Objective: To design a new type of maintenance learning machine of ventilator for teaching and experiment combined with electronic technology course of higher vocational education. Methods: ACM804 ventilator was chosen as prototype machine and the electronic technology course of higher vocational education was chosen as theory basis. The designs of internal circuit and gas circuit of prototype machine were improved, besides, some test points and failure points were designed for teaching in classroom. Results: This machine not only could realize all functions of prototype machine, but also could carry forward test teaching and contribute students to maintain common failure in accordance with course content. Conclusion: Through the design of teaching experiment and using of learning machine, the new machine has achieved better teaching effect for students to grasp machine theory and operate actual circuit.

0.05).Both HMGB1 mRNA expression and HMGB1 protein level were remarkably higher in LPS treatment group than that in control group(P0.05).CONCLUSION:RPM inhibits HMGB1 expression not only by directly suppressing STAT3 activation,but also by indirectly reducing TNF-? level.

BACKGROUND:Vaginal smooth muscle cells are mainly cultured by enzyme digestion method and explant method.The former method requires a short culture time with a high output,but this method demands a large number of tissues with a high opportunity of pollution,and the optimal digestion time is difficult to control.The latter method is simple and effective,but the culture needs a long time.OBJECTIVE:To observe the culture time of rabbit vaginal smooth muscle cells using explant + enzyme digestion methods,and to compare with the explant method.DESIGN,TIME AND SETTING:The in vitro observation experiment was performed at the Burn Institute and Animal Laboratory of First Affiliated Hospital,Nanchang University from February 2005 to February 2006.MATERIALS:Female New Zealand rabbits aged 4-5 months,with the body mass of 2.0-2.5 kg were selected for this study.METHODS:Vaginal smooth muscle cells using explant method:1 mm?1 mm?1 mm explants were uniformly incubated in a culture medium,and then placed in a incubator at 37 ℃ for 3.0-4.0 hours.After tissue confluence,tissues were immersed in DMEM containing 0.1 volume fraction,stored at 37 ℃ for 3.0-4.0 days.Following cells grew from the tissue islets,the medium was changed about once every three days.Vaginal smooth muscle cells using explant + enzyme digestion methods:The tissue pieces were digested by 0.1% collagenase type Ⅳ at 37 ℃ for 0.5-1 hour,and then enzymatic digestion was interrupted when the edge of tissue pieces became coarse.These pieces were cultivated in culture dishes.The remaining steps were the same as the explant method.Serial subculture:The 1st subculture was made when 50%-60% cells were confluent.After the 2nd passage of cells was obtained,subculture was made when 80% cells were confluent.MAIN OUTCOME MEASURES:Time of primary culture;Surface structure and ultrastructure of the 3rd passage of vaginal smooth muscle cells.RESULTS:The eruption time and the confluent time of vaginal smooth muscle cells could be shortened by the explant + collagen digestion methods about 1-2 days and 3-4 days respectively compared with the explant method only.Vaginal smooth muscle cells,which were obtained by the two methods mentioned above,could propagate for 5-6 passages.The attached cells were polygonal or spindle and after confluence could be seen,the marked smooth muscle cells "Peak-Valley" structure in some domains under the inverted microscope.The third passage of smooth muscle cell nuclei were serration;the cytoplasm contained the marked smooth muscle cell structural myofilaments and dense bodies and plenty of golgi bodies;the rough endoplasmic reticulum broadened;the free ribosome were abundant under the transmission electron microscope.This indicated that vaginal smooth muscle cells with synthesis phenotype could be greatly collected.CONCLUSION:Explant + enzyme digestion methods need a short cycle of primary culture.Vaginal explants should be kept moisture during drawing materials,isolation and the whole.The time of enzymatic digestion should not be too long,to the point that palpi pilosi is seen surrounding the explants.Primary culture should remain static state and use of plastic culture dishes.

AIM: To investigate the correlation of intestinal endotoxemia (IETM), histaminemia and cellular immune function in the patients with hepatitis B. METHODS: Peripheral blood was collected from patients with chronic hepatitis B (n=80) and healthy individuals (n=18). According to plasma endotoxin concentration, total patients were divided into two groups: ET positive and ET negative. Serum IL-10, IL-12, IFN-?, IL-2, IL-4 concentrations were detected. In addition, the serum histamine (HA), tryptase (TS) and AP50 levels were studied. RESULTS: Compared to control group, the concentrations of IL-4 and IL-10 were increased, but IL-12 and IFN-? were decreased obviously in total patients (P

