Photoprotective Effects of Hydroxychloroqine and TCMs on Human Keratinocytes Damaged from Ultraviolet Irradiation

Objective: To investigate damage effects of ultraviolet irradiation on eternal keratinocyte-HaCaT cells and to evaluate photo-protective efficiency of hydroxychloroqine and Traditional Chinese Medicines(epigallocatechingallate[EGCG], baikal skullcap root and szechwan lovge rhizome) on HaCaT cells damaged by middle wave ultraviolet(UVB) irradiation. Methods: Subconfluent HaCaT cells were sham or UVB irradiated and treated with above TCM agents. The damage degree of HaCaT cells was observed by a light microscop. Cell growth was recorded by cell count and cellular activity was detected by MTT method. The secretion amount of IL-6 and TNF-α was measured by ELISA. Results: The irradiation damage of HaCaT cells was depended on the irradiated dosages and cellular activity was reduced by 36%-80%, with a maximum decrease over 90% after 72 h. The intervention of the above drugs may increase the cellular activity by 10%-72%. The photo-protective efficiency was more apparent in EGCG (from 1.19±0.07 to 1.28±0.06, P<0.01) than that in hydroxychloroqine (from 0.43±0.04 to 0.96±0.04, P<0.05). The other two tested drugs also showed photo-protective effect(from 0.44±0.07 to 1.21±0.02, P<0.05). As to cytokine secretion, EGCG could decline the secretion amount of IL-6 and TNF-α apparently. Hydroxychloroqine and baikal skullcap root baikal skullcap root could only reduce the secretion of IL-6. The secretion of IL-6 and TNF-α could not be inhibited by szechwan lovge rhizome. Conclusion: The injury effect of UVB irradiation on cultured keratinocytes is dose-dependent and the tested drugs have photo-protective potency. Inhibition of cytokine secretion may be one of the mechanisms related to reducing the damage effect of UVB irradiation.

CONSTRUCTION AND IMMUNOGENICITY OF HUMAN PAPILLOMAVIRUS TYPE 6B L1 RECOMBINANT PLASMID

To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immu-nogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into an eukaryotic ex-pression plasmid (pcDNA3.1). The recombinant plasmid was transfected into COS-7 cells. Western blot were performed to detect whether L1 protein can be expressed in eukaryotic cells. Eighteen female BALB/c mice were tested for immunoge-nicity study. Results The recombinant plasmid (pcDNA3.1-HPV6bL1) was verified as HPV6b L1 gene by sequencing. Western blot showed specific strip. Anti-L1 protein antibodies could be detected in the mice’s sera inoculated with pcDNA3.1-HPV6bL1. Similarly, IL-4, IL-2, and IFN-γ were increased in the same mice. Conclusion HPV6b L1 recombinant plasmid was constructed successfully which had immunogenicity for BALB/c mice. It provided experimental evidence for the research of DNA vaccine of condyloma acuminata..-

Construction of Eukaryotic Expression Plasmid pcDNA3-FHIT and Its Transient Expression in COS-1 Cells

Objective: To construct eukaryotic expression plasmid pcDNA3-fragile histidine triad(FHIT) and obtain its transient expression in COS-1 cells. Methods: FHIT gene was cloned from normal human thyroid tissue by RT-PCR and then inserted into eukaryotic expression vector pcDNA3. After the sequence was confirmed, the recombinant plasmid pcDNA3-FHIT was transfected into COS-1 cells by cation liposome. The transient expression in the cells was measured by immunocytochemistry. Results: The sequence of FHIT in pcDNA3 was correct and high expression was obtained in COS-1 cells. Conclusion: The eukaryotic expression plasmid pcDNA3-FHIT was constructed successfully and could highly express FHIT protein in COS-1 cells. This will be potentially useful for the research on gene therapy.

