Jieshou Li

Association of IL-1beta gene polymorphism with cachexia from locally advanced gastric cancer

BackgroundIL-1beta has been implicated in inflammatory episode. In view of the inflammatory nature of cancer cachexia, we determined the predictive value of IL-1B-31 T/C, -511 C/T, +3954 C/T and IL-1RN VNTR gene polymorphisms on the occurrence of cachexia associated with locally advanced gastric cancer.MethodsThe study included 214 patients and 230 healthy volunteers. Genomic DNA was prepared from peripheral blood leukocytes. Genotypes and allele frequencies were determined in patients and healthy controls using restriction fragment length polymorphism analysis of polymerase chain reaction products.ResultsThe overall frequencies of IL-1B-31 T, -511 T, +3954 T and IL-1RN VNTR alleles in patients with locally advanced gastric cancer were all comparable with those in controls. No significant differences were found in the distribution of IL-1B-31 T, -511 T and IL-1RN VNTR between patients with cachexia and without. Patients with cachexia showed a significantly higher prevalence of IL-1B+3954 T allele than those without (P = 0.018). In a logistic regression analysis adjusted for actual weight, carcinoma location and stage, the IL-1B+3954 CT genotype was associated with an odds ratio of 2.512 (95% CI, 1.180 – 5.347) for cachexia.ConclusionThe IL-1B+3954 T allele is a major risk for cachexia from locally gastric cancer. Genetic factors studied are not likely to play an important role in the determination of susceptibility to locally advanced gastric cancer.

Disruption of tight junctions during polymicrobial sepsis in vivo

The disruption of intestinal epithelial tight junctions may result in barrier function dysfunction during polymicrobial sepsis. The pathophysiology of sepsis involves breakdown of barrier integrity, which correlates with adverse outcome during sepsis. However, the mechanisms underlying loss of barrier function in sepsis remain unknown. In the present study in mice, tight junction (TJ) structure was analysed by transmission electron microscopy; intestinal permeability was assessed using molecular tracer measurement; and the distribution of TJ proteins was investigated by immunofluorescence microscopy. The membrane microdomains of TJs were isolated using discontinuous sucrose density gradients and the expression of TJ proteins in these was determined by western blot. Immunofluorescence microscopy revealed that claudins 1, 3, 4, 5, and 8 were present predominantly in the microvillous surface of epithelial cells and along the lateral membranes of the cells; in sepsis, however, labelling of these proteins was present diffusely within cells and was no longer focused at the lateral cell boundaries. Moreover, the expression of claudin‐2 was markedly up‐regulated in sepsis. Using western blot analysis, we found that occludin and claudins were displaced from raft fractions to non‐raft fractions in membrane microdomains of TJs in sepsis. In addition, the disruption of TJ structure was accompanied by increased intestinal permeability. Our results demonstrate for the first time that redistribution of TJ proteins in TJ membrane microdomains and redistribution of claudins in epithelial cells of the colon lead to alteration of TJ architecture and TJ barrier dysfunction during the development of polymicrobial sepsis. Copyright

Triptolide ameliorates IL-10-deficient mice colitis by mechanisms involving suppression of IL-6/STAT3 signaling pathway and down-regulation of IL-17

Triptolide is an active component of extracts derived from the medicinal vine Tripterygium Wilfordii Hook. f. (TWHF) whose extracts have been used to treat inflammatory bowel disease (IBD). We have reported that triptolide showed therapeutic activity in a murine IBD model, but the potential mechanism of action of this agent in IBD remains elusive. Accumulated data showed that both T-helper (Th) 1 and Th17 response may contribute to pathogenesis of human IBD and animal colitis. Interleukin (IL)-6/signal transducer and activator of transcription-3 (STAT3) signaling pathway play an important role in Th17 response as well as pathophysiology of IBD. We hypothesized that triptolide would attenuate the experimental colitis by repressing IL-17 and that this would involve down-regulation of IL-6/STAT3 signaling pathway. Histological examination demonstrated that triptolide significantly reduced the severity of colitis in C3H/HeJBir.IL-10-deficeint mice. Triptolide suppressed the IL-6/STAT3 signaling pathway, as well as repressed gene expression of IL-17 in vivo. In addition, triptolide (20ng/ml) in vitro was able to down-regulate the IL-6/STAT3 pathway and reduce IL-17 expression in cultured colonic explants from patients with Crohns disease (CD).

