生物学特性、表达调控、生物学功能、及重组KGF的研究和应用进行讨论。     

角质形成细胞和成纤维细胞两步法培养的实验观察

目的应用中性蛋白酶和胰蛋白酶两步消化法原代培养角质形成细胞和成纤维细胞，并进行传代培养。方法取乳鼠背部皮肤，分离出表皮和真皮，加入中性蛋白酶和胰蛋白酶消化，进行原代和传代培养，通过倒置显微镜观察细胞的生长情况。结果原代培养的角质形成细胞和成纤维细胞长成单层且分布均匀；传代培养后排列规律、生长良好。结论两步消化法可以培养出角质形成细胞和成纤维细胞，并且比以往的培养方法更加方便快捷。     

角质形成细胞与成纤维细胞对中波紫外线照射应激能力的比较研究

目的：观察两种皮肤细胞即原代角质形成细胞及成纤维细胞对中波紫外线照射的反应差异。方法：采用不同剂量中波紫外线(UVB)照射上述两种细胞，评估UVB对细胞作用的时间及剂量效应，以倒置光学显微镜观察细胞受损程度，MTT法检测细胞活性。结果：UVB照射后；细胞损伤程度均随照射剂量加大而加重。另经24、48、72小时孵育后，分别对两种细胞进行不同时间段的细胞活性(MTT)比值比较(72／24小时，72／48小时)，发现细胞活性比依次递减：原代角质形成细胞(1．16和1．63)，成纤维细胞(1．05和1．09)。MTT结果显示低剂量紫外线照射时，细胞损伤恢复可发生在照射后48小时；高剂量紫外线照射后，细胞的损伤程度均随孵育时间延长而加重。结论：原代角质形成细胞抵抗紫外线辐射损伤的能力较强，而成纤维细胞相对较易受损。     

角质细胞生长因子和环氧化酶-2在胃癌中的表达及其与血管生成的关系

目的检测角质细胞生长因子(KGF)和环氧化酶-2(COX-2)蛋白在胃癌组织中的表达及微血管密度(MVD),探讨KGF与COX-2在胃癌发生、发展中的作用及其机理。方法采用免疫组化SP法检测64例胃癌组织及30例正常胃黏膜组织中KGF和COX-2蛋白表达,并采用CD34抗体染色检测MVD。结果 KGF和COX-2蛋白在胃癌组织中的表达阳性率分别为65.6%(42/64)和79.7%(51/64),分别高于其在正常胃黏膜组织中的表达阳性率〔23.3%(7/30)和13.3%(4/30)〕,P=0.046、P=0.008。胃癌组织MVD为31.8±8.0,明显高于正常胃黏膜组织的14.3±6.1(P=0.000);KGF与COX-2蛋白均表达阳性者MVD为35.9±5.7,明显高于两者表达均为阴性者的25.7±7.0(P=0.000)。胃癌组织中KGF和COX-2蛋白表达均与淋巴结转移、浆膜浸润及TNM分期有关(P<0.05、P<0.01),MVD与淋巴结转移和TNM分期有关(P<0.01),但均与患者年龄、性别以及肿瘤分化程度无关(P>0.05)。KGF与COX-2联合表达者与胃癌的浆膜浸润、淋巴结转移和TNM分期有关(P<0.05),与患者年龄、性别以及肿瘤分化程度无关(P>0.05)。胃癌组织中KGF与COX-2蛋白表达呈正相关(r=0.610,P=0.000);胃癌组织MVD与KGF和COX-2蛋白表达均呈正相关(r=0.675,P=0.000;r=0.657,P=0.000)。结论 KGF和COX-2蛋白在胃癌组织中高表达,可能通过促进肿瘤新生血管的生成协同参与胃癌的浸润、转移。     

