Objective To investigate the dynamic changes of ATP content in rat cerebral cortex after transient ischemia followed by reperfusion and the relationship between the change of energy and the recovery of neural function.Methods The rats were subjected to 10 min of middle cerebral artery occlusion (MCAO). At the time point of 0 h, 1 h, 3 h, 6 h, 12 h, 24 h and 72 h after reperfusion, ATP contents of frontal and parietal cortex were measured by capillary zone electrophoresis.Results At the end of 10 min ischemia, ATP content fell dramatically to less than 20% of the control level. After reperfusion, ATP content recovered gradually. After 1 h, 3 h, 6 h and 12 h of reperfusion, ATP content returned to 70.5%, 65.7%, 84.8% and 86.9% of the control level ( P=0.052, 0.030, 0.332 and 0.491). From 24 h on until 72 h after reperfusion, ATP content decreased again, reaching half of the control level ( P=0.003 and P=0.023). After 10 min ischemia, limb function recovered gradually and completely at last. From 24 h on until 72 h after reperfusion, unwillingness of action and eating was found.Conclusions The recovery of cellular energy system function is delayed even though the reperfusion is in time after transient cerebral ischemia. Furthermore, secondary failure of cellular energy system function occurrs with the reperfusion proceeding. These phenomena are probably responsible for the delayed recovery of neural function after cerebral ischemia in spite of reperfusion.

Objective To evaluate the effects of nucleoside/nucleotide analogue treatment on immunoglobulin and complement in patients with chronic hepatitis B (CHB).MethodsA total of 157 CHB patients were recruited and divided into CHB group,liver cirrhosis (LC) group and severe hepatitis B (SHB) group.There were 50 patients who received oral antiviral treatment (lamivudine 100 mg/d,or entecavir 0.5 mg/d,or telbivudine 600 mg/d).Serum levels of complement 3 and 4 (C3,C4),C-reaction protein (CRP),hemolytic complement (CH50),immunoglobulin G,M,A (IgG,IgM,IgA),hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) were detected by enzyme-linked immunosorbent assay (ELISA) or immunoturbidimetry.Hepatitis B virus (HBV) DNA was quantified by real-time polymerase chain reaction (RT-PCR) before and 1,2,3 and 4 weeks after nucleoside antiviral therapy.Comparison of means was done by t test and Mann-Whitney test.The correlation was analyzed by Pearson correlation coefficient test.ResultsSerum IgA and IgM levels of SHB and LC patients were significantly higher than those of CHB patients (P＜0.01).Levels of C3,C4,CH50 and CRP were significantly different among three groups.Levels of C3,IgM,IgG and HBV DNA in HBeAg positive patients were significantly different from those in HBeAg negative patients.There was a statistically significant difference of IgA,IgM,C3 and CH50 levels between patients with high HBV DNA level and low HBV DNA level in HBeAg-positive patients.While in the HBeAg-negative patients,only the IgA level was significantly different with HBV DNA levels.After anti-viral treatment,immunoglobulin and HBV DNA levels were all decreased in three groups,while the serum complement level was increased compared to baseline,and the differences became significant at week 4 of treatment. HBV DNA level was negatively correlated with C3 (r=-0.78,P=0.021) and HBeAg titer was positively correlated with C3 (r=0.87,P=0.015).ConclusionsThe immunoglobulin,CRP,C3,C4,and C H50 could reflect the inflammatory activity in liver.The changes of C3 level can predict the efficacy of antiviral therapy.

An analysis of the fundamental concepts for public hospital reform further defined public hospitals and other hospitals,business and non-business in nature,public benefits and welfare,as well as public benefits and profit-oriented hospitals.These distinctions can help reformers of public hospitals clarify their concepts,make clear the purpose and direction of the public hospitals reform,so as to speed up the reform with theory support.

This paper studied the development stages in terms of the public nature of public hospitals in China and analyzed the causes for their weakened public benefits.It proposed the principle of government guidance,achieving government purchasing of healthcare services as the roadmap,as well as building and improving the service purchasing system,performance appraisal system,and payment review system along with supervision system.These measures are designed to make public hospitals in their public benefit nature with scientific system design,powerful institutional guarantee,and efficient execution strategies.

