We have obtain a steady and reliable dyeing methods for the uniuncleate and dicaryotic hyphae of Pleurotus tuber-regium by using different foster hyphae way, comparing two kinds of fastness liquid and three dye stuff on the hyphae nuclear stain effect, and then optimization grouping.

OBJECTIVETo investigate the triterpenoids chemical constituents in rhizome of Alisma gramineum.

METHODSilicon gel chromatography and HPLC techniques were employed for isolation and purification of the constituents, and the structures were elucidated by various spectral analyses such as NMR, MS and IR.

RESULT5 protostane-type tetracyclic triterpenoids were isolated and identified as 16,23-oxidoalisol B(1), 11-deoxyalisol C(2), alisol F(3), 16beta-methoxyalisol B acetate (4), 16beta-hydroxyalisol B acetate (5).

CONCLUSIONAll of these compounds were found from A. gramineum for the first time.

Alisma ; chemistry ; Cholestanones ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Triterpenes ; chemistry ; isolation & purification

Objective To repair segmental mandibular defects with autogenous bone marrow stromal cells(BMSCs)and?-triealcium phosphate.Methods Isolated BMSCs were in vitro expand- ed.A 3 cm-long segmental mandibular defect was created at right mandible in 12 canines,of which de- fects in six canines were repaired with BMSCs and?-tricalcium phosphate(?-TCP)and that in other six cases repaired with?-TCP,which was used as control.The engineered bone was evaluated by X-ray, CT,DXA,gross and histological examination,immunohistochemistry and biomechanical test 4,12,26,32 weeks after operation respectively.Results In induced BMSCs,histochemistry showed AKP activity. Oral X-ray showed obvious callus formation 4-26 weeks after operation in experimental group but minimal bone formation in control group.At 32 weeks after operation,gross observation,X-ray and CT demonstra- ted well bony-union in experimental group but bony-nonunion in control group.DXA indicated that the bone density of experimental group was significantly higher than that of control group.Biomechanical test revealed no statistical difference upon mechanical strength of mandibula between experimental group and normal group.Conclusions Canine segmental mandibular defects can be well repaired with the tissue- engineered bone generated by autogenous osteogenic BMSCs and?-TCP scaffold.

AIMTo study the protective effect of sodium pyruvate on ischemia/reperfusion injury following hemorrhagic shock.

METHODSRat models of hemorrhagic shock were built up. When the shed blood was infused, the rats were also randomly provided by one of normal saline, glutathione and sodium pyruvate. Rats were killed 3 hours after the reperfusion, the activity of plasma lactate dehydrogenase (LDH) and glutamic-oxaloacetic transaminase (GOT), the level of tissue malondialdehyde (MDA) and the activity of tissue myeloperoxidase (MPO) were detected. Biopsy specimens were obtained to investigate morphological changes of the myocardial, hepatic, lung and renal tissue.

RESULTSThe activity of plasma LDH and GOT, the level of MDA of hepatic, lung and renal tissue and the activity of MPO of myocardial, lung and renal tissue decreased remarkably in group given sodium pyruvate compared with group given normal saline, and the effect of group given sodium pyruvate was more remarkable than group given glutathione.

CONCLUSIONThese data support the view that sodium pyruvate shows protective effect on ischemia/reperfusion injury following hemorrhagic shock. It is possibly relevant to scavenging of oxygen free radicals, reduction of neutrophil, and anti-inflammatory response.

Animals ; Aspartate Aminotransferases ; blood ; Disease Models, Animal ; Kidney ; metabolism ; pathology ; L-Lactate Dehydrogenase ; blood ; Liver ; metabolism ; pathology ; Lung ; metabolism ; pathology ; Male ; Malondialdehyde ; analysis ; Peroxidase ; analysis ; Protective Agents ; pharmacology ; Pyruvic Acid ; pharmacology ; Rats ; Rats, Wistar ; Reperfusion Injury ; blood ; pathology ; prevention & control ; Shock, Hemorrhagic ; blood ; pathology

OBJECTIVETo compare the therapeutic effect of recombinant human brain natriuretic peptide (rhBNP) and nitroglycerin on acute decompensated heart failure (ADHF).

METHODSFifty ADHF patients were randomly divided into rhBNP group and nitroglycerin group. In all the patients, dyspnea and global clinical status were assessed before and at 30 min, 6 h and 24 h after drug administration, and the volume of fluid intake and urine along with hemodynamic parameters was recorded 24 h after drug administration. In the nitroglycerin group, the patients received an initial nitroglycerin dose of 5 microg/min, with subsequent dose increment of 5 microg/min every 3 to 5 min; the dose was adjusted individually according to the hemodynamics of the patients. The patients in rhBNP group were given rhBNP at the initial dose of 1.5 microg/kg by with an intravenous bolus injection followed by infusion at the rate of 0.0075 microg.kg(-1).min(-1) for 72 h.

