Objective: To clone the Chinese MxA gene and to construct the recombinant adenovirus plasmid carring human MxA gene for further study.Methods: The peripheral blood mononuclear cells(PBMCs) were isolated by gradient fractionation,then total RNA was extracted from the PBMCs.The MxA gene was amplified by RT-PCR using the primers based on the published sequence of MxA.The PCR product,double-digested with restrictive endonucleases NotI and XbaI,was cloned into pAdTrack-CMV by molecular cloning technique.The plasimid pAdTrack-MxA was linearized with Pme I,and subsequently co-transformed into electrocompetent BJ5183 cells with an adenoviral backbone plasmid(pAdEasy-1).Recombinants were selected for kanamycin resistence and confirmed by PacI.Finally,the linearized recombinant plasmid was transfected into HEK 293 cells.Recombinant adenoviruses were generated within 7 to 12 days,then viral tite was checked by GFP.Results:Sequencing analysis revealed that the amino acid sequence of MxA gene had only one amino acid residue displace from isoleucine to valine(compared with that of published sequence) although there were five variations in nucleotide sequence.The restrictive endonuclease analysis confirmed that correct recombinant adenovirus plasmid was constructed.24 hours after transfection,the fluorescence was observed in 293 cells,and viral title checked by GFP was 1.59?108(pfu/mL).Conclusion: The MxA gene is cloned from a Chinese donor and a recombinant adenovirus vector containing human MxA gene is constructed successfully.It lays the foundation for further study of gene therapy for HBV.

BACKGROUND:Lumbar pedicle screw placement technique is a very mature technology in spinal surgery.There are many methods for determining the screw entry points and direction in the clinic.However,for patients with severe lumbar facet degeneration,severe osteophytosis can be found in the surgical area.Smooth and accurate screw placement has some difficulties.OBJECTIVE:To observe the effects of screw placement simulation using Mimics software on pedicle screw fixation before repair of severe lumbar facet joint degeneration.METHODS:A total of 78 cases of severe lumbar zygapophysial degeneration evaluated by CT and MR,who received single-segment lumbar pedicle screw fixation,were enroled in this study.They were randomly dividedinto two groups,including simulation screw group (n=39) and control group (n=39).Totaly 312 pedicle screws were implanted in the two groups.In the simulation screw group,Mimics software was used to perform 3D reconstruction before surgery and to simulate vertebral pedicle screw operation.In the 3D images,screw entry points and surrounding bony landmarks of pedicle screw were observed.The angle and depth of screw placement were measured.During the operation,pedicle screw placement was conducted in accordance with above results of observation and measurement.In the control group,pedicle screw was implanted according to the conventional method.After treatment,an X-ray CT examination was performed in the evaluation of the accuracy of pedicle screw placement.RESULTS AND CONCLUSION:No postoperative complication occurred in the simulation screw group.153 of 156 screws were located in vertebral pedicle in the postoperative examination.The correct rate of screw placement was significantly higher in the simulation screw group than in the control group (98.1％,88.5％,χ2=11.49,P＜0.05).The operation time,amount of bleeding and radiation exposure time were significantly shorter in the simulation screw group than in the control group (P＜0.05).These findings indicate that the lumbar facet joint degeneration accounted for a large proportion in patients with lumbar pedicle screw fixation.Facet joint degeneration increases the difficulties on distinguishing anatomical landmarks of insertion point,which brings a great difficulty for screw placement.The simulation of Mimics software in screw placement can improve accuracy in pedicle screw fixation in patients with severe lumbar facet joint degeneration,reduce complications,operation time,amount of bleeding,and radiation exposure time.

Objective The changes of CECs,EPCs and CPCs in the old type 2 diabetes mellitus retinopathy(DR)patients and the clinical purpose were discussed.Methods 30 cases of T2DM(including 10 DM cases without complications and 20 DM cases with retinopathy)and 10 cases of health controls were enrolled in this study.Results(1)The changes of CECs in DR Patients were significantly higher while the levels of EPCs and CPCs were significantly lower than those in the patients of the control group.(2)There were some corrlation beteen the level of CECs,EPCs and CPCs and the degree of diabetic retinopathy.(3)The serum EPCs levels in type 2 diabetics were negativey associated with body mass index,waist-hip ratio,abdominal circumference,FPG,triglyceride,UAER,creatinine.Conclusion The levels of EPCs and CPCs in T2DM-DR patients may be the important detected targets for early discovery and monitoring of DR.

