猪脱细胞真皮基质对小鼠创面毛囊再生中基质细胞衍生因子-1及Wnt3a/β-catenin信号通路表达的影响

目的探讨猪脱细胞真皮基质(ADM)对小鼠创面毛囊再生中基质细胞衍生因子-1(SDF-1)及Wnt3a/β-catenin信号通路表达的影响。 方法取新鲜猪皮,经脱细胞处理后制成微粒状,灭菌,密封,常温保存,以备后面实验使用。制作18只C57BL/6小鼠背部全层皮肤缺损模型,以脊柱为中线,在左右各制作直径为6 mm的缺损,左右侧分别以纱布和猪ADM覆盖,隔天纱布侧换药,猪ADM侧不做处理,于模型建立第7天,按照所用材料不同分为纱布组与猪ADM组,再按所取部位不同分为纱布窗口组、猪ADM窗口组、纱布创面组、猪ADM创面组、纱布创缘组、猪ADM创缘组,其中9只小鼠组织用于蛋白质印迹法检测,另外9只小鼠组织用于免疫组织化学检测。通过蛋白质印迹法检测肿瘤坏死因子-α(TNF-α)、β-catenin、转化生长因子-β(TGF-β)、Wnt3a、血管内皮生长因子(VEGF)、SDF-1、成纤维细胞生长因子(FGF)2、FGF9、AKT的蛋白表达,免疫组织化学检测Wnt3a、SDF-1、FGF2、FGF9的表达。数据比较采用独立样本t检验。 结果在小鼠模型建立第7天创面中,蛋白质印迹法检测β-catenin、Wnt3a、SDF-1的蛋白表达量均是猪ADM窗口组(0.533±0.058、0.446±0.039、0.972±0.048)高于纱布窗口组(0.401±0.005、0.132±0.022、0.175±0.036),差异均有统计学意义(t＝3.996、12.230、23.130,P值均小于0.05)。β-catenin、Wnt3a、SDF-1的蛋白表达量均是猪ADM创面组(0.557±0.009、0.626±0.066、0.868±0.102)高于纱布创面组(0.302±0.010、0.109±0.019、0.036±0.009),差异均有统计学意义(t＝32.830、13.020、14.130,P值均小于0.05)。Wnt3a、SDF-1的蛋白表达量均是猪ADM创缘组(0.419±0.014、0.370±0.069)高于纱布创缘组(0.115±0.020、0.056±0.007),差异均有统计学意义(t＝21.460、7.825,P值均小于0.05)。免疫组织化学检测结果显示Wnt3a、SDF-1、FGF2、FGF9在猪ADM组中的表达高于纱布组,且阳性细胞主要分布于毛囊细胞周围。 结论猪ADM在小鼠创面中可能通过上调SDF-1及Wnt3a/β-catenin信号通路的表达而促进毛囊的再生。     

马桑提取物促进大鼠烧伤创面愈合的作用和机制

 目的：探讨马桑提取物(CSME)对大鼠烧伤创面愈合的影响及机制。方法：随机将40只SD雄性大鼠分成生理盐水（NS）组、白凡士林（WPJ）组、磺胺嘧啶银（SSD）组和CSME组,每组10只。将大鼠麻醉后,制备背部深Ⅱ度皮肤烧伤创面。而后分别用NS敷料、WPJ敷料、SSD敷料和CSME敷料覆盖治疗21 d。在第1 d、3 d、7 d、14 d和21 d,观察动物的临床症状和创面状况（上皮化速率、结痂及被毛等）后,分别取出创面组织,用于组织学观察,检测丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性,检测表皮生长因子（EGF）和碱性成纤维细胞因子（bFGF）蛋白水平及胶原mRNA表达。结果：CSME组新生上皮生长高度活跃,有大量新生被毛生长,创面上皮化速率明显高于其它各组 (P<0.05)。SSD组创面中央新生被毛稀疏而细小,而CSME组创面中央被毛密集,大部分被毛接近正常。CSME组镜下可见全部被多层上皮细胞覆盖,新生胶原纤维充分分化,排列整齐,组织结构明朗,皮脂腺和毛囊增生极其活跃等,显著优于其它各组。本研究显示,烧伤后从第1 d到21 d,CSME组早期EGF和bFGF水平显著高于其它各组,后期迅速下降,低于其它组（P<0.05或P<0.01）。SSD组Ⅰ型与Ⅲ型胶原mRNA表达的比值显著高于其它各组和正常组织,CSME治疗组显著低于其它各组和正常组织(P<0. 05)。结论：本研究表明,CSME可诱导烧伤创面无瘢痕愈合。CSME的这种作用与其早期增强EGF和bFGF蛋白水平,后期抑制两者的水平有关,也可能与其抑制烧伤创面Ⅰ型胶原而增强Ⅲ型胶原mRNA表达密切相关。     

