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

Objective To evaluate the appropriate timing of coronary artery bypass grafting(CABG) with acute myocardial in- farction(AMI) and to discuss the influence of postoperative mortality on 30 days.Methods 233 patients after CABG were divided into 2 groups,AM/group and unstable angina (UA) group.There were 176 males (75.4%) and 57 females (24.5%).The mean age was (65.6?9.2) years(range 34～86 years).The mean grafts were 3.46?0.89.The complex risk elements between the 2 groups were analyzed to evaluate the independent risk element of death.Results Internal mammary arteries were used in 137 patients (58.8%).The postoperative mortality rate was 4.3 % (10/233).The operative mortality rates(OMR) were closely related to the in- creasing time intervals between AMI and CABG,for less than 3 days was 14.6% (6/41 cases),for 4 to 10 days was 2.7 % (1/37) and for 11 to 30 days was 0.The OMR of AMI less than 3 days has significant difference (P=0.033) comparing with that of unstable an- gina pectoris [2.3% (3/130) ].Conclusion Proper timing of CABG should be done in 3 days after AMI.

Objective To investigate the relationship between the expression of decay receptor 3 (DcR3) and the eliniealpathological parameters in human gastric carcinoma. Methods The expression of DcR3 was examined by RT-PCR in a series of 41 human primary gastric carcinomas and 41cases of normal tissue adjacent to tumor. Multiple clinical pathological factors were analyzed according to their relation with the expression of DcR3. Results The positive rate of expression of DcR3 was 56 %(23/41) in human gastric carcinoma. The expression of DcR3 in gastric carcinoma was significantly higher than that in normal tissues adjacent tumor. The expression of DcR3 was significantly correlated with different degrees of differentiation, lymph node metastasis and TNM staging (P <0.10), but there was no significant difference in DcR3 and other clinical pathological features such as tumor position and invasion depth (P>0.10). The multiple linear regression equation was Y=0.432-0.208X1+0.098X2+0,086X3. Conclusion DcR3 expression can be highly found in gastric carcinoma. The abnormal expression of DcR3 may promote tumorigenesis and progression. DcR3 may be important in evaluating the tumor differentiation, infiltration depth, lymph node metastasis and TNM staging of human gastric carcinoma.

Animals ; Athletic Performance ; Drugs, Chinese Herbal ; pharmacology ; Free Radicals ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Morinda ; Myocardium ; metabolism ; Physical Conditioning, Animal

AIMTo prepare insulin powder for inhalation by spray-drying technology, determine the deposition of the insulin powder formulation in vitro and preliminarily investigate hypoglycemic response of the dry powder with/without absorption promoters.

METHODSThe depositions of the insulin powder for inhalation were determined by the China Pharmacopoeia 2000 version addenda XH and hypoglycemic effects were evaluated by testing serum glucose with glucose oxidase-peroxidase (GOD-PAP) method.

RESULTSThe depositions of the spray-dried insulin powder for inhalation were more than 40% under various humidity and their changes were not significant when air flow was no less than 18 L x min(-1). The coadministration of insulin with 8 mmol x L(-1)/dose sodium taurocholate [PA = 59.91%, Cnadir = (33 +/- 6) %] and 10 mmol x L(-1)/dose sodium deoxycholate [PA = 47.46% , Cnadir = (32 +/- 7)%] induced a significantly greater decline in blood glucose levels, while coadministration with 1% sodium caprylate, 1% sodium dodecyl sulfate, 250 microg/dose lecithin, 10 mmol x L(-1)/dose EDTA appeared to have no significant effect (P > 0.05).

CONCLUSIONInsulin powder for inhalation was relatively stable under various humidity conditions and different flow current. The use of 8 mmol x L(-1)/dose sodium taurocholate and 10 mmol x L(-1)/dose sodium deoxycholate could be able to potentially improve the bioavailability of insulin by pulmonary route.

