1.Effects of tamoxifen on volume-activated Cl-currents of nasopharyngeal carcinoma cells at different stages of the cell cycle
Haibing LUO ; Liwei WANG ; Jianwen MAO ; Chenggang JIAO ; Aihui FAN ; Sihuai NIE ; Pan LI ; Lixin CHEN
Chinese Journal of Pathophysiology 1986;0(02):-
gluconate in both G1 phase and S phase cells.The permeability of G1 phase cells to I-was higher than that in S phase cells,but to gluconate was lower than that in S phase cells.CONCLUSIONS: The density of the volume-activated Cl-current,the anion permeability of the channel and the sensitivity of the current to tamoxifen were different between the CNE-2Z cells in G1 phase and those in S phase.The results suggest that the expression of tamoxifen-sensitive,volume-activated chloride channels is differentiated at different stages of the cell cycle.
2.Effect of resveratrol on miRNA-106b in Alzheimer′s disease animal model
Danli KONG ; Zuguo ZHAO ; Yuqing HE ; Yuanlin DING ; Yanhong LUO ; Biyu LIANG ; Jin WANG ; Haibing YU
Chinese Journal of Neurology 2016;49(11):846-851
Objective To investigate the effect of resveratrol on miRNA-106b in Alzheimer′s disease ( AD ) animal model.Methods Fifty Kunming male mice were divided into five groups by completely randomized block sampling.The five groups included three dosage resveratrol groups , an AD model group and a control group.The AD models were established in one month prior to treatments. Subsequently, from the 31st day various doses of resveratrol were provided intragastricly for 60 days.Then the memory function was observed by the step-down test.Meanwhile, the varying expressions of APP , P62, ApoA1, miRNA-106b, ABCA1 were tested in each group to determine whether there is the binding site for miRNA-106b in APP 3′UTR sequence.Results Compared with the control group by step-down test, the memory function of the AD model group mice decreased in different degree , which in the drug treatment group was higher than that in the model group (P<0.05).Compared with the AD group, the expression of APP (1.131 ±0.035) in the drug treatment group was higher than that in the model group (0.652 ± 0.026), while the P62 (0.412 ±0.022) and ApoA1 (0.534 ±0.032) were lower than the model group ( all P<0.05 ).High and medium dose groups of resveratrol treatment reduced varying degrees of APP (0.733 ±0.018,0.929 ±0.019,F=177.733) levels, and increased P62(0.954 ±0.035,0.633 ±0.015, F=434.5 ) and ApoA1 ( 1.042 ±0.051, 0.824 ±0.034, F=286.582 ) levels ( all P<0.05 ).The expression of miRNA-106b (0.464 ±0.313) and ABCA1(0.293 ±0.042) in the model group was lower than that in the control group (miRNA-106b 1.064 ±0.032, F=238.159; ABCA1 0.781 ±0.027,F=341.61;both P<0.05).The miRNA-106b (0.843 ±0.034, 0.601 ±0.012) and ABCA1 (0.882 ± 0.025, 0.624 ±0.036) levels in the high, medium dose resveratrol treatment groups increased to different extent ( both P<0.05 ).After the drug treatment , luciferase reporter vector experiments showed that the APP 3′UTR sequence contains the binding site of miRNA-106b.Conclusions APP is one of the target genes of miRNA-106b.Resveratrol is capable of improving AD by enhancing the expression of miRNA-106b and down-regulating the target genes including APP , P62 and ApoA1.This provides a new theoretical basis for the clinical treatment of AD.
3.Effects of Plasmid Fibroblast Growth Factor-2 Magnetic Chitosan Gelatin Microspheres on Proliferation and Differentiation of Mesenchymal Stem Cells.
