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Objective:To observe the expressions of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells, and to elucidate the their effects in the development of malignant glioma.Methods:The samples of normal human brain tissue and malignant glioma tissue were obtained and used as normal control group and malignant glioma group, respectively.The Cos7cells and the malignant glioma LN443and U343cells were cultured;the Cos7cells were used as normal cell control group, and the LN443and U343cells as malignant glioma cell group.Western blotting method was used to detect the expression levels of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells.Results:In brain tissue, the expression levels of SENP1, SENP2and SENP6proteins in malignant glioma group were higher than those in normal control group (P＜0.05) .Compared with normal cell control group, the expression levels of SENP1, SENP2and SENP6proteins in the LN443and U343cells in malignant glioma cell group were significantly increased (P＜0.05) .Conclusion:SENP1, SENP2and SENP6proteins highly express in the malignant glioma tissue and cells, and they may play an important role in promoting the occurrence of malignant glioma.
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Objective:To investigate the effect of Schisandra chinensis polysaccharide(SCP)on the growth of brain tumor stem cells(BTSCs),and to clarify the mechanism of inhibiting the growth of BTSCs of SCP. Methods:The primary human glioma cells were cultured,then the BTSCs were isolated by CD133 immunomagnetic sorting.The neural stem cell surface markers CD133 and Nestin were detected by immunofluorescence assay.The proliferation rate of BTSCs was examined by MTT assay.Annexin V-PI analysis was used to analyze the apoptotic rate of BTSCs.The expression levels of Bax,Bcl-2 and Caspase-3 proteins in BTSCs in various groups were detected by ELISA assay.Results:The results of immunofluorescence staining showed that the expressions of CD133 and Nestin were positive in BTSCs.Compared with control group,the proliferation rates of BTSCs in 200,400 and 800 mg·L-1SCP groups were decreased,especially in 400 and 800 mg·L-1SCP groups(P<0.05).The results of Annexin V-PI analysis showed that the apoptotic rate of BTSCs in 800 mg·L-1SCP group was increased compared with control group(P<0.05).The ELISA results showed that the expression levels of Bax in 200,400 and 800 mg·L-1SCP groups were significantly increased(P<0.05),and the values of Bax/Bcl-2 were significantly increased(P<0.05);compared with control group,the Bcl-2 expression level in the BTSCs in 800 mg·L-1SCP group was decreased(P<0.05).The expression level of Caspase-3 protein in 800 mg·L-1SCP group was also significantly increased compared with control group(P<0.01).Conclusion:SCP could inhibit the growth of BTSCs,and the induction of apoptosis may be one of mechanisms.
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Objective To further understand the clinical manifestations and improve clinical diagnosis of patients with leptomeningeal metastasizing high-grade glioma.Methods Sixteen patients with leptomeningeal metastasizing high-grade glioma (WHO classification:grade Ⅲ-Ⅳ) in Department of Radiotherapy,the First Hospital of Jilin University from July 2010 to September 2015 were respectively analyzed.The pathological types included anaplastic gliomas (1),anaplastic oligodenastrocytoma (1),glioblastoma (12),small-cell glioblastoma (1),gliosarcoma (1).We reviewed the relative clinical manifestations of the patients,and further compared them with 163 patients with systemic malignant solid tumors at corresponding period.Results The median time from initial diagnosis to the diagnosis of leptomeningeal metastasis was 13.0 months (range 2-19 months).Plain and enhanced magnetic resonance imaging was obtained in all patients.The main radiographic characteristics included ependymal enhancement (11),leptomeningeal enhancement (3),nodules of implantation metastasis in spinal canal (1),cranial nerve enhancement (2),and ventricular dilatation (1).Eight patients received cerebrospinal fluid examination.The diagnosis of leptomeningeal metastasis in 15/16 patients was determined by radiographic findings.Comparing with leptomeningeal metastasis from systemic malignant tumors at the corresponding period,the incidence of headache in patients with high-grade glioma was significantly lower (6/16 vs 81.6％ (133/163);x2 =16.3,P ＜ 0.01);and the incidence of cranial nerve paralysis was also significantly lower (4/16 vs 56.4％ (92/163);x2 =5.79,P =0.016 1).The incidence of nerve root symptoms was lower than that of systemic malignant tumors,though without statistically significant difference (2/16 vs 26.4％ (43/163);x2 =1.49,P=0.222).Nine patients respectively received chemotherapy,intrathecal chemotherapy or intrathecal chemotherapy combined with whole brain radiotherapy.The median survival tine was 4.5 months (range 0.7-13.3 months).Conclusions The imaging examination played an important role in the diagnosis of high-grade leptomeningeal metastasizing glioma.Comparing with the systemic malignant solid tumors,the leptomeningeal metastasizing high-grade glioma had its unique clinical characteristics.
