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

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

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

OBJECTIVETo construct a phosphatidylinositol 4-kinase beta (PI4K-beta) mutant with the 325th to 373rd amino acid codons deleted, and try to develop a simple method for constructing middle fragment deletion mutant.

METHODSIn line with the mechanism of gene splicing by overlap extension(SOE), an additional PCR was used to get the PI4K-beta mutant in which the 325th to 373rd amino acid codons were deleted. Then the mutated gene was cloned into pCMV-Tag4A mammalian expression vector.

RESULTSA mutant with the 325th to 373rd amino acid codons deleted was constructed successfully.

CONCLUSIONThe improved SOE is a very effective and reliable method to construct middle fragment deletion mutant. It is worthy to be popularized.

1-Phosphatidylinositol 4-Kinase ; genetics ; Base Sequence ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; genetics ; Polymerase Chain Reaction ; methods ; Protein Engineering ; methods ; Recombinant Proteins ; genetics ; Sequence Deletion

The aim of the present study was to investigate the effect of glucagon-like peptide-1 (GLP-1) on palmitate-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and the underlying mechanism. HUVECs were cultured in vitro, and then treated by palmitate to induce apoptosis. Meanwhile, GLP-1 was added to explore its effect. After 24 h of the treatments, Caspase-3 activity and DNA fragmentation were measured using ELISA kits. Phospho-p38 mitogen-activated protein kinase (p-p38 MAPK) expression was detected by Western blot. The results showed that incubating HUVECs with 0.125 mmol/L GLP-1 increased Caspase-3 activity and DNA fragmentation. GLP-1 significantly inhibited palmitate-induced increases of Caspase-3 activity and DNA fragmentation in a concentration-dependent manner. Moreover, GLP-1 inhibited the up-regulation of p-p38 MAPK expression induced by palmitate in HUVECs. These results suggest GLP-1 protects HUVECs against lipo-apoptosis, and this effect may be mediated through inhibiting p38 MAPK pathway.
Apoptosis ; Caspase 3 ; metabolism ; DNA Fragmentation ; Glucagon-Like Peptide 1 ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; MAP Kinase Signaling System ; Up-Regulation ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo research the changes in microscopic characteristics and ability of secreting testosterone between aged SD rat Leydig cells and young SD rat Leydig cells.

METHODSThe total and free serum testosterone levels of serum both young and aged rats were examined. The changes in microscopic characteristics between young and aged rat Leydig cells were observed under microscope and electron microscope. The testosterone secreted by cultured Leydig cells of stimulated by hCG and Forskolin both groups were detected.

RESULTSA significant difference was found in both total and free testosterone levels between young and old rats (P < 0.05). Aged SD rat Leydig cells were observed smaller in volume and more markedly stained than young ones; The secretion ability of aged rat Leydig cells was found lower than that of young rat Leydig cells with or without hCG and Forskolin stimulation (P < 0.05).

CONCLUSIONThe secreting index of aged SD rat Leydig cells is lower than that of young rat Leydig cells both in vivo and vitro, and the reason is the system of synthesizing testosterone is arrested.

Androgens ; deficiency ; Animals ; Cells, Cultured ; Leydig Cells ; pathology ; secretion ; Male ; Progesterone ; secretion ; Rats ; Rats, Sprague-Dawley ; Testosterone ; secretion

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