Objective To investigate the effects of IL-15 eukaryotic expression plasmid on the Ag-specific immune responses induced by HPV 16 E7 vaccine in mice.Methods An eukaryotic expression plasmid encoding IL-15 gene coding region was constructed and named as pcDNA3.1IL-15.Female BALB/c mice were injected intramuscularly with pcDNA3.1-IL-15 and HPV16 E7 gene vaccine.After the immunization,the concentration of serum IFN-? were tested.Single-cell suspensions of splenocytes were prepared from mice and were re-stimulated with HPV16 E7 protein.Then,the T cell proliferation assay was measured by the MTT method.Results The concentration of serum IFN-? in mice after co-injection with pcDNA3.1-IL-15 and pcDNA3.1-E7 was raised to 414.1pg/ml,which was significantly higher than that of mice co-injected with pcDNA3.1 and pcDNA3.1-E7 or mice injected with pcDNA3.1-E7 only(P

Objective To study the role of calcitonin gene-related peptide(CGRP)and vasoactive intestinal peptide(VIP)in the pathogenesis of alopecia areata(AA).Methods Radioimmunoassay(RIA)was used to measure the levels of CGRP and VIP in plasma from30patients with AA and20normal controls.Immunohistochemistry was employed to detect the expression of CGRP and VIP in lesions of21patients with AA and16normal scalps.Results①The plasma levels of CGRP in progressing stage of AA(142.63?67.95pg/mL)were significantly lower than those in stable stage of AA(197.33?67.15pg/mL)and in normal controls(188.40?72.95pg/mL).②The plasma levels of VIP in progressing stage of AA(105.94?55.42pg/mL)were significantly lower than those in stable stage of AA(156.86?47.37pg/mL)and in normal controls(176.44?84.70pg/mL).③The expression of CGRP and VIP was significanly decreased in lesions of AA than that in normal scalps.Conclusion These findings indicate that CGRP and VIP may play a role in the pathogenesis of alopecia areata.

Objective To report a case of toxic epidermal necrolysis associated with geotrichosis due to Geotrichum silvicola. Methods The exudates from the body surface, blood and urine of the patient were examined by microscopy and simultaneously inoculated onto the Sabouraud dextrose agar (SDA) medium. The isolate was examined by microscopy, PCR which amplified the D1/D2 domain of 26S rDNA, and gene sequencing. Homologous sequences were searched in the GenBank/EMBL/DDBJ/PD nucleotide sequence library, and the genetic relationship was analyzed with the genealogical tree. Results Microscopy of pus from the abscess on the dorsa of left hand revealed a lot of spores and a few hyphae, which were not observed in the blood or urine specimens. Meanwhile, whitish colonies were grown in all the three successive cultures of blood and urine specimens, rather than the exudates on the body surface. After itraconazole and garlicin were administered for one week, both microscopic exam and fungus culture were negative. Microscopic exam of the isolate showed arthrospores arranged in chains, budding spores and a few of hyphae. It was found that there was a one-base difference between our isolate (Hebei-1) and the isolate from kerion -like eruptions (Changzheng-1), and a four-base difference between our isolate and the reference Geotrichum silvicola strain as well, in the D1/D2 domain of 26S rDNA. This isolate was identified to be most close to Changzheng-1 in the phylogenetic tree. Conclusion The patient with toxic epidermal necrolysis is associated with geotrichosis due to Geotrichum silvicola.

Objective To report a case of kerion caused by Geotrichum in China. Methods A 9 year-old-boy had kerion-form lesion on his scalp with swollen posterior auricular lymph nodes, and did not show other definite underlying disease. The pathogenic fungus was identified according to culture, scanning electron microscopy, biochemical tests and DNA sequencing. The hair infection test was performed and the infected hairs were examined by scanning electron microscope. Animal test confirmed the pathogenicity of the fungus. Results The fungal colonies were the same when the tissue cultures were repeated. The colonies showed milky white to yellowish in color. The hyphae could be identified at the periphery on Sabouraud′s agar culture at 27 ℃, which were moist and smooth on the surface at 37 ℃. Under microscope, there were many rectangular arthrospores, round or oval spores with or without buddings, as well as branched hyphae. The isolated fungus was identified as a Geotrichum silvicola by culture, scanning electron microscope, biochemical test and DNA sequencing. The patient′s condition was improved markedly after treatment of terbinafine for 4 weeks. Conclusions This is the first case report of kerion caused by Geotrichum in China, and terbinafine is effective.

