Objective To express dengue virus(DENV)NS2B-NS3 protease in E.coli,optimize the expression conditions and determine the enzyme activity,so as to lay a foundation of screening and discovering of lead compounds targeting DENV.Methods Codon-optimized NS2B-NS3 gene was inserted into pET-28a vector to construct recombinant prokaryotic expression plasmid pET-28a-NS2B-NS3,which was transformed E.coli Rosetta(DE3)competent cells and induced by IPTG to express NS2B-NS3 protease. The optimal expression conditions of NS2B-NS3 protease in E.coli were determined by optimizing induction length,induction temperature and IPTG concentration. NS2B-NS3 protease was isolated and purified by HisTrap~(TM) affinity chromatography column and measured for the protease activity by fluorescence resonance energy transfer(FRET)assay.Results The recombinant prokaryotic expression plasmid pET-28a-NS2B-NS3 was constructed correctly as identified by restriction analysis(NheⅠ/XhoⅠ)and sequencing. The optimal expression conditions of NS2BNS3 protease in E.coli were as follows:induction temperature of 20 ℃,induction length of 10 h and IPTG concentration of0. 2 mmol/L. The purified NS2B-NS3 protease showed a purity of more than 90% with a exhibited a of 20 mg/L,which bound to mouse monoclonal antibody against His-tag specifically and had good hydrolytic activity with a specific activity of 16. 111 U/mg,a K_m of 16. 46 μmol/L and a k_(cat) of 0. 028/s.Conclusion DENV NS2B-NS3 protease with high purity and activity was successfully prepared,which laid an experimental foundation of the establishment of high-throughput screening model for inhibitors targeting NS2B-NS3 protease.

150 eyes (85 patients) of angle closure glaucoma treated with Nd: YAG laser iridectomy were analyzed to reveal that the basic factors leading to posterior synechiae were the dosage and duration of miotics used. In the cases with posterior synechiae, miotics of higher dosages were used before and after the laser iridectomy and also a longer duration before the operation than in cases without the sequelae. With posterior synechiae, the pathological damages to the optic nerve worsened with time to affect the visual acuity and the control of IOP, and also interfered with fundus examination and cataract extraction when needed. The study indicated that proper dosage of laser energy,intensive postoperative use of topical steroids, early dilatation of the pupil, and appropriate application of mioties were conducive to the prevention of posterior synechiae.

Objective To investigate medical and nursing interns' differences and similarities of role expectation on clinical nurses,and provide the basis for further carrying out physician-nurse cooperative education.Methods Using self-designed questionnaires of role expectation on clinical nurses to investigate medical and nursing interns of two degrees in Zhengzhou University who already had full ten months' internship,among them,154 medical students,145 nursing students.Results Medical and nursing students' differences were mainly on “physician-nurse collaboration and communication.Nursing students were more active than medical students.Both medical and nursing students had the lowest scores in item 15 (reverse entry)“The nurses just need to perform doctors' orders,without providing comments or suggestions”.Conclusions Colleges and clinical teachers should encourage nursing students to learn more medical knowledge,strengthen their clinical practice capacity,improve their professional competence and professionalism,thereby enhancing the status of nursing and nurses.Launching physician-nurse collaboration education in nursing and medical students,and help medical and nursing students realize nurses' different roles,improve the trust between doctors and nurses,promote the development of physician-nurse relationship.

The postoperative management of papillary thyroid microcarcinoma (PTMC) is one of the most important parts of management for PTMC,and it includes the initial management and long-term management of PTMC.The initial management of thyroid cancer is based on initial recurrence risk stratification system.The initial risk stratification system can be used to guide initial TSH suppression therapy,RAI assistant therapy and management of follow up.Initial risk assessment of recurrence,as a static representation of the patient in the first few weeks post-operatively,could not be continually updated during follow up.In order to account for the changes of the recurrence risk and disease specific mortality,re-stratification of risk should be performed during follow up,using all the clinical,biochemical,imaging (structural and functional),and cytopathologic findings obtained during follow up.Re-stratification of risk can inform clinicians on degree of ongoing TSH suppression,frequency and intensity of follow-up,and need for additional therapies were recommended in guideline.Adjustment of TSH suppression therapy should incorporate the impact of side effects and set TSH targets to ensure the benefits of TSH suppression outweigh against the potential risks.

