Twelve patients with latent Keshan disease in Xunyi County,Shaanxi Province were electron-microscopically observed with endomyocardial biopsy. Seven of them were males, 5 females; their ages ranged from 14 to 40 years old. They had suffered from the disease for 4 to 5 years in an average. The samples taken from their heart ventricles were prepared in routine,some were prepared with the modified lanthanum preparation according to Hoffstein's method to study the membrane permeability of the patients' myocardium at the same time,In almost all the samples, most of sarcoplasmic reticula,transverse tubule system and intercalated disc spaces are dilated. In some samples prepared with the lanthanum preparation, the electronic density particles are found in the intercalated disc spaces and scarcely in the cytoplasm of myocardium. Mitochondria often show changes both of proliferation and of degeneration simutaneously. Sometimes, minimal myolytic changes of filaments also can be seen, but there are no any necrotic or myolytic foci of myocardium found in the samples at all. Some nuclei are enlarged and twised,in them some pseudoinclusions also can be occasionally seen. Myofibrils vary their thickness. At the same time, they also have some abnormal branches. More than normal amount of fat droplets and glycogen granules deposite in the myocardial cells. Besides,there are lots of lipofusicin granules accumulated around the nuclei in the cytoplasm.

Objective To establish a method to isolate,culture and identify cardiomyocytes from neonatal mouse,estimate the cardiomyocytes survival rate,and identify the purity of cardiomyocytes.Methods Neonatal C5 7 mouse heart(within 3 days)was di-gested using trypsin and collagenase.Cardiomyocytes survival rate was estimated by trypan blue staining,and cardiomyocytes purity was identified byα-actin immunofluorescence staining.Results Mouse cardiomyocytes could be successfully isolated and cultured using neonatal mouse within 3 days.Trypan blue staining showed cardiomyocytes survival rate was>95%,and cardiomyocytes pu-rity was more than 95% demonstrated byα-actin immunofluorescence staining.Conclusion Under the strict experimental condi-tions,mouse cardiomyocytes can be successfully isolated and cultured with high survival rate and high purity.

Objective To investigate the effect of preconditioning with mitochondrial ATP-sensitive potassium channel opener on the mitochondrial functions.Methods Thirty SD rats were used.Isoproterenol was used to induce myocardial ischemic injury in 20 rats,ten of which were pretreated with diazoxide.Rhodamine123(Rh123)was used as fluorescent prober to measure mitochondrial membrane potential.The activities of mitochondrial Na+-K+-ATPase and Ca2+-ATPase were detected.The mitochondrial respiratory parameters were recorded with a Clark electrode.The effect of diazoxide on mitochondrial membrane potential and mitochondrial respiration was investigated.Results Compared with the controls,the levels of state 3(ST3),respiratory control rate(RCR),mitochondrial membrane potential content and the activities of mitochondrial ATPase were decreased in the rats that received isoproterenol,while diazoxide pretreatment alleviated the changes of ST3,RCR,mitochondrial membrane potential and the activities of mitochondrial ATPase.Conclusion Preconditioning with diazoxide enhances the activities of mitochondrial ATPase and protects the myocardial mitochondrial function after isoproterenol induced myocardial ischemic injury.

AIM To study the effect of ketamine on proliferation,cell cycle in the cultured rat neural stem cells. METHODS The growth inhibition of ketamine on neural stem cell was evaluated by an MTT assay. The effect of ketamine on cell cycle was measured by flow cytometry. RESULTS Ketamine inhibited the growth of cultured rat neural stem cells. Flow cytometry analysis showed that G 0/G 1 phase rate was increased but S phase rate was decreased. CONCLUSION Ketamine can inhibit proliferation of cultured rat neural stem cells,and this inhibitition is associated with cell cycle block.

AIM: To investigate the effects of mitochondrial ATP sensitive potassium (MitoK_ ATP) channel opening on translocation of protein kinase C epsilon (PKC?) and the relationship between the translocation of PKC? and the production of reactive oxygen species. METHODS: The expression of PKC? in cultured adult rat ventricular myocytes was investigated with immunofluorescence and Western blotting techniques. RESULTS: (1) Diazoxide, a selective MitoK_ ATP channel opener, caused a significant translocation to myofibrillar-like structures in cultured adult rat ventricular myocytes. (2) The N-2-mercaptopropionylglycine (MPG), a free radical scavenger, partly inhibited the translocation of PKC? caused by diazoxide. (3) Chelerythrine, a selective protein kinase C (PKC) inhibitor, completely blocked the translocation of PKC? caused by diazoxide. CONCLUSION: Opening of MitoK_ ATP channel might activate PKC? and make it translocation to myofibrillar-like structures. PKC? activation occurs downstream of MitoK_ ATP channel, and might be caused by production of reactive oxygen species after opening of MitoK_ ATP channel.

Objective To analyze the clinical experience of double lumen endobronchial tube intubation in 41 cases of whole lung lavage .Methods Forty‐one patients with whole lung lavage in our hospital from February 2010 to February 2015 were retro‐spectively analyzed .The effect of bronchoscopy and auscultation location method were explored after double‐lumen endobronchial intubation in whole‐lung lavage .Results Among 41 cases ,39 cases were successfully located by using the auscultation location ,and other 2 cases were successfully positioned by using the bronchoscopy position after repeatedly auscultation location method resulting in lung isolation failure .The catheter depth in 19 cases was adjusted after using bronchoscopy ,and then double lung was isolated well and lavage was successfully conducted .Conclusion The prerequisite for success of whole lung lavage is accurate position of double‐lumen endobronchial tube .Bronchoscopy look positioning has become the gold standard of double‐lumen endobronchial tube position .

