1.Influence of FOXC1 downregulating on biological behavior of nasopharyngeal carcinoma (NPC) 5-8F cell lines
Bolin CHEN ; Shijiang YI ; Peng LIU ; Lei OUYANG ; Xun LEI
The Journal of Practical Medicine 2016;32(4):527-530
Objective To investigate the Influence of FOXC1 downregulating on biological behavior of nasopharyngeal carcinoma (NPC) 5-8F cell lines. Methods Western blotting was used to detect the protein levels of FOXC1 in human immortalized nasopharyngeal cell lines NP69, nasopharyngeal carcinoma cell lines 6-10B with lower metastasis potential and nasopharyngeal carcinoma cell lines 5-8F with higher metastasis potential. siRNA was used to downregulate the expression of FOXC1 in 5-8F cell lines, and then ability of migration and invasion were observed. Results FOXC1 protein expression level in NP69 cell lines, 6-10B cell lines and 5-8F cell lines were (0.27 ± 0.04), (0.7 ± 20.06), (1.08 ± 0.05) respectively, and the difference was statistically significant (P < 0.05). Also ability of cell migration and invasion significantly weakened after FOXC1 depletion in 5-8F cell lines (P < 0.05). Conclusion FOXC1 might regulate invasion and metastasis of nasopharyngeal carcinoma through changing expression of Fibronectin and Vimentin, and FOXC1 may be an available target for molecular target therapy of nasopharyngeal carcinoma.
2.The influences of long-term cryopreservation on cell viability and lymphocyte phenotypes of peripheral blood HS/PC collections.
Lei JIANG ; Jian OUYANG ; Junhao CHEN ; Al ET ;
Chinese Journal of Practical Internal Medicine 2006;0(S1):-
Objective To study the influences of long-term cryopreservation on nucleated cell viability,colony-form- ing units-granulocyte-macrophage(CFU-GM)-forming capability and Lymphocyte immune-phenotypes of pe- ripheral blood hematopoietic stem/progenitor cell(HS/PC)collections.Methods Fresh samples(n=12)were as control,frozen samples were divided into two groups:group 1(cryopreserved for 3-5 years,n=26)and group 2 (cryopreserved for 5-7 years,n=9).Flow cytometry(FCM)method based on 7 AAD or caspase-3staining was ap- plied to assess the degree of cell necrosis or apoptosis in cryopreserved and red-cell lytic collection samples.Lympho- cyte immune-phenotypes(CD3/CD4/CD8/CD19/CD16+56,CD4/CIM5RA/CIMSRO,CD8/CD28)were also de- tected by FCM.CFU-GM culture was done to study the hematopoietic activity.Results Percentages of necrosis and apoptosis cells of the control were both less than the groupl and group2(P
3.Separation,purification and preliminary structure analysis of Ramulus Mori polysaccharide
Peixiang WANG ; Zhen OUYANG ; Lei ZHANG ; Jing FANG ; Jianming TANG
Chinese Traditional Patent Medicine 1992;0(08):-
AIM: To study the isolation,purification and characterization of Ramulus Mori polysaccharide. METHODS: Ramulus Mori was extracted by boiling water. The raw extract was precipitated fractionally by alcohol deproteinized,passing through DEAE ion exchange cellulose ( DEAE-52) and SephadexG-100,obtained RMPS1 and RMPS2. The composition and characterization of Ramulus Mori polysaccharide were researched by TLC、IR、 GC、HPLC and smith degradation. RESULTS: The molecular weight of RMPS1 and RMPS2 were 5. 8 ? 105 and 6. 5 ? 105; RMPS1was made up of rhammose、arabinose、glucose and galactose with the molarity rate of 1. 08 ∶ 1 ∶ 1. 40 ∶ 1. 57; RMPS2 of rhammose、glucose and galactose with the molarity rate of 11. 38 ∶ 1 ∶ 1. 35. Smith degradation showed that the main linkage form in RMPS1 and RMPS2 was 1→2 and 1→4 glycosidic linkages,But some 1→3 glycosidic linkages also existed in the molecules; infrared spectrum showed that both had the polysaccharide characteristic absorption peaks. CONCLUSION: The structures of RMPS1 and RMPS2 are first determined from Ramulus Mori.