碱性成纤维细胞生长因子对深Ⅱ°烫伤大鼠创面再上皮化过程中表皮干细胞的生物学作用

目的观察碱性成纤维细胞生长因子（bFGF）促进大鼠深Ⅱ°烫伤创面再上皮化作用及对表皮干细胞增殖和迁移的影响。方法66只Wistar大鼠随机分为A组（正常对照组，n=6）、B组（单纯烫伤组，n=30）、C组（bFGF治疗组，n=50）。利用大鼠5％深Ⅱ°烫伤模型，于伤后1、3、7、14和21d采取创面边缘皮肤标本，免疫组织化学染色技术检测创面边缘上皮及深层皮肤附件上皮整合素-β1（integrin-β1）、角蛋白19（K19）和PCNA以及上皮钙依赖性黏附素（E—cadherin）的表达情况。结果正常皮肤中，整合素-β1、角蛋白19、PCNA和E-cadherin表达主要集中在Wistar大鼠表皮层的基底部及毛囊球部。皮肤烫伤初期，以上指标变化不显著。7—14d时，整合素-β1和角蛋白19同时阳性表达的细胞有所增强，PCNA和E—cadherin的表达增强。当局部外用bFGF后，创伤初期（1—3d）组织中的表达差异无统计学意义（P〉0．05），7—14d，阳性表达角质形成细胞出现在创缘的棘层与颗粒层，毛囊根部的阳性标记细胞强于对照组。结论bFGF促进深Ⅱ°烫伤大鼠创面愈合与加速启动创缘表皮基底层的表皮干细胞以及皮肤附件上皮的迁移有关。     

角质细胞与成纤维细胞混合移植修复裸鼠全层皮肤缺损的初步报告

目的：利用组织工程原理探讨修复全层皮肤缺损的理想方式。方法：以裸鼠为动物模型,在皮肤全层缺损区域分别移植纤维蛋白胶(n=10),纤维蛋白胶角质细胞悬液（n=10）,纤维蛋白胶成纤维细胞悬液（n=10）以及纤维蛋白胶角质细胞成纤维细胞悬液（n=10）,术后每天对伤口进行大体观察,第5,7,10,14,21,35d,分别取材活检行组织学及免疫组织化学检查。结果：移植有角质细胞组（2和4组）的创面愈合快,术后10d组织学提示创面完全上皮化,抗人特异性HLA－1型抗原、抗involucrin染色和抗Ⅶ型胶原染色阳性证明新生上皮由移植的人角质细胞形成,抗involucrin染色阳性又证明角质细胞分化成熟有角质层形成,抗Laminin染色、抗Ⅶ型胶原染色阳性提示早期基底膜形成。组织学检查提示第4组新生上皮有许多类似皮钉样结构。结论：培养的角质细胞,成纤维细胞结合纤维蛋白胶移植到创面上后,可以形成复层分化良好、接近正常结构和功能的新生成肤组织。     

酪氨酸酶活性抑制实验及其在祛斑美白化妆品功效评价中的应用

目的：建立评价祛斑美白化妆品功效的体外实验方法.方法：根据酪氨酸酶催化底物发生反应的原理，研究建立和优化了一种酪氨酸酶活性抑制实验系统，并采用不同浓度的曲酸溶液作为标准对照.结果：应用本方法进行测试，分别得出3.0%的熊果苷、4.0%的曲酸双棕榈酸脂、0.5%的氢醌对酪氨酸酶活性的抑制率分别为58.21%、92.92%、126.01%.对未知成分的数种化妆品样品也进行了同样测试，抑制率在3.1 5%～47.69%.结论：本研究为祛斑美白化妆品的功效评价提供了一种体外实验方法。     

