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BACKGROUND:WIF-1 is a tumor suppressor gene. Promoter hypermethylation causes WIF-1 down-regulationin most tumors. DNA methylation inhibitor can lead to gene demethylation and restore its expression. OBJECTIVE:To observe the differences of tumor pathology and, WIF-1 mRNAand proteinchanges using WIF-1 or 5-aza-2'-deoxycytidine demethylation in animalmodels of osteosarcoma. METHODS:Murine osteosarcoma models were established and divided into three groups. In the control group, no treatment was given. In the 5-aza-2'-deoxycytidine group, an appropriate amount of 5-aza-2'-deoxycytidine was injected ineach mouse daily. In the WIF-1 group, an appropriate amount of Wnt/β-catenin signal transduction pathway inhibitor WIF-1 was injected in each mouse daily. Seven days after medication, the weight of nude mouse was weighed every 7 days. Short tumor diameter (a) and the long diameter (b) were measured. Therelative tumor volume was calculated. The relative growth rate of tumor was calculated at 7, 14, 21, 28 and 56 days. Four nude mice from ach group were sacrificed by puling the neck at 7, 14, 21, 28 and 56 days after medication. Tumor tissues were stripped and the weight of them was weighed. Pathological analysis of the tumor was conducted. The expression of WIF-1protein and WIF-1 mRNA was detected in osteosarcoma at 56 days after medication in the three groups. RESULTS AND CONCLUSION:(1) Compared withthe medication and control groups, the weight of nude mice was increased at 7, 14, 21, 28 and 56 days in the treatment group. No significant difference was found between the medication and control groups. (2) The tumor size was significantly smaler in themedication group than in the control group. WIF-1 mRNA and WIF-1 protein expression was increased in the medication group compared with the control group to different degrees. (3) Results suggested that WIF-1 gene promoter methylation is one of the mechanisms of the development of osteosarcoma. Use of WIF-1 or 5-aza-2'-deoxycytidine demethylation can inhibit tumor growth in animal models of osteosarcoma.
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BACKGROUND:The choice of schwannoma resection is strongly associated with whether the tumor was completely resected, whether stretch during resection injures spinal nerves, and final y with the prognosis of treatment. OBJECTIVE:To evaluate the spinal stabilization after laminectomy combining unilateral or bilateral nail-rod system for schwannoma in the spinal canal or intervertebral foramen. METHODS:A total of 48 cases of schwannoma in the spinal canal or intervertebral foramen of neck, chest and waist underwent laminectomy combining unilateral or bilateral nail-rod system. 34 cases in spinal canal received bilateral nail-rod system, and 14 cases in the intervertebral foramen received unilateral nail-rod system. RESULTS AND CONCLUSION:At 3 days and 3, 6, 12 months after internal fixation, radiograph results demonstrated that location of implants was good. Bone graft fusion was found. No spinal instability and vertebral slippage occurred. Neural functional score Bodford (1997) and quality of life score were significantly increased after treatment (P<0.01). Muscle strength assessed by Lovett grade was significantly elevated after treatment (P<0.01). Pain evaluated by Virtual Rescan grade was significantly lessened after treatment (P<0.01). Schwannoma was completely resected in 48 patients. After treatment, six patients affected leakage of cerebrospinal fluid. One case experienced infection of cerebrospinal fluid. One case had to undergo secondary operation due to the infection. Three cases received nerve root resection due to tumor erosion. These experimental results confirmed that laminectomy combining unilateral or bilateral nail-rod system for schwannoma in the spinal canal or intervertebral foramen has the advantage of the tumor ful y exposed to the operator, which can help completely resect schwannoma and effectively avoid spinal nerve injury. Even more important thing is that the spinal stability is reconstructed by unilateral or bilateral nail-rod system, which prevents the occurrence of vertebral slippage and vertebral destabilization. Long-term effect stil needs further research.
