1.NOK Interacts With Akt and Enhances Its Activation
Yinghua LI ; Zhijie CHANG ; Li LIU
Progress in Biochemistry and Biophysics 2008;35(1):29-34
NOK is a newly identified receptor protein-tyrosine kinase (PRTK) molecule that can promote tumorigenesis and metastasis. Previous data showed that NOK could activate the phosphatidylinositol 3'-kinase (PI3K) pathway in stable BaF3 cells. But how does NOK activate PI3K in cells remains unknown. It was showed that NOK physically interacted with the PI3K downstream effector Akt and enhanced its activation in human embryo kidney 293T (HEK293T) cells. Deletion mapping indicated that protein kinase B (Akt) was able to directly contact with the kinase domain of NOK. Inactivating the Akt kinase domain significantly reduced the intermolecular interaction between NOK and Akt, while constitutively active mutant of Akt apparently had a stronger interaction with NOK. NOK did not have an additive effect on insulin-mediated Akt activation. Overall, the results indicate that NOK might complex with Akt and directly activate PI3K/Akt signaling pathway.
2.NOK Activates STAT3 Signaling by a JAK2-Dependent Mechanism
Yinghua LI ; Yu RONG ; Zhijie CHANG ; Li LIU
Progress in Biochemistry and Biophysics 2008;35(2):143-150
Novel oneogene with kinase-domain (NOK) can activate multiple mitogenic signaling pathways including the janus kinases(JAK) and signal transducer and activator of transcription proteins (STAT). It was showed that NOK specifically and physicallyinteracted with STAT3 in human embryo kidney 293T (HEK293T) cells. In addition, NOK could directly interact with most of the STAT3 subdomains except coiled-coil and C-terminal domains. Removing ectodomain and transmembrane domain of NOK markedly enhanced its intermolecular interaction with STAT3. Also, NOK could co-immtmoprecipitate with JAK2. in vivo. Importantly, co-expression of NOK and JAK2 produced a synergistic effect on NOK-mediated STAT3 activation, while inactivating the kinase domain of JAK2 completely prevented this synergistic effect. Overall, the results indicated that NOK might complex with both STAT3 and JAK2 and activate STAT3 signaling by a JAK2-dependent mechanism.
3.The cloning and expression of human interleukin-17 receptor-like molecule
Yiling HUANG ; Liming HUANG ; Shiqin XIONG ; Zhijie CHANG
Journal of Xi'an Jiaotong University(Medical Sciences) 1981;0(03):-
Objective To generate an human interleukin-17 receptor-like molecule (IL-17RLM) recombinant plasmid with 6?myc tag and detect its specific expression in eukaryotic cells. Methods Design two specific primers(including the enzyme sites of EcoRⅠand XhoⅠ), reextract hIL-17RLM-L DNA fragment after PCR and insert it into the 6?myc tagged pcDNA3.0 vector, then detect its expression by Western blot after transfecting COS7 cells. Results The 6?myc tagged recombinant plasmid pcDNA3.0- 6?myc /hIL-17RLM-L was generated successfully and its expression can be detected by Western blot in eukaryotic cells. Conclusion The eukaryotic expressing plasmid pcDNA3.0-6?myc /hIL-17RLM-L was generated successfully and its specific expression was realized, which may provide the basis for further research of its biological function.
4.Effect of cisplatin on ototoxicity and expression of caspase-3 in mouse cochlea
Weixia SUN ; Aimei WANG ; Cuifen BAO ; Lijie HUI ; Zhijie CHANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2004;0(05):-
Objective To establish a mice model of cisplatin-induced ototoxicity and to investigate the effect of cisplatin on the expression of caspase-3 in mouse cochlea.Methods Totally 69 Kunming mice were randomly divided into control group,cisplatin 2.5mg/(kg?d) group,cisplatin 3.5mg/(kg?d) group and cisplatin 4.5mg/(kg?d)group.Mice were injected intraperitoneally for 5 days.Auditory brainstem response(ABR) was measured to observe the change of hearing.Envision method of immunohistochemistry was applied to detect the expression of caspase-3 in cochlea.Results The weight and hearing of mice in different dose cisplatin groups were declined significantly as compared with those in control group(P
5.A LIF Mutation at The 29th Amino Acid Totally Abolished The Biological Functions
Hongxiu NING ; Yue CHEN ; Yuanjiang ZHANG ; Yu RONG ; Xiaojun WU ; Xiufang ZHANG ; Zhijie CHANG
Progress in Biochemistry and Biophysics 2005;32(4):318-324
Leukemia inhibitory factor (LIF) plays important roles in varieties of biological processes. This factor is highly conserved in mammalian animals and only one heterozygous LIF mutation was reported to cause the infertility of women. A LIF mutation was generated and the evidences were provided that the mutation of mature LIF at the 29th amino acid totally abolished its functions, including stimulation of STAT activation assayed by Luciferase reporter gene expression and EMSA experiments. In addition, the mutated LIF failed to inhibit the proliferation of M1 cells. The data indicated that the mutation of LIF did not have a dominant negative effect but lost the biological functions, suggesting that the 29th amino acid is critical for maintaining the activities of LIF.