Epithelial-specific blockade of MyD88-dependent pathway causes spontaneous small intestinal inflammation

Accumulating evidence suggests a role for Toll-like receptor (TLR) signaling at the intestinal epithelial cells (IECs) level for intestinal protection against exogenous injury or pathogenic infection. We hypothesized that MyD88 dependent TLR signaling at intestinal epithelium is critical for mucosal immune homeostasis. In the current study, a transgenic mouse model was generated in which a dominant-negative mutant of MyD88 (dnMyD88) was driven by an intestinal epithelial-specific murine villin promoter. Aged transgenic mice spontaneously developed chronic small intestinal inflammation, as revealed by increased CD4+ and CD8+ lymphocytes, neutrophil and macrophage infiltration, increased production of cytokines as TNF-alpha, IFN-gamma, IL-1beta, and IL-17, crypt abscesses, lymphedema, and Goblet cell depletion. The chronic inflammation was not due to increased epithelial apoptosis or permeability, but to a decreased Paneth cell-derived alpha-defensins (cryptdins) and RegIII-gamma and increased commensal bacteria translocation. Thus, epithelial MyD88-dependent pathway plays an essential role in limiting mucosal microflora penetration and preventing mucosal immunoregulation disturbance in vivo.

Multimodal optimization of surgical care shows beneficial outcome in gastrectomy surgery.

BACKGROUND The aim of this trial was to compare multimodal optimization with conventional perioperative management in a consecutive series of patients undergoing gastrectomy procedures. METHODS According to randomized controlled studies and conclusions made by meta-analyses in colorectal surgery, optimized perioperative measures were designed and applied in gastrectomy surgery. Thirty-three patients were randomized to the optimized group and 30 patients to a control group. Two groups were treated in 1 center by a single surgical team in different wards. Both groups used patient-controlled intravenous analgesia for postoperative analgesia. The primary end point was length of postoperative hospital stay. Secondary outcomes included bowel function recovery after surgery, perioperative changes of inflammatory factors, glucocorticoid, insulin resistance, and body composition. Perioperative complications and adverse events were also recorded. RESULTS The groups were similar in terms of age, sex ratio, and Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity (POSSUM score). The optimized group was associated with a significantly shorter postoperative hospital stay compared with the conventional care group (P < .001). Durations of urinary catheterization and abdominal drainage were also less (P < .001). The diet program in the optimization group was well tolerated and was associated with an earlier recovery of gut function (P < .001). Proinflammatory factors were less elevated and body composition was more stable in the optimized group than in controls. There were no differences in morbidity or mortality between the groups. CONCLUSIONS Optimization of care in gastrectomy can shorten postoperative hospital stay and provides multiple beneficial outcomes, including hastening the return of gut function, without increasing morbidity.

Oral glutamine ameliorates chemotherapy-induced changes of intestinal permeability and does not interfere with the antitumor effect of chemotherapy in patients with breast cancer: a prospective randomized trial.

