Objective To investigate the effects of lymph from ischemic/reperfused intestine on the inflammatory factors and Toll-like receptor 4 (TLR4) ligand high mobility group box-1 (HMGB1) in TLR4 deficient (TLR4-/-) mice.Methods A total of 20 SD rats weighing (300 ±20) g were randomly assigned into two groups:lymph drainage group (group N,lymph drainage for 180 minutes without other treatment) and intestinal ischemia/reperfusion group (group I/R,draining the lymph for 180 minutes while clipping the superiormesenteric artery for 60 minutes followed by 120-minute reperfusion).Thirty-two TLR4-/-mice and thirty-two C57BL/6 wild type (WT) mice were each divided into 4 sub-groups (n =8),injected with different fluids through the caudal vein:group N with normal lymph;group I/R with I/R lymph;group Edt with endotoxin;group HMGB1 with HMGB1 protein.The mice were sacrificed 180 minutes after the injection for sample collection.Results The levels of endotoxin and HMGB1 in the lymph drainage of the group I/R rats were significantly higher than that of the group N rats [(0.034 ± 0.050) Eu/ml vs.(0.017 ± 0.023) Eu/ml,P =0.033;(4.293 ± 0.883) ng/ml vs.(0.509 ± 0.128) ng/ml,P =0.006].In the mice injected with HMGB1,the mucosa thickness and villus height in the ileum of the WT mice were significantly lower than that of the TLR4-/-mice [(335.8±43.2) μmvs.(602.1±37.5) μm,P=0.000;(273.0±31.7) μm vs.(404.5 ± 18.6) μm,P =0.000];in both WT and TLR4-/-mice injected with the I/R lymph drainage,the mucosa thickness and virus height were decreased,but the decrements were significantly lower in TLR4-/-mice;there were no statistically significant differences in the levels of interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),endotoxin,and HMGB1 between the TLR4-/-and the WT mice injected with normal lymph or endotoxin.In the mice injected with I/R lymph drainage,the levels of inflammatory factors in the TLR4-/-mice were significantly lower than those in the WT mice [TNF-α:(28.637 ±5.166) pg/ml vs.(41.917 ±8.175) pg/ml,P=0.000;IL-6:(60.900 ±24.729) pg/ml vs.(110.265 ±28.545) pg/ml,P =0.000].In the mice injected with HMGB1,the levels of inflammatory factors in the TLR4-/-mice were significantly decreased compared with those in the WT mice [TNF-α:(20.865 ± 6.464) pg/ml vs.(31.059 ± 6.204) pg/ml,P=0.004;IL-6:(36.268 ±8.977) pg/ml vs.(76.677 ± 14.099) pg/ml,P=0.000].Conclusions The concentrations of endotoxin and HMGB1 are significantly increased during intestinal I/R in rats.After injection of I/R lymph drainage,endotoxin,and HMGB1,the levels of inflammatory factors and HMGB1 in the mice injected with I/R lymph drainage are significantly higher than those in the mice injected with normal lymph;the levels of inflammatory factors and local damage of intestinal mucosa are significantly reduced in the TLR4-/-mice than in the WT mice.The gut-lymph pathway may play a key role in the intestinal I/R injury.

Objective To investigate the effects of arsenic trioxide (ATO) on the expression of autoan-tibody and interleukin (IL)-10 IL-12 in MRL/lpr mice. Methods MRL/lpr mice wereseparated into 3 different groups. The 3 groups received arsenic trioxide (ATO, 0.4 mg·kg~(-1)·d~(-1)), cyclophosphamide (CTX,50 mg/kg) and sodium chloride (NS, volume weight-determined) abdominal injection respee-tively. The treatment stopped 2 months later. Afterwards, the rates of CD3~+(T) cells, CD3~+CD4~+(Th) cells and the CD3~+CD4~+cells which produced IL-10 and IL-12 were detected using single-cell measurement of intr-acellular eytokines by flow cytometry after polyclonal stimulation with PMA and ionomycin for 4 hours in 5% CO_(2.)Serum levels of IL 10 and IL-12 were assessed using the Mouse cytokines ELISA Kit. One-way ANOVA LSD test and paires t test were used for statistical analysis.Results ①The level of anti-dsDNA antibody after treatment was 0.92±0.06, while it was 1.14±0.58 before treatment. So the ds-DNA antibody level was significantly decreased in ATO group (P<0.01), while it was dramatically increased in the NS groups (P<0.05) after the treatment;②ATO group had significantly less CD3~+ cells and CD3~+CD4~+ cells[(44±4)% and (20±4)%]compared withNS group [(59±5)%and(30±3)%](P<0.01).③The serum level of IL-12 in the ATO group was (84±12) pg/ml,while it was (103±13)pg/ml in the NS group (P=0.018).④The intracellular levels of IL-10 and IL-12 produced by CD3~+CD4~+ (Th) cells in the ATO group were ( 1.5±0.4)% and (2.43±0.42)%, which was significantly lower than those in the NS group respectively (2.5±0.5)% and (3.24±0.40)%(P<0.01). Conclusion Arsenic trioxide can reduce the production of anti-dsDNA antibody,inhibit the activation and proliferation of both T cells and Th subsets in the MRLApr mice, and hence decrease the serum levels of IL-12 and the levels of IL-10, IL-12 produced by Th cells.