异体脱细胞真皮基质对大鼠皮肤瘢痕形成的影响

目的了解异体脱细胞真皮基质（accelular dermal matrix,ADM）对SD大鼠皮肤切口愈合后瘢痕形成的影响。方法4％NaOH溶液消蚀脱细胞法制备ADM,选用SD大鼠56只,每只大鼠背部脊柱两侧共做两道平行切口,建立皮肤切口模型。按自身对照分为实验组及对照组,实验组：将修剪成的ADM（3cm×0.3cm×0．1cm）植入切口,丝线缝合切口;对照组则原位缝合。术后3天、5天、7天、14天、21天、28天、63天（每个时相点8只）处死大鼠,取切口皮肤组织做HE、Masson染色,进行生物组织学检测,RT-PCR方法检测各组切口中成纤维细胞内Ⅰ型前胶原mRNA的表达水平。结果术后实验组未见明显瘢痕组织,而对照组愈合后见明显线性瘢痕形成。镜下观察实验组切口中ADM逐渐被改建成自体组织,3天、5天、7天、14天及21天实验组中成纤维细胞数量较对照组减少（P〈0．05）;实验组切口中胶原的生成在各个时间点较对照组均减少,其中7天、14天、21天、28天及63天实验组切口中胶原的生成较对照组明显减少（P〈0．01）;RT-PCR检测组织中成纤维细胞内Ⅰ型前胶原mRNA相对表达量对照组（0．809±0．042）比实验组（0．540±0．026）增高。结论ADM能抑制切口中成纤维细胞的增生和胶原的过度分泌,减少瘢痕的形成。     

糖尿病大鼠创面愈合过程中成纤维细胞增殖及Ⅰ型胶原合成减少

目的：观察链脲佐菌素诱导的大鼠糖尿病复合创伤修复过程中成纤维细胞增殖与胶原合成的变化。 方法： 实验采用Wistar大鼠112只，随机分对照组和模型组。模型组大鼠腹腔注射链脲佐菌素（STZ）55 mg/kg，3周后各组动物复合背部2.04 cm²全厚皮切除形成伤口。观察创面愈合时间和愈合率；采用HE染色和免疫组化法观察成纤维细胞和增殖细胞核抗原(PCNA)表达水平；采用苦味酸-天狼星红染色和图像分析技术观察创面Ⅰ、Ⅲ型胶原含量及Ⅰ/Ⅲ型比值。 结果： STZ诱发的糖尿病大鼠复合创伤后创面的愈合时间为(27.13±1.81)d，明显长于对照组（15.25±1.67）d，P<0.01；模型组在创伤第3、7和15 d创面愈合率明显低于对照组，分别P<0.01；在3、5、7和9 d模型组创面成纤维细胞数量和PCNA表达也明显少于对照组，分别P<0.05和P<0.01。两组创面在不同时点上Ⅰ型胶原分布均呈递增趋势，但对照组明显多于模型组，分别P＜0.05；尽管对照组在创伤3、7 d Ⅲ型胶原含量高于模型组，但在创伤3、7和11 d模型组Ⅰ/Ⅲ胶原比值都明显低于对照组，分别P＜0.01。 结论： STZ可能通过影响创面细胞增殖和创面胶原合成，从而导致创面愈合迟缓。     

Therapeutic Effects of Topical Application of Ozone on Acute Cutaneous Wound Healing

This study was undertaken to evaluate the therapeutic effects of topical ozonated olive oil on acute cutaneous wound healing in a guinea pig model and also to elucidate its therapeutic mechanism. After creating full-thickness skin wounds on the backs of guinea pigs by using a 6 mm punch biopsy, we examined the wound healing effect of topically applied ozonated olive oil (ozone group), as compared to the pure olive oil (oil group) and non-treatment (control group). The ozone group of guinea pig had a significantly smaller wound size and a residual wound area than the oil group, on days 5 (P<0.05) and 7 (P<0.01 and P<0.05) after wound surgery, respectively. Both hematoxylin-eosin staining and Masson-trichrome staining revealed an increased intensity of collagen fibers and a greater number of fibroblasts in the ozone group than that in the oil group on day 7. Immunohistochemical staining demonstrated upregulation of platelet derived growth factor (PDGF), transforming growth factor-β (TGF-β) and vascular endothelial growth factor (VEGF) expressions, but not fibroblast growth factor expression in the ozone group on day 7, as compared with the oil group. In conclusion, these results demonstrate that topical application of ozonated olive oil can accelerate acute cutaneous wound repair in a guinea pig in association with the increased expression of PDGF, TGF-β, and VEGF.     

