Objective To investigate the feasibility of transfecting gene into HepG2 by therapeutic ultrasound combined with microbubble,and to explore the optimal ultrasonic parameters for high transfection efficiency.Methods HepG2 cells were cross-interval planted in 24-well plates.EGFP plasmid DNA and microbubbles were added to the cultured HepG2 and three parameters of the therapeutic ultrasound were optimized.Firstly,set ultrasonic intensity gradient (group A,from A1 to A5),which were 0.4 W/cm,0.8 W/cm2,1.2 W/cm2,1.6 W/cm2 and 2.0 W/cm2 respectively.Secondly,set the duty cycle gradient group (group B,from B1 to B3),which were 10％,20％ and 50％ respectively.Lastly,set the exposure time group(group C,from C1 to C3),which were 30 s,90 s and 180 s.After 24 hours,the expression of GFP was observed by fluorescence microscopy,the transfection rates was detected by flow cytometry,the cell death were assessed by trypan blue exclusion.Results The plasmid transfection efficiency was varied under different ultrasonic parameters.Within a certain range,the increasing of parameters can improve the transfection efficiency,but when the parameters were too large (eg.the intensity＞1.6 W/cm2 or 50％ duty cycle),the actual transfection efficiency decreased due to the increased cell death.When the ultrasonic intensity was 1.2 W/cm2,the transfection efficiency and mortality were better than the other subgroups of group A.When the duty cycle was 20％,the transfection efficiency and mortality rate were better than the other subgroups of group B.Continue to increase the exposure time over 90 s was not statistically significant (P ＞0.05) for transfection efficiency and cell mortality.Conclusions The ultrasound-targeted microbubble destruction is an efficient method for gene delivery in vitro.The transfection efficiency vary greatly under different parameters,so optimization parameters is conducive to the promotion of gene transfection.

Objective To investigate the effects of transplantated tumor capillary permeability under microbubble enhanced diagnostic ultrasound exposure.Methods Fourty eight rats were randomly divided into four groups after Walker 256 carcinomas were subcutaneousely implanted,named the control group (A),microbubbles group (B),ultrasound group (C),microbubbles and ultrasound group (D).Using Evans blue(EB) as the indicator for tumor capillary permeability,the microbubbles were injected through the vein with diagnostic ultrasound frequency as 1.75 MHz,mechanical index as 1.4.The spillover of EB was estimated under confocal laser microscope,meanwhile the contents of EB in the tumor tissues were calculated according to the standard curve and spectrophotometry.Results EB spillover was observed around the capillaries in the group D,while EB spillover was only observable on capillary in group A,B and C.The EB content was also significantly higher than those in group A,B and C.Conclusions Exposure to both diagnostic ultrasound and microbubbles results in increased capillary permeability of rat tumor tissues,which might become a potential therapeutic method on clinical chemotherapy.

Objective To evaluate the funding and output of pancreatitis related projects supported by National Natural Science Foundation of China (NSFC),and provide suggestions for the funding strategy in the field of pancreatitis research.Methods Database of funded projects and completion reports of NSFC between 1990 and 2012 were retrieved.The number of grants,amount of grants,types,contents,and paper output of funded projects were respectively analyzed.Results From 2000 to 2012,114 projects involving pancreatitis were funded by NSFC,including 71 general programs (62.3％),8 regional programs,34 youth programs (29.8％),1 key program.The total fund during 2011-2012 reached 25.44 million RMB for pancreatitis research,which was 2.76 times higher than that of 2000-2010.According to PubMed search,93 SCI papers funded by NSFC were published during 2011-2012,which was 2.07 times more than that of 2000-2010.The researches mostly focused on molecular mechanism and therapeutic effect of acute pancreatitis,chronic pancreatitis and severe acute pancreatitis,which accounted for 61.4％ of all projects.Conclusions The number of grants,amount of grants,and NSFC funded SCI paper output of pancreatitis research have grown steadily,and it indicates that long-term funding of NSFC has played a positive role in promoting the development of clinical and basic research in the field of pancreatitis.

