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Expression of insulin-like growth factor-Ⅱ (IGF-Ⅱ) mRNA-binding protein-3 in malignant melanoma and benign nevi

Chongmei XIN ; Xiulian XU ; Amei LI ; Yiqun JIANG ; Hao CHEN ; Xuesi ZENG ; Jianfang SUN

Chinese Journal of Dermatology.2010;43(7):467-470. doi:10.3760/cma.j.issn.0412-4030.2010.07.008

Objective To investigate the expression of insulin-like growth factor-Ⅱ(IGF-Ⅱ) mRNAbinding protein-3 (IMP3) in tissue of benign nevi and malignant melanoma,and to evaluate the role of IMP3 in the development and diagnosis of malignant melanoma.Methods Immunohistochemistry was performed to measure the expression of IMP3 in tissue samples from 28 cases of malignant melanoma,8 Spitz nevi,6 dysplastic nevi and 25 benign nevi.Results Immunohistochemically,IMP3 was observed in 23 of 28 melanoma samples,4 of 8 Spitz nevus samples and 2 of 6 dysplastic nevus samples,but not in benign nevus samples.The expression level of IMP3 wag significantly higher in tissue of melanoma than in that of Spitz nevi and dysplastic nevi (both P<0.05),also higher in tissue of aggressive melanoma than that of melanoma in situ(P<0.01).Conclusions IMP3 seems to be a biomarker for the progression of benign nevi to malignant melanoma,and may be utilized to distinguish melanoma from benign nevi.

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Association between estrogen receptor gene polymorphism and systemic lupus erythematosus in women of Han nationality from Jiangxi province

Ougen LIU ; Qingshui HUANG ; Zhibing DUAN ; Tianlun HUANG ; Zhigang LIU

Chinese Journal of Dermatology.2010;43(7):460-462. doi:10.3760/cma.j.issn.0412-4030.2010.07.005

Objective To explore the association between estrogen receptor gene polymorphism and systemic lupus erythematosns in women of Han nationality from Jiangxi province.Methods Whole blood samples were obtained from 95 female patients with SLE of Han nationality (18 with pre-menarche onset and 77 with post-menarche onset) and 100 healthy female controls from Jiangxi province.Genomic DNA was extracted from these samples followed by the analysis of polymorphism at Pvu Ⅱ and Xba I sites of estrogen receptor alpha gene with PCR-restriction fragment length polymorphism (RFLP).The frequencies of alleles and genotypes were compared between the control and patients and between patients with pre-menarche onset and those with post-menarche onset.Results The frequency of P allele was higher in patients with SLE than in the controls (35.8% vs 27%,P<0.05),but lower in patients with pre-menarche onset than in those with post-menarche onset (19.4% vs 39.6%,P<0.05).Further more,an increment was observed in the frequency of Xxpp genotype in patients with pre-menarche onset compared with normal controls and patients with postmenarche onset (16.7% vs 1% and 1.3%,both P<0.05).Conclusions The frequency of P allele in ER alpha gene is increased in female patients with SLE of Han nationality from Jiangxi province.The Xxpp genotype of ER gene may be associated with the susceptibility to pre-menarche onset SLE.

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Transplantation of individualized cultured autologous melanocytes for the treatment of vitiligo

Dimin ZHANG ; Weisong HONG ; Lifang FU ; Guopei QIAN ; Aie XU

Chinese Journal of Dermatology.2010;43(10):721-725. doi:10.3760/cma.j.issn.0412-4030.2010.10.015

Objective To evaluate the therapeutic effect of transplantation of autologous melanocytes cultured with individualized medium in vitiligo. Methods Donor skin was obtained by suction blisters from a normally pigmented area of the abdomen of 155 patients with vitiligo. The roof of the blisters was clipped and digested with trypsin, then the suspension of epidermal cells and melanocytes were cultured in Hu16 medium.The cell division time (DOT) and melanin content of cultured melanocytes were measured followed by the adjustment of concentration of fetal calf serum, cytokines and cAMP elevating agents based on the DOT,melanin content and morphology of melanocytes for the individualized culture of melanocytes. After 2 - 5 passages, melanocytes were harvested and inoculated into ultrapluse CO2 laser-denuded lesions. All patients were followed up for at least 6 months. Results One hundred and fifty-five vitiligo patients with 204 lesions were treated with transplantation of autologous melanocytes. Of the 155 patients, 119 received 1 session of transplantation, 36 received 2 to 4 session of transplantation. Cells were expanded by 50 - 80 times in vitro after individualized culture. Repigmentation was more than 50% in 84.8% of these lesions, more than 90% in 52.94% of the lesions. A homogeneous skin color was obtained in repigmented skin, and no scarring or other side effects were observed. No influence was noted on the outcome of transplantation for sex, age, course of disease or lesion size of patients. Segmental vitiligo showed better response than vitiligo vulgaris: the effective rate and cure rate were 93.62% and 65.96% respectively for segmental vitiligo, 82.16% and 49.04% respectively for vitiligo vulgaris. Lesions located on the arms and legs (not including elbows and knees) showed the best response, with a cure rate of 73.08%, whereas acral sites were the most difficult area to repigment, with a cure rate of just 25.93%. Conclusions Individualized culture can significantly increase the success rate of melanocyte culture and expanding times of melanocytes. Transplantation of cultured autologous melanocytes is an effective modality deserving clinical application in the treatment of stable vitiligo, with the advantage of treating large depigmented area with melanocytes from a small donor site.