AIM: To investigate that nicotine inhibits HMGB1 expression and release in RAW264.7 cells.METHODS: (1) RAW264.7 cells were cultured in 6 wells plate, treated with 250 μg/L LPS and 1 μmol/L or 10 μmol/L nicotine, in which the cells treated with or without 250 μg/L LPS were regarded as nicotine 1 group (N1), nicotine 2 group (N2), LPS group (LPS) and control group (C), respectively. HMGB1 protein in the cell culture media and in cell nuclear was examined by Western blotting and the cellular HMGB1 mRNA level was detected by RT-PCR. (2) Transfected with antisense RNA or sense RNA of α~7 subunit-containing nicotinic receptor (α~7nAChR), RAW264.7 cells were treated with 250 μg/L LPS and 10 μmol/L nicotine, HMGB1 protein in the culture media was also tested by Western blotting.RESULTS: (1) HMGB1 mRNA level in C group was low (1 659.20±121.05) and no significant statistical difference among groups of N1, N2 and LPS was observed (P>0.05). (2) Higher HMGB1 accumulation in the cell culture media was detected in LPS group (445.34±28.52) than that in C group. Compared to LPS group, both N1 and N2 groups distinctly attenuated HMGB1 accumulation in culture media (P<0.05). (3) Nuclear HMGB1 accumulation was lower in LPS group than that in C group, and two different nicotine concentrations markedly increased the nuclear HMGB1 accumulation compared to LPS group (P<0.05). (4) No significant difference of HMGB1 levels in culture media between antisense RNA group and LPS group was observed (P>0.05). In sense RNA group, however, HMGB1 level was observably reduced compared to antisense group (P<0.05).CONCLUSION: The present results suggest that nicotine dramatically inhibits RAW264.7 cell nuclear HMGB1 translocation and extracellular release, and this effect relies on α~7nAch receptor expression.

Aim To investigate the anti-neuroinflam-matory activities of dehydromiltirone and the underlying mechanisms in LPS-stimulated microglial cell line BV2 cells. Methods BV2 cells were pre-treated with de-hydromiltirone, then stimulated by LPS. The levels of nitric oxide( NO) were measured by Griess assay, and the concentrations of pro-inflammatory cytokines were measured by ELISA assay. Confocal fluorescence mi-croscopy was used to measure the expression of MAC-1, the biomarker of activated BV2 cells. The levels of-inducible nitric oxide synthase ( iNOS ) , cyclooxygen-ase-2 ( COX-2 ) , NF-κB and PI3 K/Akt were deter-mined by Western blot analysis. Results The treat-ment of dehydromiltirone significantly inhibited the pro-duction of NO, TNF-α and IL-6, attenuated the ex-pression of iNOS and COX-2 protein, and dampened the microglial activation in LPS-stimulated BV2 cells. The mechanistic study revealed that dehydromiltirone inhibited the phosphorylation of PI3 K and Akt in LPS-stimulated BV2 cells, and decreased NF-κB activation by suppressing the degradation of IκB. Conclusion dehydromiltirone shows significant anti-neuroinflamma-tory effects through inhibiting PI3 K/Akt phosphoryla-tion and then inhibiting NF-κB signaling pathway.

OBJECTIVETo observe the changes in expression of microRNA-203 and P63 in human epidermal stem cells and KCs, and to investigate their effects and significance in the epidermal proliferation and differentiation.

METHODS(1) Five normal foreskin tissue specimens were collected from 5 patients by circumcision in Department of Urinary Surgery of the First Affiliated Hospital of Nanchang University from March to June in 2013. Then single cell suspension was obtained by separating epidermis with trypsin digestion method. The cells were divided into quick adherent cells and non-quick adherent cells by type IV collagen differential adherent method. The biological characteristics of cells were observed by inverted phase contrast microscope immediately after isolation and on post culture day (PCD) 3. The expression of CD29, keratin 19, keratin 1, and keratin 10 was identified by immunocytochemical staining. The expression of microRNA-203 and mRNA of P63 was determined by real-time fluorescent quantitative RT-PCR. The protein expression of P63 was determined by Western blotting. Data were processed with t test and Pearson correlation analysis.

