1.Molecular mechanism of Andrographolide on MMP-9 expression in human lung cancer cells
Xiangyu LUO ; Weimin LUO ; Xuesong ZENG
Chinese Journal of Biochemical Pharmaceutics 2014;(2):13-16
Objective To investigate the effect and molecular mechanism of Andrographolide (AD) on matrix metalloproteinase-9 (MMP-9) expression in human colon cancer H 3255 cells. Methods Human lung cancer H3255 cell line weve cultured in vitro, and treated with 1.0, 3.0, 5.0μmol/L AD for 24 h, untreated cells was used as blank control. Cell viability, cell migration and cell invasion were analyzed by MTT assay, scratch healing assay and transwell membrane assay, respectably. Expression of MMP-9 mRNA was analyzed by RT-PCR. Protein expression and phosphorylation of Akt were detected by Western blot. Activity of NF-κB and MMP-9 were analyzed by luciferase reporter assay. Results AD could significantly reduced H 3255 cells invasion and migration without affecting the viability of cells, as demonstrated by scratch healing and transwell membrane assay. Furthermore, Western blot and RT-PCR results showed that AD could markedly inhibited MMP-9 activity and its expression in both protein and mRNA levels. AD could attenuated Akt’s phosphorylation and the activity of NF-κB. Moreover, LY 294002, an inhibitor of PI3 K, could significantly inhibited NF-κB transcriptional activity and MMP-9 expression. In addition, different concentrations of AD could inhibit the promote activity of MMP-9. Conclusion AD was a potential anti-invasive agent by inhibiting MMP-9 involved in PI3 K/NF-κB pathways.
2.miR-200b suppresses proliferation and induces apoptosis in non-small cell lung cancer cells by targeting DNMT3A
Weimin LUO ; Xiangyu LUO ; Jialong GUO ; Chengyi LIN ; Jun ZHANG
Tianjin Medical Journal 2016;44(8):984-988
Objective To investigate whether miR-200b suppresses proliferation and induces apoptosis of non-small cell lung cancer cells by targeting DNMT3A. Methods A qRT-PCR was employed for detecting the expression of miR-200b in different non-small cell lung cancer cells and human bronchial epithelial cells. A549 cells were transfected with miR-200b mimics, scramble, DNMT3A-siRNA and control-siRNA, respectively. The scramble and control-siRNA were served the negative control of miR-200b mimics and DNMT3A-siRNA, respectively. Western blot assay was conducted to detect the expression of DNMT3A protein in A549 cells. MTT and Annexin V/propidium iodide staining were employed to detect the proliferation ability and apoptosis rate of A549 cells. The effects of miR-200b mimics and DNMT3A-siRNA on the proliferation and apoptosis rate of A549 cells were compared between groups. Results Results of qRT-PCR showed that the expression of miR-200b was significantly down-regulated in A549, H1299, L78 and H460 cells than that of 16HBE cells. Among them, the most obviously reduction was found in A549 cells (P<0.05). Western blot assay showed that the level of DNMT3A protein was inhibited by restored miR-200b or knock-down DNMT3A in A549 cells. After transfection of miR-200b mimics or knock-down DNMT3A for 48 h, 72 h and 96 h, MTT showed that the OD values, which reflected the optical density of cell proliferation were significantly lower than those in the control group (P<0.05). Annexin V/propidium iodide staining showed that apoptosis rates of A549 cells after transfection of miR-200b mimics or knock-down DNMT3A were (23.33%±0.90%and 20.41%±0.70%), compared with the control group (5.28%± 0.55%and 5.68%±0.47%, P<0.01). Conclusion miR-200b suppresses cell proliferation and induces apoptosis by targeting DNMT3A in non-small cell lung cancer.
3.C-phycocyanin protects doxorubicin-induced myocardial cells injury by Nrf 2/heme oxygenase-1 pathway
Xiangyu LUO ; Weimin LUO ; Xuesong ZENG ; Jun ZHANG
Chinese Journal of Biochemical Pharmaceutics 2014;(2):30-32
Objective To observe the role of heme oxygenase (HO)-1 on the protective effect of C-phycocyanin (CPC) on doxorubicin (DOX)-induced myocardial cells injury by Nvf2/HO-1 pathway. Methods 60 SD rats were randomly divided into control group, DOX group, CPC group and tin protoporphyrin IX (SnPP, an inhibitor of HO-1) group. The control group was injected with normal saline injection,while the DOX group was administrated with doxorubicin by intraperitoneal injection in a cumulative dose of 15 mg/kg for two weeks. For the CPC rats, 20, 40 and 60 mg/kg of CPC was administrated. The level of creatine kinase (CK) and lactate dehydrogenase (LDH) were detected, and the activity of HO-1 and caspase-3 were also examined. Expression of HO-1 and activation of Nrf 2 were detected by Western blot. Results Compared with control group, serum levels of CK, LDH and Caspase-3 activity in DOX group were significantly increased(P<0.05), but HO-1 in cardiac muscle was only increased slightly. upregulation. Treatment with CPC could significantly ameliorated the CK, LDH and Caspase-3 activity, and markedly induce HO-1 expression and its activity. The reduction of CK, LDH and Caspase-3 activity by CPC could be reversed by treatment of the HO-1 inhibitor, SnPP. Furthermore, CPC sould also induce Nrf 2 activation. Conclusion The protective effect of CPC on doxorubicin-induced myocardial cells in jury via Nrf 2 induced HO-1 HO-1 expression.
