Objective:To construct eukaryotic expression vector for hepatitis B virus X(HBV X) gene with enhanced green fluorescence protein(EGFP),and confirm its expression in hepatocellular carcinoma(HCC)cell line.Method: HBV X gene was cloned from pcDNA3.1(+)-X by enzyme digestion and inserted to pEGFP-N1.The rector was confirmed by enzyme digestion,PCR assay and sequencing.Then pEGFP-X was transfected to HCC cell line Bel-7402.After transient transfection,expressions of HBV X and EGFP gene were detected by RT-PCR and fluorescence microscope respectively.Results: The results showed that the recombinant plasmid could express HBV X gene efficiently in Bel-7402,which showed green fluorescence.Conclution: The pEGFP-X was constructed and the fused HBV X-EGFP gene was expressed in Bel-7402 successfully.These facilitate the study of the effect of HBV X protein on the development of HCC.

Objective To observe the effect of leflunomide and cyclophosphamide combined with prednisone in the treatment of idiopathic membranous nephropathy .Methods 30 patients with idiopathic membranous nephropa-thy were randomly divided into two groups ,15 cases in each group .A group was treated by leflunomide combined with prednisone , B group was treated by cyclophosphamide combined with prednisone , 6 months a course .The clinical effect,24h urine protein quantity,serum albumin levels,blood fat,renal functions and adverse reactions were com-pared.Results 3,6 months after treatment ,the remission rates in A group were 60.00%,73.33%,those in B group were 53.33%,66.67%(χ2 =0.965,0.896,all P>0.05).After treatment,24h urine protein quantity,serum albu-min levels in the two groups [A group:(1.33 ±1.25)g/24h,(38.24 ±4.84)g/L;B group:(1.42 ±1.37)g/24h, (37.12 ±5.43)g/L] were significantly lower than those before treatment [A group:(7.34 ±2.75)g/24h,(20.31 ± 7.33)g/L;B group:(7.22 ±2.84)g/24h,(20.46 ±7.73)g/L] (A group:t=6.232,5.734,all P<0.05;B group:t=5.934,5.267,all P<0.05).After treatment,total cholesterol,triglyceride in the two groups [A group:(6.74 ± 1.45)mmol/L,(2.43 ±0.75)mmol/L;B group:(6.63 ±1.45)mmol/L,(2.14 ±0.63)mmol/L] were significantly lower than those before treatment [A group:(11.94 ±2.98) mmol/L,(3.56 ±1.35) mmol/L;B group:(11.53 ± 2.84)mmol/L,(3.24 ±1.46)mmol/L] (A group:t=6.246,5.234,all P<0.05;B group:t=5.256,5.672,all P<0.05).There were no significant differences in urea nitrogen ,serum creatinine between before and after treatment in the two groups(all P>0.05).There were no significant differences in above indicators between A group and B group after treatment (all P>0.05).The incidence rate of adverse reactions in A group (13.33%) was significantly lower than that in B group(40.00%)(χ2 =4.246,P<0.05).Conclusion The clinical efficacy of leflunomide is similar to cyclophosphamide in treating idiopathic membranous nephropathy with fewer adverse reactions .

Objective To investigate the therapeutic effect of human umbilical cord blood nucleated cell therapy(HUCBNCT) in patients with chronic severe viral hepatitis,and the effect of HUCBNCT on ductular proliferation and hepatic oval cell proliferation.Methods A total of 90 patients with chronic severe viral hepatitis were divided into two groups randomly,and the two groups were treated with adult plasma transfusion(control group),and HUCBNCT(treatment group),respectively.The therapeutic effect in all patients was evaluated by determination of liver function.Some patients had liver biopsy and CK19,CD34,albumin and AFP of the biopsy tissues were detected.Results Liver necrosis in treatment group was milder than that of control group,and the positive rate of CK19 staining in treatment group was higher than that of control group.Co-expression of CK19 and albumin were observed in some atypical ductular proliferation cells,which were like the oval cells in shape and immunohistochemical characteristics.Conclusion HUCBNCT,which can promote liver ductular and oval cell proliferation,may have promising therapeutic effect on patients with chronic severe viral hepatitis.