Analysis of the T-Lymphocytes and Their Subpopu-lations of the Peripheral Blood in Patients with Urticaria

Objective: To study the function of cellular immunity of patients with urticaria. Methods:T-lymphocytes subpopulations of the peripheral Mood ire 60 patients with urticaria and 40 healthy controls were examined by flaw cytometry. Results: The number of CD3+ and CD4+ cells in the urticaria, group were significantly lower than tliose in the control group (P<0. 01) , especially in patients with acute urticaria. Conclusion: There was immunologic dysfunction of T lymphocyte in the patients with urticaria, and not only humoral immunity takes part but also cellular immunity plays a certain role in the pathogenesis of urticaria.

A summary about dendritic cells in skin diseases

Dendritic cellls (DCs) comprise an es se ntial component of the immune system, are crucial in the initiation of antigen s pecific immune responses. In this summary we focus on summarizing on the central role of DCs in skin diseases: Bullous dermatoses, Dermatitis, Psoriasis, Lichen Planus , Graft-versus-host disease, Connect Tissue Diseases, Virus Diseases, Fungi Diseases, HIV, Urticaria, Urticaria pigmentosa, Mastocytosis, Tumour, Sola r dermatoses. Moreover,in this summary we review the distribution and phenotype of DCs in human skin. Markers and phenotyps 's study have provided strong suppo rt for a concept in which DCs play an important role in the pothogenesis of som e skin diseases.

Combination of Circumcision and Microwave in Treatment of Genital Warts in Uncircumcised Patients

Objective: To investigate the efficacy of combina-tion of circumcision and microwave on genital warts in uncircumcised men.Methods: A randomized, prospective study of 109 uncircumcised adult men with genital warts was con-ducted in a STD clinic in Zhanjiang, Guangdong. One group (n=54) received microwave therapy only, while the other group (n=55) was taken the combination of circumcision and microwave therapy. The recurrences were observed at the end of months 3, 6 and 12, and operative complications were also recorded.Results: There were no significant differences in the mean age and duration of the disease between two groups (P>0.05). No serious operative complications were documented. The recurrence rate in circumci-sion plus microwave group was markedly lower than that in microwave group (12.7% vs 29.6%, P<0.05),while the differences in early and late recurrences between two groups showed no statistical significance(P>O.05).Conclusion: Circumcision can be safely performed under local anesthesia in an outpatient setting. Com-bination of circumcision and microwave can produce excellent effect as well as less tissue damage,therefore, it may be ideal for uncircumcised patients with extensive condvlomas.

兰索拉唑引起的致死性中毒性表皮坏死症

Toxic epidermal necrolysis (TEN) is a serious cutaneous reaction and is most commonly drug induced. It is associated with significant morbidity and mortality. We describe a patient who developed fatal TEN after re-exposure to lansoprazole. Three years previously he presented with erythema multiforme due to the same drug. To our knowledge this is the first published report of TEN occurring with lansoprazole.

富含镁的死海盐溶液可提高皮肤屏障功能、水合程度及减轻异位性干燥皮肤的炎症

Magnesium salts, the prevalent minerals in Dead Sea water, are known to exhibit favorable effects in inflammatory diseases. We examined the efficacy of bathing atopic subjects in a salt rich in magnesium chloride from deep layers of the Dead Sea (Mavena. Dermaline Mg46 Dead Sea salt, Mavena AG, Belp, Switzerland). Volunteers with atopic dry skin submerged one forearm for 15 min in a bath solution containing 5% Dead Sea salt. The second arm was submerged in tap water as control. Before the study and at weeks 1- 6, transepidermal water loss (TEWL), skin hydration, skin roughness, and skin redness were determined. We found one subgroup with a normal and one subgroup with an elevated TEWL before the study. Bathing in the Dead Sea salt solution significantly improved skin barrier function compared with the tap water-treated control forearm in the subgroup with elevated basal TEWL. Skin hydration was enhanced on the forearm treated with the Dead Sea salt in each group, which means the treatment moisturized the skin. Skin roughness and redness of the skin as a marker for inflammation were significantly reduced after bathing in the salt solution. This demonstrates that bathing in the salt solution was well tolerated, improved skin barrier function, enhanced stratum corneum hydration, and reduced skin roughness and inflammation. We suggest that the favorable effects of bathing in the Dead Sea salt solution are most likely related to the high magnesium content. Magnesium salts are known to bind water,influence epidermal proliferation and differentiation,and enhance permeability barrier repair.

