Objective To evaluate the relationship between calmodulin protein kinase Ⅱ (CaMK Ⅱ) and levosimendan against arrhythmias induced by myocardial ischemia-reperfusion (I/R) in rats in vitro.Methods Thirty male Wistar rats,weighing 250-300 g,were randomly divided into 3 groups (n =10 each) using a random number table:control group (group C),I/R group and levosimendan group (group L).Their hearts were rapidly excised and perfused in a langendorff apparatus with K-H solution saturated with 95％ O2-5％ CO2 at 36.5-37.5 ℃.At 20 min of equilibration,the hearts were perfused with K-H solution for 60 min in group C.The hearts were subjected to 30 min of ischemia followed by 30 min reperfusion with K-H solution in group I/R.The hearts were subjected to 30 min of ischemia followed by 30 min reperfusion with K-H solution containing 300 nmol/L levosimendan in group L.Left ventricle developed pressure (LVDP),left ventricle end-diastolic pressure (LVEDP),+ dP/dt-dP/dtmax and heart rate (HR) were recorded immediately before ischemia and at 15 and 30 min of reperfusion.Arrhythmia was recorded during reperfusion and scored.Specimens were obtained from the apex of heart at 30 min of reperfusion for determination of the intracellular calcium concentration ([Ca2 +] i).Myocardial specimens were obtained from the left ventricle at 30 min of reperfusion to detect CaMK Ⅱ activity.Results Compared with group C,arrhythmia score,[Ca2+]i and CaMK [Ⅱ activity were significantly increased,and LVDP,+ dP/dtmax,-dP/dtmax and HR were decreased,and LVEDP was increased at 15 and 30 min of reperfusion in group I/R.Compared with group I/R,the number of ventricular premature beat,arrhythmia score,[Ca2+] i and CaMK Ⅱ activity were significantly decreased,and LVDP,+ dP/dtmax,-dP/dtmax and HR were increased,and LVEDP was decreased at 15 and 30 min of reperfusion in group L.Conclusion Inhibition of CaMK Ⅱ activity is involved in the mechanism by which levosimendan decreases the development of arrhythmias induced by myocardial I/R in rats.

Objective Endoscopy, 24-hour esophageal pH monitoring were used to objectively estimate the extent of gastroesophageal reflux and affecting factor after esophagectomy for cancer. Methods Endoscopy, 24 hour pH monitoring were performed in thirty-nine patients, including 21 undergoing esophagogastrostomy above the aortic arch (group A) and 18 below the aortic arch (group B). Results (1) DeMeeter score of gastroesophageal reflux in group A was significantly higher than that in group B in different postoperative period (P0.05). Conclusion Thoracic anastomosis in the lower thorax is more likely to be followed by gastroesophageal reflux and esophagitis. There is no significant change in extent of gastroesophageal reflux over time.

