The concept of humanistic education is based on the concern of human beings and their abilities of development. Thus,it is an educational concept to optimize the teaching modules.It stresses the point that education must regard the develop- ment of human beings as the main line and human beings should be emphasized in education from the very beginning to the end. Case-Teaching Method which takes the students as the main part fully manifests the concept of humanistic education,in which all teaching activities and the processes closely revolve the students in order to train them to carry on their studies and to solve the problems independently.

Objective Previous studies indicated that activation of Toll-like receptor4 (TLR4) was involved in the progression and instability of atherosclerotic plaque.Anti-inflammatory effects were shown in statins. However,the mechanisms underlying these effects have not been well explored.We test the hypothesis that a por- tion of these anti-inflammatory effects are mediated by regulation of TLR4 expression.Methods One hundred twenty-one subjects (22 normal persons,17 patients with stable angina and 82 patients with ACS) were recruited. 41 patients with ACS were randomized to atorvastatin 10 mg/d or atorvastatin 40 mg/d on top of routine anti-anginal treatment.Serum level of hsCRP,blood lipids,TLR4 expression on CD_(14)~+ monocytes were measuered before and after one month treatment.TLR4 expression on CD_(14)~+ monocytes were quantified via flow-cytometry.Results hsCRP and TLR4 expression on CD_(14)~+ monocytes in patients with ACS were higher than patients with stable angina and normal persons(hsCRP,ACS:11.1?14.3 vs stable angina:2.5?2.7 mg/L vs normal:2.3?4.2 mg/L,P

Objective To establish a specific,sensitive,simple human respiratory adenovirus detection method to put a good experimental foundation for developing universal and specific diagnostic reagents of human respiratory adenovirus.Methods The nucleotide sequences of 10 serotype of human respiratory adenovirus were obtained from GenBank.Highly consewed five pairs of universal and specific adenovirus primers were designed on the evaluation of multiple sequence alignment of the 10 full genomic sequences with the software DNAMAN 5.2.2,Gene Runner 3.05,BLAST,and to ensure that the polymerase chain reaction(PCR)products were type-specific.NP-40 sample lytic method was employed to prepare the template.The effectiveness,specificity and sensitivity of primers were evaluated.And PCR was carried out to test 64 samples of throat swabs of acute respiratory infection children.The positive PCR products were sequenced directly to identify the adenovirus serotypes.The positive specimens was inoculated on HeLa cells to observe cytopathie effect(CPE)under light microscope,and virus morphology were observed under electron microscope.Results BLAST results indicated that the five pairs of primers were specific adenovirus primers with low homology to the others.The primers were identified as PCR positive fragments obtained by using five pairs of primers to amplify the human adenovirus type 3 DNA,which did not react to the respiratory syncytial virus and Coxsackie virus.the effectiveness and specificity of the primer were thus indicated.PCR sensitive results showed 10-5 dilutin of adenovirus culture and DNA sample could be deteeted which meant the method was sensitive and stable.Two PCR positive specimens were detected in 64 clinical samples.the positive rate was 3.13%(2/64).Using the two PCR positive specimens to inoculate the HeLa cells,the typical adenovirus CPEs of rounded and aggregated,detatched cells were observed under light microscope.And a large number of adenovirus typical particles with characteristic lattice arrangement were observed in the infected cells under electron microscope.PCR product sequencing results showed that these two isolated adenoviruses were typed in human adenovims group B.Conclusions Universal and specific adenovirus PCR primers with the serotype specificity of the PCR products were designed successfully.The PCR primers was sensitive and specific and could be routinely applied in clinical adenovirus diagnosis for respiratory specimens.

OBJECTIVE: To determine the expressions of Notch1, Jagged1 and vascular endothelial growth factor (VEGF) in human non-small cell lung cancer (NSCLC) and to explore its clinical pathological significance. METHODS: Immunohistochemical SP method was used to detect the expressions of Notch1, Jagged1 and VEGF in 65 patients with NSCLC and 15 normal epithelial tissues of the lung, and the relationship between them and clinic-pathological parameters were analyzed. RESULTS: The positive rates of Notch1, Jagged1 and VEGF in NSCLC were 81.5%, 83.1% and 93.8%, respectively, higher than those in normal epithelial tissues of the lung (P<0.05). The positive expression levels of Notch1 and VEGF were closely associated with the tumor stage and the lymph node metastasis (P<0.05). The positive expression levels of Jagged1 was positively correlated with the pathological type and lymph node metastasis. There was a positive correlation between Notch1, Jagged1 and VEGF. CONCLUSION Notch1, Jagged1 and VEGF protein may play an important role in the pathway of carcinogenesis and progression of NSCLC. The up-regulation of Notch1, Jagged1 and VEGF protein expression probably predict NSCLC carrying relatively strong permeation and metastasis.
Adult ; Aged ; Calcium-Binding Proteins ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Jagged-1 Protein ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; Neoplasm Staging ; Receptor, Notch1 ; metabolism ; Serrate-Jagged Proteins ; Vascular Endothelial Growth Factor A ; metabolism

