Atopic dermatitis （AD） is a common pruritic and chronic inflammatory dermatosis in clinical practice and is one of the diseases responding specifically to traditional Chinese medicine （TCM）. With the launch of biological agents and small molecule drugs and the development and implementation of guidelines of diagnosis and treatment， clinical pathways of treatment of moderate to severe AD， and consensus on the whole-process management of AD， the clinical efficacy of moderate to severe AD has been significantly improved. However， there are still many unmet clinical needs that require more effective methods to meet. In response to the Opinions of the CPC Central Committee and the State Council on Facilitating the Inheritance， Innovation， and Development of Traditional Chinese Medicine and the spirit of the National Conference on TCM， the China Association of Chinese Medicine organized more than 20 experts in TCM dermatology， Western medicine dermatology， interdisciplinary fields， and industries to discuss the difficulties and advantages of TCM in the treatment of AD. TCM treatment for AD can not only improve rash and relieve itching but also solve many concomitant syndromes. The abundant external treatment methods of TCM have advantages for different special populations and rash characteristics. The concept of treating disease before its onset in TCM is in line with the chronic disease management mode of prevention and treatment of atopic march and prevention of recurrence. In addition， TCM therapy can reduce the use of topical glucocorticoids and has good safety. Regarding the comorbidity of AD， equal emphasis on TCM and Western medicine and multidisciplinary joint treatment should be advocated to achieve maximum benefit for patients. The exchange of TCM and Western medicine has clarified the positioning and advantages of TCM intervention in AD， providing guidance for clinical and scientific research.

BACKGROUND:Currently,electrospun nanofibers,which are biomimetic materials of natural extracellular matrix and contain a three-dimensional network of interconnected pores,have been successfully used as scaffolds for various tissue regeneration,but are still faced with the challenge of extending the biomaterials into three-dimensional structures to reproduce the physiological,chemical as well as mechanical properties of the tissue microenvironment. OBJECTIVE:To summarize the process and principles of electrostatic spinning and to explore the applications of the resulting electrospun nanofibers in tissue regeneration of skin,blood vessels,nerves,bone,cartilage and tendons/ligaments. METHODS:With"electrospinning,electrospun nanofibers,electrospun nanofiber scaffolds,tissue regeneration"as the Chinese and English search terms,Google Academic Database,PubMed,and CNKI were searched,and finally 88 articles were included for review. RESULTS AND CONCLUSION:(1)The electrospun nanofibers are a natural fibrous extracellular matrix mimetic material and contain a three-dimensional network of interconnected pores that have been successfully used as scaffolds for a variety of tissue regeneration applications.(2)Several papers have described the great potential of electrospun nanofiber scaffolds applied to the regeneration of skin,blood vessels,nerves,bones,cartilage and tendons/ligaments,providing a solid theoretical basis for its final application in clinical disease treatment,or for its transformation into practical products to enter the market.(3)However,the current research results are mostly based on cell experimental research results in vitro,and whether it can be finally applied to human body still needs clinical verification.(4)At present,many kinds of electrospun products for various clinical needs have been commercialized in and outside China,indicating that the research field of electrospun nanofiber scaffolds for soft and hard tissue regeneration has great research value and application potential.

