Based on the gene sequence of AIV matrix protein 2(M2) and two cytotoxic T-lymphocyte epitopes derived from nucleoprotein,the prokaryotic expression vector pET-3M2e-NP1-NP2 was constructed.The target gene was expressed in the solvable form in BL21(DE3) when induced with 1.0 mmol/L IPTG and Western blot analysis showed that the expression product had good immunogencity.Purified fusion protein was mixed with various amount of adjuvant,including Freund's,Vash oil and chitosan,and then immunized 20-day-old chicken by intramuscular injection and boosted 3weeks later.Blood samples were collected weekly following the primary vaccination.The anti-M2e antibody was detected with ELISA method with the synthesized peptide as coated antigen;the neutralizing ability of anti-serum was evaluated on MDCK cell line and chick embryo,the CD4+ and CD8+ T lymphocyte amounts in peripheral blood of immunized chicken was measured by flow cytometry.Results showed that the fusion protein can induce immunological reaction,and the antibody can bind with the viral M2 protein that expressed on the surface of MDCK cells.Flow cytometry result showed that CD4+ and CD8+ T lymphocyte in peripheral blood increased obviously following immunization(P

M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

Objective To investigate the relationship between the fluorouracil pathway gene and effect of chemotherapy in advanced gastric cancer after surgery. Methods 52 postoperative patients with advanced gastric cancer using FOLFOX4 6-cycles combined chemotherapy were collected to set up the database. The expression of thymidine synthase (TS), dihydropyrimidine dehydrogenase (DPD) and orotate phosphoribosyltransferase(OPRT) in tumor tissue and adjacent non-tumor tissue of 52 patients with advanced gastric cancer were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The influence of fluorouracil pathway gene on chemotherapy was investigated. Results The TS-mRNA level in tumor tissue was significantly higher than non-tumor tissue (0.92±0.28 vs 0.71±0.30) (t = 3.730, P =0.001).OPRT-mRNA level in tumor tissue was positively correlated with the non-tumor tissue (r =0.45, P =0.001). No correlations were observed among other gene expressions. Patients whose high OPRT-mRNA gene expression in their tumors and non-tumor tissue showed an obviously better survival (t = 3.036, P =0.003;t = 2.713, P =0.009). Patients with low DPD-mRNA gene expression survived longer than those with high DPD-mRNA gene expression in tumor tissue with statistical significance(t = 2.708, P =0.009), whereas prolonged survival was observed in patients with high DPD-mRNA gene expression in non-tumor tissue (t = 2.616, P =0.012).Conclusion There is close relationships between chemotherapy in advanced gastric cancer and the expression of DPD-mRNA, OPRT-mRNA;while the expression of TS-mRNA have no relation with the survival time.

AlM: To observe the effect of anterior chamber depth and angle change after pterygium excision.METHODS:Thirty cases (30 eyes) of primary pterygium were underwent pterygium excision. Central anterior chamber depth, four direction angle open distance ( AOD ) and open angle ( AA ) were measured preoperatively and postoperatively by ultrasound biomicroscopy ( UBM ) and the intraocular pressure was observed.RESULTS:Preoperative and Postoperative intraocular pressure were 15. 17±10. 6 and 16. 23±2. 61mmHg, and the central anterior chamber depth were 2. 28±0. 39 and 2. 33± 0. 24mm. The four directions of AOD and AA were no statistical difference.CONCLUSlON:The anterior chamber depth and the angle change is not obvious after pterygium excision.

OBJECTIVETo study the effect of electro-acupuncture (EA) at points along Foot Yangming Channel on metabolite of ulcerative colitis (UC) rats' cerebral cortex and to identify key metabolites by referring to Pi/Wei-brain related theory in Chinese medicine (CM).

