Exploring the potential mechanism of WuFuYin against hypertrophic scar using network pharmacology and molecular docking

BACKGROUND Hypertrophic scar(HTS)is dermal fibroproliferative disorder,which may cause physiological and psychological problems.Currently,the potential mechanism of WuFuYin(WFY)in the treatment of HTS remained to be elucidated.AIM To explore the potential mechanism of WFY in treating HTS.METHODS Active components and corresponding targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.HTSrelated genes were obtained from the GeneCards,DisGeNET,and National Center for Biotechnology Information.The function of targets was analyzed by performing Gene Ontology and Kyoto Encyclopaedia of Genes and Genome(KEGG)enrichment analysis.A protein+IBM-protein interaction(PPI)network was developed using STRING database and Cytoscape.To confirm the high affinity between compounds and targets,molecular docking was performed.RESULTS A total of 65 core genes,which were both related to compounds and HTS,were selected from multiple databases.PPI analysis showed that CKD2,ABCC1,MMP2,MMP9,glycogen synthase kinase 3 beta(GSK3B),PRARG,MMP3,and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit gamma(PIK3CG)were the hub targets and MOL004941,MOL004935,MOL004866,MOL004993,and MOL004989 were the key compounds of WFY against HTS.The results of KEGG enrichment analysis demonstrated that the function of most genes were enriched in the PI3K-Akt pathway.Moreover,by performing molecular docking,we confirmed that GSK3B and 8-prenylated eriodictyol shared the highest affinity.CONCLUSION The current findings showed that the GSK3B and cyclin dependent kinase 2 were the potential targets and MOL004941,MOL004989,and MOL004993 were the main compounds of WFY in HTS treatment.

A Case Report of Concurrent Acne-Related Occurrence Complications: Telangiectasia, Post-Inflammatory Erythema, Post-Inflammatory Hyperpigmentation, and Atrophic and Hypertrophic Scars

Prior to his initial diagnosis, a 21-year-old male had been experiencing facial acne for two years and had been treated by a doctor in private practice. The patient visited our department because the clinical manifestations of mandibular acne did not improve. At the time of initial examination, telangiectasia (TE), post-inflammatory erythema (PIE), post-inflammatory hyperpigmentation (PIH), atrophic scars (ASs), and a hypertrophic scar (HS) with induration were observed on the right neck. We diagnosed this as an acne vulgaris complication. HS lesions were topically treated by injecting triamcinolone acetonide, and the patient was prescribed 8.1 g/day of oral Saireito (Japanese herb). Adapalene benzoyl peroxide gel and topical tacrolimus hydrate ointment were used to treat PIE and TE. Both HSs and PIE improved;however, TE and AS did not improve. Currently, the patient is under observation. We consider this to be a very rare concurrent occurrence of diverse complications of acne vulgaris, and present the following case study.

Dynamic changes of autophagy during hypertrophic scar formation and the role of autophagy intervention

Background:The role of autophagy in the formation of hypertrophic scars(HS)remains unclear.This study aimed to explore the role and potential mechanism of autophagy during the development of HS.Methods:RNA and protein expression levels of Beclin-1,p62,and LC3II in normal skin tissues and HS specimens from different patients were examined.Autophagy inducers and inhibitors were used to cure established HS in rabbit ears,and the expression of Beclin-1,p62,and LC3II at the RNA and protein level was determined.Lastly,the effects of autophagy inducers and inhibitors on HS development were analyzed.Results:Compared to normal skin tissues,the expression of LC3II and Beclin-1 was higher(P<0.05),while that of p62 was lower(P<0.05)in HS tissues.In addition,the LC3II/LC3I ratio was increased during HS formation,and the altered expression of the three proteins stabilized after one year.Administration of autophagy inducers enhanced the formation of HS as well as the expression levels of LC3II and Beclin-1 but decreased p62 expression.Meanwhile,administration of autophagy inhibitors increased the expression of LC3II,Beclin-1,and p62,along with reduced HS formation.Conclusion:Autophagic activity increased during HS initiation and subsequent stabilization.In addition,autophagy inhibitors were able to inhibit HS formation by suppressing autophagy,whereas autophagy inducers promoted scar hyperplasia by enhancing autophagy。

Inflammation and cutaneous nervous system involvement in hypertrophic scarring

This study aimed to use a mouse model of hypertrophic scarring by mechanical loading on the dorsum of mice to determine whether the nervous system of the skin and inflammation participates in hypertrophic scarring. Results of hematoxylin-eosin and immunohistochemical staining demonstrated that inflammation contributed to the formation of a hypertrophic scar and increased the nerve density in scar tissue.Western blot assay verified that interleukin-13 expression was increased in scar tissue. These findings suggest that inflammation and the cutaneous nervous system play a role in hypertrophic scar formation.

