Objective: To investigate the effect of Atractyloside on sevoflurane postconditioning in focal cerebral ischemia.Methods: Fifty male SD rats were randomly assigned to five groups(n=10 each): control group(con),1.0 MAC group(sevo),1.0 MAC+ Atractyloside group(sevo+Atr),1.0 MAC+ vehicle group(sevo+vehicle)and Atractyloside group(Atr).All animals were subjected to the right middle cerebral artery occlusion(MCAO) for 120 min followed by reperfusion for 72 h.The animals in sevo groups were given 1.0 MAC sevoflurane inhalation from 20 min before to 10 min after reperfusion.The animals in Atr groups were given Atractyloside by ICV injection before sevoflurane postcondioning.The neurological deficit scores(NDS) were recorded at 24 h,48 h,and 72 h after reperfusion.Infarct volume percentage was determined after the last NDS assessment.Results: The infarct volume percentage ratio of sevo group(0.32?0.05) was significantly lower than that of con group(0.55?0.07)(P0.05).The infarct volume percentage ratio of sevo+Atr group and Atr group were significantly higher than that of Sevo group(P0.05).NDS of 24 h,48 h and 72 h after reperfusion in sevo group and sevo+vehicle were significantly higher than that in Con group.NDS of 24 h,48 h and 72 h after reperfusion in sevo+Atr group and Atr were significantly lower than that in Sevo group.Conclusion: Atractyloside abolished the protective effects of sevoflurane postconditioning on focal cerebral ischemia-reperfusion injury.

Objective:Ischemic cerebral injury is a common complication during perioperative period.We found that ischemic postconditioning and various non-ischemic postconditioning(e.g.inhalation anesthetics) could significantly attenuate ischemic cerebral injury.The present study was to investigate the effect of Cyclosporin A on sevoflurane postconditioning in focal cerebral ischemia.Methods:Fifty male SD rats were randomly assigned to five groups (n=10 each): control group(con),1.0MAC group(sevo),1.0MAC+ Cyclosporin A group(sevo+CsA),1.0MAC+ vehicle group(sevo+vehicle)and Cyclosporin A group(CsA).All animals were subjected to right middle cerebral artery occlusion(MCAO) for 120min followed by reperfusion for 72h.The animals in sevo groups were given 1.0MAC sevoflurane inhalation from 20min before to 10min after reperfusion.The animals in CsA groups were given CsA by ICV injection before sevoflurane postconditioning.The neurological deficit scores(NDS) were recorded at 24h,48h,and 72h after reperfusion.Infarct volume percentage was determined after the last NDS assessment.Results:The infarct volume percentage ratio of Sevo, Sevo+CsA,CsA and Sevo+vehicle groups were 0.31?0.04,0.25?0.04,0.30?0.03 and 0.33?0.05,respectively(P

Objective To investigate the efficacy and safety of intrathecal chemotherapy combined with concurrent radiotherapy in patients with leptomeningeal metastases from solid tumors.Methods The clinical and follow-up data of 29 patients with leptomeningeal metastases from malignant solid tumor who had intrathecal chemotherapy combined with concurrent radiotherapy were retrospectively analyzed.The treatment regimen was that 12.5-15.0 mg of methotrexate intrathecal injection once a week for 8 successive weeks combined with whole brain irradiation to a total dose of 40 Gy,20 fractions or with lumbosacral spinal canal irradiation to a total dose of 50 Gy,20-25 fractions.Results Nineteen patients completed the concurrent therapy and 19 patients were dead.There were 9 patients with complete remission of symptoms,11 with obvious improved symptoms,6 with alleviated symptoms,and 3 with no relief.All of the 29 patients,were followed up for 0.4-15.0 months.The median survival was 5 months.The 6-month survival rate was 48％ (11/23),excluding 4 patients with follow-up time less than 6 months and 2 patients noncancer deaths.The main toxicities were myelosuppression,methotrexate-related mucositis,chemical nerve root injuries,chemical meningitis,chronic neurotoxicity,seizures,and radiation-related mucositis.The incidence of severe toxicity was 17％ (5/29).Conclusions Combination of intrathecal methotrexate with concurrent radiotherapy can effectively alleviate the symptoms and improve the life quality of patients.The therapy with low incidence of severe toxicity and good tolerance has the trendency to prolong the median survival.

