Objective To estimate the influences of N-acetyl cysteine (NAC) on a chemical hypoxiamimetic agent CoCl2 induced-injury to, and expressions of inflammatory factors by, an immortal human skin keratinocyte line HaCaT. Methods HaCaT cells were treated with CoCl2 of 2000 μmol/L for 4 hours to set up a chemical hypoxia-induced cell model of skin injury. NAC of various concentrations ( 1000, 2000, 3000 μmol/L)was used to pretreat HaCaT cells for 2 hours prior to the establishment of cell model. After these treatments,cell viability was detected by cell counting kit 8 (CCK-8), the levels of interleukin 6 and 8 (IL-6 and -8) and tumor necrosis factor α (TNF-α) in culture supernatant by ELISA kits, mitochondrial membrane potential (MMP) by rhodamine 123 (Rh123) staining and photofluorography, intracellular reduced glutathione (GSH)content by glutathione detection kit. Results An obvious decline was observed in HaCaT cell viability after pretreatment with various concentrations of NAC for 2 hours. The treatment with CoCl2 of 2000 μmol/L for 4 hours induced an elevation in the supernatant levels of IL-6, IL-8 and TNF-α and a decrease in GSH content and MMP, while the pretreatment with NAC for 2 hours retarded the CoCl2-induced increase in IL-6 and IL-8 levels as well as decrease in GSH content and MMP. Conclusion The reactive oxygen species (ROS) scavenger NAC can protect against CoCl2-induced injury to and inflammatory reaction in HaCaT cells, which may be associated with a decrement in oxidative stress.

Objective To investigate the expression and activation of extracellular signal-regulated kinase 1/2 (ERK 1/2), its upstream molecule, epidermal growth factor receptor (EGFR), and downstream transcription factor, Ets-like protein 1 (ELK-1), in lesions of psoriasis vulgaris, and to evaluate the relationship between ERK pathway and psoriasis vulgaris. Methods Tissue samples were obtained from the lesions of 40 patients with psoriasis vulgaris and normal skin of 20 normal human controls. Immunohistochemistry and Western blot were performed to detect the expressions of phosphorylated ERK1/2, EGFR and ELK-1 in the tissue samples.Results As immunohistochemistry showed, the integrated optical density (IOD) of p-ERK1/2, p-EGFR and p-ELK-1 was 269.85 ± 57.96, 136.88 ± 30.33 and 237.61 ± 56.29 respectively in the psoriatic lesions, significantly higher than that in the normal controls ( 140.24 ± 24.42, 110.66 ± 28.99 and 119.04 ± 21.99, respectively, all P ＜ 0.05). A positive correlation was observed between the expression of p-EGFR and p-ERK1/2(r = 0.57, P ＜ 0.05) and between that of p-ERK1/2 and p-ELK-1 (r=0.72,P＜0.05) in psoriatic lesions.Conclusion The enhanced signal transduction through phosphorylated EGFR→ERK1/2→ELK-1 pathway may play a certain role in the pathophysiological process of psoriasis vulgaris.

Objective To profile the omp1 genotypes of Chlamydia trachomatis (Ct) in patients with nongonococcal urethritis (cervicitis) in Guangzhou region. Methods Swab samples were obtained from the urethra of males and cervix of females in clinical settings of venereology and gynecology as well as at outreach sites for the prevention and control of sexually transmitted diseases (STDs). DNA was extracted from the swabs and nested PCR was performed to amplify the variable domain (VD) 1 - 3 of omp1 gene of Ct followed by gene sequencing. The genotypes of Ct were determined based on the amino acid mutation in VD 1 - 2 of omp1 gene. Results Totally, 1208 swabs were collected. Of them, 132 were Ct positive, and 130 positive samples underwent genotyping. Ten ompl genotypes were determined in total, including serotype E (38, 29.23%), D (25, 19.23%), J(24, 18.46%), F(21, 16.15%), G(7, 5.38%), H(5, 3.85%), K(5, 3.85%), B(2, 1.54%), Ja (2, 1.54%), I (1, 0.77%). E, D, J and F were the dominant type of Ct in this region, and amounted to 83% of all the Ct isolates. Mutations were observed within VD 1 and 2 of omp1 gene in serotype D, B and K.Serotypes were undetermined for Ct in 2 patients with mixed infection. Conclusions In Guangzhou region, E,D, F and J are the predominant genotypes of Ct, and amount to 83% of all the Ct isolates. Ct serotype B is also observed in the urethra of males and cervix of females in this region.