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Objective To know the current status of the household lavatories in the countryside of Guangxi Province, China and to present the scientific data for the government to make decision in the lavatory improvement in the countryside. Methods A stratified cluster sampling was used in the investigation and 2 414 peasant households in 241 villages were selected in 2006. Results The popularization rate of sanitary lavatory was 43.79% and 50.95% of the peasant households used the non-sanitary lavatory, 5.26% had no household lavatory. The main types of the sanitary lavatories used in the investigated area were firedamp tank-like and three-case-cesspool, accounted for 50.14% and 33.96% respectively. The rate of dejecta treatment for the non-sanitary lavatories was only 13.74%. As the money spent on the sanitary lavatory in each household was no less than 810 Yuan (RMB), maybe the popularization rate of the sanitary household lavatory could reach 50%. Conclusion Although the popularization rate of sanitary household lavatory in the countryside of Guangxi province significantly increased in recent years, it still does not fulfill the requirement for controlling the intestinal infectious diseases and parasitism in this area.
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[Objective]To study the difference in expression of RFC, GST-?, DHFRmRNA between human osteosarcoma U2-OS cell line and the MTX-resisitant variants U2-OS/R1-R3, and to investigate the significance in MTX resistance for human osteosarcoma. [Methods]Three resistant MTX human osteosarcoma cell lines were established by pulse exposure parental cell line(U2-OS) in gradually increased dose of MTX . The expression of RFC,GST-?,DHFRmRNA were assayed by real-time fluorescence quantitative polymerase chain reaction(FQ-PCR).[Results]Three MTX-resistant variants(U2-OS/R1-R3) were successfully established , the results of the FQ-PCR revealed that the MTX resistance was associated with the decreased expression of the RFC mRNA and increased expression of DHFR mRNA and GST-? mRNA.[Conclusion]The author investigated the MTX resistant mechanism of human osteosarcoma cell line at a gene level. The decreased expression of RFC mRNA and the increased expression of DHFR mRNA and GST-? mRNA participate in the MTX resistance in human osteosarcoma cell lines U-2 OS. This provides the evidence for exploring the MTX resistance mechanism in clinical osteosarcoma patients ,and helps to screen the patients who are insensitive to MTX chemotherapy.
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[Objective]To observe whether or not caffeine can improve the cyto-toxicity effect of methotrexate(MTX) on osteosarcoma cell line.[Method]Osteosarcoma cell(OS-732)were incubated with no drug,caffeine,MTX or MTX adding caffeine separately,and were named as control group,caffeine group,MTX group and mixing group respectively.The ratios of cells on G2M phase of each group were measured with flow cytometry after 48 h incubation and the cell inhibition ratios were measured with the MTT colorimetric analysis after 72 h incubation.[Result]The ratios(%)of cells on G2M phase were control group(23.210?0.416),caffeine group(23.120?0.440),MTX group(28.770?0.531)and mixing group(0.105?0.002)(P
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[Objective]To investigate the expression of Survivin and its relationship with expression of PTEN and cyclinD1.[Method]Immonohistochemical method(SP method)was used to detect the expression of Survivin,PTEN and cyclinDl in 40 osteosarcoma tissues and 20 ostanchondroma tissues.[Result]The expression rate of Survivin was significantly higher in osteosarcoma(62.5%)than that in osteochondroma(10.0%)(P
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<p><b>OBJECTIVE</b>To provide a highly efficient adenoviral vector Ad-CMV-hTGFbeta1 for the study of gene therapy for reversion of the intervertebral disc degeneration.</p><p><b>METHODS</b>A newly developed recombinant adenoviral vector construction system was used in the study. The cDNA of hTGFbeta1 was first subcloned into a shuttle plasmid pShuttle-CMV. The resultant plasmid was linearized by digesting with restriction endonuclease PmeI, and subsequently transformed into E.coli. BJ5183 cells with an adenoviral backbone plasmid pAdEasy-1. Recombinants were selected by kanamycin resistance and confirmed by restriction endonuclease analysis. Finally, the recombinant plasmid linearized by PmeI was transfected into 293 cells. Recombinant adenoviruses were generated within 2 weeks.</p><p><b>RESULTS</b>The recombinant adenoviral plasmids were cut by BamHI and PacI respectively, and the diagnostic fragments appeared in 0.8% agarose electrophoresis. The infected 293 cells showed evident cytopathic effect (CPE). The productions of PCR confirmed the presence of recombinant adenovirus. The expression of hTGFbeta1 was verified by immunohistochemical staining.</p><p><b>CONCLUSIONS</b>The successful generation of the adenoviral vector Ad-CMV-hTGFbeta1 and the confirmation of the interest gene expression make it possible for the experimental study of the reversion of the intervertebral disc degeneration by gene therapy.</p>