Objective To study the influence on heat shock protein 27 (Hsp27) and tumor necrosis factor ? (TNF-?) by inhibiting ubiquitin proteasome system in cardiac ischemia-reperfusion (I/R) rats. Methods Left anterior descending (LAD) coronary artery was ligated for 30 min and then released for 6 h in adult Sprague-Dawley rats. Five minutes before reperfusion, MG-132 (N-benzoyloxycarbonyl (Z)-Leu-Leuleucinal) at dose of 0.75 mg/kg was given by vein injection in I/R+T group, but I/R group, I group and sham group were given the same volume of sterile saline. The changes of myocardial infarct size, cardiac protein and mRNA levels of Hsp27 and TNF-? were measured. Results Compared with I/R group, the myocardial infarct size was significantly reduced (P

Objective To explore the effects of rosiglitazone (ROSI),a peroxisome proliferator-activated receptors-gamma (PPAR-γ) ligand,on hyperlipidemia in rats with severe acute pancreatitis (SAP) associated with lung injury.Methods A total of 120 male SD rats received intragastric administration of high fat diet for two weeks to induce experimental hyperlipemia.The hyperlipidemic rats were randomly (random number) divided into six groups:hyperlipidemia (HL) group (n =20),hyperlipidemia with SAP (HP) group (n =20),hyperlipidemia with rosiglitazone intervention (HRP) group (n =20),hyperlipidemia with rosiglitazone and antagonist to rosiglitazone (HRGP) group (n =20),rosiglitazone control (HR) group (n =20) and antagonist control (HG) group (n =20).The SAP was induced by a retrograde infusion of 5％ sodium tauroholate into bile-pancreatic duct,and the SAP was established in HP group,HRP group and HRGP group.In HL group,HR group and HG group,equivalent volume of normal saline was used instead of sodium taurocholate.In HRP group and HR group,ROSI (6 mg/kg) was administered via the femoral vein 1 hour prior to the administration of sodium taurocholate.In HRGP group,GW9662 (0.3 mg/kg),an antagonist to PPRA-gamma,was given via the femoral vein 30 min prior to the administration of ROSI.In HG group,only GW9662 (0.3 mg/kg) was given via the left femoral vein 30 min prior to pretend SAP modeling.Rats from each group were sacrificed by exsanguination 12 h after SAP modeling.Blood samples were taken from all subjects to measure serum amylase (AMY),total cholesterol (TC),triglycerides (TG),Successive sections of the paraffin embedded tissue from pancreas and lung were taken for pathological examination with hematoxylin-eosin (HE) staining.Histopathological changes of pancreatic and pulmonary tissues observed under light microscope were evaluated.In pulmonary tissue,nuclear factor-kappa B (NF-κB) p65 expression was assayed by immunohistochemistry.Intercellular adhesion molecule (ICAM-1) protein and tumor necrosis factor-α (TNF-γ) protein levels were studied using Western blot analysis.Results The serum levels of TC and TG in HL group and HP group were significantly higher than those in HR group and HRP group (1.24 ± 0.28,1.14 0.08 vs.0.41 ±0.17,0.58±0.12;14.86±1.47,12.42±0.96 vs.6.52±2.04,7.36±0.95,allP＜ 0.05);The levels of serum AMY,W/D ratio,pancreas pathologic score,lung pathologic score,expression of NF-κB p65,ICAM-1 and TNF-α in pancreas in the HP group and HRGP group were significantly higher than those in HL group,HR group,and HG group (6 501.9 ±3 770.0,5 922.2 ±925.9 vs.1 139.3 ± 35.6,1 070.8 ±67.0,1 012.4 ±94.7;3.14±0.16,3.06±0.12vs.1.81 ±0.13,1.76±0.23,1.83 ± 0.18;all P ＜0.05);Compared with the HP group and HRGP group,the levels of serum AMY,TC and TG were significantly decreased in HR group and HRP group,ameliorating pancreas and lung pathological damage,and down-regulating the expression of NF-κB p65,ICAM-1 and TNF-α in pulmonary tissue (all P ＜ 0.05).While there were no statistically significant differences in above biomarkers between HP group and HRGP group (all P ＞ 0.05).Conclusions Our study demonstrates that ROSI exerts anti-hyperlipidemic effect and anti-inflammatory effect on hyperlipidemia in rats with sodium taurocholate-induced severe acute pancreatitis associated with lung injury by inhibiting NF-κB and down-regulating the expression of TNF-α and ICAM-1.

