Objective： This study is aimed at developing a simple and easy way to generate dendritic cells （DCs） from human peripheral blood monocytes （PBMCs） in vitro. Methods： PBMCs were isolated directly from white blood...Objective： This study is aimed at developing a simple and easy way to generate dendritic cells （DCs） from human peripheral blood monocytes （PBMCs） in vitro. Methods： PBMCs were isolated directly from white blood cell rather than whole blood and purified by patching methods （collecting the attached cell and removing the suspension cell）. DCs were then generated by culturing PBMCs for six days with 30 ng/ml recombinant human granulocyte-macrophage stimulating factor （rhGM-CSF） and 20 ng/ml recombinant human interleukin-4 （rhIL-4） in vitro. On the sixth day, TNF-alpha （TNFα） 30 ng/ml was added into some DC cultures, which were then incubated for two additional days. The morphology was monitored by light microscopy and transmission electronic microscopy, and the phenotypes were determined by flow cytometry. Autologous mixed leukocyte reactions （MLR） were used to characterize DC function after TNFα or lipopolysaccharide （LPS） stimulations for 24 h. Results： After six days of culture, the monocytes developed significant dendritic morphology and a portion of cells expressed CDIa, CD80 and CD86, features of DCs. TNFα treatment induced DCs maturation and up-regulation of CD80, CD86 and CD83. Autologous MLR demonstrated that these DCs possess potent T-cell stimulatory capacity. Conclusion： This study developed a simple and easy way to generate DCs from PBMCs exposed to rhGM-CSF and rhIL-4. The DCs produced by this method acquired morphologic and antigenic characteristics of DCs.展开更多
We demonstrate the feasibility of performing a systematic screen for human gene functions in Drosophila by assaying for their ability to induce overexpression phenotypes. Over 1 500 transgenic fly lines corresponding ...We demonstrate the feasibility of performing a systematic screen for human gene functions in Drosophila by assaying for their ability to induce overexpression phenotypes. Over 1 500 transgenic fly lines corresponding to 236 human genes have been established. In all, 51 lines are capable of eliciting a phenotype suggesting that the human genes are functional. These heterologous genes are functionally relevant as we have found a similar mutant phenotype caused either by a dominant negative mutant form of the human ribosomal protein L8 gene or by RNAi downregulation of the Drosophila RPL8. Significantly, the Drosophila RPL8 mutant can be rescued by wild-type human RPL8. We also provide genetic evidence that Drosophila RPL8 is a new member of the insulin signaling pathway. In summary, the functions of many human genes appear to be highly conserved, and the ability to identify them in Drosophila represents a powerful genetic tool for large-scale analysis of human transcripts in vivo.展开更多
Yamanaka factors （Oct3/4, Sox2, Klf4, c-Myc） are highly expressed in embryonic stem （ES） cells, and their overexpression can induce pluripotency in both mouse and human somatic cells, indicating that these factors...Yamanaka factors （Oct3/4, Sox2, Klf4, c-Myc） are highly expressed in embryonic stem （ES） cells, and their overexpression can induce pluripotency in both mouse and human somatic cells, indicating that these factors regulate the developmental signaling network necessary for ES cell pluripotency. However, systemic analysis of the signaling pathways regulated by Yamanaka factors has not yet been fully described. In this study, we identified the target promoters of endogenous Yamanaka factors on a whole genome scale using ChIP （chromatin immunoprecipitation）- on-chip in El4.1 mouse ES cells, and we found that these four factors co-occupied 58 promoters. Interestingly, when Oct4 and Sox2 were analyzed as core factors, Klf4 functioned to enhance the core factors for development regulation, whereas c-Myc seemed to play a distinct role in regulating metabolism. The pathway analysis revealed that Yamanaka factors collectively regulate a developmental signaling network composed of 16 developmental signaling pathways, nine of which represent earlier unknown pathways in ES cells, including apoptosis and cellcycle pathways. We further analyzed data from a recent study examining Yamanaka factors in mouse ES ceils. Interestingly, this analysis also revealed 16 developmental signaling pathways, of which 14 pathways overlap with the ones revealed by this study, despite that the target genes and the signaling pathways regulated by each individual Yamanaka factor differ significantly between these two datasets. We suggest that Yamanaka factors critically regulate a developmental signaling network composed of approximately a dozen crucial developmental signaling pathways to maintain the pluripotency of ES cells and probably also to induce pluripotent stem cells.展开更多
