Objective To explore the inhibitory effects of He Ne laser repeated irradiation on the collagen synthesis of cultured scar fibroblasts. Method Cultured fibroblasts derived from hypertrophic scars(HS) were irradiated with He Ne laser for 30 minutes at various power densities(10,50,100 and 150 mW/cm2),once a day for 3 consecutive days.In 24 hours after repeated irradiation collagen production and type I procollagen mRNA level of fibroblasts were measured with the incorporation of 3H proline and blot hybridization techniques espectively.Results Collagen synthesis and type I procollagen mRNA level remained unchanged when the laser was irradiated at the power density of 10 mW/cm2 or 50 mW/cm2.Compared with control,collagen synthesis and type I procollagne mRNA level were significantly decreases at the power density of 100 mW/cm2 or 150mW/cm2(P< 0.05).Type I procollagen mRNA level at the power densityof 150 mW/cm2 was lower than that at the 100 mW/cm2 (P< 0.05).Conclusion Repeated He Ne laser irradiation at the power density of 100 mW/cm2 or 150 mW/cm2 can suppress collagen synthesis of cultured fibroblasts in HS.The cause of suppression may be associated with down regulation of type I procollagen mRNA expression.

regulatory T cell(Treg)is a subset of immunosuppressive Th cell lines with ability to sup-press anti-tumor immunity. Tregs are recruited into local tumor microenvironment by tumor-released chemokine CCL22. Tregs induce Th2-type immune responses which contribute the host ignorance and tolerance to tumor cells via releasing of immunosuppressive cytokines and immunosuppressive receptors. Accumulating evidences indicate that inhibiting Treg can break immune tolerance,and subsequently inhibit the growth and progress of cancer. These discoveries and exploration of Treg provide us a new therapeutic target and strategy with promising clinic application.

Cytotoxic T lymphocytes (CTLs) are critical effectors which play important roles both in anti-tumor and anti-virus immune responses.Through T cell receptors(TCRs),CTLs can specifically recognize MHC- Ⅰ -peptides complexes presented on the surface of target cells,and then release biological substances such as perforin and granzymes into the target cells and dissolve them.Since the significant potential value of CTLs,more and more people are focusing on its basic and appliedstudies.Most researches are about the transformation of the TCR gene.With the development of molecular biology,cloning and transduction of TCR gene have been more mature.Researchers are finding ways to ensure that TCR gene expresses efficiently and assemble correctly.

Objective:To express recombination human interleukin-13(rhIL-13)in methylotropic yeast Kchia pastoris and study its effect on the induction of DC from PBMC in vitro. Methods: An artificial gene with the optional condon usage of Kchia pastoris for IL-13 was designed and synthesized by T4 DNA ligase and PCR.The expression vector pPICZoA-IL-13 was constracted and introduced in Kchia pastoris GS115 by electroporation after linearized with Sacl.Recombinants were selected by plating cells on YPD/Zeocine plates.The recombinant protein secreted by the yeast was identified by 15%SDS-PAGE,ELISA and Western blot.The abilities to induce DC differentiation of IL-13 and IL-4 were compared. Results: Based on the result of the 15% SDS-PAGE and Western blot assay, an about 13 kD recommbinant protein expressed in the yeast supernatant was identified to be recombinant human IL-13. Adherent PBMC cultured in GM-CSF,TNFa and IL-13 displayed morphological characteristic of DC generated in media containing IL-4. They formed cellular clumps and had extensive dendrites sharp. Fluorescence-activated cell sorter analysis showed that they expressed a high level of MHC class II ,CDla,CD80,CD83,CD86. The purity and yield of both DC populations are not significantly different with IL-13 or IL-4. The phagocytosis of immature DC induced by IL-13 is potent. They were also equally potent stimulators of allogeneic lymphocytes in the mixed lymphocyte reaction. Conclusion: rhIL-13 can substitute for IL-4 in the proliferation and development of dendritic cells.

