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]To observe the clinical results of a hollow compression nail applied to treat the separation of elements of neural arch.[Method]Since February,2003,sixteen cases of separation of elements of neural arch have been treated with a hollow compression nail and bone grafting .The mean age of the group was 32,from 18 to 54.Five cases of separation of elements of neural arch located at L4 and the other 11 at L5.All the patients suffered from severe back pain.During the operation,the presence of the pars interarticularis defect was confirmed.The defects were cleared of any fibrous and cartilaginous tissues.The sclerotic bone margins were curetted until bleeding was noted .Autograft harvested from posterior ilium was done.The hollow compression nail was placed onto the index vertebra.[Result]All the patients were followed-up for more than 14 months (14～26 months).The significant back pain relief was found in all the cases.The bony healing was comfirmed in 11 cases,and the other 5 were in the process of bone growth.[Conclusion]The separation of elements of neural arch can be treated by the hollow compression nail without spinal fusion.The hollow compression nail is proved to be safe and effective.

Objective To explore the effects of single rectangle cage interbody fusion combined with pedicle screw fixa?tion for isthmic spondylolisthesis. Methods Data of the 34 cases with isthmic spondylolisthesis hospitalized at our department from March 2012 to February 2015 were retrospectively analyzed. All cases were operated by posterior pedicle screw fixation com?bined with single rectangle cage interbody fusion. There were 21 males and 13 females, aging from 18 to 63 years old (with an aver?age age of 41.6±8.2 years). Lesion segments were L4 in 15 cases and L5 in 19 cases;there were 12 cases in Meyerding I degree, 16 in II degree and 6 in III degree;28 patients had lower back pain associated with lower limb pain;19 patients had lower back pain associated with numbness of lower limb;18 patients had lower back pain associated with lower limb muscle weakness and 2 pa?tients had urination and defecation function disturbance. The surgical operation time, bleeding, postoperative drainage flow, graft fusion rate, complications were measured, and the intervertebral disc height, Japanese Orthopaedic Association (JOA), Oswestry disability indes (ODI), visual analogue scale (VAS) score were compared before and after treatment. Results The operation of 34 patients was successfully completed with the average operation time of 1.8 ± 0.8 h, and the bleeding in operation was 362.4 ± 93.7 ml, while postoperative drainage flow was 116.3±54.2 ml. All patients were effectively followed up for 12 months to 46 months (av?erage, one year and 10 months). Two weeks after operation, the intervertebral disc height were obviously increased (9.85±1.11 mm, 9.63 ± 1.04 mm), and there were significant difference compared with preoperative results (6.53 ± 0.98 mm, 6.24 ± 1.07 mm), but there were no significant difference compared with the latest follow?up results (9.23±1.33 mm, 9.30±1.16 mm). At the latest follow?up, JOA score was 13.19±1.08, ODI score 12.34±7.52, VAS score 2.23±1.51, and there were significantly different in each score compared with preoperative results. The symptoms of low back pain, lower limb nerve dysfunction and quality of life were signifi?cantly improved compared with preoperative situation. At the 16 month follow?up, all postoperative patients with vertebral body bone graft were judged to be fused. There was no pedicle screw loosening, fracture and so on. Conclusion Single rectangle cage interbody fusion combined with pedicle screw fixation for the treatment of isthmic spondylolisthesis is a kind of mini?invasive, less bleeding, safe and effective surgical method.

Objective To study the effect of early rehabilitation intervention on neurological deficits,equilibrium function, motor function and activities of daily life abilities in stroke patients. Methods Sixty two patients with acute stroke were allocated randomly into two groups: rehabilitation group and control group. The patients in the rehabilitation group were treated with a comprehensive rehabilitation regimen including the clinical treatment,Bobath technique, PNF technique and neuromuscular stimulation therapy. The patients in the control group were treated with medications. Neurological deficits,equilibrium function,motor function and ADL of all the patients were assessed before and after 28 days of rehabilitation intervention. Results The neurological deficit scores were reduced, while equilibrium function and motor function and ADL were increased after therapy in the two groups. The difference was significant ( P

