Objective To investigate the regulation on cell cycle related factor such as Cyclins and P21waf/cip1 by inhibiting histone deacetylase(HDAC)with valproic acid sodium(VPA).Methods HepG2 hep-tocellular carcinoma cells.BGC-823 gastric carcinoma cells and MCF-7 breast cancer cells were cultured with O.75-4.00 mmoL/L VPA for 48 h in vivo.and the cell cycle was analyzed by flow cytometry with PI assay.The protein and mRNA expression of Cyclin A,Cyclin D1,Cyclin E and P21waf/cip1 were analyzed by indirect immu-nofluorescence technique and RT-PCR.respectively.Results Compared with control groups,VPA at concen-trations 0.75-4.00 mmol/L exerted a significant inhibiting effect on G1 phase of HepG2,BGC-823 and MCF-7 cells(P<0.001).and the effect was dose dependent.Cyclin A was down-regulated both at mRNA and protein level in HepG2 and BGC-823 cells(P<0.001),but no difference in MCF-7 cells(P>0.05).Cyclin D1 was down-regulated both at mRNA and protein level(P<0.001)and P2lwaf/cip1 was up-regulated both at the mRNA and protein level in the three cell lines(P<0.001);Conversely,protein and mRNA expression of Cyclin E were unchanged upon treatment with VPA(P>0.05).Condusion Acetylization of histone intervened with VPA can regulate Cyclin D1 and P21waf/cip1 expressions obviously.To the expression of Cyclin A,it shows some difference according to the histogenesis and phenotypes of different carcinoma types.But there is not any obvious function on Cyclin E.Down-regulating Cychn D1 and up-regulating P21waf/cip1 may be the common target path-way in the inhibition of cell cycle G1 phase exerted by VPA.

Background Corneal transplantation is a primary method for the treatment of serious corneal diseases, but its application is limited because of the shortage of corneal donor.The study on tissue engineering corneal epithelium provides a new approach to corneal transplantation, and the biological scaffold materials for tissue engineering corneal epithelium is an issue of increasing concern.Bacterial cellulose membrane has been used in medical field,but its application in tissue engineering corneal epithelium deserves more researching.Objective This study was to evaluate the biocompatibility of bacterial cellulose membrane as a biological scaffold of tissue engineering corneal epithelium.Methods Corneal epithelium was isolated from 1 month-old New Zealand White rabbit.Corneal epithelial cells were cultured using explant method and identified by detecting the CK-3 expression using immunofluorescence technique.The second generation ceils were inoculated on bacterial cellulose membrane and culture plate, respectively, and the growth status of the cells were examined and compared under the optical microscope.The cell activity/toxicity test was performed by LIVE/DEAD cell staining kit at the third day after inoculation to evaluate the survival rate.The ultrastructure of the cell surface was examined under the scanning electron microscope.The study was performed in accordance with the ARVO Statement.Results Rabbit corneal epithelial cells grew well 1 week after primarily cultured with a cobblestone-like appearance and positive response for CK3 antibody.The cells on the bacterial cellulose membrane presented a round shape and regular arrangement and showed the green fluorescence for LIVE/DEAD test,with the survival rate 100％.Abundant leafy protrusion, microvilli and intercellular junction were seen under the scanning electron microscope.In addition, mitosis phase of cells and many filopodia between the cells and bacterial cellulose membrane were also exhibited.Conclusions Rabbit corneal epithelial cells can grow well in bacterial cellulose membrane.Bacterial cellulose membrane has good biocompatibility, indicating that bacterial cellulose membrane can be used as new biological material for tissue engineering corneal epithelium.

Chaihu Shugan Powder has the function of soothing liver and promoting qi flow, activating blood and relieving pain. In clinic, it is mainly used for the syndrome of liver qi stagnation, with wide application. This article reviewed the literature about effects of Chaihu Shugan Powder on neurobiochemistry, endocrine regulation, immunity and antioxidant pharmacological effects and mechanism, with a purpose to provide references for clinical medication and new medicine research.

Medical consumable is widely used in disease diagnosis,treatment and health care,but its multifarious classifications show its weakness in management.Based on the research and analysis on the existing kinds of classifications,this article unifies practical application in hospital and proposes"null-stock"classification method under the management plan,which seems more scientific and reasonable.

Objective To investigate the effect and the mechanism of up-regulating histone acetylizad level with a selective inhibitor of HDACs-Valproate acid sodium (VPA) on breast cancer cell proliferation. Methods MCF-7 cells were cultured with 0.75-4.0 mmol/L valproic acid (VPA) for 24, 48, 72, 96 hours in vitro, the inhibiting rate was tested by MTT assay. Cell cycle was analyzed by flow eytome- try with PI assay, and the protein and mRNA expressions of Cyelin A, Cyclin DI, Cyclin E, P21Waf/cipl of MCF-7 cells after 1.5, 3.0 mmol/ L VPA treated were analyzed by indirect immunofluorescence technique and RT-PCR respectively. Results After cultured with 0.75 -4.0 mmol/L valproic acid (VPA) for 24, 48, 72, 96 hours, the inhibiting rate of experimental groups increased significantly(P<0.01) and a dose and acting time dependent manner was found. As to cell cycle, the percentages of GI, S, M phrase in control groups remained the same. Contrary to control groups, 0. 75 -4.0 mmo]/L VPA induced a significant arrest in G1 phrase ( P<0.01), and a total of 55.4% -82.8% G1 phrase ratio were found. P21Waf/cipl was up-regulated both at the mRNA and protein level while Cyclin D1 was down-regulated ( P<0.001). Conversely, neither mRNA nor protein expression of Cyclin A, Cyclin E showed difference ( P>0.05). Conclusions Up- regulating histone acetylizad level can inhibit breast cancer cell proliferation, induce cell cycle arrest in G1 phrase. VPA, as a I class of histone deaeetylase inhibitor, can be used as an option in the treatment of breast cancer. The mechanism may include up-regulating P21Waf/cipl mRNA and protein expression and down-regulating Cyclin D1 mRNA and protein expression.

