Search

Myasthenia gravis (MG) is an autoimmune disease mediated by anti-acetylcholine receptor (AChR) antibodies and dependent on CD4 positive T lymphocytes (CD4+T) cells. MicroRNA (miRNA) is an endogenous small single-stranded non-coding RNA that involved in post-transcriptional regulation and plays an important role in differentiation and function of CD4+T cells. Increasing evidences have shown that changes of miRNAs happened in MG patients, which may participate in the occurrence and development of MG by regulating CD4+T cells, and miRNA may be a potential therapeutic target for MG. In this paper, relationship between miRNA and MG was reviewed in order to provide new ideas for further research on the pathogenesis and clinical diagnosis and treatment of MG. [ABSTRACT FROM AUTHOR]

[Objectives]The paper aimed to investigate the immunological enhancement of acidic Alhagi sparsifolia Shap polysaccharide(aASP)on mouse splenic lymphocytes. [Methods]aASP was extracted by alcohol precipitation, separated and purified by column chromatography, and the contents of sugar, uronic acid, and protein were detected using phenol sulfuric acid method, sulfuric acid-carbazole method, and BCA microplate reader. The polysaccharide properties were identified by infrared spectroscopy. MTT assay was used to determine the safe concentration of aASP on mouse splenic lymphocytes, and the proliferative effects of aASP alone and synergistic lipopolysaccharide(LPS)and phytoheamagglutinin(PHA)on splenic lymphocytes were detected at three concentrations of 500, 125 and 31.25 μg·mL-1 within the safe concentration range; the effect of aASP on lymphocyte differentiation was detected by flow cytometry; the effect of aASP on lymphocyte cytokine secretion was detected by ELISA. [Results]The contents of polysaccharide, uronic acid, and protein in aASP extracts were(81.8±1.2)%, (17.34±0.75)%, and(0.54±0.08)%, respectively. Infrared spectra showed that aASP had a strong absorption peak at 1 746.53 cm-1, and combined with the content of uronic acid, aASP could be identified as acidic polysaccharide. High, medium, and low concentrations of aASP could promote the proliferation of splenic lymphocytes, and aASP synergistic with LPS and PHA could significantly promote the proliferation of B cells and T lymphocytes(P<0.05). The results of flow cytometry and ELISA showed that aASP was able to promote the differentiation of CD4+ and CD8a+ T cells and promote the secretion of IFN-γ, TNF-α, IL-6, and IL-4 in splenic lymphocytes. [Conclusions]aASP can boost the proliferation of splenic lymphocytes in mice, induce Th1(helper T lymphocyte 1)and Th2(helper T lymphocyte 2)mixed immune responses in splenic T lymphocytes, and the immune cells are biased towards CD4+ T cell differentiation. [ABSTRACT FROM AUTHOR]

BACKGROUND: Myeloid-derived suppressor cells are a heterogeneous group of cells derived from bone marrow, which can inhibit the function of T cells and exert immunosuppressive effect. Mesenchymal stem cells are pluripotent stem cells, which can inhibit innate and adaptive immune responses. Myeloid-derived suppressor cells and mesenchymal stem cells have many similarities in immunosuppressive function, which can be related to each other in tumor, inflammation and transplantation. OBJECTIVE: To summarize the mechanisms of action based on myeloid-derived suppressor cells and the effect of mesenchymal stem cells on the negative regulatory effect of immune response in myeloid-derived suppressor cells. METHODS: A computer search of the Web of Science Core Collection database was conducted with key words of “myeloid-derived suppressor cells”, “mesenchymal stem cells”, “stem cells”. Finally, 49 papers were included for review. RESULTS AND CONCLUSION: Myeloid-derived suppressor cells play an important immunosuppressive role in vivo by secreting arginase-1, inducible nitric oxide synthase, and indoleamine 2,3-dioxygenase. Myeloid-derived suppressor cells and mesenchymal stem cells interact through different mechanisms in the treatment of autoimmune diseases, organ transplantation, inflammatory infections, tumors and other diseases, causing tumor cells to evade immune surveillance and promoting further tumor progression. In non-tumor diseases, myeloid-derived suppressor cells play a certain benign regulatory role, weakening the body’s own immune response and improving clinical symptoms. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to construct the eukaryotic expression vector of rotavirus（Rotavirus, RV）non-structural protein1（NSP1）and 3（NSP3）and express it in human embryonic kidney epithelial HEK239 T cells, and then to study the preliminary effect of rotavirus NSP3 on the activity of IFN-β/ISRE reporter gene in vitro. [Method] First, the NSP1 and NSP3 coding genes of ape rotavirus SA11 strain were synthesized and cloned into the cloning vector pUCm-T, the target fragments of NSP1 and NSP3 were amplified by PCR, and then the eukaryotic expression plasmids specifically expressing recombinant proteins pRK-Flag-NSP1/pRK-HA-NSP1 and pRK-Flag-NSP3/pRK-HA-NSP3 were constructed by using eukaryotic expression vectors pRK-Flag and pRK-HA as skeletons. The plasmids were transfected into HEK293 T cells after restriction endonuclease digestion and sequencing. The expression of NSP1/NSP3 protein was identified by Western blot. Then, the recombinant plasmid pRK-Flag-NSP1/pRK-Flag-NSP3 and IFN-β-Luc/ISRE-Luc plasmid were co-transfected into HEK239 T cells. After stimulated by RIG-IN（RIG-I activation domain）, the effect of NSP1/NSP3 protein on the activity of IFN-β/ISRE reporter gene was determined by double luciferase reporter gene system. [Result] The full-length genes of NSP1 and NSP3 were cloned successfully, and the eukaryotic expression plasmids pRK-Flag-NSP1/pRK-HA-NSP1 and pRK-Flag-NSP3/pRK-HA-NSP3, of NSP1 and NSP3 protein of ape rotavirus strain SA11 were constructed. The successful expression of NSP1 and NSP3 protein was confirmed by transfection and expression of Western blot in HEK293 T cells. Recombinant plasmids pRK-Flag-NSP1 and pRK-Flag-NSP3 could significantly reduce the activity of double luciferase induced by RIG-IN. [Conclusion] NSP1 and NSP3 proteins can inhibit the activities of IFN-β and ISRE promoters stimulated by RIG-IN, which proves that NSP3 protein has the same function as NSP1 protein in inhibiting IFN-β signal pathway, which lays a theoretical foundation for further study on the mechanism of NSP3 protein regulating type I interferon signal pathway. [ABSTRACT FROM AUTHOR]