Objective： The aim of our studywas to discuss changes in the clinicopathological characteristics of gastric cancer in Hehuang Valley of China in recent 10 years. Methods： A retrospective case-control study was perfo...Objective： The aim of our studywas to discuss changes in the clinicopathological characteristics of gastric cancer in Hehuang Valley of China in recent 10 years. Methods： A retrospective case-control study was performed on 2379 newly-diagnosed gastric cancer patients. All of them came from Hehuang Valley. The patients were divided into two groups [recent 5 years （R5Y） and late 5 years （L5Y）] from February 2003 to February 2013, and the clinicopathological data were surveyed retrospectively. Results： The constituent ratio of upper 1/3 gastric cancer in R5Y and L5Y was 33.5% and 20.7%, respectively, and it showed a significant difference between the two groups （X2 = 21.28, P = 0.00）, The constituent ratio of squamous carcinoma/adenosquamous carcinoma was 2.7% and 1.6%, respectively, and it also showed a significant differ- ence between two groups （X2 = 50.91, P = 0.00）. The constituent ratio of moderately-poorly differentiated/poorly differentiated gastric carcinoma was 84.2% and 50.2%, respectively, and it showed statistically significant difference （X2 = 30.28, P = 0.00）. The detection rate of early gastric cancer （EGC） was 1.47% （35/2379）. The constituent ratio of the types of Borrmann II and Borrmann IV of advanced gastric cancer （AGC） among 2379 cases was 47.6% and 40.8%, respectively, and that still showed statistically significant difference （X2 = 18,80, P = 0.00）. The constituent ratio of diffuse-type of gastric cancer in R5Y and L5Y was 36.2% and 30.8%, respectively, even there was a significant difference （X2 = 7.49, P = 0.01）. Furthermore, there were also significant differences in regional lymph nodes metastasis, perineural infiltration and vascular invasion （P 〈 0.05）. The positive detection rate of HER2, ER and PR was 14.88%, 17.23% and 15.93%, respectively. The constituent ratio of HP in two groups was 43.8% and 36.2% respectively, and it also showed a significant difference （~2 = 13.51, P = 0.00）. Conclusion： The pathogenic sites in gastric cancer change to the upper stomach in Hehuang Valley in recent 10 years, and the detection rate of squamous carcinoma/adenosquamous carcinoma reveals a sharp rise. Borrmann III is still one of the main types of advanced gastric cancer, but the detection rates of Borrmann II and IV are increasing. The main type of gastric cancer is the intestinal- type, but the ratio of diffuse-type is also increasing in recent 10 years. The HP detection rate is 40.65% （967/2379）, and it has a slight rise in recent 10 years. The detection rate of poorly differentiated adenocarcinoma is increasing despite the fact that the moderately/moderately-poorly differentiated adenocarcinoma is the main histologic types. High detection rates of lymph nodes metastasis, perineural infiltration and vascular invasion indicate poor prognosis in patients with gastric cancer. There is no change in HER2 positive rate, on the contrary, there are a little increase in ER and PR expression in Hehuang Valley.展开更多
The microstructural observation,the mass loss test,potentiodynamic polarization measurements and corrosion morphology examinations were conducted to study the influence of microstructural characteristics on corrosion ...The microstructural observation,the mass loss test,potentiodynamic polarization measurements and corrosion morphology examinations were conducted to study the influence of microstructural characteristics on corrosion behavior of Mg–5Sn–3In alloys in Hank’s solution after extrusion.The results show that the corrosion rate of the as-cast alloy is similar to that of as-extruded alloy;however,the local corrosion susceptibility is greatly weakened in the as-extruded alloy,especially in the extrusion direction.The relatively uniform corrosion morphology of the as-extruded alloy is attributed to refined Mg_(2)Sn particles,uniform distribution of Mg_(2)Sn particles and favorable crystal orientation.Meanwhile,the cytotoxicity tests confirm that the Mg–5Sn–3In alloy exhibits cytotoxicity of Grade 0−1 for NIH3T3 cells,suggesting an acceptable cytotoxicity of this alloy in the vitro assay.展开更多