Study on the Expression of IL-18, AcPL and IL-18BP mRNA in the Psoriatic Lesions

To investigate the role of IL-18 and IL-18 binding protein (IL-18BP) in the pathogenesis of psoriasis, PT-PCR was used to semi-quantitatively analyze the mRNA expression of IL-18, AcPL (accessory protein-like) and IL-18BP in 12 psoriatic lesions and in 6 normal skin tissues. The results showed that IL-18 and AcPL mRNA were present in all specimens from 6 normal skin and 12 pieces of psoriatic skin, and IL-18 and AcPL showed relatively stronger intensity in the psoriatic skin compared with those seen in normal skin (P<0.01). Amplified bands indicating the expression of IL-18BP mRNA were weakly positive in all samples of normal skin. In contrast, the PCR products for IL-18BP were distinctly visible in all specimens of psoriatic lesions (P<0.01). These findings demonstrate that IL-18 is constitutively synthesized by human keratinocytes and involved in the development of the Th1 response in psoriatic lesions, and its bioactivity appears to be closely regulated by cutaneous inflammation.

Relationships of tumor inflammatory infiltration and necrosis with microsatellite instability in colorectal cancers

AIM:The relationships between microsatellite instability (MSI) and survival in colorectal cancer patients are not consistent. The favorable survival of patient with MSI has been suggested to be related to pronounced inflammatory infiltration; however,the reason for non-association of MSI with survival is unclear. Our aims were to investigate the associations of inflammatory infiltration and tumor necrosis (TIM) with microsatellite status and clinicopathological factors in colorectal cancer patients in whom MSI was not related to survival. METHODS: Three hundred and one colorectal adenocar-cinomas were evaluated for inflammatory infiltration and 300 for TN under light microscope. RESULTS: Low infiltration at invasive margin (X2=3.94, P= 0.047) and in whole tumor stroma (X2=3.89, P= 0.049) was associated with MSI, but TN was not (X2=0.10, P = 0.75). Low infiltration was related to advanced stage (X2 = 8.67, P= 0.03), poorer differentiation (X2=8.84, P= 0.03), DNA non-diploid (X2=10.04, P= 0.002), higher S-phase fraction (X2=11.30,P=0.004),positive p53 expression (X2=7.94,P=0.01),and worse survival (P = 0.03 for both univariate and multivariate analyses). Abundant TN was related to advanced stage (X2=17.74, P= 0.001) and worse survival (P= 0.02 for univariate, and P= 0.05 for multivariate analysis). CONCLUSION: The result that high inflammatory infiltration was not related to MSI might help explain the non-association of MSI with survival in colorectal cancer patients.

Treatment of folliculitis keloidalis nuchae with He-Ne Laser

AIM:To verify curative effect of He Ne laser on folliculitis keloidalis nuchae.METHODS:103 cases with folliculitis keloidalis nuchae were divided into the treatment group(73 cases) and the control group(30 cases). Local application and oral antiauxins was used in the two groups,He Ne laser radiation was used in the treatment group and then curative effect 15 days later were compared.RESULTS:The cure rats in the treatment group and control group were respectively 71.2%(52/73) and 30%(9/30)(P< 0.01) while curative days were respectively 9.2 days and 11.9 days (P< 0.01).CONCLUSION:In treatment of folliculitis Keloidalis nuchae,the application of He Ne Laser radiation can effectively reduce the course,pain and side effect of treatment,and raise the curative effect.