Objective:To transfect genes using ultrasound targeted microbubble destruction (UTMD) techniques and observe the effects of RNA interference on cervical cancer (HeLa) cell line in silencing survivin gene and inducing apoptosis. Methods: Recombinant expression plasmid of short hairpin RNA (shRNA) targeting survivin gene was constructed. It was co-treated with microbubbles and transfected to cultured HeLa cells followed by exposure to ultrasound (P+UTMD group). Moreover, blank control group (C), plasmid group (P), ultrasound exposure group (US), plasmid and ultrasound exposure group (P+US), plasmid+ Lipofectamine group (P+L) were used as controls, respectively. Transfection efficacy was evaluated by observing the red fluorescence in the cells by fluorescent microscopy and flow cytometry(FCM). Ultrasound intensity and exposure time were optimized. Cell apoptosis was investigated using flow cytometry analysis, Hoechst staining, and DNA ladder method. Expression of survivin mRNA was assessed by RT-PCR. Results: Restrictive enzyme digestion and sequencing analysis verified that the recombinant plasmid was successfully constructed. UTMD significantly increased gene transfection efficacy in cultured HeLa cells (P<0.01). Gene transfer was the most prominent at ultrasound intensity of 1.0 W/cm2 and exposure time of 3 min (P<0.01). RT-PCR showed that the expression of survivin mRNA in P+UTMD group was inhibited by (83.33±2.73)%. The differences were significant compared with any other groups (P<0.01). FCM analysis showed that the apoptosis ratio in P+UTMD group was significantly increased as compared with other groups (P<0.01). Hoechst staining and DNA ladder showed that apparent apoptosis and DNA ladder were detected only in P+UTMD and P+L groups. Conclusions:UTMD effectively enhances the transfection efficacy of expression plasmid. It is a novel and effective non-viral gene transfer system and has promising foreground. UTMD mediates RNA interference silenced survivin gene and induces significant cell apoptosis, which provides a new method for tumor research and gene therapy.

Nucleotide-binding oligomerization domain protein 2 (NOD2) involves in host immune responses to pathogens and regulates natural immunity and specific immunity by identifying the peptidoglycan of bacterial cell walls and cleavage product muramyl dipeptide.As a newly discovered intracellular pattern recognition receptor,NOD2 plays important roles in the development of primary hepatocellular carcinoma by gene mutate,inducing liver inflammatory reaction and activating relevant signaling pathways.

Objective To analyze the curative effect of traditional Chinese medicine(TCM)combined with early enteral nutrition(EN)for treatment of patients with severe acute pancreatitis(SAP). Methods 70 SAP patients were randomly divided into TCM plus EN group(36 cases)and conventional therapy group(34 cases). Both groups received routine treatment. Additionally,TCM+EN group received early EN and TCM decoction treatment〔the ingredients of decoction including radix bupleuri,radix paeoniae alba,radix scutellariae,fructus aurantii immaturus, magnolia bark,raw rhubarb(rhubarb was added at last during cooking the decoction)and natrii sulfas exsiccatus (dissolved in water to be administered)each 10 g,the decoction was concentrated to 150 mL and then administered via a stomach tube to the patient,afterwards the tube was clipped for 2.5-3 hours,twice a day,4-7 days constituting a therapeutic course〕. After treatment,the time for patients' symptoms improvement,the situation of intestinal recovery, the length of stay in hospital,the total medical cost,serum C-reactive protein(CRP),aspartate aminotransferase (AST), lactate dehydrogenase (LDH), amylase (AMY), acute physiology and chronic health evaluationⅡ(APACHEⅡ)score and complications,intensive care unit(ICU)transfer rate and case fatality rate in two groups were observed. Results The time for symptoms improvement of abdominal tenderness(day:1.68±1.01 vs. 3.89±1.07), abdominal distension(day:2.17±1.48 vs. 4.24±3.23),abdominal pain(day:3.12±1.14 vs. 4.94±3.21)and the intestinal recovery of exhaust defecation time(day:3.48±0.92 vs. 5.32±3.30)of SAP patients after treatment in the TCM+EN group were faster significantly than those in the conventional therapy group(all P<0.05). The length of stay in hospital(day:15.50±1.75 vs. 19.35±1.69)and total cost(wan yuan:1.812±0.424 vs. 3.292±1.081) of TCM+EN group were less than those of conventional therapy group(P<0.05 or P<0.01). After treatment,the levels of serum CRP,AST,LDH,AMY,APACHEⅡscore in TCM+EN group and conventional therapy group were all lower than those before treatment,and on day 10,the degree of descent was more prominent in TCM+EN group〔CRP(mg/L):98.972±43.384 vs. 122.392±71.621,AST(U/L):75.952±55.668 vs. 126.391±47.431, LDH (μmol?s-1?L-1):1.48±0.21 vs. 2.61±1.46,AMY(U/L):146.362±58.792 vs. 226.392±37.692,APACHE Ⅱscores:6.978±3.352 vs. 13.652±7.621,P<0.05 or P<0.01〕. There was no death in TCM+EN group,while in the conventional therapy group,there was 1 case dead. ICU transfer rate in TCM+EN group was less than that in the conventional therapy group(2.78% vs. 11.76%),but there was no statistical significant difference between the two groups(χ2=0.99,P>0.05). Among the 70 patients with SAP,the cause of the disease due to gallstone accounted for 55.72%,hyperlipidemia for 37.14%,alcoholic for 4.28%and other 2.86%. Conclusion The use of TCM combined with early EN for treatment of patients with SAP can enhance the curative effect of SAP,reduce the hospitalization time and the total cost of patients,and decrease complications and mortality,that is conducive to the faster recovery of patients.