Aim To study the anti-inflammatory activity of the Channa argus bile.Methods The bile was isolated and purified by extraction and silica gel column chromatography.Then the compounds were identified by hydrogen and carbon spectra.The spleen lymphocytes proliferation assay and Lipopolysaccharide(LPS) induced mouse macrophage RAW264.7 releasing Nitrogen Monoxide(NO) experiment were used to evaluate the anti-inflammatory activity.Results Compound(C1) of sodium taurocholate and compound(C2) of sodium taurochenodeoxycholate were isolated by activity tracing.The cell relative viabilities of the two compounds on Concanavalin A(Con A) induced spleen lymphocytes proliferation assay were 65.9%±11.7% and 60.5%±9.4%, which were significantly different from the result of model group (P<0.01), respectively.The NO production of LPS-induced RAW264.7 release of NO was (16.4±1.9) μmol·L-1 and (15.5±1.7) μmol·L-1, which were significantly different from the result of model group(P<0.01).Conclusion Sodium taurocholate and sodium taurochenodeoxycholate from Channa argus perform the anti-inflammatory activities but have no cytotoxic effect on spleen lymphocytes and macrophage.

Objective To develop an HPLC method for the measurement of n-3 fatty acid index of serum cholesteryl esters.Methods Serum triglycerides were hydrolyzed with ethanolic sodium hydroxide and cholesteryl esters (CEs) were extracted with hexane.The extracted CEs were analyzed by reversed phase HPLC with a UV detection at 205 nm.Cholesteryl eicosapentaenoate and docosahexaenoate ( major n-3 fatty acid cholesteryl esters) were identified by liquid chromatography-tandem mass spectrometry and cholesterol in each CE fraction was measured.Peak areas of CEs were corrected for cholesterol and CE n-3 index was calculated using the corrected peak area and expressed as the percentage of n-3 fatty acid CEs in total CEs.Results The HPLC analysis can be finished in 6 minutes.Triglycerides which interfere with the determination of n-3 fatty acid index, were hydrolyzed with ethanolic sodium hydroxide (4 mol/L) in 30 seconds.The within-run and total CVs for CE n-3 index averaged 0.66% and 0.90%, respectively.CE n-3 indexes of 70 volunteers and 36 coronary heart disease patients apparently healthy subjects and patients with coronary heart disease in Beijing Hospital appeared to be positively skewed and leptokurtic distribution ( skewness = 1.25, kurtosis = 1.70 ).The median of n-3 indices were 0.98% ( 0.37% - 2.40% ).The logarithm of n-3 index appeared to be normal distribution and the average is 0.003 7% with standard deviations of 0.15.The distribution of n-3 indices of gender groups was similar with the total.The medians of females and males were 1.08% (0.60% -2.40%) and 0.95% (0.37% -2.11%) respectively, and the former were significantly higher than the latter( t = - 3.021, P = 0.003 ).Conclusion A new method for the measurement of n-3 index of serum cholesteryl esters by HPLC has been established.It is simple and precise and can be used in predicting cardiovascular diseases risks and monitoring dietary intake of n-3 fatty acids.

