Objective To produce the recombinant NS3 protease of the hepatitis C virus in insect cells. Methods The gene of HCV serine proteinase domain which encodes 181 amino acids was inserted into pFastBacHTc and the recombinant plasmid pFBCNS3N was transformed into DH10Bac competent cells for transposition. After the recombinant bacmid had been determined to be correct by both blue-white colonies and PCR analysis, the isolated bacmid DNAs were transfected into Sf9 insect cells. The bacmid DNA was verified to replicate in insect cells and package into baculovirus particles via PCR and electronic microscope analysis. The insect cells infected by recombinant baculovirus were determined by SDS-PAGE and Western-blot assays. The recombinant protein was soluted in N-Lauryl Sarcosine Sodium (NLS) and purifed by metal-chelated-affinity chromatography, then the enzymatic activity was measured with the protein substrate NS5ab and the antigenicity of recombinant protease was determined by enzyme-linked immunoabsorbent assay. Results The HCV NS3 protease domain was expressed in insect cells at high level and it was partially resolved in NLS;Totally 0.2 mg recombinant serine proteinase domain with high purity was obtained by metal-chelated-affinity chromatography from 5?10 7 cells and both enzymatic activity and antigenicity of the protein were proven to be good. Conclusion The recombinant HCV NS3 protease expressed in insect cells is a membrane-binding protein with good enzymatic activity and antigenicity.

Objective: Our eyes are one of the most complicated physical organs. Many solutions have been put forward for hyperopia, myopia, aging eye and xenetnbole, which shows that great attention has been paid to illnesses related with cornea. Ophthalmic refractive surgery is one of the most advanced medical fields in ophthalmology, and laser treatment of myopia has been widely used around the world. With the matured myopia treatment, the treatment of hyperopia has increasingly become the focus of attention. However, there hasn't been the effective treatment of hyperemia, in particular, high hyperemia. Therefore, it is both academically and practically valuable to explore the laser treatment for hyperopia. Methods: In this paper, on the basis of Virtual Reality(VR) technology, a semiconductor laser thermal angioplasty framework for medical simulation system is established. In combination with the methods of object-oriented design and modular design, a simulation system is designed and developed. Results: The system performs the initial simulation functions of forecast, calculation and demonstration of semiconductor laser comeal thermal angioplasty in the treatment for hyperopia. Conclusion: The interactive simulation of the system modules can enable medical workers to experience and learn the actual surgical operations through the rehearsal process, and can also improve the modes, efficiency and quality of medical education and training, which has its practical value for the improvement of study of medicine. Corneal information database can help medical workers to facilitate clinical cornea information management With the deepening study, the functions can also be improved and with the increase in the data of the clinical cases and the improvement of the database, the forecast will be more accurate. The perfect interface for database、record and the property operation in the database have been developed in database management module of the system, which can easily manage and apply the comeal data.

Objective To investigate the stability of jinyin qingre oral liquid. Methods According to the stability test guidelines for pharmaceutical preparations,the appearance,pH and relative density of jinyin qingre oral liquid were detected, the contents of geniposide,chlorogenic acid and salvia acid B were determined by HPLC. The chromatographic conditions were as follows:AtlantisT3 column(150 mm×4. 6 mm,3 μm),mobile phase A:0. 05% phosphoric acid,mobile phase B:acetonitrle, gradient elution at the flow rate of 0. 8 mL · min-1 ,and column temperature at 35 ℃. The detection wavelength was set at 254 nm. Results The appearance,pH,and relative density of jinyin qingre oral liquid did not change significantly with the acceleration test,but the contents of geniposide,chlorogenic acid and salvia acid B decreased time dependently. Conclusion Light and high temperature could decrease the stability of the jinyin qingre oral liquid.