Objective To study the immuno-regulatory roles of antibiotics and their implication for pulmonary fibrosis.Methods The regulatory roles of antibiotics on the phenotypes of dendrite cells(DCs) and maturation of B lymphocytes in vitro were detected with flow cytometry.The roles of nystatin on pulmonary fibrosis in vivo were determined with bleomycin-induced acute injury model of mouse.Results Multiple antibiotics including nystatin apparently promoted the maturation of DCs and B cells,as well as subsequently promoted the polarization of Th2 cells through DCs.In addition,nystatin intensified bleomycin-induced pulmonary fibrosis.Conclusion Multiple antibiotics including nystatin induce suppressive immune responses and might aggravate pulmonary fibrosis like nystatin.

[ABSTRACT]OBJECTIVETo investigate the effect of accurate airway humidification on hemorrhage, pharyngalgia, mucosal edema and sputum viscosity in patients with low-temperature plasma coblation-assisted tonsillectomy.METHODS58 cases were divided into three groups by using random numbers.In accurate airway humidification group, atomizing inhalation was carried out by AIRVOTM series apparatus; in oxygen atomizing group, budesonide suspension was used; in control group, saline was used. We evaluated the hemorrhage, pharyngalgia, mucosal edema and sputum viscosity in 3 consecutive postoperative days.RESULTSPharyngalgia in accurate airway humidification group and in oxygen atomizing group were both significantly reduced than that of the control group (P<0.001). Besides, in accurate airway humidification group, mucosal edema and sputum viscosity were significantly improved than that of the oxygen atomizing group (P<0.05) and control group (P<0.05).CONCLUSIONAccurate airway humidification could reduce the complications such as pharyngalgia, mucosal edema and purulent sputum after low-temperature plasma coblation-assisted tonsillectomy, and could accelerate recovery from surgery.

Objective To investigate the safety and efficacy of CO2 fractional laser combined with 5-aminolevulinic acid (ALA)-mediated photodynamic therapy (ALA-PDT) for the treatment of refractory flat wart.Methods The 80 patients with facial flat wart were randomly divided into observation group (40 cases) and control group (40 cases).The observation group was treated with CO2 fractional laser combined with 5-ALA-PDT,and the control group was treated with 5-ALA-PDT only.Those with residual lesions after the first treatment were treated with the second or third times.The interval duration between treatments was 2 weeks.The efficacy and the adverse reaction were evaluated after the 2-week,4-week and 6-month after the last treatment.Results Compared with prior treatment,the facial lesions achieved complete response and improvement.The total effective rate of the observation group were 90.0％ (36/40) and 72.5％ (29/40) for the control group,the difference was statistically significant (P＜0.05).There were 2 cases of pigmentation occurred in observation group and 3 cases in control group.The complications including infection and hypertrophic scar were seldom occurred in the two groups.Conclusions The safety and efficacy of CO2 fractional laser combined with ALA-PDT are better than that with ALA-PDT only in the treatment of refractory flat wart.

Objective To evaluate the role of interleukin-4 receptor (IL-4R) in renal fibrosis following renal ischemia-reperfusion (I/R) injury in mice.Methods Twelve male wild type BALB/C mice and 12 IL-4Rα gene-knockout mice,aged 8-10 weeks,weighing 20-30 g,were used in the study.The mice of either type were divided into 2 groups (n =6 each) using a random number table:sham operation group (group S) and group I/R.In group I/R,renal I/R was induced by occlusion of the right renal artery for 1 h with atraumatic microclips followed by 2 weeks of reperfusion.The right renal artery was only isolated in group S.At 2 weeks of reperfusion,blood samples were taken from the orbital vein for determination of the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr).The renal tissues were obtained,and the renal fibrosis area was measured by Sirius Red staining.The expression of fibronectin (FN),collagen Ⅰ (COL-Ⅰ) and α-smooth muscle actin (α-SMA) in renal tissues was detected by immunofluorescence.The expression of signal transducer and activator of transcription 6 (STAT6) and phospho-STAT6 in renal tissues was determined by Western blot.The ratio of phoshop-STAT6 to STAT6 was calculated to reflect the phosphorylation of STAT6.Results Compared with group S of wild type mice,the serum BUN and Cr concentrations and renal fibrosis area were significantly increased,the expression of FN,COL-Ⅰ and α-SMA in renal tissues was significantly up-regulated,and the phosphorylation of STAT6 in renal tissues was significantly increased in group I/R of wild type and IL-4Rα KO mice (P＜0.05).Compared with group I/R of wild type mice,the serum BUN and Cr concentrations and renal fibrosis area were significantly decreased,the expression of FN,COL-Ⅰ and α-SMA in renal tissues was significantly down-regulated,and the phosphorylation of STAT6 in renal tissues was significantly decreased in group I/R of IL-4RαKO mice (P＜0.05).Conclusion The mechanism of renal fibrosis following renal I/R injury is partially related to IL-4R,and IL-4R results in renal fibrosis through promoting activation of STAT6 signaling pathway in mice.

