1. Inhibitory effects of melittin on osteosarcoma xenografts in nude mice and its action on vasculogenic mimicry
Tumor 2008;28(9):771-775
Objective: To investigate the antitumor effects of melittin on UMR-106 cells with osteosarcoma xenografts in nude mice and its action on vasculogenic mimicry (VM) in vitro and in vivo. Methods: A three-dimensional cell culture system was formed on matrigel coats in vitro. Inhibitory action of melittin on the VM of UMR-106 cells was observed in vitro. Osteosarcoma was orthotopically transplanted in nude mice. The mice were randomly divided into three groups: normal saline (NS)-treated group, melittin group (320 μg/kg) and DDP-treated group(2 mg/kg). The drug was administered intratumorally for 10 d. The tumor volume and weight inhibition ratio were calculated, respectively. The VM density in tumor tissues was detected by CD34 and PAS double staining in vivo. The expressions of hypoxia-inducible factor-1α(HIF-1α) and matrix metalloproteinase-2 (MMP-2) proteins were determined by immunohistochemical staining and Western blot. Results: Melittin broke down the mesh and round architectonic of VM in osteosarcoma in vitro. The tumor weight inhibition ratio was 38.92% and the volume inhibition ratio was 43.04%. The VM density and the expressions of HIF-1α and MMP-2 proteins were decreased in the melittin group than that in the NS-treated group (P <0.01). Conclusion: Melittin markedly inhibites the growth of osteosarcoma UMR-106 xenografts in nude mice. The mechanism may be due to down-regulation of HIF-1α and MMP-2 expression and the inhibition of VM.
2.Activity of CDK1 in S phase cell checkpoint
Xia GAO ; Haocheng LONG ; Zhijian PAN ; Zhixiong LONG ; Jianping GONG
Cancer Research and Clinic 2012;24(3):165-168
Objective To investigate the phosphorylation and dephosphorylation of CDK1 based on the specific cell cycle apoptosis in Molt-4 cells and active variety of CDK1 in cell cycle specific apoptosis.Methods The exponential phase of growth Molt-4 cells (the human acute lymphoblastic leukemia cell line) were induced with dose response and time course of Camptothecin (CPT).The specific cell cycle apoptosis was detected with API method,then cell apoptosis was verified with post sorting confocal method.Meanwhile,the phosphorylation and dephosphorylation of CDK1 were detected by the protein electrophoretic analysis (Western blot).Results The specific cell cycle apoptosis occurred on exponential phase of growth Molt-4 cells after CPT treatment.When Molt-4 cells occured S-phase apoptosis, the apoptosis cell phosphorylation of CDK1-Thr161 band was more narrow than that of control cells, the apoptosis cell phosphorylation of CDK1-Thr15 band almost had the same band with control cells.Conclusion Cell apoptosis frequently developed in different cell cycle phase. API assay is quick and efficient method to analyze specific cell cycle apoptosis. When cell apoptosis take place in S-phase,the phosphorylation activity of CDK1 is inhibited.
3.Recent progress in study onneuroprotection of phosphodiesterase 5 inhibitors
Fan XU ; Ming LEI ; Long LONG ; Qihai GONG ; Jianmei GAO
Chinese Pharmacological Bulletin 2017;33(5):603-606
In recent years,the role of phosphodiesterase 5(PDE5)has been highlighted in the development and progression of neurological disease.PDE5 inhibitors show significant effect of neruoprotection,which may be related with some effects such as resistance to stroke,anti-oxidation,inhibition of neuroinflammation and amelioration of cognitive deficits.Based on the domestic and overseas researches about PDE5,this review systematically summarized the neuroprotection of PDE5 and their related mechanisms.
4.Dual effect of Molt-4 cell lines onset with Roscovitine
Haocheng LONG ; Xia GAO ; Zhijian PAN
Cancer Research and Clinic 2007;19(z1):1-3
Objective To investigate the effect of Mlot-4 cells onset with Roscovitine (ROSC)as some Cyclin-dependent kinases(CDK),inhibitor.Methods The logarithmic growth phase Molt-4 cells treated with ROSC at a final concentration ranging between 1~20 μmol/L and harvested in different time point,DNA assay of single cells by flow cytometry was used to detect the effect of cell cycle arrest and Annexin-V/FITC assay was used to detect the effect of cell apoptosis. Results It showed that ROSC exerted strong inhibitory effect on proliferation and cell cycle progression of Molt-4 Accumulation of G2/M arrested cells starting 6 h after onset of 10 μmol/L and 20 μmol/L ROSC;at the same time,the cell apoptosis of Molt-4 would be detected,According with the time and concentration changing,the cell apoptosis rate would rise.Conclusion It is concluded that Roscovitine(ROSC)as some Cyclin-dependent kinases(CDK),inhibitor,It has dual effects to Molt-4 cells,not only the effect of cell cycle arrest but also the effect of cell apoptosis.
