1.IL-1β promotes glial scar formation after spinal cord injury in rats by JAK2-STAT3
Jingxian LIU ; Yongzhi XIA ; Fugui WANG ; Wei TANG ; Yi YAN
Basic & Clinical Medicine 2017;37(5):668-675
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.
2.Inhibitory effect of Adp53 and F56 on growth and metastasis of transplantation of the breast tumor in mice
Shaowen XIAO ; Bo YAN ; Jingxian YANG ; Chengchao SHOU ; Shanwen ZHANG
Journal of Peking University(Health Sciences) 2004;0(02):-
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.
3.Influence of all-trans retinoic acid on the methylation of the RARa gene in ovarian cancer cell line COC2
Yan JU ; Qi LIU ; Yuanzhe WU ; Jingxian LIN
Journal of Medical Postgraduates 2003;0(05):-
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.
4.Osthole reduced Aβ synthesis by up-regulatingmiRNA-107 in neurons transfected with APP595/596 gene
Honghe XIAO ; Yanan JIAO ; Yuhui YAN ; Hongyan LI ; Jingxian YANG
Chinese Pharmacological Bulletin 2017;33(8):1079-1085
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.
5.Clinical application of preoprative TAE in the nasopharyngeal angiofibromas
Yue LIU ; Zhiping YAN ; Jingxian ZHANG ; Wenheng TANG
Journal of Interventional Radiology 2001;0(06):-
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)
6.In vitro cytotoxicity test of medical ultrasonic couplant.
Jingxian GAO ; Shasha WANG ; Meng JIN ; Xiaoli YAN
Chinese Journal of Medical Instrumentation 2013;37(3):210-212
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.
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Colorimetry
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Cytotoxicity Tests, Immunologic
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7.Neurotrophin 3 gene overexpression promotes cholinergic differentiation in neural stem cells
Yuhui YAN ; Shaoheng LI ; Liang KONG ; Yingjia YAO ; Yanan JIAO ; Zhenyu TAO ; Jie SONG ; Jingxian YANG
Chinese Pharmacological Bulletin 2016;32(5):631-637
Aim To investigate the effects of neurotro-phin-3 ( NT-3 ) gene overexpression on the differentia-tion into cholinergic neuron of neural stem cells ( NSCs) in vitro and its underlying mechanism. Meth-ods Brain-derived NSCs from newborn mice were iso-lated and cultured in vitro and determined by immuno-fluorescence. The NSCs were divided into three groups: NSCs, GFP-NSCs and NT-3-NSCs groups. The expression of NT-3 was detected by immunofluo-rescence and ELISA. Then, the ability of NSCs on dif-ferentiation into cholinergic neuron was detected by im-munofluorescence and RT-PCR, and the Acetylcholine Assay Kit was used for acetylcholine ( ACh) , and the expression of Hes1 , Mash1 and Ngn1 mRNA was de-termined by RT-PCR. Results The neurosphere dis-played Nestin and Sox 2-postive by immunofluores-cence, suggesting that the cultured cells were NSCs. The proportion of ChAT immunopositive cells was sig-nificantly higher in the NT-3-NSCs group than that in the other two groups ( P <0. 01 ) . Ach secretion in NT-3-NSCs was significantly elevated compared with the other two groups ( P <0. 01 ) . NSCs transfected with NT-3 increased the levels of Mash1 and Ngn1 mR-NA, and decreased the level of Hes1 mRNA ( P <0. 05 ) . Conclusion NT-3 can significantly promote the in vitro differentiation of NSCs into cholinergic neu-rons via probablly inhibiting Notch signaling pathway.
