[Terminal-restriction fragment length polymorphism analysis of diversity of mucosa-associated microbiota in patients with Crohn's disease].

OBJECTIVE: To explore the differences of colonic mucosal-associate bacterial diversity between the patients with Crohn's disease (CD) and healthy controls. METHODS: Eight CD patients and 23 healthy controls were recruited from September 2010 to December 2011 at West China Hospital. One biopsy were taken from cecum of every patient with CD and healthy control by endoscopic examination. The diversity of colonic mucosa-associated microbiota was detected by terminal-restriction fragment length polymorphism (T-RFLP) . Hierarchical cluster analysis were performed to compare the similarity of microbial communities between CD patients and healthy controls. Differences of bacterial diversity between two groups were also evaluated. The difference of predominant terminal-restrict fragments (T-RF) were analyzed and the bacterium predicted by predominant T-RFs were identified according to MiCA database. RESULTS: Hierarchical cluster analysis showed that the mucosal microbial community of CD patients differed from healthy controls. And there were more similarities in the samples of same group than that of different groups. Compared with healthy control group, the richness of mucosal microbiota in CD patients was lower (HaeIII:7 ± 4 vs 10 ± 8, P = 0.048; MspI+HaeIII:20 ± 10 vs 24 ± 12, P = 0.036). Shannon-Wiener index of CD patients was lower than healthy control (1.7 ± 0.7 vs 2.0 ± 0.5, P = 0.220) with no significant difference. Species evenness and Simpson index of CD patients were significantly greater than healthy controls (0.84 ± 0.14 vs 0.77 ± 0.13, P = 0.045; 0.25 ± 0.16 vs 0.22 ± 0.15, P = 0.038) . The T-RF of 37, 40 and 66 bp digested with MspI enzyme predominated in CD patients. Relative quantitative analysis showed 35 bp T-RF digested with MspI was significantly higher in CD patients than that in healthy controls (36.8% (23.0%, 55.4%) vs 14.3% (9.5%, 19.5%), P = 0.001), and 74, 141, 486, 490 bp T-RFs were all significantly lower than healthy controls (3.2% (1.3%, 5.1%) vs 10.2% (5.4%, 17.3%), P = 0.001; 4.5% (1.7%, 7.1%) vs 10.8% (5.9%, 21.1%), P = 0.007; 4.2% (1.6%, 5.3%) vs 7.6% (5.9%, 9.3%), P = 0.022; 3.6% (2.4%, 6.1%) vs 18.3% (9.9%, 43.2%), P = 0.008). The mucosal bacterial community composition in CD patients was predominated by Firmicutes, Proteobacterium and Actinobacterium. Compare with healthy control, Bacteroides were significantly reduced in CD patients while Firmicutes (e.g. Enterobacter sp.) and Actinomycetaceae significantly increased. CONCLUSIONS: Dysbiosis of mucosal microbiota occurs in CD with decreases of richness and biodiversity. Increased Enterobacter sp., Actinobacterium and decreased Bacteroides may play an important role in the pathogenesis of CD.

[A retrospective analysis of azathioprine in the treatment of 24 patients with refractory ulcerative colitis].

OBJECTIVE: To evaluate the efficacy and safety of azathioprine (AZA) in the treatment of refractory ulcerative colitis (UC). METHODS: Retrospective analysis of the clinical improvement, endoscopic improvement and mucosal healing rate, inflammation marker improvement after AZA administration and its safety in 24 refractory UC patients were performed, who were recruited between January 2007 and December 2011 in West China Hospital, Sichuan University, China. RESULTS: Twenty-four patients were enrolled, with a median age of 36 years old and a median course of 4 years. Among them, 14 cases were moderate UC and 10 cases were severe UC. The patients were treated with AZA in a dose of (1.23 ± 0.34) mg×kg(-1)×d(-1) from 7 weeks to 42 months. Efficacy was judged by Mayo disease activity index. At 3 months, 6 months and 1 year after treatment, the effective rates were 73.9% (17/23), 81.8% (18/22) and 14/16 respectively, and the remission rates were 17.4% (4/23), 54.5% (12/22) and 12/16 respectively. Both ESR and C reactive protein level after treatment for 6 months and 1 year were significantly lower than those before treatment [(9.3 ± 8.9) mm/1h, (10.9 ± 7.3) mm/1h vs (22.3 ± 10.7) mm/1h; 2.5(1.0-22.3) mg/L, 2.3(1.0-28.0) mg/L vs 18.4(3.6-137.0) mg/L; all P < 0.05]. Corticosteroid withdrawal rates at 3 months and 1 year after AZA treatment were 16/18 and 15/16, respectively. At 6 months and 1 year after AZA treatment, the endoscopic improvement rates were 85.7% (18/21) and 13/15 respectively; the endoscopic remission rates were 61.9% (13/21) and 11/15 respectively; and the mucosal healing rates were 61.9% (13/21) and 11/15 respectively. Adverse effects were occurred in 8 patients. Leukopenia was the most common adverse effect, followed by liver function injury, alopecia and epigastric discomfort. CONCLUSIONS: AZA is effective in the treatment of refractory UC patients with a low dose of (1.23 ± 0.34) mg×kg(-1)×d(-1), especially in the steroid withdrawing, maintaining remission and mucosal healing without severe adverse effects.

[Effects of ginsenoside Rg1 on expression of insulin-like growth factor-1 in brain of rats with brain contusion].

OBJECTIVE: To study the effects of ginsenoside Rg1 on the expression of insulin-like growth factor-1 (IGF-1) in the brain of rats after the experimental brain contusion. METHODS: A total of twenty-six Wistar rats were randomly divided into normal control group (n=2), untreated group (n=8) and ginsenoside Rg1 group (n=16). Brain injuries were induced in rats by a mechanical striking device. The brain tissues were extracted at different times after brain injury (6th hour, 12th hour, 2nd day, 6th day), then the expression of IGF-1 in brain tissue was examined by immunohistochemical method. RESULTS: In comparison with the normal control group, the expression of IGF-1 in the brain tissues was increased in the untreated group after the brain contusion (P<0.05). The expression of IGF-1 in brain tissues in ginsenoside Rg1 group was significantly increased as compared with the untreated group (P<0.05). CONCLUSION: Ginsenoside Rg1 enhances the recovery of the contused brain through increasing the expression of IGF-1.