Objective A serum proteomic approach was used to explore the targets of action of Peitu Qingxin Granules(composed of Rhizoma Atractylodis Macrocephalae,Forsythiae Fructus,Imperatae Rhizoma,Pseudostellariae Radix,etc.)in the treatment of atopic dermatitis.Methods Five patients with atopic dermatitis were selected and treated with Peitu Qingxin Granules for 12 weeks,and five healthy volunteers were used as controls.The clinical core evaluation indexes of atopic dermatitis patients after treatment,including Eczema Area and Severity Index/Scoring Atopic Dermatitis(EASI/SCORAD),Pruritus Score,Patient-Oriented Eczema Measure(POEM),and quality of life index,were assessed.Serum samples were examined using data-independent acquisition-mass spectrometry(DIA-MS)technology,and serum differential proteins between atopic dermatitis patients and healthy people,as well as serum differential proteins in atopic dermatitis patients before and after treatment with Peitu Qingxin Granules were screened according to P＜0.05 and Fold Change>1.2.GO function enrichment analysis and KEGG pathway enrichment analysis were performed on the differential proteins.Results(1)Compared with the pre-treatment period,the clinical core evaluation indexes of patients with atopic dermatitis,including the EASI/SCORAD,Pruritus Score,POEM,and quality-of-life index,were significantly improved after treatment,and the differences were all statistically significant(P＜0.05,P＜0.01).(2)A total of 28 differential proteins were analyzed in the healthy control group and atopic dermatitis group,of which 12 proteins expressions were increased and 16 proteins were decreased,including ALAD(δ-aminolevulinic acid dehydrogenase),LTA4H(leukotriene A-4 hydrolase),CA1(carbonic anhydrase 1),F11(coagulation factor XI),and LCP1(lymphocyte cytoplasmic protein 1),etc..The main signaling pathways involved are PI3K-AKT signaling pathway,lipids and atherosclerosis,ECM-receptor interaction,platelet activation,NF-κB signaling pathway,and neutrophil extracellular trap formation.(3)A total of 12 different proteins were analyzed in atopic dermatitis patients before and after treatment with Peitu Qingxin Granules,of which 8 proteins were increased and 4 proteins were decreased,including ALAD,FGA(fibrinogen α-chain),IGHV3-64D,and IGHV3-38.They were mainly involved in signaling pathways such as lipids and atherosclerosis,complement pathway,Staphylococcus aureus infection,NF-κB signaling pathway,fluid shear stress and atherosclerosis.(4)The expressions of three protein targets including ALAD,FGA and IGHV3-64D,were significantly down-regulated in patients with atopic dermatitis and significantly up-regulated after treatment with Peitu Qingxin Granules.Conclusion The differentially expressed proteins ALAD,FGA and IGHV3-64D may be the action targets of Peitu Qingxin Granules in the treatment of atopic dermatitis,which lays the foundation for further experimental validation.

Objective To establish a mouse model of spleen deficiency and dampness stagnation syndrome combining atopic dermatitis(AD)and explore the feasibility of modeling by comparing 2,4-dinitrochlorobenzene(DNCB)-induced atopic dermatitis model of mouse,"external dampness+improper diet+irrigation of senna"-induced spleen deficiency and dampness stagnation syndrome model of mouse,as well as both in combination of model mouse.Methods The construction of a mouse(Balb/c)with spleen deficiency and dampness stagnation syndrome was explored by using the method of"external dampness+improper diet+irrigation of senna",and then DNCB was applied to induce the AD-like lesions in Balb/c mice to establish a mouse model of spleen deficiency and dampness stagnation syndrome combining atopic dermatitis.The general condition and body weight of mice in each group were observed,and the symptoms of spleen deficiency and dampness were scored.The severity of AD was evaluated by comparing the skin lesion degree,EASI score,transcutaneous water loss value(TEWL),spleen index and thymus index.The levels of creatinine,glucose,total cholesterol,triglyceride,gastrin,and amylase were measured.Results(1)During the modeling period of spleen deficiency and dampness stagnation syndrome,compared with the normal group,spleen deficiency and dampness stagnation syndrome group,spleen deficiency and dampness stagnation syndrome combined with atopic dermatitis group showed obesity,listlessness,filthy and greasy hair,diarrhea,and poor cleanliness around the anal.After combining with the application of the atopic dermatitis model,the body weight of the mice in atopic dermatitis group(P＜0.001),spleen deficiency and dampness stagnation syndrome group(P＜0.05)and spleen deficiency and dampness stagnation syndrome combined with atopic dermatitis group(P＜0.001)decreased sharply compared with the normal group.(2)Compared with the atopic dermatitis group,the degree of skin lesions,EASI score(P＜0.05)and TEWL(P＞0.05)were higher in the spleen deficiency and dampness stagnation syndrome combined with atopic dermatitis group.(3)Compared with the normal group,the spleen index of the atopic dermatitis group increased(P＜0.001)and the thymus index decreased(P＜0.001).Compared with the atopic dermatitis group,the spleen index(P＞0.05)and thymus index(P＜0.05)of the spleen deficiency and dampness stagnation syndrome combined with atopic dermatitis group decreased.(4)The results of serum biochemical indexes showed that compared with the normal group,the levels of creatinine(P＜0.01),glucose(P＜0.001),total cholesterol(P＞0.05),triglyceride(P＞0.05)and gastrin(P＜0.001)in the spleen deficiency and dampness stagnation syndrome group were increased,and the level of amylase was decreased(P＜0.01).Compared with the atopic dermatitis group,the levels of creatinine(P＞0.05),glucose(P＜0.05),total cholesterol(P＞0.05),triglyceride(P＞0.05),gastrin(P＜0.001)increased and the level of amylase decreased(P＞0.05).Conclusion A mouse model of spleen deficiency and dampness stagnation syndrome combining atopic dermatitis,which was induced by the combination of DNCB and"external dampness+improper diet+irrigation of senna",can not only show obvious TCM indications of spleen deficiency and dampness syndrome,but also show the characteristics of AD.This model can be used as a reliable animal model of combination of disease and syndrome.It provides reference for further study on pathological mechanism,pharmacodynamic evaluation and pharmacological mechanism of spleen deficiency and dampness stagnation syndrome combining atopic dermatitis.

