Objective To assess the characteristics of biochcmical parameters of bone metabolism in patients with inflammatory bowel disease (IBD) and the relationship between osteoporosis and the reduction of bone mass so as to identify the risk factors for osteoporosis in IBD. Methods Sixty-nine patients with IBD[49 with Crohn disease (CD) and 20 with ulcerative colitis (UC)] and 20 sex- and age-matched healthy subjects (control group) from the medical examination centre in 2007 were enrolled in the study. The disease activity was estimated according to CD active index and Truelove-Witts score. The biochemical markers of bone metabolism including ostecalcin (OC), 25-bydroxycholecalciferol[-25 (OH)Ds-] and cross-linked N-telopeptides of type I collagen(NTX) were tested using enzyme immunoassay. The body mass index (BMI) and bone mineral density (BMD) were also measured. Results The concentration of 25(OH)D3 was significantly lower in both CD[(44.45±39.38) nmol/L] and UC patients[(34.67±23.79) nmol/L] compared with controls[(98.42±25.84) nmol/L] (P <0.05), while NTX level was significantly higher in both CD [(58.41±15.15) nmol BCE/mmol Cr] and UC[(57. 67±10. 75) nmol BCE/mmol Cr] patients compared with controls[(30. 38±13.35) nmol BCE/mmol Cr] (P<0.05). The levels of OC and 25 (OH)D3 in patients treated with steroids[(17. 3±13. 43) ng/ml and (26. 99 ± 9. 12) nmol/L, respectively] were lower than those in patients untreated with steroids[(27.33 ± 16.86) ng/ml and (45.33±39.03) nmol/L,respectively] . BMD examination revealed that the incidence of osteoporosis in CD or UC groups were 14.3%(7/49) or 3/20,respectively, with no difference between two groups (P>0. 05). The T score of lumbar spine in patients treated with steroids (-1.19±0. 93) was significantly lower than that in patients untreated with steroids (-0.80±1.29) (P<0.05), but there was no difference in T score of femoral between two groups (P>0.05). Stepwise regression analysis revealed that advanced age and reduced BMI were risk factors for osteoporosis in CD patients.Conclusions The lower BMI in patients with IBS contributes to prevalence of osteoporosis, and advanced age and reduced BMI are risk factors for osteoporosis in CD patients. NTX and 25 (OH)D3are important bone metabolic markers to predict osteoporosis in patients with IBD.

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 investigate the pathologic changes in the pancreas of rats after intraperitoneal injection of DETC,a kind of superoxide dismutase (SOD) inhibitor,and to compared that with another model of chronic pancreatitis by pancreatic duct injection of TNBS.Methods The rats were randomly divided into DETC group,DETC control group,TNBS group,TNBS control group,normal control group.Rats in DETC group received an intra-peritoneal injection of DETC twice a week,and rats in DETC control group received an intra-peritoneal injection of same amount of normal saline.Rats in TNBS group was injected with 2％ TNBS ethanol phosphate buffer into the pancreatic duct,while rats in TNBS control group was treated with injection of same amount of ethanol phosphate buffer,and rats in normal control group received no treatment.The rats were sacrificed after 2 w,4 w,6 w and 8 w.The serum levels of amylase were determined,and pathological and ultrastructure changes of the pancreas were measured.The levels of SOD,GSH-PX activity and MDA content were detected.The expressions of α-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN in tissue were detected by immunohistochemical assay.The TGF-β1 mRNA expression was detected by RTPCR.Results No rat died in DETC group.The mortality rate of TNBS group was 15％.The serum levels of amylase were not statistically different between the 2 groups.The fibrosis scores of rat in DETC group at 4 w was 3.4 ± 1.l,which was significantly higher than that in TNBS group (3.0 ± 1.3,t =3.462,P ＜ 0.05).At 6 w,the damage scores of rat in DETC group was 9.1 ± 1.8,which was significantly higher than that in TNBS group (8.4 ± 1.8,t =2.943,P ＜ 0.05).Scores of vacuolar degeneration and fatty infiltration of rat in DETC group were higher than those in TNBS group,but the difference between the two groups was not statistically significant.Two weeks later,ultrastructure changes of pancreas could be observed,and large amounts of regenerative or mature collagen could be seen at 4 w.The SOD activity of DETC group was significantly decreased when compared with those in TNBS group (t =5.468,P ＜ 0.01).The GSH-PX activity of DETC group at 2 w,6w was significantly decreased when compared with those in TNBS group (t =6.497,10.125,P＜0.01).While the activity of MDA at 6 w,8 w was significantly increased when compared with those in TNBS group (t =3.350,5.407,P ＜0.05).The differences at other time points were not statistically significant.The expressions of (a)-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN,and TGF-β1 mRNA were not statistically significant between the 2 groups.Conclusions Sustained suppression of SOD activity can successfully induce chronic pancreatitis.Fatty infiltration and fibrosis in pancreas in DETC group occurs earlier with more severe presentation than that in TNBS group.Intraperitoneal injection of DETC is easy with low mortality rate,which is an ideal method for chronic pancreatitis model induction.

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.