ObjectiveTo study the efficacy and safety of early endoscopic retrograde cholangiopancreatography(ERCP) and endoscopic sphincterotomy (EST) for patients with acute biliary pancreatitis.Methods420 patients who were admitted to our hospital for management of acute biliary pancreatitis were divided into early endoscopic therapy group (n=218 patients) and conservative therapy group (n=202 patients).The durations for complete disappearance of abdominal pain,decrease in serum and urine amylase levels to normal,liver function recovery time,white blood cell recovery time,and the mean duration and costs of hospitalizations were analyzed.ResultsIn the ERCP group,all patients received EST.The stones in 172 patients with choledocholithiasis were removed with dormia baskets or balloon catheters.In 20 patients with severe acute biliary pancreatitis,endoscopic pancreatic duct stents were inserted for drainage.The durations of complete disappearance of abdominal pain,decrease of serum and urine amylase values to normal,white blood cell recovery time,liver function recovery time,cost of hospitalization and duration of hospitalization were significantly shorter in the early ERCP group than the control group.The mortalities in the ERCP and the control groups were 8.0％ and 22.2％,respectively.Conclusions Early endoscopic management for patients with acute biliary pancreatitis was not only safe and efficacious,but the management helped to identify the underlying causes of pancreatitis and reduced the duration of hospital stay and expenses.

Objective To investigate galectin3 on proliferation and migration of esophageal cancer Eca109 cells.Methods A lentiviral vector for over-expression of RNA targeting galectin3 was designed to transfect Eca109 cancer cells following plasmid-mediated transfection manual (Eca109/Gal3 cells).Inverted fluorescence microscope was used to observe the expression of EGFP.The proliferation of Eca109 cells was measured by cell counting Kit-8 assay.Eca109 cells apoptosis was determined by Annexin-V/7-AAD doublestaining.The migration capacity of Eca109 cells was determined in transwell assays.Western blot analysis was used to measure the expression of galectin3 protein.Results Galectin-3 expression was detected in Eca109 cells,with the Galectin3 expression in Eca109/Gal3 cells much more than non-transfected cells (t =14.33,P ＜ 0.05 ; t =10.28,P =0.037).Compared with non-transfected Eca109 cells,proliferation increased significantly in Eca109/Gal3 cells (t =-17.277,P ＜ 0.05 ; t =-13.4,P ＜ 0.05).Galectin3 evidently decreased in Eca109 cell apoptosis (t =3.053,P ＜ 0.05 ; t =5.446,P ＜ 0.05).Transwell migration assay showed that a greater number of Eca109/Gal3 cells crossed the artificial basement membrane compared with non-transfected Eca109 cells and negative control Eca109 cells (t =3.465,P ＜ 0.05; t =3.252,P ＜ 0.05).Conclusion Galectin3 expression is detected in transfected esophageal cancer Eca109 cells,whose overexpression can result in enhanced proliferation,migration,invasion as well as reduced apoptosis.These data indicate that in-depth research of galectin-3 may prove to be a potential molecular target for the treatment of esophageal cancer.

Objective To evaluate the therapeutic efficacy of intratympanic dexamethasone injections combined with prednisone in patients with idiopathic sudden sensorineural hearing loss. Methods A total of 71 patients diagnosed with sudden hearing loss were treated with intratympanic dexamethasone injections plus prednisone (B group) or prednisone alone (A group). Hearing was evaluated by pure tone audiogram performed before initial treatment and at 4 weeks following the final treatment. Results The total recovery rate after the treatment was 81.8% in the B group and 55.3% in the A group. The diflference between two groups was statistically significant (P < 0.05). Conclusion The present study suggests that sudden sensorineural hearing loss patients treated with intratympanic dexamethasone combined with prednisone have a higher likelihood of hearing recovery than those treated with prednisone alone.