Objective To explore the expressions and correlation of RWD containing sumoylation enhancer (RSUME),small ubiquitin-like modifier (SUMO-1),hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor (VEGF)in gliomas of different pathologic grade.Methods We investigated the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA with reverse transcription-polymerase chain reaction (RT-PCR),and investigated the immunohistochemical staining to determine the expressions of SUMO-1,HIF-1α and VEGF in 63 cases of human gliomas of different pathologic grade and 9 cases of normal brain tissues.We studied its correlation with the pathologic grade and the relationship between the expression of RSUME promoter sumoylation and HIF-1α/VEGF pathway in gliomas.Results There were significant differences (P <0.01)in the expressions of RSUME mRNA,HIF-1αmRNA and VEGF mRNA in glioma tissues.With the increasing degree of pathologic grade in tumor specimens,the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA increased markedly (P <0.01 ).There was a positive correlation of the expression levels of RSUME mRNA with HIF-1αmRNA and VEGF mRNA.There were significant differences (P <0.01 )in the expressions of SUMO-1,HIF-1αand VEGF in glioma tissues by immunohistochemical staining.With the ascending of pathologic grade of tumor specimens,the expression levels of SUMO-1,HIF-1α and VEGF increased markedly (P < 0.01 ).There was a positive correlation between the expression level of SUMO-1 and HIF-1α(r =0.857,P <0.01).The Kaplan-Meier analysis and Log-rank test showed significant differences in progress free survival (PFS)between the RSUME high-expression and low-expression groups (χ2 =36.032,P <0.01).Conclusion RSUME may enhance HIF-1α/VEGF pathway through sumoylation in gliomas.It implicates that RSUME is related to angiogenesis in gliomas and can promote tumor invasion and progression,indicating that RSUME can be a novel target in gliomas treatment.

AIM:To study the effect of centromere protein W ( CENP-W) down-regulation on human glioma U87 cells.METHODS:Small interfering RNA ( siRNA) was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot .The proliferation of the cells was analyzed by MTT assay , BrdU staining and colony formation experiment .Transwell chamber assay was used to detect the invasion a-bility of the cells .The cell migration ability was measured by a scratch test .The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry .RESULTS:The results of MTT assay , colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group .The results of Transwell and scratch experiments showed that the migra-tion and invasion abilities in experimental group were weaker than those in blank control group and negative control group . The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G 0/G1 phase .The percentage of apoptotic cells in experimental group was higher than that in control group ( P<0.05 ) .CON-CLUSION:Down-regulation of CENP-W expression inhibits the proliferation , migration and invasion of human glioma cells and promotes the apoptosis of the cells , suggesting that CENP-W may be a potential target of gene therapy for human glioma.

Objective To investigate the expression of CENP-W in gliomas and its relationship with clinicopathological parameters and prognosis and to explore the effects of centromere protein W (CENP-W)on the invasion of gliomas cells.Methods The expressions of CENP-W in high-grade glioma tissues,low-grade glioma tissues,and adjacent brain tissues were detected by real-time fluorescence quantitative PCR and Western blotting. The correlation of the expression of CENP-W with clinicopathological parameters and prognosis was analyzed statistically.Human gliomas U2 5 1 cells in vitro were transfected with small interfering RNA to downregulate the expression of CENP-W.The invasion and migration capabilities of gliomas cancer cells were assessed by Transwell assays.Results The expression level of CENP-W was significantly higher in glioma tissues than in normal tissues. There was a positive correlation between the three protein expression levels and the pathological grade of gliomas. CENP-W siRNA was successfully transfected into U2 5 1 cells.Compared with those of the cells transfected with the scramble siRNA and control cells,the invasive and migration activities were inhibited in the U2 5 1 cells transfected with CENP-W siRNA.The Kaplan-Meier analysis and Log-Rank test showed significant differences in progress free survival (PFS)between the CENP-W high-expression and low-expression groups.Conclusion The expression level of CENP-W was positively correlated with the pathological grade of gliomas and CENP-W can promote glioma cell invasion.It implicates that CENP-W can be a novel target in gliomas treatment.

