Aim To investigate the neuroprotective effects of osthole(Ost)on the primary cultured cortical neurons transfected with APP595/596 gene and its underlying mechanism.Methods Neonatal mouse cortical neurons were transfected with APP595/596 gene to establish AD cell models for the further study.Then,the cell viability was detected by CCK-8 assay,and the leakage of lactate dehydrogenase(LDH)was assayed by LDH kit to evaluate the injury degree.Transferase-mediated nick end labeling(TUNEL)was used to evaluate the cell apoptosis.The expression of β-amyloid peptide(Aβ)and β-site APP cleaving enzyme 1(BACE1)was measured by immunofluorescence,while the miRNA-107 was measured by RT-PCR.Results Compared to model group,Ost could significantly improve the neurons viability,decrease the LDH release and prevent the apoptosis.Ost also inhibited the expression of Aβ and BACE1 at protein level,while enhanced the expression of miRNA-107 at gene level.Conclusion Ost plays a neuroprotective role in neurons transfected with APP595/596 gene in part through up-regulating miRNA-107.

Objective To investigate the mechanism of IL-1β in promoting glial scar formation after spinal cord injury.Methods The experimental model of SCI was created by extradural compression of the spinal cord using an aneurysm clip.Rats were randomly divided into model group, sham operation group, IL-1β inhibitor IL-1RA group, IL-1β group and IL-1β+JAK2-STAT3 inhibitor AG490 group, according to different interventions, then were given normal saline, IL-1RA, IL-1β and IL-1β+AG490 every 10 μL respectively, sham group received only laminectomy.The motion function of the hindlimbs of rats was measured by Basso Beattie Bresnahan(BBB) scores and the expression of GFAP, vimentin and p-STAT3 were detected by Western blot technique, immunofluorescence assay and immunohistochemistry technique at corresponding time points(at the 8th, 12th hour, 1st, 3rd, 7th and 14th day after SCI).Results The expression trend of p-STAT3(at the 8th and 12th hour after SCI),GFAP and vimentin(at the 7th and 14th day after SCI)was: the expressions of p-STAT3, GFAP and vimentin in the model group were significantly higher compared with the sham group(P<0.01), the expression of p-STAT3,GFAP andvimentin in the IL-1RA group were significantly lower compared with the model group(P<0.05) whereas significantly higher compared with the sham group(P<0.05);the expressions of p-STAT3, GFAP and vimentin in the IL-1β+AG490 group were significantly lower compared with the model group(P<0.05)whereas significantly higher compared with the sham group(P<0.05), the expressions of p-STAT3, GFAP and vimentin in the IL-1β group were significantly higher compared with the model group(P<0.05).Conclusions IL-1β can improve glial scar formation via JAK2-STAT3 signal.Inhibition of IL-1β or JAK2-STAT3 can reduce glial scar formation and promote functional recovery of spinal nerve.

Objective:To investigate the effects of Adp53 and F56 on the growth and lung metastasis of breast cancer.Methods:The BICR-H1 cells were inoculated into the mammary fatty pad of BALB/C nude mice and NOD/SCID mice to establish breast cancer model.Then the nude mice with xenograft tumor were randomized into group Adp53+F56,Adp53,F56 and control.The NOD/SCID mice with xenograft tumor were randomized into group Adp53+F56,Adp53,F56,Adlacz and control.They were theated for 3 weeks according to the plan,diversity of the volume and histopathology of xenograft tumor of nude mice was observed and the expressions of p53 and VEGF gene,and microvessel density(MVD)were detected by immunohistochemistry.Lung metastasis of breast cancer in NOD/SCID mice was observed.Results:(1)Intratumoral injections of Adp53,F56,and their combination resulted in an inhibition on the growth of xenograft tumor of BICR-H1 cells.The ultimate relative growth volumes of groups Adp53+F56,Adp53,F56 and control were 2.47,4.37,4.69 and 12.49 respectively.(2)After treatment,P53 positive rate of group Adp53+F56,Adp53 increased 9.4%,6.3% than before respectively,but compared with control group,the difference is not significant(P=0.693);VEGF protein of group Adp53+F56,Adp53 and F56 decreased 21.9%,9.4% and 3.1% than before respectively,but compared with control group,the difference was not significant(P=0.284).Necrosis and decrease of vessel in the tumor and morphological change of endothelium were observed under light microscope in the groups Adp53+F56,Adp53 and F56.MVD estimated by FⅧ-RA staining of group Adp53+F56,Adp53 and F56 were 14.50?2.54,16.28?3.44 and 18.06?7.66,compared with control group(24.93?6.53),the difference is significant(P=0.000).(3)The average number of lung metastasis of NOD/SCID mice in group Adp53+F56,Adp53 and F56 were 1.143?0.378,2.750?0.886 and 3.375?0.518 respectively,lower than Adlacz group(5.000?0.816)and control group(5.670?0.817)obviously(P=0.000).Conclusion:Adp53 combined with F56 can greatly inhibit growth and matastasis of breast cancer in vivo.The mechanism of anti-tumor effects of Adp53 and F56 may be related to the anti-angiogenesis effect on malignant tumor through inhibiting the expression and activity of VEGF.