Objective To optimize ambi-extracting and inclusion process of volatile oil from Chuanxiong Rhizoma and Angelicae Sinensis Radix. Methods With yield ratio of volatile oil and ferulic acid content in water extract as evaluation indexes, single factor experiments were used to study the extraction process. With the inclusion rate of volatile oil and yield of inclusion as evaluation indexes, saturated aqueous solution was used to L9(34) orthogonal experiments to reach optimum inclusion process. Results The optimum extraction process of Chuanxiong Rhizoma and Angelicae Sinensis Radix was extracted for 8 hours with 8 folds the amount of water, and without soaking. The validation experiments of extraction of volatile oil and ferulic acid content in water extract were 1.23 mL and 0.387 9 mg/g. The optimum conditions of inclusion process were as follows: volatile oil (mL): β-CD (g) was 1:8;inclusion temperature was 40 ℃; inclusion time was 3 hours. The validation experiments of inclusion rate of volatile oil and yield of inclusion were 74.89% and 72.81%. Conclusion Optimum ambi-extracting and inclusion process of volatile oil from Chuanxiong Rhizoma and Angelicae Sinensis Radix are feasible and stable, witch can provide certain supporting data for preparation production.

BACKGROUNDTo investigate the dynamics of amyloid fiber formation of yeast (Saccharomyces cerevisiae) prion protein Sup35NM under the native condition to provide materials and clues for the elucidation of amyloid fiber formation.

METHODSThe Sup35NM gene was cloned and expressed in E. coli. The recombinant Sup35NM protein was purified under denaturing conditions through Nickel-Sepharose chromatography. Aliquots were removed at designated time points for transmission electron microscopy (TEM), circular dichroism (CD) spectra, protease K resistance assay, as well as thioflavin T (ThT) binding assay.

RESULTSThe Sup35NM expressed and purified under denaturing conditions. The morphological alteration of the Sup35NM in PBS (pH7.4) during the protein aggregation and amyloid fiber formation was visualized by TEM. The CD assay showed that the course of amyloid fiber formation underwent a conformational shift from alpha-helix to beta-sheet. The fibers had higher capacity of resistance to protease K digestion compared to the monomers. ThT fluorescence assay displayed a rapid growth phase before reaching a final equilibrium phase during the fiber formation, and the higher concentration of Sup35NM could greatly accelerate the fiber formation in vitro.

CONCLUSIONYeast prion protein Sup35NM forms amyloid readily under native conditions in vitro. The dynamics of Sup35NM amyloid formation may provide supporting evidences for the nucleating polymerization models of amyloid fiber formation.

Amyloid beta-Peptides ; genetics ; metabolism ; ultrastructure ; Electrophoresis, Polyacrylamide Gel ; Endopeptidase K ; metabolism ; Kinetics ; Microscopy, Electron ; Peptide Termination Factors ; Prions ; genetics ; metabolism ; ultrastructure ; Protein Binding ; Recombinant Proteins ; metabolism ; ultrastructure ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; metabolism ; ultrastructure ; Thiazoles ; metabolism