Objective To investigate the status of Helicobacter pylori(Hp)infection with subclinical hepatic encephalopathy (SHE) and the level of blood ammonia. Methods Sixty-five SHE patients and healthy volunteer were selected. In all subjects Hp was assessed by rapid urease test (RUT) and histological examination. It would be regarded as Hp infection if both the RUT and histological examination were positive. The level of blood ammonia was detected and number connection test (NCT), digit symbol test (DST)and electroencephalogram (EEG)was performed. The patients could be diagnosed with SHE whichever of the three tests was abnormal. The patients with Hp infection were randomized received one-week standard treatment. Bacterial eradication was assessed with <'14>C-urea breath test after the treatment of 6-8 weeks, meanwhile above mentioned examinations were reassessed. Results The ratio of Hp infection in SHE patients (46.2%, 30/65 )was higher than that in healthy controls ( 33.3 % ,20/60), but there was no significant difference(P> 0.05).The blood ammonia level before and after eradication had no significant difference(P> 0.05 ). No improvement was observed in above mentioned examinations after treatment. Conclusion There is no relationship between the level of blood ammonia and Hp infection in SHE.

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 .

Objective To investigate the effect of antioxidants including PDTC on pancreatic fibrosis of rats with chronic pancreatitis.Methods The rats were randomly divided into 5 groups including control group, CP group, PDTC treatment group, vitamin E treatment group and vitamin C treatment group.The CP model was in ducad by using intraperitoneal injection of DETC (750 mg/kg), twice a week.The control group received no treatment.After DETC injection, the treatment groups received an intraperitoneal injection of PDTC (100 mg/kg), vitamin E (15 mg/kg), vitamin C (15 mg/kg), respectively.Rats were sacrificed at 90 min, 24 h, 48 h, 72 h, 2 w, 3 w, 4 w, 6 w after first injection of DETC.Pancreatic tissue was taken for routine pathological examination.The activity of SOD, GSH-PX and MDA content were detected by spectrophotometric ratio method.α-SMA, desmin collagen Ⅰ, Ⅲ, TGF-β1, FN were detected by immunohistochemical assay.The expression of TGF-β1, FN mRNA was measured by RT-PCR.Results At 6w, the fibrosis and the parameters for damage of the pancreas in the three treatment groups were significantly better than that in CP group (P ＜0.01), the vacuolar degeneration index in vitamin E group and vitamin C group was also better than that in CP group (P ＜0.01).From the 2nd week, the activity of SOD, GSH PX in PDTC group, Vit C group and Vit E group was higher than that in CP group, while the MDA activity was lower than that in CP group, and the difference was statistically significant (P ＜ 0.01 or P ＜ 0.05).No significant difference was found among the three treatment groups.The mRNA levels of TGF-β1 and FN of the treatment groups were lower than those of CP group (P ＜0.05 or P ＜0.01), but higher than those of the control group (P ＜ 0.05).There was no significant difference among the three treatment groups (P ＞ 0.05).Conclusions PDTC and the other antioxidants can reduce oxygen free radicals by increasing the activity of SOD,suppressing the activation of PSCs, reducing the secretion of TGF-β1, Collagen Ⅰ , Ⅲ, FN and eventually inhibit the progress of pancreatic fibrosis.

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

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 investigate the role of connexin 43(Cx43)in the apoptosis of pancreatic cancer cell line BxPC and its possible mechanism.Methods pcDNA-Cx43,pcDNA-Cx43N,pcDNA3.0,siRNA-Cx43 and siRNA-NC were transfected into BxPC3 cells via liposome method.Cx43 protein and Cytochrome C(Cyt C)concentration was determined by Western blot,and the apoptosis was analyzed by Annexin V/PI binding assay.The mitechondria apoptosis pathway involved in Cx43 associated apoptosis was examined which contains the depolarization of mitechondrial membrane potential (MMP);fluorospectrophotometer was used to measure the activities of caspase-3 and caspase-9. Gap junction intercellular communication(GJIC) was determined by dye-transfer method.Results Cx43 protein expression increased after BxPC3 transfeetion,apoptosis rate increased from(6.35±0.43)%in empty vector transfection group to(14.29±1.24)%;after H202 treatment,apoptosis rate increased from(20.34±2.47)%to(31.27±2.56)%(P<0.05).Meanwhile,mitochondrial membrane potential was decreased,Cyt C was increasingly released from mitochondria,caspases activities were increased;after siRNA43 interference,apoptosis rate decreased from(7.42±0.47)% to(5.19±1.37)%,after H_2O_2 treatment,apoptosis rate decreased from (19.43±1.71)%to(11.67±1.97)%(P<0.05).Decreased mitochondrial membrane potential and Cyt C release were observed,caspases activities were decreased.GJIC of pcDNA-Cx 43 transfection group increased from 14.52±0.57 to 23.05±3.84.and it increased from 1.70 ±0.24 to 3.84 ±0.45 in the presence of β-GA(P<0.05).But the apoptosis rate was not significantly different.Conclusions Cx43 could promote BxPC3 apoptosis via mitochondrial apoptotic signal pathway,and the possible mechanism included signal pathway other than GJIC.