NON-LINEAR SPECTRAL IMAGING MICROSCOPY STUDIES OF HUMAN HYPERTROPHIC SCAR

Skin scar is unique to humans,the major significant negative outcome sustained after thermal injuries,traumatic injuries,and surgical procedures.Hypertrophic scar in human skin is investigated using non-linear spectral imaging microscopy.The high contrast images and spectroscopic intensities of collagen and elastic fibers extracted from the spectral imaging of normal skin tissue,and the normal skin near and far away from the hypertrophic scar tissues in a 10-year-old patient case are obtained.The results show that there are apparent differences in the morphological structure and spectral characteristics of collagen and elastic fibers when comparing the normal skin with the hypertrophic scar tissue.These differences can be good indicators to differentiate the normal skin and hypertrophic scar tissue and demonstrate that non-linear spectral imaging microscopy has potential to noninvasively investigate the pathophysiology of human hypertrophic scar.

The expression of Cyclin A and p21^(cip1)in fibroblast of hypertrophic scar

Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for intervention therapy of

Inhibition effects of a negative electret 5-FU patch on the growth of a hypertrophic scar

In this study,the hypertrophic scar（HS） model in rats was established.5-fluorouracil（5-FU）patch,-1000 V and-2000 V polypropylene（PP） electret 5-FU patches were prepared and applied onto the wound.The in vitro permeation experiment was performed using the Franz diffusion cell system to determine the permeation cumulative amount and retention amount of5-FU through/in scar skin.The inhibition effect of negative electret on growth of HS was studied by hematoxylin-eosin（HE） staining,Masson staining and the immunohistologicall methods.The permeation study indicated that a negative electret could enhance the permeation and retention of 5-FU through and in scar skin respectively.HE staining and Masson staining indicated a better effect for-1000 V and-2000 V electret 5-FU patches on HS inhibition after28 d post-wounding compared with 5-FU patch.The immunohistological study showed much more reduced expressions of collegan type I,collegan type III,TGF-β1 and HSP47 in scar tissue after application of negative electret 5-FU patches than those of 5-FU patch.A negative electret5-FU patch may be advantageous for HS treatment.

Novel Elastomeric Fabrics for the Treatment of Hypertrophic Burn Scars Using Polyhexamethylene Biguanide Antimicrobial Agents

In burn treatments,microorganisms on pressure garments during pressure therapy can prevent rehabilitation by causing functional,hygienic,and aesthetic difficulties. As bacterium is one of the most trouble-causing organisms,they can threaten patients causing infection during the long period of use of these garments.Novel burn pressure garments having durable antimicrobial property were developed using polyhexamethylene biguanide( PHMB)antimicrobial agent procedure on highly elastic nylon 66 /spandex fabrics in powernet,flat warp and weft knitted structures using paddry-cure method. Commercial wireless pressure sensors were used to control pressures at an acceptable medical range. Antimicrobial activity,wash durability,Fourier transform infrared spectroscopy(FTIR) and Scanning electron microscopy(SEM) analyses were conducted for the treated samples. Antimicrobial test results following AATCC 100 Test Method showed 99% reduction of bacteria for the fabric samples treated with PHMB. A small but significant decrease in antimicrobial activity was observed even after50 launderings. These treatments also yield good results to prevent odor,decrease infection by preventing and /or blocking microbial growth according to the antimicrobial mechanism and support reducing of scarring by providing a hygienic environment around the scar.

Expression of transforming growth factor-β_1 and its typeⅠ receptor in different phases of post-burn hypertrophic scars

Objective: To analyze and compare the expression pattern of the transforming growth factor-β1(TGF-β1) and its type I receptor (TGF-β RI ) in nounal human skin and various phases of post-burn hypertrophic scars (HTS). Method: The immunohistochemical ABC method was employed. Results: In nounal human skin, no evident immunoreactivity of TGF-β1 and TGF-β R I was observed. In activation phase of post-burn HTS, TGF-β R I and TGF-β1 were highly expressed in most dermal fibroblasts which seemed to be the same subset. However, in remission phase, no staining was seen in der mal fibroblasts. Conclusion: The formation of all may involve the increase of TGF-β responsiveness in fibroblasts The ac cumulation at the wound site and failure of apoptosis of over-resposive fibroblasts may contribute to the formation of HTS.

