Objective To investigate the effect and molecular mechanism of isolongifolene(ISO)on the apoptosis of intestinal epithelial cells and 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced Crohn's disease(CD)-like colitis in mice.Methods In the animal experiments,mice were randomly assigned to the wild type(WT)group,TNBS group and TNBS+ISO group,with 8 mice in each group.Colitis models of mice were established in the TNBS group and the TNBS+ISO group by rectal injection of TNBS.After modeling,the mice in the TNBS+ISO group were given ISO intervention via intragastric gavage(10 mg/kg),and the other two groups were given the same amount of normal saline via intragastric gavage.The mice were sacrificed on the 7th day.The changes in body mass,disease activity scores(DAI),and the colon length of mice were measured,and transepithelial electrical resistance(TEER)of the colon tissues was determined.The score of colon inflammation was calculated according to HE staining.The levels of intestinal mucosal inflammatory factors,including tumor necrosis factor alpha(TNF-α),interferon(IFN)-γ,interleukin(IL)-1 β,and IL-6,were measured by RT-PCR and ELISA.The apoptosis of colon tissue cells was determined by TUNEL assay.The expressions of apoptotic proteins(cleaved caspase-3/caspase-3 and Bax),an anti-apoptotic protein(Bcl-2),and tight junction proteins(ZO-1 and claudin-1)were detected by Western blot and immunofluorescence.In the cell experiment,TNF-α was used to induce intestinal epithelial cell Caco-2 apoptosis model,which was treated with ISO.Then,intervention with the AMPK inhibitor Compound C was given.TUNEL assay,Western blot assay,and immunofluorescence assay were performed to measure apoptosis and the expression of apoptosis proteins in the Caco-2 cells.Gene Ontology(GO)enrichment analysis was performed to predict the biological function of ISO.Then,the mechanism involved was verified by examination of the mice and Caco-2 cells.Western blot was performed to determine the expression levels of p-AMPK/AMPK and p-PGC1α in the colon tissues from the mice of different groups and Caco-2 cells.The apoptosis of the cells was determined by TUNEL assay.Results According to the results of the animal experiment,ISO could alleviate experimental colitis and intestinal barrier dysfunction,leading to improvements in body mass loss,colon length shortening,DAI score,inflammatory rating,and TEER values(all P＜0.05)in mice.Furthermore,ISO decreased the expression of pro-inflammatory factors TNF-α,IFN-γ,IL-1β,and IL-6 and increased the expression of the tight junction proteins ZO-1 and claudin-1(all P＜0.05).In the cell experiment,in a TNF-α-induced intestinal epithelial cell model,ISO was also found to protect intestinal barrier against damage.ISO reduced the proportion of apoptotic intestinal epithelial cells,reduced the expression of cleaved-caspase-3/caspase-3 and Bax,and upregulated the level of Bcl-2(all P＜0.05).GO enrichment predictive analysis showed that the role of ISO in improving CD-like enteritis might be associated with the negative regulation of apoptosis.Verification of the mechanism showed that the expression of p-AMPK and p-PGC1α in the mice colon tissue was significantly upregulated after ISO intervention(P＜0.05).In contrast,the AMPK inhibitor Compound C increased the apoptosis rate of ISO-treated Caco-2 cells and decreased the relative expression levels of ZO-1 and claudin-1(P＜0.05).Conclusion ISO reduces intestinal epithelial cell apoptosis at least in part by activating AMPK/PGC1α signaling pathway,thereby alleviating TNBS-induced intestinal barrier dysfunction and CD-like colitis in mice.

Objective To observe the clinical effects of tibial locking multidirectional interlocking intramedullary nail (TLMIIN) for tibial plateau fractures. Methods 38 cases with closed tibial plateau fractures treated by TLMIIN from October 2008 to May 2012 were retrospectively analyzed. There were 22 males and 16 females, with an average age of 48.7 years (range, 28-67 years). There were 24 fractures on the left side while the other 14 factures were on the right side, which were all fresh frac?tures. According to AO/OTA classification of tibial plateau fractures, there were 4 cases of B1, 1 case of B2, 14 cases of B3, 8 cas?es of C1, 5 cases of C2 and 6 cases of C3. Close reduction were performed on 4 cases (10.5%of all cases). Open reduction were performed on the other 34 cases. The limited incision was decided by the distribution of the fragments and fracture line. Hohl?Luck evaluation system was applied for the follow?up. Results The mean follow?up period was 18.1 months (range, 11-23 months). All fractures were healed at an average period of 87.4 days (range, 48-131 days). The average time from operation to full weight?bearing was 108.9 days (range, 80-128 days). Hohl?Luck evaluation system was used in the final follow?up. The excellent and good rate of functional score was 94.7%(36/38), including 28 cases excellent, 8 cases good and 2 cases fair from functional as?pect. The excellent and good rate of radiological score was 84.2%(32/38), including 23 cases excellent, 9 cases good and 6 cases fair from radiological aspect. No complications such as infection, breakage and loosening of the screw, malunion, nonunion oc?curred at the time of the latest follow?up. 6 cases with serious swelling of the knee joint and the soft tissue of crus were cured by an?ticoagulation, dehydration and physiotherapy treatments after operation. The 2 cases with a little exudates and incrustation was bacterial cultured negative and healed after 16 days and 18 days on the incision. 1 case had lost of reduction due to weight?bearing 1 week after operation, who had 25 degree of varus deformity, which was left dispose, and the bone healing and joint function were unaffected. Conclusion The intramedullary support, restrictive and non?restrictive multidirectional tridimensional fixation of TLMIIN technology had satisfactory effects in treating plateau fractures. It can supply a new therapeutic method, with little dam?age of soft tissues when taking out the internal fixation and reduce some postoperative complications.