Objective To observe the change of serum smooth muscle myosin heavy chain (smMHC) level in the patients with aortic dissection (AD),and evaluate the effect of smMHC in the early diagnosis and prognosis of AD.Methods Forty-two patients with AD were selected as AD group,30 healthy subjects were selected as control group.Blood samples were collected at four time periods (within 3 h of onset,6 h,12 h,24 h),and serum smMHC level were measured by enzyme-linked immunosorbent assay.Results Serum smMHC level of AD group,which collected (within 3 h of onset,6 h,12 h) were significantly higher than that of control group [(88.6 ±21.7),(59.4 ± 18.7),(41.3 ± 10.7) ng/L vs.(17.2 ± 8.3) ng/L,P ＜ 0.01].There was no significant difference between the serum smMHC level of AD group and control group at 24 h after onset [(18.9 ±9.5) ng/L vs.(17.2 ±8.3) ng/L,P ＞ 0.05].Serum smMHC level of Stanford A type group (25 cases) was higher than that of Stanford B type group (17 cases) within 3 h of onset [(95.4 ± 17.8) ng/L vs.(78.5 ± 18.3) ng/L,P＜0.01],and there was no significant difference bewteen the two groups which collected at 6,12 h and 24 h after onset (P ＞ 0.05).Preoperative serum smMHC level was significantly higher than that after intracavitary isolation operation [(58.6 ± 15.9) ng/L vs.(30.1 ± 12.5) ng/L,P ＜ 0.01].Serum smMHC level decreased rapidly after the operation,and there was no significant difference between the two grougs when 12 h after operation [(18.7 ± 8.9) ng/L vs.(17.2 ± 8.3) ng/L,P ＞ 0.05].The serum smMHC level of the deaths (7 cases),which collected within 3 h of onset,6 h,12 h,was significantly higher than that of the survivors (35 cases) [(101.2 ± 20.7) ng/L vs.(86.1 ± 18.9) ng/L,(65.2 ± 16.7) ng/L vs.(58.2 ± 14.2) ng/L,(50.4 ± 10.8) ng/L vs.(39.5 ± 8.3) ng/L,P ＜ 0.05],and there was no significant difference at 24 h after onset (P ＞ 0.05).Detecting serum smMHC level within 3 h of onset,the area under the receiver operating characteristic curve was 0.913,with 51.7 ng/L as a diagnostic critical value,sensitivity and specificity respectively was 88.1％ (37/42) and 96.7％ (29/30).When detecting at 6 h after onset,the area under the curve was 0.865,with 38.5 ng/L as a diagnostic critical value,sensitivity and specificity respectively was 90.4％(38/42) and 90.0％ (27/30).Conclusions The level of serum smMHC in patients with AD increase rapidly after onset,and detecting serum smMHC level within 6 h of onset have important clinical significance in early diagnosis and prognosis of AD.

Objective:To evaluate the molecular diagnosis marker of papillary thyroid carcinoma (PTC),the relationship between lymphatic metastasis of central neck compartment PTC,and the opera-tion indication of prophylactic central neck dissection.Methods:We conducted a retrospective study, including 275 PTC patients and detected their BRAF mutation rates during 201 2 and 201 4 and explored the risk factors of the central node lymphatic metastasis by Logistic regression model.Results:Of the 275 PTC patients,224 (81 .5%)were female and 51 (1 8.5%)were male.BRAF mutational rates were 53.8% (1 48 /275)and lymphatic metastasis 57.8% (1 59 /275).Multivariate analysis showed calcifica-tion (ORadjusted =1 .47,95%CI:1 .1 0 -1 .98,P =0.01 ),tumor diameter (ORadjusted =1 .48,95%CI:1 .04 -2.30,P =0.048)and age (ORadjusted =1 .48,95%CI:1 .04 -2.30,P =0.048)were associa-ted with lymphatic metastasis.In stratified analysis,BRAF mutation (ORadjusted =3.1 9,95%CI:1 .1 8 -9.43,P =0.023 )in clear boarder group and BRAF mutation (ORadjusted =4.84,95% CI:1 .68 -1 3.84,P =0.003)in calcification group were more likely to have lymphatic metastases.Conclusion:Central neck metastasis takes up a high ratio in papillary thyroid cancer patients,BRAF mutation in pa-pillary thyroid carcinoma is a characteristic molecular event.Furthermore,patients with calcification un-der ultrasound detection,lower age group and longer tumor diameter are more susceptible to suffer central neck metastasis.Especially for stratified analysis,non-calcified BRAF mutation or BRAF mutation with clear border under ultrasound detection are more susceptible to suffer central neck metastasis,and radical prophylactic central neck dissection should be carried on for these patients.