METHODSThe UC rat model was set up by dextran sulfate sodium (DSS) method. Male SD rats were randomly divided into the model group and the EA group, 13 in each group. Another 13 rats were recruited as the blank control group. Rats in the blank control group and the model group received no EA. EA was performed at Zusanli (ST36), Shangjuxu (ST37), and Tianshu (ST25) for 5 days by using disperse-dense wave. Then all rats were sacrificed. Their recto-colon and the ileocecal junction were pathomorphologically observed by light microscope and transmission electron microscope (TEM). Cerebral cortexes were extracted. Water-soluble and lipid-soluble brain tissue metabolites were respectively extracted for metabolic research using 1H nuclear magnetic resonance (1H-NMR).

RESULTSEA could obviously improve the general condition of UC model rats, decrease the value of DAI, reduce the infiltration of inflammatory cells in the intestinal tract, stabilize structures such as mitochondria, endoplasmic reticulum and so on (P <0.05). 1HNMR analysis showed that in the model group, contents of glutamic acid, cholesterol, very low density lipoproein (VLDL) in the pallium obviously decreased, while alanine and low density lipoprotein (LDL) significantly increased. After EA, levels of lactic acid, glutamic acid, total cholesterol (TC), and VLDL all increased, and levels of alanine and LDL decreased. All indices were approximate to those of the blank control group.

CONCLUSIONEA at Foot Yangming channel was found to have some effect on metabolites in the brain tissue of UC model rats, which had specific metabonomic material basis and mechanism based on the Pi/Wei-brain related theory.

Objective To investigate the ways of recovery guidance for aged patients with vertigo. Methods 450 aged patients with vertigo were divided into two groups. Patients in the experimental group were those coming to our ENT outpatient department in the odd months and patients in the control group were those coming to our ENT outpatient department in the even months. The 234 patients in the experimental group were treated with recovery training of standing balance training, head-moving, visual balance training and psychological counseling. The 216 patients in the control group weren't given any guidanco of balance training but medical treatment and psychological counseling. Evaluations were conducted after the 1st, 2nd, and 3rd months of treatment respectively. Results The relieving rate was 17.09%, 82.05% and 88.03% respectively in the experimental group and 2.78%, 26. 85% and 41.67% respectively in the control group after the 1st, 2rd, and 3th months of treatment(P < 0. 001 ). Condusions Standing balance training, head-moving, visual balance training and psychological counseling are effective in the treatment for aged patients with vertigo.

Objective To explore the clinical efficacy of ultrasound - guided injection of platelet-rich plasma (PRP) for the treatment of recalcitrant lateral epicondylitis. Methods From September 2014 to June 2016, a total of 15 patients with recalcitrant lateral epicondylitis received ultrasound - guided injection of autologous PRP therapy, including via left arm injection (n=2) and via right arm injection (n=13). By using twice centrifugal method, the patient' s own venous whole blood was centrifuged to obtain PRP. All patients underwent PRP injection once a week, a total of 3 treatments were performed for each patient. Results After the first injection of PRP, the patients were followed up for 12 months. One month after the treatment, visual analogue scale (VAS) score was obviously improved, at 3 months after the treatment the improvement of VAS score reached its peak and it remained at this level until 12 months after the treatment. The elbow joint function, which was evaluated with modified MAYO elbow score, was also significantly improved in one month after the treatment, and the clinical effect was sustained to 12 months after the treatment. Conclusion Ultrasound - guided precise injection of PRP can effectively improve the pain and the elbow joint function caused by recalcitrant lateral epicondylitis. (J Intervent Radiol, 2018, 27:238-241)