Xingpo DING ; Ming LI ; Yujiang CAO ; Qiong YANG ; Tongchuan HE ; Cong LUO ; Haibing LI ; Yang BI
Journal of Biomedical Engineering 2015;32(5):1083-1089
The purpose of this study is to investigate the effect of superparamagnetic chitosan FGF-2 gelatin microspheres (SPCFGM) on the proliferation and differentiation of mouse mesenchymal stem cells. The superparamagnetic iron oxide chitosan nanoparticles (SPIOCNs) were synthesized by means of chemical co-precipitation, combined with FGF-2. Then The SPCFGM and superparamagnetic chitosan gelatin microspheres (SPCGM) were prepared by means of crosslinking-emulsion. The properties of SPCFGM and SPIONs were measured by laser diffraction particle size analyser and transmisson electron microscopy. The SPCFGM were measured for drug loading capacity, encapsulation efficiency and release pharmaceutical properties in vitro. The C3H10 cells were grouped according to the different ingredients being added to the culture medium: SPCFGM group, SPCGM group and DMEM as control group. Cell apoptosis was analyzed by DAPI staining. The protein expression level of FGF-2 was determined by Western blot. The proliferation activity and cell cycle phase of C3H10 were examined by CCK8 and flow cytometry. The results demonstrated that both of the SPIOCNs and SPCFGM were exhibited structure of spherical crystallization with a diameter of (25 ± 9) nm and (140 ± 12) μm, respectively. There were no apoptosis cells in the three group cells. Both the protein expression level of FGF-2 and cell proliferation activity increased significantly in the SPCFGM group cells (P < 0.05). The SPCFGM is successfully constructed and it can controlled-release FGF-2, remained the biological activity of FGF-2, which can promote proliferation activity of C3H10 cells, and are non-toxic to the cell.
Animals
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Cell Differentiation
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Cell Line
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Cell Proliferation
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Chitosan
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Fibroblast Growth Factor 2
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pharmacology
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Gelatin
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Magnetite Nanoparticles
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Mesenchymal Stromal Cells
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drug effects
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Mice
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Microspheres
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Plasmids
4.Role of Cl~- in regulatory volume decrease of nasopharyngeal carcinoma cells
Lixin CHEN ; Liwei WANG ; Linyan ZHU ; Sihuai NIE ; Jin ZHANG ; Ping ZHONG ; Haibing LUO ; Bo CAI ; Pan LI ; Jaco TIM
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To clarify the role of Cl - in regulatory volume decrease (RVD) of nasopharyngeal carcinoma cells (CNE-2Z).METHODS: Analysis of living cell images was used to detect the volume changes following exposure to hypotonic solution. Iron replacement and block of iron channels were also applied in the present study. RESULTS: Extracelluar hypotonic treatment made the cells swell and induced RVD. The RVD was correlated positively to the swelling in the range of 160-230 mOsmol/L. Substitution of gluconate for Cl - in perfusing solutions markedly increased RVD. Depletion of cellular Cl - abolished, and chloride channel blockers inhibited RVD. CONCLUSION: Cl - is the key iron to establish the RVD in CNE-2Z cells. Activation of Cl - channels and Cl - efflux are the major mechanisms of RVD.
5.miR-181c inhibits glycolysis by targeting hexokinase 2 in cancer-associated fibroblasts.
Haibing LAN ; Liang LUO ; Xiefei QI ; Yuanqi GONG ; Yu CHEN
Journal of Southern Medical University 2015;35(11):1619-1623
OBJECTIVETo investigate the role of miR-181c in glycolysis of cancer-associated fibroblasts (CAFs) and explore the mechanism.
METHODSHuman lung CAFs and normal fibroblasts (NFs), isolated from fresh human lung adenocarcinoma tissue specimens by primary culture of tissue explants, were transfected with a miR -181c mimics, a miR-181c inhibitor, a siRNA siRNA-HK2 or the vector HK2-vector via Lipofectamine(TM) 2000. Quantitative real-time PCR was used to analyze the changes in miR-125b expression in the transfected cells; hexokinase-2 (HK2) protein expression in the cells was detected using Western blotting, and the cellular glucose uptake was assessed with 2-NBDG. Lactate production in the cells was examined and expression of HK2 mRNA was detected with dual luciferase reporter gene assay.
RESULTSNo obvious difference was found in the cell morphology between CAFs and NFs. Compared with the NFs, the CAFs showed obviously increased glucose uptake, lactate production and HK2 protein expression with decreased expressions of the miR-181 family (P<0.05). Transfection with the miR-181 inhibito- rsignificantly increased glucose uptake, lactate production and HK2 protein expression in the NFs. In CAFs, transfection with the miR-181 mimics caused significantly lowered glucose uptake, lactate production and HK2 protein expression of. Knockdown of endogenous HK2 by siRNA abolished miR-181 mimics-mediated decrease of glucose uptake and lactate production in CAFs, while transfection with miR-181 mimics suppressed HK2 overexpression-induced enhancement of glucose uptake and lactate production in NFs.