Objective To compare and analyze the clinical application of lumbar plexus combined with sciatic nerve block and spinal anesthesia for elderly patients with knee joint surgery.Methods A total of 77 elderly patients with ASAⅠ ~Ⅲ undergoing single knee re-placement surgery were randomly divided into combined group which recieved lumbar plexus combined with sciatic nerve block and spinal an-esthesia group.The baseline values,blood pressure and heart rate at multiple time points,the block area and duration,the volume of intraoper-ative fluid,and other indexes of adverse reaction were observed.Results The MAP,SBP and DBP in the spinal anesthesia group after the op-eration have changed significantly at the time of T1,T2 and T3.The operating of anesthesia in the combined group was shorter than that of spi-nal anesthesia group.The rate of adverse reactions in combined group was significantly lower than that inspinal anesthesia group.Conclusion The spinal anesthesia can be satisfied for operation requirements,but it will cause the unstable circulation and varied adverse reactions.Lum-bar plexus combined with sciatic nerve block is safe and effective with less adverse reactions,less disturbance of hemodynamics,which is much better for the old or the patients with coagulation abnormalities combined heart and lung disease.

Objective To evaluate the efficiency of dezocine combined with sufentanil on postoertive analgesia for spinal deformity sur-gery.Methods Divided the 90 patients who were admitted into our hospital from January 2013 to September 2015 into three groups by ran-dom single blind method,namely the dezocine group,the sufentanil group and the combined group,with 30 cases in each group.All the pa-tients underwnet propofol and sevoflurane static absorption compound anesthesia,and they were given continuous intravenous analgesia with different drugs after the surgery.Their visual analogue scale (VAS)score,sedation scale (SS)score,adverse reaction,total PCIA times,ef-fective PCIA times,respiratory rate and arterial blood gas were measured at 2,6,24,48 hours after operation.Results The VAS score of the combined group was lower than that of the dezocine group (P <0.05).The combined group was significantly superior to the dezocine group and the sufentanil group in SS score and adverse reactions.At 24 and 48 hours after surgery,SaO2 and PaO2 in the combined group were higher than those in sufentanil group (P <0.05).PaCO2 in the combined group was lower than that in the dezocine group and the sufentanil group(P <0.05).Conclusion Dezocine combined with sufentanil is a more efficiency way with less adverse reaction on postoperative anal-gesia for spinal deformity surgery,and it is an ideal way of analgesia.

AIM: To analyze and identify the phosphoproteins associated with diazoxide preconditioning. METHODS: Proteomics technique was used to investigate the changes of phosphoprotein after diazoxide preconditioning. Adult rat ventricular myocytes were pretreated in the presence and absence of 200 ?mol/L diazoxide for 10 min. Phosphoproteins prepared and enriched respectively from control and diazoxide pretreated groups were then separated by two-dimensional (2D) gel electrophoresis and stained with sliver staining kit. Phosphoproteins of interest were further identified by mass spectrometry. RESULTS: Associated with diazoxide preconditioning, the proteins of chaperonin containing TCP-1 and hypothetical protein XP_346548 were phosphorylated significantly. The proteins of 94 kD glucose-regulated protein, calpactin I heavy chain and ferritin were dephosphorylated markedly (P

Objective To explore the effects of sedationand hemodynamics on different ages patients with the same concentration of dexmedetomidine under general anesthesia.Methods A total of 264 patients (ASAⅠ-Ⅱ)with orthopaedic surgery under general anesthesia in our hospital from April 2013 to May 2015 were divided into 3 groups by age,the young group (group Y,n =76),middle age group (group M, n =107),and old age group (group O,n =81 ).Fifteen minutes before anesthesia,patients were infused dexmedetomidine with 1 μg/kg, maintain the concentration of 0.5 μg·kg-1 ·h-1 and stop at 30 minutes before surgery finished.The SBP、DBP、BIS、HR before anesthesia (T1),pump injection start(T2),tracheal intubation(T3),1 minute after intubation(T4),5 minutes after skin incision(T5),endotracheal ex-tubation(T6)were observed.The dosage of propofoland remifentanil in anesthesia,duration from stop infusion to endotracheal extubation, Ramsay score and adverse reactions 5 minutes after PACU also need to be recorded.Results The level of SBP and DBP were significantly increased at T2,T3 in all groups.Compared with group O,both group Y and group M increased significantly,the difference was statistically significant(P <0.05).The level of SBP and DBP were significantly decreased at T5 in all groups,the difference was statistically significant (P <0.05).There was no significant difference in 3 groups at T4-T6(P >0.05).Compared with T1,the level of HR and BIS were signifi-cantly decreased at T2-T5,the difference was statistically significant(P <0.05).The dosage of propofol and remifentanil in group Y and group Mwas more than that of group O.The extubation time was significantly shorter and the Ramsay score was significantly less than those of the group O,the difference was statistical significance(P <0.05).SAS scores among the three groups was not significant difference (P >0.05). There was no significant difference in the total adverse reaction between group Y and group M(P >0.05),but it was significantly lower than that of group O(P <0.05).Conclusion Dexmedetomidine has good sedative effect in all groups,but older group have more adverse reac-tions and wake up time is extended.The concentratiuon of dexmedetomidine should be adjusted according to the age of patients.