The study aimed to evaluate the safety and function of poly(lactic-acid-co-ε-caprolactone) (PLCL)/fibrinogen nanofibers (P/F-Ns), and provide theoretical basis for the clinical application. The surface morphology, mechanical properties, the hydrophilicity and the fibrinogen content of P/F-Ns were tested by scanning electron microscope, the material testing machine, the contact angle meter and the microplate reader, respectively. The cell adhesion, proliferation and ligament remodeling genes expression of Hig-82 cells on P/F-Ns were conducted through cell counting kit-8 (CCK-8) and real-time quantitative PCR analyses, respectively. The results showed that with the increase of the fibrinogen content, the pore sizes and hydrophilicity of three P/F-Ns increased, but the mechanical properties decreased. Cell adhesion and proliferation tests showed that P/F-N-2 held the best ability to promote cell adhesion and proliferation. The ligament remodeling genes expressions of Hig-82 cells on P/F-N-1, P/F-N-2 and P/F-N-3 were all up-regulated compared to P/F-N-0 on days 3 and 7. All the three P/F-Ns containing fibrinogen (P/F-N-1, P/F-N-2 and P/F-N-3) had better biocompatibility compared to P/F-N-0, and could be efficiently applied to the reconstruction of anterior cruciate ligament.
Anterior Cruciate Ligament Reconstruction ; Cell Adhesion ; Fibrinogen ; Materials Testing ; Nanofibers

