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Objective To investigate the suppressive effect of prostaglandin E2 (PGE2),hepatocyte growth factor (HGF) and indoleamine 2,3-dioxygenase (IDO) whose secretion was promoted by human umbilical cord mesenchymal stem cells(MSCs) on the activated peripheral blood CD4+ T cells in primary Sj(o)gren syndrome (pSS) in vitro.Methods Primary cultured umbilical cord MSCs were identified by flow cytometry,and peripheral blood CD4+ T cells were sorted in pSS patients.CD4+ T cells were cultured with CD3,CD28 antibody for 72 h to be the activated(control group);the activated CD4+ T cells were co-cultured with MSCs for 72 h(MSCs group) or MSCs were pre-stimulated with interferon-γ (IFN-γ),then the activated CD4+ T cells were co-cultured with pre-stimulated MSCs for 72 h (pre-stimulated group).The suspension of CD4+ T cells were collected and counted.PGE2,HGF and IDO in the supernatants were detected by ELISA.Mean in groups were compared using ANOVA,and multiple comparisons were used with LSD method.Results The concentrations of PGE2 in the supernataut of the control group,MSCs group and pre-stimulated group were (111 ±4) pg/ml,(2 814±6) pg/ml and (2 716±8) pg/ml (F=167 292.12,P＜0.01) respectively.The concentrations of HGF in the above groups were (597±9) pg/ml,(383±9) pg/ml and (727±12) pg/ml(F=878.61,P＜0.01) respectively.The concentrations of IDO in the above groups were (143±4) pg/ml,(835±5) pg/ml and (588±3) pg/ml (F=21 104.41,P＜0.01) respectively.Compared with the control group,levels of PGE2 significantly increased in the MSCs group and the pre-stimulated group that CD4+ T cells were co-cultured with MSCs (t=509.88,P＜0.01 and t=491.48,P＜0.01),and levels of IDO also significantly increased (t=202.69,P＜0.01 and t=130.39,P＜0.01),while the activation and proliferation of CD4+T cells were inhibited (t=-16.20,P＜0.01 and t=-31.48,P＜0.01).Compared with MSCs group,levels of PGE2 and IDO significantly decreased in pre-stimulated group (t=-18.40,P＜0.01 and t=-72.30,P＜0.01),and levels of HGF significantly increased in pre-stimulated group(t=41.51,P＜0.01),while the activation and proliferation of CD4+ T cells were further inhibited (t=-15.28,P＜0.01).Conclusion MSCs can inhibit the activation and proliferation of CD4+ T cells in pSS in vitro.The suppressive effect of MSCs may be achieved by promoting secretion of cytokines such as PGE2,HGF and IDO.HGF plays a more important role in the suppressive effect of MSCs pre-stimulated with IFN-γ.Too much PGE2 or IDO propably results in negative feedback regulation of MSCs.
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Objective To explore the relationship between the clinical features,serological markers and European League Against Rheumatism SS Disease Activity Index (ESSDAI) scores of primary Sj(o)gren's syndrome (SS).Methods We enrolled 106 patients,who fulfilled the 2002 classification criteria for primary SS from December 2008 to January 2015,to evaluate the relationship among the clinical characteristics,laboratory features,serological variables and ESSDAI scores.According to serological variables,the prognosis was subdivided into three distinct groups:favourable (no serological markers),intermediate (one serological marker) and poor (two or more serological markers).These data were analyzed by Chi-square test and variance analysis.Results The mean ESSDAI score of 106 pSS patients was (11±7).ESSDAI score was categorized according to the EULAR-SS recommendations as low activity,moderate activity and high activity (scores of 0-4,5-13 and ≥14,respectively),and the positive rate of antinuclear antibody (ANA) 1:100 (6 cases,37.5％;37 cases,66.1％;32 cases,94.1％) in three different ESSDAI levels was statistically different (x2＝18.110,P＜0.01).Those with positive ANA 1:100[positive (13±7) and negative (7±4)],anti-SSA antibody postive (12±7) and negative (9±7),anti-RNP antibody (positive 16±9 and negative 10±6) had higher ESSDAI scores than those with negative ones (F=8.812,P=0.0001;F=3.862,P=0.024;F=5.786,P=0.004).No statistical difference in ESSDAI means were found between patients with positive anti-SSB antibody,rheumatoid factor (RF),FS level,dry mouth,Raynoud's phenomenon and psychosomatic diseases.The ESSDAI scores of favourable group,intermediate group and poor group were significantly different (8±5,10±7,14±7,F=8.715,P=0.000 1).In comparison with the other two groups,the poor pSS patients had a higher frequency of positive ANA 1:100 (15 cases,55.6％;20 cases,57.1％;40 cases,90.9％),anti-SSA antibody(11 cases,0.7％;23 cases,41.1％;36 cases,81.8％),anti-SSB antibody (6 cases,2 2.2％;13 cases,37.1％;23 cases,52.3％),anti-RNP antibody (0 case,0;2 cases,5.7％;9 cases,20.5％) (x2=17.408,P=0.002;x2=14.306,P=0.006;x2=12.330,P=0.015;x2=1 1.482,P=0.022).Conclusion Patients with two or more serological markers may have higher ESSDAI score,and which in turn may associate with poor prognosis.
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<p><b>OBJECTIVE</b>To explore the role of ompT gene in uropathogenic E. coli (UPEC) CFT073 strain in urinary tract infection (UTI).</p><p><b>METHODS</b>An ompT deletion mutant (COTD) was generated by λ Red recombineering in the UPEC CFT073 strain, which was characterized by PCR and sequencing. C57B/L6 mouse models of acute UTI with the mutant and wild-type strains were established to compare the colonization abilities of the two strains in the bladder. The adhesion of CFT073 mutant to human unthelial 5637 cells was also investigated in vitro.</p><p><b>RESULTS</b>PCR and DNA sequencing confirmed the loss of ompT gene in the mutant COTD. The in vitro adhesion rate of the mutant strain COTD to 5637 cells was (6.7±2.2)%, significantly lower than that of (8.3±1.9)% of the wild-type strain (P<0.05). In the murine models of acute UTI, the mutant strain showed a mean colonization number of about (17±8)×10⁴ cfu, which was significantly lower than that of (7∓2)×10⁵ cfu of the wide-type CFT073 strain (P<0.05).</p><p><b>CONCLUSION</b>OmpT gene can be involved in the colonization of UPEC in the bladder tissue and plays an important role in the pathogenesis of UPEC-induced UTI.</p>