Objective:To investigate the effect of astragaloside IV (AS-IV) on the secretion of stromal cell-derived factor-1α (SDF-1α) and CXC chemokine receptor 4 (CXCR4) by high glucose injured human umbilical vein endothelial cells (HUVECs), so as to lay a foundation for further study on AS-IV improving angiogenesis by regulating SDF-1 α/CXCR4 axis of endothelial cells.Methods:HUVECs were isolated and cultured from the umbilical vein of full-term healthy newborns and identified by von Willebrand factor (vWF) combined with 4-diamino-2-phenylindole (DAPI) nuclear staining. The obtained HUVECs was cultured in EGM-2 medium with 30 mmol/L glucose for 120 h to obtain high glucose damaged HUVECs. After intervention with different concentration gradients (25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L, 400 mg/L) AS-IV for 72 hours, the contents of SDF-1α and CXCR4 were detected by enzyme linked immunosorbent assay (ELISA) method to determine the best concentration of AS-IV. The supernatant of damaged HUVECs were collected at 6, 12, 24, 48 and 72 hours after intervention with the best concentration of AS-IV, and the contents of SDF-1α and CXCR4 were detected by ELISA method to determine the best action time of AS-IV. The damaged HUVECs was randomly divided into experimental group and control group, and the blank group was set up at the same time. The experimental group was treated with the best concentration of AS-IV and the best time, the control group and the blank group were treated with the same volume of phosphate buffered saline (PBS) solution, and the contents of SDF-1α and CXCR4 in each group were detected by ELISA method.Results:The vWF factor on the cell membrane was green fluorescence, and the nucleus was blue after DAPI staining. When the fusion image showed green fluorescence, HUVECs were identified by blue fluorescence. The expression of SDF-1α in damaged HUVECs was the best when treated with AS-IV of 100 mg/L for 24 hours (1 642.87 pg/ml), and the expression of CXCR4 in damaged HUVECs was the best when treated with AS-IV of 50 mg/L for 48 hours (8.44 ng/ml). Compared with the control group, the contents of SDF-1α and CXCR4 in the experimental group were significantly increased, and the difference was statistically significant ( P<0.05). While the contents of SDF-1α and CXCR4 in the experiment group were slightly less than those in the blank group and there was no statistically significant difference ( P>0.05). Conclusions:AS-IV can promote the expression of SDF-1α and CXCR4 in HUVECs damaged by high glucose to return to normal physiological level, so as to play the role of vascular repair and neovascularization.

Objective:To establish a safe, effective, simple, and more economically feasible method to obtain platelet-rich plasma (PRP).Methods:Whole blood was collected from 24 patients with atrophic acne scars on the face. For the preparation of PRP, a slight modification of Choukroun＇s method was used. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of platelet derived growth factor-β(PDGF-β), transforming growth factor-β1 (TGF-β1), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), isulin-like growth factor-1 (IGF-1) in normal plasma and PRP.Results:The average concentration of PDGF-β in PRP was (755.61±418.31) ng/L, which was higher than [(479.93±279.18) ng/L] in normal plasma ( t=3.479, P<0.01). The average concentration of TGF-β1 in PRP was (2267.00±1223.68), which was higher than [(1535.50±910.91) ng/L] in normal plasma ( t=7.082, P<0.01). The average concentration of EGF in PRP was (30.70±12.39) ng/L, which was higher than [(20.77±10.31) ng/L] in normal plasma ( t=6.899, P<0.01). The average concentration of VEGF in PRP was (25.42±17.69) ng/L, which was higher than [(12.01±7.77) ng/L] in normal plasma ( t=5.230, P<0.01). The average concentration of bFGF was (17.85±7.17) ng/L, which was higher than [(10.90±4.73) ng/L] in normal plasma ( t=6.050, P<0.01). The average concentration of IGF-1 was (201.22±36.80) ng/ml, which was higher than [(174.90±33.80) ng/ml] in normal plasma ( t=3.760, P<0.01). Conclusions:Compared to normal plasma, modified PRP contains higher levels of growth factors. The modified method is a reliable option for liquid PRP, especially applicable for invasive cosmetic laser surgery to promote wound repair.

