Objective Clearing HIV provirus is the key to cure AIDS .The study was to construct the Tre enzyme eukaryotic expression vector and identify its function in specific recognition of loxLTR sequence in HIV provirus . Methods Tre gene was in-serted into eukaryotic expression vector pcDNA 3.1 gene recombination manipulation by genetic recombination techniques including gene synthesis , PCR, restriction enzyme digestion and ligation .EGFPpA-LoxLTR sequence was inserted into pmCherry-N1 vector and was tested by restriction enzyme digestion , PCR and sequencing .Constructed vectors were electroporated into HeLa cells , then using fluorescence microscopy to observe fluorescence intensity changes . Results PCR, restriction enzyme digestion , electrophoresis and sequencing confirmed that Tre enzyme eukaryotic expression vector had been constructed successfully , and it could specifically recog-nize and cut loxLTR sequence after being transfected into Hela cells . Conclusion Constructed Tre enzyme eukaryotic expression vector can be expressed in Hela cells and specifically recognize loxLTR sequence , which has prepared the experimental ground for fur-ther studies of clearing HIV provirus .

Objective To investigate standardization of technique of retroperitoned laparoscopic ureterolithotomy.Methods Retroperitoneal laparoscopic ureterolithotomy was performed on 85 cases of ureterolithiasis(the stone size ranging from 0.8~2.5cm,40 cases in the left,45 cases in the right;and 25 had undergone unsuccessful ESWL).Results 84 cases succeeded,1 case opened becase of play D-J tube difficultly,the mean operation time was 75 minutes(48~220min),and the estimated blood loss 20~100ml,the drain tube was removed 5~12 days(mean 7 days),1 case experienced urinary leakage,1 case infective in the retroperitoneal.Conclusion Retroperitoneal laparoscopic ureterolithotomy was a safe operation,it can take the place of open surgery.

ObjectiveTo analyze the technique and clinical effect of percutaneous nephrolithotomy combined with endoscopic balloon dilation in the treatment of upper ureterostenosis with recurrent renal calculi. MethodsFrom June 2008 to June 2011,18 ureteral stenosis patients with the history of ureteral open surgery,postoperative residual or recurrent kidney stones were treated.There were 8 males and 10 females with the age of 27 -48 years.Fourteen cases were with hydronephrosis of 2 -4 cm,3 cases were with hydronephrosis of 5 -6 cm and 1 case was with hydronephrosis ＞6 cm.Subsequent stone size ＜ 1 cm was found in 15 cases,1 -3 cm in 3 cases,＞3 cm in 1 case.All patients were treated with percutaneous nephrolithotomy ultrasonic lithotripsy combined with balloon dilatation.The stone clearance rate,hydronephrosis changes,complications and IVP situation before and after surgery were analyzed.ResultsAll the 18 cases were completed surgery successfully.There was 1 (6％) case with renal hemorrhage 3 days after the surgery and controlled with DSA hemostasis.There was 1 case accepted adjusting double-J tube by ureteroscopy.Sixteen (89％) patient's stones were completed removed.One case with residual calyceal stones size ＜5 mm was not further treated.There was 1 case treated with nephrectomy because of renal stone with infection.The patients were followed up for 6 to 36 months.Fourteen cases with hydmnephrosis improved significantly; 3 cases with no significant changes but improved following balloon dilation.All patients achieved significant improvement in imaging study comparing of preoperative and postoperative data.ConclusionThe use of percutaneous nephrolithotomy combined with endoscopic balloon dilation is a safe and efffective treatment option in the treatment of kidney stones with ureteral stenosis.