Objective To evaluate the clinical value of the preoperative intra-arterial embolization of the nasopharyngeal angiofibromas. Methods The treatment group of 7 male patients with the nasopharyngeal angiofibromas were undergone angiographic evaluation and embolization of tumor-feeding vessels before surgery. All patients were embolized with gelfoam particles and PVA. The control group of 7 patients received surgical treatment without preoperative embolization. We compared the volumes of intraoperative bleeding and the blood transfusions during oprations between the two groups. Results All patients achieved symptomatic remission,with no complications. Comparing with the control group,the amount of intraoperative bleeding and the blood transfusions during operations were much less in the treatment group submitted to endovascular embolirzation.Marked edema in the peripheral region of tumor of the treatment group made the tumor easy to be dissociated. Conclusion The intraoperative bleeding can be reduced significantly by preoperative embolization of supplying arteries to the nasopharyngeal angiofibromas,therefore it should be used routinely as an adjunct to surgery. (J Intervent Radiol,2006,15: 345-347)

Objective: All-trans retinoic acid(ATRA) is a classic drug that can induce tumor differentiation,and its influence on the aberrant methylation of cancer cells has been studied insufficiently.The objective of this study was to observe the influence of ATRA on the methylation of the RARa gene promoter in ovarian cancer cell line COC2 and its relationship with the RARa expression.Methods: The ovarian cancer cell line COC2 was treated with different concentrations of ATRA for different times.Methylation specific PCR(MSP) and bisulfate sequencing methods were used to detect the changes in the methylation of the RARa promoter after ATRA treatment.RT-PCR was employed to observe the changes in the expression level of RARa mRNA.Results: Aberrant methylation of the RARa gene promoter was found in the ovarian cancer cell line COC2.ATRA treatment decreased the number of RARa promoter methylation sites in COC2 within a certain scope in a concentrationand time-dependent manner,and increased the expression of RARa mRNA.Conclusion: Aberrantly high methylation of the RARa promoter exists in the ovarian cancer cell line COC2;ATRA can partially reverse the methylation and increase the RARa mRNA expression.

The cytotoxicity of medical ultrasonic couplant was tested by MTT assay and agar overlay test. By MTT assay, when the inoculum density was high, the cytotoxicity level was low, or vice versa. The cytotoxicity grade tested by agar overlay was not accord to MTT assay's too. MTT assay is suitable to test the cytotoxicity of medical ultrasonic couplant because it is quantitative and more sensitive, however, the experimental condition and the preparative method of extraction should be determined.
Animals ; Cell Line ; Colorimetry ; Cytotoxicity Tests, Immunologic ; methods ; Mice ; Ultrasonics

Objective: To observe the influence of all-trans retinoic acid(ATRA) on the expressions of E-cadherin,heparanase and VEGF in epithelial ovarian carcinoma cell line COC2,and to investigate the anti-metastatic potential and possible action mechanism of ATRA.Methods: We used flow cytometry to examine the expressions of E-cadherin,heparanase and VEGF proteins in the epithelial ovarian carcinoma cell line COC2 treated with different concentrations of ATRA.Results: ATRA significantly increased the expression of E-cadherin and decrease that of VEGF and hepareanse in a dose-dependent manner.Conclusion: ATRA can inhibit cell proliferation,improve cell-cell adhesion and downregulate the expressions of VEGF and heparanse proteins,suggestive of an anti-angiogenic and anti-metastatic potential.