Objective To design Radio-Frequency Identification(RFID) reader with multi-antennas of different spatial location.Methods Design scheme of RFID reader with multi-antennas of difference spatial location was presented,i.e.a separated setting with a reader and multiple detected antennas was adopted.Each antenna only identified tags in its own area.The receiving information from tags contains antenna code and data in tag's IC.Hence the reader can point the tags position through different antenna code.Results The application of RFID with multi-antennas of different spatial location to management system of combat readiness medicine can provide safe,exact and real-time information collection,processing and query for combat readiness medicine.Conclusion The system can not only provide location identification,but also reduce system cost.It will have wide application in future.

OBJECTIVE To investigate the drug-resistance of Enterobacter cloacae(ECL) and its carrying rate of gene encoding aminoglycosides modifying enzymes(AMEs).METHODS Totally 132 clinically isolated ECL strains were identified with VITEK-32.Their drug susceptibility was tested with K-B disc diffusion.The presence of ESBLs and AmpC was tested with three dimensional test.The genes encoding AMEs were detected with PCR and confirmed by sequence.RESULTS All strains of ECL were susceptible to IMP and MER.The resistance rate to AMK,LVX,FEP,CFS and CIP was 21.2%,35.6%,37.9%,46.2% and 48.5%,respectively with gradually increasing.Their resistance rate to other antibiotics ranged from 50.0% to 87.7%.Among 132 strains of ECL,45 strains producing ESBLs,accounted for 34.1%,27 strains producing AmpC,accounted for 20.5%,11 strains producing ESBLs and AmpC,accounted for 8.3% and 49 strains producing neither ESBLs nor AmpC,accounted for 37.1%.Seven strains without positive detection of ESBLs were resistant to CTX and CAZ,but carried the genes of resistance to TEM and SHV.The detection rate of genes encoding AMEs was about 62.1%,among which subtypes of aac(6′)-Ⅰb dominated with detection rate of 51.5%.Meanwhile the detection rate of subtype aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and aph(3′)-Ⅵ was 15.9%,12.9%,18.9%,37.1%,6.8% and 9.8%,respectively.The detection rate of carrying more than two types of AMEs was about 89.0%.There were 5 strains of presenting phenotypic resistance without detection of genes encoding AMEs.CONCLUSIONS The clinically isolated ECL strains are not only severe resistant to drug and but also multi-drug resistant.The carrying rate of genes encoding ESBLs,AmpC and AMEs is higher than estimated which is the leading cause of resistance to antibiotics.

Objective The purpose of this study was to observe the effects of bumetanide on elderly type 2 diabetic patients with middle and advanced stages nephropathy. Methods Forty cases with diabetic nephropathy (DN) were divided into two groups:control group (20 cases) and treatment group (20 cases) . The control group accepted furosemide (20 mg, once a day) and the bumetanide was orally administrated at the dose of 1.0 mg, twice a day to the treatment group for 3 months. The assessment of transferrin (TRF) and urine microalbumin (UALB) was performed at the time points at the end of 1, 4, 8, 12 weeks after treatment. Results (1) The levels of BUN, SCr and UA after treatment were lower than before treatment, but there were no significant differences. (2) In the treatment group, the levels of TRF and UALB have decreased after treatment for 1 week, with no statistically significant. But at the end of 4, 8 and 12 weeks,the differences were statistically significant ( <0.05) . Especially, during the three months follow-up,the levels of TRF and UALB at 12 weeks have decreased obviously. Conclusion The therapeutic effect of bumetanide on DN might be better than that of furosemide.

