Objective To investigate the effects of pioglitazone pre-treating on pancreatic acinar cell (AR42J cells) apoptosis induced by caerulein.Methods AR42J cells were divided into blank control group (with normal culture),pioglitazone group (40 μmol/L),caerulein control group (1 × 10-8 mol/L),pioglitazone+ caerulein group (40 μmol/L pioglitazone + 1 × 10-8 mol/L caerulein) and pioglitazone + GW9662+caerulein group (40 μmol/L pioglitazone+ 5 μmol/L GW9662 + 1 × 10-8 mol/L caerulein).Pioglitazone and GW9662 were added 30 minutes earlier than caerulein.Cell proliferation rate of each group was determined by MTT assay at three,six,12 and 24 hour.The cell apoptosis rate was detected by flow cytometry with Annexin Ⅴ/PI staining and terminal dexynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) staining.The activity of Caspase 3,8 and 9 of each group was measured.Mitochondrial membrane potential (MMP) was detected by flow cytometry with JC-1 staining.Single factor analysis of variance and LSD test were performed for data analysis.Results At six,12 and 24 hour,the cell proliferation rate of pioglitazone group and pioglitazone + caerulein group was 0.19±0.02,0.22±0.02,0.36±0.02 and 0.20±0.04,0.23±0.02,0.38±0.02,respectively,which were significantly lower than those of blank control group (0.25 ±0.04,0.28 ± 0.03 and 0.46±0.02) and caerulein group (0.23±0.02,0.29±0.01 and 0.46±0.05,t lgroup=-3.16,-4.61 and-6.25,tcaerulein group =-1.58,-4.61 and-6.15,all P＜0.05).And the cell proliferation rates of pioglitazone+GW9662+caerulein group at six,12 and 24 hour (0.23±0.02,0.27±0.02 and 0.45±0.01) were significantly higher than those of pioglitazone+caerulein group (t=2.25、3.87、4.56,all P＜0.05).There was no significant difference in cell apoptosis rate detected by flow cytometry with Annexin Ⅴ/PI staining between pioglitazone group ((11.80 ± 0.47) ％,(9.62 ± 2.63) ％ and (14.92 ± 2.52) ％) and pioglitazone+caerulein group ((8.78±0.47)％,(11.89±2.80)％ and (14.25±2.67)％,all P＞0.05),but cell apoptosis of these two groups were higher than those of control group ((5.52± 0.64)％,(5.30±0.97)％ and (5.47±0.88)％) and caerulein group ((5.98±1.21)％,(7.47± 0.58) ％ and (8.11 ± 1.32) ％) respectively,and the differences were statistically significant (t l group =9.81,4.45 and 10.74,tcaerulein group =4.38,7.62 and 6.98,all P ＜0.05).There was no significant difference in apoptosis rate between pioglitazone+GW9662+caerulein group ((5.82±0.26) ％,(6.05± 0.83) ％ and (9.23±0.90)％) and caerulein group; while significantly higher when compared with those of pioglitazone+ caerulein group (t=-4.63,-10.07 and-5.70,all P＜0.05).At 12 hour,the apoptosis rate detected by TUNEL staining of pioglitazone group ((3.93 ± 0.40)％) was significantly higher than that of control group ((2.73 ±0.68) ％),the apoptosis rate of pioglitazone+ caerulein group ((8.43 ± 1.65)％) was significantly higher than that of caerulein group ((2.80 ± 0.56)％),the apoptosis rate of pioglitazone+GW9662+caerulein group ((3.87±0.35)％) was lower than that of pioglitazone+ caerulein group (t=7.93,8.92,-5.35,all P＜0.05).At 24 hour,the activity of Caspase 3,8 and 9 of pioglitazone+ caerulein group (1.28 ± 0.05,1.38 ± 0.04 and 1.53 ± 0.09) significantly increased compared with those of caerulein group (1.12±0.88,1.22±0.02 and 0.53±0.07,t=3.20,8.62 and 1.29,all P＜0.05).After treated with GW9662,part of activity of Caspase enzymes recovered.The number of cells with potential change of mitochondrial membrane in pioglitazone group and pioglitazone + caerulein group was more than that of caerulein group (28.50±0.91)％ and (28.20±2.56)％ vs (15.00±3.67)％) and part of membrane potential recovered after GW9662 added ((20.67 ± 2.20) ％).Conclusions Pioglitazone might promote AR42J cell apoptosis through the activation of caspases enzymes and changing membrane potential.And the antagonist GW9662 would partially inhibit the apoptosis induced by pioglitazone.