5.Protective effect of supplementary taurine on spleen hematopoietic damage in left-half-body ionizing irradiated mice
Long YANG ; Xiangyu XU ; Chengji LUO ; Tianmin CHENG
Journal of Third Military Medical University 2003;0(21):-
Objective To investigate the effect of half-body ionizing radiation on the concentration of MDA, SOD in serum and the spleen index, CFU-S and the protective function of supplementary taurine (Tau) in half-body ionizing irradiated mice. Methods Kunming mice were randomly divided into 6 groups: normal control, total-body irradiation (TBI), total-body irradiation+Tau, left-half-body irradiation, half-body irradiation+Tau, and total-body shielding irradiation. The relevant indexes such as the spleen index, spleen colony forming units (CFU-S) were observed. The ionizing radiation was performed under ~ 60 Co ? ray with absorbed dose 8.0 Gy, dose rate 68.46 cGy/min. Taurine at dose of 500 mg/kg was injected intraperitoneally twice a day before irradiation, once 30 min before irradiation, and once 6 h after irradiation. Results In TBI group, the spleen index and CFU-S were decreased remarkably, MDA increased and SOD reduced in serum. The hematopoietic function of spleen was not improved by supplementary taurine (P
6.The applied research of ultrasound-guided percutaneous thrombin injection for the treatment of Iarogenic femoral arterial complexity pseudoaneurysms
Zuoyi YAO ; Feifei LUO ; Xiangyu ZHOU ; Fang WANG
Chongqing Medicine 2017;46(4):450-452
Objective The aim to assess the methodology and feasibility of ultrasound guided percutaneous thrombin injection(UGTI) for the treatment of Iarogenic Femoral Arterial Complexity Pseudoaneurysms(IFACP).Methods Thirty two iarogenic femoral arterial complexity pseudoaneurysms patients following femoral arerial puncture for arterial angiography were treated with UGTI.Twenty-three IFACP with 2 lobes,8 IFACP with 3 lobes,1 IFACP with 4 lobes.Under local anesthesia the lobe was pene trated by artery needle successively and thrombin jection was performed slowely into distal lobe with US guide precise localization.Dynamical observation was performed for the status of thrombogenesis and cavity plugging.US follow-up examination were performed after 24 h and 7 d.Results Reperfusion occurred in IFACP with 3 lobes after 24 h and UGTI failure.IFACPs with 4 lobes failure.Nothromboembolic,infectious,allergic complication soccurred.Conclusion UGTI is the first mothed for the treatment of IFACP.Precise localization and percutaneous can enhance the ratio of treatment of IFACPs and avoid the severe complications.
7.Pathogenic Bacteria and Drug Sensitivity of 374 Cases of Children with Staphylococcal Scalded Skin Syn-drome
Zhu WEI ; Xiangyu LIU ; Yongqi LUO ; Xieling HE ; Jianping TANG
China Pharmacist 2016;19(6):1099-1101
Objective:To understand the pathogen bacteria detection and drug sensitivity results in the children with staphylococ -cal scalded skin syndrome to provide data for the clinical treatment .Methods: Totally 374 children with staphylococcal scalded skin syndrome treated from January 2010 to June 2015 were selected .The children's wound secretion and blood samples were collected , and bacterial culture and drug sensitivity test were carried out .Results:Totally 223 pathogenic bacteria were detected out in the wound se-cretion samples;17 cases of blood culture were positive with the positive rate of 4.55%;187 strains of staphylococcus aureus were de-tected out;64 strains of MRSA were found out with the MRSA detection rate of 34.22% (64/187).The sensitivities of MSSA and MRSA to common antibacterial drugs were different .The susceptibility rates of MSSA and MRSA to vancomycin , teicoplanin , teicopla-nin and linezolid were all 100.00%.The sensitivity rates of MRSA to penicillin , oxacillin, piperacillin, piperacillin, cefoperazone so-dium, cefazolin, cefuroxime, cefoxitin, azithromycin and clindamycin were all zero .Conclusion: The pathogenic examination of staphylococcal scalded skin syndrome is very important , and antibiotics should be used reasonably according to the results of drug sensi-tivity.
