Effect of danshensu on Yes-associated protein expression in as well as on proliferation and apoptosis of a psoriasis-like cell model / 中华皮肤科杂志

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.

Serum cortisol and peripheral blood mononuclear cell glucocorticoid receptor mRNA expression in severe alopecia areata with liver-kidney deficiency syndrome / 南方医科大学学报

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>In 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.</p><p><b>RESULTS</b>Serum 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).</p><p><b>CONCLUSIONS</b>GC-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.</p>

Influence of dexam ethasone on the gene expression profiling of peripheral blood mononuclear cells (PBMCs) from patients with internal heat due to Yin deficiency-type SLE / 中华皮肤科杂志

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.

Effect of Yifa Compound on the Growth of Hair Papilla Cells / 广州中医药大学学报

【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.

Effect of Yangyin Kangdu Powder on Peripheral Blood T-lymphocyte Subsets in X-ray-irradiated Mice / 广州中医药大学学报

[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

Pathological Feature and Expression of Androgen Receptor in Affected Area of Androgenetic Alopecia in Various Syndromes of Traditional Chinese Medicine / 广州中医药大学学报

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.

A Clinical Observation of Combined Treatment with Chinese and Western Medicine for Total Alopecia and Universal Alopecia / 广州中医药大学学报

【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.