BACKGROUND: Studies have shown that bone marrow mesenchymal stem cell (BMSC) transplantation can effectively improve cardiac function after myocardial infarction. However, few reports have been issued on myocardial electrophysiology after BMSCs transplantation. OBJECTIVE: To observe the effects of BMSCs transplantation on voltage-gated K+channel protein and myocardial infarction-related cytokines, thereby providing basic evidence for further exploration on the mechanism underlying arrhythmia in myocardial infarction due to BMSCs transplantation. METHODS: Forty male Wistar rats, SPF grade, were randomly divided into four groups: sham group, model group, cell culture medium group and BMSCs group. The myocardial infarction model was created in rats by permanent ligation of the left descending coronary artery. At 15-20 minutes after surgery, BMSCs (100 μL, 1×106) or cell culture medium (100 μL) was injected at four sites in the peri-infarct zone. Four weeks after cell therapy, cardiac samples were taken, the pathological morphology of the infarcted myocardium was observed by hematoxylin-eosin staining, and the infarct size was calculated; the expression levels of voltage-gated K+channel proteins Kv1.2 and Kv1.5 and cardiac troponin T (cTnT) were measured by western blot assay; and the expression levels of apoptotic factor (Caspase-3), autophagy factor (Bcl-2), nitric oxide and superoxide dismutase were tested by immunohistochemistry. RESULTS AND CONCLUSION: Compared with the model group and cell culture medium group, the infarct size decreased in the BMSCs group (P < 0.05); the expression levels of cTnT, Kv1.5 and superoxide dismutase increased (P < 0.05), and the expression levels of Caspase-3, Bcl-2 and Kv1.2 decreased (P < 0.05) in the BMSCs group. In summary, BMSCs transplantation can promote the expression of voltage-gated K+channel proteins, and improve anti-oxidation capacity of the myocardium and decrease apoptosis and autophagy.

Objective To evaluate the value of transesophageal echocardiography (TEE) in transthoracic minimally invasive device closure of ventricular septal defect(VSD).Methods A total of 164 cases of VSD were recruited as candidates to receive transthoracic minimally invasive device closure between January 2007 and October 2010,including 138 perimembranous VSDs,3 muscular VSDs and 23 supracristal VSDs.Among these groups,85 male patients were included.Four-champer view,five-champer view,left ventricular long-axis view,short-axis view and right ventricular inflow view were detected to evaluate the availability of device closure.Results A total of 152 cases(92.7%)were successfully closed with a device.All the patients were followed up more than 3 months arranged with a standard protocol.No complete atrioventricular block (CAVB) or associated valvular complications were observed.Three of the five cases with traced residual shunt after device closure closed spontaneously.Conclusion TEE plays an important role in transthoracic minimally invasive device closure of ventricular septal defect,which has been proved by the good follow-up results without CAVB and associated valvular complications.

To construct a recombinant T7 phage expressing matrix protein 2 ectodomain (M2e) peptides of avian influenza A virus and test immunological and protective efficacy in the immunized SPF chickens. M2e gene sequence was obtained from Genbank and two copies of M2e gene were artificially synthesised, the M2e gene was then cloned into the T7 select 415-1b phage in the multiple cloning sites to construct the recombinant phage T7-M2e. The positive recombinant phage was identified by PCR and sequencing, and the expression of surface fusion protein was confirmed by SDS-PAGE and Western-blot. SPF chickens were subcutaneously injected with 1 X 10(10) pfu phage T7-M2e, sera samples were collected pre- and post-vaccination, and were tested for anti-M2e antibody by ELISA. The binding capacity of serum to virus was also examined by indirect immunofluorescence assay in virus- infected CEF. The immunized chickens were challenged with 200 EID50 of H9 type avian influenza virus and viral isolation rate was calculated to evaluate the immune protective efficacy. A recombinant T7 phage was obtained displaying M2e peptides of avian influenza A virus, and the fusion protein had favorable immunoreactivity. All chickens developed a certain amount of anti-M2e antibody which could specially bind to the viral particles. In addition, the protection efficacy of phage T7-M2e vaccine against H9 type avian influenza viruses was 4/5 (80%). These results indicate that the recombinant T7 phage displaying M2e peptides of avian influenza A virus has a great potential to be developed into a novel vaccine for the prevention of avian influenza infection.
Animals ; Antibodies, Viral ; blood ; Bacteriophage T7 ; genetics ; immunology ; metabolism ; Chickens ; Enzyme-Linked Immunosorbent Assay ; Gene Expression Regulation, Viral ; Immunization ; Influenza A virus ; genetics ; immunology ; Influenza Vaccines ; immunology ; Influenza in Birds ; immunology ; metabolism ; prevention & control ; Peptides ; genetics ; immunology ; metabolism ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; Specific Pathogen-Free Organisms ; Viral Matrix Proteins ; genetics ; immunology ; metabolism