8.Correlation of monocyte counts prior to the collection of recombinant human granulocyte colony-stimulating factor primed bone marrow grafts with the quantities of CD34~+ cells in the grafts.
Yingjun CHANG ; Mingrui HUO ; Xiangyu ZHAO ; Xiaohua LUO ; Xiaojun HUANG
Chinese Journal of Practical Internal Medicine 2001;0(05):-
Objective To investigate the correlation of monocyte counts prior to the collection of recombinant human granulocyte colony-stimulating factor(rhG-CSF)primed bone marrow grafts(G-BM)with the quantities of CD34+ cells in the grafts.Methods From June 2004 to July 2007,fifty-three healthy donors were treated with rhG-CSF[5 ?g/(kg?d)]injected subcutaneously for five consecutive days.Bone marrow grafts and peripheral blood grafts were harvested on the 4th and 5th day,respectively.The quantities of CD34+ cells and blood routine prior to the collection of the G-BM were determined by flow cytometry and blood analyzer XL2100,respectively.Results The monocyte counts in peripheral blood(PB)of the 53 healthy donors were 896?424 per microliter.The counts of CD34+ cells per microliter and quantities of total CD34+ cells in the bone marrow grafts were 84?45 and(8.22?4.84)?107,respectively.Pearson correlation analysis indicated that the counts of monocyte in the PB correlated positively with the counts of CD34+ cells per microliter(r=0.573,P
9.Expression and significance inflammatory of cytokine mediated by Gli1 in acute pancreatitis
Kun LAI ; Xuemei HE ; Lan LUO ; Fang WANG ; Xiangyu ZHOU
Chinese Journal of Immunology 2017;33(7):1057-1061
Objective:To investigate the expression and the significance of inflammatory cytokine IL-6 through regulating Gli1 in acute pancreatitis.Methods: In this study,C57 mice were randomly divided into three groups:control group,model group,inhibitor group.caerulein intraperitoneal injection induce acute pancreatitis model.Use HE staining and amylase to testify the model successfully.Use RT-qPCR,Western blot to detect the expression of Gli1 in the pancreas,liver,lung,kidney and intestine and ELISA method to detect inflammatory cytokines IL-6.Results: Compared with control group,the expression of Gli1 is higher in model group,then the expression of IL-6 increases in inhibitor group which uses Gant61 to suppress Gli1 compared with model group.Conclusion: Gli1 may involved in the process of the distant tissue injury and repair in acute pancreatitis and through regulate its downstream cytokines like IL-6 to play a protective role in acute pancreatitis.
10.Protective effect and mechanism of Rosa laevigata Michx on adriamycin-induced cardiotoxicity
Weimin LUO ; Yuefeng LIU ; Xiangyu LUO ; Jun ZHANG ; Chengyi LIN ; Jialong GUO
Chinese Journal of Biochemical Pharmaceutics 2014;(1):29-31,34
Objective To study the protective effect mechanism of Rosa laevigata Michx (RLM) on cardiotoxicity induced by adriamycin in rats. Method 30 SD rats were randomly into control group, doxorubicin group and RLM groups. The control group was injected with normal saline injection, while the model group was injected with adriamycin intraperitoneally at the dosage of 15 mg/kg every other day. For the RLM groups,1~5 g/kg RLM were given after adriamycin injection. The survival rate, plasma BNP was observed. Apoptosis of cardiomyocyte was detected by instituted-labeled DNA (TUNEL). The activity of GSH-PX, CAT and total SOD in the myocardium tissue were also observed. The expression level of CuZn-SOD , bcl-2 and bax were detected by real-time PCR. Results The survival rate was significantly improved in SD rats treated with RLM compared with that in the adriamycin group (P<0.01). The BNP level was increased when treated by adriamycin (P<0.01), and decreased after RLM administration (P<0.01). RLM could also upregulate the expression of the CuZn-SOD mRNA level, and enhance the activity of GSH-PX, CAT and T-SOD compared with that in adriamycin group. Adriamycin could induce myocardial cells apoptosis, as demonstrated by TUNEL. RLM could inhibit adriamycin-induced apoptosis, bax mRNA expression, and increase bcl-2 expression and bcl-2/bax ratio. Conclusion RLM exhibit some antioxidant activity through many stages, and the anti-apoptosis activity may be related to affect the expression of bax and bcl-2 expression.