Objective:To evaluate the effect of danshensu on proliferation and apoptosis of M5-stimulated HaCaT cells (a psoriasis-like cell model) , and to explore its relationship with Yes-associated protein (YAP) expression.Methods:HaCaT cells were stimulated with M5, a mixture containing 10 μg/L interleukin (IL) -1α, IL-17, IL-22, tumor necrosis factor (TNF) -α and oncostatin M, for 48 hours to establish a psoriasis-like cell model. Then, the cell model was divided into several groups to be treated with 0 (control group) , 0.125, 0.25 and 0.5 mmol/L danshensu respectively, and HaCaT cells receiving no treatment served as the blank control group. Real-time quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of YAP respectively in these groups; methyl thiazolyl tetrazolium (MTT) assay was conducted to estimate the cellular proliferative activity after 24-, 48- and 72-hour treatment with danshensu, flow cytometry to evaluate the effect of danshensu on cell cycle and apoptosis, and Western blot analysis to determine expression of cell cycle-related proteins (cyclin A, cyclin B1, cyclin D, cyclin E) and apoptosis-related proteins (cleaved caspase-3, Bcl-2, BAX, p53 and p21) . One-way analysis of variance was used for comparing means in several groups, and least significant difference (LSD) - t test for multiple comparisons. Results:The mRNA and protein expression of YAP significantly differed among the blank control group, control group, 0.125-, 0.25- and 0.5-mmol/L danshensu groups (both P < 0.001) , so did the cellular proliferative activity at 24, 48 and 72 hours (all P < 0.001) . The 0.125-, 0.25- and 0.5-mmol/L danshensu groups all showed significantly decreased mRNA and protein expression of YAP (mRNA: 1.76 ± 0.04, 1.54 ± 0.05, 1.33 ± 0.05 respectively; protein: 1.78 ± 0.06, 1.49 ± 0.32, 1.27 ± 0.04 respectively) , and cellular proliferative activity at 48 hours (1.66 ± 0.04, 1.52 ± 0.02, 1.34 ± 0.04 respectively) compared with the control group (mRNA: 2.04 ± 0.04; protein: 2.10 ± 0.06; cellular proliferative activity: 1.82 ± 0.03; all P < 0.05) . Flow cytometry showed significant differences in the proportions of cells at G0/G1, S and G2/M phases as well as in the apoptosis rates among the above 5 groups (all P < 0.001) . Compared with the control group, the 0.125-, 0.25- and 0.5-mmol/L danshensu groups showed significantly higher proportions of cells at G0/G1 and G2/M phases, but lower proportions of cells at S phase (all P < 0.05) . Additionally, the apoptosis rates were significantly higher in the 0.25- and 0.5-mmol/L danshensu groups than in the control group (both P < 0.05) . Western blot analysis revealed significant differences in the expression of cell cycle-related proteins and apoptosis-related proteins among the above 5 groups (all P < 0.001) . Conclusion:Danshensu can inhibit the proliferation of the psoriasis-like cell model and promote its apoptosis, likely by suppressing YAP expression.

OBJECTIVE： To investigate the acute toxicity， long-term toxicity， skin irritation and anaphylaxis of Bee venom （BV） plastics， and to evaluate its preclinical safety. METHODS： The acute toxicity of BV plastics to rats was investigated after administration of high-dose， medium-dose and low-dose （144， 96， 48 mg/kg） of BV plastics. The long-term toxicity of BV plastics was investigated by continuous administration of high-dose， medium-dose and low-dose （72， 48， 24 mg/kg） of BV plastics for 28 days. The irritation of intact and damaged skin in rabbits with 8 mg/kg BV plastics was investigated by using the self-control method of left and right homologous body. The skin anaphylaxis of guinea pig were investigated after sensitized with 15 mg/kg BV plastics on the left back （on 0， 7th， 14th day） and stimulated with 15 mg/kg BV plastics on the right back. RESULTS： During the acute toxicity experiment with BV plastic，the weight of rats and the changes of viscera were normal，and there was no relevant toxic reaction. Long-term toxicity test results showed that no significant pathological changes were observed at 24 h after the last administration； the spleen index  of rats in BV low-dose group， testicular index in middle-dose group and epididymis index in high-dose and middle-dose groups were significantly increased， while PT in plasma of rats in BV medium-dose and low-dose groups was significantly prolonged （P＜0.05）. There were no abnormal changes in organ appearance， other organ index， coagulation index and blood biochemical index. All above indexes became normal at the end of 2-week recovery period. Skin irritation test showed that BV plastics could cause slight erythema and obvious scab on the skin of rabbits which along with little irritation on intact or damaged skin. Skin anaphylaxis test showed that BV plastics produced mild erythema in the skin of guinea pigs， belonging to light allergy. CONCLUSIONS： No acute or long-term toxicity is observed after transdermal administration of BV plastics， which is safe and only causes mild irritation and irritability to skin， indicating there is good safety of the plastic under experiment doses.

