Objective To explore the pathogenesis-related imprinted genes in osteosarcoma and investigate the role of imprinted genes expression in osteosarcoma pathogenesis.Methods The model of malignant transformed immortalized human osteoblastic cells was established and the malignant transformed cells were collected on day 40,55,70,90.The untreated cells passaged from the normal cells on day 90 served as control.After the total RNA extraction,the finally synthesized biotinylated cDNAs were hybridized to CapitalBio Genechip CAP-F0017.2 of the differentially expressed genes.Two genes were chosen at random to further confirm the array results using the SYBR Green real-time PCR in samples of control,day 40 and day 90. Results By an entrance limit of ≥2.0 or ≤0.5,ten imprinted genes which biological functions are mainly related with cell growth/maintenance and signaling transduction were detectable for notablely differential expression,including down-regulated genes of CD81,GRB10,NDN,MEST and up-regulated genes of H19,MKRN3,SLC22A1L,TSSC3,CDKN1C in the malignant transformation of immortalized human osteoblastic cells.The array results were further confirmed by the real-time RT-PCR.Conclusion Ten imprinted genes were detectable for notablely differential expression.Our results will promote our understanding of the molecular mechanisms of imprinted genes in the course of osteosarcoma pathogenesis.

0.05). In hFOB, the expression of P53 was not detected, and that of P63 was weakly positive. The expression of P53 and P63 in HOS transformed by niekel sulfate were higher than that in normal HOS (P

Objective To establish a malignant transformed human fetal osteoblastic human cells from the immortalized cell line hFOB1.19 in order to explore the molecular mechanism in tumorigenesis of osteosarcoma. Methods hFOB1.19 cells were treated sequentially by an initiated factor, N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) and a promoter, 12-O-tetradecanoyl phorbol-13-acetate (TPA). The features of malignancy of transformed cells were identified by cell morphology, DNA content analysis, colony forming frequency on soft agar and tumorigenetic test in nude mice. Results Continuous passaging after the treatments resulted in the formation of a few paramorph foci and exhibited in an extensively random orientation. The poly-ploid of DNA was 81.08% in experiment group, much higher than that in control group (55.03%). Compared with that of negative control cell, colony formation efficiency of transformed cells in semisolid agar showed a a significant increase. The transformed cells formed tumors subcutaneously in the nude mice, which were verified to be poorly differentiated osteosarcoma histopathological examination. Conclusion Mocking the process of malignant transformation of human cells, we establish malignant transformed immortalized human fetal osteoblastic human cells (hFOB1.19) model.

Objective To obtain anoikis-resistant cells from human osteosarcoma cell line hFOB1.19 and investigate its biological characteristics.Methods The osteosarcoma cell line we used was malignant transformed immortalized human fetal osteoblastic human cells(hFOB1.19)which was transformed in our lab(called transformant in this paper).We have used anti-adhesion cell culture method to mimic detachment of tumor cells from extracellular matrix(ECM)to obtain anoikis-resistant variants.Transmission electron microscopy and fluorescence microscopy were used to observe the morphological changes,flow cytometry was used to observe the apoptotic rate.The growth characteristic was observed by MTT.The migration characteristics was examined by transwell cell culture system.Results Typical apoptotic cells were seen after de-adhesion culture.The apoptotic rate of the transformant at 24,48 and 72 h showed a gradually increasing tendency.The apoptotic rate of anoikis-resistant cells was lower than that of the transformant at the same time point.Compared with the transformant,the anoikis-resistant variants had stronger ability in proliferation and in migration with statistical significance(P

·AIM: To evaluate the efficacy and safety of intravitreal triamcinolone acetonide(TA) as treatment for macular edema associated with retinal vein occlusion(RVO).·METHODS: The study group consisting 30 patients (30 eyes) with RVO combined with macular edema received intravitreal 4mg TA. Changes in best-corrected visual acuity (BCVA), intraocular pressure(IOP), examination with slit-lamp microscope, fluorescein angiography and optical coherence tomography(OCT) were observed during the follow-up. Statistical analysis was conducted with SPSS 12.0 software.·RESULTS: The visual acuity(VA) of all patients was significantly improved and the central macular thickness (CMT) was significantly relieved. There was no correlation between course, age, CMT before injection and the type of RVO. There was positive correlation between visual acuity before injection and after injection.·CONCLUSION: Intravitreal injection of TA is an easy-operated and safe therapy. After injection, macular edema can be rapidly relieved. VA at baseline is the predictor for the prognosis of VA. Some patients experience recurrence of macular edema between 3 to 6 months after injection.

OBJECTIVE In order to investigate the possible anti-tumor molecular mechanisms of gecko polypeptide mixture (GPM). METHODS RNA-seq technology was used to identify the differen?tially expressed genes of human hepatocellular carcinoma (HCC) HepG2 cells treated with or without GPM. The HepG2 cells were treated with different concentration of GPM (0, 0.1, 0.2, 0.3, 0.4 mg·mL-1) for 6 h, 12 h and 24 h, respectively. MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells. Western blot analysis was applied to observe the expression of apoptosis-related proteins in HepG2 cells. RESULTS The results showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner. We applied many analysis methods, including differen?tially expressed genes analysis, Gene Ontology (GO) enrichment analysis, KEGG pathway enrichment analysis, protein- protein interaction network analysis to screen out possible molecular mechanisms. ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM. GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The mechanism is closely related to ERs, which might be beneficial for clinical therapy of HCC. CONCLUSION GPM can inhibit cells proliferation and induce apoptosis in HepG2 cells. The gene expression profile of GPM in HepG2 cells was obtained. The present study revealed the potential anti-tumor mechanism of GPM.