Objective To investigate the effect of pioglitazone on the activation of pancreatic apoptosis in the pathogenesis of rats with acute necrotizing pancreatitis.Methods Eighty Sprague-Dawley (SD) rats were randomly divided into four groups,including acute necrotizing pancreatitis (ANP),sham operation (SO),solvent control (Solvent),pioglitazone intervention (pioglitazone) group,with 20 rats in each group.ANP model was induced by retrograde injection of 4％ sodium taurocholate (1ml/kg body weight) into the biliary-pancreatic duct.The rats in pioglitazone group were injected pioglitazone (40 mg/kg body weight) into the ANP abdominalcavity 30 min before mldel induction.The rats were sacrificed at 1 h,3 h,6 h,and 12 h after ANP model induction.The pancreatic tissues were harvested.Routine HE staining was used to evaluate pancreatic pathological damage.The apoptosis was determined by TUNEL method.The expression of PPARγ was determined by using immunohistochemistry and Western-blot methods.The activity of caspase3 in pancreatic tissues was detected by using spectrophotometry.Results The pancreatic pathological damage was attenuated in rats in pioglitazone group compared with that in rats of ANP group,and the difference between the two groups was statistically significant (P ＜ 0.05).The PPARγ expression of pioglitazone group was 1.34 ± 0.09,which was significantly higher than that in ANP group (0.75 ± 0.05),and the difference between the two groups was statistically significant (P ＜ 0.05).The apoptotic index in pioglitazone group at 3 h was 8.35 ± 0.95,which was significantly higher than that in ANP group at 3 h (4.37 ± 1.22) ; the caspase3 activity was 9.24 ± 1.78,which was significantly higher than that in ANP group (5.04 ± 0.86),and the difference between the two groups was statistically significant (P ＜0.05).Conclusions Pioglitazone intervention attenuates pancreatic inflammation,increases PPARγ expression and caspase3 activity and induces apoptosis in pancreas of rats with acute necrotizing pancreatitis.

Objective To study the relation between the effect of Curcumin on the proliferation and activation of hepatic stellate cells(HSC)and the expression of the peroxisome proliferator-activated receptor?(PPAR?).Methods The rat HSC was isolated from SD rats through in situ perfusion of liver with Pronase E and density-gradient centrifugation with Nycodenz.The subcultured cells were treated with corresponding compound.The inhibition effect on HSC proliferation was determined by MTT colorimetry.The total RNA was extracted by TRizol reagent,and the gene expression level of PPAR? and smooth muscle actin (?-SMA)were determined by semi-quantitative RT-PCR.The total cellular proteins were extracted and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis,and the protein level of ?-SMA was determined by Western blotting.Results The MTT analysis results showed that Curcumin inhibited HSC proliferation between 10 and 50 ?mol/L in a dose-dependent manner.In the Day 1,4,7 of primary HSC and passaged HSC,the expression of PPAR? mRNA decreased with HSC activation extent;Curcumin could up-regulate the expression level of PPAR?.Curcumin suppressed the expression of ?-SMA not only at the gene level but also at the translation level.But all these effects of Curcumin on HSC could be blocked by the PPAR? specific antagonist GW9662.Conclusion The effect of Curcumin on HSC cell proliferation and activation is through the up-regulation of PPAR? signal.