Objective To evaluate the effects of sevoflurane preconditioning on zonula occludens-1 (ZO-1) during myocardial ischemia-reperfusion (I/R) in rats in vitro.Methods Adult male Wistar rats,weighing 250-300 g,were anesthetized with intraperitoneal pentobarbital 30 mg/kg and heparinized.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95％ O2-5％ CO2 at 37 ℃.Eighteen isolated rat hearts were randomly assigned into 3 groups (n =6 each) using a random number table:control group (group C),group I/R and sevoflurane preconditioning group (group S).At 10 min of equilibration,the hearts were perfused with K-H solution for 110 min in group C,the hearts were perfused with K-H solution for 20 min,and then subjected to 30 min of ischemia followed by 60 min of reperfusion in group I/R,and the hearts were perfused with K-H solution saturated with 3％ sevoflurane for 15 min followed by 5 min washout,and then subjected to 30 min of ischemia followed by 60 min of reperfusion in group S.At the end of equilibration,immediately before ischemia,and at 15 and 60 min of reperfusion (T1,2),HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP),+ dp/dtmax and-dp/dtmax were recorded.The development of arrhythmias was recorded during reperfusion and scored.At 60 min of reperfusion,myocardial specimens were obtained from the apex of heart for determination of the expression of ZO-1 in myocardial tissues (by Western blot) and for observation of distribution of ZO-1 and connexin43 (Cx43) (by immunofluorescence).Results Compared with group C,HR,LVDP,+ dp/dtmax and-dp/dtmax were significantly decreased and LVEDP was increased at 15 and 60 min of reperfusion,scores of arrhythmia was increased,and ZO-1 expression was down-regulated in I/R group.Compared with group I/R,HR,LVDP,+ dp/dtmax and-dp/dtmax were significantly increased and LVEDP was decreased at 15 and 60 min of reperfusion,arrhythmia was decreased,and ZO-1 expression was up-regulated in group S.ZO-1 and Cx43 were co-localized at the intercalated disk.ZO-1 was redistributed in the lateralization of the membrane and co-localized with Cx43 in group I/R.The incidence of ZO-1 lateralization was significantly decreased in group S.Conclusion The mechanism by which sevoflurane preconditioning decreases reperfusion arrhythmia is related to inhibition of down-regulation of expression and redistribution of ZO-1 and inhibition of redistribution of Cx43 in rats.

Objective To evaluate the effects of sevoflurane preconditioning on wnt/glycogen synthase kinase-3 beta (GSK-3β)/β-catenin signaling pathway during myocardial ischemia-reperfusion (I/R) injury in rats in vitro.Methods Ault male Wistar rats,weighing 220-280 g,were heparinized and anesthetized with intraperitoneal 3％ pentobarbital 30 mg/kg.Their hearts were rapidly excised and perfused in a langendorff apparatus with oxygenated (95％ O2-5％ CO2) K-H solution at 37 ℃.After 15 min of equilibration,36 isolated hearts were randomly divided into 3 groups (n=12 each) using a random number table:sham operation group (group S),group I/R and sevoflurane preconditioning group (group SP).After 30 min of equilibration,the hearts were continuously perfused for 150 min in group S.The isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion.In SP group,the hearts were perfused for 15 min with K-H solution containing 2.4％ sevoflurane,followed by 5 min washout before reperfusion.At the end of equilibration and 30 min of reperfusion,HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and ± dp/dtmax were recorded.The severity of arrhythmias was assessed during reperfusion.At 60 min of reperfusion,3 hearts in each group were chosen for measurement of expression of wnt3a,phosphor-GSK-3β (p-GSK-3β) and β-catenin (by Western blot).At 120 min of reperfusion,6 hearts in each group were chosen for determination of myocardial infarct size by TTC staining.Results Compared with group S,HR,LVDP,+dp/dtmax and -dp/dtmax were significantly decreased,and LVEDP was increased at 30 min of reperfusion,arrhythmia scores and the percentage of myocardial infarct size were increased,and the expression of wnt3a,p-GSK-3β and β-catenin was down-regulated in I/R group.Compared with group I/R,HR,LVDP,+dp/dtmax and-dp/dtmax were significantly increased,and LVEDP was decreased at 30 min of reperfusion,arrhythmia scores and the percentage of myocardial infarct size were decreased,and the expression of wnt3a,p-GSK-3β and β-catenin was up-regulated in group SP.Conclusion Sevoflurane preconditioning attenuates myocardial I/R injury by activating wnt/GSK-3β/β-catenin signaling pathway in isolated rat hearts.

