Objective To investigate the relevant risk factors for,clinical manifestation and treatment responses of symmetric acral keratoderma in 9 patients.Methods Clinical data were collected from 9 outpatients with symmetric acral keratoderma.Skin lesions were photographed,tissue specimens were obtained from the lesions and subjected to fungal and histopathological examinations.The patients were given topical treatment.Results There were 8 males and 1 female in these patients with an average age of 28.2 years and age at onset of 24.6 years.The disease duration varied from 2 months to 16 years.The lesions were predominantly distributed on the dorsum of both hands and wrists symmetrically and circularly.Ankles were the third popular site involved.Generally,the lesions were aggravated in summer,spontaneously cured in winter.Most of the patients had a history of occupational contact with plastics or rubber.Some patients suffered from ichthyosis vulgaris.Microscopic examination for fungal element was negative.Histopathology revealed epidermal hyperkeratosis,acanthosis and mild papillomatosis.The urea and Vitamin E cream mixed with clobetasol propionate cream at a ratio of 1 ∶ 1 proved to be effective.Conclusions Ichthyosis vulgaris may be a risk factor for symmetric acral keratoderma.The whitening of lesions after short exposure to water may be a kind of physical phenomenon with the involvement of water and air.Topical steroids may serve as the first-line treatment choice for symmetric acral keratoderma.

Objective To investigate the histological changes of normal human skin after fixed irradiation with multiple-pulse fractional Er:YAG laser.Methods After hair removal,the upper arms of 7 healthy volunteers were consecutively irradiated with multiple-pulse fractional Er:YAG laser.Tissue samples were resected from the irradiated skin at 1,24,48,72 hours,on day 5,7,15,and 30 after the irradiation.Hematoxylin and eosin(HE)staining and Masson staining were performed to observe the histological changes and collagen proliferation respectively.Immunohistochemical staining was used to detect the expression of heat shock protein(HSP)70 and 47.Results The consecutive irradiation with multi-pulse fractional Er:YAG laser generated an array of tapering microscopic treatment zones(MTZs)of gasification and ablation in the upper arm skin.After the irradiation,inflammation developed in the microscopic lesions with the epithelization of epidermal cells within 7 days; local dermal collagen was renewed and remodelled during the 7th to 30th day.HSP70 expression peaked as early as 48 hours after the treatment and maintained until the 7th day,while the high expression of HSP47 persisted from the 15th to 30th day after irradiation.Conclusion The fixed,multiplepulse and fractional Er:YAG laser irradiation can reach the deep dermis,and induce the local proliferation of dermal collagen.

Objective To study the impact of HIV-1 V2 L175P mutation on the binding capability of anti-V3 neutralizing antibodies to HIV-1.Methods A series of eukaryotic cell expression plasmids were used to concatenate wild type and mutant env gene of HIV-1 and green fluorescent protein(GFP)gene.The recombinant plasmids were transfected into 293T cells to express HIV-1 gp120 protein on the surface of cells.The successfully transfected cells were screened by GFP florescence marker.Immunostaining and dual fluorescence flow cytometry were performed to test the binding affinity of several common V3 region specific neutralizing antibodies to wild type or mutant gp120 proteins.Results The mean fluorescence intensity(MFI)of mutant gp120-expressing 293T cells were significantly higher than that of negative control cells(expressing GFP).Flow cytometry showed that the curve for mutant gp120-expressing 293T cells was obviously different in shape and peak from that for the negative control,while most parts of the curve for the wild type gp120-expressing 293T cells overlapped with those for the negative control.Conclusion The V2 region mutation may increase the sensitivity of HIV-1 to the neutralization by V3 region specific antibodies.

