Objective To prepare a targeted ultrasound micro bubble,which carried the HSV-TK gene,and investigate the in vitro target searching ability of the micro bubbles and inhibitory effect on the proliferation of HepG2 cells.Methods Ultrasonic micro bubbles were prepared by mechanical vibration method,construction of targeted HSV-TK ultrasound micro bubbles by biotin affinity bridge construction.To detect the general characteristics of ultrasound micro bubbles,and to test its effect on the proliferation of HepG2 cells in vitro.Results HSV-TK targeted ultrasound microbubbles more gathered on the surface of HepG2 cells,through detection of PCNA and MTT,it was found that the proliferation of gene targeting microbubble group was obviously decreased,cell apoptosis increased significantly,Cells invade experiments showed that the number of cells in genetic microbubble group (22.18 ± 2.01) decreased significantly compared with the control group and the nontargeted group,can effectively inhibit the proliferation and invasive ability of HepG2 cells.Conclusion Targeted ultrasound microbubble carrying target gene have better inhibitory effect on HepG2 cells in vitro.

Background and purpose:Tumor metastasis is a main reason of breast cancer patients’ death. This study aimed to discuss whether or how the transcription factor ETS2 regulate CXCR4 transcription and the molecular mechanism of ETS2 modulating transcriptional activity of CXCR4 gene in human breast cancer cells. Methods:In MCF-7 breast cancer cell lines and MDA-MB-231 breast cancer cell lines, through transient transfection, as well as RNAi technology, the expression of ETS2 was overexpressed or inhibited was detected. RT-PCR and ELISA was used respectively to detect CXCR4 mRNA expression and protein level. Luciferase reporter gene assay was applied to detect CXCR4 promoter activity, and ChIP for detecting the amount of ETS2 protein binding to CXCR4 promoter. Two binding sites of CXCR4 promoter were mutated to detect the impact on the activity of CXCR4 promoter by gene mutations. Results:After transfected with ETS2 expression vector in MCF-7 and MDA-MB-231 breast cancer cell lines, the mRNA expression and protein level of CXCR4 were elevated. The result of luciferase reporter gene assay indicated that overexpression of ETS2 activated CXCR4 promoter. ChIP assay demonstrated that the amount of ETS2 protein binding to CXCR4 promoter increased after ETS2 transfection. This result indicated that ETS2 may activate CXCR4 promoter through directly binding with CXCR4 promoter. Inhibition of ETS2 expression using RNAi could significantly attenuate CXCR4 promoter activity and reduce expression of CXCR4. Two ETS binding sites of CXCR4 promoter were mutated and the result of luciferase reporter gene assay proved that, an arbitrary point mutations attenuated CXCR4 promoter activity, while mutation of both binding sites further attenuated CXCR4 activity. Conclusion:In MCF-7 and MDA-MB-231 breast cancer cell lines, overexpression of ETS2 could activate CXCR4 promoter and the transcription of CXCR4 through directly binding to two ETS2 binding sites (-540 to-535 and-240 to-235) of CXCR4 promoter.

Objective To explore the effect of neural stem cells (NSCs) transplantation on neurogenesis in an alzheimer' disease mouse model and its related mechanism.Methods A total of 16 12-month old aPP/PS1 double transgenic aD mice were randomly divided into two groups:NSC group (receiving NSCs transplantation in the bilateral hippocampi,n=8) and PBS group as the negative control group (receiving an equal quantity of 0.01M/L phosphate buffer saline,n=8).another 8 wild type mice without any treatment were selected as the positive control group (Wt group).at five weeks after transplantation,the expressions of doublecortin (DCX) and nestin in the hippocampal dentate gyrus (DG) and subgranular zone (SGZ) were analyzed by immunofluorescence staining,the number of positive cells in these regions were counted by confocal microscopy,and Morris water maze (MWM) test was used to assess cognitive function in all mice.Results NSC group showed the enhanced spatial learning and memory ability in Morris water maze (MWM) as compared to PBS group,but it was still lower than that in Wt mice (both P＜0.05).Immunofluorescence staining showed NSCs induced the increased positive cells expressing DCX and nestin in the NSC group as compared with PBS group (F＝85.400 and 55.687,both P＜0.001).However,the number of DCX positive cells was still lower in NSC group than in Wt group (P＜0.05).There was no significant difference in the number of Nestin-positive cells between the NSC group and the Wt group (P＞ 0.05).Conclusions NSCs transplantation can promote endogenous neurogenesis via the increased expression of DCX-and Nestin positive cells in the hippocampal DG/SGZ in aD mice,which improves the cognitive abilities in aD mice in some extent.

