1.Distribution and identification of immunocytes in humanized SCID mouse model
Chen LIN ; Yubo TAN ; Xue BAI ; Shaohua CHEN ; Lijian YANG ; Zhenyou JIANG ; Yangqiu LI
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: Humanized-NOD/SCID(hu-NOD/SCID) mouse model was established and the level of immune reconstitution was assessed in this model. METHODS: Mononuclear cells (MNC) and CD34+ cells were isolated or sorted from cord blood(CB). Human CD45, CD19, CD3 markers on cells from NOD/SCID murine peripheral blood(PB), bone marrow(BM), thymus were detected by FCM from 4 to 10 weeks after hematopoietic stem cell transplantation. After 10 weeks, the gene expressions of the human ?2M and RAG2 were detected by RT-PCR in PB or bone marrow of mice model. RESULTS: Human CD45, CD19, CD3 cells populations in PB and BM were found by flow cytometry in mice model transplanted with CD34+ cells or CB MNC from 4 to 10 weeks. The highest positivity of human lymphocytes was at 8 week after transplantation. The levels of human cell engraftment in mice transplanted with CD34+ cells were higher than those in mice transplanted with CB MNC. The mRNA of human ?2M and RAG2 were found by RT-PCR in BM.CONCLUSION: The higher level of human lymphocyte engraftment is established in NOD/SCID mouse model transplanted with CD34+ compared with CB MNC. The maturation of T lymphocytes could be happened in bone marrow of mice model.
2.Effects of intravenous and topical tranexamic acid on blood loss and inflammatory response after posterior cervical decompressive laminectomy
Chinese Journal of Tissue Engineering Research 2024;28(21):3367-3372
BACKGROUND:With the increase of the incidence of cervical spondylosis,it is particularly important to control the postoperative bleeding and related inflammatory reaction of cervical vertebra. OBJECTIVE:A prospective study was conducted to investigate the combined effect of intravenous and topical tranexamic acid of different concentrations on postoperative blood loss and inflammatory response during posterior cervical decompressive laminectomy combined with lateral mass screw internal fixation and bone graft fusion. METHODS:From January 2020 to December 2022,150 patients who were scheduled to undergo posterior cervical decompressive laminectomy combined with lateral mass screw internal fixation and bone graft fusion for cervical spondylotic myelopathy in Tianjin People's Hospital were enrolled in the study.Patients were divided into A,B,and C groups(n=50)by randomized double-blind lottery.The patients in the A,B,and C groups were treated with 1%,3%,and 5%intravenous and topical tranexamic acid solution,respectively.Postoperative blood loss and inflammation-related indicators were compared among three groups.The occurrence of venous thromboembolism and hematoma was observed after operation. RESULTS AND CONCLUSION:(1)There were significant differences in blood loss-related indexes,such as intraoperative blood loss,overall blood loss,and occult blood loss among the three groups(P<0.01).The above indexes were significantly lower in groups B and C than that in group A.There was no significant difference between groups B and C(P>0.05).(2)On days 1 and 3 after surgery,the levels of inflammatory indicators including serum C-reactive protein and interleukin-6 were increased to varying degrees in the three groups compared to before surgery(P<0.05).As the drug concentration increased,its expression decreased,and there was a significant difference among the three groups(P<0.001).(3)Two cases of intramuscular venous thrombosis appeared in each of the three groups after surgery.No hematoma or pulmonary embolism occurred in all three groups.(4)It is concluded that in posterior cervical decompressive laminectomy combined with lateral mass screw internal fixation and bone graft fusion,intravenous combined with topical application of different concentrations of tranexamic acid is effective in reducing perioperative blood loss and inflammatory response.3%tranexamic acid achieves the same effect without significantly increasing the risk of postoperative venous thromboembolism.
3.Cell-ELA-based determination of binding affinity of DNA aptamer against U87-EGFRvIII cell.
Yan TAN ; Huiyu LIANG ; Xidong WU ; Yubo GAO ; Xingmei ZHANG
Chinese Journal of Biotechnology 2013;29(5):664-671
A15, a DNA aptamer with binding specificity for U87 glioma cells stably overexpressing the epidermal growth factor receptor variant III (U87-EGFRvIII), was generated by cell systematic evolution of ligands by exponential enrichment (cell-SELEX) using a random nucleotide library. Subsequently, we established a cell enzyme-linked assay (cell-ELA) to detect the affinity of A15 compared to an EGFR antibody. We used A15 as a detection probe and cultured U87-EGFRvIII cells as targets. Our data indicate that the equilibrium dissociation constants (K(d)) for A15 were below 100 nmol/L and had similar affinity compared to an EGFR antibody for U87-EGFRvIII. We demonstrated that the cell-ELA was a useful method to determine the equilibrium dissociation constants (K(d)) of aptamers generated by cell-SELEX.
Aptamers, Nucleotide
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metabolism
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Brain Neoplasms
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metabolism
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Cell Line, Tumor
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Glioma
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metabolism
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pathology
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Humans
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Mutation
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Protein Binding
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Receptor, Epidermal Growth Factor
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genetics
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metabolism
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SELEX Aptamer Technique
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methods