Objective To evaluate the effects of ambroxol on prevention of bronchopulmonary complications after thoracotomy in patients with chronic obstructive pulmonary disease(COPD).MethodsTotal 161 elderly patients aged ≥70 years with COPD undergoing thoracotomy were randomly allocated to two groups:ambroxol group (300 mg/d,6 d) and placebo group as control.Pulmonary complications were evaluated by clinical symptoms,radiographic changes,and blood gas analysis. Results The incidence rates of atelectasis were 8.6％ in ambroxol group and 28.8％ in placebo group,respectively.The PaO2 values after surgery in ambroxol group decreased more than in placebo group (P＜0.05) compared with the preoperative values.Side effects were not found in all patients.Conclusions Ambroxol should be considered as an alternative pharmacologic approach for the prevention of post-thoracotomy pulmonary complications in the elderly patients with COPD.

ObjectiveBronchogenic carcinoma involving the carina or tracheobronchial angle still presents a challenge due to specific problems related to surgical technique and airway management.Aim of this paper is to examine complications and long-term survival of our personal series and those reported in literature.MethodsBetween 1985 and 2010,48 patients underwent carinal resection:a right tracheal sleeve pneumoneetomy was performed in 47 patients and a left tracheal sleeve pneumonectomy in 1 patient.The anastomosis was performed with aid of high-frequency jet ventilation or introfield tube ventilation.ResultsOverall morbidity and mortality rate was 25％ and 6.3％ respectively,and there was no death in operation.5-yearsurvival rate of patients with squamous and adenocarcinoma was 27.3％ and 12.5％,respectively,P =0.04.The overall 5-year survival rate was 24.3％.Patients without nodal involvement had a significantly better prognosis than N1 and N2 patients (5-year survival:52％,13％ and 0,respectively).Multivariate analysis showed that nodal status was the only independent prognostic factor( P =0.006 ).ConclusionWith careful selection of patients and meticulous surgical technique,Tracheal sleeve pneumonectomy for bronchogenic carcinoma can be accomplished with acceptable mortality and morbidity,proriding good long-term results.

Objective To evaluate the complications and prognostic factors of lung transplantation performed in a single center.Methods A rettospective analysis of demographic and outcome data of lung transplantation was performed.Survival analyses were performed using Kaplan-Meier estimation.Results Between January 2003 and April 2011,42 lung transplant procedures were performed.Overall survival rate at 1,3,and 5 years were 89％,59％ and 38％,respectively.1,3,and 5 years survival in patients with COPD was 83％,66％ and 45％,respectively,which were better than other primary end stage lung diseases ( 78％,17％ and 17％,respectively,P =0.013).Postoperative complications included pulmonary bacterium infection in 8 patients (20％),fungal infection in 12 (30％),and airway complications in4 (9.5％).35％ of patients had at least 1 episode of acute rejections within the first year,and 22.5％ of patients had BOS.2 patients underwent single lung retransplantation.Conclusion In this single center study,patients with COPD may have a good long-term survival.The most common postoperative complications were pulmonary infection and airway complication.

