Objecfive To study the effects of some cytokines such as TNF-α,IL-6 and IFN-γ as well as lipopolysaccharide on CD68 expression in HaCaT cells.Methods Human HaCaT keratinocytes were randomly divided into natural proliferation group (without stimulation),IFN-γ-stimulated group,TNF-α-stimulated group,LPS-stimulated group and IL-6 stimulated group.The work concentration of TNF-α,IL-6,IFN-γ and LPS was 50 mg/L.HaCaT cells were collected after 24-hour treatment with the cytokines followed by the examination of CD68 expression with flow cytometry,immunohistochemistry and reverse transcription(RT)-PCR,respectively.Results Compared with untreated HaCaT cells,the count of CD68-positive cells was elevated in cells stimulated by TNF-α(t=3.60,P<0.01),IL-6(t=3.93,P<0.01),IFN-γ(t=2.38,P<0.05)and LPS(t=2.52,P<0.05),and the effect of TNF-α and IL-6 was stronger than that of IFN-γ and LPS.Among the four cytokines,only IL-6 enhanced the mean fluorescence intensity of CD68-positive cells (t=8.34,P<0.01).After 24-hour treatment with TNF-α,IFN-γ and IL-6,CD68 expression was observed in the cytoplasm and on the membrane of HaCaT cells and was stronger in cells treated with TNF-α and IL-6 than in those with the other cytokines.A significant increase was observed in the CD68 mRNA expression after 24-hour treatment with TNF-α (t=4.34,P<0.01),IL-6 (t=7.52,P<0.01)and IFN-γ (t=2.81,P<0.05);TNF-α and IL-6showed a stronger promotive effect than IFN-γ.Conclusion IL-6,TNF-α,IFN-γ and LPS can upregulate the CD68 expression in HaCaT cells.

Objective To establish a reconstructed human epidermis model in vitro, and to study the process of re-epithelialization. Methods A dermal substrate devoid of epidermis was prepared; a 2 mm skin biopsy explant was transplanted onto the dermal substrates. Visualization of epidermal cell migration was carried out by fluorescence imaging. The proliferation and differentiation of the new epithelial cells were observed using histopathological and immunohistochemical staining (Ki67). EGF was added to the culture medium of the experimental samples but not to that of the controls. Results After 3 days of culture, re-epithelialization was observed on the surface of the dermal substrate. A complete structure resembling regular epidermis was noted in 10 days. As compared to control samples, EGF-treated samples had larger area of re-epithelialization (t= 3.02, P

Objective To study the changes in some important surface markers of Langerhans cell (LC) in the skin of SLE patients. Methods The normal-appearing skin and lesional skin of 9 SLE patients were studied. ABC immunohistochemistry and the monoclonal antibodies against HLA-DR, CD54, CD68, CD1a and CD4 were used. Results ①In SLE skin lesions decreased LC density was shown,and there were also changes in the morphology and surface markers of LC. ②HLA-DR expression on keratinocytes was shown in most lesional specimens and a few on non-lesional specimens. ③Neither CD4 nor CD54 expression was shown on both lesional epidermis and non-lesional epidermis, CD4+cells were only observed in the dermal infiltrates. ④Two types of CD68+dendritic cells were seen in lesional and non-lesional epidermis, and more CD68+dendritic cells were seen in the infiltrates of lesional skin. ⑤Fibrillar CD68+materials around the basal KC were observed, and some of such fibrillar materials were connected with epidermal dendritic cells while others were not. Conclusions There are some differences in LC surface marker expression on lesional skin and non-lesional skin of SLE patients, which need to be further studied.