Objective To investigate sun protection behavior among middle-aged and elderly women in Beijing.Methods The central (Xuanwumen district) and peripheral areas (Yanqing district) of Beijing city were selected for investigation from May to June in 2016.By random sampling and field investigation,interviewees completed questionnaires with the help of dermatologists.Results A total of 400 females aged 40-90 years were investigated,including 190 in the Xuanwumen district and 210 in the Yanqing district.Among all the subjects,the average duration of outdoor activities was 2 hours.Moreover,117 (29.2％) of the 400 interviewees had the active sun exposure habit,including 38 (18.1％) in the Yanqing district and 79 (41.6％) in the Xuanwumen district,and the proportion of subjects with the active sun exposure habit significantly differed between the two districts (x2 =26.582,P ＜ 0.001).In addition,67 (16.8％) interviewees had a sunburn history,and 130 (32.5％) were used to applying sunscreen.The proportion of sunscreen users was significantly higher in moderately or highly educated women (103 [45.2％]) than in lowly educated women (28/172 [16.3％],P ＜ 0.001).The proportion of sunscreen users in women with skin type Ⅲ (41.2％ [70/170]) was significantly higher than that in those with skin type Ⅳ (26.5％ [61/230],P =0.002).The scores for facial skin aging and dorsal hand skin aging were both significantly lower in sunscreen users than in non-users (Z =18.536,10.731,P =0.002,0.026,respectively).Conclusion The active sun exposure habit in the middle-aged and elderly women differs between the central and peripheral areas of Beijing,and the use of sunscreen is associated with education level.

Objectives: To explore weather oxygen uptake efficiency slope (OUES) may predict the prognosis in patients with idiopathic pulmonary arterial hypertension (IPAH). Methods: The consecutive newly diagnosed IPAH patients in our hospital from 2010-11 to 2015-06 were prospectively enrolled and regular follow-up study was conducted to record cardiovascular events (death and lung transplantation). Kaplan–Meier curve, uni- and multivariate Cox regression analysis were performed to assess the survival rate in relevant patients. Results: A total of 210 IPAH patients at the mean age of (32±10) years were finished cardiopulmonary exercise test (CPET) and received regular follow-up study including 159 female. There were 31 patients died and 1 received lung transplantation over 41 months follow-up period. OUES was positively related to peak oxygen uptake (VO2)/body weight (r=0.71, P<0.0001). Multivariate analysis demonstrated that OUESI and NT-proBNP could independently predict the prognosis of IPAH patients. The 5-year survival rate in patients with OUESI≤0.52 L/(min?m2) was lower than those with OUESI>0.52 L/(min?m2) (41.9% vs 89.8%), P<0.0001.Conclusion: OUES as a submaximal CPET parameter may well predict the prognosis in IPAH patients.