基于ILK信号通路探究马桑水提取物影响大鼠烧伤创面愈合和瘢痕增生的机制

目的:基于整合素连接激酶(ILK)调控的转化生长因子β1(TGF-β1)和PI3K/AKT调控的ILK等互通信号通路,探究马桑水提取物(CSME)促进大鼠烧伤创面早期愈合和抑制后期瘢痕过度增生的机制。方法:随机将180只SD雌性大鼠(180~200 g)分成正常对照(NC)组、凡士林(VL)组、磺胺嘧啶银(SS)组及CSME低、中和高剂量(CSME-L、CSME-M和CSME-H)组,每组各30只。大鼠以水合氯醛麻醉,腰背部II°烫伤造模后,每天分别在创面涂擦VL、SS和3种剂量的CSME烧伤软膏,计算创面愈合率(HR),于第7、14和21天随机选取各组动物10只,水合氯醛麻醉,切取创面皮肤,病理学观查,Western blot检测相关蛋白表达,RT-qPCR检测相关mRNA表达,培养成纤维细胞检测其胶原收缩率(SK)。结果:在第7天,CSME各组创面中ILK、纤维连接蛋白(FN)、TGF-β1、α-平滑肌肌动蛋白(α-SMA)和整合素β1(integrin-β1,ITG-β1)的蛋白与mRNA表达呈现剂量依赖性强于VL和SS组,第21天,弱于VL和SS组(P<0.05);第14天,CSME各组组织中I型胶原蛋白(Col I)的蛋白与mRNA表达呈剂量依赖性强于VL和SS组,第21天则弱于VL和SS组(P<0.05),而III型胶原蛋白(Col III)的蛋白与mRNA表达呈剂量依赖性弱于VL和SS组,第21天则强于VL和SS组(P<0.05);随时间推移,VL和SS组成纤维细胞的SK持续升高,96 h达高峰,而CSME各组在24 h和48 h呈剂量依赖性高于VL和SS组,在96 h呈剂量依赖性低于VL和SS组(P<0.05)。结论:CSME早期促进烧伤创面愈合,后期抑制创面瘢痕过度增生,其机制与其多成分多靶点影响ILK调控的TGF-β1和PI3K/AKT调控的ILK信号通路及ColⅠ与Col III表达比的调控相关。     

The role of integrin-β/FAK in cyclic mechanical stimulation in MG-63 cells

Objective: This study aims to explore the function of Integrin-β/FAK in the mechanical signal transduction and the connection with downstream ERK signal pathways. Methods: Human osteosarcoma MG63 cell lines were used in this study. The effects of mechanical strain on the Integrin-β₁ expression, FAK and ERK signal pathway in Human osteosarcoma MG63 cells were detected using RT-PCR and Western-blotting methods. The localization of FAK in Human osteosarcoma MG63 cells were determined using immunofluorescent method. The interaction between Integrin-β₁ and FAK were detected by using co-immunoprecipitation method. Results: The expression of Integrin-β₁ shows a notable bimodel distribution, mechanical strain stimulation can promote Integrin-β₁ expression and the phosphorylation of FAK and ERK, mechanical strain activated FAK and ERK mediated by Integrin-β₁. Conclusion: Integrin-β₁ may play an important role in osteoblast proliferation differentiation process, it might feel external strain stimulation through ECM composition and makes FAK phosphated through the interaction with FAK, thus causing a series of activation of signal molecules. Finally it reduces MAPK (ERK) activation and cellular responses to finish mechanical signal transduction.     