生长分化因子-9在人膀胱癌组织中的表达及其临床意义

目的：探讨生长分化因子-9（growth differentiation factor-9,GDF-9）在人膀胱癌组织中的表达及其与膀胱癌病理分级和临床分期的关系。方法：运用免疫组化技术检测GDF-9在50例不同分期分级的膀胱癌组织和15例正常膀胱组织之间的蛋白表达情况。结果：①正常人膀胱组织中GDF-9呈高表达,且GDF-9蛋白主要定位于膀胱移行上皮细胞的胞浆中。而膀胱癌组织中GDF-9呈低表达或不表达。②GDF-9阳性表达率在膀胱癌病理分级间分别：I级47．6％（10／21）,II级26．7％（4／15）,III级7．1％（1／14）,差异具有统计学意义（P〈0．05）。GDF9阳性表达率在膀胱癌临床分期间分别：Ta～T1期40．6％（13／32）,T2～T3期11．1％（2／18）,差异具有统计学意义（P％0．05）。结论：GDF-9在膀胱癌组织中低表达或不表达,且与膀胱癌病理分级和临床分期呈负相关。GDF-9可能是一个潜在的人膀胱肿瘤抑癌基因。     

转移生长因子—β,碱性成纤维细胞生长因子诱导人成骨细胞表达血小板衍生生

目的 探讨转移生长因子（ＴＧＦ）－β、碱性成纤维细胞生长因子（ｂＦＧＦ）对人成骨细胞血小板衍生生长因子（ＰＤＧＦ）－ＢｍＲＮＡ表达的影响及其意义。方法 体外分离培养人成骨细胞。在体外培养的成骨细胞中分别加入１０、２０、４０、８０、１６０ｕｇ／Ｌ梯度浓度的ＰＤＧＦ－ＢＢ培养细胞２４ｈ,以摄取＾３Ｈ－ＴｄＲ为细胞增殖指标检测细胞增殖状况。以４ｕｇ／Ｌ的ＴＧＦ－β和１０ｕｇ／Ｌ的ｂＦＧＦ培养细胞２４ｈ,寡核苷酸探针检测细胞ＰＤＧＦ－ＢｍＲＮＡ的表达。结果 １０～１６０ｕｇ／Ｌ的ＰＤＧＦ－ＢＢ可促进成骨细胞增殖（Ｐ〈０．０５）。在普通培养条件下,细胞下表达ＰＤＧＦ－ＢｍＲＮＡ;当培养体系中加入ＴＧＦ－β和ｂＦＧＦ,可见ＰＤＧＦ－ＢｍＲＮＡ的表达。结论 ＰＤＧＦ－Ｂ基因的表达可能是骨组织生长的储备因素,ＴＧＦ－β和ｂＲＮ     

Role of basic fibroblast growth factor-2 in epithelial-mesenchymal transformation

BACKGROUND: Epithelial-mesenchymal transformation (EMT) plays an important role in embryonic development and tumorigenesis and has been described in organ remodeling during fibrogenesis. In the kidney, EMT can be induced efficiently in cultured proximal tubular epithelium by coincubation of transforming growth factor (TGF)-beta1 and epidermal growth factor (EGF). Recently, we also have observed overexpression of basic fibroblast growth factor-2 (FGF-2) protein and mRNA in human kidneys with marked interstitial fibrosis. The aims of the present study were to compare the effects of FGF-2 as a facilitator of EMT in tubular epithelial cells with EGF and TGF-beta1. We analyzed the morphogenic effects of the three cytokines on four different aspects of EMT: cell motility, expression and regulation of cellular markers, synthesis and secretion of extracellular matrix (ECM) proteins as well as matrix degradation. METHODS: Cell motility was studied by a migration assay and cell differentiation markers were analyzed by immunofluorescence and immunoblots. In addition, regulation of the epithelial adhesion molecule E-cadherin and fibroblast-specific protein 1 (FSP1) were analyzed by luciferase reporter constructs and stable transfections. ELISAs for collagen types I and IV and fibronectin were used for ECM synthesis, and zymograms were utilized for analysis of matrix degradation. RESULTS: FGF-2 induced cell motility across a tubular basement membrane in two tubular cell lines. All three cytokines induced the expression of vimentin and FSP1, but only FGF-2 and TGF-beta1 reduced cytokeratin expression by immunofluorescence. These effects were most demonstrable in the distal tubular epithelial cell line and were confirmed by immunoblot analyses. Expression of E-cadherin was reduced by 61.5 +/- 3.3% and expression of cytokeratin by 91 +/- 0.5% by TGF-beta1 plus FGF-2. Conversely, the mesenchymal markers alpha-smooth muscle actin (SMA) and FSP1 were induced with FGF-2 by 2.2 +/- 0.1-fold and 6.8 +/- 0.9-fold, respectively. Interestingly, de novo expression of the mesenchymal marker OB-cadherin was induced only by FGF-2 and EGF but not by TGF-beta1. All three cytokines stimulated FSP1 and decreased E-cadherin promoter activity. FGF-2 also induced intracellular fibronectin synthesis but not secretion, the latter of which was stimulated exclusively by TGF-beta1. Finally, zymographic analyses demonstrated that FGF-2 induced MMP-2 activity by 2.6 +/- 0.5-fold and MMP-9 activity by 2.4 +/- 0.1-fold, providing a mechanism for basement membrane disintegration and migratory access of transforming epithelium to the interstitium. CONCLUSIONS: FGF-2 makes an important contribution to the mechanisms of EMT by stimulating microenvironmental proteases essential for disaggregation of organ-based epithelial units. Furthermore, the expression of epithelial and mesenchymal marker proteins seems to be affected at the promoter level.     

