The Effects of hydrogen peroxide(H2o2) on the ultrastructure, cellular pea-meability barrier and 3H-TdR-DNA leakage of C. albicans were studied. Damage of the cell, especially leakage of cytoplasm were found under transmission electron microscope. Morphological changes and small hole like structures were also found under scanning electron microscope. Radioactivity assay of the leakage of 3H-TdR-DNA leakage of C. albicans further suggested that the 3H-TdR-DNA leakage of C. albicans was directly due to the damage of cellular permeability barrier. Negative correlation (r=+0.8392--0.9915)between the extent of 3H-TdR-DNA leakage and the number of surviving colony-forming units was found. This indicated that the damage of cellular permeability barrier and leakage of DNA of C. albicans caused by H2o2 was one of the most importent causes of cell killing.

OBJECTIVE: To summarize the advance in research on a variety of data processing methods of optimizing drug preparation process. METHODS: On the basis of data processing methods of single index, this paper compares, analyzes and summarizes a variety of data processing methods in relevant literature. RESULTS: Multiple indicator data processing method, such as analysis of variance-comprehensive weighted of multi-index, multiple regression analysis-response surface methodology, artificial neural network, and multi-dimensional space triangle area, has been widely used in the optimization of drug preparation process in a certain scope. CONCLUSION: Analysis of variance-comprehensive weighted of multi-index, multiple regression analysis-response surface methodology, artificial neural networks, and multidimensional spatial triangular area can reveal the principles among multi-factors of multi-levels, thus can provide reference for optimizing drug preparation technology.

Obtaining soil bacterial DNA of good quality is a key step in soil bacterial ecology study.A quick, efficient,sensitive and stably method of DNA extraction from soil were established by combining strongpoints of two kits(Soilmaster kit and DNA IQ~(TM)kit).In addition,the 16S rDNA gene and T-RFLP(Terminal restriction fragment length polymorphism)were used in the analysis of soil bacterial community diversity and the result show that T-RFLP is a powerful tool for bacterial community study.

The antifungal mechanism of the TS67(Penicillum sp.) producing bioactive metabolites was investigated in this paper. The test indicated that mycelia growth, spores forming and spores germination of Bipolaris maydis and Fusarium oxysporum can be inhibitied by metabolites, after 50% fermentation broth acting 120 hours, the inhibition rate of hyphal growth can reach about 77.78%, 70.30%, the inhibition rate of spores forming also arrived at 58.8%, 73.5%; under 50% fermentation broth acting 12 hours, the inhibition rate of spores germination were up to 78.3%, 62.0%, the mycelia under activity metabolites conditoins also showed surface nodulation and growing point expanding irregularly, protoplasmic condesing, etc. Based on all these results, we may draw a conclusion that activity metabolites maybe target at fungal cell wall.

Objective To investigate the application of ultrasound visualization in instantly evaluation and supplement therapeu-tics in the treatment of uterine fibroids and adenomyosis with high intensity focused ultrasound (HIFU ) .Methods 57 patients with 67 uterine fibroids and 31 patients with 41 adenomyosis were treated with JC-200 HIFU treatment system and monitored the blood flow change in the lesion with B-ultrasound .Evaluated the curative effect with ablation ratio and ablation ratio after supplement therapeutics .Results The average ablation ratio of 57 uterine fibroids was(84 .6 ± 16 .1)% and the increased to(87 .0 ± 10 .7)% af-ter supplement therapeutics to 9 lesions with blood flow in the border of all .The changes were no significance(P>0 .05) .The aver-age ablation ratio of 31 adenomyosis was(62 ± 22 .7)% and increased to(74 ± 14 .7)% after supplement therapeutics to 11 lesions with blood flow in the border of all .The changes were statically significance(P<0 .05) .Conclusion Ultrasound visualization could be used to evaluate the area and extent of ablation with HIFU therapy ,it can clear lesions remaining parts and guiding the supple-ment therapeutics to improve the ablation ratio .Ultrasound visualization provided an evidence of therapeutics in the early period .

Objective To investigate the relationship of Toll-like receptors (TLRs) 2 and 4 with the occurrence of chloasma.Methods Peripheral blood samples were collected from 40 patients with chloasma and 40 healthy human controls,and skin samples were also collected from the lesions of 10 of the patients and normal skin of 10 of the healthy controls.Real time (RT)-PCR was performed to measure the mRNA expressions of TLR2 and TLR4 in skin lesions and blood samples.An immunohistochemical test was conducted to observe the expressions of TLR2 and TLR4 in skin lesions.Statistical analysis was carried out by t test.Results The expressions of TLR2 and TLR4 mRNAs were both significantly higher in skin lesions of the patients than in normal skin of the controls (9.72 ± 2.93 vs.5.10 ± 2.69,t =3.67,P＜ 0.01; 9.52 ± 2.88 vs.4.77 ± 1.90,t =4.36,P＜ 0.01),while no significant difference was found in the mRNA expressions of TLR2 or TLR4 in peripheral blood between the patients and controls (both P ＞ 0.05).As the immunohistochemical test revealed,TLR2 was absent in both the epidermis and vascular endothelial cells in 6 normal control skin samples,weakly expressed in the basal layer of the epidermis but absent in vascular endothelial cells in 4 normal skin samples,and no TLR4 expression was observed in either the epidermis or vascular endothelial cells in these control skin samples.Among the 10 skin samples from chloasma lesions,3 showed TLR2 expression in the whole epidermis,7 in both basal cell layer and prickle cell layer but not in vascular endothelial cells in the superficial dermal layer,all showed strong TLR4 expression in the basal cell layer and weak TLR4 expression in the prickle cell layer,and 3 exhibited TLR4 expression in vascular endothelial cells in the superficial dermal layer.Conclusion TLR-mediated immune responses in local skin might be related to the occurrence of chloasma.