Aims and Background Sixty patients with breast cancer were randomly assigned to oral glutamine or placebo pre-neoadju-vant chemotherapy (CEF regimen). Methods and Study Design Oral glutamine supplementation was continued for at least 12 days. Patients kept a daily record of diarrhea and stomatitis. The plasma glutamine level, intestinal permeability (lactulose-mannitol test), and tumor size were analyzed. The expression of Ki-67 and PCNA antigens in breast carcinoma was assessed. Results The plasma glutamine level was significantly higher in the glutamine group than in the placebo group (420.39 ± 52.39 mmol/L vs 309.76 ± 42.34 mmoi/L, P <0.05). After one cycle of chemotherapy, the lactulose-mannitol ratio was higher in the placebo group than in the glutamine group (0.0630 ± 0.0091 vs 0.0471 ± 0.0094, P <0.05). No differences were observed in the grades of stomatitis and diarrhea, in the changes in tumor size, and in the expression of Ki-67 and PCNA antigens between the two groups. Conclusions Prophylactic oral glutamine could ameliorate the neoadjuvant chemotherapy-induced increase in intestinal permeability, but had no significant positive clinical effect on stomatitis and diarrhea and did not interfere with the antitumor effect of chemotherapy.

Association of two polymorphisms of tumor necrosis factor gene with acute severe pancreatitis

BACKGROUND Acute severe pancreatitis (ASP) has an unpredictable course and outcome. Polymorphisms of the tumor necrosis factor (TNF) gene have been related to TNF production and outcome in a variety of inflammatory diseases. We assessed whether TNF gene polymorphism and plasma concentrations of TNF-alpha were related to the occurrence of ASP and septic shock from it. METHODS The study included 208 patients (102 with ASP, 106 with acute mild pancreatitis) and 116 healthy volunteers. TNF-beta and TNF-alpha-308 polymorphisms were performed by analyzing NcoI-digested DNA fragment obtained by polymerase chain reaction (PCR). HLA-DRB1*0301 (HLA-DR3) typing was performed by sequence-specific oligonucleotide typing on PCR-amplified genomic DNA. Plasma concentrations of TNF-alpha were measured by solid-phase enzyme-amplified sensitivity immunoassay. RESULTS The overall TNF2 allele frequency and TNFB2 allele frequency were similar in patients with mild or severe pancreatitis. Further, no significant difference in gene polymorphism frequencies was noted between patients with acute pancreatitis and controls. TNF2 frequency was significantly increased in septic shock patients compared with nonseptic shock patients (53.8 vs 22.4%; P = 0.003), as was TNFB2 frequency (88.5 vs 63.2%; P = 0.015). A linkage disequilibrium between TNF2 allele and HLA-DR3 was seen, this haplotype was not associated with severity of acute pancreatitis or outcome of septic shock in ASP patients. With regard to the mortality rate in patients who developed septic shock, those with a TNF2 allele had a significantly higher rate of mortality than those with a TNF1 allele (expired: 8 vs 1, P = 0.018). Plasma levels of TNF-alpha did not differ significantly in ASP patients displaying different alleles of the TNF gene studied. CONCLUSIONS Results indicate that the TNF gene polymorphisms studied play no part in determination of disease severity or ASP susceptibility; however, they are both strongly related to the development of septic shock in ASP. TNF2 allele was associated with death as a result of ASP-associated septic shock.

Concomitant upregulation of nuclear factor-kB activity, proinflammatory cytokines and ICAM-1 in the injured brain after cortical contusion trauma in a rat model

