Objective To detect the expression level of CXCL9 and IL‐22 in peripheral blood of patients with vitiligo vulgaris and explore its role in the pathogenesis of vitiligo vulgaris .Methods The level of CXCL9 and IL‐22 in peripheral blood from 35 vit‐iligo patients (20 active cases and 15 stable cases) and 20 normal controls were measured by enzyme linked immunosorbent assay . Then ,we compared the differences in different groups and analyzed their correlation with ages ,duration ,psoriasis area and severity index (PASI) .Results The contents of CXCL9 and IL‐22 in active cases and stable cases were obviously higher than those in con‐trols respectively by statistical analysis ,there was significant difference (P< 0 .05) .The contents of CXCL9 and IL‐22 in active ca‐ses were obvious higher than those in stable cases ,there was significant difference(P< 0 .05) .Conclusion CXCL9 and IL‐22 may be involved in the pathogenesis of vitiligo vulgaris .

Objective To investigate the expression and significance of CXCL 9 in systemic lupus erythematosus (SLE ) . Methods The level of CXCL9 in peripheral blood from 20 patients with SLE and 20 normal controls were measured by enzyme linked immunosorbent assay .Compare the difference of the expression level of CXCL 9 in peripheral blood between two groups and analyzed their correlation with ages ,duration ,SLE diseases activity index (SLEDAI) .Results The level of CXCL9 in peripheral blood in patients with systemic lupus erythematosus was (1 549 .14 ± 362 .74)pg/L ,but in normal controls was(602 .54 ± 83 .70)pg/L .The level of CXCL9 in peripheral blood in patients with systemic lupus erythematosus was obviously higher than that in controls by statistical analysis ,there was significant difference between the two groups (P<0 .01) .The expression level of CXCL9 in pe‐ripheral blood of patients with systemic lupus erythematosus was positively correlated with SLEDAI (r=0 .892 ,P<0 .01) .Conclu‐sion CXCL9 may be involved in the pathogenesis of systemic lupus erythematosus .

Objective To detect the expressions of CD70 mRNA and protein and to determine the methylation status of CD70 gene promoter in T cells from patients with systemic lupus erythematosus (SLE).Methods Peripheral blood CD4+ and CD8+ T cells were isolated from 15 patients with active SLE,15 patients with inactive SLE and 15 healthy control subjects.Real-time quantitative reverse transcription-PCR was carried out to quantify the mRNA expression of CD70,flow cytometry to determine the frequency of CD4+CD70+ and CD8+ CD70+ T cells,and bisulfite sequencing to evaluate the methylation status of CD70 gene promoter in CD4+ and CD8+ T cells.Differences in these parameters among these groups were analyzed by one-factor analysis of variance and SNK-q test.Results Compared with the healthy controls,the patients with active SLE and inactive SLE showed a significant increase in CD70 mRNA expression in CD4+ T cells (0.82 ± 0.12 and 0.73 ± 0.11 vs.0.45 ±0.09,F =53.017,P ＜ 0.01) and in the frequency of CD70+CD4+ T cells (80.30％ ± 11.04％ and 66.80％ ± 3.98％ vs.12.48％ ± 3.45％,F =311.517,P ＜ 0.01).Also,the expression of CD70 mRNA in CD4+ T cells and the frequency of CD70+CD4+ T cells were significantly higher in patients with active SLE than in patients with inactive SLE (both P ＜ 0.05).There was a positive correlation between the frequency of peripheral CD70+CD4+ T cells and disease activity in SLE in these patients (r =0.792,P ＜ 0.01).The average methylation index of the region between-600 bp and-300 bp of CD70 gene promoter in CD4+ T cells was 0.32 ± 0.05 and 0.36 ± 0.05 respectively in the patients with active and inactive SLE,significantly lower than that in the healthy controls (0.62 ± 0.05,F =152.64,P ＜ 0.01),and the patients with active SLE showed a significantly lower methylation index than those with inactive SLE (P ＜ 0.05).Conclusions The CD70 gene promoter in CD4+ T cells is significantly hypomethylated in patients with SLE,which may directly lead to the overexpression of CD70.

BACKGROUND:Research on ethyl pyruvate detection methods is reported rarely, and moreover, literature about reversed-phase high-performance liquid chromatography (RP-HPLC) for detection of ethyl pyruvate is less.
OBJECTIVE:To establish an RP-HPLC method for determination of ethyl pyruvate in ethyl pyruvate-chitosan nanoparticles.
METHODS: The chromatographic analysis was performed on a ZORBAX Eclipse XDB-C18 column (4.6 mm× 150 mm, 5μm) at 25℃, with the mixture of acetonitrile and water (40:60, V/V) as the mobile phase at the flow rate of 1 mL/min. The determination wavelength wasset at 210 nm and the injection volume was 20 μL.
RESULTS AND CONCLUSION: The peak of ethyl pyruvate and the peaks of auxiliary materials and solvent were separated wel. The linear rang of ethyl pyruvate was 1-100 mg/L (r=0.999 6). The relative standard deviation of both the intra-and inter-day precision was less than 3% for low-, moderate-, and high-concentration ethyl pyruvate. The relative standard deviation of reproducibility test and stability test was 1.25% and 1.3%, respectively. Sample average recovery rates were (91.5±1.0)%, (3.5±0.2)%, (94.4±0.4)%, respectively. Encapsulation efficiency of samples were (87.2±0.22)%, (90.5±0.15)%, (91.1±0.17)%, respectively. The relative standard deviation of different sample content were 0.9%, 0.5%, 0.3%, respectively. The RP-HPLC method for determination of ethyl pyruvate is sensitive, accurate and highly specific with wide linear range and high sample average recovery.