ObjectiveTo seek experimental evidence of epidermal barrier dysfunction in psoriasis,and to provide a basis for adjuvant therapy of psoriasis.MethodsPhysiometric methods were used to determine the value of sebum content,transepidermal water loss(TEWL) and water content of stratum corneum in 60 patients with psoriasis and 48 normal human controls.The ultrastructure of lamellar bodies was observed with transmission electron microscopy,and the expression of acid ceramidase in normal skin and psoriatic lesions was detected by using immunohistochemical techniques.ResultsCompared with the normal skin,TEWL value was increased(P ＜ 0.01),but water content of stratum corneum decreased(P ＜ 0.01 ) in psoriatic lesions,and sebum content was similar between normal skin and psoriatic lesions.As electron microscopy showed,lamellar bodies in keratinocytes were reduced in number with a disorganized arrangement and irregular size in psoriatic lesions.The expression of acid ceramidase also decreased in psoriatic epidermis.Conclusions The function of epidermal barrier in psoriasis is impaired,and to restore epidermal barrier function and enhance hydration may serve as an important adjuvant therapy of psoriasis.

Objective To evaluate the effects of Prinsepia utilis Royle oil (PURO) on the synthesis of ceramide and expression of acid ceramidase N-acylsphingosine amidohydrolase 1 (ASH1),and to explore the mechanisms underlying its moisturizing and skin barrier-repairing effects.Methods Keratinocytes from human foreskin tissue were classified into 2 groups to be cultured in keratinocyte-serum free medium (K-SFM) with or without the presence of PURO.Enzyme linked immunosorbent assay (ELISA) was performed to measure the level of ceramide in the culture supernatant of keratinocytes at 0,3,8,24 and 48 hours.The back of nude mice was divided into 4 areas,i.e.,test area,matrix area,blank control area and negative control area.Acetone and ether were used to destroy the epidermal barrier in the test,matrix,and blank control areas,then,the former 2 areas were topically treated with emulsions containing 1％ PURO and matrix,respectively,and the blank control area remained untreated.The epidermal barrier remained intact and untreated in the negative control area.Noninvasive methods were used to determine transepidermal water loss (TEWL),epidermal moisture content and skin lipid content in these areas on day 0,1,3,and 7.Skin tissue was obtained from these areas on day 0 and 7 followed by an immunohistochemical study for the quantification of ASH1 expression.Results The level of supernatant ceramide increased with time in the PURO-treated keratinocytes,which was significantly higher at 24 hours and 48 hours than at 0 hour (1.3817 ± 0.100 and 1.3737 ± 0.047 vs.0.7630 ± 0.143,both P ＜ 0.05).The supernatant ceramide was also elevated in the PURO-treated keratinocytes compared with untreated keratinocytes at 24 and 48 hours (both P ＜ 0.05).Noninvasive skin tests showed a gradual decrease in the TEWL,but an increase in the epidermal moisture content and skin lipid content with time in the 3 epidermal barrier-destroyed areas.As far as the test area was concerned,TEWL value was significantly lower on day 3 and 7 than on day 0 (10.85 ± 0.64 and 8.01 ± 0.58 vs.12.65 ± 0.71,both P ＜ 0.05),while a significant increment was observed in the skin lipid content on day 3 and 7 compared with day 0 (29.14 ± 0.40 and 31.30 ± 0.88 vs.27.02 ± 0.65,both P ＜ 0.05),as well as in the epidermal moisture content on day 1,3 and 7 compared with day 0 (13.98 ± 0.28,15.00 ± 0.38 and 15.86 ± 0.18 vs.11.74 ± 0.62,all P＜ 0.05).On day 7,there was a statistical decline in TEWL value,but an elevation in epidermal moisture content,skin lipid content and ASH1 expression in the test area compared with the matrix area and blank control area (all P ＜ 0.05).Also,the expression of ASH1 was upregulated on day 7 compared with day 0 in the 3 barrier-destroyed areas (all P ＜ 0.05).Conclusion PURO may exert skin-moisturizing and barrier-repairing effects by enhancing the synthesis of ceramide and expression of acid ceramidase ASH1.