We report a rare case of hypereosinophilic syndrome （HES） presenting with intractable gastric ulcers. A 71-year-old man was admitted with epigastric pain. Initial endoscopic findings revealed multiple, active gastri...We report a rare case of hypereosinophilic syndrome （HES） presenting with intractable gastric ulcers. A 71-year-old man was admitted with epigastric pain. Initial endoscopic findings revealed multiple, active gastric ulcers in the gastric antrum. He underwent Helicobacterpylori （Hpylon） eradication therapy followed by proton pump inhibitor （PPI） therapy. However, follow- up endoscopy at 4, 6, 10 and 14 mo revealed persistent multiple gastric ulcers without significant improvement. The proportion of his eosinophil count increased to 43% （total count： 7903/mm3）. Abdominal-pelvic and chest computed tomography scans showed multiple small nodules in the liver and both lungs. The endoscopic biopsy specimen taken from the gastric antrum revealed prominent eosinophilic infiltration, and the liver biopsy specimen also showed eosinophilic infiltration in the portal tract and sinusoid. A bone marrow biopsy disclosed eosinophilic hyperplasia as well as increased cellularity of 70%. The patient was finally diagnosed with HES involving the stomach, liver, lung, and bone marrow. When gastric ulcers do not improve despite H pylon eradication and prolonged PPI therapy, infiltrative gastric disorders such as HES should be considered.展开更多
Objective： To fabricate polymeric nanocomposites with excellent photoluminescence, magnetic properties, and stability in aqueous solutions, in order to improve specificity and sensitivity of cellular imaging under a ...Objective： To fabricate polymeric nanocomposites with excellent photoluminescence, magnetic properties, and stability in aqueous solutions, in order to improve specificity and sensitivity of cellular imaging under a magnetic field. Methods： Fluoridated LnS＋-doped HAP （Ln3＋-HAP） NPs and iron oxides （lOs） can be encapsulated with biocompatible polymers via a modified solvent exaction/evaporation technique to prepare polymeric nanocomposites with fluoridated Ln3＋-HAP/iron oxide. The nanocomposites were characterized for surface morphology, fluorescence spectra, magnetic properties and in vitro cytotoxicity. Magnetic targeted cellular imaging of such nanocomposites was also evaluated with confocal laser scanning microscope using A549 cells with or without magnetic field. Results： The fabricated nanocomposites showed good stability and excellent luminescent properties, as well as low in vitro cytotoxicity, indicating that the nanocomposites are suitable for biological applications. Nanocomposites under magnetic field achieved much higher cellular uptake via an energy-dependent pathway than those without magnetic field. Conclusion： 1tie nanocomposites fabricated in this study will be a promising tool for magnetic targeted cellular imaging with improved specificity and enhanced selection.展开更多
文摘Objective： This study is aimed at developing a simple and easy way to generate dendritic cells （DCs） from human peripheral blood monocytes （PBMCs） in vitro. Methods： PBMCs were isolated directly from white blood cell rather than whole blood and purified by patching methods （collecting the attached cell and removing the suspension cell）. DCs were then generated by culturing PBMCs for six days with 30 ng/ml recombinant human granulocyte-macrophage stimulating factor （rhGM-CSF） and 20 ng/ml recombinant human interleukin-4 （rhIL-4） in vitro. On the sixth day, TNF-alpha （TNFα） 30 ng/ml was added into some DC cultures, which were then incubated for two additional days. The morphology was monitored by light microscopy and transmission electronic microscopy, and the phenotypes were determined by flow cytometry. Autologous mixed leukocyte reactions （MLR） were used to characterize DC function after TNFα or lipopolysaccharide （LPS） stimulations for 24 h. Results： After six days of culture, the monocytes developed significant dendritic morphology and a portion of cells expressed CDIa, CD80 and CD86, features of DCs. TNFα treatment induced DCs maturation and up-regulation of CD80, CD86 and CD83. Autologous MLR demonstrated that these DCs possess potent T-cell stimulatory capacity. Conclusion： This study developed a simple and easy way to generate DCs from PBMCs exposed to rhGM-CSF and rhIL-4. The DCs produced by this method acquired morphologic and antigenic characteristics of DCs.