Objective To investigate the association of tumor necrosis factor-ot (TNF-α) gene promoter polymorphisms with generalized pustular psoriasis.Methods Totally,91 patients of Han nationality with generalized pustular psoriasis (generalized pustular psoriasis group) and 102 health checkup examinees (healthy control group) were enrolled into this study.PCR and direct sequencing were performed to analyze the-238,-308 and-857 polymorphic sites of the TNF-α promoter.Results The frequency of the A allele at TNF-α-238 site was significantly higher in the generalized pustular psoriasis group than in the healthy control group (P =0.003,OR =4.819,95％ CI:1.581-14.694),so was the frequency of GA/AA genotype (P =0.006,OR =4.455,95％ CI:1.410-14.077).However,no significant differences were observed in the frequencies of G/A alleles (P =0.794) and GG/GA/AA genotypes (P =0.786) at TNF-o-308 site,or in the frequencies of C/T alleles (P =0.474) and CC/CT/TT genotypes (P =0.453) at TNF-α-857 site,between the generalized pustular psoriasis group and healthy control group.Conclusion TNF-α-238G ＞ A polymorphisms may be associated with the occur-rence of generalized pustular psoriasis.

BACKGROUND:Astrocytes play an important role in the pathology of central nervous system diseases.The phenotypic and functional changes in astrocytes suggest that it may be an effective therapeutic target for central nervous system diseases.Our previous studies have confirmed that astragaloside can inhibit the lipopolysaccharide-induced astrocyte inflammatory response.Whether astragaloside can regulate the phenotype and function of astrocytes through Notch-1 and its downstream signaling pathway remains unclear. OBJECTIVE:To explore the effect of astragaloside on astrocyte activation and inflammatory response induced by inflammation and its possible mechanism. METHODS:Cerebral cortex astrocytes derived from neonatal C57BL/6 mouse were cultured in vitro.CCK-8 assay was used to determine the optimum concentration of astragaloside and Notch active inhibitor DAPT.The astrocytes were divided into five groups:PBS group,lipopolysaccharide group,lipopolysaccharide + astragaloside group,lipopolysaccharide + DAPT group and lipopolysaccharide + DAPT + astragaloside group.The secretion level of inflammatory factors was detected by ELISA,and the level of nitric oxide was detected by Griess method.The astrocytes and splenic mononuclear cells were co-cultured in Transwell chamber to observe the migration of CD4T cells.The expression of astrocyte activation marker GFAP,A1 marker C3 and A2 marker S100A10 as well as Notch 1 and Jag-1 was detected by immunofluorescence staining.The expressions of CFB,C3,S100A10,PTX3,Notch-1,Jag-1,and Hes were detected by western blot assay. RESULTS AND CONCLUSION:(1)According to the results of CCK8 assay,the final concentration of astragaloside was selected as 25 μmol/L and the final concentration of DAPT was 50 μmol/L for follow-up experiments.(2)Compared with PBS group,interleukin-6,interleukin-12 and nitric oxide secretion levels in the lipopolysaccharide group were significantly increased(P<0.05,P<0.05,P<0.01).Compared with the lipopolysaccharide group,interleukin-6(all P<0.05),interleukin-12(P>0.05,P<0.05,P<0.05)and nitric oxide(P<0.05,P<0.01,P<0.01)secretion significantly reduced in the lipopolysaccharide + astragaloside group,lipopolysaccharide +DAPT group,lipopolysaccharide + DAPT + astragaloside group.(3)Compared with the PBS group,the expression of GFAP that is the marker of activated astrocytes and the migration of CD4 T cells were significantly increased in the lipopolysaccharide group(P<0.01).Compared with the lipopolysaccharide group,astrocyte activation was significantly inhibited and CD4 T cell migration was significantly reduced in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group(P<0.05,P<0.05,P<0.01).(4)Compared with the PBS group,the expressions of A1 markers C3 and CFB in the lipopolysaccharide group were increased(P<0.01,P<0.05),and the expressions of A2 markers S100A10 and PTX3 were decreased(P<0.01,P<0.05).Compared with the lipopolysaccharide group,C3(all P<0.01)and CFB(both P<0.05)were significantly reduced and S100A10(all P<0.01)and PTX3(P<0.05,P<0.05 and P>0.05)were increased in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group.(5)Compared with the PBS group,the expressions of Jag-1,Notch-1 and Hes in the lipopolysaccharide group were significantly increased(all P<0.01).Compared with the lipopolysaccharide group,the expressions of Jag-1(all P<0.01),Notch-1(all P<0.01)and Hes(P<0.05,P<0.01 and P<0.01)were significantly reduced in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group.(6)The results indicate that astragaloside can promote the transformation of astrocytes from A1 to A2 by regulating Notch-1 signaling pathway,reduce the secretion of inflammatory factors and the migration of CD4 T cells,and thus inhibit astrocyte activation and inflammatory response.