Detection of Anticardiolipin Antibody in Systemic Lupus Erythematosus and Its Clinical Significance

Objective To determine the frequency and the clinical significance of anticardiolipin antibodies (ACA) in patients with systemic lupus erythematosus( SLE).Methods The serum ACA in 30 patients with SLE and 30 sera from healthy volunteers was detected by the enzyme linked immunosorbend assay (ELISA).Results In normal group the binding index(BI) oflgG, IgM and IgA type of ACA was 1.48 ?

Investigation on etretin effects on expression of Fas/FasL ligand and apoptosis in cultured human keratinocytes

Objective:To further illuminate a possibme mechanism of Fas/FasL in the treatment of psoriasis, the expression of it and apoptosis of KC were investigated. Methods: With cell culture,immunocytochemistry, the expression of Fas/FasL protein after the treatment with etretin was observed in cultured human normal keratinocytes. Then, the state of apoptosis in cultured keratinocyte after stimulation with etretin was detected with FACS(Fluorescence Activited Cell Sortor). Results:① There was no expression of Fas/FasL protein in the cultured normal human KC. ②After the treatment with etretin, the strongest expression of FasL protein appeared after 16h,so did Fas after 40h.③ The higher the concentration of etretin,the stronger the expression of Fas/FasL protien.Under the same condition,the expression of Fas is stronger than that of FasL.④ Keratinocytes' apoptosis occurred after stimulation with etretin. Conclusion: Fas/FasL system wasn't involved in apoptosis in cultured normol human keratinocytes.But during limited period, the apoptosis of KC could be induced by (etretin),thus it can antagonize benign proliferate of keratinocytes.Our data showed up-regulation of the expression of Fas/FasL and apoptosis in cultured human keratinocytes stimulated by etretin, and its function may be involved in the therapeutic machanism of psoriasis.

Analysis of T-lymphocyte subtypes of the peripheral blood and skindelayed-type hypersensitivity in patients with hyper-IgE syndrome

Objective: To study the function of ce ll ular immunity in patients with hyper-IgE syndrome (HIE). Methods: T-lymphocyte subtypes of the peripheral blood and cutaneous delayed-typ e hypersensitivity (DTH) response to two recall antigens, tetanus toxoid (TT) an d purified protein derivative(PPD), were measured in five patients with HIE and 15 healthy controls, respectively. Results: The CD4+ cell cou nts in HIE group were significantly lower than those in control group (P<0. 01). In contrast, CD8+ cells were significantly higher than those in the contro l (P<0.01). The induration sizes of DTH response to two recall antigens wer e smaller in HIE group than those in the control group (P<0.01). Co nclusion: There was an immunologic dysfunction of T lymphocytes in the p atients with HIE.

Studies on arsenic trioxide induced mice melanoma Cloundman S91 cell apoptosis

Objective: To investigate the effects of various concentrations As 2O 3 on malignant melanoma. Methods: The viability of Cloundman melanoma S91 cells treated with As 2O 3 was measured by MTT [3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide] assay. The apoptosis was determined by transferase-mediated dUTP nick end labeling (TUNEL) detection, and the morphology of apoptotic cells was observed through transmission electron microscopy (TEM). The cells growth phase were analyzed by flow cytometry (FCM). Results: A time-and dose-dependent decrease in cell viability was induced in S91 cells after treatment with As 2O 3 at the concentration of 1-5 μmol/L respectively. The TUNEL indices were 0.033±0.018, 0.062±0.012, 0.102±0.016, 0.132±0.031, and 0.162±0.027 respectively, which were much higher compared with the control group (0.017±0.004, P<0.01). The flow cytometry showed that hypodiploid peak after treatment with 3 μmol/L and 5 μmol/L of As 2O 3 for 48 h were 9.99% and 17.59% respectively, which increased significantly compared with the control cells (3.05% P<0.01,). The apoptotic morphology observed by transmission electron microscope showed the chromatin became condensed and attached to the inner surface of nuclear membrane. Conclusion: Arsenic trioxide can induce melanoma Cloundman S91 cell apoptosis at the concentration of 1-5 μmol/L, which will provide enhanced benefit in melanoma therapy.