4.Effects of A Single Bout of Aerobic Exercise on Oxidative Stress and Total Peripheral Vascular Resistance
Feng LIANG ; Lei WANG ; Min CHAO ; Gang OUYANG ; Haiying GUO
Chinese Journal of Rehabilitation Theory and Practice 2015;21(5):586-590
Objective To explore the effects of a single bout of aerobic exercise on oxidative stress and total peripheral vascular resistance. Methods 36 prehypertensive patients were divided into aerobic exercise group (n=18) and control group (n=18). The aerobic exercise group performed cycle ergometry at 70% maximal oxygen consumption for 30 min. Total peripheral vascular resistance (TPVR), malondialdehyde (MDA), mean arterial pressure (MAP), vitamin C (VC) and vitamin E (VE) were measured before, immediately, 1 hour and 2 hours after the aerobic exercise. Results There was no significant difference in TPVR, MDA, MAP, VC and VE between two groups before exercise (P>0.05). TPVR was all significantly lower in the aerobic exercise group than in the control group immediately, 1 hour and 2 hours after exercise (P<0.001), and showed a rising trend of recovery. MDA was significantly higher in the aerobic exercise group than in the control group 1 hour and 2 hours after exercise (P<0.05), however there was no significant difference in MDA between two groups immediately after exercise (P>0.05). MAP was significantly lower immediately after exercise in the aerobic exercise group than in the control group (P< 0.001), but there was no significant 1 hour and 2 hours after exercise (P>0.05). There was no significant difference in VC and VE between two groups immediately, 1 hour and 2 hours after exercise (P>0.05). Conclusion The level of oxidative stress increases after a single bout of aerobic exercise, and the TPVR decreases.
5.Expression of FOXC1 and its relationship with E-cadherin in nasopharyngeal carcinoma tissues.
Peng LIU ; Shengkui TAN ; Shengjun XIAO ; Shijiang YI ; Lei OUYANG ; Qiaoyan HOU ; Weiming XIONG ; Xun LEI
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2014;28(15):1109-1112
OBJECTIVE:
To investigate the significance and relationship between the expression of FOXC1 and clinicopathological features, and to explore its correlation with E-cadherin.
METHOD:
Immunohistochemical SP method was used to detected the expression of FOXC1 in nasopharyngeal carcinoma tissues and nasopharyngitis tissues.
RESULT:
(1) Immunoreaction to FOXC1 was mainly located in nucleus of nasopharyngeal carcinoma cells. The positive expression rate of FOXC1 in nasopharyngeal carcinoma tissues was 85.3% (81/95), which was significantly higher than that in nasopharyngitis tissues (59.4%) (P < 0.05). (2) The expression of FOXC1 was not related to patients' age and gender, clinical stage of cancer and lymph node metastasis (P > 0.05). (3) There was a correlation between the expression of FOXC1 and down-regulated expression of E-cadherin in nasopharyngeal carcinoma tissues (P < 0.05).
CONCLUSION
FOXC1 may play an important role in generation and progression of nasopharyngeal carcinoma, there may be a correlation between the expression of FOXC1 and down-regulated expression of E-cadherin, also FOXC1 may play an important role in the process of EMT in nasopharyngeal carcinoma by regulating E-cadherin.
Adolescent
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Adult
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Aged
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Antigens, CD
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Cadherins
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metabolism
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Carcinoma
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Female
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Forkhead Transcription Factors
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metabolism
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Humans
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Male
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Middle Aged
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Nasopharyngeal Carcinoma
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Nasopharyngeal Neoplasms
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metabolism
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pathology
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Nasopharyngitis
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metabolism
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Young Adult
6.The study on the effect of predeposit autotransfusion in operation of lumbar disc protrusion
Lihua WANG ; Jinghui WU ; Dan ZHOU ; Hongwei SHAN ; Yanshuang SUN ; Lei TIAN ; Xilin OUYANG
Chinese Journal of Postgraduates of Medicine 2011;34(5):1-3
Objective To investigate the effect of predeposit autotransfusion in operation of the patients with lumbar disc protrusion.Methods Fifty patients of transfusion with lumbar disc protrusion were assigned into two groups by stratified sampling randomly,30 patients whose blood were predeposited before operation in experimental group,and the other 20 patients whose blood were not predeposited before operation in control group.The blood loss,the blood requirements during operations,the hemotological routine indexes and the complications related to blood transfusion were compared respectively.Results The blood loss of experimental group [ (720 ± 665 ) ml ] perioperative period was lower than that of control group [ ( 1060 ± 558 ) ml ],but there was no significant difference between two groups (P > 0.05 ).All the patients in experimental group went through perioperative period safely without allogenic blood transfusion.Hemoglobin,red blood cell and white blood cell were not significantly different between two groups before and after operation for 3,7 days (P> 0.05 ),the platelet count after operation for 7 days was significantly different between two groups (P < 0.05).No complication was observed in experimental group but 1 case with complication was observed in control group.Conclusions Predeposit autotransfusion is an effective to avoid homologous blood transfusion and its complications for the patients with lumbar disc protrusion.Furthermore,the clinical effect is not significantly different between the predeposit autotransfusion patients and the allogenic blood transfusion patients.