5.Upper airway morphology of non snorers.
Chinese Journal of Stomatology 2007;42(4):216-218
Adolescent
;
Adult
;
Age Factors
;
Aged
;
Aged, 80 and over
;
Body Weight
;
Humans
;
Laryngeal Muscles
;
anatomy & histology
;
Magnetic Resonance Imaging
;
Middle Aged
;
Palate, Soft
;
anatomy & histology
;
Pharynx
;
anatomy & histology
;
Sleep Apnea, Obstructive
;
pathology
;
Tongue
;
anatomy & histology
;
Young Adult
6.Dentistry for obstructive sleep apnea hypopnea syndrome.
Chinese Journal of Stomatology 2007;42(4):193-194
7.Expressions of complement factors C1q and C3 in retina with negative lens-defocused myopia in guinea pigs
Tingting, GAO ; Xue, YANG ; Qin, LONG
Chinese Journal of Experimental Ophthalmology 2015;33(3):237-240
Background High myopia is seriously harmful to visual function.To explore the mechanism of high myopia is significant for the prevention and treatment.Complement proteins C1q and C3 have emerged as critical mediators of synaptic refinement and plasticity,however,their effects in retina on myopia remain elusive.Objective This study was to investigate the expressions of complement factors C1q and C3 in the retina with negative lensdefocused myopia in guinea pigs.Methods The use and care of animals complied with the application of Peking Union Medical College Hospital Iaboratory Animal Welfare Ethics Committee.Twelve 3-day-old pigmented guinea pigs were assigned randomly to the lens-defocused group and the normal control group.The left eyes were covered by -10 D PMMA lens for 4 weeks as the defocused eye group,and the right eyes were covered using 0 D PMMA lens in the same way as the fellow control eye group.The right eyes of the normal guinea pigs were used as the normal control eye group.The refractive diopter of guinea pigs was examined by retinoscopy.The animals were sacrificed at the fourth week and the expressions of complement C1q and C3 in the retinas of guinea pigs were detected by Western blot.Results The diptors were (-1.21±0.71)D,(+2.46±0.75)D and (+1.75±0.50)Din the defocused eye group,normal eye group and the fellow eye group at the fourth week,showing a significant difference among the groups (F=51.55,P=0.69),and diopters were insignificantly different between the normal eye group and fellow eye group (q =2.62,P=0.08).However,the diopters of the defocused eye group were significantly higher than those of the fellow eye group (q =10.92,P<0.01).The expressions of C1q and C3 proteins in the retinas were significantly different among the defocused eye group,fellow eye group and normal eye group (C1 q:F=8.810,P =0.003;C3:F =14.490,P<0.001),and expression levels of C1q and C3 proteins in the defocused eye group were significant higher than those of the fellow eye group (C1q:q=4.14,P=0.01 ;C3:q=4.71,P=0.005) ;while no significant differences were found between the fellow eye group and the normal eye group (C1q:q =1.61,P =0.27; C3:q =2.82,P =0.070).Conclusions The expression levels of C1q and C3 up-regulate in the retinas with lens-defocused myopic animal model.Excessive complement activation in retinas may be involved in the development of myopia.
8.Protective role and immunoregulatory effect of CXCR3 in hepatic ischemia-reperfusion injury
Jun GAO ; Gong CHEN ; Long LYU
Journal of Clinical Hepatology 2014;30(8):790-794
Objective To investigate the role and action mechanism of chemokine (C-X-C motif)receptor 3 (CXCR3)in hepatic ische-mia-reperfusion injury (IRI).Methods Forty-eight mice were divided into operation group and sham-operation group.The operation group was treated to establish a mouse model of IRI.Liver tissues were obtained at 3,6,12,and 24 h after IRI,with 6 mice at each time point.The expression of chemokine (C-X-C motif)ligand 9-1 1 (CXCL9-1 1 )and their receptor CXCR3 were measured by real-time PCR and Western blot.The effect of CXCR3 was blocked by its specific antagonist C6.Hepatic injury was estimated based on the activity of hepatic transaminase and morphological indices.The distribution of subsets of infiltrating T cells was analyzed by flow cytometry.All data were expressed as mean ±standard deviation.Comparison between groups was made by one-way analysis of variance.Results Compared with the sham-operation group,the operation group had significantly upregulated expression of CXCL9-1 1 and CXCR3 at all time points after IRI (P<0.05).Blocking CXCR3 significantly protected liver function and morphology (P<0.05).Antagonist C6 significantly re-duced Th1 cell infiltration (P<0.01),but significantly increased Treg infiltration (P<0.01).Conclusion CXCR3 is an ideal therapeu-tic target in IRI treatment due to its relationship with immunoregulation.