A 20-year-old male patient presented with myalgia of upper limbs and myasthenia of extremities for more than 1 month. Physical examination showed diffuse erythema on the cheeks, upper eyelids, upper chest, neck and dorsa of the hands. The myodynamia of the proximal and distal muscles of upper and lower extremities was grade Ⅳ, Ⅴ, Ⅲ and Ⅴ respectively. Laboratory examinations revealed that the serum levels of creatine kinase, CK-MB and lactate dehydrogenase were 2103 U/L, 83 U/L and 489 U/L respectively, which were all above the normal range. Electromyogram revealed myopathic abnormality and normal nerve conduction velocity. Histopathology of gastrocnemius muscle showed hypertrophy and swelling of muscle fibers, disappearance or fuzziness of transverse striation, and intermuscular lymphoid cell infiltration. A biopsy of the skin lesion from the upper chest showed liquefaction degeneration of and colloid bodies in basal cell layer, perivascular lymphoid cell infiltration in the dermis. A diagnosis of dermatomyositis was established based on the clinical and laboratory findings. After management with intravenous prednisolone 80 mg once daily and symptomatic treatment for 4 weeks, the myodynamia of upper limbs was improved, serum levels of creatine kinase,CK-MB and lactate dehydrogenase reached the normal ranges. However, the myodynamia of lower limbs progressively deteriorated with the emergence of paresthesia. Enhanced MRI scan showed a tumor in the vertebral canal at the level of thoracic vertebra 11 to 12. A spherical encapsulated tumor measuring 3 cm in diameter was surgically removed. The tumor was diagnosed as paraganglioma in vertebral canal according to pathological and immunohistochemical findings. The patient was finally diagnosed with paraganglioma in vertebral canal superimposed on dermatomyositis.

Objective To investigate the effects of HINT1 on the growth of and apoptosis in malignant melanoma in nude mouse models established by subcutaneous xenotransplantation of human melanoma A375 cells. Methods Three groups of nude mice were subcutaneously inoculated with A375 cells transfected with pcDNA 3.1/myc-His (-) A-HINT1 (HINT1-A375), A375 cells transfected with pcDNA 3.1/myc-His (-) A empty vector (neo-A375), and untransfected A375 cells, respectively. Then, the growth of transplanted tumors was observed and tumor formation rate was calculated. Thirty-three days after the inoculation, mice were killed and tumor tissue was obtained followed by the examination of tumor weight and volume. Histopathology was performed to observe the morphological features of tumor cells and in situ end labeling technique (TUNEL)was carried out to assess the apoptosis in transplanted tumor cells. Results The tumor formation rate was consistently 100% in the three groups. The transplanted tumor in HINT1 group grew more slowly than that in the other two groups, and significant difference was observed as early as day 18 (P ＜ 0.01 ). Lower tumor weight and volume were noted in the HINT1 group compared with the neo group and A375 cell group (0.04 ±0.00 g vs. 0.23 ± 0.00 g and 0.29 ± 0.03 g, 0.06 ± 0.04 cm3 vs. 0.34 ± 0.15 cm3 and 0.43 ± 0.19 cm3 respectively, all P ＜ 0.01 ). Histopathology revealed smaller tumor nests, slight atypia of tumor cells with no obvious pathologic mitoses or necrosis in HINT1 group in comparison with the other two groups. Immunohistochemistry and TUNEL revealed that the percentage of apoptotic cells in HINT1 group was statistically higher than that in the neo group and A375 cell group (12.87% ± 1.18% vs. 3.22% ± 0.49% and 3.00% ± 0.53%, both P ＜0.01 ). Conclusion High expression of HINTl could inhibit the growth of and promote the apoptosis in malignant melanoma in nude mice subcutaneous xenotransplantation models, suggesting that HINT1 gene might be responsible for tumor suppression in human malignant melanoma.

Objective To express and purify the epitope peptide of human melanin-concentrating hormone receptor 1, and to evaluate its performance in the detection of autoantibodies in vitiligo patients. Methods The target gene encoding the epitope peptide of human melanin-concentrating hormone receptor 1 was synthesized, cloned to prokaryotic expression vector pGEX-4T-2 which was then transferred to E. coli BL21. The protein expression was induced by isopropy-β-D-thiogalactoside (IPTG) and identified with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Blocking ELISA was carried out with membrane proteins extracted from melanocytes as the blocking antigen. The antigenicity of the peptide was detected in sera from 100 patients with progressive vitiligo and 30 healthy human controls. Results The recombinant expression vector was successfully constructed, and the target protein was successfully expressed in E.coli, which was evidenced by SDS-PAGE and Western blot. With the glutathione S-transferase (GST) purification kit, the purity of the recombinant protein reached 100% when the sampling weight was less than 0.625 μg.The binding of the target protein with serum IgG antibodies from vitiligo patients could be blocked by natural membrane antigen of melanocytes. Of the 100 sera from patients with progressive vitiligo, 36 were reactive with the target protein. Conclusions The epitope peptide of human melanin-concentrating hormone receptor 1 has been successfully expressed and purified. The purified protein can bind with serum IgG antibodies from vitiligo patients, and may be applied to the detection of autoantibodies against human melanin-concentrating hormone receptor 1.