Objective To investigate the efficacy of high frequency jet ventilation via the nasopha?ryngeal catheter in assissting ventilation in patients with obstructive sleep apnea hypopnea syndrome ( OS?AHS) undergoing gastroscopy. Methods Eighty patients with OSAHS of both sexes, aged 40-64 yr, weighing 65-99 kg, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with an apnea?hy?popnea index 20-40 events∕h, scheduled for elective painless gastroscopy, were divided into control group and test group using a random number table, with 40 patients in each group. In control group, a nasopha?ryngeal catheter 6.0-7.0 mm in internal diameter was inserted, and oxygen was inhaled at 4 L∕min through the catheter. In test group, a nasopharyngeal catheter 4.0 mm in internal diameter was inserted, and a high frequency jet ventilator was connected ( inspiratory∕expiratory ratio 1. 0 ∶ 1. 5, frequency 150 bpm, peak pressure 0.4 kPa, tidal volume 180 ml) . Anesthesia was maintained with propofol in both groups. The oc?currence of hypoxemia during ventilation, and peak value of partial pressure of end?tidal CO2 before induc?tion of anesthesia and during ventilation, and occurrence of chin lift, mask ventilation, and epistaxis after insertion of the catheter during operation were recorded. Results Compared with control group, the inci?dence of hypoxemia, peak value of partial pressure of end?tidal CO2 during ventilation, and incidence of chin lift, mask ventilation and epistaxis during operation were significantly decreased in test group ( P<0.05) . Conclusion High frequency jet ventilation via the nasopharyngeal catheter can be safely and effec?tively used to assisst ventilation in patients with OSAHS undergoing gastroscopy.

Objective To study the relationship between the expression level of PLK1 in castration-resistant prostate cancer (CRPC) tissues, and its relationship with pathological features. Methods Forty-four CRPC specimens including 28 samples from patients with prostate adenocarcinoma, 14 samples from patients with neuroendocrine prostate cancer (NEPC) and 2 samples from patients with other types of prostate cancer, and 10 normal prostatic hyperplasia specimens were collected from January 2010 to September 2016 in the Second Hospital of Tianjin Medical University. The expression levels of PLK1 in these tissues were detected by S-P immunohistochemistry. The relationship between PLK1 expression and pathologic factors was discussed. Results The positive expression of PLK1 was located in cytoplasm of carcinoma cells, and no express of PLK1 was found in benign prostatic hyperplasia tissues. The expression levels of PLK1 showed no significantly differences between different groups of age, local tumor invasion and regional nodal status, and the level of prostate-specific antigen (PSA, P>0.05). The expression level of PLK1 in patients with Gleason score>8 was higher than that in patients with Gleason score≤8. The PLK1 expression level was positively correlated with Gleason score (rs=0.441,P<0.05). Conclusion PLK1 protein is over-expressed in CRPC tissues, which can reflect the proliferation and differentiation of cancer cells and may be a potential marker of CRPC.

Objective To summarize the characteristics of medical service patterns at Grenada station of Mission Harmony-2015 so as to provide reference for future overseas services in similar island countries .Methods After detailed analysis of actual conditions of the countries visited and efficient communication with host country health organizations , we made an appropriate adjustment of the medical service pattern , which involved multi-site screening of general patients , followed by intensive checkups and treatment aboard the hospital ship .Results The procedure of diagnosis and treatment was streamlined and the efficacy of medical services was significantly enhanced to meet the maximum medical demand of local residents.Grenada became a station with the largest number of patients who received top-quality fee-free healthcare during Mission Harmony-2015 .Conclusion Adjustment of medical service patterns according to the actual conditions could deliver a desired effect during overseas missions .

