BACKGROUND: Intestinal mucosa was damaged by ischemia and hypoxia during severe burn, and injury and infection of oxygen-derived free radicals during reperfusion.OBJECTIVE: To study on the basis of sub-cell level whether danshen can improve respiratory function of mitochondrion of epithelial cell in intestinal mucosa and decrease the production of oxygen-derived free radicals so as to protect intestinal mucosa of burnt rats.DESIGN: Randomized controlled animal study with single sample.SETTING: Department of Burn, the Third Hospital of Chinese PLA.MATERIALS: The experiment was conducted at the Surgery Institute of the Third Hospital of Chinese PLA from December 2001 to February 2002.Totally 96 SD rats of clean grade were randomly divided into normal control group (n=24), burning model group (n=36) and danshen treatment group (n=36). 1 mL danshen parenteral solution had 1.5 g raw materials.METHODS: ① Rats in the burning model group and danshen treatment group were used to establish grade Ⅲ burning models with 20% scald of total body surface. ② Rats in the normal control group were not injured.③ After modeling, 1 mL/kg danshen parenteral solution was slowing injected into rats in the danshen treatment group through femoral vein, but saline was injected slowly into rats in the normal control group and burning model group. ④ Twelve rats from each of the burning mgdel group and the danshen treatment group were sacrificed at 1, 2 and 6 hours after modeling, and 8 rats in the normal control group were sacrificed at relevant time points respectively. ⑤ Samples of small intestine were collected to measure cytochrome aa3, cytochrome C, level of energy charge and activity of superoxide dismutase of mitochondrion of epithelial cells in intestinal mucosa.MAIN OUTCOME MEASURES: Cytochrome aa3, cytochrome C, level of energy charge and activity of superoxide dismutase of mitochondrion of epithelial cells in intestinal mucosa 1, 2 and 6 hours after modeling.RESULTS: Totally 96 SD rats entered the final analysis. ① Level of cytochrome aa3 of mitochondrion of epithelial cells in intestinal mucosa at various time points after modeling: Levels of cytochrome aa3 did not changed obviously in the burning model group and danshen treatment group 2 hours after modeling as compared with that in the normal control group (P＞ 0.05). Six hours after burning, levels in the burning model group were obviously lower than those in the normal control group (P ＜ 0.05), but those in the danshen treatment group were obviously higher than those in the burning model group [(3.16±0.13), (2.5640.15) μkat/g, P ＜ 0.05]. ②Measurements of cytochrome C, level of energy charge and activity of superoxide dismutase of mitochondrion of epithelial cells in intestinal mucosa at various time points after modeling: As compared with those in the normal control group, measurements were decreased obviously in the burning model group 1, 2 and 6 hours after modeling (P ＜ 0.05 or 0.01), but those in the danshen treatment group were obviously higher than those in the burning model group (P ＜ 0.05 or 0.01).CONCLUSION: Danshen can raise the levels of cytochrome aa3, cytochrome C, energy production and SOD, and also reduce the production of oxygen-derived free radicals so as to improve respiratory function of mitochondria of epithelial cells in intestinal mucosa of rats.

BACKGROUND:Previous studies have mainly focused on costal cartilage, articular cartilage, nasal septal cartilage, and auricular cartilage, but in vitro culture of human vertebral endplate cartilage is stil rarely reported. OBJECTIVE: To discuss the feasibility of culture of vertebral endplate chondrocytes from type I neurofibromatosis associated with scoliosis patientsin vitro and to study the biological characters of the chondrocytes. METHODS: Through two-step enzymatic digestion and tissue culture, the chondrocytes from the vertebral endplate of seven type I neurofibromatosis patients isolated and cultured in monolayer and passaged to observe the changes of cel morphology under inverted phase contrast microscope. Colagen type II expression was detected by immunocytochemistry to identify whether the cels had chondrocyte characters. The growth kinetics was detected by using MTT colorimetric assay to draw the growth curve of passage 2 chondrocytes. RESULTS AND CONCLUSION:A few chondrocytes crawled from the cartilage after 2 weeks culture and cels were passaged at 3 weeks. Along with passage going on, the phenotype of chondrocytes was changed from polygonal, round, triangle, and irregular shapes to fusiform. The colagen type II expression in passage 2 cels was positive by immunohistochemical staining. MTT test showed the growth curve of the passage 2 chondrocytes presented a transverse “S”. Cels were found logarithmic growth at days 4-7, reached platform stage at days 8-13, and decreased at day 14. It is an effective and simple procedure by two-step enzymatic digestion and tissue explant method to culture vertebral endplate chondrocytes with high purity and good viability from type I neurofibromatosis patients associated with scoliosisin vitro. Passage 2 chondrocytes from the vertebral endplate exhibit the best viability at days 4-7, which can be used as targets for research of pathogenesis of type I neurofibromatosis with atrophic scoliosis.