Objective To investigate the effects and mechanisms of poly adenosine diphosphate (ADP)-ribose polymerase inhibitor 3-aminobenzamide (3-AB) on kidney injury in rates with severe acute pancreatitis (SAP).Methods Fifty-six male Wistar rats were divided into the sham operation (SO) group,SAP (3,6,12 hours) groups,and 3-AB + SAP (3,6,12 hours) groups,and there were 8 rats in each group.SAP model was established by retrograde injection of 5％ sodium taurocholate into the biliopancreatic duct.Rats in the 3-AB + SAP group were infused with 3-AB (20 μg/g) via femoral vein 30 minutes before SAP model establishment.The serum amylase,kidney function and renal myeloperoxidase (MPO) were determined,and pathological scores of pancreatic and renal tissues were evaluated under light microscope.Renal poly ADP-ribose formation,intercellular adhesion molecules-1 (ICAM-1) and P-selectin expression were detected by the Western blot.All data were analyzed using the analysis of variance or t test.Renal injury grading was analyzed using the Kruskal-Wallis nonparametric test.Results The levels of serum amylase of SAP 3,6,12 groups were (3806 ± 229)U/L,(4898 ± 295) U/L and (5726 ± 372) U/L,which were significantly higher than (2785 ± 160) U/L,(3241 ± 198) U/L and (3953 ± 249) U/L of the 3-AB + SAP groups (t =3.652,4.672,4.407,P ＜ 0.05).The levels of blood urea nitrogen were (11.6 ± 0.8) mmol/L,(19.3 ± 1.3) mmol/L and (29.6 ± 2.1) mmol/L,which were higher than (7.5 ± 0.5) mmol/L,(10.5 ± 0.7) mmol/L and (21.6 ± 1.5) mmol/L of the 3-AB + SAP groups.There were significant differences in the levels of blood urea nitrogen between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.836,6.849,P ＜0.05).The levels of creatinine of the SAP 3,6,12 hours groups were (48.7 ±3.1) μmol/L,(58.3 ±3.7) μmol/L and (75.9 ±5.4) μmol/L,which were higher than (40.7 ±2.6)μmol/L,(43.2 ± 2.6) μmol/L and (53.4 ± 3.2) μmol/L of the 3-AB + SAP groups.There were significant differences in the levels of creatinine between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.279,3.073,P ＜ 0.05).The renal MPO activity of the SAP 3,6,12 hours groups were (0.69 ± 0.06) U/g,(1.07 ± 0.09)U/g and (1.42 ±0.13)U/g,which were higher than (0.57 ±0.05)U/g,(0.75 ±0.06)U/g and (0.89 ± 0.07) U/g of the 3-AB + SAP groups.There were significant differences in the renal MPO activity between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.066,4.012,P ＜ 0.05).The pancreatic pathological scores of the SAP 3,6 and 12 hours group were 6.50 ± 0.53,9.06 ± 0.66 and 11.75 ± 0.89,which were significantly higher than 4.25 ± 0.31,6.06 ± 0.51 and 7.57 ± 0.59 of the 3-AB + SAP group (t =3.631,3.598,5.147,P ＜ 0.05).The structure of the kidney was normal in the SO group.Congestive changes were observed in glomerulus of kidney,the renal tubular epithelial cell was necrosed,and luminal narrowing or occlusion,hemorrhage in the intercellular substance and inflammatory cell infiltration were observed in the SAP 12 hours group.The pathological changes of the 3-AB + SAP 12 hours group were significantly slighter (P ＜ 0.05).The relative expressions of poly ADP-ribose,ICAM-1 and P-selectin of the SO group were 1.00 ±0.21,1.00 ±0.18,1.00 ± 0.16,which were significantly lower than 3.83 ± 0.63,5.42 ± 0.83,3.71 ± 0.48 of the SAP 12 hours group (t =6.955,23.107,10.352,P ＜ 0.05).The relative expressions of poly-ADP-ribose,ICAM-1 and P-selectin of the 3-AB + SAP 12 hours group were 1.94 ± 0.36,2.35 ± 0.35,2.11 ± 0.29,which were significantly lower than SAP 12 hours group (t =3.977,12.115,5.012,P ＜ 0.05).Conclusions Poly ADP-ribose polymerase inhibitor 3-AB protects kidney from injury in the experimental SAP rats.Poly ADP-ribose polymerase inhibitor 3-AB functions by suppressing the ICAM-1 and P-selectin expression and reducing neutrophil infiltration.