纳米银－猪脱细胞真皮基质敷料治疗浅Ⅱ度烧伤创面的临床观察

目的观察纳米银－猪脱细胞真皮基质敷料在临床上治疗浅Ⅱ度烧伤创面的临床疗效。 方法2014年1月到2015年12月,选取北京军区总医院烧伤整形科收治的浅Ⅱ度烧伤患者90例,按入院顺序依次编码,采用随机排列数字表法将90例患者分为纳米银敷料组、猪脱细胞真皮基质敷料组及纳米银－猪脱细胞真皮基质敷料组,每组30例。患者入院当天,拍照计算创面面积,并用咽拭子取创面分泌物作细菌培养,行创面清创术,分别在创面上敷以纳米银敷料、猪脱细胞真皮基质敷料以及纳米银－猪脱细胞真皮基质敷料。于治疗后第5天,用咽拭子取创面分泌物作细菌培养;采用痛觉评分标准,通过询问与观察患者换药时的痛觉情况,评估患者换药时痛觉评分。于治疗后第7天,拍照计算创面面积,计算创面愈合率。记录创面最终愈合时间。数据比较采用单因素方差分析、χ²检验及SNK－q检验。 结果纳米银敷料组、猪脱细胞真皮基质敷料组及纳米银－猪脱细胞真皮基质敷料组在治疗后第5天创面细菌培养阳性数结果分别为2例(6.6%)、9例(30.0%)、1例(3.3%),3组结果比较,差异有统计学意义(χ²＝10.962,P＝0.004);纳米银－猪脱细胞真皮基质敷料组的细菌培养阳性数结果显著优于猪脱细胞真皮基质敷料组,差异有统计学意义(χ²＝7.680,P＝0.006)。纳米银敷料组、猪脱细胞真皮基质敷料组及纳米银－猪脱细胞真皮基质敷料组治疗后第5天的痛觉评分[(8.6±0.5)、(6.6±0.8)、(0.6±1.3)分],治疗后第7天的创面愈合率[(61.67±18.22)%、(86.77±15.32)%、(99.80±0.56)%],创面愈合时间[(11.5±1.3)、(10.3±0.7)、(7.3±0.7)d],组间差异均有统计学意义差异(F＝201.7、19.9、55.7,P值均小于0.05);纳米银－猪脱细胞真皮基质敷料组治疗后第5天的痛觉评分显著低于其余两组,差异均有统计学意义(P值均小于0.05);治疗后第7天的创面愈合率明显优于其余两组,差异均有统计学意义(P值均小于0.05);创面愈合时间显著短于其余两组,比较差异均有统计学意义(P值均小于0.05)。 结论纳米银－猪脱细胞真皮基质敷料具有抗感染、促进创面愈合及减轻换药痛觉的作用。     

肝素化羊脱细胞真皮基质对深Ⅱ度烧伤创面组织中血管内皮生长因子和层粘连蛋白的影响

目的研究肝素化羊脱细胞真皮基质(ADM)促进深Ⅱ度烧伤创面愈合的作用机制。 方法制作大鼠深Ⅱ度烧伤模型,随机分为碘伏纱布组、猪ADM组、羊ADM组和肝素化羊ADM组,每组40只。在伤后不同时间段,观察各组创面愈合率,同时在各个时间段处死大鼠8只,取创面组织标本做RT-qPCR和蛋白质印迹法检测各组创面组织中血管内皮生长因子(VEGF)和层粘连蛋白(LN)的mRNA与蛋白表达情况。对数据行单因素方差分析和t检验。 结果肝素化羊ADM组深Ⅱ度创面的愈合率优于碘伏纱布组。RT-qPCR检测发现,VEGF的mRNA在各组伤后3 d的创面组织中表达较对照组高,各组间的比较发现,伤后7 d,肝素化羊ADM组的VEGF的mRNA表达量明显高于碘伏纱布组(t＝80.83,P<0.05),羊ADM组和猪ADM组在伤后7 d也较碘伏纱布组的表达量高(t＝60.80、53.42,P值均小于0.05);蛋白质印迹法检测肝素化羊ADM组伤后3 d时VEGF蛋白表达量较多,随后便逐渐减少,伤后21 d时蛋白表达较少。伤后3 d,碘伏纱布组LN在创面的表达明显下降,在之后的治疗过程中,LN的表达逐渐恢复,伤后14 d的表达量明显高于伤后3 d(t＝19.5,P<0.001),伤后21、28 d的表达量与伤后14 d相差不明显。蛋白质印迹法检测发现肝素化羊ADM组LN在伤后3 d便开始出现表达量逐渐增多,伤后21 d时出现蛋白表达较多。 结论肝素化羊ADM能够保护创面及促进创面组织中VEGF和LN的表达,进而促进创面愈合。     

Hyperbaric Oxygen Therapy Attenuates Burn-Induced Denervated Muscle Atrophy

Background: Neuronal apoptosis and inflammation in the ventral horn of the spinal cord contribute to denervated muscle atrophy post-burn. Hyperbaric oxygen therapy (HBOT) exerts anti-inflammation and neuroprotection. Furthermore, hypoxia-inducible factor (HIF)-1α has been reported to promote inflammation and apoptosis. We investigated the therapeutic potential of HBOT and the role of HIF-1α post-burn.Methods: Sprague-Dawley rats were divided into three groups: a control group, an untreated burn group receiving burn and sham treatment, and a HBOT group receiving burn injury and HBOT. The burn injury was induced with 75ºC ± 5ºC at the right hindpaw. HBOT (100% oxygen at 2.5 atmosphere, 90 min/day) and sham HBOT (21% oxygen at 1 atmosphere, 90 min/day) was started on day 28 after burn injury and continued for 14 treatments (days 28-41). Incapacitance (hind limb weight bearing) testing was conducted before burn and weekly after burn. At day 42 post-burn, the gastrocnemius muscle and the spinal cord ventral horn were analyzed.Results: HBOT improved burn-induced weight bearing imbalance. At day 42 post-burn, less gastrocnemius muscle atrophy and fibrosis were noted in the HBOT group than in the untreated burn group. In the ventral horn, HBOT attenuated the neuronal apoptosis and glial activation post-burn. The increases in phosphorylated AKT/mTOR post-burn were reduced after HBOT. HBOT also inhibited HIF-1α signaling, as determined by immunofluorescence and western blot.Conclusions: HBOT reduces burn-induced neuronal apoptosis in the ventral horn, possibly through HIF-1α signaling.     