OBJECTIVE To explore the relationship between cytokines(IL-1?,mIL-2R,IL-10) and hepatatis B virus(HBV) serum markers in patients with chronic hepatitis B(CHB).METHODS The serum level of IL-1?,mIL-2R and IL-10 in 65 patients with CHB was measured with chemiluminescence immunoassay(CLIA).RESULTS Of the patients with positive and negative HBeAg,the serum IL-1? and IL-10 levels were significantly higher than normal controls(P

Objective To understand the risk factors for poor prognosis of severe adenovirus pneumonia (SAP)in children,and provide guidance for clinical diagnosis and prognosis.Methods Clinical data of 91 hospitalized children who diagnosed with SAP in Chongqing Children’s Hospital of Chongqing Medical University between January 2012 and January 2014 were analyzed retrospectively.Results Of 91 SAP children,23 (25.27%)had poor prognosis. Univariate analysis showed that risk factors for poor prognosis of SAP were age of onset,congenital heart disease and other serious underlying diseases,mechanical ventilation therapy,acute respiratory distress syndrome (ARDS), atelectasis and other serious radiological changes,and emergence of two and more extra-pulmonary complications (all P <0.05).Multivariate regression analysis showed that congenital heart disease and other serious underlying diseases,and emergence of two and more extra-pulmonary complications were independent risk factors for poor prognosis of SAP (all P <0.05).Conclusion Congenital heart disease and other serious underlying diseases,emer-gence of two and more extra-pulmonary complications are independent risk factors for poor prognosis of SAP,active intervention should be conducted during the process of clinical diagnosis and treatment,so as to improve the prognosis.

Objective To evaluate therapeutic effects of radiofrequency ablation for thyroid benign nodules . Methods By using the radiofrequency electrode (18G ×7 cm, working segment length =1 cm), a total of 61 patients with 87 thyroid benign nodules underwent ultrasound-guided radiofrequency ablation in our department from February 2013 to May 2014.The maximum diameter , volume, and volume-reduction ratio (VRR) were measured by ultrasound at 1, 3, 6, and 12 months after operation. Results The 61 patients with 87 thyroid benign nodules were successfully treated with radiofrequency ablation , including 3 cases receiving secondary ablation after 1 month.The maximum diameter of thyroid nodules was (2.24 ±0.63) cm preoperatively, and (1.82 ±0.81) cm, (1.52 ±0.72) cm, (1.03 ±0.43) cm, and (0.56 ±0.36) cm at 1, 3, 6, and 12 months after ablation, respectively.The maximun volume was (5.37 ±1.85) ml preoperatively, and (4.41 ±2.16) ml, (3.19 ±1.92) ml, (1.34 ±0.61) ml, and (0.69 ±0.41) ml at 1, 3, 6, and 12 months after ablation, respectively.The VRR were (20.7 ±13.2)%, (48.6 ±16.1)%, (77.3 ±4.7)%and (88.7 ±5.6)%, respectively at 1,3,6, and 12 months after ablation.At the final follow-up, 51 (58.6%) nodules disappeared completely. Conclusion Ultrasound guided radiofrequency ablation for thyroid benign nodules is safe and effective .

Objective To compare the therapeutic effects of closed reduction and transverse fixation with Kirschner wires and open reduction and fixation with mini-plates or Kirschner wires for the fifth metacarpal base fracture.Methods Data of 50 patients,who had been treated in our hospital for the fifth metacarpal base fracture from August 2005 to May 2012,were enrolled in this study.Twenty-six patients underwent closed closed reduction and transverse fixation with Kirschner wires,including 22 males and 4 females,aged from 17 to 41 years; according to the OTA classification,there were 8 of type A1,11 of type B1,and 7 of type C1.Twenty-four patients underwent open reduction and fixation with mini-plates or Kirschner wires,including 21 males and 3 females,aged from 19 to 46 years (average,26 years); according to the OTA classification,there were 11 of type A1,8 of type B1 and 5 of type C1.The total active motion (TAM) score of Chinese Medical Associaition was used to evaluate the range of motion of the joint.Results In closed reduction group,the operation time ranged from 14 to 33 min; the treatment costs ranged from 2018 to 2995 yuan; 23 patients were followed up for 6 to 26 months; X-rays showed the anatomic reduction was achieved in all patients,and the fracture healing time ranged from 4 to 7 weeks; according to the TAM score,the result was excellent in 15,good in 6 and fair in 2,and the excellent and good rate was 91.3％.In open reduction group,the operation time ranged from 45 to 105 min; the treatment costs ranged from 3874 to 4793 yuan; 22 patients were followed up for 6 to 21 months; X-rays showed the anatomic reduction was achieved in all patients,and the fracture healing time ranged from 4 to 8 weeks; according to the TAM score,the result was excellent in 13,good in 7 and fair in 2,and the excellent and good rate was 90.9％.Conclusion Closed reduction and transverse fixation with Kirschner wires is an effective method for the fifth metacarpal base fracture,which has several advantages,such as simple operation,stable fixation,satisfactory results,short operation time and low treatment costs.