Objective To investigate the feasibility of urethral reconstruction with colonic mucosa graft in the treatment of complex long urethral stricture.Methods The clinical data of three cases with complex long urethral stricture were reported and analyzed.Patient ages were 71,64 and 48 yrs and the course of disease was three months,six months and six yrs,respectively.The length of urethral stricture was 13,18 and 12 cm.Removing the narrow urethral segment and intercepting the length from 12 to 18 cm sigmoid colon and stripping colonic mucosa were performed.Urethral reconstruction was done with a free graft of colonic mucosa.Follow-up included urethrography,uroflowmetry,and urethroscopy.Results The urethral reconstructions were completed successfully.The urinary peak flows of the patients were 16.7 ml/s,19.6 ml/s and 26.4 ml/s at six weeks post operation.Urethrography revealed the graft urethral lumens were bulky three months after the operation.In urethroscopy,the colonic mucosa was found to be of good color and the anastomotic site healed well.Patients were followed-up 28,16,and three months,respectively,and were all voiding well.Conclusions Colonic mucosa graft urethroplasty is a feasible procedure for the treatment of complex long urethral stricture.

More and more DNA sequences have been obtained since the start-up of human genome project. Powerful system is badly needed for data mining on these DNA sequences. Based on a personal computer and Linux operating system, the Phred/Phrap/Consed software and Blast software were used to construct a platform for batch analysis of the sequences, including identifying raw DNA sequence from chromatogram file, vector sequence removing, contig analysis (sequence assembly), repeat sequence identifying and sequence similarity analysis. Result demonstrated that this robust platform could accelerate data analysis for large-scale DNA sequencing.

Objective To observe the characteristic changes in cardiac function and to raise diagnostic accuracy and sensibility.Methods 39 silent myocardial ischemia and 58 angina and 46 age-matched healthy subjects and 62 obsolete myocardial infarction underwent myocardial tomography imaging and gated ventricular imaging.Covariance analysis was used to compare difference between SPECT imaging and parameters of function.The age was tested as covariate factor,then SNK-Q test and multiple comparison were carried on.Results The silent myocardial ischemia and angina showed reversible radioactive defect or fixed defect.Most of myocardial SMI showed single peak exempt TPFR.EF,PFR and PER showed remarkably statistical difference (P<0.01).The multiple comparison of PFR and PER was increased according by OMI,AN,SMI and negative control group (P<0.05).Conclusions The myocardial damage and PFR,PER in SMI is lower than that in AN group,which have no remarkably influence on the parameters of diastolic and retractile function in left ventricular.

Objective To retrospectively analyze the surgical methods and efficacy in 70 cases of type A aortic dissection patients over 65 years old.Methods From January 2005 to May 2012,70 type A aortic dissection patients over 65 years old received surgical treatment.Among them,there were 47 males aged 65 to 78 years old with mean 71,23 females,aged 65 to 72 years old with mean 68.55 cases were acute onset,while 15 cases were chronically onset.Different surgical methods were selected depend on patients' situations.We followed up all patients after discharged from hospital to continue to observe their health situation and evaluate the therapeutic effects.Results After surgery,eight patients died in the hospital,62 patients were recovered and discharged from the hospital.The mortality rate is 11.4％.During the follow up period from 3 to 72 months,there were no dead,aneurysm rupture and others severe complications.9 cases received endovascular graft exclusion within 6 months after discharged from hospital.The survival patients were satisfactory healed with their daily living activity resumed.Conclusion For over 65 years old patients with aortic dissection,the accurate and rapid selection of surgical method could improve the survival rate and the quality of life with a lower occurrence rate of complications.

OBJECTIVETo compare the efficacy of telbivudine monotherapy and telbivudine combination therapy with adefovir in patients with nucleos(t)ide-naive chronic hepatitis B, high-level hepatitis B virus (HBV) load and hepatitis B e antigen (HBeAg)-positivity, and to explore the relationship between treatment regimen adherence and treatment outcomes.

METODSA retrospective study was performed with 123 HBeAg-positive, high-level viral load (HBV DNA≥10(7) copies/ml), nucleos(t)ide-naive chronic hepatitis B patients. Fifty-three of the patients received combination therapy with telbivudine and adefovir dipivoxil,while 70 patients received the telbivudine monotherapy. All patients were tested for rates of conversion to HBV DNA-negative status,alanine aminotransferase (ALT) normalization, HBeAg seroconversion, drug resistance, and side effects at treatment weeks 12, 24, and 48. Treatment regimen adherence was assessed through self-reporting, and interviews were used to explore the relationships to treatment outcomes. The chisquare test, t test and Fisher's exact test were used for statistical analyses.