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Effects of intense pulsed light irradiation on the content of collagen fibers, elastic fibers and hyaluronic acid in Kunming mouse skin

Liehua DENG ; Saijun LIU ; Yunfeng HU ; Gang ZHAO ; Ze LIN ; Yongkeng ZHAO

Chinese Journal of Dermatology.2010;43(10):705-708. doi:10.3760/cma.j.issn.0412-4030.2010.10.011

Objective To investigate the effects of intense pulsed light (IPL) irradiation on the content of collagen fibers, elastic fibers and hyaluronic acid in Kunming mouse skin. Methods The dorsal skin of mice was divided into two areas: the right area was irradiated with IPL, and the left remaining unirradiated served as the control. Skin specimens were taken from the back of mice on day 1, 3, week 1, 2, 4 and 8 after the irradiation and subjected to staining with HE, sirius red and Gomori aldehyde-fuchsin for examinations of histological changes, type Ⅰ and Ⅲ collagen fibers and elastic fibers. The hydroxyproline and hyaluronic acid content in skin tissues of mice was determined with ultraviolet spectrophotometry and radioimmunoassay respectively. Results After irradiation, a significant increase was observed in dermal thickness on week 2 (t =4.623, P＜ 0.05), 4, and 8 (t = 3.904, P＜ 0.05), in type Ⅲ collagen fiber (t = 5.129, P＜ 0.05) on week 1,in type Ⅰ and Ⅲ collagen fibers on week 2, 4 and 8 (both P＜ 0.05), in elastic fibers from week 2 to 8 (P ＜0.05), and in hydroxyproline content from week 1 to 8 (all P ＜ 0.05) in the skin of mice compared with unirradiated mice. In detail, dermal thickness increased by 18.71% on week 4, and type Ⅲ collagen fiber by 40.54% in irradiated mice compared with unirradiated mice. Further more, the hyaluronic acid content was elevated from day 1 to 3, but gradually declined from week 1 to 8, and remained statistically higher from day 1 to week 8 (P ＜ 0.05) in irradiated mice compared to unirradiated mice. Conclusion IPL irradiation could induce an increase in the content of collagen fiber, elastic fiber and hyaluronic acid in the dorsal skin of mice.

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Construction of models for persistent genital tract infection with Chlamydia trachomatis in mice with low serum iron

Yong LIU ; Yuanjun LIU ; Weifeng YAO ; Caihong SHENG ; Hui ZHU ; Yan LI ; Quanzhong LIU

Chinese Journal of Dermatology.2010;43(10):702-704. doi:10.3760/cma.j.issn.0412-4030.2010.10.010

Objective To construct a model for persistent genital tract infection with Chlamydia trachomatis in mice with low serum iron. Methods Iron deficiency was induced in female BALB/c mice by lowiron diet. Mice with or without iron-deficency were inoculated intravaginally with direct instillation of 50 μl of bacterial suspension containing 3.4344 × 107 IFU/ml of C. trachomatis serovar E elementary body 1 week after intramuscular injection of progesterone. Moxifloxacin and ferrous sulfate, alone or in combination were given to inoculated mice with iron deficiency daily for 1 week. Subsequently, vaginal discharge was obtained from mice and observed under an immunofluorescence microscope, and tissue specimens were resected from the uterus,uterine tube and ovary of mice and subjected to pathological examination at various time points. Results Compared with normal-iron mice, iron-deficient mice were infected with C. trachomatis for a longer duration (more than 18 weeks). After 18-week infection with C. trachomatis, there was an obvious chronic inflammation and lymphocytic infiltration in tissues specimens from the uterus,uterine tube and ovary of mice. Immunofluorescent examination of vaginal discharge for C. trachomatis turned negative in 60% of iron-deficient mice treated with both moxifloxacin and ferrous sulfate, but not in any of those receiving moxifloxacin or ferrous sulfate alone. Conclusion As animal models show, low serum iron may contribute to the persistent genital tract infection with C. trachomatis.

