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

Female ; Humans ; Kidney Neoplasms ; complications ; pathology ; Ovarian Neoplasms ; pathology ; Wilms Tumor ; complications ; pathology ; Young Adult

Clinical laboratory biological safety is one of whole society safety. This paper introduced briefly the current situation of clinical medical laboratory biosafty in the hospital. and set forth common biological hazards specifically for whose characteristics. Combining the biosafety administration measures from abroad, the issue of laboratory biological safety administration was considered, and put forward some suggestions according to related law and regulation of national laboratory safety administration in order to strengthen clinical laboratory biosafety administration.

A temperature-sensitive mutant strain was isolated after transposon mutagenesis with Tn5 and named MT54.PCR was carried out with primers designed according to the sequence of transposon,the PCR products showed that the MT54 carried transposon in the genome. The mutant grew well at 30℃in minimal medium(MM)containing p-nitrophenol(PNP)as sole carbon source,while it cannot grow at 37℃in the same medium,NO_2.detection results also proved that.Comparing the degradation rate of PNP and hydroquinone of MT54 and DLL-E4 at different temperature,it was speculated that the mutant site locate in the PNP degradation related genes.

Adult ; Altitude ; Craniotomy ; methods ; Follow-Up Studies ; Humans ; Male ; Young Adult

AIM To optimize the subcritical aqueous extraction for polysaccharides from the leaves of Punica granatum L.and to evaluate the in vitro antioxidant activity.METHODS With reaction pressure,solid-liquid ratio,extraction time and extraction temperature as influencing factors,yield of polysaccharides as an evalution index,the extraction was optimized by Box-Behnken method on the basis of single factor test.Then the scavenging effects of polysaccharides on hydroxyl free radical,superoxide anion and DPPH free radical were detected.RESULTS The optimal conditions were determined to be 5 MPa for reaction pressure,1 ∶ 27 for solid-liquid ratio,11 min for extraction time,and 155 ℃ for extraction temperature,the yield of polysaccharides was 1.809％.There was a dose-effect relationship between scavenging rate and polysaccharides' concentration.0.1 mg/mL Polysaccharides displayed the strongest scavenging effects on hydroxyl free radical,superoxide anion and DPPH free radical with the clearance rates of 57.36％,70.51％ and 58.02％,respectively.CONCLUSION This stable and reliable method can be used for the subcritical aqueous extraction for polysaccharides from P.granatum leaves with obvious in vitro antioxidant activity.

Objective To investigate the distribution and antimicrobial resistance of pathogens causing bacterial peritonitis,provide laboratorial guidance for rational use of antimicrobial agents.Methods Pathogenic strains iso-lated from peritoneal fluid specimen of patients with peritonitis in the Affiliated Hospital of Xuzhou Medical Univer-sity in 2011-2015 were collected,performed bacterial identification and antimicrobial susceptibility testing,distri-bution of pathogens and antimicrobial resistance were analyzed.Results A total of 491 strains were collected,in-cluding 291(59.26%)strains of gram-negative bacilli,196(39.92%)of gram-positive cocci,and 4 (0.82%)of fun-gi.The top 5 pathogens were Escherichia coli (30.14%),coagulase negative staphylococcus(12.22%),Staphylo-coccus aureus (10.39%),Klebsiella pneumoniae (8.55%),and Enterococcus faecium(6.52%).Antimicrobial re-sistance rates of Escherichia coli ,Klebsiella pneumoniae ,Acinetobacter baumannii ,and Pseudomonas aeruginosa to imipenem were 4.90%,31.04%,77.28% and 26.27% respectively.Methicillin-resistant Staphylococcus aureus (MRSA)and methicillin-resistant coagulase negative staphylococcus(MRNCS)accounted for 56.02% and 70.02%respectively.Conclusion The main pathogens causing bacterial peritonitis are gram-negative bacilli,Escherichia co-li ranks first;resistance of pathogens is serious,standard use of antimicrobial agents should be strengthened to re-duce the emergence of drug-resistant strains.

Objective To investigate the clinical efficacy of finger reconstruction in child with second toe transplantation,and evaluate the postoperative appearance and function regarding the reconstructed donor feet.Methods From June 2002 to May 2011,sixteen cases were reconstructed in sub-emergency with second toe transplantation.Two thumbs,eight index fingers,and 6 middle fingers were reconstructed.All patients were followed-up from 12 to 24 months.The functions of reconstructed fingers were analysed.Results All the reconstructed fingers survived.Vascular crisis occurred in 1 patient,and survived after re-anastomosis.Necrosis of skin grafts at the domon site with exposed tedons was seen in 1 ease,and healed after changing dressings.All the reconstructed fingers showed good in growth and development,and performed good functions as grabbing,grasping and nipping.Two-point discrimination was between 6 mm and 10 mm.The donor site of the foot had normal gait,without obvious influence on walking.Also,no pain was complained.Conclusion The method of transplanting the second toe can reconstruct the appearance and function of the finger defects in child,and has little effect on the appearance and motion of feet.It is an effective treatment method.

Incorrect drug administration, lack of sexual stimulation, the way of obtaining sildenafil and associating risk factors of erectile dysfunction like severe cardiovascular or neurogenic diseases were the common causes of initial sidenafil failure. To improve the therapeutic efficacy for patients with initial sidenafil failure, education about how to use sildenafil properly plays an important role. Moreover, dose titration, more attempts, a good follow-up and sildenafil taking on a daily basis will yield a high response rate to sildenafil rechallenge.
Adult ; Aged ; Erectile Dysfunction ; drug therapy ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Patient Education as Topic ; Phosphodiesterase Inhibitors ; therapeutic use ; Piperazines ; therapeutic use ; Purines ; therapeutic use ; Sildenafil Citrate ; Sulfones ; therapeutic use ; Treatment Outcome

OBJECTIVETo obtain a simple and effective method to isolate and purify adult Leydig cells.

METHODSThe testes of human adults were digested and then the density gradient centrifugation of the cells was performed with four different Percoll concentrations (60%, 34%, 26%, 21%) to isolate Leydig cells, whose characteristics were identified by cytological observation staining, 3beta-HSD staining and detection of hCG and testosterone secretion.

RESULTSHigh-concentration (> 90%) purified Leydig cells were acquired, and many identification experiments demonstrated the adequate testosterone secretory function of the isolated and purified Leydig cells.

CONCLUSIONThis method is easy and efficient for the isolation and purification of adult Leydig cells.

Adult ; Cell Separation ; methods ; Cells, Cultured ; Chorionic Gonadotropin ; pharmacology ; Humans ; Leydig Cells ; cytology ; drug effects ; secretion ; Male ; Povidone ; Silicon Dioxide ; Testosterone ; secretion