RESULTSThe rates of HBV DNA negative conversion in the combination group at treatment weeks 12, 24 and 48 were 62.3% (33/53), 88.7% (47/53) and 94.3% (50/53) and were significantly different from those in the monotherapy group at weeks 12 and 24.The rates of ALT normalization were significantly different between the two groups at treatment week 12 (94.3% vs. 77.1%). The rate of HBeAg seroconversion in the combination group at treatment week 48 was 39.6%, and significantly different than that of the monotherapy group. The rates of drug-resistance in the combination and monotherapy groups at treatment week 48 were 3.8% and 11.4%, and the proportion of non-adherence to the treatment regimen was 53.3%, which significantly affected treatment outcome. No side effects occurred in either treatment group.

CONCLUSIONTelbivudine combination treatment with adefovir was more effective than telbivudine monotherapy and elicited a low drug resistance rate in nucleos(t)idenaive chronic hepatitis B patients with high-level HBV load and HBeAg-positivity. Adherence to the therapy regimen was a key factor influencing treatment outcomes.

Adenine ; analogs & derivatives ; Alanine Transaminase ; Drug Therapy, Combination ; Hepatitis B e Antigens ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Organophosphonates ; Retrospective Studies ; Thymidine ; analogs & derivatives ; Treatment Outcome ; Viral Load

Objective To explore the effect of sucralfate on cytokines in rats with paraquat (PQ) poisoning. Methods Seventy-two healthy male Sprague-Dawley (SD) rats were randomly divided into PQ model group, sodium bicarbonate intervention group (SB group) and sucralfate suspension gel group (LTL group), with 24 rats in each group. The rat model of PQ poisoning was reproduced by one-time intragastric administration of PQ solution 25 mg/kg. The rats in SB group and LTL group were intragastricly administrated with 5 mL·kg-1·d-1of 100 g/L sodium bicarbonate or 200 g/L sucralfate at 2 hours after exposing to PQ, and the rats in PQ model group were given the same amount of sterile saline. The abdominal aortic blood of rats was collected at 1, 3, 6, and 10 days after PQ poisoning, and the levels of serum tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) were determined by enzyme-linked immunosorbent assay (ELISA). The left lung tissue was harvested, and lung wet/dry weight (W/D) ratio was assessed. Results With prolonged exposure, lung W/D ratios in all the groups were increased gradually, reached the peak at 10 days, but in the SB group and LTL group, the amplitude of increase was obviously reduced, the ratios were significantly decreased at 6 days and 10 days as compared with those in PQ model group (SB group vs. PQ model group: 4.99±0.79 vs. 6.98±0.86 at 6 days, 5.61±0.36 vs. 7.36±0.95 at 10 days; LTL group vs. PQ model group: 4.61±0.24 vs. 6.98±0.86 at 6 days, 4.24±0.20 vs. 7.36±0.95 at 10 days, all P < 0.05), but there was no significant difference between SB group and LTL group (all P > 0.05). After PQ poisoning, the levels of TNF-α, IL-10 and TGF-β1 were elevated, and reached the peak at 3 days and then decreased gradually. Compared with the PQ model group, serum TNF-α, IL-10 and TGF-β1 levels in SB group and LTL group were decreased significantly [SB group vs. PQ model group: 3-day TNF-α (ng/L) was 147.6±12.3 vs. 168.2±11.3, 3-day IL-10 (ng/L) was 65.4±3.2 vs. 115.1±9.2, 3-day TGF-β1 (ng/L) was 356.3±50.3 vs. 415.6±68.3; LTL group vs. PQ model group: 3-day TNF-α (ng/L) was 82.2±7.4 vs. 168.2±11.3, 3-day IL-10 (ng/L) was 44.4±5.2 vs.115.1±9.2, 3-day TGF-β1 (ng/L) was 296.3±40.2 vs. 415.6±68.3, all P < 0.05], especially in LTL group (all P < 0.05). Conclusion Early gastrointestinal lavage with sucralfate could effectively reduce the inflammatory exudation in lung tissue after PQ poisoning, and inhibit the cytokine secretion.