1064nm皮秒激光联合表皮干细胞外泌体治疗黄褐斑临床疗效观察

分析黄褐斑应用表皮干细胞外泌体结合1064nm皮秒激光治疗效果。方法 2022-3~2023-3黄褐斑患者102例,比对组51例1064nm皮秒激光。分析组51例(1064nm皮秒激光联合表皮干细胞外泌体);比较不同方案治疗效果差异。结果 分析组在治疗效果高于比对组,P<0.05。两组在安全性方面互比,无差异性,P>0.05。分析组在皮损消失时间、皮损颜色淡化时间均短于比对组,P<0.05。分析组在治疗满意度高于比对组,P<0.05。结论 在1064nm皮秒激光治疗基础上增加表皮干细胞外泌体可进一步提高对黄褐斑疾病可达到综合治疗目的,而且疗效确切,易获得患者认可,安全可行性高,值得在临床中推广与运用。

新型强脉冲光联合表皮干细胞外泌体治疗黄褐斑临床疗效观察

目的:探讨新型强脉冲光(Advanced optimal pulsed technology,AOPT)联合表皮干细胞外泌体(Extracellular vesicles released from epithelial stem cells,EVS)治疗黄褐斑的临床疗效及安全性。方法:选2019年3月-2021年12月在笔者医院接受治疗的89例黄褐斑患者,随机分为实验组(AOPT+EVS治疗组)和对照组(AOPT治疗组),疗程24周,比较治疗前后黄褐斑面积和严重指数(Melasma area severity index,MASI)评分,医生整体评价(Physician’s global assessment,PGA)、患者满意度及出现的不良反应。结果:实验组治疗后MASI评分由(18.1±8.0)分下降至(4.2±3.6)分;对照组治疗后MASI评分由(17.6±6.4)分下降至(5.8±3.8)分,两组治疗前后差异有统计学意义(P<0.05),两组间治疗前后差值比较差异有统计学意义(P<0.05)。两组PGA评分均有改善,实验组改善率97.7%,对照组改善率93.3%;患者满意度实验组88.6%,高于对照组的80.0%。治疗期间不良反应轻微,未观察到严重不良反应。结论:新型强脉冲光治疗黄褐斑疗效确切,术后配合EVS外用可以在一定程度上提高治疗效果。

2，3，7，8-四氯二苯并二噁英对体外表皮细胞增殖的影响

目的研究2，3，7，8-四氯二苯并二噁英（TCDD）对人角质形成细胞（HACAT）增殖的影响。方法选用5个不同染毒浓度（7×10^-12～7×10^-8 mol／L），对生长至60％～70％汇合的HACAT细胞进行体外染毒，并在培养的不同时间点（6、12、24、48h）进行检测，分别通过形态学、克隆形成实验和噻唑蓝（MTT）比色法观察TCDD对细胞增殖的影响。结果TCDD对细胞形态的影响并不明显；对克隆形成率有明显的促进作用，但剂量-效应关系不明显；MTT检测结果显示，TCDD对HACAT细胞的生长增殖有促进作用，且与时间、浓度存在一定的依赖关系，染毒6、12、24h，细胞增殖率随浓度的增加而增加，相关系数分别为0．561，0．687，0．720，差异有统计学意义（P〈0．01）；但48h组，细胞的增殖率不再增加，相反对细胞的生长表现为抑制作用。结论TCDD对细胞形态无明显影响，但在一定染毒时间范围内（6～24h），对体外HACAT细胞的生长增殖有促进作用，且与时间、浓度存在正相关。

Ex vivo differentiation of multipotent adult progenitor cells to skin epidermal cells

Objective： By establishing the indirect contact co-culture system, we studied the in vitro condition for MAPCs differentiating into epidermal cells and the transformation of MAPCs into epidermal cell phenotype. Methods： Cell culture insert membrane was used for substitute basal membrane and MAPCs, fibroblast cells （FCs） and mixture of MAPCs and epidermal cells and FCs were separately implanted into 2 sides of it. PKH26 was used to label cloned MAPCs; type IV collagen rapid adhering method was used to isolate and culture the skin epidermal cells from l-day-old SD rat. Results： Part of the MAPCs transformed into cells expressing keratin in the presence of peripheral epithelia and FCs. Type Ⅳ collagen rapid adhering method successfully selected rats＇ epidermal stem cells. The mixture of the 2 kinds of cells or indirect culture might promote the differentiation through mesenchymal factors secreted by dermis FC. Conclusion： We were the first to have established the in vitro model of MAPCs differentiation into epidermal cells, in which MAPCs were transformed into epithelium-like cells.

