Combined ozone hydrotherapy for atopic dermatitis:evaluation of efficacy and detection of interleukin-4 and nerve growth factor levels in peripheral blood from patients before and after treatment / 中华皮肤科杂志

Objective To evaluate the clinical efficacy and safety of combined ozone hydrotherapy for the treatment of atopic dermatitis(AD). Methods A total of 60 patients with moderate or severe AD aged from 6 to 65 years were enrolled, and randomly and equally divided into a test group and a control group. Both the two groups were treated with oral levocetirizine capsules 5 mg once a day, topical tacrolimus ointment twice a day, and topical moisturizers. The test group was additionally treated with ozone hydrotherapy 3- 5 times every week. The treatment lasted 2 weeks. The severity scoring of atopic dermatitis (SCORAD) score, visual analog scale (VAS) score, dermatology life quality index (DLQI) or children′s dermatology life quality index (CDLQI) score were assessed before and after the treatment, and compared between the two groups. Enzyme?linked immunosorbent assay(ELISA) was performed to measure the levels of interleukin?4(IL?4)and nerve growth factor(NGF)in peripheral blood from the patients before and after the treatment. Results After 2?week treatment, the SCORAD scores, VAS scores and DLQI/CDLQI scores significantly decreased from 42.13 ± 16.03, 7.14 ± 2.12 and 14.92 ± 5.94 before the treatment to 27.3 ± 11.01, 2.23 ± 1.31 and 9.69 ± 4.17 respectively in the test group(all P0.05). Conclusion Combined ozone hydrotherapy can effectively and safely improve the condition of patients with AD, likely by decreasing the levels of IL?4 in peripheral blood.

Propranolol in the treatment of problematic infantile hemangiomas / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the short-term efficacy and safety of propranolol for problematic infantile hemangiomas.@*METHODS@#Oral propranolol was administered to 68 infants with heamngiomas diagnosed by clinical evaluation and adjuvant examination at 1.0~2.0 mg per kilogram of body weight per day, divided to 2 or 3 times. The patients revisited once a month. The changes of the tumor size, texture, and color were monitored and recorded at a regular interval.The adverse effects after medication were observed and managed accordingly.The short-term results were evaluated using a 4-grade system.@*RESULTS@#All the 68 infants were followed up for 3-13 months, except that 1 infants combined with other diseases and 4 withdrew.The overall response was Scale 1 in 8 infants, Scale II in 13, Scale III in 29, and Scale IV in 13. No serious adverse effects were seen, but none cured entirely as well.@*CONCLUSION@#Oral propranolol is safe and effective for infantile heamngioma with good short-term result. It could be used as the primary drug for problematic infantile hemangiomas at the rapid growth stage of hemangiomas.

Inhibitive effect of triptolide on invasiveness of human fibrosarcoma cells by downregulating matrix metalloproteinase-9 expression

OBJECTIVE@#To explore the molecular mechanisms of antitumor properties of triptolide, a bioactive component isolated from the Chinese herb Tripterygium wolfordii Hook F.@*METHODS@#Human fibrosarcoma HT-1080 cells were treated with different doses of triptolide for 72 h. Then the expression and activity of matrix metalloproteinase (MMP)-2 and -9 were measured and the invasiveness of triptolide-treated HT-1080 cells was compared with that of anti-MMP-9-treated HT-1080 cells.@*RESULTS@#18 nmol/L triptolide inhibited the gene expression and activity of MMP-9, but not those of MMP-2, in HT-1080 cells. In addition, both 18 nmol/L triptolide and 3 μg/mL anti-MMP-9 significantly reduced the invasive potential of HT-1080 cells, by about 50% and 35%, respectively, compared with the control. Whereas there was no significant difference between the effect of 18 nmol/L triptolide and that of anti-MMP-9 on invasive potential of HT-1080 cells.@*CONCLUSIONS@#These data suggest that triptolide inhibits tumor cell invasion partly by reducing MMP-9 gene expression and activity.

RUNX3 gene methylation in a cutaneous malignant melanoma cell line A375 / 中华皮肤科杂志

Objective To investigate the relationship between the methylation of CpG island of RUNX3 gene promoter and its expression in a human cutaneous malignant melanoma cell line A375, and to assess the role of RUNX3 gene methylation in the pathogenesis of human cutaneous malignant melanoma. Methods Cultured A375 cells were treated with various concentrations (0, 1, 5, 10, 20 μmol/l) of 5-azacyti-dine for 24 or 72 hours followed by another 5 days of culture. Then, methylation-specific PCR (MSP) was performed to evaluate the methylation status of RUNX3 promoter region, and Western-blot analysis to detect the protein expression of RUNX3 in A375 cells. Results The RUNX3 gene promoter region was hypermethylated in untreated A375 cells, along with the absence of protein expression of RUNX3. However, after the treatment with 5-azacytidine, the promoter region of RUNX3 gene was demethylated partly, and the expression of RUNX3 protein was restored in A375 cells. Further, the expression intensity was directly correlated with the concentration of 5-azacytidine. Conclusions The promoter hypermethylation of RUNX3 gene may be related to the silencing of RUNX3 gene expression in A375 cells, whereas 5-azacytidine can cause the demethylation of RUNX3 gene, reactivate the gene which has been inactivated by the promoter hypermethylation, and finally induce the re-expression of RUNX3 protein.