Objective To analyze the composition of and drug resistance in bacteria isolated from the lesions of patients with squamous cell carcinoma of the face and head.Methods Lesional tissue or discharges were obtained from 246 patients with squamous cell carcinoma of the face and head,and subjected to conventional bacterial culture.The isolated bacteria were identified by VITEK TWO automated microbiology system.Antimicrobial susceptibility testing was carried out by Kirby-bauer method.WHONET 5.3 software was utilized for statistical analysis.Results Totally,294 bacterial strains were isolated,including 168 Gram-negative bacteria (57.1％)and 126 Gram-positive bacteria(42.9％).The bacterial isolates were predominated by Staphylococcus aureus(21.4％),followed by Escherichia coli(20.4％),Staphylococcus epidermidis(18.4％),Klebsiellapneumoniae(15.4％)and Pseudomonas aeruginosa(9.5％).The prevalence was 40％,26.7％,42.9％ and 55.6％ respectively for extended spectrum β lactamases-producing E.coli and K.pneumoniae,methicillinresistant staphylococcus aureus(MRSA)and methicillin-resistant coagulase-negative S.epidermidis(MRCNS)respectively.P.aeruginosa,E.coli and K.pneumoniae were highly susceptible to imipenem and meropenem,and favorably sensitive to β-lactam and β-1actamase inhibitor combination.No resistance was observed for vancomycin,teicoplanin or linezolid in staphylococci.Conclusions The bacterial isolates from squamous cell carcinoma tissue on the head and neck are predominated by conditional pathogenic bacteria,and the proportion of Gram-negative bacteria is higher than that of Gram-positive bacteria.These isolates seem to be highly resistant to common antibiotics.

Objective To investigate the changes of IL-8 in peripheral blood of patients with pustular psoriasis after acitretin treatment.Methods Dual antibody sandwich enzyme-linked immunosorbent assay (ELISA)was performed to measure the IL-8 levels in the peripheral blood of 30 patients with pustular psoriasis and 30 patients with psoriasis vulgaris before and after treatment,and in 30 normal human controls.The relationship between the IL-8 level and disease severity was assessed for patients with pustular psoriasis.Results After acitretin treatment,the condition was improved greatly in patients with pustular psoriasis,together with a significant decrease in the level of IL-8 in the peripheral blood(57.07 ± 12.02 pg/ml vs.96.84 ± 14.68 pg/ml,P ＜ 0.05).Conclusion IL-8 may be involved in the therapeutic mechanism of acitretin in pustular psoriasis.

Objective To investigate the expression of granulocyte macrophage-colony stimulating factor(GM-CSF)in skin lesions and sera of patients with pustular psoriasis and psoriasis vulgaris.Methods Tissue specimens were obtained from the lesions of 15 patients with pustular psoriasis,15 patients with psoriasis vulgaris and 15 normal human controls.Immunohistochemistry and dual antibody sandwich enzyme-linked immunosorbent assay(ELISA)were carried out to detect the levels of GM-CSF in the tissue and serum specimens from the patients and normal human controls,respectively.Results Significantly higher levels of GM-CSF were observed in the tissue and serum specimens from patients with pustular psoriasis and psoriasis vulgaris compared with the normal controls(all P ＜ 0.01),as well as in those from the patients with pustular psoriasis compared with the patients with psoriasis vulgaris(both P ＜ 0.01).Conclusion GM-CSF may be involved in the pathogenesis of psoriasis.

Two cases of nevus of Ota resistant to multiple sessions of Q-switched laser treatment are reported.Both of the patients were females,who developed beige or blue patches on the left side of the face (zygomatic region,cheek)several months after birth.No conjunctiva or sclera was involved.The lesions spread to the left retroauricular region and posterior neck during pregnancy in one patient.Histopathological examination of the lesions showed scattered melanocytes and melanophages in lower dermis and subcutaneous adipose tissue as well as apparent proliferation of collagen.Immunostaining for CD34 showed no abnormality in the distribution of vascular endothelial cells in these tissues.No obvious improvement was observed in either of the patient after more than 10 sessions(12 sessions in 1 case,and 13 sessions in the other case)of treatments with Q-switched alexandrite(755 nm)or Q-switched Nd:YAG laser(1064 nm).The immediate response of lesions to laser irradiation was unusual.Purpura or errhysis was absent in the lesions after repetitive irradiation with high-energy laser,but both were visible in the adjacent normal skin even after irradiation with low-energy laser.

Objective To investigate the expression of tumor necrosis factor alpha(TNF-α)and its relationship with infiltrating lymphocytes in lichen planus(LP).Methods Tissue specimens were obtained from the lesions of 60 patients with LP and normal skin of 20 human controls.Immunohistochemical SP method was used to detect the expression of TNF-α,and infiltrating lymphocytes were counted in TNF-α-positive tissue sections.Results TNF-α was expressed in 72％ of the LP specimens but in none of the control specimens(P ＜ 0.01).Positive staining for TNF-α was mainly located in the membrane of prickle cells,cytoplasm or membrane of dermal infiltrating lymphocytes.The expression of TNF-α in LP was uncorrelated with age,sex or disease course(all P ＞ 0.05),but was positively correlated with infiltrating lymphocyte number (rs =0.47,P ＜ 0.01).Conclusion TNF-α seems to play a certain role in the pathogenesis of LP.