Objective To systematically evaluate the efficacy and safety of 125I seed implantation for treatment of advanced pancreatic cancer.Methods An electronic literatuire search was performed about randomized controlled trials(RCTs) of 125I implantation for treamtent of advanced pancreatic cancer in CNKI,Wanfang Data,CBM,Cochrane Library,PubMed and Embase (from the date of building the database to November 2016).Two investigators independently screened literature,extracted data and assessed the risk bias of included studies,and the Meta-analysis was performed by using Revman 5.3software.Results There were 12 RCTs (n =689) included.Meta-analysis showed that the objective respond rate(ORR) (OR =3.24,95％CI2.33-4.52,P＜0.001),the 6-month survival rate(OR =3.61,95％ CI 1.53-8.52,P =0.003),the 12-month survival rate(OR =4.80,95％ CI 2.40-9.57,P ＜ 0.001) and the relief rate of pain were higher than those in the control group.However,there were no significant differences between both groups in the 2-year survival rate and the adverse reaction rate,which were (OR=2.36,95％ CI 0.47-11.74,P =0.29) and (OR =4.94,95％ CI 1.05-23.23,P =0.04),respectively.Conclusions The limited current evidence showed that 125I implantation for treatment of advanced pancreatic cancer is effective and safety.125I implantation can improve the ORR,short-time survival rate and pain relief rate.In addition,there was no significant increase in the incidence of related adverse events except for seed malposition.Although the quality and quantity of evidences is limited,it merits further study to provide high quality evidences.

Objective To evaluate the clinical effectiveness and safety of stents loaded with 125I seeds compared to conventional stents.Methods Literatures were searched in PubMed,EMbase,Cochrane Library,CBM,CNKI,Wanfang Data and other electronic databases from inception to November 2016.Two reviewers independently screened the literature according to the inclusion and exclusion criteria,extracted data and assessed quality of the included studies independently.Meta-analyses were performed using RevMan 5.3.Results A total of five RCTs and 14 CCTs involving 1 211 patients were included.The mean survival time of the 125I stent group was significantly higher than that of the control group [mean difference =4.11,95％ CI (2.16-6.07)P ＜0.001].The incidence of restenosis after 3:The available data showed that the incidence of re-staging of 125I stent in the treatment group was lower than that of the normal stent group [RR =0.23,95％ CI(0.12-0.62),P =0.002].Postoperative bleeding [RR =0.80,95％CI (0.52-1.23),P=0.30];Postoperative pain[RR=1.06,95％CI(90.88-1.27),P=0.55];postoperative stent shift [RR =0.53,95％ CI(0.27-1.05),P =0.07].The difference of incidence of complications was not statistically significant.There was no difference in the incidence of complications between the two groups.Conclusions The available data suggest that 125I stent is superior to common stent in the treatment of advanced esophageal cancer.There are no differences found in the incidence of complications between 125I stent and conventional stent.However,due to the limited quality of the included studies,more high-quality and multicenter-based studies are needed to verify the above conclusion.

Objective To explore the safety,effectivity and dosimetric continence of 3D-printing coplanar template(3D-PCT)combined with CT-guided 125I seed implantation in the treatment of non-small cell lung cancer(NSCLC).Methods From May 2014 to November 2016,a total of 20 NSCLC patients who were suitable for 125I seed implantation were recruited in this study.Of all the patients,10 received 125I seed implantation treatment by CT-guided combined with 3D-PCT (3D group),and the rest,by freehand puncture combined with CT-guided 125I seed implantation (free-hand group).During two days before the surgery,the patients received the CT scan.Then the digital imaging and communications in medicine (DICOM) was collected to input to the Brachytherapy Treatment Planning System (BTPS).The dose parameters including D90,D100,V100,V150,conformal index(CI),external index(EI),and homogneneity index(HI) were compared between pre-operation and post-operation.Pair t-test and single sample t-test were performed.Results V150 in 3D group between preoperation and postoperation showed statistically significant difference (t =-2.916,P ＜ 0.05),and there was no significant difference in the rest parameters(P ＞ 0.05).However,the number of seeds,V100,EI,HI in free-hand group between preoperation and postoperation showed statistically significant difference (t =-2.516,2.492,4.725,7.258,P ＜0.05),and there was no significant difference in the rest parameters(P ＞0.05).Comparison of indicuted that there was significant difference in V100,V150,CI,EI,HI and dose error rate between the 3D group and the free-hand group with single sample t test,the result showed V100,CI,EI,HI between two groups(t =2.598,2.278,4.637,4.616,-4.785,P ＜0.05),and there was no significant difference in V150 (P ＞ 0.05).Conclusions CT-guided 125I seed implantation brachytherapy combined with 3D-PCT for treatment of NSCLC safe is feasible,and dose controllable,and there is a certain advantage in the spatial distribution of seed compared with free-hand puncture.