Administration, Inhalation ; Animals ; Biological Availability ; Blood Glucose ; metabolism ; Deoxycholic Acid ; pharmacology ; Drug Synergism ; Female ; Humidity ; Hypoglycemic Agents ; administration & dosage ; pharmacology ; Inhalation ; Insulin ; administration & dosage ; pharmacology ; Male ; Powders ; Rats ; Rats, Sprague-Dawley ; Taurocholic Acid ; pharmacology

Objective To explore the role of NF-kB activation on spontaneous formation of germinal centers in spleen in BXSB mice and it's mechanisms.Methods Eighteen BXSB mice were divided to control group and pyrrolidine dithiocarbonate(PDTC)group randomly.PDTC group was given PDTC 120 mg/kg?BW ip every other day and control group was given the same dose of dissolving solution.NF-kB activity was deter- mined by electrophoretic mobility shift assay.Two color flow cytometry were used to detect CD154 expression on splenic B cells and germinal center B cells apoptosis.Germinal centers were stained for histochemical analysis.Results PDTC could inhibit the NF-kB activity in spleen tissue in BXSB mice.It decreased the NF-kB activity by 62.82%.Spontaneous germinal center formation was detected in spleen in BXSB mice.In- hibiting NF-KB activation could down-regulate CD154 expression on splenic B cell,retard spontaneous germi- nal center formation and increase germinal center B cell apoptosis.Conclusion NF-kB activation may induce spontaneous germinal center formation in spleen in BXSB mice by upregulating CD154 expression on splenic B cell and decreasing germinal center B cell apoptosis.The autoreactive B cells generated during spontaneous germinal center formation may escape apoptosis and then differentiate to autoantibody-producing plasm cells.It suggests that NF-kB can be a therapeutic target.

Objective To explore humam cytomegalovirus(HCMV) encoded microRNAs during latent infection in order to help study HCMV virology and latent infection mechanisms.Methods A model of HCMV latent infection via THP-1 cells infected with HCMV was constructed.Deep-sequencing was performed using high-resolution Solexa sequencing platform.The secondary structure of the newly sequenced miRNA was predicted by RNAFold bioinformatics software. Results HCMV encoded various miRNAs during latent infection, including miR-US25-2-3p, miR-US25-2-5p, miR-UL112, miR-US25-1, miR-UL22A and PC-5p-148467 with a predicted length of 25 bp, named hcmv-miR-US33as-5p.Conclusion HCMV can express many types of miRNAs during latent infection.

Objective:To study the differentiation capacity of the fibroblast-like cells isolated from human skin dermis into mesenchymal stem cells, and to explore the feasibility to use these cells as alternative cell source of autologus bone marrow mesenchymal stem cells (BMSCs ) for regeneration of tissue inj uries and defects. Methods:Full thickness skin samples were obtained from the abdomen of surgical patients, then digested with dispase and collagenase Ⅰ subsequently. Thereafter, the digested cells were collected and cultured, followed by suspension with serum free medium containing N2,B27,basic fibroblast growth factor (bFGF),and epidermal growth factor (EGF).The skin dermis derived spheroids (SDDSs)were collected and monolayer cultured in serum-containing medium.Finally,the cells were characterized by immunofluorescence staining and differentiation assays.Results:The dermis derived cells proliferated and formed SDDSs in the suspension of serum-free medium. After monolayer cultivation in serum-containing medium, the cells from spheroids were successfully expanded to large number. The cells expressed mesenchymal stem cells markers CD90, CD105 and vimentin. Under osteogenic,chondrogenic and adipogenic differentiation conditions,these cells were differentiated into the alizarin red,safranin O, and oil red O staining positive cells, displayed similar differentiation traits with BMSCs. However,safranin O staining was weaker in the dermis derived cells than BMSCs. Conclusion:A kind of fibroblast-like cells exist in human skin dermis, and have osteocytic, chondrogenic and adipogenic differentiation potentials,demonstrating that these cells will be utilized as a novel cell source for repairing the tissue injury and defect in clinic.

OBJECTIVETo investigate the effect and mechanism of Compound Salvia injection (CSI) on nitrate ester tolerance.

METHODSEighty-four patients with coronary heart disease (CHD) were randomly divided into three groups, Group A treated with isosorbide dinitrate (ISD, 15 mg, 4 times per day) alone, Group B with ISD plus CSI and Group C with ISD plus vitamin C. The therapeutic course for all groups was 10 days. The tolerance to nitrate ester and blood pressure were monitored. Before and after treatment, the color Doppler ultrasonic apparatus was used to detect the baseline value of humeral arterial internal diameters (D0), the humeral arterial dilatory response under compression [D1, that is, the flow-mediated vasodilation (FMD)] and the vasodilatory response after sucking of nitroglycerin (D2). And the blood levels of endothelin-1 (ET-1), endothelial nitric oxide synthase (eNOS) mRNA expression were determined. The endothelial-dependent vasodilation (EDD) was expressed by (D1 - D0)/D0 x 100%, and the endothelial-independent vasodilation (EID) was expressed by (D2 - D0)/D0 x 100%.