Effect of SACCHACHITIN on keratinocyte proliferation and the expressions of type I collagen and tissue-transglutaminase during skin wound healing

SACCHACHITIN is a skin wound-healing membrane made of residual fruiting body of Ganoderma tsugae. Its effect on proliferating cell nuclear antigen (PCNA) expression in actively proliferating cells, type I collagen expression and tissue remodeling in the healing tissue, and the association of tissue-transglutaminase (t-TGase) with wound healing were investigated by immunohistochemical staining. The results demonstrated that PCNA expressed in keratinocytes since day 1 in the SACCHACHITIN group and persisted during entire healing process. In contrast, it was barely detectable on day 3 in the control group. At keratinocyte layer, the SACCHACHITIN group exhibited more type I collagen than did the control group since day 1. At scar tissue, type I collagen was positively stained in the SACCHACHITIN group since day 7 but not in the control group till day 12. Furthermore, t-TGase was strongly expressed on the inner wall of angiogenic vessels on day 5 of the control group but not on that of the SACCHACHITIN group until day 10. The earlier expressions of PCNA and type I collagen in the keratinocyte layer may lead to accelerated skin wound healing. In addition, the later expression of t-TGase, an indicator of apoptosis, on the inner wall of angiogenic capillaries in the SACCHACHITIN group may indicate a longer period of blood supply to the wound area, thus facilitating wound healing. These observed phenomena might underline the beneficial effects of SACCHACHITIN membrane on rapid wound healing.     

碱性调宁蛋白对糖尿病大鼠烧伤创面成纤维细胞增殖与凋亡的影响

目的探讨碱性调宁蛋白在糖尿病大鼠烧伤创面愈合过程中的表达情况及其对成纤维细胞增殖与凋亡的影响。 方法选64只雄性Wistar大鼠,按随机数字表法分为实验组和对照组,每组32只。实验组大鼠,腹腔注射链脲佐菌素60 mg/kg,制作糖尿病大鼠模型;对照组大鼠,腹腔注射0.9%氯化钠溶液1.0 mL。适应性饲养2周后将2组大鼠均制作为深Ⅱ度烫伤模型(以下称为烧伤),伤后立即液体复苏、常规治疗。观察两组大鼠的创面愈合时间及在伤后第7、14、21、28天的创面愈合率,创周切取组织、制作切片,行苏木精-伊红染色观察组织形态学变化,免疫组织化学SP法分析碱性调宁蛋白表达,成纤维细胞增殖细胞核抗原(PCNA)的表达,原位末端转移酶标记(TUNEL)法测定成纤维细胞凋亡情况。对数据行方差分析和t检验。 结果实验组大鼠创面平均愈合时间[(25.6±2.8) d]长于对照组大鼠创面平均愈合时间[(19.1±2.1) d],差异有统计学意义(t＝10.49,P<0.05);在伤后第7、14、21、28天,实验组大鼠创面愈合率分别为(18.8±4.6)%、(37.6±7.9)%、(66.3±11.4)%、(85.4±4.8)%,明显小于对照组[(40.8±4.5)%、(62.1±6.7)%、(90.2±9.7)%、(95.4±5.5)%,差异均有统计学意义(t＝9.69、6.69、4.51、3.87,P值均小于0.05)。伤后第7、14、21天,两组大鼠创面逐渐愈合,创面组织中成纤维细胞、新生微血管均逐渐增多,且对照组大鼠未愈合创面组织中成纤维细胞、新生微血管均较实验组增多。伤后第7、14、21、28天,实验组大鼠碱性调宁蛋白在烧伤创面组织中的表达分别为1.43±0.07、1.17±0.11、0.73±0.06、0.77±0.07,均高于对照组大鼠(0.75±0.09、0.60±0.10、0.63±0.06、0.64±0.10),差异均有统计学意义(t＝17.58、10.92、3.44、3.10,P值均小于0.05)。实验组大鼠成纤维细胞PCNA在烧伤创面组织中表达分别为654.19±102.80、820.84±112.86、766.68±156.63、232.75±55.37,较对照组(766.85±73.30、1322.22±121.44、1112.35± 142.32、740.79±106.90)低,差异均有统计学意义(t＝2.52、8.55、4.62、11.94,P值均小于0.05)。伤后第7、14、21、28天,实验组细胞凋亡数分别为58.51±10.89、41.53±9.95、27.28±6.58、20.13±4.23,较对照组(30.70±6.41、25.31±5.74、17.46±5.36、15.29±4.10)高,差异均有统计学意义(t＝6.23、4.00、3.27、2.32,P值均小于0.05)。 结论碱性调宁蛋白在糖尿病大鼠烧伤创面愈合过程中表达水平增高,可能调控创面组织中成纤维细胞的增殖和(或)凋亡水平,是糖尿病创面延迟愈合的原因之一。     