Objective To explore the clinical effect of Habib 4X bipolar radiofrequency ablation assisted laparoscopic partial nephrectomy without blockage of renal artery for treatment of renal cell carcinoma (RCC).MethodsBetween October 2010 and June 2011, 14 patients with exophytic RCC (T1N0M0) were treated with Laparoscopic Partial Nephrectomy with (LRA) or without (LPN) Habib 4X Bipolar Radiofrequency ablation assisted.The LRA group included 6 patients,and the maximum tumor diameter was 2.1 - 3.5 cm (mean,3.1 cm).The LPN group included 8 patients,and the maximum tumor diameter was 2.0 -3.7 cm (mean,3.0 cm).The operative time,average intra-operative blood loss,postoperative hospital stay and incidence of postoperative complications were recorded.Changes of hemoglobin (Hb),serum creatinine (SCr) and ipsilateral glomerular filtration rate (GFR) before and after operation were analyzed and compared. Results The 14 surgical procedures were successful without conversion to open operation.In LRA group,renal artery blocking was unnecessary,the Hb,SCr and GFR before and after operation were (127 ± 19) g/L vs.(124 ± 19) g/L、(96 ±39) μmol/L vs.(92 ±29) μmol/L and (42 ± 12) ml/min vs.(40 ± 13) ml/min,respectively,and the difference was not significant (P ＞0.05).In LPN group,the renal artery blocking time was 20-52 min (mean,32 min),the Hb,SCr and GFR before and after operations were (130±17) g/L vs.(112±15) g/L,(92 ±31) μ mol/L vs.(110±28)μmol/L and (43 ± 14) ml/min vs.(30 ± 11 ) ml/min respectively,and the difference was significant (P ＜0.05 ).The operative time was ( 86 ± 20) min and ( 112 ± 43 ) min,the estimated blood loss was (94 ± 18)ml and (256 ± 58) ml,and the postoperative hospital stay was (5.0 ± 0.8) d and (7.8 ± 1.2 ) d,respectively.The difference between the two groups was significant (P ＜ O.05).Complications such as bleeding,gross hematuria,and leakage of urine were noted in LRA group. Conclusions Habib 4X Bipolar Radiofrequency ablation assisted LPN without blockage of renal artery is a safe and effective intervention with a relatively low incidence of complications.And it is superior to LPN in operative time,intraoperative blood loss and postoperative recovery.

Objective To explore the clinical effect of percutaneous osteoplasty with SKy bone expander system in the treatment of calcaneal fracture.Methods 63 feet of 55 patients with calcaneal fractures were involved in this study.According to Sanders fracture classification including 38 feet of Sanders Ⅱ,18 feet of Sanders Ⅲ,7 feet of Sanders Ⅳ.There were 6 to 16 days interval between the injuries and the surgeries.The patients were treated by percutaneous osteoplasty with the SKy bone expander system.The standard of operation was the satisfaction of reduction and Bohler's and Gissane's angles under X-ray.Results All of 63 feet of 55 patients were followed up for average 22 months.According to the criterion of therapeutic effect,the results were as follows:excellent in 30 cases,good in 26 cases,fair in 7cases,and no poor case.The excellent and good rate was 88.9％.Conclusion Percutaneous osteoplasty with SKy bone expander system in the treatment of calcaneal fracture,especially in reduction and fixation of Sanders type Ⅱ and Sanders type Ⅲ,can recover Bohler's and Gissane's angles,significantly shorten the duration of illness,and has fast recovery and can possess satisfactory curative effect,and it is worth popularizing.

Objective To explore the prognostic value of modified Glasgow prognostic score (mGPS) and risk factors in predicting overall survival (OS) in the castrate-resistant prostate cancer (CRPC) treated with docetaxel-based chemotherapy.Methods We retrospectively analyzed the data of 48 consecutive Chinese patients with CRPC received docetaxel-based chemotherapy in our institution from January 2008 to January 2012.Patients were divided into three groups according to the mGPS:0,1 and 2 score groups,and compare the OS among the three groups.Variables that were influenced the efficacy of chemotherapy were included in the univariate analysis and multivariate model.Survival analysis was performed using Kaplan-Meier curves,and the differences in overall survival rates were assessed using the Logrank test.Results The follow-up was performed until April 2014.There were 48 CRPC patients including mGPS 0 score group 30 cases,mGPS 1 score group 11 cases and mGPS 2 score 7 cases.The median OS was 22,11,9 months,respectively,P＜0.01.Univariate analysis showed that:prechemotherapy baseline total prostate-specific antigen (tPSA),Eastern Cooperative Oncology Group (ECOG) score,the number of chemotherapy cycle,visceral metastasis and PSA response were associated with poor prognosis (P＜0.05).Multivariate analysis showed that:prechemotherapy mGPS 1-2 score,baseline tPSA＞60 μg/L,the number of cycles of chemotherapy≤5,with visceral metastasis and PSA response in patients with CRPC were independent risk factors for prognosis in the CRPC treated with docetaxel-based chemotherapy.Conclusion mGPS is an independent risk factor for prognosis in the CRPC patients treated with docetaxel-based chemotherapy.