7.Effects of alarelin active immunity on FSHR expressions and bioinformatics of FSHR in ewes
Chiyue LEI ; Zhuandi GONG ; Min WEI ; Xiahui OUYANG ; Kun XIE ; Shien CHEN ; Suocheng WEI
Chinese Journal of Immunology 2014;(7):937-941
Objective:To investigate the effects of GnRH agonist on the expressions of FSHR proteins in pituitary and ovaries in ewes,also to analyse the bioinformatics characteristics of ovine pituitary FSHR.Methods: Forty-two prepubertal ewes (Ovis aries) were randomly assigned to six groups (n=7).The ewes in experimental group EG-Ⅰ,EG-Ⅱand EG-Ⅲwere subcutaneously injected with 200μg,300μg and 400 μg alarelin antigens twice (at day 0 and 14),respectively.Ewes in EG-Ⅳand EG-Ⅴwere injected sub-cutaneously with 200μg and 300μg alarelin antigen four times (at day 0,7,14 and 21),respectively.Ewes in the control group (CG) were subcutaneously injected with 2.0 ml solvent twice (at day 0 and 14).Fluorescence quantitative RT-PCR was used to detect gene expression of FSHR in pituitary glands.The expression of FSHR protein in the ovaries was detected using Western blot.Results:The 2-ΔΔCt values of GnRHR,FSHR and LHR mRNAs in the pituitary gland of EG were lower than that in control group (CG).The 2-ΔΔCt values in EG-Ⅳ( P<0.05 ) and EG-Ⅴ( P<0.05 ) were lower than those in EG-Ⅰand EG-Ⅱ.Expressions of FSHR proteins in EG ovaries increased.Levels of FSHR proteins in EG-Ⅳand EG-Ⅴ( P<0.05 ) were higher than CG.The length of sheep FSHR sequence of nucleotides were 1 091 bp,the homology of FSHR sequences was 100% with that reported in NCBI.The theory isoelectric point , half-life,unstable index,fat index,hydrophobic average were respectively 1.2 h,5.05,47.75,0.836 and 30.61.Conclusion:Alarelin active immunization can inhibit the expression of FSHR mRNA in the pituitary gland ,but increase FSHR expression in ovarian of ewes.FSHR is an unstable hydrophobic protein.
8.Correlations between resistin rs1477341, rs3219175 and type 2 diabetes mellitus in Guangdong
Meng LEI ; Xinqiang XU ; Shuting OUYANG ; Qiang ZHOU ; Bo CHEN ; Tianxing JI ; Zhen LIN
The Journal of Practical Medicine 2016;32(20):3296-3299
Objective To explore the relationships between resistin rs1477341 , rs3219175 and type2 diabetes mellitus (T2DM) in Guangdong. Methods 180 patients of diagnosed T2DM in the Second Affiliated Hospital of Guangzhou Medical University from January 2013 to December 2014 were enrolled in the study as the treatment group and another 180 subjects taking health examination as the control group. The genotype distributions of rs1477341 locus A/T, rs3219175 locus A/G were compared and the relationship between the genotypes and T2DM were anlysed by the logistic regression. Results The frequencies of the rs1477341, rs3219175 locus genotype reached the genetic equilibrium, showed their representativeness of a population group. The frequency distribution of rs1477341 genotype was statistically insignificant across the two groups (P > 0.05). The frequency of rs3219175 genotype AA+AG was decreased while genotype GG increased significantly compared to the control group (P < 0.05). The logistic regression results suggest genotype GG may increase disease risk of T2DM which the morbidity rate of AG homozygote was 0.630 times as high as that of GG homozygote (OR =0.630, 95%CI:0.405 ~ 0.980, P < 0.05) and the morbidity rate of GG homozygote was 1.551 times as higher as that of AA+AG homozygote (OR = 1.551, 95%CI:1.016 ~ 2.368, P < 0.05). Conclusions rs1477341 genotype has no correlation with T2DM among the population in Guangdong , but rs3219175 genotype does. GG may have a higher susceptibility to T2DM.