OBJECTIVE:To investigate the effects and safety of Compound xuelian burning wound ointment in the treatment ofⅡdegree deep burn. METHODS:Using consubstantial control method,2 burn wounds(Ⅱdegree deep burn,about 10 cm×10 cm) selected from symmetric or adjacent part of extremities in 80 patients withⅡdegree deep burn were divided into treatment area and control area. Treatment area was given Compound xuelian burning wound ointment,and control area was given Sulfadiazine silver cream. Healing time,degree of pain due to dressing change,the amount of infiltration liquid,wound infection rate,the incidence of scar formation and the incidence of ADR were observed. RESULTS:The healing time of wound in treatment area was signifi-cantly shorter than in control group [(18.7±3.6)d vs.(23.8±3.1)d],with statistical significance(P<0.05). During treatment,the proportion of patients suffering from acute pain and severe pain due to dressing change in treatment area,the amount of infiltration liquid and the incidence of scar formation were significantly lower or less than in control area;the proportion of patients suffering from mild pain due to dressing change in treatment area was significantly higher than in control area,with statistical significance (P<0.05). There was no statistical significance in the proportion of patients suffering from moderate pain and indolence due to dressing change,the incidence of wound infection between treatment area and control area(P>0.05). No obvious ADR was found in 80 patients. CONCLUSIONS:Compound xuelian burning wound ointment in the treatment of Ⅱdegree deep burn can relieve the degree of pain due to dressing change,control the amount of infiltration liquid,promote wound healing and reduce the occurrence of scar with good safety.

The new intein-mediated PHB purify protein system is a high expression, automatic cutting, for purification, low-cost protein purification system,it is conducive to large-scale protein purification.Choose human antibacterial peptide LL-37 as the purification objects,which is poison to prokaryotic cell.We construct intein-mediated PHB purified human antimicrobial peptide LL-37 system through genetic engineering technology and use this system to purify LL-37. The results show that this system can highly express LL-37 fusion protein and purifiy the product as same size with expectations.

Objective To investigate a new single nucleotid polymorphism (SNP) intron5(+4668C/T) in SLC22A12 in primary gout patients and the association between clinical characteristics and genotypes. Methods One hundred and one primary gout patients and 186 healthy subjects were recruited into this study. Blood pressure, body mass index (BMI) was recorded. Serum uric acid, glucose, lipid and creatinine were detected. DNA was extracted from peripheral blood to amplify the fragment located in intron 5. The genotypes of SLC22A12 can be detected with high-resolution melting (HRM) assay, followed by sequencing analysis. Chi-square test was used for statistical analysis. Results ① A new SNP in intron 5 of SLC22A12 was identi-fied successfully by HRM, which was defined as intron 5 (+4668C/T). CC, CT and TT genotypes were unam-biguously distinguished with HRM technology, which was fully concordant with sequencing. ②The genotypes of CC, CT and TT in male and female groups were 28.1%, 33.7%, 38.2% and 20.0%, 47.1%, 32.9%, respectively.③ However, no significant differences of genotype distribution were found concerning BMI, blood pressure, creatinine, total cholesterol and triglyceride in both male group and female group. But the serum uric acid levels in the CC genotype were significantly higher than those with the CT+TT genotypes. ④ The genotype frequencies of CC and CT+TT in high uric acid group were remarkably different from those in low uric acid group (21.2%, 78.8%,; 35.0%, 65.0%; P<0.05). Conclusion A new SNP has been successfully discovered with HRM technology with simplicity, rapidity and accuracy. T allele of intron 5 (+4668C/T) may be a genetic protective factor for hyperuricemia among Chinese population.

Objective To evaluate the efficacy of preoperative biliary drainage (PBD) in the pancreaticoduodenectomy for malignant obstructive jaundice.Methods Database including PubMed,EMBASE,Cochrane Central Register of Controlled Trials,Academic Degree Dissertation Database and Conference Database were searched with malignant obstructive jaundice,pancreaticoduodenectomy,preoperative biliary drainage,comparative study.Literatures about the randomized controlled trials of PBD (PBD group) and efficacy of early surgery (ES group) in the pancreaticoduodenectomy were retrieved from January 2001 to December 2013,and then a Meta analysis was carried out based on the data.The count data were analyzed using the odds ratio (OR),relative risk (RR) and 95％ confidence interval (95％ CI),and the measurement data were analyzed using mean difference (MD) and 95％ CI.The heterogeneity of the data was analyzed using the I2 test.Data were integrated by fixed or random effect model.Results Twelve literatures including 1 982 patients were selected.There were 1 029 patients in the PBD group and 953 in the ES group.The results of Meta analysis showed that the operation time,volume of blood loss and rate of postoperative wound infection in the PBD group were significantly different from those in the ES group (MD =10.50,107.92,95％ CI:6.34-14.66,16.43-199.42;RR =1.62,95％CI:1.19-2.21,P ＜0.05).There were no significant differences in the postoperative mortality,incidence of pancreatic fistula,incidence of bile leakage,incidence of delayed gastric emptying and duration of hospital stay between the 2 groups (RR=0.69,95％CI:0.52-0.92;OR =0.68,1.35,95％CI:0.38-1.21,0.93-1.95;MD =0.69,95％CI:-0.67-2.05;RR =0.00,95％ CI:-0.02-0.01,P ＞0.05).Conclusion PBD in the pancreaticoduodenectomy for malignant obstructive jaundice cannot reduce postoperative mortality and incidence of complications in patients,and should not be used as the conventional management in the perioperative period.