基金We are grateful to Xizhi Ma, Junnian Zhou, Tianhong Xu, Xu Liu, Xu Ding, Yang Liu, Ying Peng, Congwu Chi, Yiying Shang, Mingyao Ying, Sheng Ding, Lei Sun, Lei Tian, Huanhu Zhu, Hua Huang, Hongmei Li, and Xiaomo Wu for cDNA constructs and partial transgenic work, and Lihui Zhou （East China University of Science and Technology, China） for scanning electron microscopy. We thank Duc Nguyen （Yale University, USA） for critical reading and editing of this manuscript. This work is supported by grants from the National Natural Science Foundation of China （Grant Nos. 30030080, 39970408 and 30470840）, National Basic Research Program of China （973） （Grant No. 2006CB806700）.
文摘We demonstrate the feasibility of performing a systematic screen for human gene functions in Drosophila by assaying for their ability to induce overexpression phenotypes. Over 1 500 transgenic fly lines corresponding to 236 human genes have been established. In all, 51 lines are capable of eliciting a phenotype suggesting that the human genes are functional. These heterologous genes are functionally relevant as we have found a similar mutant phenotype caused either by a dominant negative mutant form of the human ribosomal protein L8 gene or by RNAi downregulation of the Drosophila RPL8. Significantly, the Drosophila RPL8 mutant can be rescued by wild-type human RPL8. We also provide genetic evidence that Drosophila RPL8 is a new member of the insulin signaling pathway. In summary, the functions of many human genes appear to be highly conserved, and the ability to identify them in Drosophila represents a powerful genetic tool for large-scale analysis of human transcripts in vivo.
基金We thank Drs J Zhao, DS Li, L Xiao （Chinese Academy of Sciences, China）, Drs B Leo and H Wang （Agilent Technologies, USA） for helpful discussions and technical assistance, and Drs HK Mei and Y Qiu （GlaxoSmithKline, UK） for the DAVID analysis. This research was supported by the Ministry of Science and Technology （2005CB522406, 2006CB943900, 2007CB947904, 2007CB947100, 2009CB941100, and 2007CB948000）, National Natural Science Foundation of China （30621091, 30625014, 30623003, and 90713047）, Shanghai Municipal Commission for Science and Technology （07PJ14099, 06ZR14098, and 06DZ22032）, and the Chinese Academy of Sciences （KSCX2-YW-R-56 and 2007KIP204）.