SCLC can spare more volume of the normal lungs and e-sophagus, and has the ability of dose escalation.

Objective To investigate whether the change of beam set-up methods will influence the dosimetric quality of intensity modulated radiation therapy (IMRT) for non-small cell lung cancer (NSCLC).Methods Twenty-one stage Ⅰ-Ⅲ NSCLC patients were selected for this study.The technique of step and shoot was used and three different beam set-up methods were chosen for IMRT planning,including IMRT-7 with nine equal-spaced beams angled 0°,51°,102°,153°,204°,255°and 306°; IMRT-5 with five equal-spaced beams angled 0°,72°,144°,216°and 288°; and IMRT-5m which was created from IMRT-7 but excluded 2 fields (51°and 102° were omitted if there was lesion in the right lung,while 255°and 306° were excluded if there was lesion in the left lung).The dose constrains ofnormal lungs for IMRT were set according to V5-V60 of normal lungs obtained from the same patient's actually treated 3D-CRT dose volume histogram.The prescription dose for IMRT started from 65 Gy,and then escalated or decreased step by step by 2 Gy once a time until the best plan was obtained.Results For normal lung dose,IMRT-5m had lower V5-V25 than the other two groups; but there was no significant difference in V30-V40.IMRT-5 was the worst for V45-V60; and mean lung dose was lowest in IMRT-5m.Dose parameters of esophagus and spinal cord,target conformity index,and total monitor units were all similar among difference plans.IMRT-5m had lowest heart V40 compared to the other two groups.For target heterogeneity index,IMRT-5 was higher than IMRT-7,but there were no significant differences among IMRT-5m,IMRT-5 and IMRT-7.Compared to 3D-CRT,the prescription dose could be increased by (5.1 ±4.6) Gy for IMRT-7,(3.1 ±5.3) Gy for IMRT-5,and (5.5 ±4.8)Gy for IMRT-5m.Conclusion Fewer beams and modified beam angles could result in similar,even better plan quality.

AIM:Toinvestigatewhetherautophagyisup-regulatedwhenresveratrol(Res)inducesapoptosis in chondrosarcoma , and to study the effects of autophagy inhibitor combined with Res on chondrosarcoma .METHODS:SW1353 cells were divided into 4 groups: control group, Res group, 3-methyladenine (3MA) group, and Res +3MA group.Electron microscopy was used to observe the autophagyosomes in control group and Res group .At the same time, the viability of the cells in the 4 groups was detected by CCK-8 assay.TUNEL staining and Western blotting (for determi-ning the levels of cleaved caspase-3, Bax and Bcl-2) were used to reflect levels of apoptosis in all groups .The expression of autophagy-related proteins Beclin 1, LC3-Ⅱ and p62 was detected by Western blotting .RESULTS: Exposure of the cells to Res resulted in a decrease in cell viability and an increase in the level of apoptosis ( P<0.05 ) .Compared with control group, the level of apoptosis was increased but the autophagy was decreased (P <0.05).Compared with Res group, the cell viability and the level of autophagy were decreased and the level of apoptosis was increased ( P<0.05 ) . CONCLUSION:Resveratrol induces apoptosis and autophagy , and inhibition of autophgay enhances resveratrol-induced apoptosis in chondrosarcoma .

Objective To investigate the effect of lung protective and ventilatory management strategies for brain death donors on eligibility and availability of lungs for transplantation.Method The clinical data of two brain dead patients who accepted lung protective ventilatory management strategies from Feb.2015 to Mar.2015 were retrospectively analyzed.Two cases of brain-dead patients,due to severe cerebral trauma,accepted the aggressive lung protective ventilatory management strategies and airway management for 9 days and 4 days respectively.PaO2/FiO2,chest imaging manifestations,surface of the lung harvested and pulmonary rehabilitation of recipients after operation were observed.Result Two lung recipients were liberated from ventilation and pulmonary function improved significantly after double lung transplantation.Conclusion The application of lung protective ventilatory strategies in potential organ donors with brain death can increase the number of eligible and harvested lungs.