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Effects of irradiation with 1064-nm Q-switched Nd:YAG laser on melanogenesis in cultured PIG cells in vitro

Ling WANG ; Chengxin LI ; Dongning ZHU ; Zhe JIAN

Chinese Journal of Dermatology.2010;43(10):713-716. doi:10.3760/cma.j.issn.0412-4030.2010.10.013

Objective To study to the effect of 1064-nm Q-switched Nd:YAG laser irradiation on the melanogenesis in a human epidermal melanocyte line PIG. Methods Cultured PIG cells were irradiated with 1064-nm Q-switched Nd:YAG laser (Medlite C6) at different energy densities for 10 times. After additional culture for various durations, cell viability was detected by MT assay, tyrosinase activity by dopa oxidation assay, mRNA and protein expressions of tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 by real-time quantitative fluorescent RT-PCR and Westen blotting respectively, Results The irradiation with Q-switched Nd:YAG laser at energy densities from 1 to 3 J/cm2 had no obvious effect on the viability of PIG cells. After irradiation with Nd:YAG laser at 1 J/cm2, PIG cells showed a significant increase in the tyrosinase activity,mRNA expressions of tyrosinase and TRP-1 compared with unirradiated cells (0.563 ± 0.014 vs 0.501 ±0.019, 1.40±0.11 vs 1.0, 1.28 ± 0.03 vs 1.0, all P＜ 0.05), but both the mRNA (0.91 ± 0.17 vs 1.0, P＞0.05) and protein expressions of TRP-2 experienced no significant changes before and after the irradiation.However, a significant decrease was noted in PIG cells irradiated with Nd:YAG laser at 3 J/cm2 in tyrosinase activity, mRNA and protein expressions of tyrosinase (0.70 ± 0.02 vs 1.0, 0.64 ± 0.05 vs 1.0, both P ＜ 0.05),TRP-1 (0.73±0.04 vs l.0, 0.86±0.17 vs l.0, both P＜0.05) andTRP-2 (0.68±0.04 vs l.0,0.69±0.11vs 1.0, both P ＜0.05) in comparison with unirradiated PIG cells. Conclusions The 1064-nm Q-switched Nd:YAG laser irradiation may affect the melanogenesis in PIG cells. With no influence on cell viability, the 1064-nm Q-switched Nd:YAG laser at 1 J/cm2 could enhance melanogenesis, while that at 3 J/cm2 could suppress melanogenesis, in PIG cells.

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A preliminary study on the expression and function of μ-opioid system in the epidermis of patients with atopic dermatitis

Jie CHEN ; Mei SHI ; Yifeng GUO ; Zhirong YAO

Chinese Journal of Dermatology.2010;43(10):709-712. doi:10.3760/cma.j.issn.0412-4030.2010.10.012

Objective To investigate the expression of μ-opioid system in the epidermis of patients with atopic dermatitis and its role in the pathogenesis of atopic dermatitis. Methods Thirty-two mice were equally divided into 4 groups, negative control group, pre-treatment group, naloxone group, and physiological saline group. Ovalbumin was used to sensitize mice in pretreatment group, naloxone group, and physiological saline group for 7 weeks, then, mice in naloxone group and physiological saline group were treated with intracutaneous naloxone or physiological saline solution for 1 week, respectively. Mice were killed in negative control group and pre-treatment group at the end of sensitization, and in naloxone group and physiological saline group after 1-week injection with naloxone or physiological saline, skin tissues were obtained from the back of killed mice and subjected to histological examination with HE staining and quantitative fluorescent PCR for the detection of mRNA expression of μ-opioid receptor (MOR) and its ligand (β-endorphin) in epidermis. The atopic dermatitis severity index of lesions and histological changes were assessed before and after the treatment. Results In comparison with the negative control mice, the epidermal expression level of MOR was signifieantly decreased (t = 2.549, P ＜ 0.05 ) in pre-treatment group, but increased in naloxone group and showed no statistical difference from the negative control group (t = 0.671, P ＞ 0.05). No significant difference was observed in the epidermal β-endorphin mRNA expression between negative control group and pre-treatment group or naloxone group (both P ＞ 0.05 ). The improvement of lesions could be visualized after treatment with naloxone (t = 8.338, P ＜ 0.01 ), which was concordant with the histological changes in naloxone group. Conchusions As an antagonist of MOR, naloxone can restore the expression of epidermal MOR in mice model for atopic dermatitis, and shows a certain efficacy in the treatment of atopic dermatitis, which proves that μ-opioid system is somewhat associated with the pathogenesis of atopic dermatitis.