BACKGROUND:Studies have found that adipose-derived stem cells (ADSCs)/collagen complexes can promote the ADSCs differentiation and maturation into mature adipocytes and promote angiogenesis.OBJECTIVE:To explore the biological properties of the ADSCs/collagen sponge composite material and to detect its effect on angiogenesis.METHODS:(1) ADSCs were cultured on collagen sponge (experimental group) or cultured alone (control group).After 24 hours of culture,cell adhesive rate of ADSCs was determined with flow cytometry.After 2,4,6 days of culture,cell proliferation and level of vascular endothelial growth factor (VEGF) in the culture medium were detected.(2) Chick embryo chorioallantoic membranes were exposed and incubated for 7 days and then divided into four groups:0.2 mL of sterile PBS was added in the blank group,0.2 mL of 2× 108/L passage 3 ADSCs suspension was added in the ADSCs group,collagen sponge was added in the collagen sponge group,and collagen sponge with 0.2 mL of 2× 108/L passage 3 ADSCs suspension was added in the composite group.After 7 days of incubation,the microvessel count around the chorioallantoic membrane was measured.RESULTS AND CONCLUSION:(1) The cell adhesive rate of ADSCs to collagen sponge reached to (93.04±0.67)％.(2)The absorbance value (at 6 days of culture) and level of VEGF (at 4 and 6 days of culture) in the experimental group were significantly higher than those in the control group (P ＜ 0.01 or P ＜ 0.05).(3) Compared with the blank group,the number of microvessels was significantly higher in the ADSCs,collagen sponge and composite groups (P ＜ 0.05).Moreover,higher amount of microvessels were found in the composite group than the ADSCs and collagen sponge groups (P ＜ 0.05).To conclude,ADSCs can adhere well to the collagen sponge with good biocompatibility and their combined use can improve angiogenesis further by enhancing cell proliferation and VEGF secretion of ADSCs.

Aim To investigate the effects of osthol on cell apoptosis and inflammatory cell infiltration after brain stab wound injury in mice. Methods The mice underwent the stab wound injury by a needle, then were randomly divided into sham operation group, model group, osthol 10, 20, 30 mg · kg-1 treatment group. The main examinations included mice brain wa-ter content; the apoptotic cytokines Bax, Bcl-2, Caspase-3 mRNA expression were assessed by PT-PCR; immunohistochemistry staining was used to de-tect neutrophils (MPO) and microglia (Iba-1) infiltra-tion and Caspase-3 positive cell expression around in-jured lesions. Results Treatment with osthole 20, 30 mg·kg-1 group significantly reduced the water content in injured brain, improved the ratio of Bax/Bcl-2, and reduced the expression of apoptosis cytokine Caspase-3 mRNA. Osthole 30 mg·kg-1 treatment group obvious-ly reduced the infiltration of neutrophils and microglial cells and significantly reduced the number of apoptotic cells around the injured cerebral cortex. Conclusion Osthole has therapeutic effect on stab wound injury in mice, and the possible mechanism may be by reducing the infiltration of inflammatory cells and reducing apop-totic cells.

Aim To investigate the effect of osthole on neuron synapses infected APP gene and its underlying mechanism. Methods The neurons were divided into three groups:GFP, APP, APP+Ost groups. The neu-rons were infected APP gene with containing mutational site in vitro for mimicking the characterstics of Alzhei-mer’ s disease ( AD) . The cell viability was assessed by CCK-8 , the expression of synapsin-1 was deter-mined by immunofluorescence, and the concentration of PSD-95 and SYP were detected by ELISA. The ex-pressions of Aβ1-42 , CAMKK2 , phoshorylated AMPKα1 , AMPKα1 protein were determined by West-ern blot. Results Strong APP staining was visible in neurons infected with APP and abundant expression of Aβ1-42 , a neurotoxic oligomer. Compared with APP group, APP+Ost group significantly increased cell vi-ability, promoted the expression of synapsin-1, up-reg-ulated the concentration of PSD-95 and SYP, and de-creased the expressions of CAMKK2 and p-AMPKα1 . Conclusions Ost can protect the neuron synapses a-gainst infected with APP gene. Its neuroprotective effect may be related to inhibiting the CAMKK2/AMPK signal pathway.