Objective To evaluate the clinical efficacy and adverse reactions of alizarin combined with anti-tuberculosis therapy for multidrug resistant pulmonary tuberculosis (MDR-PTB).Methods A total of 200 confirmed MDR-PTB patients admitted in the Affiliated Hospital of Hangzhou Normal University during June 2013 and June 2015 were enrolled in the study.Patients were randomly divided into study group and control group (100 in each).Both groups were given standard anti -tuberculosis treatment for 8 months, and additional alizarin was given to study group .Chi-square test was used to assess the differences in clinical efficacy, sputum negative conversion rate, cavity closure and lesion absorption rate , as well as the incidence of adverse reactions between two groups ( including patients categorized according to TCM syndrome ). Results There were 39 markedly effective cases, 51 improved cases, 10 ineffective cases in study group, and 22 markedly effective cases, 35 improved cases, 43 ineffective cases in the control group.The total effective rate in study group was significantly higher than that in control group (90% vs.57%, χ2 =28.262, P <0.01).For patients with TCM syndrome differentiation as phlegm -heat stagnating lung and those with qi-stagnation induced blood-stasis, alizarin combination therapy had significantly higher total effective rate than standard anti -tuberculosis treatment (78.78% vs.63.33%, χ2 =7.187, P <0.05;95.74% vs.42.31%, χ2 =73.997, P <0.01), but the difference was not observed in patients with TCM syndrome differentiation as deficiency of qi and blood (95.00% vs.88.89%, χ2 =5.025, P >0.05). There was no significant difference in sputum negative conversion rate between two groups (76% vs.55%,χ2 =2.190, P >0.05).The cavity closure and lesion absorption rate in study group ( 91%) was significantly higher than that in the control group (54%,χ2 =38.294, P <0.01).The adverse reaction rate in study group was 27%, which was significantly lower than that in control group (66%, χ2 =30.570, P <0.01).Conclusion Alizarin in combination with standard anti -tuberculosis therapy can improve the clinical efficacy and reduce adverse reactions in treatment of MDR -PTB.

Object To establish the quality standard for Wuhu Powder. Methods Radix Angelicae Sinensis (RAS), Radix Angelicae Dahuricae (RAD), and Flos Carthami (FC) were determined qualitatively by TLC. The content of cimicifugoside and 5-O-methylvisammioside were determined by HPLC. Results The linear ranges of cimicifugoside and 5-O-methylvisammioside were 0.370 0—1.850 0 ?g and 0.191 0—0.955 0 ?g. The recoveries were 100.74% and 98.33%, RSD were 0.67% and 1.91% (n=5), respectively. Conclusion The method is simple, accurate, credibility, which can be used as the quality control standard of Wuhu Powder.

Objective To construct and identify norepinephrine ( NE) complete antigen for the preparation of high sensitive and specific anti-NE monoclonal antibody .Methods Glutaraldehyde ( GA) and 1-Ethyl-3-(3-Dimethylaminopropyl ) carbodiimide ( EDC) were used to cross-link NE with carrier pro-teins (BSA, OVA) for NE complete antigen preparation under conditions of pH 4.5 or pH9.0.Three assays including UV scanning , SDS-PAGE and FeCl3 color reaction were performed for identification of NE com-plete antigen.Serum antibody titers were evaluated in mice model induced by intraperitoneal immunization with NE complete antigen .Results NE complete antigens were successfully prepared as indicated by the three identification assays .The coupling ratio was significantly increased in a time-depended manner under the condition of pH9.0 in comparison to that in the condition of pH 4.5.Indirect ELISA results showed that , when coating antigens and serum antibodies were prepared with the same cross -linking method , the serum antibody titers were significantly higher than those with different methods .Conclusion Anti-NE antibodies were successfully prepared by immunizing mice with NE complete antigens .

Objective To explore the correlation between the expression of somatostatin (SS), gastrin (GAS), apoptosis index (AI),and p53 gene in colonic carcinoma. Methods The expression of GAS, SS, p53 and apoptosis cell were detected by immunohistochemistry (streptavidin-biotin-pero xidase complex, SABC) and in situ apoptosis detecting technic (TUNEL) . Results AI is higher in SS high and middle expression cases than in low expression cases(q=5.06,q=3.95,P