Expression of MMP-3 mRNA in hypertrophic scar fibroblasts and MMP-3 gene transfection

To investigate the molecular mechanism of extracellular matrix overdeposition in hypertrophic scar tissues and to explore MMPs gene therapy for hypertrophic scar. Methods: Hypertrophic scarderived and normal skin-derived fibroblasts were cultured and a recombinant retrovirus vector containing MMP-3 gene was constructed and then transfected into hypertrophic scar fibroblasts. Expressive level of MMP-3 mRNA was detected by dot blotting, and the activity of MMPs was determined by DNP-peptide.Results: Lower expression of MMP-3 mRNA and fewer DNP-peptide hydrolyzed fragments were observed in hypertrophic scar-derived fibroblasts compared with normal skin-derived fibroblasts. Transfection of MMP-3gene into hypertrophic scar-derived fibroblasts could enhance the expression of MMP-3 mRNA (3. 4 fold)and the de novo capacity to hydrolyze DNP-peptide (2. 1 fold). Conclusion: Overdeposition of extracellular matrix in hypertrophic scar tissue was related to low expression of MMP-3 due to its down-degradation of extracellular matrix. MMP-3 gene transfection could be a better way to treat hypertrophic scars by degrading extracellular matrix.

Effects of the He-Ne laser with various power densities on the growth of cultured fibroblasts in hypertrophic scar

Objective To explore the relationship between power density of He -Ne laser with the growth of fibroblasts in hypertroph ic scar (HS).Methods The cultured fibroblasts in HS were i rradiated with He -Ne laser(Wavelenth 632.8nm),various power densities such as50mW/cm 2 ,100mW/cm 2 and 150mW/cm 2 were respectively adopted once a day for 10minutes.After 1,3and 5times o f He -Ne laser for dose rate irradiation separatedly ,the cell count and cell circle analysis were counted and examined by trypa n blue staining and flow cytometry .Results The amount of cell after 1times of 50mW /cm 2 laser irradiation was markedly more than teh control(P<0.01).The amount of cells after 5times of 100mW/cm 2 ,3and 5times of 150mW/cm 2 laser irradiation was less than the controls(P <0.05).The result of cell circle analysis was corresponded with that of the cell count.Conclusion Both stimualtion and inhibition of He -Ne laser on the growth of scar fibroblasts can be obtained with vario us power densities.Power density of He -Ne laser associates with the effects on the growth of scar fibroblasts.

Hypertrophic Scar Formation and Wound Healing Modulation Fatty Acids as Modulators of Severe Scars

Scar tissue usually generates severe discomfort in the short and long term. Common symptoms include anesthetics sequelae, pruritus, joint malfunction, new wounds on the scar surface, and pain. There are several treatments for scars, like compression, topical or intralesional steroid infiltration, 5-fluorouracil, dermabrasion, and surgeries with new scar tissue. For adult patients, it is easier to choose the treatment. However, compression is commonly applied in children to prevent treatments that have adverse effects. This study reports the outcomes of 15 patients submitted to abdominoplasty, traumatic wounds and post-burn scar treatments, which showed significant changes after the continuous use of an ointment composed of petrolatum, cod liver oil, BHT, Chamomilla recutita (chamomile) oil, Helianthus annuus (sunflower) oil, and Prunus amygdalus dulcis (sweet almond) oil. As components of the stratum corneum, unsaturated fatty acids influence the cutaneous structural and immune status and permeability. They also interfere with the maturation and differentiation of the stratum corneum and inhibit the production of proinflammatory eicosanoids, reactive species (ROS and RNS), and cytokines, thereby influencing the inflammatory response and possibly wound healing. This article aims to share our experience with the regular use of an ointment in adult and pediatric patients for three months. The increase in proinflammatory cytokine production at wound sites, resulting in a noninvasive, therapeutical, and effective cutaneous wound healing and scarring modulation, may provide a physiopathological explanation for the fast improvement of scars.