Objective:To study the association of air pollution with health service demand of the elderly and middle-age patients with cardiovascular and cerebrovascular diseases,and to provide a scientific ba-sis for development of environmental protection policy and health service policy of the Chinese govern-ment.Methods:This study included survey data on self-evaluated health,outpatient service demand and inpatient service demand of the patients with hypertension,heart disease and stroke in 62 cities of 1 7 provinces from China Health and Retirement Longitudinal Study(CHARLS)in 201 1 and 201 3,and com-bined it with the data on the annual concentrations of inhalable particulate matter(PM1 0 ),sulfur dioxide (SO2 )and nitrogen dioxide(NO2 )of those provinces and cities.Conditional Logistic regression was car-ried out to assess the possible effects of air pollutants on self-evaluated health and health service utiliza-tion.Results:The results showed that turning points existed in the effects of concentrations of NO2 and SO2 on the health service demand of the patients with hypertension,heart disease and stroke.The inpa-tient service demand of the hypertension patients increased with NO2 concentration when it was lower than 35.1 μg/m3 and decreased with NO2 concentration for higher value.Self-evaluated health of the patients with heart disease and stroke decreased with SO2 concentration when it was lower than 63.8 μg/m3 and increased with SO2 concentration for higher value.In addition,no evidence was found for the association between PM1 0 and health service demand.Conclusion:Air pollution may have effects on health service demand of the patients with hypertension,cardiovascular and cerebrovascular diseases,and different air pollutants at high or low concentration may have different health effects.

Objective To establish a method for measuring serum cotinine by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS) and provide an assay that can be applied to theevaluation of the level of smoke exposure and to the risk analysis of smoking related diseases.Methods Blood samples were collected from 94 apparently healthy subjects from October to December in 2010 and centrifuged,and the sera were separated.Serum samples were mixed with [ D3 ] -cotinine ( as the internal standard) and treated with acetonitriles to precipitate protein.After centrifugation,the supernatants were transferred and evaporated under a stream of nitrogen until dryness and reconstituted with mobile phase.Then the residuals were analyzed by LC/MS/MS system with multiple reaction monitor model; the concentration of cotinine were quantified by the isotope internal standard method and the stand curve was employed with a series of calibration.To estimate the precision of the method,five frozen serum pools were repeatedly analyzed in five runs,and every pool was analyzed in triplicate.In addition,the recovery rates were analyzed with the serum sample added with different levels of standard.The stability of cotinine in serum preserved at room temperature,4 ℃ and - 80 ℃,respectively.Finally,the levels of cotinine of 94 healthy subjects were measured to evaluate the distribution of cotinine with different smoke statuses.Results Serum cotinine measured by ID-LC/MS/MS was separated well with few interferences.The correlation coefficients between the peak area ratios and cotinine concentrations were higher than 0.9993.The values of within-run coefficients of variation (CV) of five frozen serum pools (0.68,48.42,94.34,250.95 and 287.04 μg/L) were 2.19％,0.78％,0.75％,0.65％ and 0.67％,respectively.The values of total CV were 4.71％,1.40％,1.98％,1.10％ and 1.03％,respectively.The limit of detection (LOD) and limit of quantitation ( LOQ ) were 0.013 and 0.050 μg/L,respectively.The analytical recoveries ranged from 99.22％ to 102.67％.The samples could maintain stability within 2 d at room temperature,7 d at 4 ℃ and 3 months at -80 ℃ resulting the accuracy of measurements from 99.28％ to 100.87％ and the CV＜5％.The levels of cotinine of 94 healthy subjects were measured and shown skewed and leptokurtic distribution.The concentrations of twenty smokers,fourteen former smokers and sixty non-smokers were 116.40 (63.17 -241.12),0.67 (0.15 - 0.95 ) and 0.22 (0.15 - 0.42 ) μg/L,respectively.Furthermore,the level of cotinine of former smokers (Z =-2.12,P ＜0.05) and smokers (Z =-6.67,P ＜0.001) were statistically higher than non-smokers.Conclusions An ID-LC/MS/MS method for serum cotinine detection has been established.It is hoped that the method will be applied to the assessment of smoke exposure and its association with the risks of smoking related diseases since it is simple,specific,precise,sensitive and accurate.

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.