Objective: To study the effect of salvia miltiorrhiza on mitochondria respiratory function of hepatic cells and oxygen radicals in rats with burn.Methods: Ninety-six Sprague-Dawley rats were randomly divided into normal control group,model group and salvia miltiorrhiza group.Rats in model group and salvia miltiorrhiza group were subjected to a 20% total body surface area(TBSA) full-thickness thermal injury.The levels of cytochrome aa3,cytochrome C,energy production and superoxide dismutase(SOD) in mitochondria of hepatic cells were dynamically determined at 1,2 and 6 hours after injury.Results: ①The levels of cytochrome aa3 in model group and salvia miltiorrhiza group did not change obviously within 2 hours after burn,while they were much lower than that in normal control group at 6 hours after burn,and the level of cytochrome aa3 in salvia miltiorrhiza group was obviously higher than that in model group(all P

Aim To investigate the protective effects of Diterpene Ginkgolides Meglumine Injection(DGMI)on SY5 Y cells damaged by oxygen-glucose deprivation and its functional mechanisms.Methods After 4 h of OGD,the cells were treated with 25 mg·L-1 drugs for 1 h.Subsequently,cell viabilities were measured by cell counting kit-8(CCK-8 kit)and cell apoptosis was measured by flow cytometric analysis.Furthermore, the mitochondrial membrane potential was detected by rhodamine123 staining.The levels of phospho-p38, phospho-p53,Bcl-2,Bax and cleaved caspase-9/3 were evaluated by western blot.Results DGMI signif-icantly increased the cell viabilities of SY5 Y cells dam-aged by OGD,and reduced OGD-elicited dissipation of mitochondrial membrane potential and cell apoptosis. Furthermore,DGMI also reduced p-p38,p-p53,Bax/Bcl-2 ratio,cleaved caspase-9 and cleaved caspase-3. Conclusion DGMI shows good neuroprotective effects on SY5 Y cells after oxygen-glucose deprivation.The underlying mechanisms may be associated with the sup-pression of p38/p53/Bcl-2 /caspase-9/caspase-3 sig-naling pathway.

OBJECTIVE To investigate the protective effects and mechanism of diterpene ginkgolides meglumine injection (DGMI) against oxidative stress induced by oxygen-glucose deprivation (OGD) in SH-SY5Y cells. METHODS SH-SY5Y cells were divided into five groups: normal control, model control (OGD group) and drug(25 mg · L- 1) administration groups including DGMI group, extract of ginkgo biloba leaves injection group (EGBLI) and lactones ginkgo biloba injection group (LGBI). The cells suffered from oxygen-glucose deprivation (OGD) for 4 h, followed by reoxygenation with drugs for 6 h. Then, cell viabilities were detect using CCK-8 assays, reactive oxygen species (ROS) levels using fluorescence probe DCFH-DA and superoxide dismutase (SOD) activities using WST-1 test. Western blotting was used to detected protein levels of hemeoxygenase-1(HO-1), NAD(P)H, quinone oxidore?ductase l (Nqo1), protein kinase B (Akt), phosphorylated Akt (p-Akt), nuclear factor-E2-related factor2 (Nrf2) and phosphorylated Nrf2 (p-Nrf2). The cells were induced by OGD for 4 h, followed by reoxygen?ation and DGMI for 1 h, combined with different concentrations of PI3K inhibitor (LY294002) (at the final concentration of 12.5, 25 and 50 μmol · L-1) before the protein levels of AKT, p-AKT, Nrf2 and p-Nrf2 were detected by Western blotting. RESULTS SH-SY5Y cells induced by OGD for 4 h resulted in an increase in ROS(P<0.01), but a decrease in cell viabilities(P<0.01), SOD activities(P<0.01), and antioxidant protein levels ( Akt, p-Akt, Nrf2, p-Nrf2, HO-1 and Nqo1) (P<0.01). Compared with OGD group, treatment with reoxygenation and drugs (DGMI,EGBLI and LGBI respectively) for 6 h resulted in a decrease in ROS (P<0.01), but an increase in cell viabilities, SOD activities and antioxidant protein levels of p-Nrf2, HO-1, Nqo1 and p-Akt(P<0.05,P<0.01). DGMI group showed the best efficiently. Moreover, after OGD for 4 h, compared with DGMI group, combining reoxygenation and DGMI with LY294002 for 1 h resulted in a concentration-dependent inhibition of the protein levels of p-AKT and p-Nrf2(P<0.01). CONCLUSION DGMI 25 mg · L-1 can inhibit oxidative stress in SH-SY5Y cells induced by OGD by increasing the activity and expression of Nrf2 through PI3K/Akt pathway, which may be one of the mechanisms by which DGMI protects neurons from stroke.