Objective To investigate the clinical and histological characteristics of trichilemmal carcinoma (TLC). Methods A clinicopathological analysis of 13 cases of TLC was carried out. Results There were 9 males and 4 females among the 13 patients with TLC who were aged from 34 to 87 years (mean: 70 years). Clinically, the tumor presented as an exophytic mass; histologically, it was characterized by the proliferation of epithelial cells and keratinization of outer root sheath. Cytologically atypical clear cells predominated in the tumor tissue. Microscopy revealed different growth patterns of tumor cells, which included solid growth pattern, tobular pattern and trabecular pattern. Periodic acid Schiff (PAS) stain demonstrated clear cells in all the tumor tissues from the 13 patients. Immunohistochemistry was performed in tissue samples from 6 patients, and showed that these samples were positive to high molecular weight cytokeratin (CK-HMW) and epithelial membrane antigen (EMA), but negative to carcinomebrynic antigen (CEA), S-100, cytokeratin 8 (CK8)and epithelial antigen(Ber-Ep4). Follow-up over 4 months to 5 years revealed neither recurrence nor metastasis in 9 cases.Conclusions TLC is a low-grade malignancy of skin adnexal tumor without distinctive clinical features, and should be differentiated from other malignant clear cell tumors of the skin.

Objective To study the effects of bFGF on the proliferation of cultured human melanocytes, and to seek a quick method for in vitro culture of human melanocytes. Methods Melanocytes were isolated from human foreskin, and divided into two parts to be cultured with or without the presence of bFGF (0.3 μg/L). Second-passage melanocytes were identified with immunochemical stain. The growth of melanocytes was observed every 3 days for 12 days. Third-passage melanocytes were treated with various concentrations (0.3 - 2.1 μg/L) of bFGF for 72 hours followed by the detection of proliferation of and trosinase activity in melanocytes. Results Human melanocytes were obtained from primary culture in medium containing certain concentrations of bFGF, which were identified with immunohistochemical stain. The morphology of cultured melanocytes varied with growth stage of cells. The bFGF-treated melanocytes appeared to grow more rapidly than untreated melanocytes. Further more, a significant increase was observed in the proliferation rate of melanocytes treated with bFGF of 0.3 and 0.6 μg/L (P＜0.05 or 0.01 ) and tyrosinase activity in melanocytes treated with bFGF of 1.5 and 1.8 μg/L (P ＜ 0.05 or 0.01 ) in comparison with the untreated melanocytes.Conclusions The addition of certain concentrations of bFGF to defined medium can benefit the primary culture of melanocytes and make it possible to get large quantities of purified melanocytes with high viability in short periods. Certain concentrations of bFGF can up-regulate the proliferation of and tyrosinase activity in melanocytes.

Objective To investigate the efficacy of autologous peripheral hematopoietic stem cell transplantation in the treatment of adult dermatomyositis. Methods A 21-year-old patient with dermatomyositis received autologous peripheral hematopoietic stem cell transplantation and was followed up for 6 years. Autologous peripheral hematopoietic stem cells were mobilized by recombinant human granulocyte colony stimulating factor (rhG-CSF) before the transplantation, and the conditioning regimens consisted of cyclophosphamide,methylprednisolone and cyclosporin. Rabbit anti-human T lymphocyte immunoglobulin began to be applied on day 3 after retransfer of stem cells. The improvement in symptoms, physical signs and biochemical indicators was observed, and hematopoietic restructuration and immunity resurrection were evaluated after the transplantation. Results After the transplantation, skin eruption greatly improved and gradually subsided. The muscle force of extremities restored from level Ⅳ before transplantation to level Ⅴ. The level of creatine kinase declined sharply after transplantation, but gradually returned to previous level. Leucocyte count began to decrease on the day of retransfer, and returned to the normal level on day 8. Immune function remained normal before and after the transplantation. Conclusion Autologous peripheral hematopoietic stem cell transplantation is an alternative treatment for severe and refractory dermatomyositis.

Objective To compare the clinical efficacy and safety of isotretinoin erythromycin gel, a gel containing isotretinoin (0.05%) and erythromycin (2%), versus adapalene gel in the treatment of mild to moderate acne vulgaris. Methods A multicenter, randomized, open, parallel-controlled clinical study was conducted. A total of 192 patients with mild to moderate (Grade Ⅰ -Ⅲ ) acne vulgaris were enrolled in this study according to the grading criteria for acne severity in guidelines for the treatment of acne in China. Efficacy analysis was carried out in 169 patients and safety analysis in 190 patients. The patients were classified into trial group (n = 86) and control group (n = 83 ) to be treated with isotretinoin erythromycin gel or adapalene gel once a night for 6 weeks. Patients were evaluated at the baseline, on week 2, 4 and 6 during the treatment for the count of comedones (both open and closed), inflammatory papules and pustules, severity of acne and local or general adverse effects. Results After the start of treatment, the response rate gradually increased and severity of acne decreased in both groups. On week 6, the total response rate was 51.16% in the trial group and 40.96% in the control group (P ＞ 0.05), while a greater reduction in the count of pustules and inflammatory lesions was observed on week 4 and 6 in the trial group with a lower severity grade of acne compared with the control group (P ＜ 0.05 or 0.01 ). Adverse reactions were similar in both groups and manifested as tolerable local irritation. Conclusions The efficacy of isotretinoin erythromycin gel is similar to that of adapalene gel in the treatment of mild to moderate acne vulgaris, however, isotretinoin erythromycin gel seems superior to adapalene gel in reducing inflammatory lesions and rapidly improving severity of acne vulgaris.