Objective To investigate the level of urinary TGF-β1 and evaluate its clinical significance in patients with chronic nephritis.Methods 22 healthy people and 40 patients with chronic nephritis were selected,the levels of urinary TGF-β1 were measured by RT-PCR and ELISA.Results Urinary TGF-β1 were significantly increased in Mesangial Proliferative Glomerulonephritis (76.44± 24.29 ng/mmol.Cr vs 31.53 ± 11.59 ng/mmol.Cr,P<0.01);Focal Glomerular Sclerosis (65.07±20.59 ng/mmol.Cr vs 31.53±11.59 ng/mmol.Cr,P<0.01)and IgA Nephritis (67.01±21.95 ng/mmol.Cr vs 31.53±11.59 ng/mmol.Cr,P<0.01),comparing with normal group.Conclusion The levels of urinary TGF-β1 could be a predictor in reflecting the sate of renal diease,estimating pathological type,diagnosing the renal diease and observing the therapeutic effects.

Parkinson’ s disease ( PD) is a neurodegenerative disorder primarily characterized by resting tremor, muscular rigidity, akinesia and postural reflex impairment.Behavioral tests of PD in animal models are essential for under-standing the pathogenesis of PD as well as for the development and testing of potential therapeutics .Here we mainly use the 6-hydroxydopamine (6-OHDA)-induced rat model, to introduce a review on the research progress in non-drug-induced be-havioral tests of motor function in PD rats.

Objective To evaluate the feasibility of waist-to-height ratio (WHtR) to reflect the body composition of people aged 60-69 years in Hubei minority areas.Methods A total of 398 eligible residents between 60-69 years old were selected through a stratified sampling method.Body height,weight,waist circumference,hip circumference and skinfold thickness were measured at the interview.Bland-Altman analysis was applied to estimate the consistency among WHtR,body mass index (BMI),waist-to-hip ratio (WHR),and Suzuki-changling formula.Results After standardization,the mean of WHtR and BMI was 0.007,the mean of WHtR and WHR was 0.007,and the mean of WHtR and Suzuki-changling formula was 0.711,which revealed that there were good consistency among WHtR,BMI,WHR and Suzuki-changling formula.Conclusions WHtR may serve as an typical index to reflect the body composition of people aged 60-69 years in Hubei minority areas.

Carcinoma of unknown primary (CUP) is a diagnostic and therapeutic challenge in oncology.The early and correct detection of the primary tumors of CUP is very important to its diagnosis, treatment, therapeutic response evaluation and prognosis prediction.This review summarizes the implications and development of the imaging technologies, especially 18F-FDG PET/CT scan, in the diagnosis, treatment and prognosis prediction of CUP patients.

Objective The aim of this study is to investigate the effects of the initial amount of total RNA input,adapter dilution and PCR amplification cycles on the results of small RNA library preparation and high-throughput sequencing.Methods Based on the sequencing reads number,the number of detected miRNAs and the accuracy of quantitative detection,we explored the effects of initial Total RNA,dilution ratio and PCR cycles on the quality of microRNA library preparation and high-throughput sequencing,respectively.Results For libraries preparation of the normal RNA input(1 μg),the adapter dilution combined with 22 PCR cycles could gain the best quality of sequencing.In the low-input libraries(10 ng),adapter dilution and increased PCR cycles would also improve the quality of sequencing,but as compared with the 1 μg library,there was lower correlation with microarray quantitative results in general.Conclusions The initial amount of RNAs input has the biggest effects on the quality of sequencing,and the quantity accuracy rate of low-input libraries is lower than the normal libraries generally.Increasing the PCR amplification cycles properly is indispensable to low-input libraries.

4.0 mg/L) fluorosis areas in 2008.The fluoride content of the drinking water,the chlidren's dental fluorosis and urinary fluoride were investigated.Results The fluoride in drinking water in the water improved areas was(1.4?0.8)mg/L,which was significantly lower than that in the non-water improved areas(2.6?1.5)mg/L(P