Intracellular Cytokine Expression Profile in Patients with Condyloma Acuminata

Objective: To study the immunological mechanisms of Condyloma acuminata(CA) through investigating T lymphocyte subset levels and cytokine profile in the peripheral blood of patients with Condyloma Acuminata.Methods: Tricolor and bicolor immunofluorescent staining antibody of cell surface antigen and intracel-lular IL-2, IL-4, IL-12, IFN- γ in CD4^+ and CD8^+ T-lymphocytes from 20 patients with CA were performed and followed by flow cytometry.Results: The number of CD3^+T, CD4^+T-lymphocytes cells and CD4^+/CD8^+T cells ratio were significantly decreased(P<0.01) in patients with CA Compared to controls, and IL-2, IL-12, IFN- γ production in CD4^+T cells was decreased(P<0.01), IL-4 and IFN- γ produc-tion in CD4^+T cells was not significantly different(P>0.05), while IL-2 and IL-12 production in CD8^+Tc cells was decreased(P<0.01),whereas IFN- γ and IL-4 pro-ducing in CD4^+T cells were of no significantly difference(P>0.05).Conclusions: There was an imbalance of T lympho-cyte subsets, Th1/Th2 cytokines and Tc1/Tc2 in the peripheral blood of CA patients, which may play an important role in the pathogenesis and progression of CA.

Fas and Bcl-2 Expression on T Lymphocyte Subsets in the Peripheral Blood of Relapsing Patients with Condyloma Acuminatum

Objective: To study the expression of Fas and Bcl-2 proteins on T lymphocyte subsets in the peripheral blood of relapsing patients with condyloma acuminatum(CA) and healthy controls.Methods: Flow cytometry (permeabization and staining procedure with conjugated antibodies) was used.Results: We observed that the expression of Fas protein on CD4^+ T lymphocyte subset of CA patients was significantly higher than that of healthy controls( P<0.01 ).Conclusions: Increased expression of Fas proteinon CD4^+ T lymphocyte subset may be a cause of de-creased percentage of CD4^+ T lymphocyte subset. This induces the increased ratio of CD4^+/CD8^+.

Effect of HSV-2 Infected Monocytes on the Production of TNF- α and IL-6

Objectiwe: In order to detect the role of monocytes in HSV-2 infection, we studied the effect of herpes sim-plex Virus-2 infection on the production of tumor ne-crosis factor (TNF- α ), interleukin-6 (IL-6) secreted by monocytes. Methods: Monocytes were infected by HSV-2 (333 Strain). Culture supernatants were collected at 1, 3,5, 7 days post-infection. The levels of TNF- α, IL-6 were measured by enzyme-linked immunosorbent as-say (ELISA). Results: The levels of TNF- α secretion by mono-cytes significantly decreased on first day post-infection. The levels of IL-6 significantly decreased on first and third days post-infection, and then gradu-ally increased to the control on seventh day post-infection.Conclusions: TNF- α and IL-6 production by mono-cytes was inhibited during HSV-2 infection. The pro-duction of cytokines may play an important role in herpes simplex viurs-2 pathogenicity and immunity.

Expression of Activated Epidermal Growth Factor Receptor and Transcription Factor E2F in Condyloma Accuminata

Objective: To study the expression of activated epi-dermal growth factor receptor (EGFR) and transcrip-tion factor E2F (E2F) in Condyloma Accuminata(CA) patients. Methods: Immunofluorescent techniques were used to investigate the expression of activated EGFR and E2F in CA patients. Results: The expression of activated EGFR on the membrane of epithelial cells in CA lesions was sig-nificantly greater compared to expression levers in the control group (P<0.01). Moreover, the co-expres-sion of activated EGFR and E2F was significantly in-creased compared to the control group (P<0.01).Conclusion: Our observations suggest that the in-crease in activated EGFR expression may stimulate hyperplasia in CA patients through the activation of transcription factor E2F.