6.effect ofTripterygium Wilfordii Hook. F. andTripterygium Hypoglaucum(Lévl.) Hutch on macrophage inflammatory factor
Chang XU ; Qingguo ZHAO ; Xiaohe XIAO ; Kuijun ZHAO ; Jiabo WANG ; Zhijie MA
International Journal of Traditional Chinese Medicine 2016;38(9):821-825
Objective To observe the effect ofTripterygium Wilfordii Hook. F. andTripterygium Hypoglaucum (Lévl.) Hutch on macrophage inflammatory factor, and to provide the oretical basis and experimental basis for the clinical application of these drugs.Methods Two batches ofTripterygium Wilfordii Hook. F. andTripterygium Hypoglaucum (Lévl.) Hutch were collected, and then the samples turned into alcohol extract by extraction and isolation. The IC50values of alcohol extracts were measured by MTT in BMDM cell. BMDM cell induced by the 4 batches of samples with IC50, then IL-6, IL-10, iNOS were detected by Elisa. Results The content of IL-6 (5.08 ± 0.96 pg/ml, 6.24 ± 0.20 pg/mlvs. 7.92 ± 0.84 pg/ml) and iNOS (0.14 ± 0.04 ng/ml, 0.36 ± 0.11 ng/mlvs. 0.86 ± 0.13 ng/ml) in Anhui and Guizhou groups were significantly lower than sulfasalazine (P<0.05), and the content of IL-10 (21.20 ± 4.24 pg/ml, 26.49 ± 4.44 pg/mlvs. 9.06 ± 0.40 pg/ml) in Anhui and Guizhou groups were significantly higher than sulfasalazine (P<0.05). The content of IL-6 (4.22 ± 0.38 pg/ml, 4.55 ± 0.44 pg/mlvs. 7.92 ± 0.84 pg/ml) and iNOS (0.07 ± 0.04 ng/ml, 0.28 ± 0.10 ng/mlvs. 0.86 ± 0.13 ng/ml) in Hunan and Zhejiang groups were significantly lower than sulfasalazine (P<0.05) .Conclusion The anti-inflammatory effect ofTripterygium WilfordiiHook. F. treat rheumatoid arthritis is better than sulfasalazine andTripterygium Hypoglaucum (Lévl.) Hutch.
7.Efficacy of Berberine Hydrochloride Tablet in the Treatment of Non-alcoholic Fatty Liver Disease:A Me-ta-analysis
Chang XU ; Xuelong LIU ; Jianteng NI ; Zhen WU ; Zhijie MA ; Kuijun ZHAO
China Pharmacy 2015;(30):4232-4235
OBJECTIVE:To systematically review the efficacy of Berberine hydrochloride tablet in the treatment of non-alco-holic fatty liver disease(NAFLD),and provide evidence-based reference for the clinical treatment. METHODS:Retrieved from CJFD,Wanfang Database,VIP,CBM and PubMed,observational studies about Berberine hydrochloride tablet in the treatment of NAFLD were collected. After data extraction and quality evaluation,Meta-analysis was performed by using Rev Man 5.3 statistics software. RESULTS:A total of 6 studies were included,involving 294 patients. Results of Meta-analysis showed Berberine hydro-chloride tablet could significantly reduce the levels of AST[WMD=18.97,95%CI(2.25,35.70),P=0.03],ALT[WMD=31.04, 95%CI(7.17,54.91),P=0.01],TG[WMD=1.07,95%CI(0.39,1.74),P=0.002] and TC[WMD=1.31,95%CI(0.79,1.84),P<0.001] in the serum of patients with NAFLD. There were significant differences. CONCLUSIONS:Berberine hydrochloride tablet can significantly improve the liver function and blood lipid levels of patients with NAFLD,and the clinical efficacy is relatively pre-cise. Due to the limit of methodological quality,it remains to be further verified by large-scale and high quality RCT.