BACKGROUND Nuclear factor kappa B (NF-kB), proinflammatory cytokines and intercellular adhesion molecule 1 (ICAM-1) are frequently upregulated in the injured brain after traumatic brain injury (TBI). However, the temporal pattern of upregulation is not well defined. AIMS The current study was undertaken to investigate the temporal profile of the expression of NF-kB, proinflammatory cytokines and ICAM-1 in the injured brain after cortical contusion trauma of the rat brain. SETTINGS AND DESIGN A rat model of cortical contusion was produced by a free-falling weight on the exposed dura of right parietal lobe. The rats were randomly divided into control group and TBI groups at hours 3, 12, 24 and 72, and on day 7. MATERIAL AND METHODS NF-kB binding activity in the surrounding brain of injured area was studied by electrophoretic mobility shift assay (EMSA). The levels of TNF-alpha and IL-6 were detected using ELISA and ICAM-1 expression studied by immunohistochemistry. STATISTICAL ANALYSIS The data were analyzed by one-way ANOVA followed by Student-Newman-Keuls post hoc test. Relation between variables was analyzed using bivariate correlation with two-tailed test. RESULTS Compared with that of control group, NF-kB binding activity in the injured brain was significantly increased through 12 h and 7 days postinjury, with the maximum at 72 h. The concentrations of TNF-alpha and IL-6 in the injured brain were significantly increased from 3 h to 7 days and maximal at 24 h postinjury. The number of ICAM-1 immunostained microvessels was significantly increased in the injured brain from 24 h to 7 days postinjury, with its peak at 72 h. Concomitant upregulation of TNF-alpha, IL-6, ICAM-1 and the cytokine mediators NF-kB in the injured brain was observed in the injured brain after cortical contusion, and there was a highly positive relation among these variables. CONCLUSIONS Cortical contusion trauma could induce a concomitant and persistent upregulation of NF-kB binding activity, TNF-alpha, IL-6 and ICAM-1 in the injured rat brain which might play a central role in the injury-induced immune response of brain.

Triptolide ameliorates Crohn's colitis is associated with inhibition of TLRs/NF-κB signaling pathway.

Growing evidence suggests that TLRs/NF-κB signaling pathway plays a critical role in the pathogenesis of Crohns disease (CD). We have reported that triptolide, an active component from Tripterygium wilfordii Hook, showed therapeutic activity in IL-10-deficeint (IL-10-/- mice, a murine CD model. However the full mechanisms of action of this agent in CD remain largely unknown. We hypothesized that triptolide would ameliorate the experimental colitis by inhibiting TLRs/NF-κB signaling pathway. We found TLR2 and TLR4 were upregulated in IL-10-)/- mice, triptolide inhibited the TLRs/NF-κB signaling pathway in vivo. In addition, triptolide in vitro was able to downregulate the TLRs/NF-κB pathway in cultured colonic explants from CD patients. Our results confirm the therapeutic effect of triptolide in experimental colitis, and suggest it as a promising compound for CD treatment. These findings also support the possibility that targeted inhibition of TLR signaling pathway is an approach deserving further investigation as a therapeutic strategy for CD.

Fish Oil Enhances Recovery of Intestinal Microbiota and Epithelial Integrity in Chronic Rejection of Intestinal Transplant

Background The intestinal chronic rejection (CR) is the major limitation to long-term survival of transplanted organs. This study aimed to investigate the interaction between intestinal microbiota and epithelial integrity in chronic rejection of intestinal transplantation, and to find out whether fish oil enhances recovery of intestinal microbiota and epithelial integrity. Methods/Principal Findings The luminal and mucosal microbiota composition of CR rats were characterized by DGGE analysis at 190 days after intestinal transplant. The specific bacterial species were determined by sequence analysis. Furthermore, changes in the localization of intestinal TJ proteins were examined by immunofluorescent staining. PCR-DGGE analysis revealed that gut microbiota in CR rats had a shift towards Escherichia coli, Bacteroides spp and Clostridium spp and a decrease in the abundance of Lactobacillales bacteria in the intestines. Fish oil supplementation could enhance the recovery of gut microbiota, showing a significant decrease of gut bacterial proportions of E. coli and Bacteroides spp and an increase of Lactobacillales spp. In addition, CR rats showed pronounced alteration of tight junction, depicted by marked changes in epithelial cell ultrastructure and redistribution of occuldin and claudins as well as disruption in TJ barrier function. Fish oil administration ameliorated disruption of epithelial integrity in CR, which was associated with an improvement of the mucosal structure leading to improved tight junctions. Conclusions/Significance Our study have presented novel evidence that fish oil is involved in the maintenance of epithelial TJ integrity and recovery of gut microbiota, which may have therapeutic potential against CR in intestinal transplantation.