Triptolide evaluates DNA methylation level of matrix metalloproteinase 9 gene in human fibrosarcoma HT-1080 cells / 中国中药杂志

<p><b>OBJECTIVE</b>The effect of triptolide on the DNA methylation level of MMP-9 gene and the mRNA expression of tissue inhibitors of met-alloproteinases (TIMPs) were examined in human fibrosarcoma HT-1080 cells to explore the molecular mechanisms involved in the anticancer activity of triptolide.</p><p><b>METHOD</b>HT-1080 cells were cultured in MEM containing 10% newborn calf serum and 1% penicillin-streptomycin. Triptolide was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 1 goL-1 and stored at -20 degrees C. Triptolide was freshly diluted with culture medium perior to use and directly added to cell cultures at the indicated concentration, and incubated for 72 hours at 37 degrees C in a humidified atmosphere with 5% CO2, with changes of reagents every 24 hours. Methylation specific PCR (MSP)was applied to assess the methylation status of MMP-9 gene promoter, and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was employed to measure the mRNA expression of tissue inhibitors of metalloproteinases (TIMPs) in human fibrosarcoma HT-1080 cells after 72 hours of treatment with 6 nmol x L(-1), 12 nmol x L(-1) or 18 nmol x L(-1) triptolide, respectively.</p><p><b>RESULTS</b>The methylation index of MMP-9 gene promoter was statistically elevated in HT-1080 cells after 72 hours of treatment with 18 nmol L(-1) triptolide, compared with those in controls (0.61 +/- 0.10 vs 0.39 +/- 0.10, P < 0.05), while no significant difference was noted between 6 nmol x L(-1) or 12 nmol x L(-1) triptolide treated HT-1080 cells and controls (0.40 +/- 0.15 vs 0.39 +/- 0.10, 0.46 +/- 0.20 vs 0.39 +/- 0.10, respectively, both P > 0.05). The mRNA expression of TIMP-1, -2, -3 or -4 was not significantly changed in HT-1080 cells after 72 hours of treatment with the indicated concentrations of triptolide, respectively compared with those in controls (all P > 0.05).</p><p><b>CONCLUSION</b>The results demonstrated that triptolide upregulates the methylation level of MMP-9 gene in HT-1080 cells in vitro.</p>

Effect of phorbol-12-myristate-13-acetate on cyclooxygenase-2 expression in human HaCaT keratinocytes / 中华皮肤科杂志

Objective To investigate the effect of phorbol-12-myristate-13-acetate(PMA)on cyclooxygenase-2(COX-2) mRNA and protein expression in cultured human HaCaT keratinocytes,and the mechanism for cytotoxity of PMA against keratinocytes.MethodsRT-PCR and Westem blot were utilized to detect the expression of COX-2 mRNA and protein in cultured HaCaT ells at 24 hours after the treatment with various concentrations of PMA (0.1,1.0,10 mg/L).ResultsWithout any treatment,there was no or a weak expression df COX-2 mRNA and protein in HaCaT cells;incubation witll PMA resulted in the induction of the expression of COX-2 in HaCaT cells.The expression levels of COX-2 mRNA and protein in 10 mg/L PMA-pretreated HaCaT cells were significantly higher than those in 1.0 mg/L PMA-pretreated HaCaT cells,which was in turn higher than that in 0.1 mg/L PMA-pretreated cells and untreated cells;the difrerence was statistically significant (all P＜0.01).Conclusion These results suggest that PMA may be involved in keratinocyte tumorigenesis by upregulating he expression of COX-2 as well as synthesis and release of prostaglandin in keratinocytes.

THE DISTRIBUTION OF INTRAMUSCULAR NERVESIN DIFFERENT SKELETAL MUSCLES OF THE RABBIT / 解剖学报

Objective To investigate the distribution of intramuscular nerves in the different skeletal muscles of the rabbit. Methods Muscle architecture and modified sihler's neural staining methods were used. Results The nerve branches of innervating flat M.pectoralis major arose from the anterior pectoral and posterior pectoral nerve.The former innervated cross fibers mainly,it penetrated the middle of muscle belly and formed a “U-Shaped” nerve ansa.The latter innervated oblique fibers mainly.There were many anastomoses between them.The nerve of pennate M.plantaris derived from tibial nerve.After entering muscle,the nerve trunk gradually divided into many primary branches toward medial and lateral fibers,these branches then subdivided into numerous arboroid second and terminal branches toward all of muscle fibres;The nerve of innervating spindle M.extensor digitorum longus came from N.fibularis communis.Two extramuscular nerve trunk were seen.Superior trunk mainly innervated those muscle fibers of inserting in the second toe,Inferior one mainly distributed to the fibers of rest within this muscle.Conclusion The spatial arrangement of the muscle fibres were related to distribution of the intramuscular nerves;Walking of intramuscular nerve branches had two patterns which run perpendicular and/or parallel to muscle fasciculi.