OBJECTIVE To investigate the effect of ventilation on the changes in TNF-? concentration of BALF on acute lung injury model.METHODS Forty healthy male Sprague-Dawley rats were randomly divided into five groups LV,HV,LV2P,LV5P,LV8P and the rat model of acute lung injury was built.Mechanical ventilation was employed in the test groups.Rats were sacrificed after 2 hours ventilation when the serum acquired.The supernatant of BALF was detected for TNF-? concentration.RESULTS The changes in TNF-? content of serum:the TNF-? contentration of serum was higher in HV group than that in LV group(P

Objective To investigate the relationship between self-management efficacy and medication adherence of the postoperative replacement therapy of thyroid cancer.Methods Of all the 230 thyroid cancer cases treated in the Thyroid Surgery Department of Ningbo No.2 Hospital from Jun.2014 to Jun.2016,125 cases were randomly chosen for the investigation by using the Chinese cancer self-management efficacy scale and the Morisky medication adherence questionnaire.Results The total postoperative alternative medication adherence rate was 47.28±11.49,among which the medium and low adherences accounted for 90.18％.The self-management efficacy score was 74.68±22.80 with scoring index of 62.58％ at the relatively medium level.Statistical significance was observed between the self-management efficacy and the postoperative alternative medication adherence (P＜0.01).Conclusion The self-management efficacy exerts significant effects on the postoperative alternative medication adherence of the thyroid cancer patients in promoting the postoperative recovery and improving the life quality.

Objective To investigate the relationship between the activity of Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in cerulein-induced pancreatic acinar cell line AR42J. Methods The in vivo model of AP was induced by cerulean treated pancreatic acinar cell line AR42J, then RPM and AG490 were given for intervention. Western blot was used to determine theexpressions of JAK1 and phosphorylation JAK1 ( P JAK1 ) , STAT1, PSTAT1 and TNFα, IL-1β, IL-6. The expressions of IL-6, IL-1 β, and TNFα mRNA were measured by RT-PCR. Survival rate of cells was evaluated by trypan blue stain. Results The relative expressions of JAK1, P JAK1, STAT1, P STAT1 and TNF-o, IL-1β, IL 6 without cerulean treatment were 0.09 ±0.04,0.14 ±0.08,0.21 ±0.09,0.12 ±0.12,0.10 ±0.02,0.08 ± 0.03,0.02 ± 0.02. After cerulean treatment, the expressions of abovementioned protein increased in a time-dependant manner, the expressions at 24h were 0.53 ± 0.09,0.53 ± 0.13,0.56 ± 0.09,0.55 ± 0.10,0.25 ± 0.04,0.25 ±0.09,0.27 ±0.07, which were significantly higher than those in the control group (P ＜0.05). 2 4 h after RPM and AG 4 9 0 inhibition, the expressions of TNF-α, IL-1 β, IL-6 proteins significantly decreased to 0.17 ± 0.03and 0.17 ± 0.01,0.15 ± 0.05 and 0.14 ± 0.07,0.19 ± 0.04 and 0.19 ± 0.05; their expressions of mRNA significantly decreased ( P ＜ 0.05 ). The cell survival rates in RPM and AG490 treatment group were (72.4 ± 11.2) %, (69.7 ± 9.8 ) %, and in cerulein-stimulated cells (42.2 ± 12.3 ) % ( P ＜ 0.05 ).Conclusions The JAK1/STAT1 signaling pathway was involved in pancreatic inflammatory response with cerulein stimulation. Early treatment with inhibitors to the JAK1/STAT1 signaling pathway might control the inflammatory response in acute pancreatitis.