9.Correlation of GGT with AFP and diagnostic value of GGT for hepatocellular carcinoma
Chun GAO ; Long FANG ; Shukun YAO
Journal of Clinical Hepatology 2014;30(9):921-925
Objective To analyze the correlation of gamma-glutamyl transpeptidase (GGT)level with alpha-fetoprotein (AFP)level and to re-evaluate the diagnostic value of GGT for hepatocellular carcinoma (HCC).Methods Four hundred and seventy-two patients with HCC or liver cirrhosis,who were hospitalized in China-Japan Friendship Hospital from January 2003 to June 2009,were included in the study.The correlation between GGT and AFP was analyzed by Spearman nonparametric test.The cut-off values for the two parameters were determined based on their receiver operating characteristics (ROC)curves,areas under the ROC curve (AUCs),sensitivity,and specifici-ty,and the diagnostic values were presented using their sensitivity,specificity,and correct index.Statistical analysis was performed using SPSS 17.0.Normally distributed continuous data were analyzed by independent-samples t test,while non-normally distributed continuous data were analyzed by Mann -Whitney U test.Categorical data were analyzed by Pearson chi -square test,continuity-corrected chi -square test,or Fisher’s exact test.Results Among 472 patients,224 were diagnosed with HCC,and 248 with liver cirrhosis.Compared with cirrhotic patients,HCC patients had a significantly higher GGT level (113 (58-254)U/L vs 38 (22-72)U/L,Z=-11.037,P<0.001)and a significantly higher AFP level (429.5 (15.7-1210.0)ng/ml vs 5.7 (3.4-18.2)ng/ml,Z=-10.157,P<0.001).A significant correlation was found between GGT and AFP (r=0.449,P<0.001).The AUC was 0.784 for GGT and 0.788 for AFP.The cut-off value was 60 U/L for GGT and 20 ng/ml for AFP.The sensitivity was 74.1%for GGT,71.8%for AFP,and 90.7%for a combina-tion of the two parameters,the specificity was 70.2%,77.6%,and 58.7%,respectively,and the correct index was 0.443,0.494,and 0.494,respectively.Conclusion GGT may be regarded as one biomarker for HCC,and its level is significantly correlated with AFP level. The diagnostic value of AFP may not be improved when used in combination with GGT.
10.Effects of TSG on H2O2-induced Apoptosis and Expressions of XIAP and p53 in HUVECs
Ying YANG ; Xiqiang GAO ; Shiyin LONG
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2015;(6):629-633
Objective To study the effects of tetrahydroxy stilbene glucoside(TSG)on H2O2‐induced apoptosis of human umbilical vein endothelial cells (HUVECs)and on the expressions of X‐linked inhibitor of apoptosis protein (XIAP)and p53.Methods HUVECs were cultured in vitro and divided into 6 groups:control group ,300 μmol/L H2O2 group ,1 μmol/L TSG+ 300 μmol/L H2O2 group ,10 μmol/L TSG+300 μmol/L H2O2 group ,100 μmol/L TSG+ 300 μmol/L H2O2 group ,30μmol/L Embelin+ 10 μmol/L TSG+ 300 μmol/L H2O2 group.The morphology of apoptotic cells was observed by Hoechst 33258 staining.The mRNA and protein expressions of XIAP ,p53 ,Caspase‐3 were detected by RT‐PCR and Western blotting , respectively.Results The number of apoptotic cells and the expression level of p53 were significantly increased while the ex‐pression level of XIAP was dramatically decreased in H2O2 group as compared with control group.The expression level of p53 and the number of apoptotic cells were down‐regulated while the expression level of XIAP was up‐regulated after treatment with 10 or 100 μmol/L TSG when compared with H2O2group.Moreover ,compared with those in 10 μmol/L TSGgroup ,the number of apoptotic cells and the expression of Caspase‐3 were significantly enhanced after pretreatment with 30 μmol/L Embelin for 6 h.Conclusion TSG can inhibit H2O2‐induced apoptosis of HUVECs by down‐regulating the expression level of p53 and up‐reg‐ulating the expression level of XIAP.