Objective To compare the intensity and isotype of anticardiolipin(ACL)antibodies in the sera from patients with syphilis and SLE.Methods IgG and IgM type of ACL antibodies were examined by ELISA in99syphilis patients and75SLE patients.Results Although similar positive percentage of IgG ACL antibodies was shown in syphilis and SLE patients,stronger absorbance(A value)was seen in syphilis patients(P

Objective To investigate the distribution of antiphospholipid antibodies(APA) in patients with systematic lupus erythematosus (SLE) and normal controls and its significance. Methods Six kinds of APA were detected by enzyme linked immunosorbent assay(ELISA). The examined sera were collected from 82 cases of normal controls and 32 cases of SLE. Results ①The positivity rate of IgG type aCL, aPI, aPA, aPE, aPS and aPC in the serum from normal controls was 6.10%, 6.10%, 7.32%, 7.32%, 3.66% or 3.66%, respectively. ②The positivity rate of IgM type aCL, aPA, aPC, aPS, aPE aPI in serum from normal controls was 2.44%, 3.66%, 3.66%, 6.10%, 6.10% and 4.88%, respectively. ③Compared with normal controls, the A values and positivity rate of IgG type of aCL, aPA, aPS, aPC and aPI in SLE patients were significantly higher. ④Compared with normal controls, the A values and positivity rate of IgM type of aCL, aPA, aPS, aPE and aPC in SLE patients were significantly higher. ⑤The total positivity rate of IgG type and IgM type APA was 68.75% or 71.88%, respectively. Conclusion ①There is lower titer of APA in normal controls, with some relationships between different types of APA and they appear at the same time.②the measurment of APA in sera of SLE might be benefit to the diagnosis of antiphospholipid syndrome. [

Objective To assess the changes in frequency of peripheral T lymphocytes expressing different cytokines in patients with atopic dermatitis (AD) before and after treatment with BCG-PSN and their relationship with disease severity. Methods A randomized, double blinded and placebo cross-over control study was conducted. A total of 8 patients with AD were recruited in this study. Intramuscular BCG-PSN or placebo was given to patients every other day for 36 days. Flow cytometry was performed to measure the frequency of IL4-, IL5-, IFN-γ- and TNFα-expressing peripheral CD4+ T cells and CD8+ T cells before and after the therapy. Disease severity was evaluated by atopic dermatitis area and severity index score (ADASIS). Results The difference value in IFN-γ+CD8+ T cell frequency before and after therapy was significantly higher in patients treated with BCG-PSN than in those with placebo (8.056 ± 13.962 vs -6.549 ± 10.491, U = 2.26, P< 0.05). There was no statistical difference in the frequency of IL4-, IL5-, TNFa-expressing CD8+ T cells between BCG-PSN- and placebo-treated patients (all P > 0.05). The decrease in ADASIS was 1.56 ± 1.49 in patients treated with BCG-PSN, which was statistically higher than that in placebo-treated patients (-0.05 ± 1.54, U = 2.00, P< 0.05). Conclusion As an immunomodulator, BCG-PSN may control AD by restoring the balance of T-cell subsets.

Objective To observe the efficacy of intense pulsed light(IPL)incorporated with optimal pulse technology (OPT)in the treatment of rosacea.Methods Thiris-two patients with erythematotelangiectatic rosacea and 53 patients with papulopustular rosacea were treated with IPL-OPT for 4 sessions with an interval of 3 weeks.Patients were assessed clinically and photographically by physicians before each treatment.Skin melanin index,erythema index,sebum secretion level and water content in stratum corneum were tested at the baseline,3 weeks after each treatment,and 6 months after the last treatment.Results Three weeks after the last treatment,the effective(more than 60%improvement)rate WaS 81.18%in total,75%in erythematotelangiectatic rosacea,and 84.91%in papulopustular rosacea;there WaS no significant difference between erythematotelangiectatic rosacea and papulopustular rosacea (x2=1.28,P＞0.05).Six months after the last treatment,the total effective rate still remained at 78.82%with no significant difierence from that observed at 3 weeks after the treatment(x2=1.62,P＞0.05).The value of melanin index,erythema index and sebum secretion level decreased significantly after the final treatment,however,water content in stratum corneum remained at the salne level as that before treatment.After 6-month follow-up,no significant change was noticed in the above 4 parameters compared with those obtained at 3 weeks after the treatment.Neither hyperpigmentation nor hypopigmentation was observed during the treatment and follow-up.Conclusion This study demonstrates that IPL-OPT is an effective treatment for rosacea with relatively few side effects.