Objective To retrospectively analyze the therapeutic effect of wide surgical excision combined with low-dose adjuvant interferon-alpha 2b on cutaneous malignant melanoma.Methods A total of 32 patients with cutaneous malignant melanoma received wide surgical excision after preoperative examination and staging.The excisions were performed with a margin measuring 1-2 cm from the visible lesions or biopsy scars.Surgical modalities included direct suture after excision(4 patients),dactylolysis or toe amputation(6 patients),free skin grafting(15 patients),random skin flap transfer(3 patients)and pedicle skin flap transfer(4 patients).Lymph nodes were selectively dissected in 9 patients with regional transfer of lymph nodes,and inguinal lymph nodes were cleared away in 2 patients.One week after the operation,patients received adjuvant therapy with subcutaneous injection of interferon-alpha 2b(3 million IU,thrice per week)for one to three years.Results Preoperative tumor staging revealed 21 cases of cutaneous malignant melanoma at stage Ⅱ,and 11 cases at stage Ⅲ.The excisions healed by the first stage in all the patients.Up to June 2011,2 patients had been lost to follow up,5 patients with stage Ⅲ melanoma had died.Survival was observed in all of the 4 patients receiving 1-year follow up,12 of 13 patients receiving 1-3 year follow up,5 of 7 patients receiving 3-5 year follow up,and 4 of 6 receiving 5-year follow up.Of the 25 surviving patients,regional lymph node metastasis was observed in 8 patients,which developed within 2 years after the operation in 2 patients.The adjuvant therapy with interferon-alpha 2b lasted 3 years in 8 patients,and more than 1 year in 11 patients.Side effects were mild.Conclusion Wide surgical excision plus low-dose interferon-alpha 2b is effective for the treatment of stage Ⅱ and Ⅲ cutaneous malignant melanoma with lower local recurrence and higher survival rate.

Objective To investigate DJ-1 expression and protective effect against H2O2-induced oxidative stress in primary human melanocytes.Methods The expression and location of DJ-1 in primary melanocytes were identified by immunofluorescence.After cultured melanocytes were exposed to H2O2 for 24 hours,modified MTT assay was used to assess the proliferation of cells and to choose the suitable concentration of H2O2 for the following experiment.Western blot was used to detect D J-1 expression in melanocytes after being treated with 0.5 mmol/L H2O2 for 24 hours.Some melanocytes were divided into 3 groups to be reversely transfected with PBS(mock control group),non-targeting siRNA(negative control group)and DJ-1 targeting siRNA(DJ-1 group).Optical microscopy was utilized to observe the morphologic changes of transfected melanocytes.At 48 hours after the transfection,the melanocytes were stimulated with H2O2 for 24 hours.Subsequently,modified MTT assay,2′,7′-dichlorofluorescein diacetate(DCFH-DA)and annexin Ⅴ-fluorescein isothiocyanate/propidium iodide were used to determine cell viability,intracellular reactive oxygen species (ROS)level and apoptosis rate respectively.Results DJ-1 was expressed in both melanocyte nucleus and cytoplasm,and predominantly in the nucleus.H2O2 inhibited the cell viability in a dose dependent manner.After treatment with H2O2 of 0.5 mmol/L for 24 hours,the cell viability began to decrease in melanocytes with the expression level of DJ-1 being 2.23 times that in the untreated melanocytes(both P ＜ 0.05).Compared with the mock control group,the dendrites of melanocytes in DJ-1 group were obviously shortened with cytoplasm vacuolization.After 24-hour treatment with H2O2 of 0.5 mmol/L,the cell viability in the DJ-1 group dropped to 35％ of that in the mock control group(P ＜ 0.05),while the intracellular ROS fluorescence intensity( FI) and apoptosis rate were higher in the DJ-1 group than in the mock control group (902 ± 40 vs.529± 32,58q％ ± 6.1％ vs.30% ± 3.8％,both P ＜ 0.05).Conclusion DJ-1 can protect melanocytes against H2O2induced oxidative stress likely by decreasing intracellular ROS production and inhibiting ceU apoptosis.