Objective To observe the influence of silent information regulator factor 1(SIRT1)on TNF-αinduced intestinal epi-thelial Caco-2 cell barrier function destroy and to investigate its molecular machenism.Methods Caco-2 cells were randomly divided into three groups:normal control group (control),TNF-αgroup (TNF-α,100 ng/mL for 24 h)and 100 ng/mL plus 40μm resvera-trol group (TNF-α+Res).Transepithelial electrical resistance (TER)was determined.SIRT1 and the protein expressions of ZO-1 , occludin were examined by using Western blot.Results The relative expression amounts of SIRT1 protein were 0.81 ± 0.02, 0.43±0.04 and 0.60±0.03 respectively.TER of three groups were (154.00±5.00),(97.00±4.00)and(128.00±6.00)Ohm/cm2 respectively.Compared with the control group,the expression of SIRT1 protein was reduced by 47% and TER was decreased by 37.00% in the TNF-αgroup.After resveratrol precondition,TER was increased by 32.00% compared with the TNF-αgroup. The relative expression amounts of ZO-1 and occludin protein in the control group,TNF-αgroup and TNF-α+Res group were (0.62±0.06,0.57±0.03),(0.23±0.05,0.33±0.04)and(0.41±0.03,0.50±0.02)respectively.After TNF-αtreatment,the ex-pressions of ZO-1 and occludin protein were significantly deceased(P<0.05),but the resveratrol precondition could attenuate this phenomenon,compared with TNF-αgroup,the protein expression was increased by 78.00% and 51.00% respectively (P<0.05). Conclusion Under the condition of TNF-αtreatment,the SIRT1 level is decreased,but increasing SIRT1 level could increase the in-testinal tight j unction protein ZO-1 and occludin protein expression,thus alleviate the damage of TNF-αon the epithelial barrier function constituted by Caco-2 cells.

Objective To study the effect of erythropoietin（EPO） on PI3-K/Akt signal pathway of myocardial ischemia-reperfusion injury（I/R） in immature rabbits. Methods Thirty immature rabbits were randomly divided into two groups （n = 15/greup ）, The rabbits were intraperi- toneally injected with 2 ml of normal saline in control group, or 5 000 U/kg of EPO in treated group 24 h before I/R. The Langendorff isolat- ed perfused heart model was established, and the myocardial expression of PI3K, AKT and Caspase 9 were examined by immunohistochem- istry in 2 h after repeffusion. To monitor the myocardial apoptosis,we performed TUNEL in 2 h after reperfusion. Results The expression of myocardial PI3K and AKT in EPO group was significantly higher in EPO-treated group than those in control group. In EPO-treated group, the expression of myocardial caspase 9 was significantly lower than that in control group. The average optical density of myocardial PI3K, AKT and caspase 9 were significantly different between both groups （P 〈 0.01 ）.The number of apoptotic cell was significantly fewer in EPO- treated group than that in control group. The myocardial apoptosis index （AI）was significantly different between two groups （P 〈 0.01 ）. Conclusion EPO pretreatment can influence the PI3-K/Akt signal pathway and decrease the myocardial apoptosis,and significantly reduce the myocardial ischemia-reperfusion（I/R） injury in immature rabbits.

Object To investigate the stability of baicalin in different conditions Methods The stability of baicalin in water with different pH values, methanol, ethanol, chloroform and in the mouse lung homogenate, mouse liver homogenate, mouse plasma and bovine serum was investigated by the classical isothermal method Results The experiments showed the baicalin was stable in acidic water and these organic solutions, but unstable in basic water and these biological media Conclusion The stability of baicalin in different media was different, which should be considered in practice

Objective: In order to provide an effective reference to the study on its preparation, the stability in vitro of andrographolide was investigated. Methods: Andrographolide was dissolved in phosphate buffer solutions of different pH values; classical isothermal method was employed; the content of andrographolide was determined by RP HPLC; then the stability of the drug in different biological media and organic solvents at 37?C was investigated. Results: The stability of andrographolide was different at different pH values and was best in the condition of pH=3～5. In biological media, it was most stable in bovine serum and the next was in mice homogenate. Conclusion: In different conditions, the stability of andrographolide was different. It provided a reference to the study on andrographolide and its preparation.