Objective:To observe the distribution,morphology and density of monocytes/macrophages and dendritic cells in the normal human dermis.Methods:Normal skin from 6 locations such as the face,trunk,proximal limbs,distal limbs,and palms and soles of 8 subjects were collected for the study.The horizontal and longitudinal sections of the skin were stained with an ABC immunoperoxidase procedure with anti-CD1a and anti-CD68 monoclonal antibodies.Results:In the superficial dermis CD68 positive monocytes/macrophages form a dense network with a density in a 6-micron section ranging from 361/mm~2 to 562/mm~2.These network of CD68 positive cells continued on to surround the blood vessels and skin appendages.Lower densities of CD68 positive dendritic cells were found in the deep(reticular) dermis,dispersed between collagen bundles.The CD68 positive cells were detected within the superficial dermis with variable densities: distal limbs 562/mm~2,trunk 517/mm~2,face 509/mm~2,palms 507/mm~2,proximal limbs,472/mm~2,and soles 361/mm~2.Conclusion:There exists in the superficial dermis a relatively dense network of CD68 positive monocytes/macrophages.Such a distribution might indicate the clear polarity of the dermal monocytes/macrophages,with their direction of defense towards to the dermal-epidermal junction.

Objective To investigate the role of immunophenotyping in distinguishing mycosis fun- goides (MF) from lichen planus and psoriasis.Methods The expression of CD1a,CD4,CD8,ICAM-1, LFA-1,HLA-DR,CD30 and CD7 was measured by ABC immunohistochemical technique in specimens ob- tained from lesional skin of 15 cases of MF,17 cases of lichen planus and 17 cases of psoriasis,and in the skin of 6 healthy controls.Results In the lesional epidermis of MF,the density of cells positive for CD1a, CD30 or ICAM-1,was significantly higher (mononuclear cells,P＜0.001;dendritic cells,P＜0.01) than that in the lesional epidermis of lichen planus,psoriasis and in the skin of healthy controls.The density of cells positive for CD4 or CD8 and of dendritic cells positive for HLA-DR was higher in lesional epidermis of MF than in that of lichen planus.The linear density of CD1a-positive cells (P＜0.01),the percentages of cells positive for ICAM-1 (P＜0.05) or LFA-1 (P＜0.05) were all higher in the lesional dermis of MF than in that of lichen planus.As far as the CD7-positive cell density was concerned,it was higher in the lesional dermis of lichen planus and psoriasis than in that of MF and skin of healthy controls (P＜0.01), while no difference was found between the epidermis of MF and that of lichen planus or psoriasis.Conclu- sion There are differences in the expression of CD1a,CD4,CD8,ICAM-1,LFA-1,HLA-DR,CD30 and CD7 in the lesional skin of MF,lichen planus and psoriasis,which may provide a clue to the pathogenesis of these diseases.

Objective:To study the effect of astragalus polysacchrides on anticardiolipin, antiphosphatidyl choline, antiphosphatidyl serine, antiphospha-tidyl inositol, antiphosphatidic acid and antiphosphatidyl ethanolamine antibodies (aCL, aPC, aPS, aPI, aPA, aPE) in SLE mice. Methods:19 NZB?NZW F1 female mice were divided randomly into 3 groups: group PG2I (25 mg/kg/d), group PG2II (50 mg/kg/d), group NS (0.2 ml/d). Select BXSB and C57BL/6 female mice as control. The level of antiphospholipid antibodies in these mice were examined by ELISA. Results:Compared with group NS, the A value in group PG2II were decreased significantly, however, A value in group PG2I were elevated slightly. There were no difference among group PG2II, BXSB and C57BL/6. The level of antiphospholipid antibodies in group NS and PG2I were increased significantly compared with that of BXSB and C57BL/6 mice. Conclusion:The generation of antiphospholipid antibodies can be improved by low dose of PG2 and inhibited by high dose of PG2.