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https://www.readbyqxmd.com/read/28109156/-cloning-and-sequence-analysis-of-variable-region-genes-of-mouse-anti-human-s100a9-monoclonal-antibody
#1
Min Yan, Danyang Mi, Xiangdong Sun, Zhenqiang Zhang, Huahui Zeng, Wendi Liu
Objective To clone and analyze variable region gene sequences of mouse monoclonal antibody against human S100A9 (anti-human S100A9 mAb). Methods Total RNA was extracted from the hybridoma cell line (IID4B10) secreting mouse anti-human S100A9 mAb. Furthermore, the heavy chain variable region (VH) and light chain variable region (VL) genes were amplified using reverse transcription-polymerase chain reaction (RT-PCR). After the cloning and sequencing of the VH and VL genes, the sequence analyses were performed by BLAST program...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109155/-preparation-and-application-of-rabbit-anti-mouse-setd8-polyclonal-antibody
#2
Haitao Ma, Jiaqian Guo, Jing Xia, Changmin Niu, Xueyi Shen, Hongya Sun, Ying Zheng
Objective To purify the recombinant Setd8 protein and prepare rabbit anti-mouse Setd8 polyclonal antibody. Methods The recombinant plasmid pET-30a-Setd8 was constructed by double enzyme digestion and linkage, and then transformed into E.coli BL21. The expression of the target protein was induced by IPTG and the expression product was purified by Ni-NTA affinity chromatograph. The purified protein was used to immunize New Zealand white rabbits to produce polyclonal antibody. The titer and specificity of the antibody were identified by ELISA, Western blotting and immunohistochemistry...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109154/-inhibitory-effect-of-overexpressed-kisspeptin-on-proliferation-and-migration-of-mkn-45-gastric-cancer-cells
#3
Xin Wang, Yijiong Li, Ruihua Zhang, Chao Pang, Xiaoqing Jiao, Zhenlong Zhu, Zhengmin Wang
Objective To investigate the expression of kisspeptin in gastric adenocarcinoma and its effect on the proliferation and migration of gastric cancer cells. Methods The level of kisspeptin in 50 gastric cancer tissues and their paracancerous tissues were detected by quantitative real-time PCR and Western blotting. Kisspeptin-siRNA, control-siRNA, pEGFP-N1-kisspeptin, pEGFP-N1 were separately transfected into MKN-45 gastric cancer cells. After 48-hour culture, the levels of matrix metalloproteinase-2 (MMP-2), β-catenin, C-myc, MMP-9, kisspeptin were detected by Western blotting; MTT assay was used to detect the proliferation of MKN-45 cells; wound healing assay was performed to assess the migration ability of MKN-45 cells...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109153/-chemotherapy-increases-fc%C3%AE-riib-content-in-peripheral-blood-of-the-patients-with-lung-squamous-cell-carcinoma
#4
Jingjing Lu, Xiuqin Cao, Zhiwei Yang
Objective To detect the FcγRIIB content of monocytes and B cells, the concentration of soluble FcγRIIB (sFcγRIIB) and its antibodies in the peripheral blood of patients with lung squamous cell carcinoma (LSCC) before and after chemotherapy. Methods The expression and localization of FcγRIIB on monocytes and B cells were detected by immunofluorescence technique; the expression of FcγRIIB mRNA was examined by quantitative real-time PCR; Western blotting was performed to detect the expression of FcγRIIB protein; the serum concentrations of sFcγRIIB, sFcγRIIB-IgG complex, anti-FcγRIIB autoantibody were determined by ELISA...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109152/-hepatic-stellate-cell-conditioned-medium-induces-proliferation-and-epithelial-mesenchymal-transition-via-activating-erk1-2-signaling-pathway-in-hepatoma-cells
#5
Yuxiao Xie, Rui Liao, Long Pan, Kai Fan, Cong Peng, Chengyou Du
Objective To investigate the influence of hepatic stellate cells (HSCs) on malignant biological behavior of hepatoma cells and related mechanisms. Methods Human hepatoma cell lines SMMC-7721 and HepG2, and hepatic stellate cell line LX-2 were cultured separately. HSC conditioned medium (LX2-CM), MAPK specific inhibitor U0126 were used to treat hepatoma cells, separately or together. The invasion and migration abilities of hepatoma cells were detected by Transwell(TM) assay, and cell proliferation was analyzed by CCK-8 assay...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109151/-tacrolimus-down-regulates-the-mrna-levels-of-il-17-and-il-23-in-the-muscle-tissues-of-mice-with-experimental-autoimmune-myositis