Objective: To investigate clinical efficacy of pediatric massage combined with Xiaoer Xiaoji Zhike oral liquid and azithromycin in the treatment of children of mycoplasma pneumoniae pneumonia (MPP) and its effect on immune function and inflammatory factors. Methods: 80 cases of children with MPP who were treated in our hospital from March 2018 to June 2019 were selected. According to the different treatment methods, they were divided into the control group (azithromycin and Xiaoer Xiaoji Zhike oral liquid treatment, n=40) and the study group (pediatric massage, Xiaoer Xiaoji Zhike oral liquid, azithromycin treatment, n=40). The clinical efficacy of the two groups and the improvement of children's symptoms during the treatment were observed, peripheral blood T lymphocyte subsets, adverse reactions and serum inflammatory factors between the two groups were compared. Results: Compared with the control group, the clinical total effective rate of the study group was higher (P<0.05). Compared with the control group, the study group had a shorter of lung rales, cough and X-ray film of lung lesions to disappear time and antipyretic time (P<0.05). Compared with the control group, CD3+, CD4+ and CD4+/CD8+ in the study group was higher at 14 d after treatment, CD8+ was lower (P<0.05). Compared with the control group, interleukin-1β (IL-1β), C-reactive protein (CRP) and interleukin-6 (IL-6) in the study group were lower (P<0.05). There was no significant difference in incidence of adverse reactions in two groups (P>0.05). Conclusion: Pediatric massage combined with Xiaoer Xiaoji Zhike oral liquid and azithromycin in the treatment of children with MPP can promote the improvement of symptoms, optimize the clinical treatment effect, and effectively regulate the immune function and inflammatory factor levels of children. [ABSTRACT FROM AUTHOR]

【Objective】This paper aimed to establish the 293 T and HT 1080 cell lines with CRISPR/Cpf1 gene by lentiviral vector and further to realize the high-efficiency gene editing. 【Method】AsCpf1 and LbCpf1 gene was amplified by PCR technique, which was inserted into the pLent-EF1 a-MF-CMV-GFP-P2 A-Puro vector. Monolayer of 293 FT cells were cotransfected with three plasmids psPAX2, pMD2.G and Cpf1 in pLent-EF1 a-MF-CMV-GFP-P2 A-Puro. The recombinant lentivirus expressing LbCpf1 and AsCpf1 was harvested, and the titer of virus was calculated by fluorescence microscope 48 hours later. The purified lentivirus was used to infect 293 T cell and the transduced cells were selected with puromycin and single cell colonies were isolated and verified for DNA, RNA and cDNA. Single cell colonies were transfected by sgRNA targeting the exons ABCG1 CDS regions to the functional domains of ABCG1 genes, and gene knockout efficiency was measured by DNA specificity and T7 EΙ digestion. 【Result】DNA sequnencing verified that Cpf1-pLent was constructed successfully, and the titer of recombinant lentivirus was greater than 10~6 TU/mL. Three cell lines expressing Cpf1 stably were obtained, two of which(AsCpf1-293 T-7 D and AsCpf1-HT1080 02-3 B) the targeted gene was knocked out successfully by T7 EΙ digestion. 【Conclusion】 293 T and HT 1080 cells stably expressing Cpf1 was successfully constructed, which could be a valuable tool for gene editing. [ABSTRACT FROM AUTHOR]