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Objective To analyze survival benefits of radiotherapy in patients with nasopharyngeal carcinoma (NPC) with distant metastases and analyze relevant prognostic factors.Methods Medical records of 329 patients newly diagnosed with metastatic NPC screened from the Surveillance,Epidemiology and End Results (SEER) database (199 of 329 patients received radiotherapy) between 2010 and 2015 were retrospectively analyzed.Overall survival (OS) and disease-specific survival (DSS) were calculated by Kaplan-Meier curve.The effect of different clinicopathological factors on the clinical prognosis of metastatic NPC patients was evaluated by logrank test and Cox regression analysis.Results The median follow-up time was 12 months.The 3-and 5-year OS rates were 27.4％ and 19.7％.The median OS was 17.9 months.Univariate analysis demonstrated that patients aged＜ 50 years,male,undifferentiated type,stage T3 or T4,positive regional lymph node,brain and liver metastases and 1-2 metastatic sites obtained OS and DSS benefits at 3 years after radiotherapy.Univariate and multivariate Cox analyses after propensity score matching showed that radiotherapy was an independent prognostic factor for metastatic NPC (OS,P=0.004;DSS,P=0.014).Besides,patients aged 60-69 years (OS,P=0.033;DSS,P=0.045),keratinizing squamous cell carcinoma (OS,P＜ 0.05;DSS,P＜ 0.05),stage T4 (OS,P =0.002;DSS,P =0.024),1-2 metastatic sites (OS,P =0.039;DSS,P =0.058),3-4 metastatic sites (OS,P =0.003;DSS,P =0.005) and no chemotherapy (OS,P=0.000;DSS,P=0.000) had poor OS and DSS,whereas sex,race and degree of differentiation exerted no effect on OS and DSS.Conclusions Radiotherapy can significantly improve the OS and DSS of patients with metastatic NPC.Prospective and randomized controlled studies are required to further explore the role of radiotherapy in the management of metastatic NPC.
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BACKGROUND:Previous studies have suggested that achaete-scute homology 1 (ASCL1) plays a key role in the neuronal commitment. Therefore, somatic cels may directly differentiate into neurons by gene transfection ofASCL1, which wil provide new therapeutic strategies for optic nerve regeneration. OBJECTIVE:To construct a recombinant adenovirus vector expressing ratASCL1 gene for further research ofASCL1 gene function. METHODS:The ratASCL1 gene and advenovirus shuttle plasmid (pYr-adshuttle-4) which contained enhanced green fluorescent protein (EGFP) reporter gene were cleaved by restriction endonucleaseXhoI andEcoR I. The target gene fragments were connected together to generate a recombinant plasmid pYr-ads-4-rat-ASCL1 and then transfected into E.coliDH5α. The plasmid was confirmed to be constructed as expectation by enzyme digestion and sequence reaction. The plasmid pYr-ads-4-rat-ASCL1 and pAd/PL-DEST were reconstructed by homologous recombination processes to obtain rat ASCL1 recombinant adenovirus vector. The plasmid pYrAd-rASCL1 was linearized byPac I and subsequently transfected into HEK293 cels for packaging and amplification. RatASCL1 gene in the recombinant adenoviruses were identified by PCR. Virus titer was determined by tissue culture infectious dose 50. Infection efficiency was monitored by EGFP expression. RESULTS AND CONCLUSION:Restriction endonuclease digestion and DNA sequencing showed that the recombinant adenovirus vector pAd-rat-ASCL1 was constructed correctly. The positive amplification bands of 862 bp could be seen in PCR analysis. The virus titer reached 2×1010 pfu/mL. Infection efficiency of recombinant adenovirus in HEK293 cels was more than 80%. The results indicate that the recombinant the adenovirus vector containingASCL1 with high titer and infection efficiency has been successfuly constructed, which can be helpful for further research of the function and clinical application ofASCL1 gene for optic nerve regeneration.
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BACKGROUND:Engineered hepatic tissue is considered a promising strategy for healing acute liver failure. But, there are series of hindrances in the construction of engineered hepatic tissues, including acquisition of vital hepatocytes, choice of scaffolds and culture system, and nutrition supply. OBJECTIVE:To construct three kinds of engineered hepatic tissues in hope to screen the optimal one for transplantation in acute liver failure. METHODS:After purification, amplification, bone marrow mesenchymal stem cells were induced to differentiate into hepatocyte-like cells which were co-cultured with acellular amniotic membrane, DACRON PATCH cardiovascular surgical patch, biological surgical patch, respectively to construct three kinds of engineered hepatic tissues. After 3 days of culture, morphological and functional detections were performed. RESULTS AND CONCLUSION:Rat bone marrow mesenchymal stem cells with higher purity were successful y harvested by using density gradient centrifugation and adherent methods, and then the cells were differentiated into hepatocyte-like cells. In the three kinds of engineered hepatic tissues, hepatocyte-like cells were found to be combined with the biological surgical patch to the maximum extent, and their combination exhibited stronger ability of urea synthesis and albumin secretion, which provides experimental basis for treatment of acute liver failure.
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BACKGROUND:The degradation of extracellular matrix, which is mediated by matrix metal oproteinases, plays a crucial role in the corneal neovascularization. Tissue factor pathway inhibitor 2, a new type serine proteinase inhibitor, can effectively inhibit the activity of matrix metal oproteinases. OBJECTIVE:To elucidate the effect of tissue factor pathway inhibitor 2 on the expressions of matrix metal oproteinases in keratocytes in vitro. METHODS:Rabbit keratocytes were primarily cultured and subcultured in vitro. Plasmid vector pBos-Cite-neo/TFPI-2 was transfected into keratocytes with Lipofectamine 2000. The positive cells were selected using G418. RESULTS AND CONCLUSION:The results of reverse transcription-polymerase chain reaction, western blot analysis and gelatinase zymography analysis showed that, expression of mRNA and protein of tissue factor pathway inhibitor 2 was upregulated in the transfected keratocytes (P<0.05), while activity of matrix metal oproteinase 1 and 2 was significantly decreased (P<0.05). Experimental findings indicate that that tissue factor pathway inhibitor 2 strongly inhibits the activity of matrix metal oproteinase 1 and 2 in keratocytes.