#6
Xiaojia Tian, Fangyuan He, Hongge Jia, Hongya Zhang
Objective To measure the expression levels of interleukin-17 (IL-17) and IL-23 mRNAs in the muscle tissue of the mice with experimental autoimmune myositis (EAM), and investigate the impact of tacrolimus (TAC) treatment on the mRNA levels of IL-17 and IL-23 and its therapeutic effect in EAM mice. Methods Fifteen female BALB/c mice were divided randomly into three groups, a normal control group, an EAM model group and a TAC-treated group. HE staining was used to observe the pathological changes for evaluating muscle inflammation of EAM mice...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109150/-isolation-culture-and-identification-of-adipose-derived-stem-cells-from-sd-rat-adipose-tissues-subjected-to-long-term-cryopreservation
#7
Qin Liu, Liping Wang, Fang Chen, Yi Zhang
Objective To study the feasibility of isolation and culture of adipose-derived stem cells (ADSCs) from SD rat adipose tissues subjected to long-term cryopreservation. Methods We took inguinal fat pads from healthy SD rats. Adipose tissues were stored with 100 mL/L dimethyl sulfoxide (DMSO) combined with 900 mL/L fetal bovine serum (FBS) in liquid nitrogen. Three months later, the adipose tissues were resuscitated for the isolation and culture of ADSCs. The growth status and morphology were observed. The growth curve and cell surface markers CD29, CD45, CD90 of the 3rd passage cells were analyzed respectively by CCK-8 assay and immunocytochemistry...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109149/-indirect-coculture-of-human-infrapatellar-fat-pad-derived-stem-cells-with-osteoarthritic-chondrocytes-induces-their-chondrogenesis
#8
Junjun Yang, Cheng Chen, Liu Yang, Xiongbo Song, Wenbin Xie, Shu Huang, Baorong Liu
Objective To promote phenotype recovery of osteoarthritic articular chondrocytes (OACs) and induce chondrogenic differentiation of infrapatellar fat pad-derived stem cells (IPFPSCs) by indirectly coculturing these two types of cells. Methods The OACs and IPFPSCs were isolated and cultured in vitro. This experiment included single IPFPSCs group, single OACs group, and coculture group. After cells were cultured in vitro with chondrogenic medium for 21 days, the chondrocyte phenotypes were determined by HE staining (cell morphology), Alcian blue staining (glycosaminoglycan content) and immunofluorescence cytochemistry (collagen 1, collagen 2, collagen 3, aggrecan, SOX9)...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109148/-rat-bone-marrow-mesenchymal-stem-cells-promote-apoptosis-and-inhibit-proliferation-and-migration-of-rsc96-cells
#9
Yongbin Ma, Suohua Ge, Dan Zhou, Huanyan Zhang, Luyang Sun, Xuefeng Wang, Jianhua Su
Objective To investigate the effect of rat bone marrow mesenchymal stem cells (BMSCs) on the proliferation and migration of rat RSC96 Schwann cells in vitro. Methods BMSCs of Sprague Dawley (SD) rats were isolated by bone marrow cell adherent method. The morphology of the third generation of BMSCs was observed by inverted microscopy. The cell surface antigen makers CD19, CD34, CD73, CD105 were detected by flow cytometry. The third-passage BMSCs differentiated into osteoblasts and adipocytes after exposed to bone-inducing and lipid-inducing media for 3 and 2 weeks, respectively, and then were identified by oil red O and alkaline phosphatase staining...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109147/-co-culture-with-umbilical-cord-mesenchymal-stem-cells-promotes-the-synthesis-and-mechnism-of-milk-protein-in-bovine-mammary-epithelial-cells
#10
Yankun Zhao, Wei Shao, Chenglong Luo, Kaile Wu, Xiong Yu
