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Objective To investigate the influencing conditions of human epidermal stem cells(ESCs) differentiating into gland-like cells and to identify the induced tubular structure.Methods The ESCs were seeded onto compound polysaccharide shell dermal matrix and collagen gel,adding different concentrations of epidermal growth factor(EGF) and culturing by vertical shaking in vitro,the three dimensional culture and induced directional differentiation were performed.The means of HE staining,immunofluorescence and confocal laser scanning microscope were adopted to observe the conditions,morphology and phenotype change of ESCs directionally differentiating into glandular epithelium-like cells.Results 15-20 ng/mL EGF could induce ESCs in tissue-engineering dermis to grow into dermis and appear the gland-like structure.The HE staining in this structure showed its profile as a single layer with lacune in the middle compartment,eosinophilic cytoplasm,lightly stained.Under CLSM,by CK19 staining,the luminal structure in the middle of the cell mass was observed,while CK18 and CEA were expressed in this structure.Conclusion Under the induction of particular concentration of EGF,the in vitro cultured human ESCs seeded on the tissues engineering dermis could form into the tubular structure,which is similar to the in vivo sweat gland secretory cells in morphology and histology.
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Objective To establish the three-dimensional culture model of the human skin squamous-celled carcinoma ,and isolate the highly invasive cell subsets ,and compare their biological characteristics .Methods By using the composite chitosan tissue engi-neering skin ,the three-dimensional culture model of the human skin squamous-celled carcinoma was successfully established , through digestion ,we divided and got the high-invasive squamous carcinoma cells subsets .After that ,the different invasive proper-ties of subpopulations of cells in nude mice tumor effects were compared .Results Different concentrations of cinnamaldehyde and interferon on skin squamous-celled carcinoma cell proliferation inhibition had statistical significance (P<0 .01) ,the different invasive characteristics of skin squamous cells carcinoma between subpopulations of cells proliferation inhibition also was statistically signifi-cant(P<0 .01) ,and the two different invasive properties of subpopulations of cells in the tumor characteristics also had statistical significance(P<0 .01) .Conclusion We established the three-dimensional culture model of the human skin squamous-celled carcino-ma successfully ,by using this model ,we could screen the higher invasive cells subsets .
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Objective To detect the expression and function of cysteinyl leukotriene receptors (CysLTRs)in keratinocytes.Methods Human keratinocytes were isolated from the tissue of foreskin by digestion with dispase Ⅱ and trypsin,and subjected to primary culture.By using confocal laser scanning microscopy and reverse transcriptase PCR,the localization and expression of CysLTRs were studied in kemtinocytes.respectively.Some primarily cultured keratinocytes were pretreated with leukotriene D4 (30 nmoi/L),MK571(300 nmol/L),and BAYu9773 for 5 minutes followed by the detection of intmcellular calcium level using the Ca2+ indicator dye Fura-2/AM as well as cell proliferation bv MTT assay.Results The expressions of CysLTR1 and CysLTR2 were observed in cultured keratinocytes,and they were mainly located on cell membrane,partly in cytoplasm and nuclei.Compared with non.stimulated cells,a significant increase Was noted in the expression of CysLTRs,especially in the nuclei of keratinocytes stimulated by LTD4(P＜0.05),together with an elevation in intracellular calcium level(42.27±3.00 mmol/L,P＜0.01)and acceleration in cell proliferation (P＜0.01).However,both MK571 and BAYu9773 could completely block the effect of LTD4 on intmcellular calcium level and cell proliferation.and there was no significant difference in the blocking effect between MK571 and BAYu9773.Conclusions Functional CysLTRs are expressed in human keratinocytes.and they carl increase the intracellular calcium level in,and cell proliferation of,keratinocytes.
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0.05).Conclusion The improved device is suitable for detecting the tension strength of small tension membranaceous biomaterials such as tissue engineered skin.
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Objective To report the diagnosis and treatment of the pregnant women, who were positive for syphilis in serological test, as well as their final pregnant results. Methods we conducted a retrospective analysis of the serological test results of 3 105 pregnant women subjected to childbirth or induced labor in the department of gynaecology and obstetrics at our hospital since January of 2004. Results Of the 3105 pregnant women, 21 (6.76 ) were positive for treponema pallidum (TP). Additionally, 7 (33.33%) cases were positive for TRUST, and 6 (28.57%) negative. Only 2 cases showed skin lesion, canker and soft chancre. Of the 18 cases, who were found to get infection during pregnancy, 11 (61.11%) had taken the treatment against syphilis. Of the 21 women positive for TP, 5 were subjected to induced labor, 13 to cesarean section, and 3 to spontaneous parturition. The average time of pregnancy was (267.44?11.00) d. The Apgar score of all 16 newborns was 10 at birth. The umbilical blood of 5 infants was tested by TP and TRUST, among which 3 were positive by TP, 2 were positive for TRUST. All infants did not have any clinical manifestation of congenital syphilis after birth. There were 5 cases transferred to the ward of pediatrics, among which 2 undertook the treatment against syphilis, 2 against premature delivery, and 1 against asphyxia, and finally all had been cured. Conclusion The patients, who were found positive for TP during the early and middle period of pregnancy, can obtain a fine pregnant result, if undertaking regular examination and treatment against syphilis.
