50 Gy in the upper neck as compared to those treated with seperate facial and neck fields to 0.05)Conclusions We suggest that patients with neck negative NPC should be treated with conjoint facio cervical lateral fields and prophylactic whole neck irradiation with more than 50 Gy to the upper neck.

Objective To study the enhancing effect of ultrasound plus microbubble on small interference RNA(siRNA)transfection in vitro and in vivo.Methods Human vascular epithelial growth factor(VEGF)siRNA with 2'deoxy modification(VEGF 2'-eoxy siRNA,VdsR)was used.The human CNE cells(fromnasopharyngeal carcinoma)line was used for in vitro cell-based experiments and in vivo mouse xenograft model.Two different microbubble agents.BR14 and Levovist.were used together with the RNA transfection reagent RNA-mate.ELISA and RT-PCR assays were used to assess VEGF gene expression.Immunohistochemical staining (IHC)was performed to assess CD31 expression in xenograft tumors.Results VdsR transfection in CNE cells abolished VEGF expression as determined by ELISA experiments.In the first mouse xenograft experiment,ultrasound exposure dramatically enhanced VdsR-mediated tumor inhibition.In the second mouse xenograft experiment,when VdsR was mixed with the microbubble reagents and then injected into xenografts,ultrasoundexposure significantly reduced tumor growth in BR14-mixed VdsR group but not in the Levovist-mixed VdsR group compared to the control.RT-PCR experiments demonstrated that VEGF expression in ultrasound-exposed tumors was significantly lower than that in the control.Meanwhile,VEGF expression in the tumor tissue treated by BRl4-mixed VdsR declined as compared with the controls.Tumor vascular density as measured by CD31 immunostaining was significantly decreased in ultrasound-exposed tumors compared to the control.Conclusions Ultrasound exposure and/or microbubble can significantly enhance delivery and the efficiency of VdsR-mediated anti-tumor effects,and should be a location-specific enhancement approach for siRNA-based anti-cancer therapy.

Objective To explore microsurgical treatment of tuberculum sellae meningiomas.Methods A retrospective analysis was made on 35 cases of tuberculum sellae meningiomas operated from January 2005 to July 2013 in neurosurgery department of Sun Yat-sen Memorial Hospital,surgical approach,removal rate,surgical effect and complications were analysed.Results All patients were accepted microsurgical treatment,twenty cases were operated via subfrontal approach,four cases via anterior interhemispheric approach,ten cases via pterional approach,one case via combined subfrontal and pterional approach.According to Simpson grade,grade Ⅱ,rection was achieved in 26 cases,grade Ⅲ in 4 cases and grade Ⅳ in 5 cases.The total rection rate was 85.7％.There were 28 cases with merger ision loss and visual field defects preoperate,twenty cases were improved after operation,five cases with no change,three cases aggravated.The visual improved rate was achieved 71.4％,there was no surgical mortality case.Conclusion The surround tissue of tuberculum sellae meningiomas is very import ant,microsurgical rection is the main treatment.The choice of surgical approach should according to tumor size,growth pattern,degree of impaired vision and surgeon experience.Family with microanatomy and skillfull microsurgical techique can make sure operation succes.

This study was aimed to explore the impact of activated carbon adsorption modified by different concentrations of nitric acid and ammonia,in order to examine the impact on adsorption of mannotriose by modified activated carbon.The activated carbon was processed by different concentrations of nitric acid and ammonia.And then,the adsorption capacity of benzene,iodine,methylene blue and the purification effects on mannotriose were measured.The results showed that the active carbon modified by nitric acid and ammonia had some changes of adsorptions for methylene blue,iodine and benzene.The purification effect of mannotriose with nitric acid-modified activated carbon was declined.The purification effect of mannotriose with ammonia-modified activated carbon was increased.It was concluded that the pore structure of activated carbon had been changed by nitric acid and ammonia.The adsorption capacity of nitric acid modified active carbon to mannotriose was declined.However,the adsorption capacity of ammonia modified active carbon to mannotriose was increased.It showed that ammonia modified active carbon was suitable for the purification of mannotriose.And the adsorption capacity of iodine reflected the adsorption capacity of mannotriose by active carbon.

GITRL (Glucocorticoid-induced tumor necrosis factor receptor ligand) has been recently identified as a novel inhibitor of osteoclastogenesis and hence called Osteostat. In this study, we expressed recombinant extracellular domain of GITRL protein in Escherichia coli and analyzed its bioactivity. Using an Eco31I enzyme-based restriction and ligation method, we obtained an E. coli-preferred DNA sequence coding for the extracellular domain of human GITRL. The DNA was cloned into expression vector pQE-30Xa that encodes a fusion tag of 6xHis before the insert. The resultant recombinant expression vector pQE/GITRL was subsequently transformed into E. coli strain M15[pREP4]. After induction with Isopropyl beta-D-Thiogalactoside (IPTG), the cells produced the fusion protein mainly in the form of inclusion bodies as identified by SDS-PAGE. The recombinant protein was purified by affinity chromatography through Ni-NTA column and recognized by anti-His polyclonal antibody using Western blotting analysis. Moreover, we established a simple, efficient and sensitive reporter gene-based method to detect the activity of the recombinant protein. The results showed that the target protein was biologically active.
Base Sequence ; Escherichia coli ; genetics ; metabolism ; Extracellular Space ; metabolism ; Humans ; Molecular Sequence Data ; Recombinant Proteins ; analysis ; biosynthesis ; genetics ; Tumor Necrosis Factors ; biosynthesis ; genetics