Objective To explore the possible mechanism of umbilical cord mesenchymal stem cells (UCMSCs) regulating milk protein synthesis in bovine mammary gland epithelial cells (BMECs). Methods UCMSCs and BMECs were co-cultured by double-chamber Transwell(TM), and other BMECs were cultured alone as a control group. Insulin like growth factor 1 receptor (IGF-1R) inhibitor AG1024 was used to treat cells. IGF-1, β casein (CSN2) and κ casein (CSN3) content in the supernatants were determined by ELISA; the relative expression abundance of Janus kinase and signal transducer and activator of transcription factor (JAK/STAT) signaling pathway-related genes were detected by quantitative real-time PCR (qRT-PCR); and after the cells were treated with JAK2 signal blocker AG490, qRT-PCR was performed to test the relative expression abundance of CSN2 and CSN3 mRNAs...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109146/-cloning-expression-and-activity-identification-of-human-innate-immune-protein-apolipoprotein-b-mrna-editing-enzyme-catalytic-subunit-3a-apobec3a
#11
Shan Cheng, Liyan Cao, Juan Du, Wenyong Wang, Yanhai Guo
Objective To construct the expression vector of apolipoprotein B mRNA editing enzyme catalytic subunit 3A (APOBEC3A), express APOBEC3A in eukaryotic cells and identify its cytosine deaminase activity. Methods The APOBEC3A gene was obtained by PCR and inserted into the eukaryotic expression vector pcDNA3.0(+). The recombinant vector pcDNA3.0-APOBEC3A was then transfected into HEK293T and HepG2 cells after confirmed by DNA sequencing. The recombinant protein was purified by Ni-NTA HisBind affinity column. Western blot analysis was used to detect the expression of APOBEC3A protein...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109145/-short-term-use-of-rapamycin-combined-with-tregs-prolongs-mouse-cardiac-graft-survival-but-attenuates-recipient-s-anti-tumor-immunity
#12
Ting Yan, Chen Luo, Xianmo Yang, Lei Ji, Shiqiao Luo
Objective To study the effect of short-term use of rapamycin(Rap) combined with regulatory T cells (Tregs) on the long-time survival of allogeneic mouse cardiac transplant, and its impact on the anti-tumor immunity of recipient. MethodsMouse Tregs were purified from recipients' spleen by magnetic activated cell sorting (MACS), and expanded by CD3/CD28 monoclonal antibody immunomagnetic beads and 2000 U/mL recombinant mouse IL-2 (rmIL-2) ex vivo. The purity was tested by fluorescence-activated cell sorting (FACS)...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109144/-interferon-regulatory-factor-5-irf5-regulates-the-differentiation-of-bone-marrow-derived-macrophages-in-mice
#13
Kang Sun, Jianguo Qu, Jixiang Chen, Shengchun Dang, Songbing He, Jin Zhang, Rong Xie, Yun Wang, Jianxin Zhang
Objective To investigate the expression differences of interferon regulatory factor 5 (IRF5) between M1 and M2 macrophages derived from mouse bone marrow and the effects of small interfering RNA targeting IRF5 gene (IRF5 siRNA) on macrophage differentiation. Methods With the treatment of IFN-γ and lipopolysaccharide (LPS), mouse bone marrow-derived macrophages were differentiated into M1 macrophages; while with the treatment of IL-4, mouse bone marrow-derived macrophages were differentiated into M2 macrophages...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109143/-decreased-number-and-immune-activity-of-splenic-t-lymphocytes-in-mice-exposed-to-hypoxia-at-high-altitude
#14
Xiaona Zhang, Jidong Li, Fangjie Hu, Shilan Gao, Xiaoyan Pu, Sheng Yong
Objective To explore the changes of splenic T lymphocyte subsets and functions in mice under high-altitude hypoxic conditions. Methods After mice were exposed to an altitude of 400 m, 2200 m and 4200 m for 30 days, ELISA was used to detect the concentrations of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in the cultured splenocyte supernatant; MTT assay was used to analyze the proliferation of splenic T lymphocytes; flow cytometry was performed to examine the alterations of splenic T lymphocyte subsets...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109142/-changes-and-role-evaluation-of-tnf-%C3%AE-and-il-1%C3%AE-in-lung-tissues-of-ards-mice
#15
Jianing Liang, Qianqian Zhou, Tianxiang Zhang, Xiaosu Wang, Liqiang Song
Objective To study the expression levels of TNF-α and IL-1β in the lung tissues of acute respiratory distress syndrome (ARDS) mice and their relationships with the severity of lung injury in the mice. Methods A mouse model of ARDS was induced by lipopolysaccharide (LPS). The morphological changes of lung tissue was observed by HE staining, and the lung injury score was calculated. Quantitative real-time PCR was employed to detect the mRNA expression levels of TNF-α and IL-1β in lung tissues and ELISA was performed to test the protein levels of TNF-α and IL-1β in bronchoalveolar lavage fluid (BALF)...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109141/-hydrogen-rich-saline-attenuates-hyperalgesia-and-reduces-cytokines-in-rats-with-post-herpetic-neuralgia-via-activating-autophagy