8.Protective effect of osthole on SH-SY5Y cells transfected with APP595/596 gene
Yanan JIAO ; Yingjia YAO ; Liang KONG ; Shaoheng LI ; Zhenyu TAO ; Yuhui YAN ; Jingxian YANG
Chinese Journal of Pathophysiology 2015;(11):2053-2058
AIM:To explore the protective effect of osthole on the SH-SY5Y cells transfected with APP595/596 gene, and to investigate the molecular mechanism.METHODS:The SH-SY5Y cells were transfected with APP595/596 gene in vitro for establishing a cell model to study the pathogenic role of amyloid β-protein ( Aβ) .The cell viability was detected by CCK-8 assay.The release of lactate dehydrogenase ( LDH) was determined by the colour reaction of dia-phorase-INT.The cell apoptotic rate was analyzed by flow cytometry.The expression of β-site APP cleaving enzyme 1 ( BACE1) at mRNA and protein levels was detected by RT-PCR and Western blot.The expression of Aβwas measured by the technique of immunofluorescence cytochemistry and Western blot.RESULTS: Treatment with osthole inhibited the LDH release, and increased the viability of the cells.The percentage of apoptotic cells was also significantly decreased. Osthole also inhibited the expression of BACE1 at mRNA and protein levels and the protein expression of Aβ.CONCLU-SION:Osthole has protective effect on SH-SY5Y cells transfected with APP595/596 gene.The mechanism may be associ-ation with inhibiting the mRNA and protein expression of BACE1.
9.Acupuncture Effects on Endocranial Hemodynamics of Patients with Takayasus Arteritis(Brachiocephalic Artery Type)
Guangxia NI ; Jingxian HAN ; Qifang GAO ; Li YAN ; Zhi ZHAO ; Xuemin SHI
Journal of Acupuncture and Tuina Science 2007;5(6):362-365
Objective: To discuss the action mechanisms of Takayasus arteritis (brachiocephalic artery type) of acupuncture. Methods: We applied acupuncture therapy with the principle of warming Yang and supplementing Qi, removing obstruction in the meridians and recovering pulse, to treat patients with takayasus arteritis (brachiocephalic artery type). Before and after treatment, we detected the TCD (Transcranial Doppler) changes of the average velocity of blood flow (AVBF) and arterial pulsatility index (PI) of related arteris. Results: After treatment,acupuncture and moxibustion can regulate the abnormal blood flow rate in endocranial correlated arteries by two sides, in the AVBF while, elevate the elasticity of endocranial vessels.Conclusion: Acupuncture and moxibustion can regulate cerebrovascular function of patients with takayasus arteritis (brachiocephalic artery type), increase the perfusion of cerebral blood flow, and adjust abnormal state of endocranial hemodynamics. This is possibly the important action mechanisms acupuncture in treating this disorder.
10.Osthole promotes differentiation into neurons and reduces neuronal apoptosis via Wnt/β-catenin signaling pathway in APP transduced neural stem cells
Yingjia YAO ; Liang KONG ; Yanan JIAO ; Shaoheng LI ; Zhenyu TAO ; Yuhui YAN ; Jingxian YANG
Chinese Pharmacological Bulletin 2015;(11):1516-1523
Aim To investigate the effects of osthole ( Ost) on the ability of proliferation and differentiation in APP transduced neural stem cells( NSCs) , and neu-ronal apoptosis, in order to find related mechanism. Methods A model of Alzheimer′s disease( AD) cells was successfully established by transducing APP gene into NSCs in vitro. The ability of proliferation and dif-ferentiation was tested by staining. The viability of NSCs was determined by using CCK-8 assay. The cell apoptosis was tested by Hoechst 33258 staining. The expression of GSK-3β and β-catenin mRNA was deter-mined by RT-PCR. The expression of GSK-3β and β-catenin protein was determined by Western blot. Re-sults The ability of proliferation had increased by 10 . 24% with Ost treatment, compared with APP group. The ability of differentiation had increased by 6 . 74%with Ost treatment, compared with APP group. The vi-ability of NSCs had increased and cell apoptotic rate had decreased significantly. From the results of RT-PCR and Western blot, we could find the expression of GSK-3βmRNA and protein had decreased, and the ex-pression of β-catenin mRNA and protein had increased significantly, compared with APP group. Conclusion Ost could enhance the ability of proliferation and dif-ferentiation into more neurons of NSCs transducing APP gene, and reduce neuronal apoptosis. It might be relat-ed with activiting Wnt/β-catenin signaling pathway.