RESULTSAt 30 min and 6 h after drug administration, the patients in the rhBNP group showed significant greater improvement of dyspnea (P=0.042 and 0.019) and global clinical status (P=0.018 and 0.044) than those in the nitroglycerin group, but 24 h after drug administration, no significant difference was noted between the two groups (P=0.192 and 0.179). Twenty-four hours after drug administration, the mean urine volume was significantly greater in rhBNP group than in nitroglycerin group (1513.8-/+242.9 vs 1341.2-/+239.7 ml, P=0.015), and the ejection fraction increased and pulmonary arterial pressure and systolic blood pressure decreased at greater amplitude in the former group (P=0.001,0.000 and 0.002, respectively). At 72 h, the numbers of premature ventricular contraction and couplets premature beats and onset of paroxysmal ventricular tachycardia were significantly reduced in rhBNP group as compared with the nitroglycerin group (P=0, 0.001 and 0.002, respectively).

CONCLUSIONRhBNP promotes urine excretion, decreases pulmonary arterial pressure and increases left ventricular ejection fraction to improve dyspnea and global clinical status and reduce the onset of ventricular arrhythmia in ADHF patients.

Aged ; Aged, 80 and over ; Blood Pressure ; drug effects ; Female ; Heart Failure ; drug therapy ; pathology ; physiopathology ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; administration & dosage ; genetics ; therapeutic use ; Nitric Oxide Donors ; administration & dosage ; therapeutic use ; Nitroglycerin ; administration & dosage ; therapeutic use ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Treatment Outcome

Cadherins ; metabolism ; Female ; Humans ; MCF-7 Cells ; Neoplastic Stem Cells ; metabolism ; Receptor, Notch1 ; metabolism ; Receptors, CXCR4 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism

OBJECTIVETo provide data on alcohol consumption and drug use among middle-school students aged 13-15 in 4 cities of China, and to provide evidence for developing intervention strategies on adolescents alcohol and drug use.

METHODSStandardized sample selection process of two-stage cluster-sampling was used in middle-school students in Beijing, Hangzhou, Wuhan and Urumchi. A self-administered questionnaire survey was conducted in Sept. 2003 and data was analyzed by Epi Info software.

RESULTSAmong 7344 students from grade 1 to 3, 36.5% had tasted while 14.4% had drunk alcohol in the past 30 days. 9.9% had experienced drunkness, 5.1% had been in trouble because of drinking, and 1.6% had ever used illegal drugs. Significant differences had been found in all the cities. Higher graders, older students and boys had higher rates of alcohol and addictive drug use than low graders, younger students and girls. 51.9% had been taught on take alcohol safety and another 27.6% on skills of rejecting alcohol, during the past school year.

CONCLUSIONSThe current situation of alcohol and addictive drug use among Chinese middle-school students aged 13-15 seemed to be quite critical, suggesting that it is necessary to carry out relevant health education in accordance with different characteristics in area, gender and age of the students.

Adolescent ; Alcohol Drinking ; epidemiology ; China ; epidemiology ; Female ; Humans ; Male ; Students ; Substance-Related Disorders ; epidemiology