RESULTS(1) Immediately after isolation, quick adherent cells were small, round, and dispersed uniformly. On PCD 3, the cells adhered firmly, and they grew in clones. Immediately after isolation, non-quick adherent cells appeared in different shapes and sizes, and dispersed unevenly. On PCD 3, the cells adhered precariously and did not show clonal growth. Quick adherent cells showed positive expression of CD29 and keratin 19, while non-quick adherent cells showed positive expression of keratin 1 and keratin 10. Quick adherent cells were identified as epidermal stem cells, and non-quick adherent cells were identified as KCs. (2)The expression level of microRNA-203 in epidermal stem cells (0.74 ± 0.20) was lower than that in KCs (3.66 ± 0.34, t =16.582, P <0.001). The mRNA expression level of P63 in epidermal stem cells (4. 16 ± 0.28) was higher than that in KCs (2.90 ± 0.39, t =5. 850, P =0.001). The protein expression level of P63 in epidermal stem cells (1.42 ± 0.05) was higher than that in KCs (0.73 ± 0.03, t =26.460, P <0. 001). (3) The expression level of microRNA-203 was in significantly negative correlation with the expression levels of mRNA and protein of P63 (with r values respectively - 0. 94 and -0.98 , P values below 0.05).

CONCLUSIONSThe expression levels of microRNA-203 and P63 in human epidermal stem cells and KCs were significantly different, which might be related to the different characteristics of proliferation and differentiation of the cells.

Cell Differentiation ; Cells, Cultured ; Epidermis ; cytology ; growth & development ; Epithelial Cells ; cytology ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Humans ; Integrin beta1 ; Keratin-10 ; genetics ; metabolism ; Keratin-19 ; genetics ; metabolism ; Keratinocytes ; Male ; Membrane Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; metabolism ; Stem Cells ; cytology ; metabolism

AIM: The goal of the present study was to evaluate the possibility about enterogenous endotoxemia in pathogenesis of hepatopulmonary syndrome. METHODS: The rat model of cirrhosis was prepared with compound factors. A small dose of lipopolysaccharide (LPS) was administered intraperitoneally once to aggravate endotoxemia of animal with cirrhosis. The normal rats injected with LPS or injected with LPS combined with glycine (LPS antagonist) were designed as controls. RESULTS: Hepatopulmonary syndrome of rats with cirrhosis had occurred in the end of eighth weeks. Pulmonary pathological changes of cirrhosis rats were exacerbated after administration of a given dose of LPS. Glycine sharply antagonized the biological effect of LPS in vivo and in vitro, inhibited the production of TNF-? by LPS and alleviated various pathological changes of hepatopulmonary syndrome. CONCLUSIONS: Enterogenous endotoxemia in cirrhosis rats might be an important mechanism in the development of hepatopulmonary syndrome. Endotoxin and its mediating effect by way of cytokines (TNF-?) may play a role in the pathogenesis of hepatopulmonary syndrome.

OBJECTIVETo investigate the isoflavones from the vines of Pueraria lobata.

METHODThe compounds were isolated by column chromatography over silica gel and RP-C18, and purified by Sephadex LH-20 column chromatography and preparative TLC. The structures were elucidated on the basis of physico-chemical properties and spectral data.

RESULTTwelve compounds were isolated and identified as: 3'-methoxydaidzein (1), formononetin (2), genistein (3), daidzein (4), daidzin (5), genistin (6), ononin (7), 5-hydroxyl ononin (8), calycosin (9), 6"-O-acetyl genistein (10), 6"-O-acetyl daidzin (11), puerarin (12).

CONCLUSIONFor the first time, compounds 9-11 were isolated from the genus Pueraria plant, and compounds 1, 3, 6-8 were obtained from the vines of this plant.

Genistein ; chemistry ; Glucosides ; chemistry ; Isoflavones ; chemistry ; Magnetic Resonance Spectroscopy ; Plant Stems ; chemistry ; Pueraria ; chemistry

ABSTRACT:Objective To explore the role of HIF‐1αin the pathogenesis of hepatopulmonary syndrome (HPS) and its relationship with GRP78 .Methods The HPS model in rats was induced by multiple pathogenic factor .The samples were assessed by using Western blotting analysis for HIF‐lα, GRP78 and VEGF164 . The expressions of VEGFR‐2 and CD105 were observed by using immunohistochemical staining .Results The protein level of HIF‐1αwas significantly increased in HPS group at week 8 compared with that at week 4 and 6 groups and corresponding normal control groups .With the development of HPS ,protein level of GRP78 was gradually increased at each time point significantly and reached the highest level at week 8 ;protein level of VEGF164 showed a similar change with GRP78 ,but the peak was at week 6 .Immunohistochemical results showed that the protein expressions of VEGFR‐2 and CD105 were gradually increased in lung tissue as HPS progressed .The protein level of GRP78 was positively correlated with HIF‐1α,VEGF164 ,VEGFR‐2 and CD105 ,respectively (P<0 .05) .Conclusion HIF‐1αis most likely together with GRP78 to play a critical role in promoting pulmonary microvascular remodeling in the pathogenesis of HPS in rats .