CONCLUSIONTransfection with miR-181 mimics can suppress glycolysis in CAFs by inhibiting HK2 expression.
4-Chloro-7-nitrobenzofurazan ; analogs & derivatives ; Adenocarcinoma ; pathology ; Deoxyglucose ; analogs & derivatives ; Fibroblasts ; drug effects ; Glycolysis ; Hexokinase ; antagonists & inhibitors ; Humans ; Lung Neoplasms ; pathology ; MicroRNAs ; pharmacology ; RNA, Messenger ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Transfection ; Tumor Cells, Cultured
6.Effect and mechanism of Asperosaponin Ⅵ on polarization of M1/M2 macrophages
Jinfang LUO ; Ming LIU ; Hong YANG ; Haibing QIAN
Chinese Journal of Immunology 2023;39(12):2566-2570,2576
Objective:To study the regulation and mechanism of Asperosaponin Ⅵ on polarization of M1/M2 macrophages.Methods:MTT assay was used to detect the effects of Asperosaponin Ⅵ on RAW264.7 cell viability.The levels of TNF-α and IL-6 in supernatant of RAW264.7 cells induced by lipopolysaccharide(LPS)were determined by ELISA.The content of nitric oxide(NO)in supernatant of RAW264.7 cells induced by LPS was determined by Griess method.The gene expression levels of TNF-α,IL-6,argi-nase-1(Arg-1),heme oxygenase-1(HO-1)and suppressor of cytokine signaling protein-2(SOCS2)were detected by fluorescence quantitative PCR.Western blot was used to detect the expression levels of iNOS and p-p65 protein.Results:In LPS induced RAW264.7 cells,Asperosaponin Ⅵ inhibited protein or gene expression levels of TNF-α,IL-6,iNOS and p-p65,and increased HO-1 gene expression.Asperosaponin Ⅵ inhibited NO secretion in RAW264.7 cells induced by LPS.Asperosaponin Ⅵ increased the gene expression levels of M2 macrophage markers Arg1 and SOCS2 induced by IL-4.Conclusion:Asperosaponin Ⅵ inhibited RAW264.7 macrophage polarization to M1 type and promote it polarization to M2 type,which can play its anti-inflammatory and immunomodulato-ry role by regulating M1/M2 macrophage polarization.
7.Risk Assessment of the Onset of Sleep-related Painful Erection
Haibing HU ; Kunkun ZHAO ; Yongyi CHEN ; Daosheng LUO ; Wenjun BAI ; Ping LI ; Li ZHANG
Journal of Sun Yat-sen University(Medical Sciences) 2024;45(1):161-170
ObjectiveSleep-related painful erections (SRPE) is a rare sleep disorder characterized by repeated awakening due to painful interruptions of penile erections during nighttime sleep, and its etiology is currently unclear. The purpose of this study is to explore the impact of potential risk factors on the incidence of SRPE. MethodsInformation was collected through questionnaires administered to patients who presented at the urology department and suffered from SRPE or did not suffer from SRPE. A total of 290 participants completed the study, including 145 controls and 145 cases. Logistic regression analysis was used to assess the impact of age, occupation, sleep initiation time per night, frequency of sexual intercourse per week, psychological status, erectile dysfunction, chronic prostatitis, prostate enlargement, lumbar spine disease, central nervous system disease, hypertension, diabetes and family history on the onset of SRPE. ResultsSingle-factor logistic regression analysis found that a history of chronic prostatitis, intellectual labor occupation, central nervous system disease, late sleep onset, frequency of sexual activity, and anxiety status might be related to the onset of SRPE. After incorporating these factors into a multivariate regression analysis model, it was found that having sexual activity ≥2 times/week (OR 95%CI = 0.326(0.179,0.592) and late sleep onset (after 24:00) (OR 95%CI = 0.494(0.265,0.918)might be protective factors for SRPE, while a history of chronic prostatitis(OR 95%CI = 3.779(2.082,6.859) might be a risk factor for SRPE. However, there was no significant statistical difference in the impact of central nervous system diseases and occupation on multivariate analysis. ConclusionChronic prostatitis and anxiety status may be independent risk factors for SRPE; having sexual activity ≥2 times/week and delaying sleep time appropriately may be independent protective factors.