Objective Toinvestigatetheeffectsofpyrrolidinedithiocarbamate(PDTC)onexpressionofNF-κB,MMP-2andleft ventricular collagen remodeling following acute myocardial infarction in rats .Methods The myocardial infarction model in rat was induced by ligation of left anterior descending coronary artery .12 adult Sprague-Dawley rats survived 24 for h after acute myocardial infarction were randomly divided into the myocardial infarction (MI) group and the PDTC-treated(PD) group .Six rats were desig-nated as sham-operated group(SH group) .The PD group was intraperitoneally injected with PDTC (80 mg · kg -1 · d-1 ) for 28 d , the MI group and SH group were given normal saline as control .On 28 d ,the cardiac function of left ventricle was measured by ech-ocardiography .The infarct size was evaluated .The total collgen ,typeⅠcollgen ,typeⅢcollgen ,and Ⅰ /Ⅲ collgen ratio were quanti-fied histomorphometry .The mRNA and protein levels of NF-kappaBp65 and MMP-2 were determined by reverse transcription-poly-merase chain reaction(RT-PCR) and by Western blot ,respectively .Results Compared with the SH group ,the values of the total collgen ,typeⅠcollgen ,typeⅢcollgen ,and Ⅰ /Ⅲ collgen ratio in the MI group and the PD group were significantly increased ,the differen had statistical significance (P<0 .01) .The values of the total collgen ,typeⅠcollgen ,typeⅢcollgen ,and Ⅰ /Ⅲ collgen ratio in the PD group were notably decreased than those in the MI group(P<0 .01) .Moreover ,the mRNA and protein levels of NF-kap-paBp65 and MMP-2 in the PD group were lower than those in the MI group ,the difference had statistical significance(P<0 .01) . Conclusion Left ventricular collagen remodeling following acute myocardial infarction could be improved by PDTC to some extent , which mechanism could be related with inhibiting NF-kappaB activation and down -regulating the expression of MMP-2 in rats .

Objective To observe the changes of tissue morphology and ultrastructure of kidney in the rat model of acute necrotizing pancreatitis (ANP),and to investigate the protein expression of glycogen synthase kinase-3β(GSK-3β) and phosphorylated GSK-3βin renal tissue.Methods Sixty SPF male SD rats were randomly divided into 5 groups (n =12 for each group) according to random number method,including control group,ANP 3 h,6 h,12 h,24 h groups.ANP model was established by retrograde infusion of 5％ sodium taurocholate solution into the biliopancreatic duct.Rats were sacrificed at corresponding time points to collect pancreatic and left renal tissue.Serum amylase (AMY),lipase (LIPA),creatinine (Cr) and urea nitrogen (BUN) levels were detected.Pancreatic and renal tissues were routinely pathologically examined.Rephrocytes' ultrastructure changes were observed by projection electron microscope.GSK-3β protein expression and phosphorylated GSK-3β(p-GSK-3β) in kidney tissue were quantified by Western-blot.Results Serum AMY,LIPA,Cr,Bun and pathological scores for pancreatic and renal tissues in ANP groups were obviously higher than those in control group,which increased gradually with the progress of pancreatitis.In ANP rats,it was observed that the microvilli on the surface of the epithelial cells of renal tubules were swelling and irregularly arranged,the nucleus was condensed and broken,the nuclear chromatin was condensed and separated from the nuclear membrane,the mitochondria was condensed,swelling and vacuolated.The expression levels of GSK-3β protein in the renal tissue of the control group and ANP 3 h,6 h,12 h,24 h groups were 0.702± 0.044,0.876± 0.017,0.872± 0.034,0.855± 0.035 and 0.852± 0.032,respectively.The expression levels of p-GSK-3β were 0.626 ± 0.029,0.790 ± 0.029,0.616 ± 0.021,0.448 ±0.028 and 0.439 ± 0.017.GSK-3β protein expression was higher in ANP group than in control group,and the difference was statistically significant (all P ＜ 0.05).But there was no statistically significant difference at different time points in ANP group.p-GSK-3β protein expression increased at 3 h after modeling,and then gradually decreased.p-GSK-3β protein expression was higher in ANP 3 h group than control group and other ANP groups,which in ANP 12 h,24 h group was obviously lower than control group and ANP 3 h,6 h group,and the difference was statistically significant (P ＜ 0.05).Conclusions GSK-3β expression in the kidney of ANP rats began to increase at 3 h after modeling and maintain a high level.p-GSK-3β was transiently increased at 3 h after modeling and then gradually decreased to a level obviously lower than control group.It indicated that these changes may play a crucial role in ANP associated kidney injury.