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BACKGROUND: Vascular endothelial growth factor (VEGF) is a very effective way to make tissue engineer bone vascularization.However, because of expensive and short half-life, VEGF cannot maintain effective concentration in blood after injection. To resolve the problem effectively, gene transfection technique is used in this experiment to transfer human VEGF into seed cells-mesenchymal stem cells (MSCs) of tissue engineer bone and to make it secrete VEGF which could vascularize bone.OBJECTIVE: To explore the possibility of human vascular endothelial growth factor 165 (VEGF165) to transfect rabbit MSCs, and establish the experimental foundation of angiogenesis tissue engineering organization and the treatment of ischemic disorders.DESIGN: Observation control trail.SETTING: First Hospital of Jilin University and Institute of Frontier Medical Sciences of Jilin University.MATERIALS: The experiment was conducted in the Key Laboratory (BSL-2) of Frontier Medical Sciences of Jilin University between June 2003 and August 2004. Health New Zealand white rabbits, 4.0-5.0 months old, weighing 2.5-3.5 kg, half male and half female, were provided by Animal Center of Jilin University. The rabbits were handled under asepsis and anesthetized condition,corresponding to the animal ethical standard. Medicine and reagents: Ham F12 culture media (Gibco, U.S), MTT (Sigma, U.S)PLXSNKDRp-VEGF165 and pcDNA 3.0 vectors were prepared in the present laboratory. ELISA detection kit (Jingmei company,Shenzhen), DH5 α, restriction endonucleases Barn H I, Xhol Ⅰ, Hind Ⅲ, EcoR Ⅰ and standard DNA molecule (Promega,U.S) were also used in this study.METHODS: Rabbits' MSCs were separated and cultivated. The pcDNA 3.0-hVEGF165 expression vector was constructed and identified, pcDNA3.0-VEGF165 eukaryotic expression vector was constructed, the vector was used directly to transfect MSCs. The cultural supernatant then was collected and the soluble protein of human VEGF gene expression was analyzed with ELISA method.The proliferation capability of human umbilical vein endothelial cells (HUVEC) stimulated by the supernatant was measured with MTT methods, untreated MSCs and pcDNA3.0 transfected MSCs were used as control groups.MAIN OUTCOME MEASURES: ① Result of restriction enzyme digestion and DNA sequencing of the recombinant plasmid pcDNA3.0-VEGF165;② the secretion of human VEGF165 proteins of the transfected MSCs analyzed by ABC-ELISA; ③ MTT method was used to detect the effects of MSCs culture supematant transfected with VEGF165 on HUVEC cells proliferation ability.RESULTS: ①Result of restriction enzyme digestion and DNA sequencing of the recombinant plasmid: The constructed plasmid was digested with Hind Ⅲ and XHol Ⅰ, and then two pieces fragments were isolated with agarose gel electrophoresis, which was accordance with expected results. And sequencing results showed that PeDNA3.0-VEGF165 eukaryotic expression vector was successfully constructed. ② ABC-ELISA method: Compared with the control group, concentration of human VEGF protein in the supernatant of the cultured cells increased significantly after the MSCs were transfected with pcDNA3.0-VEGF165 for 24, 48, 72 hours (P＜0.05).③ MTT method was used to detect the effects of MSCs culture supernatant transfected with VEGF165 on HUVEC cells proliferation ability. The results showed MSCs supematant transfected with VEGF165 (2%, 4%,8%, 16%, and 32%) had statistical significance in promoting HUVEC cells proliferation rate compared with the normal control (P＜0.05).CONCLUSION: Human VEGF gene can be successfully transfected into MSCs and expressed effectively.
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Objective:To observe the living cells on the surface of intraocular lenses and to compare the cellular numbers and morphology of prestaining to that of poststaining.Methods:Twenty adult pigment rabbit eyes were given posterior chamber intraocular lens implantation.Lenses were extracted and put into 1640 cell cultural liquid immediately on the 1 st,3 rd,7 th,14 th and 28 th day respectively after operatrion.The samples were observed under inverted phase contrast microscope subsequently.Results:We found cells deposited on the surface of lenses from 1 st to 28 th day after operation.These cells contracted and lost three-demensional appearance while its number was less than that of prestaining obviously.Conclusion:A part of cells remove from the surface of lenses during staining;however,we could count the number but could not affirm cellular types which would make error in cellular classification and counting.The fact suggests that we should discover new accurate and reliable methods to avoid mistakes during experiment.