Objective To evaluate effects of fine particulate matter PM2.5 in ambient air on the proliferation,cell cycle and apoptosis of a human keratinocyte cell line HaCaT.Methods PM2.5 in hazefog episodes during the heating season was collected in Beijing from 2015 to 2016,and processed into PM2.5 suspensions.HaCaT cells were divided into several groups to be treated with culture medium alone (control group),PM2.5 suspensions at different concentrations of 100-400 mg/L (experiment groups,50-800 mg/L for observation of cellular morphology and analysis of cell proliferation) for 24 hours,or cell culture medium without cells or PM2.5 suspensions (blank group).Cellular morphological changes were observed under an inverted microscope.Cell counting kit-8 (CCK-8) assay was performed to determine cell survival rate,flow cytometry to determine the cell cycle distribution and detect cell apoptosis,and Western blot analysis to determine the protein expression of cyclin A2 and cyclin-dependent kinase1 (CDK1).Results Along with the increase of PM2.5 concentration,HaCaT cells lost their normal shape gradually,and the number of viable cells gradually decreased.Compared with the control group (100％ ± 4.95％),the 50-mg/L PM2.5 group showed no changes in cell survival rates (P ＞ 0.05),while the 100-,200-,400-and 800-mg/L PM2.5 group showed significantly lower survival rates (91.77％ ± 2.04％,80.01％ ± 1.57％,57.80％ ± 1.56％,21.98％ ± 0.86％,respectively,all P ＜ 0.05).Flow cytometry revealed that the 100-,200-and 400-mg/L PM2.5 groups showed gradually increased proportion of cells at S phase,but gradually decreased proportion of cells at G2/M phase compared with the control group (all P ＜ 0.05).As Western blot analysis showed,the protein expression of cyclin A2 and CDK1 significantly decreased in the 100-,200-and 400-mg/L PM2.5 groups compared with the control group,which was lowest in the 200-mg/L PM2.5 group(all P ＜ 0.05).In addition,the 100-,200-and 400-mg/L PM2.5 groups showed significantly higher total apoptosis rates (9.98％ ± 0.21％,12.56％ ± 0.74％,16.74％ ± 1.48％,respectively) compared with the control group (6.24％ ± 0.17％,all P ＜ 0.05).Conclusion PM2.5 can inhibit cell proliferation and promote apoptosis of HaCaT cells,likely by downregulating the expression of cyclin A2 and CDK1 and arresting HaCaT cells at S phase.

Objective To investigate the syndrome differentiation characteristics of children with atopic dermatitis(AD)and the immune imbalance status in children with different syndrome types of AD.Methods A total of 159 AD children and 100 normal control children were enrolled.The peripheral blood eosinophil(Eo)count was measured by impedance method,total serum immunoglobulin E(IgE)by immunoturbidimetric assay,and interferon-gamma(IFN-γ),interleukin-4(IL-4),interleukin-5(IL-5)and interleukin-17(IL-17)were measured by multiple microspheres flow immunofluorescence assay.Results Among 159 AD children,syndrome of heart-fire and spleen-deficiency was most commom,accounting for 38.4%,followed by syndrome of blood-deficiency and wind-dryness(22.0%),syndrome of heat accumulation in heart and spleen(20.1%)and syndrome of spleen-deficiency and dampness-accumulation(19.5%).Compared with normal control group,there was no significant difference in serum IFN-γ level among different syndrome types of AD.The levels of peripheral blood Eo,serum total IgE,IL-4 and IL-17 in AD with heart-fire and spleen-deficiency syndrome were significantly increased(P<0.05).The levels of peripheral blood Eo,IL-4,IL-5 and IL-17 in AD with blood-deficiency and wind-dryness syndrome were significantly increased(P<0.05).The levels of IL-4,IL-5 and IL-17 in AD with heat accumulation in heart and spleen syndrome were significantly increased(P<0.05).The levels of peripheral blood Eo and serum IL-4 in AD with spleen-deficiency and dampness-accumulation syndrome were significantly increased(P<0.05).Conclusion Heart-fire and spleen-deficiency syndrome is the most common type in children with AD,however,the main type under 3 years old is heat accumulation in heart and spleen syndrome.Th2/Th17 immune imbalance are the main pathogenesis in heart-fire and spleen-deficiency syndrome,blood-deficiency and wind-dryness syndrome and heat accumulation in heart and spleen syndrome,and Th2 immune imbalance is the main pathogenesis of spleen-deficiency and dampness-accumulation syndrome.