Objective To evaluate the short-term prediction of high-sensitivity cardiac troponin T (hs-cTnT) and other cardiovascular risk biomarkers in patients undergoing maintenance hemodialysis (MHD).Methods We conducted a cohort survey in 296 consecutive MHD patients whose clinical data were retrospectively analyzed.Before MHD,hs-cTnT and other relative cardiovascular biomarkers were detected.The end point (all-cause death) and time of occurring were recorded in the next 13 months.The differences between survival and all-cause death were analyzed by t-test,Mann-Whitney test and x2 test.The best two percentile cutoff point was calculated by X-tile and the survival rate was calculated by Kaplan-Meier Logistic regression analysis was applied to analyze the odd ratio between high risk and non-high risk hs-cTnT group.Non-high risk group was divided into intermediate risk and low risk group based on the 99th percentile of hs-cTnT in healthy population,to further evaluate its short-term prediction value for MHD patients.The short-term significance of hs-cTnT was proved to be independently associated with all-cause death by Logistic regression analysis.Results The mean value of serum hs-cTnT in survival group was 0.05 (0.03～0.07) ng/mL,while in the death group it was 0.07 (0.04～0.14) ng/mL,which had statistical significance (P =0.027).The best two percentile cutoff of hs-cTnT in MHD patients was 0.1 ng/mL.The survival rate in high risk group (hs-cTnT＞0.1 ng/mL) is lower than it in non-high risk group (hs-cTnT≤0.1 ng/mL) (76.67％ vs.96.62％,P ＜0.05).The odd ratios for high risk group and non-high risk group was 7.288 (P＜ 0.001).Moreover,further grouping the non-high risk group by hs-cTnT =0.014 ng/mL,intermediate risk group (hs-cTnT＞0.014 ng/mL) group has lower survival rate than low risk group (hs-cTnT≤0.014ng/mL),while there wasn't any death case occurred in the low risk group.Conclusions Hs-cTnT is an independent risk factor to all-cause death.Thus hs-cTnT can be a strong indicator of short-term prediction and prognostic evaluation.

Objective:To evaluate the clinical effects of invisible bracketless appliance on the upper molar distalization.Methods:15 patients with class Ⅱ malocclusion(at the average age of 25.3 years) were treated by invisible bracketless appliance,the pre and post three-dimensional digital models were superimposed and measured,the effects of molar distalization were analysed.Results:After treatment,the maxillary first molars were distalized by 2.58 mm on both sides.The left and fight second molars were distalized by 2.57 mm and 2.68 mm respectively.Bilateral central incisors were moved mesially by 0.34 mm.There was no significant difference in the horizontal movement of central incisor.But the left and fight first molars were moved buccally by 0.96 mm and 0.97 mm respectively,the left and fight second molars were moved buccally by 1.01 mm and 1.11 mm separately.Bilateral first molars were intruded by 0.26 mm,the left and right second molars were intruded by 0.37 mm and 0.36 mm,respectively.But the central incisors had no significant vertical movement.There was no significant difference in the buccally or palatally rotation of bilateral first and second molars.Conclusion:Invisible bracketless appliance is efficient for distalization of upper molar,but it may result in mild molar intrusion and anterior anchorage loss.

Objective To investigate the clinical value of endoscopy in the treatment of choledocholithiasis. Methods All 227 patients underwent endoscopic treatment. 14 of them with stones less than 1. 0 cm in diameter were treated with endoscopic papillary balloon dilation; 194 patients with stones 1. 0-1. 5 cm in diameter were treated with endoscopic sphincterotomy; and 19 patients with stones up to 1.5 cm in diameter were treated with endoscopic mechanical lithotripsy. Results Choledocholithiasis were not confirmed by choledochography in 34 cases ( 15% , 34/227) , who underwent exploration of common bile duct through EPBD or EST, but no stones were found. In 187 of the 193 choledocholithiasis patients their stones were removed, the overall success rate with complete stones clearance was 96.9% ( 187/193). The incidence of complication was totally 5. 29% ( 12/227) , including acute cholangitis in 3 patients, acute pancreatitis in 8 patients, and bleeding in 1 patient. Conclusions Endoscopic treatment should be the first choice of patient with choledocholithiasis due to its advantages of safety, effectiveness and with less complication. EPBD or EST was not recommended in case of the common bile duct stones were not confirmed by ERCP, in order to avoid the risk of papilla injury.