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.

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.

Acupuncture Points ; Animals ; Cerebral Cortex ; metabolism ; Colitis, Ulcerative ; Electroacupuncture ; Lipids ; Male ; Rats, Sprague-Dawley

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.

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

BACKGROUNDGrowth and development of infants has been an important topic in pediatrics for a long time. Infants must be provided with food containing all necessary nutrients. Breast milk is believed to be the most desirable natural and cheapest food for well-balanced nutrition. But with the progress in the development of substitute food in developed countries, it is thought that formula milk can meet the requirement for infant growth. During early infancy, growth, as the most sensitive index of health, is therefore a critical component in evaluating the adequacy of breast-feeding, mixed-feeding and formula feeding. Iron status is another important index of infant health. Iron deficiency anemia remains the most prevalent nutritional deficiency index in infants worldwide. This study is to compare infants in Beijing at 4 months who are on three different feeding modes (breast feeding, mixed feeding and formula feeding) in physical changes and iron status. The results may provide new mothers with support in feeding mode selection, which will also be helpful to the China Nutrition Association in feeding mode education.

METHODSThis is a cohort study. One thousand and one normal Beijing infants were followed regularly for 12 months. Body weight and horizontal length were measured. Hemoglobin, red blood cell counts, mean corpuscular volume, mean corpuscular hemoglobin and serum iron were analyzed at 4 months.

RESULTSThe breast feeding percentage in the first 4 months was 47.9%. The feeding mode was not significantly related to maternal delivery age, education, labor pathway nor infant sex (P>0.05). Infant boys and girls exclusively breast-fed from 0 to 4 months had the highest weight at 0-6 months. The anemia rate of breast-fed infant boys at 4 months was the highest.

CONCLUSIONSBreast feeding should be given more emphasis. It is compulsory for new mothers to breast-feed their infants if possible. Social environment should also guarantee the requirement for breast feeding. Furthermore the normal values of hemoglobin, mean corpuscular volume and serum iron, which were originally used to judge children's iron deficiency anemia, might not be optimal for evaluating infants. There might be a need to develop sex-specific cutoff levels of hemoglobin, mean corpuscular volume and serum iron for infants.

Anemia, Iron-Deficiency ; epidemiology ; Breast Feeding ; Child Development ; Erythrocyte Indices ; Hemoglobins ; analysis ; Humans ; Infant ; Infant Formula ; Infant, Newborn ; Iron ; blood ; Prevalence

Little is known about how gap junctions are involved in the respiratory-related or other types of physiological neuronal activity since physiologically active gap junction currents (GJCs) have never been characterized from single respiratory-related neurons or from single neurons of any other types. In the present study we hypothesized that GJCs could be selectively revealed from single neurons by elimination of transmembrane electrochemical gradients in voltage patch-clamp recording, and this hypothesis was tested in single inspiratory tracheal preganglionic vagal motor neurons (I-TPVMs). The results showed that GJCs were selectively revealed in all I-TPVMs when the transmembrane electrochemical gradients were eliminated in voltage patch-clamp recording, and were rhythmically activated by central inspiratory activity. Therefore, this method may be used as a fast way to detect GJCs within spontaneously active neuronal networks.
Gap Junctions ; physiology ; Motor Neurons ; physiology ; Patch-Clamp Techniques ; Trachea ; cytology

OBJECTIVESTo retrospectively investigate the outcome of transpedicular intracorporeal grafting and posterolateral grafting in treatment of thoracolumbar burst fractures.

METHODSForty-six patients treated with transpedicular intracorporeal grafting from January 1999 to December 2009 and followed up for 19-119 months (average 67 ± 13 months) were reviewed retrospectively, and were compared with 18 patients who had underwent posterolateral fusion during the same period through radiographic analysis. Radiographic measurements included Cobb angle, vertebral wedge angle (VWA), ratio between anterior and posterior vertebral height (APHR), upper inter-vertebral angle, lower inter-vertebral angle on X-ray, CT and MRI.

RESULTSIn transpedicular intracorporeal grafting group, the VWA was corrected from 27.2° ± 6.5° to 7.0° ± 3.0° and the APHR from (53.3 ± 11.8)% to (92.3 ± 2.4)%. In posterolateral fusion group, the VWA was corrected from 23.9° ± 4.4° to 8.8° ± 2.1° and the APHR from (60.7 ± 10.0)% to (88.5 ± 3.3)%. Transpedicular intracorporeal grafting group showed better postoperative correction results than posterolateral fusion group (P < 0.05), and had less loss of correction of Cobb angle, VWA and APHR at final follow-up (P < 0.05).

CONCLUSIONSThe transpedicular intracorporeal grafting can improve injured vertebral body morphology recovery better than posterolateral bone grafting, but can not prevent the late loss of correction after implant removal.

Adolescent ; Adult ; Bone Transplantation ; methods ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVEThe reporting of patient-reported outcomes (PRO) instrument development is vital for both researchers and clinicians to determine its validity, thus, we propose the Preferred Reporting Items for PRO Instrument Development (PRIPROID) to improve the quality of reports.

METHODSAbiding by the guidance published by the Enhancing the QUAlity and Transparency Of health Research (EQUATOR) Network, we had performed 6 steps for items development: identified the need for a guideline, performed a literature review, obtained funding for the guideline initiative, identified participants, conducted a Delphi exercise and generated a list of PRIPROID items for consideration at the face-to-face meeting.

RESULTSTwenty three items subheadings under 7 topics were included: title and structured abstract, rationale, objectives, intention, eligibility criteria, conceptual framework, items generation, response options, scoring, times, administrative modes, burden assessment, properties assessment, statistical methods, participants, main results, and additional analysis, summary of evidence, limitations, clinical attentions, and conclusions, item pools or final form, and funding.

CONCLUSIONSThe PRIPROID contains many elements of the PRO research, and this assists researchers to report their results more accurately and to a certain degree use this instrument to evaluate the quality of the research methods.

Humans ; Outcome Assessment (Health Care) ; Practice Guidelines as Topic ; Research Report ; Research Support as Topic ; Treatment Outcome