【Objective】To investigate the effect of serum containing Yifa Compound(YC) on the growth of hair papilla cells.【Methods】Hair papilla cells were cultured in vitro.After subculture for 4～6 generations,hair papilla cells at logarithmic growth phase were cultured with YC-containing serum(at the concentrations of 1.25,2.5 and 5.0?g/L respectively) and blank serum respectively.After the culturing,the proliferation curve of hair papilla cells was drafted by methylthiazolyltetrazolium(MTT) assay and their growth was observed under scanning electron microscope.【Results】Optical density of hair papilla cells was higher at logarithmic growth phase while lower at growth lag phase in high-dose YC-containing serum group than that in blank serum group.Moderate-and high-dose YC-containing serum promoted the agglutinating radial growth.【Conclusion】Serum containing YC can promote the proliferation of hair papilla cells.

To investigate the expression of androgen receptor (AR) in affected area of androgenetic alopecia (AA) and the relationship between its expression and various syndromes of traditional Chinese medicine (TCM). [Me- thods ]Eighty cases of AA were classified as damp-heat syndrome of spleen and stomach and deficiency syndrome of liver and kidney. Expression level of AR in the alopecia scalp and non-alopecia scalp of the cases was detected with immunohistochemical method and was compared with 32 healthy volunteers.In the cases of AA, AR expression was positive in the granulocyte layer, spinous layer and basal layer of epiderm as well as in the fibroblast, vascular endothelial cells, nerve tissue, arrectores pilorum, sweat gland, sebaceous gland, hair root sheath and hair papilla of dermis;but it was negative in horny layer of epiderm, subcutaneous fat, internal root sheath, hair shaft cells and hair matrocytes. The total AR-positive rate and the AR-positive rate in epiderm, sweat gland, sebaceous gland, hair root sheath and hair papilla were higher in the alopecia scalp of the cases than those in non-alopecia scalp of the cases and in healthy volunteers (P0.05).[Conclusion]AR is related to the pathogenesis of AA and the local abnormal AR content may be one of the pathogenic factors of AA. Pathological feature and AR expression differ with the syndrome patterns: AR expression rate is higher in AA with damp-heat of spleen and stomach than those with deficiency of liver and kidney.

[Objective] To investigate the effects of Yangyin Kangdu Powder (YKP) on peripheral blood T-lymphocyte subsets levels in X-ray-irradiated mice. [Methods] Forty-five mice were randomized into normal control, model and YKP groups. Except the normal group, the rats in model and YKP groups were irradiated with 6Gy X-ray to establish models with acute radiation-induced injury. Normal saline was given to the model group while YKP in the dosage of 10 g?kg-1?d-1 was orally administered to YKP group for one week. The changes of CD4 and CD8 T-lymphocyte subsets levels were detected by flow cytometry. [Results] Plasma levels of CD4 and CD8 and the ratio of CD4/CD8 decreased in the model group, the difference being significant as compared with the normal control group (P

【Objective】To search an effective therapy for total alopecia (TA) and universal alopecia (UA) .【Methods】Nineteen cases of TA and twelve of UA were treated with combined therapy: irradiation with high-performanceelectromagnetic wave equipment, acupoint injection on Zusanli point (ST36) , external application of Yifa Tincture B andhormones, internal medication of hormones and washing with Xianglian solution.【Results】Among 31 cases of TA andUA, 17 were cured, 6 were markedly effective, 5 effective and 3 ineffective, the total effective rate being 90.3%.【Conclusion】Combined treatment with Chinese and western medicine has a certain effect for TA and UA.

Objective To observe the influence of dexamethasone on the gene expression profiling of PBMCs from patients with internal heat due to Yin deficiency-type SLE. Methods The PBMCs from 3 patients with internal heat due to Yin deficiency-type SLE were incubated with dexamethasone of 10-6 g/L for 48 hours followed by the detection of gene expression profiling of PBMCs by a GeneChip. Results The incubation with dexamethasone upregulated the expressions of 85 genes and downregulated those of 126 genes in PBMCs from the SLE patients. Conclusions Dexamethasone can regulate the expressions of numerous clusters of genes in PBMCs from patients with internal heat due to Yin deficiency-type SLE, and functional proteins encoded by these genes are located at various organelles and regions of cells. Not all the results of gene regulation by dexamethasone favor the relief of SLE. To study the regulation of genes may be beneficial to the elucidation of mechanisms underlying the therapeutic effect of dexamethasone on SLE.

OBJECTIVETo investigate serum cortisol level and glucocorticoid receptors (GR) mRNA expression in peripheral blood mononuclear cells (PBMCs) in patients with severe alopecia areata and liver-kidney deficiency syndrome and their involvement in the pathogenesis of severe alopecia areata.

METHODSIn 32 patients with severe alopecia areata, serum cortisol levels were measured by chemiluminescence assay and GR mRNA expression in the PBMCs was detected using reverse transcription real-time fluorescence quantitative PCR before and after treatment, with 20 normal subjects serving as the controls.

RESULTSSerum cortisol level showed no significant difference between the cases and the normal controls (P>0.05). The expression of GR mRNA in the PBMCs was significantly lower in the patients than in the normal controls (P<0.05). The expression of GR mRNA was even lower after treatments in patients with alopecia areata (P<0.01).

CONCLUSIONSGC-GR disorder exists in severe alopecia areata. A decreased GR mRNA expression in the PBMCs can be involved in the pathogenesis of severe alopecia areata, and such pathological changes at the receptor and genetic levels might also serve as the microscopic basis of liver-kidbey deficiency syndrome in severe alopecia areata.

Adult ; Alopecia Areata ; blood ; Case-Control Studies ; Diagnosis, Differential ; Female ; Humans ; Hydrocortisone ; blood ; Leukocytes, Mononuclear ; metabolism ; Male ; Medicine, Chinese Traditional ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Receptors, Glucocorticoid ; genetics ; metabolism ; Young Adult

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 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.