Objective To compare bolus infusion and replenishment using real low mechanical index contrast enhanced ultrasound in assessing the change of renal cortical perfusion.Methods Using dopamine (i.v.) at the dose of 0.5,2.0,5.0μg · kg- 1 · min- 1 to change renal blood perfusion of 20 rabbits,then during bolus or contant injection of SonoVue,at coded pulse inversion mode,real-time contrast ultrasound was performed,the latter method needed destroying microbubble at a high MI when amplitude reach a steady state,then recording the replenishment,peak intensity(A) and time to peak (PPT) were obtained through raw time-intensity curve,and slope rate of TIC(k) was acquired by curve fitting,standard effective renal plasma flow(ERPF) was measured through 4-aminohippuric acid clearancerate method,meanwhile correlations between ERPF and parameters were analyzed,as well as the paired samples t test for each parameter before and after dopamine administration.Results The ascending branchs of raw TIC of bolus infusion increased sharply and were approximately straight,then descended gradually,while that of replenishment looked like two straightlines with different slopes,then stayed horizontal Both the value of A of two methods were positively correlated with ERPF ( r b =0.85,r re =0.66),and were different at the same ERPF,meanwhile the value of TTP were negatively correlated with ERPF( r b =-0.92,r re =- 0.76),and there were no statistically difference between the two methods.k from Gamma fitting was far from correct,while k from exponential fitting was apparently correlated with ERPF ( r re =0.77 ).Conclusions Both bolus injection and constant injection-replenishment method can assess renal cortical blood perfusion,TIC parameters A and TTP represent regional blood volume fraction and microbubble velocity respectively.Bolus-infusion with real low mechanical index is more precise and available.Comparing with k,TTP is more appropriate to reflect perfusion velocity.

Objective To study the current management strategy for colorectal injuries. Methods The clinical data of 86 consecutively admitted patients with penetrating colorectal injuries were retrospectively reviewed. Results Most injuries were of closed type. Iatrogenic colon injury accounted for 14%. There were 61 patients (71%) associated with other abdominal organ injuries. Operations were performed in all patients. Seventy six patients were treated with primary repair or resection and anastomosis, and 10 with diverting colostomy. In recent 10 years mortality rate dwindled from 10% to 4%(2/46). Mortality was most often caused by hemorrhagic shock, associated organ injuries or severe secondary infection. Conclusions Most colorectal traumas are blunt closed type in China. Primary repair or resection and anastomosis at the time of initial exploration is most often used for colorectal injuries. Indications for diverting colostomy are severe shock, heavy intra abdominal contamination, poor condition, and rectal injury. Fecal peritonitis should be effectively prevented and treated.

A management window model for patients with atrial fibrillation (AF) was established in Tilanqiao Community Health Service Center cooperated with tertiary hospitals.Patients were screened and treated in the community health service center,and a comprehensive management plan was conducted and patients were followed-up.A total of 105 patients with atrial fibrillation (97.2％) were effectively managed with an average follow-up of 10.6 months.The CHADS2 score ≥ 2 in 78 cases,including 11 cases with administration of warfarin (14.1％) and 26 cases with paroxysmal AF without transition to persistent atrial fibrillation.Compared to the data before management,the rate of taking aspirin and warfarin in managed patients was increased (all P ＜ 0.05),the international normalized ratio (INR) of patients receiving warfarin was improved (P ＜ 0.05) ; the ratio of receiving comprehensive treatment program,standardized treatment and health education were significantly increased (P ＜ 0.05) ; the awareness of the disease,treatment compliance and satisfaction of patients were improved significantly (all P ＜ 0.05).The results suggest that the management widow model is feasible and effective for management of patients with atrial fibrillation in community health service centers.

Objective To evaluate atrial septal defect (ASD) occlusion employing a small right anterior thoracotomy approach. Methods A total of 21 patients with ASD underwent general anesthesia and 2 -3 cm incision was made in the fourth right intercostal space. Utilizing transesophageal or transthoracic echocardiography, the occluder was released using a monotube unit. Results All patients were occluded successfully. No patient required open surgery utilizing extracorporeal circulation. There were no major complications and no evidence of residual atrial shunt. Conclusion ASD occlusion via a minimal surgical incision is safe, less invasive, and has excellent outcomes.