9.The effect of dexmedetomidine combined with ischemic preconditioning on liver ischemic reperfusion ;injury in rats
Youxiang GUO ; Kunhua QIU ; Lei ZHANG ; Yong LI ; Jiakun ZHANG ; Tingqiang WANG ; Jun OUYANG
Chinese Journal of Emergency Medicine 2016;25(9):1142-1148
Objective To investigate the effect of dexmedetomidine (Dex)combined with ischemic preconditioning on liver ischemic reperfusion injury in rats in order to explore its possible mechanism. Methods Sixty healthy male SD rats weighing (251 ±18)g,were randomly (random number)divided into five groups (n =12 in each):sham-operation group (Group S:operation without ischemia), ischemia-reperfusion group (Group IR:hepatic pedicle occlusion for 30 min and reperfusion for 6 h), dexmedetomidine preconditioning group (Group Dex: dexmedetomidine 25 μg/kg was given intra-peritoneally at 30 min before operation),ischemic preconditioning group (Group IP:10 min ischemia,10 min reperfusion,followed by hepatic IR)and Dex combined with ischemic preconditioning group (Group Dex +IP:Dex 25μg/kg was given intra-peritoneally at 30 min befor operation,10 min ischemia and 10 min reperfusion was given followed by hepatic IR).The hepatic inflow of blood stream was occluded for 30 min by Pringle maneuver to establish hepatic ischemic reperfusion injury (HIRI)rat model.At the end of reperfusion for 6 h,the concentration of ALT,AST and LDH (lactate dehydrogenase)in serum were measured.The liver histological changes were examined after HE staining.The liver cell apoptosis were examined by TUNEL. The expression of heme oxygenase-1 were examined by Westeren blot and immunohistochemistry.H2 O2 and GSH (r-glutamylcysteinylglycine ) in liver tissue were detected by spectrophotometer.Differences among the groups were analyzed by one-way analysis of variance (ANOVA) and Student-Newman-Keul test by using SPSS version 17.0 software.Differences were considered significant at P <0.05.Results Serum concentrations of ALT,AST and LDH in group IR,Dex,IP and Dex +IP were significantly higher than those in group S (P =0.000).And those biomarkers in group Dex,IP and Dex +IP,were significantly lower than those in group IR (P =0.000).Furthermore,those biomarkers in group Dex +IP,were significantly lower than those in group Dex and IP (P =0.000).There were no significant difference in the serum concentrations of ALT and AST between group Dex and IP (P =0.550, 0.771),and the serum level of LDH in group Dex was significantly lower than that in group IP (P =0.000).The liver histopathological score and apoptosis index were the lowest in group S and the highest in group IR,and those in group Dex and group IP were significantly lower than those in group IR (P =0.000),and those in group Dex +IP were significantly lower than those in group Dex and group IP (P =0.000),and there were no significant difference between group Dex and IP (P =0.704,0.661 ).The expression score of HO-1 was the lowest in group S and the highest in group Dex +IP,and that in group Dex and group IP was significantly lower than that in group Dex +IP (P =0.000,0.002),and that in group IR was significantly lower than that in group Dex and group IP (P =0.000),and there was no significant difference between group Dex and IP (P =0.099).In respect of H2 O2 level and GSH level in liver tissue, compared with group S,the H2 O2 levels in groups IR,Dex,IP and Dex +IP were significantly higher (P=0.000,0.000,0.000,0.001)while the GSH levels were significantly lower (P =0.000).Compared with group IR,the H2 O2 level in group Dex,IP and Dex +IP was significantly lower than that in group IR while the GSH level was significantly higher (P =0.000 ).The H2 O2 level in group Dex +IP was significantly lower than that in group Dex and IP while the GSH level was significantly higher (P =0.000).There were no significant difference between group Dex and IP (P =0.480,0.667).Conclusions Both of dexmedetomidine and ischemic preconditioning can protect liver from ischemia reperfusion injury in rats to some extent,and the combined application of two gives better effects,which is attributed to the increasing expression of HO-1 to a certain extent.
10.Experimental study of the anti-atherogenesis effects of sodium ferulate in hyperlipidemia rabbit and its mechanisms
Jingping OUYANG ; Baohua WANG ; Yongming LIU ; Jingwei YANG ; Lei WEI ; Ke LI ;
Chinese Pharmacological Bulletin 1987;0(02):-
AIM To study the anti atherogenesis action of sodium ferulate and its mechanisms. METHODS Atherosclerotic rabbit models were duplicated by feeding high lipid forage and ECV304 were cultured with the hyperlipidemic serum. The atherosclerotic plaque area was measured. Scanning electron microscope, spectrophotometer and immunocytochemical methods were used to detected the microstructures of endothelial cell, the content of NO in suspension and the expressions of TGF? 1, bFGF on the cell surfaces. RESULTS Sodium ferulate could decrease the plaque area, lessen the damnification of endothelial cell induced by HLS, enhance the expression of TGF? 1 and the release of NO from ECs, and reduce the expressions of bFGF in ECs, significantly. CONCLUSION Sodium ferulate can decrease the atherosclerotic plaque area induced by hypercholesterol, which may be relate to the expression change of cytokines.