Objective To study expression and significance of keratin 17 and 19 in psoriatic lesion.Method The expression of keratin 17 and 19 in 30 psoriatic lesion and 10 normal skin was measured by immunohistochemistry method.Results The expression level of keratin 17 in the psoriatic lesion was higher than that in the normal skin,the expression level of keratin 19 in the psoriatic lesion was lower than that in the normal skin,there were significant differences in the expression of keratin 17 and 19 between them(P ＜0.05).The optical density level of keratin 17 in the psoriatic lesion was obviously raised compared with the normal skin(5.81 ± 1.42 vs.0,P＜ 0.01).The optical density level of keratin 19 in the psoriatic lesion was obviously decreased compared with the normal skin(0.49 ±0.03 vs.2.03± 1.08,P＜0.05).The optical density level of keratin 17 and 19 showed negative correlation in the psoriatic lesion(r =-0.479,P＜ 0.01).Conclusion Keratin 17 and 19 may play a role in the pathogenesis of psoriasis.

Objective To determine the degree of damage to hairs by Geotrichum, and to compare the difference in infection duration and intensity by Geotrichum silvicola isolates from skin lesions and blood and Geotrichum candidumn isolates between hairs from different age groups. Methods In vitro hair perforation test was carried out on the hairs from healthy individuals of different age groups. Scanning electron microscopy was performed to observe hairs perforated by Geotrichum silvicola isolates from skin lesions and blood as well as Geotrichum candidumn isolates. Results Both Geotrichum silvicola and Geotrichum candidumn isolates could perforate hairs. The time taken to perorate hairs was significantly different between isolates of Geotrichum silvicola from skin lesions, blood and Geotrichum candidumn in every age groups (all P ＜ 0.05).The Geotrichum silvicola isolates from blood perforated hairs in the shortest time (range: 53 - 64 days, mean:58.07 ± 3.15 days), followed by those from skin lesions (range: 57 - 66 days, mean: 61.05 ± 2.55 days), and Geotrichum candidumn (range: 61 - 74 days, mean: 67.11 ± 3.78 days). The time taken to perforate hairs by Geotrichum significantly increased with age, and significant difference was observed between hairs from the four age groups, i.e., ＜ 2 years, 2 - 13 years, 14 - 19 yeas, ＞ 19 years, but not between the age group of ＜ 2 years and that of 2 - 13 years (P ＞ 0.05 ). As scanning electron microscopy showed, both Geotrichum silvicola and Geotrichum candidumn could damage hairs, and the degree of damage by Geotrichum silvicola was more severe than that by Geotrichum candidumn. Conclusions The damage to hairs by Geotrichum silvicola is earlier and more severe than that by Geotrichum candidumn. The younger the age, the more liable the hairs to be damaged by Geotrichum.