Objective To study the protective effects of activated protein C (AFC) on the apoptosis of endothelial cells induced by lipopolysaccharide (LPS) in order to clarify the mechanisms associated with the expression of some genes related to apoptosis. Method The human umbilical vein endothelial cells were incubated with LPS (1.0 μg/mL) for one hour to make the models of cell apoptosis, and then the different concentrations of AFC (10 ng/mL and 50 ng/mL) were added to the models of cell apoptosis as treatment group. Therefore, there were two groups, model group and APC treated group. The factors related with apoptosis such as P53, Bax, Bcl-2, and caspase-3 mRNA or protein level were measured by using RT-PCR and Western blotting. Results Compared with LPS stimulated cells, the expressions of P53, Bax and caspase-3 mRNA and levels of protein were decreased and the expression of Bcl-2 mRNA and protein level were increased in APC treated cells particularly in APC 50 ng/mL treated cells (P <0.05). Conclusions The APC inhibits the apoptosis of HUVECs induced by LPS via regulating the mitochondrial-dependent apoptosis pathway, and it may become a novel therapeutic agent for infection disease.

AIM: To study the effect of activated protein C(APC) at different concentrations on apoptosis of human umbilical vein endothelial cells(HUVECs) induced by lipopolysaccharide(LPS).METHODS: The HUVECs were induced by LPS(1.0 mg/L) as apoptotic model that was administered by different concentration of APC(10 ?g/L or 50 ?g/L).Meanwhile,the control group and induced apoptosis group induced by LPS(1.0 mg/L) stimulation were also set up.The changes of cellular ultrastructures were observed under electron microscope.The DNA ladder and TUNEL fluorescent staining were measured in cells.Annexin-Ⅴ/PI double staining was used to measure the cell apoptosis rate by flow cytometry.Cell survival rate was measured by MTT assay.The proliferating cell nuclear antigen(PCNA) expression levels in cells were also measured by Western blotting to reflect the proliferation of the cells.RESULTS: There were significant apoptotic changes in the cells induced by LPS,but the apoptotic changes were reduced and apoptosis rates were decreased in the cells treated with APC.Meanwhile,cell survival rate and the protein levels of PCNA were increased after APC treatment,particularly at the concentration of 50 ?g/L,which showed difference when compared with those induced apoptosis group by LPS(P

Objective To explore the role of pancreatic cancer-derived microvesicles (MV) and their enclosed microRNAs (miRNA) in the pathogenesis of pancreatic cancer induced diabetes mellitus (DM).Methods The supernatants of three pancreatic cancer cell lines SW1990, BxPC3 and PANC1 were collected, and MV were isolated with gradient centrifugation.The entrance of MV into pancreatic islet cell line MIN6 was proved by Western blot assay and fluorescence-label method.The miRNA-19a levels were measured in MV and MV-free supernatants of three pancreatic cancer cells lines.The three experimental groups were MIN6 cells separately treated by MV derive from SW1990, BxPC3 and PANC1, and untreated MIN6 cells were assigned to the control group.The miRNA-19a levels as well as changes of glucose stimulated insulin secretion (GSIS) were measured.Afterward, pre-miRNA-19a and anti-miRNA-19a were transfected into MIN6 cells by liposome, and the effects of them on GSIS were observed.Results CD63 and AGO2 as the protein markers of MV and the entrance of MV from pancreatic cancer into pancreatic islet cell line MIN6 were detected by Western blotting.The miRNA-19a levels in MV and MV-free supernatants of SW1990, BxPC3 and PANC1 were (132.7±16.0), (32.8±4.3), (78.4±8.9),(22.6±3.3), (63.3±12.0) and (23.3±3.3) pmol/L, respectively, and the differences were statistically significant (t=10.44, 10.12 and 5.56,all P＜0.01).Compared to the MIN6 control group, the miRNA-19a levels of MIN6 treated by MV from SW1990, BxPC3 and PANC1 significantly increased, and the 2-△△Ctvalue was 2.02 ± 0.50, 1.80 ± 0.41 and 2.11 ± 0.59, respectively, and the differences were statistically significant (t=2.97, 2.77, 2.84;all P＜0.05).Stimulated with high glucose, the GSIS of pancreatic islet cells treated by SW1990, BxPC3 and PANC1 in three groups decreased, which were (103.73±16.49), (141.17±11.26), and (138.24±13.97) ng · mg protein-1 · h-1 MV, respectively, and that of control group was (256.24 ± 33.05) ng · mg protein-1 · h-1.The differences were statistically significant (t=4.13, 3.30 and 3.29, all P＜0.05).Compared with control group, GSIS of pre-miRNA-19a treated MIN6 remarkably decreased, which was (126.17± 62.87) ng · mg protein-1 ·h-1 and (316.72±91.87) ng · mg protein-1 · h-1 , and the difference was statistically significant (t=2.97, P＜0.05).GSIS of MIN6 cells transfected with anti-miRNA-19a was higher than that of control group, which was (697.47±77.62) ng · mg protein 1 · h-1 and (355.33 ±84.77) ng · mg protein-1 ·h-1 , and the difference was statistically significant (t=-2.97,P＜0.05).Conclusion The entrance of MV derived from pancreatic cancer into pancreatic islet cell line MIN6 may cause the dysfunction of insulin secretion an important signaling molecules, miRNA-19a.

