Objective To investigate the effect of ?,?-carotene in enhancing the function of anti-oxidative damage in osteoblasts.Methods M3T3-E1 osteoblasts were randomly divided in five groups: control group,model group,and ?,?-carotene group [low-dose(1?10-8mol/L),medium-dose(1?10-7mol/L),high-dose(1?10-6mol/L)].The activity of cells,superoxide dismutase(SOD),the content of reactive oxygen species(ROS),lipid oxygen(LPO),and membrane fluidity were tested and compared. Results Compared with those in ?,?-carotene groups,the activity of cells,SOD activity and membrane fluidity in the model group were significantly decreased(P

This article was aimed to explore theeffect of Pi-Zhen(PZ) on 5-hydroxytryptamine(5-HT) in nerve end-ings tension painrabbitmodel. Forty-two healthy male big ear whiterabbits of three months old were randomly divided into 5 groups, which were the normal group,model group, drug group, Hao-Zhen (HZ) group, and PZ group. Nerve endings tension pain modelswere established. No operation was made in the normal group. The amount of 1 ml saline was injected to the operative sitebetween the shallow and deep fascia in each rabbit and then draw out at 5, 10, 20, 30, 40, 50, 60 min after operation. The 5-HT level in extracting solution of different groups was determined with ELISA method. The results showed that PZ interventioncan reducethe 5-HT level ofextracting solution in operative site; and with the increasing of time, the level of 5-HT was gradually reduced. Compared with the model group, the contents of 5-HT in the treatment group were significantly decreased at different time points(P<0.01). The content of 5-HT in the PZ group was significantly higher than that of the normal group at 5 min (P<0.01). Compared with the drug group, the contents of 5-HT in the HZ group and PZ group were significantly increased at 5, 10, 20, 30, 40 min (P<0.01). Compared with the normal group, the contents of 5-HT in the PZ group was significantly decreased at 40, 50, 60 min (P<0.05). Compared with the HZ group, the contents of 5-HT were significantly decreased at 20, 30, 40, 50, 60 min (P<0.01). There was no difference at other time points. It was concluded that PZ can ease pain through reducing the tension of local soft tissues, decreasing 5-HT releasing, and promoting its degradation.

Objective To investigate the stimulation of exosome derived from osteosarcoma cells suppressing cytotoxic T cells and the inhibitory effect of active T cells for surpressing osteosarcoma cells. Methods Exosome derived from tumor cells was isolated and purified by ultracentrifugation. Its morphology was observed with transmission electron microscope, and the major histocompatibility complex-I ( MHC-I) molecules were analyzed by western blot. Mice bone marrow-derived dendritic cells were pulsed with exosome. Surface membrane MHC-I molecules were analyzed with flow cytometry. The effect of active T cells on the growth of osteosarcoma cells were detected by MTT assay after the T cells being stimulated by exosome-pulsed dendritic cells. Results The exosome was round or near round corpuscle, and the diameter was about 50-100 nm by transmission electron microscope. The size was relatively homogeneous. Western blot showed that the exosome expressed MHC-Ⅰmolecules. Surface membrane CD80,MHC-I and II molecules were expressed on 77. 16%, 83. 21%, and 91. 26% of LPS-treated dendritic cells, respectively, which were up-regulated compared to untreated cells. Dendritic cells pulsed with exosome derived from osteosarcoma cells caused significantly higher T cells stimulation and osteosarcoma cells inhibition as compared to un-pulsed dendritic cells. (P<0.05). Conclusion T cells can inhibit the growth of osteosarcoma cells after being stimulated by exosome-pulsed dendritic cells in vitro.

AIM:To investigate the ability of a metal complex ammonium tetrathiomolybdate (ATTM) to re-lease H2 S and its cytoprotective effect on an oxidative injury model .METHODS:Released H2 S was absorbed in a reaction flask from ATTM dissolved in the cell medium .Staining with dichlorodihydrofluorescein diacetate or rhodamine 123 fol-lowed by photofluorography was conducted for the observation of reactive oxygen species ( ROS) and mitochondrial mem-brane potential (ΔΨm) levels, respectively.Cell viability and release of lactate dehydrogenase (LDH) from the cells were measured with commercial kits.RESULTS:Similar to another H2S donor GYY4137, ATTM had an ability to release H2S in the cell medium in a dose-dependent manner .Treatment of human skin HaCaT cells with ATTM at concentrations of 25~400 μmol/L didn’ t significantly alter cell viability .Exposure of the cells to ultraviolet rays or a ROS donor H 2 O2 in-creased the intracellular ROS levels .Treatment with 400 μmol/L H2 O2 significantly reduced the viability of HaCaT cells (P<0.01).However, before the treatment with H2O2, pretreatment with ATTM at 100 and 200 μmol/L markedly pre-vented the H2O2-induced cell injury (P<0.01).In addition, the treatment with H2O2 triggeredΔΨm loss (P<0.01) and LDH release from the cells (P<0.01).Prior to suffering from H2O2 injury, the preconditioning with 200 μmol/L ATTM significantly improved ΔΨm levels ( P<0.05 ) and attenuated LDH release from the cells ( P<0.01 ) .CONCLUSION:ATTM is capable of releasing H 2 S and protecting human skin cells against oxidative injury .

OBJECTIVE To study the protective effects of twin drugs of tetramethylpyrazine-scutellarein （TMSC4） on cerebral ischemia-reperfusion injury （CIRI） model rats and its mechanism. METHODS One hundred and five SD rats were randomly divided into sham operation group， model group， scutellarein group （0.7 mmol/kg）， tetramethylpyrazine group （0.7 mmol/kg）， and TMSC4 low-dose， medium-dose and high-dose groups （0.35， 0.7， 1.4 mmol/kg）， with 15 rats in each group. Sham operation group and model group were given constant volume of normal saline intragastrically， and other groups were given relevant drug intragastrically， once a day， for consecutive 14 d. Except for sham operation group， all other groups were treated to establish the CIRI model using the thread occlusion method. After 2 hours of ischemia and 22 hours of reperfusion， the brain index and brain water content of the rats were measured. Serum levels of interleukin 1β （IL-1β）， IL-6 and tumor necrosis factor α （TNF-α）， the levels of superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione peroxidase （GSH-Px） and catalase （CAT） in brain tissues， the situation of neuronal cell apoptosis， and the protein expressions of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax） and cleaved-caspase-3 were evaluated. RESULTS Compared with sham operation group， the brain index， brain water content， the serum levels of IL-1β， IL-6 and TNF-α， the levels of MDA in brain tissues， the brain cell apoptosis and the protein expressions of Bax and cleaved-caspase-3 in model group were significantly increased （P＜0.05）； the levels of SOD， GSH- Px and CAT and the protein expression of Bcl-2 in brain tissues were significantly decreased （P＜0.05）. Compared with model group， the above indexes of rats were reversed significantly in administration groups （P＜0.05）， while the reverse effects of TMSC4 medium-dose and high-dose groups were significantly better than those of scutellarein group and tetramethylpyrazine group （P＜0.05）. CONCLUSIONS TMSC4 has a certain protective effect in CIRI model rats， the mechanism of which may be related to relieving inflammatory reaction and oxidative stress， inhibiting cell apoptosis.