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

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.

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.

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

The biological characters of Sindbis virus strain of Yunnan(YN87448 strain) were studied by the test of the filtration, acid-resistant, ether-resistant, CPE, susceptibility of animal, HA, plague, determination of virus titres, and the cross-HI, cross-IFAT and PRNT as well.The results indicated that YN87448 strain belongs to Sindbis virus, Alphavirus, Togaviridae. YN87448 strain virus was plaque purified(PYN87448). The biological character of PYN87448 strain virus was studied too. PYN87448 strain virus will be used in the molecule biological test.

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.

OBJECTIVETo explore CT and MRI manifestations of the axial area peripheral primitive neuroectodermal tumors (pPNETs) in order to improve the knowledge of this disease.

METHODSThe clinical data of 10 patients with pPNETs underwent pathologically confirmed were retrospectively analyzed from October 2008 to May 2014. There were 7 males and 3 females, aged from 8 to 49 years old with median of 23.6 years. The preoperative multi-slice spiral CT scan was completed in 3 cases, plain CT scan and enhancement in 4 cases; MRI and enhancement scanning in 5 cases; and among them, 2 cases underwent both MRI and CT scan.

RESULTSIn-bone type was found 6 cases and out-bone type was found 4 cases. Three cases occurred in sacral vertebrae, 2 cases in lumbar vertebrae, 1 case in cervical vertebrae, 1 case in cervical spinal canal, 1 case in coccyx, 1 case in the right iliac bone, 1 case in presacral space. Cross sectional the smallest tumor maximum level was 1.1 cmx 1.2 cm in size, the biggest tumor was 8.0 cm x 9.2 cm, the median size was 4.4 cm x 5.7 cm, of them, the tumor of maximal diameter larger than 5 cm had 6 cases. Except 2 cases-without destruction of bone, the other 5 cases with osteolytic destruction, 2 cases with calcification, 1 case with mixed. Equidensite was main in CT scan, 1 case with uniform density, other 6 cases with uneven density,in which 3 cases with "floating ice" change; 1 case with moderate strengthening, other 3 cases with obviously strengthening, 2 cases with multiple small blood vessels in enhancement scanning. MRI of 5 cases showed the signal of isointensity on T1WI, the slightly high signal on T2WI and the signal was not uniform; after enhancement scan, the signal of 5 cases obviously enhanced. Two patients complicated with vertebral compression fractures, no periosteal reaction was found in all patients, and no the destruction of intervertebral disk was found in 5 patients of MRI scan.

CONCLUSIONThe axial area pPNETs is common among children and the youth, and the mass often is huge. The mass of in-bone type often envelopes the vertebral body, and main located on prevertebral space, all associated with bone destruction, osteolytic destruction is common, and primary vertebral bodies also is common, attachment primary or involvement is few found, it can involve the spinal canal and anterior wall of spinal canal is common, some cases complicate with multiple newly born small vessels. The mass of out-hone type in deep soft tissue is common, minority primary spinal canal, many complicated with vertebral bone destruction, osteolytic destruction was main. The intervertebral disk was not invaded and intervertebral space has not stenosis. CT scan offer complicate with "floating ice" sign, and in-bone type is common. Isointensity is main on MRI TlWI and slightly longer signal is main on MRI T2WI, strengthening signal is obvious.

Adolescent ; Adult ; Child ; Cross-Sectional Studies ; Diagnosis, Differential ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neuroectodermal Tumors, Primitive, Peripheral ; diagnosis ; diagnostic imaging ; Radionuclide Imaging ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo investigate the effect of transurethral resection of ejaculatory ducts (TURED) for azoospermia with ejaculatory duct obstruction (EDO).

METHODSFrom June 2003 to December 2004, 20 azoospermia with EDO were diagnosed, diagnostic criteria included a history, physical examination, semen analyses, semen fructose measurement, endocrine assessment, testicular biopsy and transrectal ultrasonography (TRUS); All 20 cases were treated by TURED. Fifteen of them were followed up more than 3 months after the treatment. The semen samples of them were analysed at 3-month intervals in post-therapy.

RESULTSSemen analyses in all 20 cases showed the typical characteristics of EDO, low semen volume (0.4-1.6 ml), azoospermia, low pH, absent or low semen fructose. TRUS showed the main etiology factor of EDO was a midline cyst in 11, lateral cystic lesions in 2, the remaining 7 cases had dilated ejaculatory duct with or without dilated seminal vesicles. Among 15 cases followed up more than 3 months after TURED, 10/15 (67%) had an improvement in semen parameters and 3/15 (20%) had pregnancies. Semen analyses had not been done in anther 5 cases.

CONCLUSIONTransurethral resection of ejaculatory ducts may be a safe and effective method for the treatment of azoospermia with EDO.

Adult ; Azoospermia ; diagnosis ; surgery ; Ejaculatory Ducts ; diagnostic imaging ; pathology ; surgery ; Electrosurgery ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Oligospermia ; diagnosis ; Ultrasonography

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