8.The effect of different regions of Tripterygium Hypoglaucum (Lévl.) Hutch on macrophage inflammatory factor
Chang XU ; Qingguo ZHAO ; Xiaohe XIAO ; Kuijun ZHAO ; Jiabo WANG ; Zhijie MA
International Journal of Traditional Chinese Medicine 2015;(11):1005-1009
Objective To observe the effect of different regions of tripterygium hypoglaucum (Lévl.) hutch on macrophage inflammatory factor, and to providetheoretical basis and experimental basis for the clinical application of tripterygium hypoglaucum (Lévl.) hutch. Methods 5 batches of tripterygium hypoglaucum (Lévl.) hutch were collected, then the samples turned into alcohol extract by extraction and isolation. The IC50 values of alcohol extracts were measured by MTT in BMDM cell. B MDM cell were induced by the 5 batches of samples with IC50, then IL-6, IL-10, iNOS were detected by Elisa. The efficacy of different regions of tripterygium hypoglaucum (Lévl.) hutch on macrophage inflammatory factor was evaluated by comparison with sulfasalazine. Results The content of IL-6 (4.22 ± 0.38 pg/ml, 4.55 ± 0.44 pg/ml vs.7.92 ± 0.84 pg/ml) and iNOS (0.07 ± 0.04 ng/ml, 0.28 ± 0.10 ng/ml vs. 0.86 ± 0.13 ng/ml) in HuNan and ZheJiang groups were significantly lower than sulfasalazine (P<0.05), and the content of IL-10 (19.34 ± 6.06 pg/ml, 24.34 ± 3.03 pg/ml vs. 9.06 ± 0.40 pg/ml) in Guizhou and Fujian groups were higher than sulfasalazine (P<0.05). Conclusion The anti-inflammatory effect of HuNan and ZheJiang's tripterygium hypoglaucum (Lévl.) hutch treat rheumatoid arthritis is better than sulfasalazine, so theyaregenuine regional drug in the treatment of rheumatoid arthritis. Additional research will analyze associations between tripterygium hypoglaucum (Lévl.) hutch and rheumatoid arthritis.
9.Protective effect and mechanism of ulinastatin on rats with hemorrhagic shock
Longyuan JIANG ; Lianhong YANG ; Wa ZHONG ; Zhijie HE ; Jianxing CHANG ; Tao YU ; Tong WANG ; Zitong HUANG
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To investigate the protective effect of ulinastatin on rats with hemorrhagic shock. METHODS: A prospective, controlled animal study was designed. The model of hemorrhagic shock in rats was produced by Chaudry method. After 60 min, rats were resuscitated by transfusion of shed blood and normal saline, but a half of them were treated with ulinastatin. At different time points after reperfusion, the levels of tumor necrosis factor-alpha (TNF-?), interleukin-6 (IL-6), malondialdehyde (MDA) and superoxide dismutase (SOD) in serum were detected. RESULTS: The levels of TNF-?, IL-6 and MDA significantly increased and the activity of SOD decreased. In the ulinastatin-treated groups, the blood pressure and heart rate were obviously improved; the levels of TNF-?, IL-6 and MDA significantly decreased and the activity of SOD had little change after hemorrhagic shock and reperfusion. CONCLUSION: Ulinastatin has a protection effect on rats with hemorrhagic shock by suppressing the production of inflammatory factors and reducing oxidative damage.
10.c-Cbl Mediated Ubiquitination and Degradation of hSef
Yongming REN ; Zhili RONG ; Zhiyong LI ; Long CHENG ; Yinghua LI ; Yinyin WANG ; Fangli REN ; M.irwin DAVID ; Zhijie CHANG
Progress in Biochemistry and Biophysics 2008;35(1):43-49
Sef (similar expression to fgf genes) was identified as a feedback antagonist of FGF signaling in zerbrafish, mouse and human. Sefhas been reported to function in different ways, however the regulation of Sef stability remains unknown. The possible role of c-Cbl in the regulation of Sef protein degradation was investigated. Results from coimmunoprecipitation and immunostaining assays reveal that hSef colocalizes and interacts with c-Cbl. Data suggest that the interaction between hSef and c-Cbl results in the ubiquitination and subsequent degradation of the hSef protein. It was proposed that c-Cbl may serve as a modulator to regulate Sef protein stability during FGF signal transduction.