Objective:To investigate the sensitivity and the specificity of scorpions amplification refractory mutation system ( ARMS) in comparing with that of direct DNA sequencing in the detection of BRAF gene mutations in patients with papillary thyroid microcarcinoma.Methods:Direct sequencing and ARMS were used simultaneously to detect BRAF mutation status in 56 patients with PTMC.Results:BRAF mutations were identified in 46 cases with a mutation rate of 82.9%by ARMS,while in 18 cases with a mutation rate of 32.1%by direct sequencing.Besides,the sensitivity of ARMS was 100%and that of direct sequencing was 39.1%.There were significant differences of both mutation rate and sensitivity between two methods ( P<0.01 ).Conclusion: Compared to direct sequencing,ARMS gains a higher sensitivity in the detection of BRAF mutations in samples with tiny lesions.

BACKGROUND:The electrospinning technique has been used to prepare biological scaffolds to simulate nano-fiber structure of extracelular matrix; therefore, widespread attention has been paid to the electrospinning technique in the field of regenerative medicine and tissue engineering. OBJECTIVE: To review the articles about increasing electrospun nanofiber scaffold porosity, enlarging pore diameter, promoting cel infiltration with related technologies, in order to discover the most practical and economical technology. METHODS:The first author retrieved CNKI database, Wanfang database and PubMed with the keywords of “cel infiltration, 3D scaffold, electrospinning” in Chinese and English, respectively. Literature retrieval period was from January 2004 to October 2014. RESULTS AND CONCLUSION:Electrospinning technology is the most effective method for preparation of nanofiber scaffolds. Electrospinning scaffolds as tissue engineering scaffolds have become an issue of concern in the basic research year by year. However, the internal nano-scale pore of nanofiber scaffolds limits the cels to grow on the surface, so recent research has been focused on highly porous three-dimensional structure which can promote the permeable growth of cels instead of two-dimensional scaffolds. Several techniques have been used, which go from the adjustment of materials and speed of electrospinning to the applications of various kinds of complicated machines. However, the existing researches are stil not mature and stable, the majority of which are applied onlyin vitro as cel implantation or subcutaneous implantation in smal animals. The above-mentioned methods stil need long-term comparative studies to confirm the feasibility in the tissue-engineered repair of organs.

BACKGROUND:Due to the much higher requirement of biocompatibility and anticoagulant of smal-diameter vascular grafts than those of large-diameter ones, in situ blood vessel regeneration occurs as a new research direction. OBJECTIVE:To summarize the recent research development of electrospun smal-diameter scaffolds and to explore the application of in situ blood vessel regeneration and the development tendency. METHODS:The first author retrieved China National Knowledge Infrastructure database, Wanfang data and ISI Web of Knowledge foreign database to retrieve literatures addressing the fabrication of electrospun smal-diameter nanofibrous vascular grafts, surface modification and mimicking extracel ular matrix, as wel as the evaluation of biocompatibility and security after grafting. RESULTS AND CONCLUSION:Electrospun smal-diameter nanofibrous vascular grafts have emerged as promising candidates in vascular tissue engineering. By using both natural and synthetic polymers, the scaffolds can achieve a good balance between mechanical property and biocompatibility. Meanwhile, the fabrication of multi-layered vascular scaffolds, functional surface modification and mimicking extracel ular matrix structural y and functional y are now becoming attractive research directions. However, at current stage, electrospun vascular scaffolds used clinical y are basical y formed by synthetic materials, which have limited biocompatibility and anticoagulant activity. In this case, more efforts should be paid to find an optimal ratio between natural and synthetic materials for the improvement of biocompatibility and anticoagulant ability of smal-diameter vascular grafts.