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Changes in the number and biological ability of endothelial progenitor cells from peripheral blood of patients with systemic lupus erythematosus (SLE)

Peimei ZHOU ; Yonghong LU ; Xia XIONG ; Xiaoyun CHENG ; Yanjun ZHU ; Shihua ZENG ; Tao CHEN ; Qingbiao WA ; Qi CAI ; Qianming CHEN ; Cunhuo JIANG

Chinese Journal of Dermatology.2010;43(10):698-701. doi:10.3760/cma.j.issn.0412-4030.2010.10.009

Objective To analyze the changes in number and biological ability of endothelial progenitor cells (EPCs) from peripheral blood of SLE patients. Methods Mononuclear cells (MNCs) were isolated by Ficoll density gradient centrifugation from peripheral blood of 20 female SLE patients and 20 healthy female controls. EPCs were identified by double staining using antibodies to CD34 and CD133, or antibodies to CD133 and vascular endothelial growth factor receptor 2 (VEGFR2). Phycoerythrin (PE) conjugated antiCD34, fluorescein isothiocyanate (FITC) conjugated anti-CD133 and APC conjugated anti-VEGFR2 antibodies were used in a three color flow cytometric analysis to determine the percentage of EPCs in peripheral MNCs.The proliferation and migration ability of EPCs were measured by MTT assay and modified millicell chamber assay, respectively. The adhesion activity of EPCs was evaluated by counting the number of adherent cells.Results The percentage and proliferation rate of EPCs in peripheral MNCs from female SLE patients were significantly lower than those from the healthy controls(4.49% ± 1.66% vs 20.81% ± 4.14%, 23.11% ± 3.16%vs 35.65% ± 1.74%, both P ＜ 0.01 ). The migration and adhesion ability of EPCs from SLE patients was impaired compared with those from the healthy controls (12.00 ± 2.12 vs 23.60 ± 3.0 cells/field, 22.43 ± 4.43vs 36.43 ± 3.69 cells/filed, both P ＜ 0.01 ). Conclusion There is a decrease in the number and an impairment in biological ability of EPCs in SLE patients.

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Ultrastructure of skin lesions and mutations in the FERMT1 gene in a patient with Kindler syndrome

Zhimiao LIN ; Yanhong TAN ; Zhihong MA ; Quan CHEN ; Yun WANG ; Xiuying TANG ; Suxia WANG ; Yong YANG

Chinese Journal of Dermatology.2010;43(10):677-679. doi:10.3760/cma.j.issn.0412-4030.2010.10.002

Objective To study cutaneous ultrastructural changes and FERMT1 gene mutations in a patient with Kindler syndrome. Methods Clinical data were collected, and tissue samples obtained from the lesions of poikiloderma were observed by using transmission electron microscopy. Fifteen coding exons and their flanking sequences of the FERMT1 gene were amplified by PCR and DNA sequencing was followed.Results Reduplication of lamina densa was seen between the dermal-epidermal junctions of the lesional skin. The patient was found to be homozygous for a novel splice-site mutation (IVS9 + 1G ＞ A) in FERMT1 gene, and his parents were heterozygous for it. The mutation was undetected in fifty normal control individuals.Conclusions Transmission electron microscopy may serve as an ancillary examination for the diagnosis of Kindler syndrome. The IVS9+1G＞A mutation of FERMT1 gene may contribute to the clinical phenotype of Kindler syndrome in this patient.

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Changes of molecular markers in cultured skin stem cells exposed to ultraviolet B (UVB) in vitro

Jing WANG ; Bo CHENG

Chinese Journal of Dermatology.2010;43(10):726-729. doi:10.3760/cma.j.issn.0412-4030.2010.10.017

Objective To investigate the changes of molecular markers in cultured skin stem cells exposed to UVB in vitro. Methods Skin stem cells were isolated and cultured according to their adherasion ability,and identified by immunohistochemistry using anti-K15 and anti-β-integrin antibodies. Then, a part of the skin stem cells were irradiated with UVB at 10 mJ/cm2 for 2 times. After 24-hour additional culture, the expressions of CD34, beta-catenin and p53 were detected with immunohistochemistry. Results Skin stem cells showed a high density in culture free of irradiation, which were round or polygon with a clear shape, well-distributed cytoplasm, high N/C ratio; mitotic cells could be seen. In unirradiated skin stem cells, beta-catenin was expressed predominantly in cell membrane and cytoplasm, with a positive expression rate of 64.74% and 8.4%in membrane and cytoplasm respectively; p53 was expressed mainly in cell cytoplasm and nuclei, with a positive expression rate of 6.9% in cell nuclei. After exposure to UVB, skin stem cells decreased in cell density and N/C ratios with a deformed and anomalous shape, vacuoles were present in cytoplasm, and some cells experienced karyopyknosis or apoptosis. Additionally, in irradiated cells, beta-catenin was expressed predominantly in cytoplasm with a positive expression rate of 64.74% and 0 in cytoplasm and nuclei, respectively; p53 was expressed mainly in nuclei with a positive expression rate of 100%. CD34 was detected in neither unirradiated nor irradiated skin stem cells. Conclusion UVB can promote beta-eatenin to accumulate in cytoplasm as well as beta-catenin and p53 to migrate from cytoplasm to nuclei.

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