Aim To study the effect of Curcumin on the apoptosis of hepatic stellate cells (HSC), and the correlation between the effect and peroxisome proliferator-activated receptor ?(PPAR?) signal.Methods The HSC was isolated from normal SD rats through in situ perfusion of liver with protease E and density-gradient centrifugation with Nycodenz.The subcultured cells were treated with corresponding compounds. Cell apoptosis was detected by Hoechst 33258 staining. PPAR? subcellular distribution was detected by immunofluorescent staining. Total RNA, total protein and nuclear protein were extracted respectively, target gene and protein levels were determined by semi-quantitative RT-PCR or Western blot.Results There was nearly no apoptosis in activated HSC. Curcumin treatment induced the apoptosis of HSC, enhancing PPAR? nuclear translocation/redistribution.At the transcription and translation level,curcumin upregulated nuclear PPAR? expression, inhibited anti-apoptotic Bcl-2 expression, and promoted pro-apoptotic Bax expression; but all these effects could be reversed by PPAR? antagonist GW9662.Conclusions Curcumin induces HSC apoptosis by enhancing PPAR? expression and nuclear translocation/redistribution.

Objective:To study the effect of continuous blood purification on inflammatory state and immune response in patients with multiple injury and sepsis.Methods:88 patients with multiple injury and sepsis who were treated in our hospital from January 2015 to May 2016 were selected as the research object,they were divided into the control group (n=44) and observation group (n=44) according to random number table method.The control group was given conventional treatment,and the observation group was given continuous blood purification treatment.The acute physiology and chronic health conditions Ⅱ (APACHE Ⅱ)score was observed in two groups Ⅱ at 3 d after treatment.The serum content of Interleukin-2(IL-2),Interleukin-4(IL-4),Interleukin-10(IL-10) and interferon-γ (IFN-γ) were detected by ELISA method.The levels of CD3 +,CD4 +,CD8 + and NK cell were analyzed by flow cytometry.The urea nitrogen (BUN),serum creatinine (Scr),prothrombin time (PT),thrombin time (TT),activated partial blood coagulation time (APTT) were tested by automatic biochemical analyzer.Results:The APACHE Ⅱ score,BUN and Scr levels in observation group were lower than the control group (P＜0.05).The content ofIL-2,IL-4,IL-10 and IFN-γ in observation group were lower than the control group (P＜0.05).The levels of CD3+,CD4+,CD8+ and NK in observation group were higher than the control group (P＜0.05).The levels ofPT,TT and APTT in observation group were lower than the control group (P＜0.05).Conclusion:Continuous blood purification in patients with multiple injury and sepsis had better clinical curative effect,can reduce inflammation,improve immune function and the function of blood coagulation.

Objective To observe the expression of caspase-3 on the trehalose as cryoprotectant for preserving aortic valve homograft in liquid nitrogen.Methods The aortic valve homograft was divided into 5groups,namely:0.1 mol/L DMSO(control group),0.1 mol/L trehalose(experimental group 1),0.1 mol/L trehalose+0.1 mol/L DMSO(experimental group 2),0.2 mol/L trehalose+0.1 mol/L DMSO(experimental group 3),0.3 mol/L trehalose+0.1 mol/L DMSO(experimental group4).At the time of 12 months,15 months and 18 months when preserved in liquid nitrogen,relative expression of caspase-3 of the aortic valve homograft was measured by RT-PCR and Western Blot.Fresh group was a negative control group.Results At the same time(P<0.05),the expression of caspase-3 of fresh aortic tissue was slightest.The experimental group 2 was in accord with the experiment group 3,which was of a sort compare with the fresh group.The experimental group 4,which was worse than the experimental group 2 and 3,ranked above the experimental group 1.The worst was the control group.Conclusions The joint use of trehalose and DMSO could well inhibit the expression of caspase-3.Moreover.0.1mol/L trehalose+0.1 mol/L DMSO and 0.2 mol/L trehalose +0.1 mol/L DMSO could maximize the inhibition of the expression of caspase-3.