Objective:To investigate the effects of Astragaloside Ⅳ on the secretion of exosomes in human endothelial progenitor cells (EPCs) and the expression of microRNA (miRNA)-126 in exosomes.Methods:The umbilical cord blood from one healthy full-term newborn from the Department of Obstetrics and Gynecology of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine in 2019 was harvested for isolating mononuclear cells by density gradient centrifugation and cultured for 7 days. Morphological observation was performed during this period. Cells of the third passage were collected for identification by CD31 immunomagnetic bead sorting and double fluorescence staining. According to the random number table, the identified EPCs were divided into Astragaloside Ⅳ group and phosphate buffer solution (PBS) group. The cells in Astragaloside Ⅳ group were cultured with Astragaloside Ⅳ in final mass concentration of 100 mg/L for 24 hours, and the cells in PBS group were cultured with the same volume of PBS for 24 hours. After culture, the exosomes from the cell culture supernatant of the two groups were collected, and the expressions of characteristic markers of exosomes CD9, CD63, and CD81 were detected by Western blotting, the morphology of EPC exosomes (EPC-Exos) was observed under transmission electron microscope, and the particle size of EPC-Exos was detected by nanoparticle tracking analysis technique. The concentration of EPC-Exos was determined by dioctyl butyric acid method (the sample number was 3), and the expressions of miRNA-126-3p and miRNA-126-5p related to angiogenesis in EPC-Exos were determined by reverse transcription polymerase chain reaction (the sample number was 3). Data were statistically analyzed with independent sample t test. Results:(1) On the 4th day of culture, the cells began to adhere to the wall, and the multi-forms such as circle, fusiform, and strip appeared at the same time. On the 7th day of culture, the edge of the cells was clear and arranged like a paving stone, the central cells were round, and the surrounding cells were fusiform. (2) CD31 immunomagnetic beads sorting method identification showed that the membrane was stained with green fluorescence and the nucleus was stained with blue fluorescence. Double fluorescence staining method showed that the cells were orange-yellow. The cells were identified as EPCs. (3) After 24 hours of culture, the expressions of CD9, CD63, and CD81 in EPC-Exos were all positive, confirming that EPC-Exos were extracted successfully in this experiment. (4) After 24 hours of culture, the EPC-Exos of the two groups showed round membrane vesicles, and there was no significant difference in morphology. (5) After 24 hours of culture, the particle size of 98.7% EPC-Exos in Astragaloside Ⅳ group was 84.7 to 143.1 nm, and that of 98.0% EPC-Exos in PBS group was 88.7 to 123.5 nm. (6) After 24 hours of culture, the mass concentration of EPC-Exos in Astragaloside Ⅳ group was (310±5) μg/mL, which was significantly higher than (257±5) μg/mL in PBS group, t=13.369, P<0.01. (7) After 24 hours of culture, there were more miRNA-126-3p ( t=16.062, P<0.01) and miRNA-126-5p ( t=3.252, P<0.05) in EPC-Exos of Astragaloside Ⅳ group than in PBS group. Conclusions:Astragaloside Ⅳ can improve the function of human EPC secretory exosomes, and the secreted exosomes are loaded with miRNA-126.

Objective:To identify pathogenic genes in 3 cases of piebaldism, and to explore the genotype-phenotype relationships in piebaldism.Methods:Clinical data were collected from 3 patients with piebaldism and their parents at the Department of Dermatology, Henan Provincial People′s Hospital from January 2019 to December 2021. Peripheral blood samples were obtained from them and 100 unrelated healthy controls, and DNA was extracted. Whole-exome sequencing technology was used to screen genetic variation sites, and then Sanger sequencing was performed for verification. The deleteriousness of genetic variants was evaluated by using pathogenicity analysis software tools.Results:Case 1: a 23-year-old male patient presented with white patches on the forehead, chest, and abdomen for 23 years, and his parents had no similar symptoms; case 2: a 1-year- and 5-month-old male infant presented with white patches on the forehead and abdomen for 1 year, and his parents had no similar symptoms; case 3: a 6-year-old male child presented with white patches on the forehead and limbs for 6 years, and his parents had no similar clinical manifestations. Genetic testing showed that a missense mutation c.2033T>C (p.L678P) in exon 14 of the KIT gene, a splice site mutation c.2485-1G>C in exon 18 of the KIT gene, and a heterozygous missense mutation c.2346C>G (p.F782L) in exon 16 of the KIT gene were identified in the case 1, 2, 3 respectively, but no above mutations were identified in the patients′ parents or 100 unrelated healthy controls. The 3 genetic variants were all novel pathogenic mutations, and all were deleterious mutations.Conclusions:Three novel pathogenic mutations in the KIT gene were identified in the 3 cases of piebaldism, namely c.2033T>C (p.L678P), c.2485-1G>C, and c.2346C>G (p.F782L). It was further verified that the severity of piebaldism was closely related to the type and location of KIT gene mutations.