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Objective To study the effects of recombinant chimeric toxin Dsg3EC_(1-2)PE40 on T and B cells from PV patients. Methods The recombinant protein Dsg3EC_(1-2)PE40 was expressed on BL21TrxB (DE3) cells, then identified and purified. ELISPOT assay was used to detect and quantitate autoantibody-producing B cells in different concentrations of recombinant chimeric toxin, and MTT assay and ~3H-TdR assay to observe the metabolism and proliferation of T cells from PV patients in vitro. Results The purity of expressed protein Dsg3EC_(1-2)PE40 was up to 80%. The number of anti-Dsg3 antibody-producing B cells in PBMC from PV patients decreased by 40% with treatment of Dsg3EC_(1-2)PE40, which was significantly lower (P
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Objective To assess the levels of nucleosomes released from peripheral blood mononu-clear cells(PBMC)of patients with systemic lupus erythematosus(SLE),and their relationship with auto-antibodies as well as disease activity.Methods Levels of both nucleosomes released from PBMC and vari-ous auto-antibodies were detected by ELISA in sera from SLE patients.The disease severity was evaluated using SLEDAI(systemic lupus erythematosus disease activity index)system.Results Levels of nucleosomes released from PBMC were significantly higher in patients with active SLE than those of patients with inactive disease and normal controls(39.39?25.70,13.44?8.82,and11.73?7.87IU/mL,respectively).There was a significant positive correlation between nucleosome levels and SLEDAI scores,serum ds-DNA auto-an-tibody levels,and low C3levels.Conclusion Nucleosomes released from apoptotic PBMC of patients with SLE is closely correlated to disease activity,which implies that nucleosomes may play an important role in the pathogenesis of SLE.
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Objective To investigate the role of cytotoxic T lymphocyte associated molecule-4Ig(CTLA-4Ig) in the prevention of C57BL/6 mice autoimmune hepatitis. Methods The C57BL/6 mice were intraperitoneally immunized with C57BL/6 mice liver-specific protein in complete Freund's adjuvant. At the same time CTLA-4Ig were given to observe the pathologic alteration of C57BL/6 mice liver. Results With the increase of time of immunization, the results in the treatment group were similar to those of the control group; but inflammatory cell infiltration, hepatic cell swelling, focal necrosis and severe hepatocyte damage were found in the pathologic model group. There was a significant difference between the pathologic model group and control one. Conclusion Autoimmune hepatitis of C57BL/6 mice can be effectively prevented by CTLA-4Ig.
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Objective To investigate the actions of extra cellular medium in growth and differentiation of hair follicle and to look for growth adjusting factors for dermal papilla cells (DPC). Methods Dermal papilla cells were isolated and cultivated with two steps method and the cells were identified by immunohistochemical staining for actin. Influence was examined on the adhesion and growth of dermal papilla cells by chondroitin sulfate A, chondroitin sulfate C and heparin sulfate. Results Two steps method of enzyme digestion for isolating and cultivating dermal papilla cells was an efficient method and large amount of dermal papilla of high purity were harvested with this method. The method is very simple and easy to manege with. Increased adhesion and growth of dermal papilla cells were observed in specimen treated with chondroitin A and heparin sulfate. No significant effects was observed in the cells treated with chondroit in sulfate C. Conclusion Some extra cellular medium can regulate the adhesion and growth of dermal papilla cells and therefore influence the growth and development of hair follicle.
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Objective To investigate the role of nucleosome in the pathogenesis of systemic lupus erythematosus (SLE). Methods BALB/c mice were immunized with nucleosome, and then serum dsDNA and ANA autoantibodies were detected by ELISA. Kidney specimens were observed by immunofluorescence and histological examination. Results High titers of IgG dsDNA and ANA autoantibodies in sera of BALB/c mice were observed at the 14th day after immunization with nucleosome. Nephritis and immune complex deposition in renal glomeruli were observed at the 35th day. Conclusion Nucleosome could induce SLE-like disease in BALB/c mice, and may play a critical role in the pathogenesis of SLE.