Objective　To define the range of the normal values of MRI signals of hippocampal formation(HPF) for the diagnosis of early stage Alzheimer's disease and hippocampal sclerosis.Methods　MRI signals of 254 normal adults were measured on the transverse section.Results　HPF signal intensities were:T1 relaxation times,629±73 ms;T2 relaxation times,83±5.5 ms;and proton density value,5978±651.The T2 value range(χ±2S)was 72～94 ms,and mean 83 ms.T2 value greater than 104 ms will be associated with evidence of hippocampal pathologic changes.The mixed signal intensity of T2WI(30,60,120)were:3907±407,2657±347,1288±174.The signal intensity ratios of T2WI were:1.02,1.07,1.13.Conclusion　As for the histological features of temporal lobe cortex and HPF,T1relaxation times and proton density values are matched but T2 relaxation times of HPF are relatively longer.

Objective To investigate the incidence and risk factors of the complications in perioperative intra-aortic balloon pump (IABP) supported cardiac surgical patients. Methods The clinical data of adult cardiac surgery patients undergoing IABP in Fuwai Hospital from January 2005 to January 2017 were enrolled. The patients were divided into complications group and no complications group. Demographic characteristics, diagnosis, perioperative clinical parameters, IABP related data, and IABP complications (including ischemia, bleeding, vascular injury and mechanical problems) were collected. The incremental risk factors of complications related IABP were analyzed by logistic regression. Results During the 12-year period, 522 patients received IABP support, with 388 male and 134 female; the mean age was (61.79±9.35) years; the complications related to IABP occurred in 25 patients, and overall complication rate was 4.79%; 87 IABP patients were dead in-hospital, the overall mortality was 16.67%, no patient died due to complications. The complications rate was higher in the female patients (40.00% vs. 24.95%), and was more in patients with age ≥ 65 years old (80.00% vs. 38.03%), more with higher body mass index [BMI (kg/m2): 25.45±13.71 vs. 22.95±3.45], diabetes mellitus (44.00% vs. 26.76%), combination treatment with extra-corporeal membranous oxygenation (ECMO: 20.00% vs. 5.03%) and prolonged IABP support time (hours: 134.4±90.3 vs. 109.8±89.1, all P < 0.05). There was no significant difference in the incidence of complications among preoperative IABP support, intra-operative IABP support and post-operative IABP support [3.30% (3/91), 5.46% (10/183), 4.84% (12/248), χ 2 =0.629, P = 0.730]. Bleeding from puncture site occurred in 14 cases (2.68%) without severe bleeding. Limb ischemia occurred in 9 cases (1.72%). One patient (0.19%) was under another surgery because of retroperitoneal hemorrhage caused by vascular injury. One patient (0.19%) was unsuccessful due to a balloon leak. It was shown by logistic regression analysis that presence of age ≥ 65 years [odds ratio (OR) = 2.320, 95% confidence interval (95%CI) = 1.011-1.806, P = 0.047], diabetes mellitus (OR = 2.281, 95%CI = 1.016-5.120, P = 0.026) and combination treatment with ECMO (OR = 4.341, 95%CI = 1.240-15.196, P = 0.040) were found to be the risk factors of complications related to IABP. Conclusions IABP complication rates are generally low. The frequent complications during IABP support is bleeding from site of catheterization and limb ischemia. When patients were treated with IABP, those with older age, diabetes mellitus and combination with ECMO should be monitored closely in order to reduce complications.

Objective To investigate integration and expression of adeno-associated virus (AAV) vectors in neuronal PC12 cells,the result of which can be applied in further gene therapy of diseases of the central nervous system. Methods Human neurotrophin-3(hNT3)genes were inserted into AAV vectors. Then the recombinat AAV plasmids were encapsidated as recombinant virions. PC12 cells were transfected with the virions and the positive cells were selected by G418. The transfection positive (hNT3 modified)PC12 cells were cultured for several generations and the cellular genomic DNA and total RNA were extracted. We investigated the integration locus of AAV vectors by Southern blot and transcript situation of foreign genes by dot blot. Results The hybridization tests showed that AAV introduced foreign genes were stably integrated, but at random locus, and robustly transcribed in hNT3 modified PC12cells. Conclusion AAV vectors can serve as high efficiency vectors of target genes in neuronal PC12 cells.