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Colostrum provides essential nutrients and immunologically active factors that are beneficial to newborns. Our previous work demonstrated that milk contains large amounts of miRNA that is largely stored in milk-derived microvesicles (MVs). In the present study, we found that the MVs from colostrum contain significantly higher levels of several immune-related miRNAs. We hypothesized that the colostrum MVs may transfer the immune-related miRNAs into cells, which contribute to its immune modulatory feature. We isolated colostrum MVs by ultracentrifugation and demonstrated several immune modulation features associated with miRNAs. We also provide evidence that the physical structure of milk-derived MVs is essential for transfer miRNAs and following immune modulation effect. Moreover, we found that colostrum powder-derived MVs also contains higher levels of immune-related miRNAs that display similar immune modulation effects. Taken together, these results show that MV-containing immunerelated miRNAs may be a novel mechanism by which colostrum modulates body immune response.
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Animals , Cattle , Female , Mice , Pregnancy , Cell Movement , Cell Proliferation , Colostrum , Metabolism , Cytokines , Metabolism , Liposomes , Chemistry , Metabolism , Macrophages , Allergy and Immunology , Metabolism , MicroRNAs , Allergy and Immunology , Metabolism , Milk , Allergy and Immunology , Metabolism , Phagocytosis , UltracentrifugationABSTRACT
BACKGROUND: Some studies have demonstrated that the degradation of extracellular matrix (ECM), which matrix metalloproteinases (MMPs) participates in, plays a key step in the corneal neovascularization (CNV). Tissue factor pathway inhibitor 2 (TFPI-2), a new type serine proteinase inhibitor found recently, can effectively inhibit the activity of MMPs. Whether TFPI-2 gene transfection can influence CNV is unclear.OBJECTIVE: To investigate the effect of TFPI-2 gene transfection on CNV.DESIGN: Randomized controlled experiment.SETTING: Laboratory for Department of Surgery, Wuhan Union Hospital; Central Laboratory, the Affiliated Third Hospital of Sun Yat-sen University.MATERIALS: This study was carried out in the laboratory for Department of Surgery of Wuhan Union Hospital and State Central Laboratory of the Third Hospital Affiliated to Sun Yat-sen University between June 2004 and March 2006. Sixty healthy purebred adult New Zealand rabbits of either gender, weighing 2.5 to 3.0 kg, were involved. Preoperatively, no obvious anterior segment ocular lesion was found by slit-lamp examination. pBos-Cite-neo/TFPl-2 was kindly gifted by Dr. Zhong Ren (Department of Hematology, Union Hospital). Peroxydase blocking agent, nonimmune goat serum,mouse anti-human MMP-1, 2 and 3 monoclonal antibodies, biotin labeled goat-anti-mouse IgG second antibody (Santa cruz Company) were used in this study.METHODS: Experimental intervention: Experimental rabbit models of CNV were created in each group by silver nitrate cautery. Then, the rabbit models were randomized into 3 groups and 20 rabbits for each group. Different reagents were subconjunctivally injected via many points in each group: saline in the group Ⅰ, empty vector in the group Ⅱ, plasmid encoding TFPI-2 in the group Ⅲ. Experimental evaluation: CNV growth was observed under the slit-lamp biomicroscope.The expression of TFPI-2 in each rabbit model was detected by reverse transcription-polymerase chain reaction (RT-PCR) method 2 weeks after modeling; the expression of MMPs in corneal tissue was detected by immunohistochemical method at 3,5,7,9 and 14 days after modeling.TFPI-2 gene expression was significantly higher in the group Ⅲ than in the group Ⅱ and group Ⅰ (P ＜ 0.01); The MMP-1, 2, 3 expressions in the corneal tissue were significantly lower in the group Ⅲ than in the group Ⅱ and group Ⅰ,respectively, especially MMP-1, 3.
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Objective Observe the effect and the indications of intermittent short veno-venuous hemofiltration(ISVVH) in severe acute pancreatitis(SAP). Methods APACHE II scores≥14 and fluids imbalance were respectively used to define as the indication of starting hemofiltration and ending hemofiltration. In 39 patients with SAP,19 underwent ISVVH(IS group),and the other 20 patients were not accepted hemofiltration (N group). APACHE II scores, Balthazar CT grades and the plasma levels of procalitonin(PCT), TNF-?, IL-6, IL-8, IL-1ra, IL-2 and IL-10 were observed. Results At admission and 2d after admission, APACHE II scores in IS group and N group were (13.8?3.1)and (17.8?3.2) ( P
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Objective To study the effect of single short (SS) veno-venous hemofiltration and intermittent short (IS) veno-venous hemofiltration on imbalance of pro- and anti-inflammatory cytokines in severe acute pancreatitis (SAP) patients. Methods There were 17 and 21 SAP patients indicated for hemofiltration respectively enrolled for (SS) veno-venous hemofiltration (SS group) and IS veno-venous hemofiltration (IS group).Twenty SAP patients admitted and indicated for hemofiltration but not receiving the management during the same period served as control. Plasma levels of TNF-?,IL-6,IL-8,and IL-10 were measured. APACHE Ⅱ scores and fluid balance were used to evaluate patients′ condition. Results APACHE Ⅱ scores in both SS and IS groups declined significantly at day one after admission(P