A brief introduction of identification of tumor antigen-specific TCR gene / 中国免疫学杂志

Adoptive therapy with T cells modified by tumor antigen specific T cell receptor(TCR)gene become a research hotspot in tumor biotherapy.The generation of high-affinity TCR is a significant pre-condition for TCR-T therapy.In this review,we focus on the identification of tumor antigen-specific TCR genes,which based on RT-PCR,single cell RT-PCR and CDR3 length polymorphism analysis.

Application of CRISPR/Cas9 in tumor immunotherapy / 中国免疫学杂志

In recent years,tumor immunotherapy has attracted more and more attention because of its remarkable curative effect and innovation.With the development of CRISPR gene editing system and its application in many fields,researchers have seen its great potential,especially in the field of cancer research.For adoptive cell therapy,CRISPR/Cas9 can knockout endogenous TCR and HLA-Ⅰ genes in TCR-T and CAR-T cells to generate universal effective T cells.It provided a new method to inhibit and block the immune checkpoint.The application of CRISPR/Cas9 in genetic screening makes antibody targeted therapy to a new level.CRISPR has greatly promoted the research of tumor immunotherapy.This article introduced the application of CRISPR/Cas9 gene editing system in tumor immunotherapy.

Effect of miRNA from Glycyrrhiza uralensis decoction on gene expression of human immune cells / 中国中药杂志

MicroRNAs(miRNA) are small non-coding RNAs that regulate the expression of protein coding genes by repressing translation of protein coding mRNA or enhancing mRNA degradation. Its functions have attracted more and more attention from the public. In recent years, the cross-border regulation of miRNA has become a new research direction, and provides a new perspective for people to comprehensively understand the functions of miRNA. Plant miRNA is usually methylated and not easy to degrade. According to our previous researches, there were abundant small RNAs in the decoction of dried liquorice, which provides a new way to study the mechanism of action of licorice. In this study, small RNAs extracted from Glycyrrhiza uralensis decoction and synthesized miRNA mimics were used to treat peripheral blood mononuclear cells(PBMC) isolated from healthy volunteers. The gene expression of toll-like receptors(TLRs), some transcription factors, signal molecules and cytokines were analyzed by RT-PCR. The results showed that glycyrrhiza miRNA could significantly regulate PBMC by inhibiting the expression of genes involved in T cell differentiation, inflammation and apoptosis. The study brought new ideas to us in comprehensively studying the mechanism of licorice and developing the traditional Chinese medicine.

Kinematics and dynamics characteristics of index finger after cold stimulation / 医用生物力学

Objective To investigate effects of cold water stimulation on kinematic and dynamic characteristics of index finger. Method In order to reduce the influence on its flexion and extension, the index finger movement was recorded by a high speed camera. Then a self developed MATLAB program was developed to obtain the trajectory with the help of the black speckles on index finger joints. Finally, a dynamic calculating model with FDS and FDP was set up to describe the relationship between the trajectory and muscle forces changed along with the time. Results After the cold water stimulation, the average angular velocity of the second joint in the index finger decreased in the process of fisting and stretching. Before and after the cold water stimulation, the force of FDP muscle changed little while the force of FDS muscle approximately doubled at the end of the fisting. Conclusions An optical method for obtaining the trajectory of index finger was applied, and a dynamic calculating model was developed to calculate the muscle forces during the finger movement. Therefore, the characteristics of the index finger before and after the cold stimulation could be further discussed.

Tumor-specific T cell recptor gene transfection promotes memory T cell differentiation in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate effect of tumor-specific T cell receptor gene transfection on memory T cell differentiation in vitro.</p><p><b>METHODS</b>TCRVbeta7.1 gene was transferred into peripheral blood mononuclear cells (PBMCs) obtained from healthy adults, and the expression of Vbeta7.1 was detected by flow cytometry before and after the transfection. Memory T cell differentiation was induced by stimulation with the hepatocarcinoma cell line BEL-7402 in vitro. The expression of surface molecules CD45RO, CD45RA and CCR7 was analyzed by flow cytometry to identify the phenotype and subsets of the memory T cells. Fluorescence-activated cell sorting was performed to detect the apoptosis of the tumor cells, and enzyme-linked immunoabsorbent assay was used to determine the production of interferon-gamma (IFN-gamma) for assessing the immune function of the memory T cells.</p><p><b>RESULTS</b>Flow cytometry showed that TCRVbeta7.1 gene was efficiently expressed after transfection. After stimulation by the tumor cells in vitro, the expression of CD45RO in TCRVbeta7.1 gene-modified T cells increased gradually, and analysis of the coexpression of CD45RA and CCR7 revealed that the effector memory T cells constituted the majority of the differentiated memory T cells. The apoptotic rate of the tumor cells induced by the T cells increased significantly with also obviously increased INF-gamma secretion in the memory T cells.</p><p><b>CONCLUSION</b>Tumor-specific TCRVbeta7.1 gene transfection can promote the differentiation of the memory T cells, the majority of which belongs to effector memory T cells that perform immune functions by inducing apoptosis and cytokine secretion.</p>

Effect of Paraformaldehyde Fixation on the FRET Efficiency Calculation between Fluorescent Proteins / 中国生物工程杂志

Objective:To investigate the effect of paraformaldehyde fixation on measuring the protein-protein interaction by fluorescence resonance energy transfer(FRET)to resolve the problem of FRET efficiency calculation in excess-movement cells.Methods:The C terminals of TCR ? chain(TRA)and TCR ? chain(TRB)genes,which were ideal for protein-protein interaction research,were fused with ECFP and EYFP gene respectively by fusion PCR and transferred into target cell.A grou Pcells were fixed in paraformaldehyde(0.5%)for 0.5~1h and another left alive,then these cells were subject to ECFP/EYFP FRET calculation with confocal laser scanning microscope.The ECFP/EYFP FRET efficiencies in live and fixed cell were analyzed and compared.Results:There is no significant statistical difference between the ECFP/EYFP FRET efficiencies of live cell and cell fixed with lower paraformaldehyde concentration and shorter incubation time.Conclusion:fixation with low-concentration paraformaldehyde and short-time incubation has no distinct influence on measuring protein-protein interaction,and facilitated the FRET calculation in excess-movement cells.