Effect of ALA-PDT on the expressions of MMP-9, MMP-13 and TIMP-1 of hypertrophic scar model in rabbit ears

Objective: To investigate the effect of 5-aminolevulinic(ALA)-photodynamic therapy(PDT) on the expressions of MMP-9, MMP-13 and TIMP-1 of hypertrophic scar model in rabbit ears, and analyze the possible therapeutic mechanisms of ALA-PDT treatment to hypertrophic scars of rabbit ears. Methods: The experimental animals were randomly divided into normal control, negative control, high concentration of ALA-PDT, low concentration of ALA-PDT and PDT groups. The latter three groups received ALA-PDT treatment or PDT treatment once a week for 3 weeks. The specimens of the rabbits were collected respectively 1, 2 and 3 months after treatment to be used for RT-PCR and Western-blot test. Results: 1, 2 and 3 months after PDT treatment, the expressions of MMP-9 and MMP-13(including mRNA and protein) in hypertrophic scar tissues of three treatment groups were significantly higher than those of the negative control group(P<0.01), and the expression of TIMP-1 mRNA and protein of three treatment groups were significantly lower than that of the negative control group(P<0.01). There were also significant differences between high-concentration ALA-PDT treatment group and the low one(P<0.05). Conclusion: ALA-PDT is effective in treating hypertrophic scars of rabbit ears, and its possible therapeutic mechanisms are that ALA-PDT treatment generates oxidation activation effect to activate the activity of MMPs and induces the photoaging of fibroblasts of hypertrophic scar tissues of rabbit ears to inhibit the activity of TIMPs, which causes the up-regulation of MMPs and the down-regulation of TIMPs. Because of this, the degradation of collagen and ECM is accelerated and the formation of scars is suppressed.

Inhibitory effects of He-Ne laser repeated irradiation on collagen synthesis in hypertrophic scar-derived fibroblasts in culture

Objective To explore the inhibitory effects of He-Ne laser repeated irradiation on the collagen synthesis of cultured scar fibroblasts. Method Cultured fibroblasts derived from hypertrophic scars(HS) were irradiated with He-Ne laser for 30 minutes at various power densities(10,50,100 and 150 mW/cm2),once a day for 3 consecutive days.In 24 hours after repeated irradiation collagen production and type I procollagen mRNA level of fibroblasts were measured with the incorporation of 3H proline and blot hybridization techniques respectively.Results Collagen synthesis and type I procollagen mRNA level remained unchanged when the laser was irradiated at the power density of 10 mW/cm2 or 50 mW/cm2.Compared with control,collagen synthesis and type I procollagne mRNA level were significantly decreases at the power density of 100 mW/cm2 or 150mW/cm2(P< 0.05).Type I procollagen mRNA level at the power density of 150 mW/cm2 was lower than that at the 100 mW/cm2 (P< 0.05).Conclusion Repeated He-Ne laser irradiation at the power density of 100 mW/cm2 or 150 mW/cm2 can suppress collagen synthesis of cultured fibroblasts in HS.The cause of suppression may be associated with down regulation of type I procollagen mRNA expression.

STUDY ON FUNCTION OF FOCAL ADHENSIVE KINASE AND INTEGRIN α_1 IN HYPERTROPHIC SCAR FIBROBLASTS

Objective To study the function of focal adhesion kinase (FAK) in the formation of hypertrophic scar and its interrelationship with integrin α1. Methods Original fibroblasts from human hypertrophic scar and human normal dermis were cultured, and immunocytochemistry was applied to detect localization of expression of FAK and integrin α1 in hypertrophic scar and human normal skin fibroblasts. The expression of integrin α1 was detected before and after FAK antibody blocking hypertrophic scar fibroblasts (HSFB) 48 h later. Meanwhile the collagen synthesis was evaluated by [3H]-proline incorporation and HSFB cell proliferation was measured by MTT method. Results The expression of FAK and integrin α1 of hypertrophic scar fibroblasts was higher than that of the normal skin fibroblasts significantly (P<0.01). The expression of integrin α1 was reduced after FAK being blocked (P<0.01). Meanwhile the collagen synthesis of human scar-derived fibroblasts by [3H]-proline incorporation was depressed respectively (P<0.01). The cell proliferation was inhibited by using 1∶100 and 1∶200 FAK antibody with MTT method (P<0.01). Conclusion FAK is the key point of signal transmission pathway mediated by integrin α1, which regulates protein synthesis of integrin α1, it may play an important role in the proliferation and constriction of hypertrophic scar. FAK antibody can inhibit the collagen synthesis and cell proliferation of hypertrophic scar fibroblasts.