AIM: In order to explore the mechanism of the GC - like effect of Tsiao Shaihutang (TSS), the down - regulation of glucocorticoid receptor and mRNA by TSS were studied. METHODS: GR sites were determined by receptor radio ligand binding assay method. At the same time, GR mRNA level was determined by quantitative reverse- transcriptive polymerase chain reaction (RT- PCR) method. RESULTS: (1) GR sites and GR mRNA level were down - regulated significantly after GC, TSS (P < 0.01 ) treatment . (2) GR sites and GR mRNA level in GC plus TSS group were obviously higher than those in GC group (P < 0.01 ). CONCLUSIONS: These data suggested that TSS can significantly down- regulate GR and GR mRNA level.

Objective To study the effects of bone marrow mesenchymal stem cells modified by brain-derived neuro-trophic factor ( BDNF) gene on the repair of spinal cord injury by electrophysiological assay .Methods Thirty healthy Spra-gue-Dawley rats (male and female) were randomly divided into 3 groups:Blank group, 10 rats (removal of the lamina only and exposed spinal dura mater );spinal cord injury (SCI) group,10 rats;and cell transplantation after SCI group , 10 rats. Eight rats of them were selected randomly and detected their SEP and MEP , and evaluated the degree of recovery of motor scores in the rats at 1 d, 7 d, 14 d, 21 d, 30 d, and 60 d.Result Since 4 days after cell transplantation , the process of hind limbs changes was as follows:at the 1-4 days after injury , the injury side hind limb had flaccid paralysis , mopping the floor walk, the movement of contralateral hind limb was gradually recovered from the initial injury , the injury side hind limb had spastic paralysis in 5-9 days after SCI;during 10-14 days, the injury side had a few activities;the contralateral side re-covered to a less normal state;At 15-21 days, activities of the injury side improved obviously , until the 30th day.The activ-ity and muscle tension degree of the injury side recovered most obviously .After 30 days no more obvious improvement was ob-served.Immunohistochemistry showed that the transplanted mesenchymal stem cells , which were induced and labeled firstly , survived at the damage spinal cord , and behavioral observation found that the cell transplantation improved exercise capacity of the rats injured before .Conclusion Bone marrow mesenchymal stem cells modified by BDNF gene can partially promote the recovery of nerve transmission function and nerve regeneration .

Objective To explore the effect of Chaihu Shugan Powder (CSP) on the content of 5-hydroxytryptamine (5-HT) in the hippocampus and the expression of tryptophan hydroxylase 2(TPH2) in median raphe nuclei in depression rats.Methods SD rats were randomly divided into normal group(n=12),model group (n=12),fluoxetine group (n=12),low-dose and high-dose CSP group (n =12,respectively).Depression model was made by reserpine intraperitoneal injection.During the experiment,the weight and the open-field scores were calculated;the content of 5-HT was detected by ELISA.The expression of TPH2 in median raphe nuclei was detected by Western blot.Results Compared with the weight ((225.02±5.23) g),the open-field scores ((12.6± 5.1)score) and content of 5-HT ((1.09±0.27) ng/ml) in the model group,high-dose CSP showed significantly improve the depressive rats in weight,open field score and content of 5-HT ((238.78±5.16) g,(15.6±7.8) score and (1.80±0.58) ng/ml,respectively;P＜0.05 or P＜0.01).The expression of TPH2 (0.66±0.21) in median raphe nuclei in the high-dose CSP group was apparently increased compared with that in the model group(0.16±0.04) (P ＜0.01).Conclusion CSP have the effects of anti-depression,which could be related with the increase of the 5-HT content in the hippocampus and the expression of TPH2 in median raphe nuclei.

Objective:To investigate modulation of a specific HDAC inhibitor,Valproate acid sodium(VPA),on expression of Caspase3,Caspase8,Caspase9 by inhiting HDAC,as well as apoptosis rate of cancer cells treated with VPA and the specific inhibitors of Caspase3,Caspase8,Caspase9.Methods:Heptocellular carcinoma cells-HepG2,gastric carcinoma cells-BGC-823 and breast cancer cells-MCF-7 were cultured with 0.75-4.0 mmol/L of VPA for 48 hrs in vivo,apoptosis was analyzed by flow cytometry with Annexin V/PI assay.The activities and protein expressions of Caspase3,Caspase8,Caspase9 were detected by spectrophotometry and indirect immunofluorescence technique.Results:Contrary to control groups,VPA at concentrations between 0.75 and 4.0 mmol/L induced a significant apoptosis in HepG2,BGC-823,MCF-7 cells(P0.05).The apoptosis rates of cancer cells treated with VPA and specific inhibitors of Caspase3,Caspase9 together was lower than in the groups with VPA treatment singlely(P

The traumatic brain injury(TBI)is the main cause of death and disability after earthquake.The rehabilitation should intervene as earlier as possible.This paper would introduce some rehabilitation approaches practically.