Objective To investigate the effects of different tidal volumes and positive end expiratory pressures on cell apoptosis in lung tissue of rats with acute lung injury.Methods Forty healthy male Sprague-Dawley rats were randomly(random number)divided into five groups,namely low tidal volume group(LV,VT 8 mL/kg),high tidal volume group(HV,VT 30 mL/kg),low tidal volume group with PEEP 2cmH2O(LV2P,VT 8 mL/kg,PEEP 2 cmH2O),low tidal volume group and PEEP 5cmH2O (LV5P,VT 8 mL/kg,PEEP 5 cmH2O)and low tidal volume group and PEEP 8 cmH2O(LV8P,VT 8 mL/kg,PEEP 8 cmH2O).After intravenous administration of oleic acid(OA,0.1 mL/kg),the rat model of acute lung injury was made.Mechanical ventilation was employed in rats of the experiment groups.Rats were sacrificed and their whole lungs were taken after mechanical ventilation for 2 hours.The transferase d-UTP end labeling assay(TUNEL)was used to define the extent and distribution of apoptotic cells in bronchus and lung tissues.The level of caspase-3 protein was determined by immunohistochemistry.Results The apoptotic cells on both alveolar septum and bronchial epithelium obviously increased with high level of caaspase-3 protein in HV group.The number of apoptotic cells obviously decreased with decrease in caspase-3 protein after PEEP ventilation.These changes were more significant in LV5P than those in other groups(P ＜ 0.01).Conclusions The mechanical ventilation with low tidal volume and PEEP produces protective effects on lung from injury.The cell apoptosis plays an important role in the course of VILI.

Objective To explore the relationship between the severity of experimental acute pancreatitis (AP) and the expression of Myc interacting zinc finger protein 1 (MIZ1) in rat, to evaluate the value of MIZ1 for severity assessment .Methods Acute necrotizing pancreatitis ( ANP) model was induced by retrograde injection of sodium taurocholate into the pancreatic duct , and normal saline was used in control group.The rats were sacrificed at 6, 24, 48 h, and the blood and pancreas tissue was collected , and serum amylase level , C reactive protein ( CRP ) , TNF-α, IL-6 and MIZ1 were determined by ELISA .Pancreatic tissue was routinely examined by pathologist , and the MIZ1 protein expression in pancreatic tissue was measured by immunohistochemistry and Western blot .Results The serum amylase levels of control group and ANP group at 6, 24, 48 h were (449 ±40), (578 ±25), (1 021 ±205), (971 ±143)U/L, and the levels of CRP were (123 ±23), (169 ±25), (226 ±34), (229 ±24)mg/L;and the levels of IL-6 were (20.16 ± 4.11), (38.60 ±12.05), (52.33 ±6.77), (44.83 ±4.30)ng/L;and the levels of TNF-αwere (55.33 ±3.32), (82.8 ±5.26), (120.66 ±16.00), (108.33 ±12.17)ng/L;and the levels of MIZ1 were (5.51 ± 0.34), (3.44 ±0.56), (2.11 ±0.11), (2.41 ±0.43) ng/L.The pathologic scores of pancreas were (1.83 ±0.75), (6.00 ±1.67), (8.16 ±2.70), (9.33 ±1.50), and the expressions of MIZ1 in pancreatic tissue were 0.81 ±0.05, 0.53 ±0.07, 0.31 ±0.06, 0.21 ±0.08.Except for amylase level of ANP 6h group, other parameters of ANP group were significantly different with those of control group , and the parameters of ANP 24, 48 h group were significantly different with that of ANP 6 h group ( P<0.01), but there was no significantly different between ANP 24 and 48 h group.MIZ1 expression was negatively correlated with serum amylase level , CRP, TNF-α, IL-6 and pathologic scores of pancreas , and the difference was statistically signific ant (P<0.01).Conclusions The decreasing expression of MIZ1 is closely correlated with the severity of AP , and may be a potential marker for prognosis evaluation .