0.05).Conclusions:Concurrent chemoradiotherapy can be well tolerated even though the acute side-effects less than grade 2 were higher in concurrent chemoradiotherapy than other group.Immediate response was very encouraging in the concurrent group.There was no advantage in terms of survival rate in the concurrent group compared to the sequential group.

Objective To investigate the role of phosphatidyl-inositol 3-kinase-Akt (PI3k-Akt) signal pathway in the attenuation of ischemia-reperfusion (I/R) injury by sevoflurane preconditioning in isolated rat hearts. Methods Ninety-six adult male SD rats weighing 220-280 g were randomly divided into 6 groups ( n = 16 each): sham operation group (group S); I/R group; sevoflurane preconditioning group (group SP); wortmannin group (group W); dimethyl sulfoxide (DMSO) group (group D) and sevoflurane preconditioning + wortmannin group (group SW) . Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%C02 at 37 ℃ . The hearts were continuously perfused for 180 min in group S. After 15 min of equilibration, the isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion in SP, W, D and SW groups. Croups SP, W, D and SW received 10 min of perfusion with K-H solution containing 2. 4% sevoflurane, 100 nmol/L wortmannin, 20 μmol/L DMSO, and 2.4% sevoflurane + 100 nmol/L wortmannin, respectively, followed by 5 min washout before I/R. Eight hearts in each group were selected and HR, left ventricular end-diabetic pressure (LVEDP), left ventricular developed pressure (LVDP), and ± dp/dtmax were recorded at the end of equilibration and at 15 min of reperfusion, Myocardial tissues were obtained at 15 min of reperfusion for determination of apoptosis (by TUNEL) and phosphorylated Akt (p-Akt) expression (by Western blot) . Another 8 hearts were selected at 120 min of reperfusion for determination of myocardial infarct size by TTC staining. Result Compared with group S, LVDP and ± dp/dt,^ were significantly decreased and LVEDP was significantly increased in groups I/R, SP, W, D and SW, and myocardial p-Akt expression was up-regulated in groups I/R, SP and D ( P ＜ 0.05). Compared with group I/R, LVDP and ± dp/dtmax were significantly increased, LVEDP and apoptosis index were significantly decreased, myocardial p-Akt expression was up-regulated, and myocardial infarct size was significantly reduced in group SP (P ＜0.05) . Conclusion Activation of PI3K-Akt signal pathway is involved in the attenuation of I/R injury by sevoflurane reconditioning in isolated rat hearts.

Objective To evaluate the effect of sevoflurane postconditioning on the expression of Pim-1 kinase during myocardial ischemia-reperfusion (I/R) in rats.Methods Male Sprague-Dawley rats,weighing 250-300 g,were used in this study.After the animals were anesthetized,their hearts were immediately removed and retrogradely perfused with an oxygenated K-H solution at 37 ℃ in a Langendorff apparatus.Thirty-six isolated rat hearts were assigned into 3 groups (n=12 each) using a random number table:control group (group C),group I/R and sevoflurane postconditioning group (group SP).The hearts were subjected to ischemia for 30 min followed by 120 min of reperfusion to establish the model of myocardial I/R injury.In group SP,the hearts were perfused with K-H solution saturated with 3％ sevoflurane for 15 min starting from the beginning of reperfusion.Heart rate (HR),left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and the maximum rate of increase and decrease of left ventricular pressure (±dp/dtmax)were recorded at the end of equilibration and 30,60,90and 120 min of reperfusion.Myocardial tissues were obtained at T2 for determination of the expression of Pim-1 kinase,Bcl-2 and cytochrome c (Cyt c) in cytoplasm and mitochondria by Western blot.The hearts were selected at T4 for determination of myocardial infarct size (by TTC staining) and for examination of mitochondrial ultrastructure (with transmission electron microscope).Results Compared with group C,HR,LVDP and ±dp/dtmax were significantly decreased,and LVEDP was increased at T1-4,the myocardial infarct size was enlarged,the expression of Pim-1 and Bcl-2 in cytoplasm and mitochondria was down-regulated,the expression of Cyt c in cytoplasm was up-regulated,and the expression of Cyt c in mitochondria was down-regulated in group I/R (P＜0.05).Compared with group I/R,HR,LVDP and ±dp/ dtmax were significantly increased,and LVEDP was decreased at T1-4,the myocardial infarct size was decreased,the expression of Pim-1 kinase and Bcl-2 in cytoplasm and mitochondria was up-regulated,the expression of Cyt c in cytoplasm was down-regulated,and the expression of Cyt c in mitochondria was upregulated in group SP (P＜0.05).The damage to mitochondria was significantly attenuated in group SP as compared with group I/R,and was aggravated as compared with group C.Conclusion The mechanism by which sevoflurane postconditioning attenuates myocardial I/R may be related to up-regulation of Pim-1 kinase expression in rats.