碱性成纤维细胞生长因子在大鼠神经病理性痛中的作用

目的 评价碱性成纤维细胞生长因子(FGF-2)在大鼠神经病理性痛中的作用.方法 雄性SD大鼠100只,7周龄,体重200 ～ 250 g,采用随机数字表法,将大鼠随机分为4组(n=25):假手术组(S组)、神经病理性痛组(NP组)、PBS组、FGF-2抗体组(Ab组).于术后1、6、9、13、16、20d时Ab组鞘内注射FGF-2抗体18μg(40 μl),PBS组鞘内注射等容量的PBS溶液.采用坐骨神经分支选择性损伤的方法制备神经病理性痛模型.于术前ld,术后1、4、7、14、21 d时观察痛行为学,测定机械缩足反射阈值(PWMT),并于术后各时点应用ELISA法检测脊髓TNF-α和IL-6含量.结果 与S组比较,NP组、PBS组和Ab组PWMT降低,脊髓TNF-α和IL-6含量升高(P＜0.05).与NP组和PBS组比较,Ab组PWMT升高,脊髓TNF-α和IL-6含量降低(P＜0.05).结论 FGF-2参与大鼠神经病理性痛的发生和发展,其机制与诱发脊髓组织炎性反应有关.     

Neutralization of fibroblast growth factor-2 reduces intraarticular adhesions

Adhesion is a serious complication after trauma or surgery. Because adhesion formation is essentially a fibrogenetic process, a series of growth factors are assumed to be involved in its development. If this is true, it may be possible that inhibition of the growth factor activity suppresses adhesion formation. The current study was conducted to verify this hypothesis on fibroblast growth factor-2 using an intraarticular adhesion model in the rabbit knee. Forty Japanese White rabbits were used. They were divided randomly into five groups of eight animals, and in three of them, activity of endogenous fibroblast growth factor-2 was suppressed locally by a neutralizing antibody. The remaining two groups served as controls, and formation of adhesions was evaluated 4 weeks after surgery. The results showed that the administration of the antibody reduced the extent of adhesions macroscopically, whereas histologic observation and collagen content measurement suggested the adhesion tissue was not affected significantly. Corresponding to the macroscopic findings, contraction of the knee was improved in the antibody groups. The findings showed that suppression of fibroblast growth factor-2 activity reduces adhesions. It is expected that control of the cytokine activity may become a novel method for reducing adhesions.     