Objective To investigate the correlation between homocysteine (Hcy) and cerebrovascular hemodynamic accumulative scores in primary hypertension patients. Methods A cross-sectional survey was conducted in 2 767 patients with essential hypertension who were simultaneously tested for serum Hcy and cerebral vascular function in the health management/physical examination center in Chongqing General Hospital from October 2015 to March 2018. The prevalence of hyperhomocysteinemia (HHcy) was also explored. Differences between cerebrovascular hemodynamic accumulative scores and its abnormal rate among different Hcy levels were evaluated using the analysis of variance and χ2tests, and logistic regression was used to analyze the correlation between Hcy and cerebrovascular hemodynamic accumulative scores. Results The median level of Hcy in primary hypertension was 11.8 (9.3-15.0) μmol/L. HHcy prevalence was 25.15% (27.01% in men and 19.80% in women), which was higher in men than women (χ2=14.576, P<0.001) and was increasing with age (P<0.001). The proportion of stroke, proportion of taking hypotensive medications, age, fasting plasma glucose, systolic pressure, pulse pressure, and Hcy were significantly higher in the abnormal score (<75 points) group (P<0.001) than in the normal score (≥75 points) group. The average cerebrovascular hemodynamic accumulative score was 86.99±16.10 points. The score in the highest quartile of Hcy (77.91±16.10) was significantly lower than that in other quartiles. The abnormal score rate (<75 points) was 15.25% and was increasing with the Hcy level (χ2=13.986, P<0.001). Logistic regression showed that Hcy in the second, third, and highest quartiles observed in abnormal scores was, respectively, 1.913-fold, 2.045-fold, and 7.497-fold higher than that in the lowest quartile after adjusting the confounding factors. Conclusion Hcy may be an independent risk factor for abnormal cerebrovascular hemodynamic accumulative scores in primary hypertension. Cerebrovascular dysfunction should be closely monitored when Hcy was higher than 15 μmol/L.

OBJECTIVES: To explore the mechanism of transforming growth factor-β1 (TGF-β1) induce renal fibrosis. METHODS: Renal fibroblast NRK-49F cells treated with and without TGF-β1 were subjected to RNA-seq analysis. DESeq2 was used for analysis. Differentially expressed genes were screened with the criteria of false discovery rate<0.05 and l o g 2 F C >1. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed for differentially expressed genes. Genes encoding transcription factors were further screened for differential expression genes. Then, the expression of these genes during renal fibrosis was verified using unilateral ureteral obstruction (UUO)-induced mouse renal fibrosis model and a public gene expression dataset (GSE104954). RESULTS: After TGF-β1 treatment for 6, 12 and 24 h, 552, 1209 and 1028 differentially expressed genes were identified, respectively. GO analysis indicated that these genes were significantly enriched in development, cell death, and cell migration. KEGG pathway analysis showed that in the early stage of TGF-β1 induction (TGF-β1 treatment for 6 h), the changes in Hippo, TGF-β and Wnt signaling pathways were observed, while in the late stage of TGF-β1 induction (TGF-β1 treatment for 24 h), the changes of extracellular matrix-receptor interaction, focal adhesion and adherens junction were mainly enriched. Among the 291 up-regulated differentially expressed genes treated with TGF-β1 for 6 h, 13 genes (Snai1, Irf8, Bhlhe40, Junb, Arid5a, Vdr, Lef1, Ahr, Foxo1, Myc, Tcf7, Foxc2, Glis1) encoded transcription factors. Validation in a cell model showed that TGF-β1 induced expression of 9 transcription factors (encoded by Snai1, Irf8, Bhlhe40, Junb, Arid5a, Vdr, Lef1, Myc, Tcf7), while the expression levels of the other 4 genes did not significantly change after TGF-β1 treatment. Validation results in UUO-induced mouse renal fibrosis model showed that Snai1, Irf8, Bhlhe40, Junb, Arid5a, Myc and Tcf7 were up-regulated after UUO, Vdr was down-regulated and there was no significant change in Lef1. Validation based on the GSE104954 dataset showed that IRF8 was significantly overexpressed in the renal tubulointerstitium of patients with diabetic nephropathy or IgA nephropathy, MYC was highly expressed in diabetic nephropathy, and the expressions of the other 7 genes were not significantly different compared with the control group. CONCLUSIONS TGF-β1 induces differentially expressed genes in renal fibroblasts, among which Irf8 and Myc were identified as potential targets of chronic kidney disease and renal fibrosis.
Mice ; Animals ; Humans ; Transforming Growth Factor beta1/metabolism* ; Diabetic Nephropathies/pathology* ; Transcriptome ; Signal Transduction ; Kidney ; Ureteral Obstruction/pathology* ; Fibrosis ; Interferon Regulatory Factors ; Transforming Growth Factor beta/metabolism* ; DNA-Binding Proteins/metabolism* ; Transcription Factors/metabolism*