RESULTS(1) The occurrence rate of nitrate tolerance in Group B and C (28.57% and 35.7%) was lower than that in Group A (64.29%), but insignificant difference was found between the former two. (2) After treatment, blood pressure increased in Group A to the level of pre-treatment, that in Group C also increased but still lower than that of pre-treatment, while insignificant increase was observed in Group B, comparison between Group B and C showed significant difference (P < 0.05). (3) After treatment, EID lowered in Group A, EDD increased in Group B and C (P < 0.05), EDD and EID in Group B and C were higher than those in Group A (P < 0.05), and EDD was higher in Group B than in Group C (P < 0.05). (4) After treatment, ET-1 mRNA expression lowered in Group B, eNOS mRNA expression increased in Group B and C, with significant difference as compared with those before treatment and those in Group A (P < 0.05), and eNOS mRNA expression in Group C was lower than that in Group B (P < 0.05).

CONCLUSIONCSI could partially prevent the occurrence of tolerance to nitrate ester, with the effect better than vitamin C, the mechanism might be related with its regulation on eNOS, ET-1 mRNA expression and protection on vascular endothelial function.

Adult ; Aged ; Coronary Disease ; drug therapy ; Drug Resistance ; Drugs, Chinese Herbal ; administration & dosage ; Endothelin-1 ; biosynthesis ; genetics ; Female ; Humans ; Injections, Intravenous ; Isosorbide Dinitrate ; therapeutic use ; Male ; Middle Aged ; Nitric Oxide Synthase ; biosynthesis ; genetics ; Nitric Oxide Synthase Type III ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Salvia miltiorrhiza ; Vasodilator Agents ; therapeutic use

OBJECTIVEHirschsprung's disease (HD), one of the most common causes resulting in lower intestinal obstruction in children, is prone to be misdiagnosed or to be missed from diagnosis because of its atypical clinical symptoms and inconspicuous morphological findings by barium enema X-ray. Recently, this situation has been largely ameliorated by increased comprehension of anorectal kinetics and improvement of instrument for measurement of anorectal pressure. By now, anorectal manometry (ARMM) has been regarded as a routine means for functional assessment and diagnosis for anorectal disease. Nevertheless, the accuracy rate of diagnosis of HD in neonate by ARMM remains to be elucidated. In this study the clinical evaluation of anorectal manometry as an early diagnostic method for neonates with Hirschsprung's disease was appraised.

METHODSForty-two HD patients defined by pathological study of rectal tissue obtained via rectal mucous membrane biopsy or operation were recruited in this study. ARMM was performed in liquid transmission using PC polygraph high rate gastrointestinal dynamical detection system (PC Polygraf HR, CTD-synectics, Sweden), with 4-lumen catheter with which a small 5-cm-long balloon was connected at the terminus. All children were positioned on their left side or back during the procedure and the pressure transducers were placed in the mid-axillary line level. The results of ARMM performed before operation or biopsy were compared with the results of barium enema X-ray testing. The decrease of internal anal sphincter pressure as rectoanal inhibitory reflex (RAIR) was measured based on the fluctuation curve of pressure detected. HD was defined when no decrease of anal catheter pressure was detected after insufflation (RAIR positive), and suspected HD state was assessed with the presentation of incomplete relaxation or positive/negative results coexisted (RAIR abnormal) in canal.

RESULTSThirty patients (71.43%) were diagnosed as HD by ARMM including 18 patients who showed negative response to RAIR and 12 patients whose response was abnormal. While barium enema examinations were carried out in all the 45 patients, the results showed 5 HD patients and 14 suspected HD patients, giving an overall diagnostic accuracy of 45.24%. There were also 16 patients with positive ARMM response and negative barium enema findings together, and 5 patients with negative ARMM results and positive barium enema findings at the same time. There was a significant difference between the two diagnostic methods (chi(m)(2) = 4.76, P < 0.05).

CONCLUSIONAnorectal manometry seems to be a more reliable method for diagnosis of Hirschsprung's disease in neonate than barium enema X-ray. Because ARMM is a simple, safe and non-invasive method, it can be used as a screening test of choice in neonates with clinically suspected HD. But for final diagnosis, it is reasonable to combine ARMM with other diagnostic methods in HD patients.

Anal Canal ; physiopathology ; Barium Sulfate ; Enema ; Hirschsprung Disease ; diagnosis ; Humans ; Infant, Newborn ; Manometry ; Rectum ; pathology ; physiopathology