Human uterine leiomyoma cells: binding and growth responses to epidermal growth factor, platelet-derived growth factor, and insulin

The specific binding of epidermal growth factor (EGF), platelet-derived growth factor (PDGF) and insulin were measured in matching cultures of human leiomyoma and myometrial cells, along with the effects of these proteins on DNA and protein syntheses. Scatchard analyses of the binding data revealed that the EGF receptor sites/cell were significantly lower in leiomyoma than myometrial cultures. Two types of PDGF binding were observed when porcine PDGF was used, and one type was seen with human PDGF. By contrast to EGF, more PDGF receptor sites/cell were found in leiomyoma than myometrium but the receptor affinity was higher in the latter. Insulin binding was similar among the myometrial and leiomyoma cells. Protein synthesis was stimulated 3-fold by EGF, PDGF, or insulin in both cell types. DNA synthesis, was higher in myometrial than leiomyoma cells in the basal state and was stimulated by EGF, insulin, or PDGF. A synergistic stimulation (p less than 0.02) of DNA synthesis was observed in both myometrial and leiomyoma cells when EGF was added with insulin. The addition of PDGF with insulin caused only additive stimulation of DNA synthesis. However, the addition of EGF with PDGF caused a synergistic decrease (p less than 0.05) in DNA synthesis by myometrial but no leiomyoma cells. Cultures of human vascular smooth muscle cells obtained from umbilical veins gave results similar to those from myometrium. These findings single out the EGF receptor and EGF, or perhaps an EGF-like growth factor, and to a lesser degree PDGF, as potential regulators of uterine leiomyomata.     

Immunosuppressive effects of mesenchymal stem cell transplantation in rat burn models

Objective: This study is to investigate the effects of mesenchymal stem cell (MSC) transplantation in burn treatment. Methods: Wharton’s Jelly was stripped from neonatal umbilical cord, and human umbilical cord MSCs were then cultured. Burn models were constructed in male SD rats weighted at 200 ± 5 g, and the rats were randomly divided into control and MSCs transplantation groups. The rats in transplantation group were injected subcutaneously with MSCs (2×10⁶) at 24 h after burning. Blood samples were collected at 0 d, 1 d, 2 d, 3 d, 5 d and 7 d after burning and the contents of white blood cells (WBC), C-reactive protein (CRP), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10 ) were detected. The wound healing rate at 7 d, 14 d, 21 d and 28 d together with the wound healing time were compared and analyzed statistically by analysis of variance (ANOVA). Results: WBC and CRP in control group increased significantly at 1 d and 2 d, 2 d and 3 d, respectively. IFN-γ, IL-6 and IL-10 levels in serum showed increasing till 5th day and TNF-α arrived its peak value at 7th day. By contrast, WBC, CRP, TNF-α, IL-6 and IL-10 in the MSCs transplantation group showed slight increase after burning and the differences were verified by statistically analysis. IFN-γ showed no significant difference between the two groups. MSCs transplantation group showed significantly higher wound healing rate at 14 d, 21 d, 28 d and showed shorter wound healing time than control. Conclusions: MSCs transplantation could suppress secondary inflammatory reaction by lowering inflammatory cytokines after burning, thus promoting wound healing and scald repair in burn animal model.     

A proline derivative-enriched methanol fraction from Sideroxylon obtusifolium leaves (MFSOL) stimulates human keratinocyte cells and exerts a healing effect in a burn wound model

It was previously demonstrated that the methanol fraction of Sideroxylon obtusifolium (MFSOL) promoted anti-inflammatory and healing activity in excisional wounds. Thus, the present work investigated the healing effects of MFSOL on human keratinocyte cells (HaCaT) and experimental burn model injuries. HaCaT cells were used to study MFSOL''s effect on cell migration and proliferation rates. Female Swiss mice were subjected to a second-degree superficial burn protocol and divided into four treatment groups: Vehicle, 1.0% silver sulfadiazine, and 0.5 or 1.0% MFSOL Cream (CrMFSOL). Samples were collected to quantify the inflammatory mediators, and histological analyses were performed after 3, 7, and 14 days. The results showed that MFSOL (50 μg/mL) stimulated HaCaT cells by increasing proliferation and migration rates. Moreover, 0.5% CrMFSOL attenuated myeloperoxidase (MPO) activity and also stimulated the release of interleukin (IL)-1β and IL-10 after 3 days of treatment. CrMFSOL (0.5%) also enhanced wound contraction, promoted improvement of tissue remodeling, and increased collagen production after 7 days and VEGF release after 14 days. Therefore, MFSOL stimulated human keratinocyte (HaCaT) cells and improved wound healing via modulation of inflammatory mediators of burn injuries.     