BACKGROUND: During conventional treatment for bone tuberculosis, there is a low effective concentration of anti-tuberculosis drugs, and the therapeutic effect is poor. OBJECTIVE:To develop a new biomaterial as a slow-release artificial carrier that can be directly implanted into the surrounding tissue of bone tuberculosis, maintain a certain anti-tuberculosis drug concentration for a long time, thereby playing an effective therapeutic action. METHODS:Rifampicin/polylactic acid/glycolic acid microspheres and isoniazid/polylactic acid/glycolic acid microspheres were prepared using the emulsion-solvent evaporation method. Usingα-cyanoacrylate, a biological adhesive, two kinds of microspheres were processed into a long-term slow-release bicomponent drug carrier. Then, in vitro release characteristics of the dual-drug sustained-release carrier were observed. After that, the dual-drug sustained-release carrier was implanted into rabbit intertrochanteric femur bone defects for observing drug release concentrations, histocompatibility and bone defect healing at different time points after drug delivery carrier implantation. RESULTS AND CONCLUSION:For rifampicin/polylactic acid/glycolic acid microspheres, the mean particle size was (240±13)μm, and the drug loading load rate was (26±1.5)%. For isoniazid/polylactic acid/glycolic acid microspheres, the mean particle size was (250±10)μm, and drug loading rate was (28±1.8)%. The in vitro cumulative release rate could reach 80%for rifampicin and 90%for isoniazid at day 90. The in vivo released concentration of rifampicin and isoniazid within 90 days was (0.5±0.4) and (0.6±0.3)μg/g, respectively. There were a smal amount of infiltrated neutrophils between the fascia and muscle fibers after the drug delivery carrier was implanted, and the amount of neutrophils in the muscle were reduced significantly at day 59. X-ray plain film showed that bone defects decreased obviously in size. These findings indicate that this dual-drug sustained-release carrier can maintain a certain anti-tuberculosis drug concentration in the surrounding tissues of bone tuberculosis, which is expected to provide a new type of dual-drug delivery carrier in the surgical treatment of bone tuberculosis.

OBJECTIVETo investigate the application of the dorsal metacarpal perforator sliding flap for web-space reconstruction in congenital syndactyly.

METHODSAccording to the size and shape of skin defect at the web space after division operation of syndactyly, the corresponding intermetacarpal perforator sliding flap was designed. The edge of the flap was cut off, but its underlying tissue was not dissected. From May 2007 to November 2012, 28 web-spaces in 15 patients with syndactyly (10 male and 5 female) were reconstructed.

RESULTSAll the 28 flaps survived completely. The flap size ranged from 3 cm x 2 cm to 1.5 cm x 1.0 cm. 14 cases with 26 flaps were followed up for 10-22 months (average, 14.5 month). The reconstructed web spaces had normal appearance and movement range. The 2-point discrimination distance was 9-13 mm (average, 11 mm). According to the Swanson Standard, 18 fingers were graded as excellent, 8 as good and 2 as fair (excellent and good, 92.6%, 26/28).

CONCLUSIONSReconstruction of web-space in syndactyly with the dorsal metacarpal perforator flap has the advantages of easy handling, good cosmetic and functional results.

Objective To investigate the effect of dorsal digital island advancement flaps accompanied with digital arteries in reconstruction of finger pulp defect.Methods The study enrolled 36 cases (43 fingers) of pulp soft tissue defect treated from October 2007 to July 2011.There were 25 males (30 fingers) and 1l females(13 fingers),at age of 12-62 years (mean 34.5 years).Causes of injury included crushing in 19 cases (16 fingers),squeezing in 13 (16 fingers) and cutting in 4 (5 fingers).The fingers involved were the index (11),the middle (18),the ring (9),and the little (5).All wounds revealed the exposed bone with defect area ranging from 1.2 cm × 1.0 cm to 2.6 cm × 2.0 cm.Dorsal digital island flaps accompanied with digital artery were used for finger reconstruction and sensation was restored by anastomosing to dorsal branch of digital nerve.Donor sites were closed using skin grafts.Results All flaps survived.Thirty-three cases (39 fingers) were followed up for 5-26 months (mean 14.5 months).The flaps revealed satisfactory appearance,soft texture and good elasticity.The injured fingers were pain-free.Two point discrimination was 5-8 mm (mean 6.7 mm).According to the upper limb function criteria formulated by Hand Surgery Branch of Chinese Medical Association,the results were excellent in 24 fingers,good in 13 fingers,and fair in 2 fingers,with the excellence rate of 95％.Conclusion Dorsal digital advancement island flap vascularized with digital artery is relatively the best for finger pulp coverage,for the procedure is easy and safe,with satisfactory result.