Objective To evaluate the effect of propofol on invasiveness of human gastric cancer MKN-45 cells.Methods Human gastric cancer cell line MKN-45 were seeded in culture plates.After being cultured for 24 h,the cells were randomly divided into 5 groups(n =12 each):control group (group C),intralipid group (group Ⅰ),4 μg/ml propofol group (group P1),8 μg/ml propofol group (group P2) and 16μg/ml propofol group (group P3).The cells were treated with 10％ intralipid and 4,8 and 16 μg/ml propofol for 24 h in I and P1-3 groups,respectively.The cells were then cultured for another 24 h.The migration of cells was determined by cell scratch test.The invasion of cells was determined by Transwell invasion assay.The expression of RhoA and ROCK1 was detected by Western blot.Results Compared with group C,the cell migration and invasion were significantly decreased,and the expression of RhoA and ROCK1 was down-regulated in P1-3 groups,and no significant changes were found in the parameters mentioned above in group Ⅰ.With the increasing concentrations of propofol,the cell migration and invasion were gradually decreased,and the expression of RhoA and ROCK1 was gradually down-regulated in P1-3 groups.Conclusion Propofol can inhibit the invasiveness of human gastric cancer MKN-45 cells cultured in vitro dose-dependently and inhibition of RhoA/ROCK1 signaling pathway may be involved in the mechanism.

Objective To preliminarily evaluate the role of Ras homolog gene (Rho)/Rho-associated coiled coil-forming protein kinase (ROCK) signaling pathway in propofol-induced inhibition of metastasis of human gastric cancer cells.Methods Human gastric cancer MKN-45 cells cultured in vitro,with the concentration of 1.0× 106 cells/ml,were seeded in culture plates,and incubated for 24 h.The plates were then randomly divided into 4 groups using a random number table:control group (group C),propofol group (group P),lysophosphatidic acid group (group L) and propofol + lysophosphatidic acid group (group PL).Group C received no administration.In group P,propofol at the final concentration of 16 μg/ml was given.In group L,lysophosphatidic acid at the final concentration of 1 μmol/L was administered.In group PL,propofol and lysophosphatidic acid were given with the final concentration of 16 μg/ml and 1 μmol/L,respectively.All the cells were then incubated for another 24 h.The migration of cells was determined by wound healing assay,and cell migration rates were calculated.The invasion of cells was determined by Transwell assay,and the invaded cells were counted.The expression of matrix metalloproteinases-2 (MMP-2),MMP-9,RhoA,and ROCK1 in cells was detected by Western blot.Results Compared with group C,cell migration rates and the number of invaded cells were significantly increased,and the expression of MMP-2,MMP-9,RhoA and ROCK1 was up-regulated in group L,and cell migration rates and the number of invaded cells were decreased,and the expression of MMP-2,MMP-9,RhoA and ROCK1 was down-regulated in group P.Compared with group L,cell migration rates and the number of invaded cells were significantly decreased,and the expression of MMP-2,MMP-9,RhoA and ROCK1 was down-regulated in group PL.Conclusion Inhibition of RhoA/ROCK1 signaling pathway is involved in the mechanism by which propofol decreases metastasis of human gastric cancer cells.