文摘Yamanaka factors （Oct3/4, Sox2, Klf4, c-Myc） are highly expressed in embryonic stem （ES） cells, and their overexpression can induce pluripotency in both mouse and human somatic cells, indicating that these factors regulate the developmental signaling network necessary for ES cell pluripotency. However, systemic analysis of the signaling pathways regulated by Yamanaka factors has not yet been fully described. In this study, we identified the target promoters of endogenous Yamanaka factors on a whole genome scale using ChIP （chromatin immunoprecipitation）- on-chip in El4.1 mouse ES cells, and we found that these four factors co-occupied 58 promoters. Interestingly, when Oct4 and Sox2 were analyzed as core factors, Klf4 functioned to enhance the core factors for development regulation, whereas c-Myc seemed to play a distinct role in regulating metabolism. The pathway analysis revealed that Yamanaka factors collectively regulate a developmental signaling network composed of 16 developmental signaling pathways, nine of which represent earlier unknown pathways in ES cells, including apoptosis and cellcycle pathways. We further analyzed data from a recent study examining Yamanaka factors in mouse ES ceils. Interestingly, this analysis also revealed 16 developmental signaling pathways, of which 14 pathways overlap with the ones revealed by this study, despite that the target genes and the signaling pathways regulated by each individual Yamanaka factor differ significantly between these two datasets. We suggest that Yamanaka factors critically regulate a developmental signaling network composed of approximately a dozen crucial developmental signaling pathways to maintain the pluripotency of ES cells and probably also to induce pluripotent stem cells.
基金Supported by a grant from the Basic Research Foundation of Qinghai Province(No.2011-Z-730)
文摘Objective： The aim of our studywas to discuss changes in the clinicopathological characteristics of gastric cancer in Hehuang Valley of China in recent 10 years. Methods： A retrospective case-control study was performed on 2379 newly-diagnosed gastric cancer patients. All of them came from Hehuang Valley. The patients were divided into two groups [recent 5 years （R5Y） and late 5 years （L5Y）] from February 2003 to February 2013, and the clinicopathological data were surveyed retrospectively. Results： The constituent ratio of upper 1/3 gastric cancer in R5Y and L5Y was 33.5% and 20.7%, respectively, and it showed a significant difference between the two groups （X2 = 21.28, P = 0.00）, The constituent ratio of squamous carcinoma/adenosquamous carcinoma was 2.7% and 1.6%, respectively, and it also showed a significant differ- ence between two groups （X2 = 50.91, P = 0.00）. The constituent ratio of moderately-poorly differentiated/poorly differentiated gastric carcinoma was 84.2% and 50.2%, respectively, and it showed statistically significant difference （X2 = 30.28, P = 0.00）. The detection rate of early gastric cancer （EGC） was 1.47% （35/2379）. The constituent ratio of the types of Borrmann II and Borrmann IV of advanced gastric cancer （AGC） among 2379 cases was 47.6% and 40.8%, respectively, and that still showed statistically significant difference （X2 = 18,80, P = 0.00）. The constituent ratio of diffuse-type of gastric cancer in R5Y and L5Y was 36.2% and 30.8%, respectively, even there was a significant difference （X2 = 7.49, P = 0.01）. Furthermore, there were also significant differences in regional lymph nodes metastasis, perineural infiltration and vascular invasion （P 〈 0.05）. The positive detection rate of HER2, ER and PR was 14.88%, 17.23% and 15.93%, respectively. The constituent ratio of HP in two groups was 43.8% and 36.2% respectively, and it also showed a significant difference （~2 = 13.51, P = 0.00）. Conclusion： The pathogenic sites in gastric cancer change to the upper stomach in Hehuang Valley in recent 10 years, and the detection rate of squamous carcinoma/adenosquamous carcinoma reveals a sharp rise. Borrmann III is still one of the main types of advanced gastric cancer, but the detection rates of Borrmann II and IV are increasing. The main type of gastric cancer is the intestinal- type, but the ratio of diffuse-type is also increasing in recent 10 years. The HP detection rate is 40.65% （967/2379）, and it has a slight rise in recent 10 years. The detection rate of poorly differentiated adenocarcinoma is increasing despite the fact that the moderately/moderately-poorly differentiated adenocarcinoma is the main histologic types. High detection rates of lymph nodes metastasis, perineural infiltration and vascular invasion indicate poor prognosis in patients with gastric cancer. There is no change in HER2 positive rate, on the contrary, there are a little increase in ER and PR expression in Hehuang Valley.