Microgravity is known to produce a number of neurological disturbances during space flight; however, the underlying mechanism of these disturbances is yet to be elucidated. There have been some reports about the increased oxidative stress under microgravity or simulated microgravity. In the present study, we investigated the process of oxidative stress induced by simulated microgravity in different areas of rat brain, which may shed light on the mechanism of neurological disturbances and further neuroprotective research in spaceflight. After adaption for 7 d, 40 healthy male Sprague-Dawley rats were matched for body weight and randomly assigned to control groups (7, 14, 21 and 28 d) and tail-suspended simulated microgravity groups (7, 14, 21 and 28 d). The tail-suspended groups were treated with 30 angels of tail suspension and the control groups were treated similarly to the tail-suspended groups but without tail suspension. After the required times, different structures of rat brain, including cerebellum, cerebral cortex and hippocampus, were harvested and frozen for the further determination. Griess assay, thiobarbituric acid reactive substance (TBARS) assay, competitive ELISA and ferric reducing ability of plasma (FRAP) assay were used for the observation of the changes of reactive nitrogen species (RNS), malondialdehyde (MDA), nitrotyrosine (NT) and total antioxidant capacity (TAC), respectively. As shown in the results, there were different changes in various brain regions after tail suspension compared with control groups. (1) In cerebellum, NT increased after 7 d tail suspension, decreased after 14 d and increased again after 28 d; MDA increased after 14 d; RNS increased and TAC decreased after tail suspension for 21 d; (2) Increase of NT after14 d tail suspension, increase of MDA and decrease of TAC after 21 d were found in cerebral cortex; (3) In hippocampus, RNS increased after tail suspension for 7 d, decreased after 14 d and increased again after 28 d; MDA increased after 21 d; NT increased after 28 d; TAC increased after 7 d and recovered after 21 d. These results suggest that simulated microgravity induced by tail suspension increases the level of oxidative stress in rat brain; however, there are different features in different areas of rat brain. During the response to simulated microgravity, rat brain tissues present a similar process from adaptive response to irreversible oxidative damage.
Animals ; Antioxidants ; metabolism ; Brain ; physiopathology ; Hindlimb Suspension ; Male ; Malondialdehyde ; metabolism ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Weightlessness Simulation

OBJECTIVETo elvaulate the antioxidant activity of the pigment of Lycium ruthenicum.

METHODThe antioxidant activities were measured by the effects of the reducing ability, scavenging DPPH. H2O2-induced hemolysis of mice erythrocyte, serum resistance of reactive oxygen species, content of MDA in liver tissue, and swelling effect of mitochondria in liver tissue.

RESULTThe pigment of L. ruthenicum could scaveng DPPH* remarkably with IC50 0.164 mg x mL(-1), inhibitte hemolysis of mice erythrocyte evidently with IC50 0.112 mg x mL(-1). The resistant of reactive oxygen species was enhanced by the tested substances, simultanously. The concentration of MDA of peroxidation of lipid in mice liver could be reduced, and the swelling of mice liver mitochondria alse be restrained.

CONCLUSIONThe pigment of L. ruthenicum has antioxidant activity in tested concentration.

Animals ; Antioxidants ; pharmacology ; Biphenyl Compounds ; metabolism ; Erythrocytes ; drug effects ; Free Radical Scavengers ; pharmacology ; Hemolysis ; drug effects ; Hydrazines ; metabolism ; Lipid Peroxidation ; drug effects ; Liver ; metabolism ; Lycium ; chemistry ; Malondialdehyde ; metabolism ; Mice ; Mitochondria, Liver ; pathology ; Mitochondrial Swelling ; drug effects ; Phenols ; isolation & purification ; pharmacology ; Picrates ; Pigments, Biological ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Reactive Oxygen Species

OBJECTIVETo establish an in vitro model of mouse cochlear basilar membrane impairment using cisplatin, and observe the protective effect of methionine on the hair cells.

METHODSThe cochlear basilar membrane samples of thirty two Kunming mice were harvested on the 2nd day after birth and randomly divided into four groups. Each group had 16 samples. Overnight preincubation the cochlear organ followed by appropriate treatment respectively as follows: the serum-free culture medium, the serum-free culture medium with methionine and cisplatin, the cisplatinum-containing serum-free culture medium, and the methionine-containing serum-free culture medium. The protective effect of methionine for injury of cochlea hair cells induced by cisplatin was observed by myosin-VI immunofluorescence, light microscopy, laser confocal scanning microscope and hair cells counting.

RESULTSThe outer hair cells (OHC) and inner hair cells (IHC) of control group and methionine group were not damaged. The outer and inner hair cells of cisplatin group were damaged in various degree, and had remarkable difference compared with control group and methionine group (P < 0.05). The outer hair cells and inner hair cells of cisplatin + methionine group were damaged less than the cisplatin group with remarkable difference (t(IHC) = 3.929, t(OHC) = 8.582, P < 0.05).

CONCLUSIONSCisplatinum could damage the cochlear hair cells of the basal membrane in Kunming mice. Methionine might protect against cisplatin's damage on the cochlear hair cells.

Animals ; Cisplatin ; adverse effects ; pharmacology ; Hair Cells, Auditory ; drug effects ; In Vitro Techniques ; Methionine ; pharmacology ; Mice ; Mice, Inbred Strains