Objective To develop a method for measuring serum total glycerol by isotope dilution (ID)liquid chromatography tandem mass spectrometry(LC/MS/MS).Methods Serum samples were mixed with [13C3]-glycerol(internal standard)and treated with alcoholic potassium hydroxide to hydrolyze serum glycerides to glycerol.The natural glycerol and the internal standard were benzoylated with the Schotten-Baumann reaction and the benzoates were analyzed by LC/MS/MS.Results The correlation coefficients between the peak area ratios and glycerol concentrations were 0.999 9 and higher.The withinrun coefficients of variation(CV)for serum total glycerol analysis averaged 0.94%(ranged 0.21%～2.62%)and the total CV 1.15%(0.62%～2.00%).Results on certified reference materials(SRM 909b Level Ⅰ and LevelⅡ;GBW 09146 and GBW 09147)showed an averaged bias of 0.20%(-0.20%～1.06%).Conclusions An ID-LC/MS/MS method for serum total glycerol has been developed.The method iS highly specific and precise and may be used as a candidate reference method for serum total glycerol measurements.

Objective To investigate the role of mucosal mast cells infiltration and degranulation with nerve growth factor (NGF)in development of visceral hypersensitivity in Sprague-Dawley (SD)rats. Methods The model of visceral hypersensitivity of irritable bowel syndrome (IBS)was established in 19 neonate SD rats with intestinal stimulation (rectalballon distention)on 8th,10th and 12th postnatal days. The other 19 neonate SD rats without colonic distention were assigned to the control group.After rats grew up (six to eight weeks old),the visceral sensitivity was tested by abdominal withdrawal reflex (AWR)in 10 rats of each group.Mast cell infiltration and degranulation were observed with toluidine blue staining in colon tissue slides.The NGF level of intestinal tissues was detected by enzyme-linked immunosorbent assay (ELISA)methods in the left nine rats of each group.The culture system of dorsal root ganglias (DRG)from the neonatal rats was set up.The changes of electrophysilogical characters of DRG stimulated with NGF (100 ng/mL)for four days were recorded with patch-clamp.Paired t test was performed for comparison between groups.Results The results of AWR indicated that neonatal colonic stimulation could significantly increase visceral sensitivity after growing up.Under 20,40 and 60 mmHg (1 mmHg=0.133 kPa)distention pressure,visceral sensitivity scores of visceral hypersensitivity rats and rats of control group were 1 .00±0.50 vs 1 .67 ±0.50,1 .89 ±0.31 vs 2.89 ±0.34 and 2.89 ±0.33 vs 3.89±0.33,the differences were statistically significant (t=-2.83,-6.00 and -6.00,all P <0.05 ). The results of master cells staining in tissue slides showed colonic master cells infiltration was obvious in rats with visceral hypersensitivity,and part of mast cells were degranulation.The result of ELISA demonstrated that NGF level of visceral hypersensitivity rats was significantly higher than that of control group ((11.07±3.06)pg/mg vs (2.38 ±1.88)pg/mg,t =-6.93,P <0.05).The results of electrophysilogical tests of primary cultured DRG indicated that compared with blank control growp,the action potential threshold of neuron in NGF 100 ng/mL group significantly decreased ((-18.0±2.1 )mV vs (-29.0 ± 2.5 )mV,t = 12.26,P <0.05)and discharge frequency increased ((5 .0± 1 .4 )/800 ms vs (12.0 ± 3.2)/800 ms,t=-8.40,P <0.05 ).Meanwhile,neuron voltage-gated K+ current density remarkably decreased,most were sustained delayed rectifier K+ current (I K )decreasing ((279.0 ±48.0)pA/pF vs (203.0±39.0)pA/pF,t=6.18,P <0.05).Conclusion Colonic stimulation in neonatal rats could cause intestinal master cells infiltration and degranulation,which induced changes of neuron electrophysilogical characters and resulted in visceral hypersensitivity after growing up.