基金the National Key Research and Development Program of China(No.2017YFA0403803)the National Natural Science Foundation of China(Nos.52022017,51974058,51525401,51927801,81974325)+1 种基金the Science and Technology Commission of Shanghai Municipality,China(No.18ZR1428700)the Liaoning Revitalization Talents Program,China(No.XLYC1808005).
文摘The microstructural observation,the mass loss test,potentiodynamic polarization measurements and corrosion morphology examinations were conducted to study the influence of microstructural characteristics on corrosion behavior of Mg–5Sn–3In alloys in Hank’s solution after extrusion.The results show that the corrosion rate of the as-cast alloy is similar to that of as-extruded alloy;however,the local corrosion susceptibility is greatly weakened in the as-extruded alloy,especially in the extrusion direction.The relatively uniform corrosion morphology of the as-extruded alloy is attributed to refined Mg_(2)Sn particles,uniform distribution of Mg_(2)Sn particles and favorable crystal orientation.Meanwhile,the cytotoxicity tests confirm that the Mg–5Sn–3In alloy exhibits cytotoxicity of Grade 0−1 for NIH3T3 cells,suggesting an acceptable cytotoxicity of this alloy in the vitro assay.
基金Supported by Chung-Ang University College of Medicine
文摘We report a rare case of hypereosinophilic syndrome （HES） presenting with intractable gastric ulcers. A 71-year-old man was admitted with epigastric pain. Initial endoscopic findings revealed multiple, active gastric ulcers in the gastric antrum. He underwent Helicobacterpylori （Hpylon） eradication therapy followed by proton pump inhibitor （PPI） therapy. However, follow- up endoscopy at 4, 6, 10 and 14 mo revealed persistent multiple gastric ulcers without significant improvement. The proportion of his eosinophil count increased to 43% （total count： 7903/mm3）. Abdominal-pelvic and chest computed tomography scans showed multiple small nodules in the liver and both lungs. The endoscopic biopsy specimen taken from the gastric antrum revealed prominent eosinophilic infiltration, and the liver biopsy specimen also showed eosinophilic infiltration in the portal tract and sinusoid. A bone marrow biopsy disclosed eosinophilic hyperplasia as well as increased cellularity of 70%. The patient was finally diagnosed with HES involving the stomach, liver, lung, and bone marrow. When gastric ulcers do not improve despite H pylon eradication and prolonged PPI therapy, infiltrative gastric disorders such as HES should be considered.
基金supported by National Natural Science Foundation of China (Grant No. 21506161, 31270019)National Key Basic Research Program of China (973 Program) (Grant No. 2011CB933100, 2011CB932402)+1 种基金Guangdong Natural Science Funds for Distinguished Young Scholar (Grant No. 2014A030306036)open funds from the Key Laboratory of Biomedical Materials in Tianjin
文摘Objective： To fabricate polymeric nanocomposites with excellent photoluminescence, magnetic properties, and stability in aqueous solutions, in order to improve specificity and sensitivity of cellular imaging under a magnetic field. Methods： Fluoridated LnS＋-doped HAP （Ln3＋-HAP） NPs and iron oxides （lOs） can be encapsulated with biocompatible polymers via a modified solvent exaction/evaporation technique to prepare polymeric nanocomposites with fluoridated Ln3＋-HAP/iron oxide. The nanocomposites were characterized for surface morphology, fluorescence spectra, magnetic properties and in vitro cytotoxicity. Magnetic targeted cellular imaging of such nanocomposites was also evaluated with confocal laser scanning microscope using A549 cells with or without magnetic field. Results： The fabricated nanocomposites showed good stability and excellent luminescent properties, as well as low in vitro cytotoxicity, indicating that the nanocomposites are suitable for biological applications. Nanocomposites under magnetic field achieved much higher cellular uptake via an energy-dependent pathway than those without magnetic field. Conclusion： 1tie nanocomposites fabricated in this study will be a promising tool for magnetic targeted cellular imaging with improved specificity and enhanced selection.