Objective To examine the effects of different doses of midazelam on ED50 of emulsified isoflurane and to determine the type of interaction between them for hypnosis by isobelographic analysis in rats. Methods One hundred and twenty-five adult male SD rats weighing 240-300 g were randomized into 5 groups (n=25 each): group Ⅰmidazolam (M); group Ⅱ emulsified isnflurane (Ⅰ); group Ⅲ ,Ⅳ ,Ⅴ, 1/4, 1/2 and 3/4 ED50 of midazelam for hypnosis + emulsified isoflurane (MI1 , MI2, MI3). Up-and-down sequential experiment was used to determine ED50of midazolam and emulsified isoflurane for loss of righting reflex in group Ⅰ -Ⅴ . The intial dose of midazolm was 17.3 mg/kg in group M. The initial doses of emulsified isoflurane (120 mg/ml) were 0.55 (in group Ⅰ, 0.22 (MI1), 0.19 (MI2) and 0.12 ml/kg (MI3) respectively. In group MI1-3 midazolam was injected over 15 seconds and after an interval of 2.5 min emulsified isoflurane was injected over 10 s. ED50 was calculated using Dixon-Mood method. Isebolographic and algebraic analyses were used to determine the type of interaction between midazolam and emulsified isoflurane. Results The five groups were comparable with respect to M/F sex ratio and body weight. The Edso of midazolam was 26.0 mg/kg in group M. Midazolam 6.5, 13.0 and 19.5 mg/kg were given in group MI1 , MI2 and MI3 respectively. The ED50 of emulsified isoflurane was 0.67 (ingroup Ⅰ), 0.30 (MI1), 0.22 (MI2) and 0.18 ml/kg (MI3) respectively. The isobolographic analysis indicated that with increasing doses of midazolam, the Edw of emulsified isoflurane decreased progressively in a non-linear fashion. The isobolographic and algebraic analyses demonstrated that the interaction between midazolam and emulsified isoflurane was synergistic for hypnosis. Conclusion The hypnosis is synergistic when midazolam 6.5,13 mg/kg are combined with emulsified isoflurane and additive when midazolam 19.5 mg/kg is combined with emulsified isoflurane.

Objective To observe the relationship between early lactate clearance and APACHEⅡ in critically ill patients. Methods In 121 critically ill patients,APACHE Ⅱ and lactate clearance rate were recorded. According to APACHE Ⅱ score,all patients were divided into≤10 score group, (11 ～20) score group,(21 ～30) score group and ＞ 30 score group, then compared the level of the early lactate clearance rate. The early lactate clearance rate were also compared between survival group and death group. Then the relationship between early lactate clearance and APACHE Ⅱ were analyzed. Results In( 11 ～20) score group,the early lactate clearance rate was lower than those in ≤ 10 score group, but the difference was not significant ( P ＞ 0. 05 ). The early lactate clearance rate in (21 ～ 30) score group ( 18. 35 ± 10. 01 ) % was lower than those in ( 11 ～ 20) score group (27.35 ± 10. 22) % ( t = 3.481, P ＜ 0. 01 ),in ＞ 30 score group( 11.98 ± 9. 93 )% those was lower compared with (21 ～30) score group( t = 2. 968, P ＜ 0. 01 ). In death group, APACHE Ⅱ score(28. 1 ± 6. 7 ) was higher than that in survival group ( 18. 8 ± 8. 4) ( t = 3. 030, P ＜0. 01 ), the early lactate clearance rate was lower ( t = 3. 619, P ＜ 0. 01 ). APACHE Ⅱ score correlated well with the mean level of the early lactate clearance rate ( r = - 0. 641, P ＜ 0. 01 ). Conclusion The lactate clearance rate was the good fator on evaluation of condition and prognosis in the critically ill patients.

Objective To study the relieving cough effect ofPuji-Xuanfei-Xiaodu decoction on acute bronchitis.Methods 87 patients with acute bronchitis of exterior cold and interior heat were randomly divided into two groups(44 cases in a therapy group and 43 cases in a control group).Puji-Xuanfei-Xiaodu decoction was given to the patients in the therapy group and acute bronchitis syrup was given to the control group, to observe the influence on symptoms of cough, sputum, fever and headache. The course of treatment was 7 days. Results On the reliving cough aspect, 28 cases (63.6%) showed cure effect and 13 cases (29.5%) effctive, the total effective rate was 93.2% in therapy group with 44 cases; while in control group (43 cases), 15 cases (43.9%) showed cure effect and 23 cases (53.5%) effctive, the total effective rate was 88.4%. The symptoms of cough, phlegm, fever, headache and aversion to wind were significantly decreased in two groups. In therapy group, the numbers of cure effect, remarkable effect and efficacy were 22 (50%) ,11(25%) and 8 (18.2%) cases respectively, the total effective rate was 93.19% while the numbers of cure effect, remarkable effect and efficacy were 14 (32.6%) ,8(18.6%) and 12 (27.9%) cases respectively, the total effective rate was 79.07% in control group. The total effective ratio of two groups had significant difference (P>0.05).ConclusionsPuji-Xuanfei-Xiaodu decoction is effective in relieving cough.