Isoproterenol inhibits fibroblast growth factor-2-induced growth of renal epithelial cells

The signal transduction pathways modulating bFGF effects in renal tubular epithelial cells (RTEc) are not completely understood. Since the cAMP and the mitogen-activated protein kinase (MAPK) pathways can modulate the growth of RTEc, we studied whether two cAMP elevating agents, isoproterenol and 8-bromo-cAMP, would modulate basic fibroblast growth factor (bFGF) induction of MAPK activity (ERK-2) and cell proliferation in human renal proximal tubular epithelial cells (RPTEc) and Madin-Darby canine kidney cells (MDCK clone _EI1). Isoproterenol, but not bFGF, stimulated cAMP production in RPTEc and MDCK_EI1 cells. bFGF, isoproterenol, and 8-bromo-cAMP alone increased ERK-2 activity in both cell types. However, isoproterenol and 8-bromo-cAMP partially inhibited the bFGF induction of ERK-2 activity, but only isoproterenol inhibited the proliferation of both cell types. PD098059 (25 μM), an inhibitor of MAPK kinase (MEK 1/2), blocked the bFGF mitogenic effects, but did not affect the 8-bromo-cAMP-induced mitogenic effects in MDCK_EI1 cells. These findings suggest that activation of ERK-2 is required but not sufficient for mitogenesis in RTEc. We conclude that isoproterenol inhibits the growth-promoting effects of bFGF in RTEc via MEK-dependent and -independent pathways. Received: 15 April 1999 / Revised: 9 July 1999 / Accepted: 10 August 1999     

化妆品中丙烯酰胺及其检测的研究进展

丙烯酰胺是生产聚丙烯酰胺的原料,在化妆品中存在的微量丙烯酰胺是生产过程中添加聚丙烯酰胺引入的。由于丙烯酰胺具有神经毒性和潜在致癌性,已被列为化妆品中的限制使用成分,加强化妆品中丙烯酰胺的监测具有重要意义。本文综述了丙烯酰胺的来源、用途和危害性,总结了目前化妆品中丙烯酰胺测定的所用的样品前处理方法,以及丙烯酰胺气相色谱法、液相色谱法、液相色谱-串联质谱法等各种检测方法的最新研究进展,同时对化妆品样品中丙烯酰胺检测技术的发展趋势进行了展望,表明采用串联质谱技术（如GC-MS/MS、LC-MS/MS）对进行复杂样品中丙烯酰胺进行检测是未来的发展趋势。     

Effects of high-flux hemodialysis on fibroblast growth factor-23 and its clinical significance

Objective To investigate the effects of high-flux hemodialysis (HFD) on fibroblast growth factor-23 (FGF-23) levels in maintenance hemodialysis (MHD) patients and its clinical significance. Methods Sixty uremia patients were divided into HFD group and hemodialysis (HD)group and observed for 12 months. Flow mediated dilation (FMD), cardiac ultrasonography, levels of FGF-23, serum phosphorus, serum calcium, 25-(OH)D3, parathyroid hormone (PTH), homocysteine(Hcy) and interleukin-6 (IL-6) were tested in all patients before and after treatment. The correlation ofabove indexes were analyzed. Results No statistical difference were found in primary disease, age and duration of dialysis in two groups. The levels of FGF-23 [(56.07±26.63) vs (85.53±40.54) ng/L, P＜0.01], IL-6 [(3.37±2.48) vs (5.59±2.53) ng/L, P＜0.05] and Hcy [(21.13±6.95) vs (29.40±11.66) μmol/L, P＜0.05] decreased and FMD, 25-(OH)D3 [(27.3±10.26) vs (23.15±10.73) μg/L, P＜0.05] increased significantly after the treatment of HFD. There were no significant changes in the HD group. The baseline FMD was negatively correlated with FGF-23 (r＝-0.413, P＜0.05) and Hcy (r＝-0.301, P＜0.05). The baseline LVMI was correlated with FGF-23 (r＝0.464 P＜0.05). After one year's trearmeat of HFD, the changes of FMD(△FMD) was negatively correlated with the changes ofFGF-23 (△FGF-23)(r＝-0.347, P＜0.05). Conclusions HFD can improve FMD and decrease FGF-23 levels. The improvement of FMD may be related to the decreased level of FGF-23. The effect of FGF-23 on FMD should be independent of serum phosphate.