Background::Urticaria is a common skin disease characterized by episodes of wheals, and it has a negative effect on patients’ quality of life. Large-scale population-based epidemiological studies of urticaria are scarce in China. The aim of this survey was to determine the prevalence, clinical forms, and risk factors of urticaria in the Chinese population.Methods::This survey was conducted in 35 cities from 31 provinces, autonomous regions, and municipalities of China. Two to three communities in each city were selected in this investigation. Participants completed questionnaires and received dermatological examinations. We analyzed the prevalence, clinical forms, and risk factors of urticaria.Results::In total, 44,875 questionnaires were distributed and 41,041 valid questionnaires were collected (17,563 male and 23,478 female participants). The lifetime prevalence of urticaria was 7.30%, with 8.26% in female and 6.34% in male individuals ( P < 0.05). The point prevalence of urticaria was 0.75%, with 0.79% in female and 0.71% in male individuals ( P < 0.05). Concomitant angioedema was found in 6.16% of patients. Adults had a higher prevalence of urticaria than adolescents and children. Living in urban areas, exposure to pollutants, an anxious or depressed psychological status, a personal and family history of allergy, thyroid diseases, and Helicobacter pylori infection were associated with a higher prevalence of urticaria. Smoking was correlated with a reduced risk of urticaria. Conclusion::This study demonstrated that the lifetime prevalence of urticaria was 7.30% and the point prevalence was 0.75% in the Chinese population; women had a higher prevalence of urticaria than men. Various factors were correlated with urticaria.

A 1-year and 9-month-old male proband presented with clustered rice grain-sized flat smooth red papules on the face, trunk and limbs for 1.5 years, without fever, joint swelling, or pain. The proband′s sister aged 7 years ever experienced swelling and pain in the finger joints of both hands at the age of 2 years, and had intermittent fever and papules all over the body at the same time, and the papules gradually regressed with the subsidence of fever. The proband′s mother aged 27 years suffered from swelling and pain in the finger joints of both hands when she was young, gradually leading to finger deformities, and experienced intermittent knee swelling and pain at the age of 12 years without obvious skin lesions on the body. No abnormality was found in ophthalmological and systemic physical examinations of the 3 patients. Whole-exome sequencing showed that the proband, his sister and mother all had a heterozygous missense mutation c.1001G>A (p.R334Q) in exon 4 of the NOD2 gene. A diagnosis of Blau syndrome was made. The proband was treated with topical moisturizing cream all over the body; during the 52-week follow-up, no joint swelling and pain or eye symptoms were found in the proband, while erythema and depressed scars were observed on the face, trunk and limbs. The proband′s sister and mother were treated with subcutaneous injections of adalimumab at initial doses of 40 mg and 80 mg respectively, followed 1 week later by injections at 20 mg and 40 mg respectively, and then treated with injections at 20 mg and 40 mg respectively every 2 weeks; after 12-week treatment, the joint swelling and pain were markedly relieved in the proband′s sister and mother, and most skin lesions subsided in the proband′s sister; at week 52 during the follow-up, there was no joint swelling, pain or skin lesions in the proband′s sister, and there was no swelling or pain in the knee joints of the proband′s mother, while no improvement was observed in her finger deformities. During the treatment, no eye symptoms or adverse reactions were observed neither in the proband′s sister nor in his mother.