To investigate the effect of GFP fused to C terminal of integrin alpha(IIb) on the biosynthesis and expression of alpha(IIb) beta(3) compound, the alpha(IIb) GFP expression plamid, named palpha(IIb) GFP, the cDNA of alpha(IIb) was constructed from p3.1-2b and fused to pEGFP-N1 in frame. When the sequence of palpha(IIb) GFP was confirmed by sequencing it was transferred to Chinese Hamster Ovary (CHO) cells with or without p3.1-3a expressing integrin beta(3). Then the expression of alpha(IIb) GFP fusion protein was confirmed by Western blot and then its subcellular localization was determined with laser confocal scanning microscopy. The results showed that the target gene was cloned into recombinant vector by restriction analysis and sequencing. Overexpression of the fusion protein in the transfected CHO cells was identified with Western blot. Subcellular localization analysis confirmed that alpha(IIb) GFP was expressed in CHO cells and could be transferred from endoplasmic reticulum to Golgi apparatus. It is concluded that the eukaryotic expression plasmid containing alpha(IIb) GFP fusion gene is successfully constructed. GFP fused to the cytoplasmic tail of integrin alpha(IIb) allows the normal expression of alpha(IIb) beta(3) in CHO cells.
Animals ; Blotting, Western ; CHO Cells ; Cricetinae ; Cricetulus ; Endoplasmic Reticulum ; metabolism ; Golgi Apparatus ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Microscopy, Confocal ; Platelet Glycoprotein GPIIb-IIIa Complex ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Chitosan microsphere has been wildly researched in controlled release of protein and peptide drug because of its excellent mucoadhesive and permeation enhancing effect across the biological surfaces. The control of the size and size distribution of microspheres is necessary in order to improve reproducibility, bioavailability, and repeatable release behavior. In this work, uniform-sized chitosan microspheres containing insulin were prepared by a novel membrane emulsification technique combined with glutaraldehyde crosslinking method. In order to prepare uniform-sized chitosn microspheres, it is necessary to modify hydrophilic membrane into hydrophobicity. It is found that there exists a linear relationship between the size of chitosan microspheres and pore size of the membrane used, so it is easy to control the size of microspheres by using membranes with different pore size. In this study, the effect of different amount of crosslinker and crosslinking time on microspheres' morphology, encapsulation efficiency (EE) and release profile of drug in vitro were investigated. It is shown that the morphology of microspheres is more smooth and spherical, and the release rate is slower with the increase of amount of glutaraldehyde and prolongation of crosslinking time. When the molar ratio of amino group of chitosan to aldehyde group of glutaraldehyde is 1:0.7, and crosslinking time is 1 h, the highest EE was obtained (about 65%). Date obtained suggest that chitosan microspheres prepared by this new method would be a promising system for controlled release of protein drugs.
Biocompatible Materials ; chemistry ; Chitosan ; chemistry ; Cross-Linking Reagents ; Delayed-Action Preparations ; chemical synthesis ; Drug Carriers ; chemical synthesis ; Emulsions ; Glutaral ; chemistry ; Humans ; Insulin ; pharmacokinetics ; Microspheres ; Particle Size

OBJECTIVETo explore the anti-tumor mechanism of the combination of cisplatin with DC vaccine in tumor-bearing mice.

METHODSB16 melanoma cells were treated with cisplatin at the final concentration of 20 µg/ml in vitro for 24 h. The expression of HMGB1, Hsp70 and TGF-β were detected by Western blot. B16 tumor-bearing mouse models were generated. The therapeutic effect of the combination of cisplatin (100 µg/mouse i.p., for sequential 3 days) and intratumoral injection of DC cells (3×10(6)/mouse, twice with a 7-day interval) in the tumor-bearing mouse models was evaluated. Expression of MHC II, ICAM-1 and CD86 was analyzed by flow cytometry. The mice were sacrificed at 28 days after tumor cell inoculation. The tumors were removed and weighed, and tissue samples were taken for pathological examination. Tumor infiltrating lymphocytes (TIL) were isolated by discontinuous gradient centrifugation. The distribution of T-reg and CD8(+) T cells in the TIL was analyzed by flow cytometry, and the ratio of CD8(+) T/T-reg was determined. The activity of cytotoxic lymphocytes (CTL) was determined by microcytotoxicity assay.

RESULTSCisplatin enhanced both the B16 cell apoptosis and HMGB1 expression. After loading with cisplatin-treated cell lysate, the expression of MHC II, ICAM-1 and CD86 on DC cells were (47.5 ± 8.8)%, (35.5 ± 8.3)% and (36.2 ± 9.2)%, respectively. At 28 days after tumor cell inoculation, the tumor weight of the control group was (2.1 ± 0.6) g, that of the cisplatin group was (0.3 ± 0.2) g and that of cisplatin + DC vaccine group was (0.5 ± 0.2) g, showing a significant inhibition of tumor growth (P < 0.01). Furthermore, the CD8(+) T/T-reg ratio and CTL activity in TIL were also significantly enhanced in the tumor-bearing mice treated with cisplatin + DC vaccine. When the effector-to-target ratio was 20:1, 10:1 and 5:1, the CTL activity in the cisplatin + DC vaccine treated mice was (25.0 ± 5.0)%, (22.0 ± 6.0)% and (14.0 ± 4.0)%, respectively, significantly higher than (8.2 ± 3.6)%, (6.7 ± 1.8)% and (3.6 ± 1.9)%, respectively, in the control group (all P < 0.01).

CONCLUSIONCisplatin promotes the anti-tumor effect of DC vaccine by down-regulating T-reg cells and enhancing the CTL activity in tumors.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; B7-2 Antigen ; metabolism ; CD8-Positive T-Lymphocytes ; pathology ; Cancer Vaccines ; pharmacology ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Dendritic Cells ; immunology ; metabolism ; Female ; Genes, MHC Class II ; HMGB1 Protein ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Melanoma, Experimental ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; T-Lymphocytes, Cytotoxic ; immunology ; T-Lymphocytes, Regulatory ; pathology ; Tumor Burden ; drug effects