Objective To investigate the effect of sevoflurane postconditioning on mitochondrial connexin 43 (Cx43) during myocardial ischemia-reperfusion (I/R) in isolated rat hearts and the role of mitochondrial ATP-sensitive potassium (mito-KATP) channels in it.Methods Forty-five adult male SpragueDawley rats,weighing 200-250 g,were used in the study.Their hearts were excised and retrogradely perfused with K-H solution in a Langendorff apparatus.The 45 isolated hearts were assigned into 5 groups (n =9 each) using a random number table:control group (group C),I/R group,sevoflurane postconditioning group (group Sev),and sevoflurane postconditioning plus 5-hydroxydecanoate (5-HD,a specific mitoKATp channel blocker) group (group Sev+5-HD) and I/R plus 5-HD group (group I/R+5-HD).The hearts were subjected to 20 main of global ischemia followed by 90 min of reperfusion to establish the model of myocardial I/R injury.From the beginning of reperfusion,the hearts were perfused with K-H solution saturated with 3％ sevoflurane for 15 min in group Sev,with K-H solution saturated with 3％ sevoflurane and containing 100 μ mol/L 5-HD in group Sev+5-HD,and with K-H solution containing 100 μ mol/L 5-HD in group 5-HD.The heart rate (HR),left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and the maximum rate of increase and decrease of ventricular pressure (±dp/dtmax) were recorded at the end of equilibration (T1) and 30 and 60 min of reperfusion (T2,3).At 90 min of reperfusion,the myocardial infarct size was measured by TTC staining,and the expression of total Cx43 (tCx43)and phosphorylated Cx43 (p-Cx43) in mitochondria was determined by Western blot analysis.The percentage of myocardial infarct size and p-Cx43/tCx43 ratio were calculated.Results Compared with group C,the HR,LVDP and ±dp/dtmax were significantly decreased,and the LVEDP was increased at T2,3,and the percentage of myocardial infarct size was increased in the other 4 groups,the expression of mitochondrial tCx43 and p-Cx43 was significantly down-regulated in I/R,Sev+5-HD and 5-HD groups (P＜0.05),and no significant change was found in the expression of mitochondrial tCx43 and p-Cx43 in group Sev (P＞0.05).Compared with group I/R,the HR,LVDP and ±dp/dtmax were significantly increased,and the LVEDP was decreased at T2,3,the percentage of myocardial infarct size was decreased,and the expression of mitochondrial tCx43 and p-Cx43 was up-regulated in group Sev (P＜0.05),and no significant change was found in the parameters mentioned above in Sev+5-HD and 5-HD groups (P＞0.05).Compared with group Sev,the HR,LVDP and ±dp/dt were significantly decreased,and the LVEDP was increased at T2,3,the percentage of myocardial infarct size was increased,and the expression of mitochondrial tCx43 and p-Cx43 was down-regulated in group Sev+5-HD (P＜0.05).There was no significant change in the pCx43/tCx43 ratio between the five groups (P＞0.05).Conclusion The mechanism by which sevoflurane postconditioning attenuates myocardial I/R injury may be related to induction of mito-KATP channel opening and up-regulation of the expression of mitochondrial Cx43 in cardiomyocytes of rats.