Objective To investigate the effect of different antibodies on Toll-like Receptor 4-High Mobility Group Box 1 and its downstream signal transductions in distant organ injuries caused by intestinal ischemia/reperfusion in mice.Methods A total of 40 mice (C57BL/6,SPF level) were by random number table method assigned into five groups:sham,control,anti-HMGB1,anti-Myeloid differentitation gene,and antiTIR domain containing adaptor inducing IFN-β (n=8).In the control,anti-HMGB1,anti-MyD88,and antiTRIF groups,the IgG,HMGB1,MyD88,and TRIF antibodies were injected,respectively,via the tail vein 30 minutes before ischemia (1 mg/kg body weight,0.025％).After anesthesia and abdomen incision,all mice,except the sham group,underwent intestinal ischemia by clamping the superior mesenteric artery for 60 minutes followed by 60 minutes of reperfusion.Sham group underwent the same surgical procedures except for clamping the artery.Serum nuclear factor-κB p65,Interleukin-6 and Tumor Necrosis Factor-α were measured.Morphological changes in the lung and intestine were evaluated.mRNA and protein expressions of HMGB1 and NF-κB in lung and intestinal tissues were assayed.Results Compared with the control group [(228.53± 24.85),(104.91±31.18),and (70.81±46.97) ng/L],HMGB1 [(145.00±33.63),(62.28±6.73),and (52.76± 5.71) ng/L],MyD88 [(191.12± 13.22),(85.90± 17.37),and (63.19 ± 5.47) ng/L],and TRIF [(183.73±10.81),(78.14±7.38),and (59.70±4.63) ng/L] significantly decreased the serum level of NF-κB (P=0.000,0.005,0.001),IL-6 (P=0.000,0.004,0.000) and TNF-α (P=0.000,0.024,0.002) after ischemia reperfusion.Tissue injuries in the lung and intestine were also alleviated by HMGB1,MyD88,and TRIF.The anti-HMGB1,anti-MyD88,and anti-TRIF groups displayed significant elevations of HMGB1 mRNA [lung (1.89±0.18),(2.35±0.31),and (2.29±0.28),ileum (4.93±0.55),(5.96± 0.73),and (5.76±0.51)],NF-κB mRNA [lung (1.42±0.23),(1.77±0.18) and (1.70±0.13),ileum (2.23±0.55),(3.11±0.38) and (2.99±0.24)] and NF-κB protein expressions in lung and ileum tissues compared to the sham group [lung HMGB1 mRNA (1.04±0.19) (P=0.000,0.000,0.000),NF-κBmRNA (1.03±0.21) (P=0.004,0.000,0.000),ileum HMGB1 mRNA (1.14±0.54) (P=0.000,0.000,0.000),NF-κB mRNA (1.03±0.23) (P=0.000,0.000,0.000)].However,incornparison with the control group [lung HMGB1 mRNA (2.67±0.23) (P=0.000,0.035,0.016),NF-κB mRNA (2.04±0.29) (P=0.000,0.039,0.012),ileum HMGB1 mRNA (6.70±0.66) (P=0.001,0.038,0.015),NF-κBmRNA (3.71±0.53) (P=0.000,0.018,0.006)],the other three groups showed a significant down-regulation,with the most remarkable decrement in the anti-HMGB1 group.Application of anti-HMGB1,anti-MyD88,and anti-TRIF could drastically attenuate the tissue injuries in ischemia reperfusion.anti-HMGB1 exhibited the most significant effect.Conclusions HMGB1 and its downstream signals play an important role in intestinal ischemia reperfusion injuries in mice.Of two downstream signals,the TRIF-dependent pathway exerts a more important effect than that of the MyD88-dependent pathway.

Objective To investigate the clinical and prognostic significance of ABO promotor methylation level in adult patients with leukemia and myelodydysplastic syndrome(MDS). Methods ABO promoter methylation level of 182 malignant hematological disease patients and 68 normal controls were detected by bisulfite sequencing PCR. Then clinical features and outcome were compared between hypermethylation group and hypomethylation group. Results The median methylation rate of ABO promoter in newly diagnosed acute myeloid leukemia (AML) and acute lymphocytic leukemia (ALL) were 46.98％ and 11.01％ respectively, which were both higher than that in controls (2.30％, P<0.05). The methylation rates in remission AML and ALL were 1.58％and 2.30％respectively, which were comparable with that in normal group (P>0.05). As to relapse AML and ALL, methylation rates were 41.26％ and 17.50％respectively, also significantly higher than that in controls (P<0.05).In patients with chronic myeloid leukemia (CML) chronic phase, the median methylation rate was 1.00％, which was similar to normal group. But a CML patient who transformed to ALL hadextremely high methylation rate 92.56％. The median methylation rate in patients with MDS significantly elevated as 5.81％ compared with that in controls (P<0.05). The median overall survival (OS) of ALL and AML (non-M3) patients with hypermethylation were 12.5 months and 15.3 months, which were significantly shorter than those with hypomethylation (24.0 months and 20.0 months) (P<0.05). The median disease-free survival (DFS) of ALL and AML (non-M3) patients with hypermethylation were 9.9 months and 12.0 months, which were significantly shorter than those with hypomethylation (22.3 months and 18.5 months), (P<0.05). Multivariable analysis suggested that ABO promoter methylation level was an independent predictive factor of OS and DFS in ALL and AML (non-M3) patients. Conclusion ABO promoter hypermethylation is closely related to genesis, development and prognosis of leukemia and MDS. Hypermethylationis related to a clinical poor prognosis compare with hypomethylation.