BACKGROUND: Differentiation of bone marrow stromal cells (BMSCs) into neurons requires two processes: orientation and differentiation. Orientation and differentiation are results from different gene expression in cells with the same gene bank. Gene expression requires a certain condition. Changes in extracellular matrix can induce changes in cell morphology and gene expression manner. OBJECTIVE: To explore the possibility of BMSC differentiation into neuron-like cells under tissue extract from rat damaged brain. METHODS: The fifth passage of green fluorescent protein (GFP)-transfected BMSCs was induced to differentiate in brain tissue extract from ischemia/reperfusion rats or normal rats. A blank control was set. Cell morphology change was observed under the phase contrast microscope, and then evaluated using immunohistochemical staining. RESULTS AND CONCLUSION: Primarily cultured BMSCs were purified and amplified, and then showed even spindle shape. The third passage of BMSCs was positive for CD44 and CD106, but negative for CD34. Under the fluorescence microscope, BMSCs showed fluorescence expression, but the strength was weak 24 hours following GFP transfection. Numerous cells presented significant green fluorescence 48 hours later. Following adding brain tissue extract from ischemia/reperfusion rats. Induced cells presented neuron-like feature, but neuron specific enolase specific antibody presented positive expression. Compared with the blank control group, the differentiation rate of BMSCs was significantly increased in the ischemia/reperfusion group and normal group (P < 0.05). The increased range was significantly greater in the ischemia/reperfusion group than the normal group (P < 0.05). These results indicated that brain tissue extract from ischemia/reperfusion rats can successfully induce the differentiation of BMSCs into neuron-like cells.

Objective:To investigate whether His-tag to change the immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus.Methods:The G1 and F/M2 gene fragments were amplified by PCR method and then ligated into the expressing vector pET-His or pET-DsbA-His.Each recombinant plasmid was transferred into E.coli BL21(DE3) and the expression was induced by IPTG.The expressed His-G1F/M2 or DsbA-His-G1F/M2 was purified by affinity chromatography.The latter was digested with thrombase and G1F/M2 was purified by affinity chromatography.His-G1F/M2 or G1F/M2 was used to immunize BALB/c mice.Anti-RSV antibody was measured by ELISA and RSV-specific CTL responses by MTT.Results:No significant difference was observed between the level of anti-RSV antibody or RSV-specific CTL response induced by G1F/M2 and that by His-G1F/M2.Conclusion:His-tag does not change the immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus.

Objective To analyze the status of mycoplasma infection and drug resistance in the local area,and provide the ba-sis for clinical rational drug use.Methods The specimens obtained from 1 9 530 patients with urogenital tract infection were detec-ted by adopting mycoplasma culture,identification and drug sensitivity integration kit.Mycoplasma infection and drug susceptibility were analyzed.Results In the total of 1 9 530 suspected patients specimens,1 1 1 78 cases were positive with a positive rate 57.24%.The positive rate of ureaplasma urealyticum (Uu)and mycoplasma hominis (MH)was 44.63% and 0.44% respectively and the positive rate of Uu and Mh mixed infection was 12.1 7%.The positive rate of female was higher than that of male and the difference was statistically significant(P <0.05).The positive rate of mycoplasma in 2008-2014 was on the rise;The sensitive rate of mycoplasma to josamycin,doxycycline,minocycline element,clarithromycin was 88.57%,84.32%,76.09% and 71.53% respec-tively,mycoplasma was highly drug resistance to quinolone antibiotics;mixed infection resistance was higher than that of single in-fection;The number of drug resistance of Uu,MH and Uu+MH to 12 kinds of antibiotics increase.Conclusion Mycoplasma infec-tion in urogenital tract is mainly caused by Uu and Mh infection is in mixed infection way;josamycin,doxycycline is the first choice for treatment of mycoplasma in this region.Rational drug choise can be based on the drug susceptibility test.Multiple drug resist-ance of mycoplasma is serious and should be paid attention to.