COMPARATIVE STUDY OF FIBRONECTIN GENE EXPRESSION IN TISSUES FROM HYPERTROPHIC SCARS AND DIABETIC FOOT ULCERS

Objective.To explore the expression characteristic of fibronectin gene in hypertr ophic scars and diabetic ul-cer tissues.Methods.The biopsies from normal skins,hypertrophic scars and diabetic foot ulc ers were taken.The tech-nique of quantitative polymerase ch ain reaction(PCR)was used to evaluate the gene express ion of fibronectin in the above biopsies.Results.Fibronectin gene expression was enhanced in hypertrophic scars and decreased in diabetic foot ul-cers compared with that in normal ski ns.Quantitative comparison showed about 2-fold increase of fibronectin mR-NA level in hypertrophic scars and ab out 3-fold decrease of fibronectin mRNA level in diabetic ulcers as com-pared with that in normal skins.Conclusions.Fibronectin gene expression is influenced by the tissue environment.Di fferent expression and synthesis of fibronectin may cause d ifferent outcomes in wound healing.

基于sCARS的淮北平原土壤有机质含量高光谱建模

为确定淮北平原砂姜黑土土壤有机质(SOM)最佳反演模型,探寻最佳特征波长筛选方法,提高模型预测精度。利用原始光谱进行倒数对数(Log(1/R))、标准正态变量变换(SNV)、去包络线(CR)、一阶微分(FDR)处理,采用稳定竞争性自适应重加权算法(sCARS)筛选特征变量,对比分析竞争性自适应重加权算法(CARS)、相关系数法(|r|≥0.47)和显著性水平法(p≤0.01)所得结果,建立SOM含量的偏最小二乘(PLSR)模型,并对比精度差异。结果表明:(1)全波段范围内,SOM含量与原始光谱呈极显著负相关,与Log(1/R)光谱呈极显著正相关,与SNV光谱相关性明显增强。CR和FDR光谱与SOM含量呈不同程度的正负相关性。(2)对比全波段,CARS和sCARS算法能够有效去除光谱冗余信息,筛选得到特征波段数目仅占全波段的1%~5%。筛选后模型精度更高,相对分析误差(RPD)均大于1.8。(3)相比于CARS算法,sCARS算法具备更好的稳定性和精确性。筛选到的特征波段主要分布在800~850、1850~1900、2050~2500 nm区域。(4)Log(1/R)-sCARS模型精度最佳,建模集和预测集的决定系数(R2)分别提升了0.08和0.28,RPD值为3.05,对SOM含量预测极好。

基于SCAR分子标记和倍性鉴定兰属花卉的研究

基于特异性片段扩增(SCAR)和倍性鉴定兰属花卉,为杂交育种亲本选配奠定基础,以兰属花卉的27个大花蕙兰和31个国兰品种为试材,通过SCAR分子标记和流式细胞仪进行亲缘关系和倍性检测,构建系统进化树。SCAR分子标记构建的系统进化树结果显示:58个品种聚为3支,第1支包括8个大花蕙兰品种(福娘、523、红袍等)和16个国兰品种(碧龙红素、汗血宝马、出水芙蓉等),第2支为11个大花蕙兰品种(金玉满堂、日本香兰、红双喜等)和6个国兰品种(一代天骄、春兰麻壳素、如意素荷等),第3支包括8个大花蕙兰品种(616、黄金岁月、杨贵妃等)和9个国兰品种(碧龙奇莲、西蜀道光、大雪素等);倍性鉴定结果显示:27个大花蕙兰品种中有7个二倍体(日本樱花、绿翡翠、梦境等)、16个三倍体(黄金岁月、蝶影、日本香兰等)、4个四倍体(英雄、523、福娘等)。部分大花蕙兰和国兰品种亲缘关系较近,可用作杂交育种亲本,但由于大花蕙兰倍性较为丰富。因此,大花蕙兰作为杂交育种亲本,选配时需进行倍性鉴定。综上,SCAR分子标记能高效、全面和精准鉴别兰属花卉间的亲缘关系,结合倍性鉴定,可为高效杂交育种奠定基础。

阿拉尔棉田烟粉虱捕食性天敌的SCAR分子标记检测

为了明确阿拉尔垦区棉田烟粉虱种群隐种以及烟粉虱捕食性天敌种类,采集阿拉尔十二团棉田中的烟粉虱及烟粉虱捕食性天敌,利用B-F/B-R、Q-F/Q-R特异性引物检测烟粉虱隐种类型,利用烟粉虱TB-F94585/TB-R 94585特异性引物检测捕食性天敌肠道内是否存在烟粉虱DNA。结果表明:新疆阿拉尔垦区烟粉虱为MED隐种,初步发现烟粉虱的捕食天敌共有4目5科,包括多异瓢虫成虫、多异瓢虫幼虫、东亚小花蝽、中华草蛉幼虫、黄褐新园蛛、灌木新园蛛、直伸肖蛸。利用SCAR分子标记技术可以快速检测阿拉尔垦区烟粉虱捕食性天敌种类。