Exogeneous energy supply and excitability of cells in embryonic atypical epidermis of Cynops cultured in vitro

Cells of in vitro cultured epidermis explants of ectoderm isolated at early gastrula stage,showed only weak excitability or even non-excitable at 6V when examined electrophysiologically.If non-excitable explants were treated with 100 mM glucose,the action potential (AP) appeared and within 1 hr reached its maximum.At the same time,their stimulus threshold became lowered gradually.And,if the glucose was washed out,AP gradually disappeared.If explants were treated with glucose of different concentrations,the percentage of explants which displayed AP increased with the increase of glucose concentration.When explants with approximately the same original stimulus threshold were treated with glucose of different concentrations,the stimulus threshold became lowered more in the more concentrated solution.If explants with different original stimulus thresholds were treated with glucose of the same concentration,the lowering of stimulus threshold was more obvious in those with higher original stimulus threshold.Other energy supplying substances used showed similar effect.

Anti-angiogenesis effect of generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide on breast cancer in vitro

Objective:To study the effects of the generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide(G4PAMAM/VEGFASODN) compound on the expressions of vascular endothelial growth factor(VEGF) and its mRNA of breast cancer cells and on the inhibition of vascular endothelial cells. Methods:We examined the morphology of G4PAMAM/VEGFASODN compound and its pH stability,in vitro transfection efficiency and toxicity,and the expressions of VEGF and its mRNA. Methyl thiazolyl tetrazolium assay was used to detect the inhibitory function of the compound on vascular endothelial cells. Results:The compound was about 10 nm in diameter and was homogeneously netlike. From pH 5 to 10,it showed quite a buffered ability. The 48-h transfection rate in the charge ratio of 1:40 was 98.76%,significantly higher than that of the liposome group(P<0.05). None of the transfection products showed obvious toxicity on the cells. The expressions of both VEGF protein and its mRNA after G4PAMAM/VEGFASODN transfection decreased markedly. Conclusion:With a low toxicity,high safety,and high transfection rate,G4PAMAM/VEGFASODN could be a promising gene vector. Specifically,it inhibits VEGF gene expression efficiently,laying a basis for further in vivo animal studies.

Effects of moxibustion pretreatment on extracellular signal-regulated kinase signaling transduction pathway in the gastric tissues of rats with gastric mucosal damage

Objective： To observe the effects of moxibustion pretreatment on the protein expressions of epidermal growth factor receptor （EGFR）, phosphorylation extracellular signal-regulated kinase I/2 （p-ERKI/2） and activated protein-1 （AP-2）, the key factors of extracellular signal-regulated kinase signaling transduction pathway in gastric tissue of rats with stress-induced gastric mucosal damage, and to discuss the mechanisms of moxibustion therapy in promoting the restoration of damaged gastric mucosa. Methods： Thirty Sprague-Dawley （SD） rats were randomly divided into a normal group, a model group, and a moxibustion group using the random digits table, 10 in each group. Except the rats in the normal group, rats in the other two groups were used to make stress-induced gastric mucosal damage model using restraint and cold stress. Before modeling, rats in the moxibustion group were alternately treated with moxibustion a/t Zusanli （ST 36） and Zhongwan （CV 12）, or Pishu （BL 20） and Weishu （BL 22）, once a day, for a total of 8 d. Histolo^cal changes of gastric mucosa were observed under the light microscopy, the expression of gastric tissue p-ERKI/2 was detected by immunohistochemistry assay, and the protein levels of EGFR and AP-I were measured by Western blots. Results： Compared with rats in the normal group, gastric mucosal damage was more serious, and protein expressions of gastric tissue EGFR, p-ERK1/2 and AP-1 increased in the model group （P〈0.01, P〈O.05, P〈0.05）. Compared with rats in the model group, gastric mucosal damage was milder, and protein expressions of gastric tissue EGFR, p-ERK1/2 and AP-1 increased in the moxibustion group （all P〈0.01）. Conclusion： Moxibustion at Zusanli （ST 36）, Zhongwan （CV 12）, Pishu （BL 20） and Weishu （BL 21） could increase EGFR, p-ERK1/2 and AP-1 expression levels in gastric tissue of stress-induced gastric mucosal damage rats, maintain the information transfer function of ERK signaling transduction pathway, and promote restoration of gastric mucosal damage.