TGF-β3基因转染成纤维细胞促进大鼠深Ⅱ度烫伤创面愈合的实验研究

目的观察TGF-β3基因转染的成纤维细胞对深Ⅱ度大鼠烫伤模型创面愈合的影响。方法将90只Wistar成年大鼠随机分为模型对照组(C组)、表皮转化生长因子组(EGF组)和TGF-β3转基因成纤维细胞组(TGF-β3组),制作10% TBSA烫伤模型,各组给予对应治疗,检测各组大鼠的创面愈合率,HE染色观察烫伤创面组织变化,SP法观察烫伤皮肤组织中TGF-β3蛋白表达情况。结果烫伤后14d、21d,创面愈合率TGF-β3组最高,与EGF组及C组之间的差异有统计学意义(分别为P0.05,P0.01);烫伤后14d及21d,C组、EGF组与TGF-β3组之间TGF-β1及TGF-β2蛋白表达差异有统计学意义(分别为P0.05,P0.01);烫伤后1d到21d,TGF-β3组与C及EGF组之间的TGF-β3蛋白表达的差异均有统计学意义(P0.01)。结论 TGF-β3转基因转染成纤维细胞可能通过在创面上持续分泌TGF-β3蛋白,降低TGF-β1及TGF-β2的水平,从而促进创面的愈合。     

Wound-Healing and Potential Anti-Keloidal Properties of the Latex of Calotropis Procera (Aiton) Asclepiadaceae in Rabbits

Calotropis Procera (CP) has been used in the management of toothache, fresh skin burns, gum bleeding as well as others to make it qualify as a medicinal plant. This study was designed to assess its wound-healing property in rabbits and its potentials for anti keloidal activity.Fresh latex of Calotropis were obtained and evaluated phytochemically. Fifteen male rabbits were used and four excisional wounds were created on each rabbit. The rabbits were divided into five groups of three each. Group 1 was the negative control and received no treatment. The wounds of group 2 animals were treated with 2mL of Calotropis latex; group 3 with 2mL honey; and group 4 with a mixture of 1ml honey and 1 mL of the latex. The animals in group 5 were given 2mg triamcinolone intramuscularly. All the groups had their wounds treated daily for 21 days. The wounds'' diameters were measured on the day of wound creation, thereafter on days 7, 14 and 21 post wound creation. Biopsies of the wounds were taken on days 3 and 21 and viewed histologically. Phytochemical study of the latex revealed the presence of glycosides, tannins and alkaloids. The wounds were found to be significantly (p<0.05) reduced in groups treated with 50% latex in honey and triamcinolone, respectively, on day 7 post wound creation while there was a significant (p<0.05) reduction in wound surface area in all treated groups on days 14 and 21 post wound creation. Histological findings in untreated group showed thick bundle of collagen fibres some of which had broad based configurations, reminiscent of keloid. The group treated with 2mL of Calotropis latex revealed the presence of florid granulation tissues on day 3 while there was a marked reduction in quantity and size of collagen fibres on day 21 post wound creation which was comparable with what was seen for the triamcinolone-treated group.The general effect of Calotropis latex on wound-healing was noted. Likewise it''s similarity to that of triamcinolone, an anti-keloidal agent; this makes it a probable candidate for future anti-keloidal study using a suitable model.     

Kupffer细胞在实验性肝癌形成中的作用

目的探讨Ｋｕｐｆｅｒ细胞对大鼠实验性肝癌形成的影响。方法应用免疫组化和原位分子杂交技术,对单纯用化学致癌剂二乙基亚硝胺（ＤＥＮＡ）及氯化钆（ｇａｄｏｌｉｎｉｕｍｃｈｌｏｒｉｄｅ,ＧＣ）填充Ｋｕｐｆｅｒ细胞后,同时给以ＤＥＮＡ所引起的大鼠肝癌形成过程中的增殖细胞核抗原（ＰＣＮＡ）、表皮生长因子（ＥＧＦ）、ｒａｓｐ２１、ｐ５３蛋白及ｃｍｙｃ基因表达进行了对比研究。结果结果显示：ＰＣＮＡ、ＥＧＦ表达在两组动物肝组织中未见明显差异;ＧＣ＋ＤＥＮＡ组ｒａｓｐ２１表达阳性率明显高于ＤＥＮＡ组;ｐ５３蛋白的表达在ＧＣ＋ＤＥＮＡ组略高于ＤＥＮＡ组,但出现时间则远较ＤＥＮＡ组为早,前者出现于第１１周,后者则在第１９周;ｃｍｙｃ基因表达强度ＧＣ＋ＤＥＮＡ组明显高于ＤＥＮＡ组。结论上述结果提示Ｋｕｐｆｅｒ细胞能够降低化学致癌剂对肝细胞的毒性损伤,从而在一定程度上对肝细胞的癌变过程可能具有抑制作用。     