Objective To explore the protective mechanism of glycogen synthase kinase-3β(GSK-3β) inhibitor TDZD-8 on acute necrotizing pancreatitis (ANP) associated kidney injury in rats. Methods SPF male Wistar rats were randomly divided into four groups (n = 20): sham operation group (Sham group), ANP model group, TDZD-8 intervention group and TDZD-8 control group. The rat ANP model was prepared by retrograde injection of 5% sodium taurocholate into the bile duct; the same volume of normal saline was injected into the pancreatic duct of the Sham group. The TDZD-8 intervention group and the TDZD-8 control group were injected with GSK-3β inhibitor TDZD-8 (1 mL/kg) via the femoral vein 30 minutes before the model or sham operation; the ANP model group and the Sham group were injected equal volume of 10% dimethyl sulfoxide (DMSO). Rats in each group were sacrificed at 12 hours after operation to measure the serum amylase (AMY), blood lipase (LIPA), serum creatinine (SCr) and blood urea nitrogen (BUN) levels and to observe the pathological changes of pancreatic tissues and kidney tissues. Ultrastructural change of renal cells was analyzed by transmission electron microscopy. Serum interleukin-1β (IL-1β) and interleukin-6 (IL-6) levels were evaluated by enzyme linked immunosorbent assay (ELISA). The activation of nuclear factor-κB p65 (NF-κB p65) was evaluated by immunohistochemistry assay. The protein expressions of GSK-3β, phospho-GSK-3β (Ser 9), tumor necrosis factor -α (TNF-α), inducible nitric oxide synthase (iNOS), intercellular adhesion molecule-1 (ICAM-1) and interleukin-10 (IL-10) in the kidney were determined by Western Blot. Results Compared with the Sham group, the serum and inflammatory factors levels of the ANP model group were significantly increased, the pathological damage of the pancreas and kidney tissues were severe, the histopathological score was significantly increased, the expression of NF-κB p65 was enhanced in the nucleus of the kidney tissue, and the expressions of GSK-3β, TNF-α, ICAM-1 and iNOS were significantly enhanced, and the expressions of p-GSK-3β(Ser 9) and IL-10 were significantly attenuated. Compared with the ANP model group, TDZD-8 pretreatment significantly reduced serum and inflammatory factor levels in the ANP model group [AMY (kU/L): 5.60±0.30 vs. 10.07±0.34, LIPA (U/L): 1 111.0±110.8 vs. 2 375.0±51.1, SCr (μmol/L): 47.38±1.48 vs. 72.50±2.43, BUN (mmol/L): 17.6±1.0 vs. 26.0±1.0, IL-1β (ng/L):195.90±5.50 vs. 332.40±38.29, IL-6 (ng/L): 246.10±26.74 vs. 385.30±32.19, all P < 0.01]; pathological damage of pancreas and kidney tissue (histopathological score: 7.1±0.4 vs. 12.1±0.3, 301.2±7.5 vs. 433.5±13.8, both P < 0.01) and ultrastructural damage of renal cells were alleviated; the expression of NF-κB p65 in the nucleus was significantly decreased; the expression of p-GSK-3β(Ser 9) was significantly increased, and blocking GSK-3β activity could inhibit the expressions of TNF-α, ICAM-1, iNOS and increase the expression of IL-10, while the expression of GSK-3β in renal tissues was not statistically significant. There were no significant differences between the TDZD-8 control group and the Sham group. Conclusions Blockade of GSK-3βactivity by TDZD-8 exerts the protective effect against kidney injury by inhibiting the inflammation signaling pathway in ANP. It can alleviate histopathological and ultrastructural changes in kidney injury, which protection mechanism is mediated by NF-κB and its related inflammatory mediators.