Objective To investigate the acute toxicity of intravenous isoflurane in Beagles.Methods Six healthy adult Beagles of both sexes aged 6-8 months weighing 6-8 kg were used in this study.Isoflurane injectio (120 mg/ml) in 30% hpid emulsion was injected intravenously. Femoral artery was cannulated for direct BP monitoring.ECG was continuously monitored.The maximal tolerance dose (MTD) and approximate lethal dose (ALD) were determined by up-and-down technique. The initial dose was 3.0 ml/kg. The dose was decreased/increased by 0.3 ml/kg if the previous animal died/survived.The survived dogs were observed for 2 weeks.Autopsy and histopathological examination were performed on all dead Beagles.Results The ALD and MTD of intravenous isoflurane were 252 and 216 mg/kg. Autopsy and histopathological examination did not show any abnormality.Conclusion Cardiopulmonary depression is the main manifestation of the acute toxicity of intravenous isoflurane in Beagles.

Objective To assess the validity of the Oxford classification for pediatric patients with primary IgA nephropathy (IgAN) and to analyze the correlations between clinical characteristics at time of biopsy and the Oxford classification,which identified four definitive histological features:mesangial hypereellularity,endocapillary proliferation,segmental sclerosis and tubular atrophy/interstitial fibrosis.Methods Clinical and pathological characteristics of 35 children with primary IgAN were analyzed.The scoring sheet was based on the Oxford classification of IgAN,and four pathological variables,namely mesangial hypercellularity (M),endocapillary proliferation (E),segmental sclerosis (S),and tubular atrophy/interstitial fibrosis (T) were assessed.A total of 35 children with IgAN were grouped according to the scores(M,E,S,T):the M0 and M1 group,E0 and E1 group,S0 and S1 group,T0 and T1/T2 group.These groups were compared in terms of estimated glomeralar filtration rate (eGFR),mean artery pressure (MAP) and proteinuria at time of biopsy.Results We found that the Oxford classification was significantly negatively correlated with eGFR (Pearson's correlation coefficient r =-0.48).However,the Oxford classification was shown to be positively associated with initial proteinuria per day(Pearson's correlation coefficient r =0.35).The M,E,S,T scores were strongly associated with proteinuria at biopsy (P ＜ 0.05),and the lesion S was not correlated with eGFR (P ＞ 0.05).The lesion T was significantly associated with eGFR (P =0.001) and MAP (P =0.03) at biopsy.Conclusion We confirmed that the Oxford classification of IgA nephropathy was valid for children.In addition,our study indicated that the four histological lesions M,E,S and T were significantly associated with clinical features.

Objective To observe the effect of recombinant human mannose-binding lectin ( rhMBL) on apoptosis of intestinal epithelial cells. Methods Fourteen C57 mice were randomly divided into two groups. The LPS mice received intra-peritoneal injection of LPS for indu-cing acute intestinal barrier injury. The expression of MBL protein and mRNA were observed by immuno-histochemisty and RT-PCR respec-tively. The apoptosis of intestinal epithelial cells was detected by tunel staining. In vitro, the Caco-2 cells were treated with 0、 5、10、20 μg/mLrhMBL, flow cytometry was used to detect the apoptosis rate of Caco-2 cells. The Bax and Bcl-2 protein and mRNA were detected by Western blotting and Real-Time PCR methods between various groups. MAPK signal proteins were detected by Western blotting method. Results The MBL and TUNEL staining were apparently increased under LPS condition. The apoptotic rates of Caco-2 cells which were trea-ted by different rhMBL concentrations after 48 h were increased in 20μg/mL rhMBL when compared with other groups. The apoptotic rates of Caco-2 cells which were treated by 20 μg/mL rhMBL after different time were increased in time-dependent manner. The results of Western blotting and Real-Time PCR assay showed that levels of Bax/Bcl-2 was increased in Caco-2 cells which were treated by 20μg/mL rhMBL af-ter 48h. Meanwhile, the phosphorylation of P38 and ERK were obviously increased in Caco-2 cells. The inhibitory effect of ERK and P38 in-hibitors were studied by Western blotting. Our current study showed that a block in P38 MAPK dependent cell death might contribute to the decrease in 20μg/mL MBL-mediated Caco-2 apoptosis. Conclusion High concentration of rhMBL may play an important role in the intesti-nal epithelial cell apoptosis, and P38 pathway may be involved in this regulation.