Histopathological evaluation of the healing effects of human amniotic membrane transplantation in third-degree burn wound injuries

Skin burn injuries result in loss of its protective function as a barrier and leading to a high risk of infection. Therefore, effective treatments and healing of burn injuries are very important to prevent complications. Amniotic membrane (AM) as a biological dressing inhibits the loss of vital fluids, water, and protein. The aim of this study was to compare the healing effects of AM and silver sulfadiazine (SSD) ointment in third-degree burn injuries in experimental rat model. Fifty-four male Sprague–Dawley rats were divided randomly into three equal groups. After induction of third-degree burn, transplantation of human AM (HAM) and SSD ointment used on wound area for treatment groups. The third group was considered as control. At days 7, 14, and 21, histopathological evaluation of burn wound area was performed using light microscopy. After 21 days, burn injury in HAM group showing lack of enough wound contraction and decrease in wound area in comparison to SSD group. Also, the healing effects were demonstrated using decline of inflammatory cell infiltration and enhanced epithelium after 21 days. The total wound score was significantly higher in the HAM group than the control and SSD ointment groups, during and at the end of the experiment (P?<?0.05). On day 21, significantly lesser inflammatory cell infiltration was noticed in the control group (P?<?0.05). Our findings showed that HAM can be used successfully as a biological treatment for experimental third-degree burn injury in animal model.     

体外培养表皮干细胞复合高分子支架原位修复深度烧伤创面的研究

目的探讨表皮干细胞联合成纤维细胞-丝素蛋白纳米纤维活性支架体内培养,对Ⅲ度烧伤创面的修复和再生作用。方法（1）表皮干细胞的培养和表征：采用快速贴壁法分离和培养表皮干细胞（Epidermal Stem Cell,ESC）。将表皮干细胞分别在经Ⅳ型胶原蛋白修饰的和未经修饰的培养瓶中,或通过悬浮法培养,研究表皮干细胞的生长特性;以β1整合素和细胞角蛋白CK19免疫荧光染色实验考察细胞表型。（2）活性支架的体外构建和对大鼠Ⅲ度创面的修复作用研究：体外构建成纤维细胞-丝素蛋白纳米纤维活性支架;采取同体对照法,在20只Sprague—Dawley大鼠（sD大鼠）背部制作两个Ⅲ度切痂创面。左侧创面采用体外培养的自体表皮干细胞联合成纤维细胞一丝素蛋白纳米纤维活性支架移植入创面,作为组织工程移植物组;右侧创面采用凡士林纱布敷料覆盖,作为凡士林纱布敷料组。考察组织工程移植物对大鼠Ⅲ度创面的愈合作用。结果以快速贴壁法能够有效地分离得到表皮干细胞,细胞在经Ⅳ型胶原蛋白修饰的培养瓶中生长10d后融合,数目达到5．1×10^5／cm^2。免疫荧光实验表明细胞表面抗原呈β1整合素和角蛋白免疫CK19成阳性,证明分离得到的细胞为表皮干细胞。成纤维细胞能够在丝素蛋白纳米纤维中扩增并分泌细胞外基质,14d后与丝素蛋白纳米纤维形成活性支架。对大鼠Ⅲ度烧伤创面的修复实验表明,组织工程移植物组的创面在第14天和第22天的平均愈合效率为66％和93％,高于凡士林纱布敷料组（32％和69％）,P〈0．05。组织工程移植物组的创面平均愈合天数为21d,低于凡士林纱布敷料组（31d）,P〈0．05。结论通过Ⅳ型胶原蛋白黏附法,能够分离得到表皮干细胞,并且其在Ⅳ型胶原蛋白表面修饰的培养瓶中的生长活力较高。大鼠Ⅲ度创面的修复实验表明,组织工程移植物,即表皮干细胞联合成纤维细胞-丝素蛋白纳米纤维支架,能够修复Ⅲ度创面,再生皮肤表真皮结构完整;并且与凡士林纱布敷料相比,能够提高创面的愈合效率,减少创面的愈合时间。