Objective To evaluate tumor angiogenesis and clinical significance by contrast-enhanced CT in case of non-small cell lung cancer (NSCLC). Methods 30 patients with NSCLC underwent dynamic thoracic CT, and histopathological slides were carefully prepared for VEGF immunohistochemical staining. Maxium attenuations of dynamic CT were compared with VEGF expression levels and lymph-node metastases. Results The mean peak attentation was (36.28?6.41)HU, VEGF positive expressions were in 21 patients, and negative expressions were in 9 patients. VEGF expression levels in patients with NSCLC stage II and III were higher than those in stage I , and in lymph node metastases group, the expression levels of VEGF were also higher than those in non-metastatic nodes. CT enhancement of NSCLC was positively related to VEGF expression,neoplasm stage and lymph-node metastases. Conclusion CT enhancement of NSCLC can reflect tumor angiogenesis and correlate to lymph-node metastases closely, help lung cancer diagnoses,neoplasm stage, and serve as a supplement to the present staging system for lung cancer in biological behavior.

Objective To study the clinical significance of P - selection expression in children with viral encephalitis and the correlation between this expression and the cerebral infarction with critical viral encephalitis. Methods Flow cytometric was employed to detect the expression of P- selection on the surface of platelet membrane in 44 children with viral encephalitis(20 light patients and 24 critical patients) and 20 healthy control children. The area of the cerebral infarction was determined by computed tomographic scan in 20 patients with critical viral encephalitis. The correlation between the two variables was analyzed. Results The expressions of P - selection on the surface of platelet membrane on less than 5 days and on 2 weeks after the onset of viral encephalitis were significantly higher in critical patients than those in normal control children and light patients( P

Objective To explore the effect of circadian rhythm disorder on cognitive decline and neurologic impairment after traumatic brain injury (TBI) in rats.Methods Rats were subjected to a weight-drop model of TBI,then the rat models were divided into 2 groups ac-cording to the environmental length of day and night ,namely,the whole day group (12 h light/12 h light) and the control group (12 h light/12 h dark).After 14 days,the Morris water maze test and step-down test were carried out to evaluate the memory of the rats in each group . HE staining and immunohistochemistry ( BrdU, Ki-67) were carried out to evaluate the degree of the neurologic impairment of the rats in each group.And the mRNA expressions of Caspase-3,Caspase-9,Bcl-2 and Bax were evaluated with realtime PCR .Results The memory function of the rats in whole day group was significant lower than control group ;the damage degree of the cells in the hippocampus and the cortical lesion volume in whole day group were significant higher than control group ;the cell proliferation rate and newborn cell survival rate in the whole day group decreased significantly compared with the control group ;the mRNA expression of Bcl-2 in whole day group was signifi-ant lower than control group ,and the mRNA expression of Caspase-3,Caspase-9 and Bax in whole day group was signifiant higher than control group.Conclusion Circadian rhythm disorder can worsen TBI-induced cognitive decline and neurologic impairment .

Objective To observe effects of tetrandrine on β-glucan induced RAW 264 .7 cells proliferation .Methods RAW 264 .7 cells model was established .Four methyl thiazol tetrazolium(MTT) was used to detect the effects of different concentrations of tetrandrine on the proliferation of RAW264 .7 cells .The levels of interleukin(IL)-6 ,tumor necrosis factor-α(TNF-α) ,prostaglan-din E2 (PGE2 ) and IL-10 in the culture supernatant were measured by enzyme linked immunosorbent assay(ELISA) .Results MTT results showed that the growth curves of different concentrations of tetrandrine on RAW264 .7 cells had biphasic affections .ELISA results suggested that tetrandrine could inhibit the IL-6 ,TNF-α,PGE2 expressions and promote IL-10 expression .Conclusion Effects of tetrandrine onβ-glucan induced RAW264 .7 cells proliferation are relative to inhibition of IL-6 ,TNF-α,PGE2 expressions and promotion of IL-10 expression .