The prevalence of inflammatory bowel disease (IBD) in China is increasing year by year, however, the efficacy and safety of commonly used therapeutic methods are limited.Granulocyte and monocyte adsorptive apheresis (GMA) is one of the effective methods for treatment of IBD used abroad, however, there is still lacking of such research in China.Aims: To investigate the efficacy and safety of GMA in IBD patients.Methods: A retrospective study was conducted in 21 cases of IBD patients [13 cases with ulcerative colitis (UC) and 8 with Crohn's disease (CD)] who accepted GMA treatment from May 2013 to July 2014 at the Shanghai Rui Jin Hospital.All the cases were poor responders to 5-aminosalycylic acid (5-ASA) or steroid-refractory.The clinical data were collected, and the clinical activity index (CAI), endoscopic activity index (EAI), laboratory parameters including serum albumin (Alb), hemoglobin (Hb), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), leukocyte count and percentage of neutrophils, as well as the adverse effects before and two weeks after the end of GMA treatment were analyzed.Results: After GMA treatment, both CAI and EAI were decreased significantly in UC and CD groups as compared with those before treatment (P all <0.05).Among laboratory parameters, Alb was increased in UC group and CRP was decreased in both UC and CD groups after treatment (P all <0.05).No significant differences were found in other laboratory parameters in both UC and CD groups before and after treatment (P all >0.05).The treatment was well tolerated with no severe adverse effects.Conclusions: GMA is safe and effective for ameliorating clinical symptoms, attenuating intestinal mucosal injury and controlling active inflammation in IBD patient that has not responded to 5-ASA or steroid treatment.Prospective clinical studies with large samples are needed to confirm these findings.

Objective To investigate the effect of protein inhibitor of activated signal transducer and activator of transcription 1 ( PIAS1 ) gene silencing on the inflammatory response of rat pancreatic acinar cell lines AR42J with cerulein stimulation, to study its role in the pathogenesis of acute pancreatitis.Methods The siRNA targeting PIASI was designed, synthesized, transfected into AR42J cells by lipofectmine 2000.24 h later, cerulean was added and cultured for another 24 h.Subsequent AR42J cells with cerulein stimulation were divided into 4 groups: cerulein, liposome, negative-siRNA and PIAS1-siRNA groups.In addition, a group with PBS was as control group.The activity of p38 mitogen- activated protein kinase (p38MAPK) was detected by western blotting.TNF-α, IL-1β, IL-6, matrix metalloproteinase (MMP) 9 expression were analyzed by RT-PCR and western blotting, respectively.Results The expression of p38MAPK in PIAS1-siRNA, negative-siRNA, liposome, cerulein,and control group was 1.93 ±0.11, 1.22 ±0.10, 1.30 ±0.17,1.32 ± 0.21, 0.12 ± 0.02;while the expression of phosphorylated p38MAPK was 2.10 ± 0.25, 1.36 ± 0.20,1.26 ±0.15, 1.23 ±0.25, 0.58 ±0.48, the expression in PIAS1-siRNA group was significantly increased when compared with other groups (P＜0.05).The levels of TNF-α, IL-1β, IL-6, MMP-9 mRNA were 1.66 ±0.15,1.66 ± 0.15,1.90 ±0.01, 1.56 ±0.20 in PIAS1-siRNA group, while the expression of protein was 2.06 ±0.37,2.20 ±0.34, 1.80 ±0.10, 1.17 ±0.05, which was markedly higher than those in other group (P ＜0.05).Conclusions PIAS1 gene silencing could enhance p38MAPK activity, and improve inflammatory mediator expression in pancreatic acinar cells with cerulein stimulation.