Aim To investigate the protective effects of YXETNZ injection on SH-SY5 Y cells damaged by oxygen-glucose deprivation ( OGD ) , and explore its functional mechanisms. Methods After 4 h of OGD, the cells were treated with 25 mg·L-1 drugs for 1 h. Subsequently, cell viabilities were measured by cell counting kit-8 ( CCK-8 kit ) and cell apoptosis was measured by caspase-3/7 assay kit according to manu-facturer’ s instructions. Furthermore, cell death was also detected by ELISA. The levels of phospho-Akt, phospho-PKA,phospho-Bad were evaluated by western blot. Results Oxygen-glucose deprivation significant-ly decreased the cell viabilities of SH-SY5Y cells, while YXETNZ injection significantly increased cell vi-abilities, phospho-Akt, phospho-PKA and phospho-Bad. Furthermore, YXETNZ injection also reduced the activities of caspase-3/7 and cytoplasmic histone-asso-ciated-DNA-fragments contents. Conclusion Our re-searches demonstrat that YXETNZ injection shows good neuroprotective effects on SH-SY5 Y cells after oxygen-glucose deprivation. The underlying mechanisms may be associated with activation of PI3 K/Akt/Bad/caspase-3/7 , cAMP/PKA/Bad/caspase-3/7 signaling pathway.

Objective To analyze functioning and disability, unmet needs and the service for people with disabilities in Shenzhen, Guangdong, China. Methods Based on International Classification of Functioning, Disability and Health (ICF) and World Report on Disabil-ity, the theoretical framework had been constructed, and the unmet needs and service status for people with disabilities in Shenzhen in 2015 had been statistically analyzed. Results For the statistics of disability by categories, physical disability composed of 42.5%, speech disability composed of 1.5%;people with severe and extremely severe disabilities composed of 52.8%;Futian District composed of 19.7%, Yantian District composed of 2.2%;people aged 0 to 18 years composed of 17.4%, and people aged over 60 years (27.0%) were the larger group. For the unmet needs of people with disabilities, 25%needed rehabilitation therapy, 18.8%needed functional training, 23.2%needed assis-tive devices, and 32.9%had no need. For rehabilitation sevice in Shenzhen, 24.4%received rehabilitation therapy, 17.4%received function-al training, 20.4%received assistive devices, and 37.6%did not receive any service. For the barrier-free reconstruction, 4.6%needed bath-room reconstruction, and 0.7%needed internet access screen software. Conclusion The status of functioning and disability, unmet needs and service development of rehabilitation in Shenzhen had been analyzed. There was still a gap between unmet needs and services of rehabilita-tion. It recommended to construct precise services delivery based on unmet needs, improve the full coverage and quality of service of reha-bilitation.

To investigate the anti-apoptotic effect of diterpene ginkgolides meglumine injection(DGMI)on SH-SY5Y cells induced by oxygen-glucose deprivation/reoxygenation(OGD/R), and to explore its mechanisms. After 4 h of OGD, the SH-SY5Y cells were treated with 25 mg/L DGMI for 1 h. The release of lactic dehydrogenase(LDH)was measured by cytotoxicity detection kitplus. Cell apoptosis was detected by caspase-3/7 assays. Cell death was detected by ELISA. The concentration of ［Ca2+］i in cytoplasm was measured by Fluo-3 AM and the levels of calpain and cleaved capaease-12 were evaluated by western blot. As we expected, DGMI significantly decreased the release of LDH, the concentration of ［Ca2+］i, the protein levels of calpain and cleaved caspase-12. Furthermore, DGMI injection also attenuated the activities of caspase-3/7 and the contents of cytoplasmic histone-associated- DNA-fragments. These data demonstrated that the DGMI injection showed good anti-apoptotic effect in SH-SY5Y cells induced by OGD/R. The mechanisms may be associated with the inhibition of Ca2+/calpain/caspase-12/caspase-3 signaling pathway.

Objective To develop a novel transcatheter tricuspid valve replacement device and test its performance. Methods The transcatheter tricuspid valve stent consisted of double-layer self-expanding nitinol stent, biotissue-derived bovine pericardial leaflets, and PTFE woven. The delivery system, mainly consisting of a handle control unit and a delivery sheath, was sent to the correct position via right atrium or jugular vein. The sheath had a visualization feature, and the handle control unit could realize the functions of stable release and partial recovery of the interventional valve. In addition, this study performed animal survival experiments on the basis of in vitro experiments. A large-white pig was used as the experimental animal. Cardiopulmonary bypass was established through median thoracotomy, then the right atrium was opened, and the interventional valve was released under direct vision without cardiac arrest. Approximately 1 month after interventional valve implantation, the maneuverability and stability of the interventional tricuspid device were evaluated by autopsy. Results Through the animal experiment, the interventional valve was successfully released, and the anchoring was satisfactory. Postoperative transthoracic echocardiography showed that the interventional valve opened and closed well, the flow rate of tricuspid valve was 0.6 m/s, and there was no obvious tricuspid regurgitation. One month after the operation, we dissected the large-white pig and found the interventional valve was not deformed or displaced, the leaflets were well aligned, and there was thrombus attachment in the groove between the inner and outer layers of the interventional valve. Conclusion Animal experiment shows that the novel device can stably and firmly attach to the tricuspid annulus, with good anchoring effect, and effectively reduce paravalvular leakage.