OBJECTIVE To explore the role of gecko crude peptides (GCPs) in the proliferation, apoptosis, migration and lymphangiogenesis of human hepatocellular carcinoma cells (HepG2) and human lymphaticendothelial cells (HLECs) in vitro. METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the anti- proliferative effect of GCPs and siRNA-VEGF-C on HepG2 cells, Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis. The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay. The protein and mRNA expressions of vascular endo?thelial growth factor-C (VEGF-C) and CXC chemokine receptor-4 (CXCR4) were detected by q-PCR, immunofluorescence, Western blot. The protein expressions of the extracellular signal regulated kinase (ERKI/2), c-Jun N-terminal kinase (JNK), p38-mitogen activated protein kinases (p38 MAPK), serine/threonine kinase (Akt) and phosphatidylinositol- 3- kinase (PI3K) were detected by western blot. The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay. The protein and mRNA expressions of vascular endothelial growth factor receptor-3 (VEGFR-3) and stromal cell-derived factor-1 (SDF-1) were detected by q-PCR, Western blot. RESULTS GCPs and siRNA-VEGF-C inhibited HepG2 proliferation, invasion and migration, and the most obvious inhibitory effect was both synergistic effects. Thus, GCPs suppressed HLECs proliferation, migration and tube-like structure formationin a dose- dependent manner, and had inhibitory effect of tumor- induced lymphangiogenesis in vitro. Additionally, we found that GCPs and siRNA- VEGF- C decreased the expressions of MMP-2, MMP-9, VEGF-C, CXCR4, phospho-ERK1/2, phospho-P38, phospho-JNK and PI3K in HepG2 cells. Moreover, GCPs had a dose-dependent depressive effecton the expressions of VEGFR- 3, SDF- 1 in HLECs. CONCLUSION The low expression of VEGF- C mediated by siRNA-VEGF-C and GCPs inhibit tumor proliferation, invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C, and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.

OBEJECTIVE Gecko has been clinically used in China for many years. It has been proved that the gecko polypeptide mixture(GPM)extracted from gecko could inhibit the growth of multiple types of tumor cells.In order to investigate the possible anti-tumor molecular mechanisms of GPM,we used RNA-seq technology to identify the differentially expressed genes of human hepatocellular carci-noma(HCC)HepG2 cells treated with or without GPM.METHODS The HepG2 cells were treated with different concentration of GPM(0,0.1,0.2,0.3,0.4 mg·mL-1)for 6 h,12 h and 24 h,respectively.MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells.Western blot analysis was applied to observe the expres-sion of apoptosis- related proteins and endoplasmic reticulum stress (ERs)-related proteins in HepG2 cells.Flow cytometry was also applied to detect reactive oxygen species(ROS)generation.In this report, we showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner.We applied many analysis methods,including differentially expressed genes analysis,Gene Ontology(GO)enrichment analysis,KEGG pathway enrichment analysis,protein-protein interaction network analysis to screen out possible molecular mechanisms.RESULTS ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM.GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The GPM could induce ROS generation and up-regulate ERs-related proteins. CONCLUSION The present study revealed the potential anti-tumor mechanism of GPM.

Objective:To investigate the expression of SHIP1 in the patients with acute myeloid leukemia and its effect on the apoptosis of human leukemia cells.Methods:The expression of SHIP1 in the bone marrow of patients with acute myeloid leukemia was detected by Westem blot.U937 cells was transfected with SHIP1 expression vector (pEGFP-SHIP1 group) and empty vector control (pEGFP group) respectively,U937 cells without transfection were used as the control group.Flow cytometry was used to detect the apoptosis of the cells,the expression of SHIP1,Bcl-2,Bax,Akt,p-Akt were detected by western blot.Results:The expression of SHIP1 in the bone marrow of patients with acute myeloid leukemia was significantly lower than that of the normal human bone marrow SHIP 1 (P＜0.01).The SHIP1 and Bax expressions as well as the apoptotic rate ofpEGFP-SHIP1 group were significantly higher than those of the control group(P＜0.01),while the Bcl-2 and p-Akt expressions were significantly lower than those in the control group(P＜0.01).Conclusions:SH-P1 expression was down regulated in the bone marrow of patients with acute myeloid leukemia.SHIP1 could promote the apoptosis of human leukemia cells via Akt signaling pathway.

AIM:To explore the treatment methods and curative effect of amblyopia in children, and to analyze the effect of compliance on the treatment process.METHODS:The children with amblyopia treatment since January 1st 2010 in General Hospital of Jinan Military Area, were selected and completed follow-up for a total of 129 eyes of 78 patients, including 53 eyes of male in 31 cases and 76 eyes of female in 47 cases, the initial age of 3 to 12 years, the treatment were performed with accurate optometry, reasonable glasses and appropriate to cover the dominant eye based comprehensive therapy and compliance training of the parents and children.The treatment time was 12-36mo, followed up for 3a.The final therapeutic effect was observed and the difference of the stage curative effect between the children with different grade of compliance (good, moderate and poor grade) was compared.RESULTS:In the treatment of 36mo, 106 eyes were cured, 21 eyes were essentially recovered and the total cure rate was 98.4%, which was improved in 2 eyes (1.6%) and was ineffective in 0 eyes (0%);the total effective rate were 100%.The total cure rate was 69% in the treatment of 6mo and there was a significant difference in the cure rate among children with different compliance (overall comparison P<0.001, each comparison P<0.017).In the treatment of 12mo, the total cure rate was 89.1% and the cure rate were significantly different between children with the moderate compliance and the children with good compliance (P<0.05).CONCLUSION:Comprehensive treatment of children with amblyopia can achieve an ideal clinical efficacy.Compliance is an important factor influencing the treatment effect and the treatment process.The parents and children`s compliance training can shorten the course of treatment and improve the curative effect.