Objective To investigate the effects of 2-methoxyestradiol (2-ME) on the proliferation and apoptosis of a mouse malignant melanoma cell line B16,and to explore their mechanism.Methods B16 cells were cultured in vitro,and divided into a negative control group receiving no treatment and several intervention groups treated with 2-ME at final concentrations of 5,10,20,40 mmol/L,respectively.After different durations of treatment,inverted phase-contrast microscopy was conducted to observe the morphologic change of B16 cells,sulforhodamine B (SRB) assay to evaluate proliferative activity and to draw growth curve of B16 cells according to the absorbance value at 490 nm,flow cytometry to detect cell cycle and apoptosis,and reverse transcription PCR and real-time PCR were performed to measure the expressions of the apoptosis-inducing gene gadd45b and proto-oncogene c-myc.Results As repeated measures analysis of variance showed,there were significant differences in the inhibitory effect on B16 cell proliferation among different concentrations (5,10,20,40 mmol/L) and different treatment durations (24,48,72 hours) of 2-ME (F =1170.94,1843.04,respectively,both P ＜ 0.01),and there was a significant interaction effect between these concentrations and treatment durations (F =272.79,P ＜ 0.01).After 48-hour treatment with 2-ME at 10,20 and 40 mmol/L,the apoptosis rate of B16 cells was increased to (4.13 ± 1.12)％,(11.25 ± 2.380)％ and (19.46 ± 2.9)％ respectively,compared to (0.23 ± 0.5)％ in the negative control group (all P＜ 0.01); the proportion of B16 cells in G0/G1 phase was increased to (59.5 ± 5.6)％,(63.4 ± 8.2)％ and (70.8 ± 4.4)％ respectively,compared to (44.1 ± 3.4)％ in the negative control group.There was a significant difference in the proportion of B16 cells in G0/G1 phase among the negative control group and intervention groups (F =13.56,P ＜ 0.05).Moreover,the mRNA expression of gadd45b was significantly enhanced after 24-hour treatment with 2-ME at concentrations of 20 and 40 mmol/L (both P＜ 0.01),while that of c-myc was significantly weakened after treatment with 2-ME at 10,20 and 40 mmol/L (all ＜ 0.05) compared with the negative control group.Conclusion 2-ME can inhibit the proliferation of B16 cells in vitro,upregulate the expression of gadd45b gene and downregulate the expression of C-myc gene.

Objective:To investigate specific cellular immune response to the HPV16E7 prophylactic vaccine in mice.Methods:BALB/c mice were randomly divided into 3 groups:experimental group (treated with pcDNA3.1-HPV16E7),control group Ⅰ(treated with pcDNA3.1) and control group Ⅱ(treated with N.S.).Mice were injected (i.m.) pcDNA3.1-HPV16E7,pcDNA3.1 and N.S. one time per week,respectively.After three immunization,the blood samples from eye sockets and the supernatant cultured of spleen cells were taken for measurement IFN-? and the number of CD4 +?CD8 +T-lymphocyte by ELISA and FACS assay.Antigen-specific splenocyte proliferation assay in vitro was detected by MTT method.Results:The splenocytes actively proliferated,the number of CD4 +T lymphocyte and the quantitation of IFN-? in spleen and serum in the experimental group were significantly higher than the control group Ⅰ and Ⅱ.Conclusion:The pcDNA3.1-HPV16E7 DNA vaccine can induce specific cellular immune response in BALB/c mice.

OBJECTIVE:To observe effects and safety of Kangfuxin solution combined with intense pulsed light in the treat-ment of rosacea. METHODS:A total of 50 rosacea patients in our hospital during May 2014-Jun. 2016 were divided into control group(25 cases)and observation group(25 cases)according to random number table. Based on oral administration of Metronida-zole tablets,control group received intense pulsed light. Observation group was additionally given Kangfuxin solution for local wet compress after 4 to 6 layers of gauze saturated with liquid,5-10 min,qn. Both groups received treatment for 4 weeks. Clinical effi-cacies,as well as symptom score and DLQI score were compared between 2 groups before and after treatment,and the occurrence of ADR was recorded. RESULTS:The response rate of observation group was 92.0%,which was significantly higher than 64.0%of control group,with statistical significance (P<0.05). Before treatment,there was no statistical significance in erythema,pap-ules,pustules,itching,telangiectasia score and total score,DLQI score with before treatment(P>0.05). After treatment,erythe-ma,papules,pustules,itching,telangiectasia score and total score,DLQI score of 2 groups were decreased significantly,and the observation group was significantly lower than the control group,with statistical significance(P<0.05). The incidence of ADR in observation group was 16.0%,which was significantly lower than 40.0% in control group,with statistical significance(P<0.05). CONCLUSIONS:Kangfuxin solution combined with intense pulsed light show significant efficacy for rosacea,and can effectively improve erythema,papules,pustules,itching and telangiectasia,and improve the quality of life with good safety.

Adolescent ; Adult ; Child ; Deubiquitinating Enzyme CYLD ; Female ; Humans ; Male ; Neoplastic Syndromes, Hereditary ; genetics ; Pedigree ; Skin Neoplasms ; genetics ; Tumor Suppressor Proteins ; genetics ; Young Adult