#16
Hongtao Ma, Hongguang Chen, Aili Dong, Yanyan Wang, Yingxue Bian, Keliang Xie
Objective To investigate the role of autophagy in hydrogen-rich saline attenuating post-herpetic neuralgia (PHN) in rats. Methods A total of 100 male SD rats were randomly divided into the five groups (n=20): control group, PHN group, PHN group treated with hydrogen-rich saline (PHN-H2 group), PHN group treated with hydrogen-rich saline and 3-MA (PHN-H2-3-MA group), PHN group treated with hydrogen-rich saline and rapamycin (PHN-H2-Rap group). PHN models were established by varicella-zoster virus (VZV) inoculation...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109140/-transplantation-of-bone-marrow-mesenchymal-stem-cell-improves-antioxidant-capacity-and-immune-activity-of-aging-model-rats
#17
Zhihong Wang, Yun Lin, Weimin Chen, Jin Shang, Tiannan Wei
Objective To investigate the impact of bone marrow mesenchymal stem cell (BMSC) transplantation on the antioxidant capacity and immune activity of aging rats induced by D-galactose. Methods Ten healthy male SD rats served as a control group (aged 2 months). To establish aging models, healthy SD rats were daily injected subcutaneously with D-galactose (400 mg/kg). Then the aging model rats were randomized into aging model group and BMSC group (ten rats in each group). The BMSC group was injected with 3×10(6) BMSCs via tail vein...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109139/-imiquimod-combined-with-dendritic-cell-vaccine-decreases-treg-proportion-and-enhances-anti-tumor-responses-in-mice-bearing-melanoma
#18
Shurong Ren, Qiubo Wang, Yanli Zhang, Cuixiu Lu, Ping Li, Yumei Li
Objective To investigate the therapeutic effect of Toll-like receptor 7 (TLR7) agonist imiquimod combined with dendritic cell (DC)-based tumor vaccine on melanoma in mice and the potential mechanism. Methods Melanoma-bearing mouse models were established by subcutanous injection of B16-OVA cells into C57BL/6 mice. DCs were isolated from mouse bone marrow and propagated in culture medium with recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant mouse interleukin-4 (rmIL-4)...
February 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28109079/silencing-of-ribosomal-protein-l34-rpl34-inhibits-the-proliferation-and-invasion-of-esophageal-cancer-cells
#19
Huijie Fan, Jing Li, Yongxu Jia, Jingjing Wu, Long Yuan, Mingjun Li, Jiangqi Wei, Benling Xu
Ribosomal protein L34 (RPL34) belongs to the L34E family of ribosomal proteins and contains a zinc finger motif. Aberrant expression of RPL34 has been reported in several human malignancies. However, the precise role and potential underlying mechanisms of RPL34 in human esophageal cancer remain largely unknown. Thus, the objective of this study was to investigate the role of RPL34 in esophageal cancer progression. Our results showed that the expression of RPL34 at both mRNA and protein levels was frequently up-regulated in esophageal cancer cell lines...
January 20, 2017: Oncology Research
https://www.readbyqxmd.com/read/28109074/determination-of-four-nadolol-stereoisomers-in-capsules-by-high-performance-liquid-chromatography-and-circular-dichroism
#20
Grazielle Prado Alexandre, Márcio Nardelli Wandermuren, Adriano Peron, Vanessa Franco Tavares, Denis Alexandre Ramalho Padeiro, Kelly Alessandra Sakiara, María Segunda Aurora-Prado, Anil Kumar Singh, Erika Rosa Maria Kedor-Hackmann, Maria Inês Rocha Miritello Santoro
Nadolol is a blocking agent with activity in β-adrenergic receptors. The objective of this research was to develop andvalidate an HPLC and circular dichroism method for the quantification of four nadolol stereoisomers in capsules. The HPLC method was validated using a Chiralcel OD column (250 × 4,6 mm, 10 μm). The mobile phase consisted of hexane-ethanol-diethylamine-acetic acid(86 + 14 + 0.3 + 0.3, v/v/v/v), with a flow rate of 0.7 mL/min and temperature of 25 ± 1°C and UV detection carried out at 220 nm...
January 21, 2017: Journal of AOAC International
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