1.Effect of GDF11 on expansion of CD8 +memory stem T cells
Xingru MA ; Yingshi CHEN ; Yingtong LIN ; Xu ZHANG ; Haihua LUO ; Chao LIU ; Ting PAN
Chinese Journal of Pathophysiology 2016;32(4):762-768
AIM:To investigate the effect of growth differentiation factor 11 ( GDF11 ) on the expansion of CD8 +memory stem T cells ( Tscm) and to further improve the effect of adoptive immunotherapy.METHODS:Healthy human peripheral blood mononuclear cells ( PBMCs) were isolated by density gradient centrifugation at first.Among the i-solated PBMCs, CD8 +T cells were further purified with MACS microbeads.The CD8 +T cells were then randomly divided into experimental groups and control group.The same volume of different concentrations of GDF11 were added into the ex-perimental groups, and the same volume of PBS solution was added into the control group.Finally, the expansion of Tscm in experimental groups and control group was measured by flow cytometry at several time points.RESULTS:GDF11 sig-nificantly increased the number of Tscm in CD8 +T cells in vitro expansion and also dramatically increased the ratio of Tscm in CD8 +T cells.Furthermore, 400 μg/L GDF11 treatment for 3 weeks was the optimal condition to induce CD8 +Tscm. CONCLUSION:GDF11 effectively increases the number and ratio of Tscm in the CD8 +T cells in cell culture growth, thereby creating a new strategy to further improve the efficiency of adoptive immunotherapy.
2.Inflammatory mechanism and significance of tumor necrosis factor-α and interleukin-1β in pathogenesis of conjunctivochalasis
Meiqing, KE ; Xingru, ZHANG ; Guizhen, ZHOU ; Yanchun, MA ; Qingsong, LI ; Minhong, XIANG
Chinese Journal of Experimental Ophthalmology 2017;35(4):325-331
Background Conjunctivochalasis (CCh) is a common age-related ocular surface diseases.Researches showed that the inflammatory factors are upregulated in conjunctiva and tear of CCh patients,inferring inflammation participates in the pathogenesis of CCh,however,its mechanism is unclear now.Studies also showed that the expression of matrix metalloproteinases (MMPs) in CCh conjunctiva is elevated.However,the association between inflammatory factors and MMPs is unknown.Objective This research was to observe the effects of tumor necrosisfactor-α (TNF-α) and interleukin-1 β (IL-1 β) on the expression of MMP-1,MMP-3,tumor necrosis factor-stimulatedgene-6(TSG-6) and pentraxin-3 (PTX3) in cultured CCh fibroblasts.Methods Conjunctival fibroblasts wereisolated and cultured from CCh-derived and cataract-derived human conjunctiva explants.The cells were treated using20 ng/ml PBS,TNF-α or IL-1β for 4 hours,respectively.The expression levels of MMP-1,MMP-3,TSG-6 and PTX3genes and proteins were detected by quantitative real time PCR and Western blot assay,respectively.Results Thesecond-generation cells showed a long and fusiform shape with ovoid nucleus and radial agreement,and the cellprocessors connected each other.The differences of the relative expression levels of MMP-1,MMP-3,TSG-6 and PTX3mRNA in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-αand IL-1 β (MMP-1 mRNA:Fgroup =2.611,P =0.116;Fi =161.564,P =0.000;MMP-3 mRNA:Fgroup =5.201,P =0.029;Fintervene =211.021,P =0.000;TSG-6 mRNA:Fgroup =47.209,P =0.000;Fintervene =119.340,P =0.000;PTX3 mRNA:Fgroup =40.512,P =0.000;Fintervene =93.935,P =0.000).Compared with the cataract group with PBS treatment,the expression levels of MMP-1,MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with PBS treatment or the cataract group with TNF-α and IL-1 β treatment (all at P<0.05).Compared with the CCh group with PBS treatment,the expression levels of MMP-1,MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with TNF-α and IL-1β treatment (all at P < 0.05).The differences of the relative expression levels of MMP-1,MMP-3,TSG-6 and PTX3 proteins in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-α and IL-1 β (MMP-1:Fgroup =84.702,P =0.000;Fint =48.900,P =0.000;MMP-3:Fgroup =112.818,P =0.000;Fint =194.980,P =0.000;TSG-6:Fgroup =56.867,P =0.000;Fint =70.356,P =0.000;PTX3:Fgroup =1.488,P =0.231;Fint =89.872,P =0.000),and the expression changes of MMP-1,MMP-3,TSG-6 and PTX3 proteins were coincident with the genes among the groups with various treatments.Conclusions The expressions of MMP-1,MMP-3,TSG-6 and PTX3 in conjunctival fibroblasts are upregulated in CCh eyes.The interaction of TNF-α and IL-1 β with MMPs is probably involved with the pathogenesis and development MMPs probably.
3.Associations of polymorphism in large tumor suppressor kinase 2 gene with colorectal cancer
Yingze Li ; Fang Gao ; Xingru Wei ; Wenjie Dong ; Licong Ma ; Tong Dang ; Yanbin Jia
Acta Universitatis Medicinalis Anhui 2022;57(12):1927-1932
Objective :
To investigate the association between single nucleotide polymorphism ( SNP) rs558614, rs9552315,rs7317471 and rs9509492 in large tumor suppressor kinase 2 (LATS2) gene and the risk of colorectal cancer.
Methods :
A total of 390 colorectal cancer patients and 413 healthy subjects were genotyped by Taqman method.The odds ratio ( OR) and its 95% CI were calculated by unconditional logistic regression,to estimate the associations between SNP rs558614,rs9552315,rs7317471,rs9509492 in LATS2 gene and the risk of colorectal cancer,rectal cancer,as well as colon cancer under codominant,dominant,recessive,overdominant,and log-ad- ditive genetic models. Haplotypes were constructed by haploview software 4. 2 .
Results :
SNP rs558614, rs7317471,rs9552315 and rs9509492 in LATS2 gene were not associated with the risk of colorectal cancer,rectal cancer and colon cancer under codominant,dominant,recessive,overdominant,and log-additive genetic models. No haploid blocks were formed between the 4 SNPs.
Conclusion
SNP rs558614 ,rs7317471 ,rs9552315, rs9509492 in LATS2 gene may not play a major role in the development of colorectal cancer,rectal cancer and co- lon cancer.