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CD19-targeted chimeric antigen receptor T lymphocytes (CAR-T) has demonstrated a high proportion of complete remission in the treatment of relapsed refractory acute B cell lymphoblastic leukemia (r/r B-ALL). It is of great clinical significance to explore which factors will impact long-term disease-free survival of patients with r/r B-ALL after CAR-T therapy without bridging bone marrow transplantation. Our study found that, in patients with r/r B-ALL without bridging transplantation, the patients' age; infusion dosage; whether they had undergone allo-stem cell transplantation before CAR-T therapy, using CD-19-targeted or CD19/CD22-dual-targeted CAR-T; whether there is fusion gene; tumor burden before therapy; and comorbidity had no significant relationship with their long-term disease-free survival. We found only that CAR-T persistence was highly correlated with patients' long-term disease-free survival. So, we further profiled CAR-T cells using single-cell sequencing and found that there is a specific T cell subset that may be associated with the long-term persistence of CAR-T. Finally, according to the single-cell sequencing results, we established cell production process named PrimeCAR, which shared common signaling pathways with the T cell subset identified. In the preliminary clinical study, prime CAR-Ts yield good persistence in peripheral blood of patients with B-ALL and lymphoma, without observing grade 2 or higher cytokine release syndrome.
ABSTRACT
OBJECTIVE:To study the inhibitory effect and mechanism of Inula helenium ethyl acetate extract(IHE)on prolif-eration of human pancreatic cancer Capan-2 cells. METHODS:MTT was used to determine the cell proliferation inhibition rate af-ter treated by 0,0.5,1,2,4,8 μg/mL IHE;clone formation test was used to observe the effects of 0,1,2 μg/mL IHE treating for 1 week on cell clone formation;Hoechest 33342 staining was used to observe the changes of nuclear morphology after treated by 0,2,4 μg/mL IHE for 48 h;flow cytometry was used to detect the cell apoptosis rate after treated by 0,4,8,16 μg/mL IHE for 48 h;JC-1 staining was used to observe the changes of intracellular mitochondrial membrane potential after treated by 0,4,8, 16 μg/mL IHE for 24 h;Western blot was used to detect the expressions of mitochondrial apoptosis-related proteins Bcl-2,Bax, Mcl-1,p53 up-regulated modulator of apoptosis (PUMA),and polymerase (PARP) after treated by 0,4,8,16 μg/mL IHE for 48 h. RESULTS:2,4,8 μg/mL IHE had obvious inhibitory effect on cell proliferation,showing concentration-dependent relation-ship,with IC50 of 6.6 μg/mL;1,2 μg/mL IHE can obviously inhibit the clone formation of cells;4 μg/mL IHE can obviously cause cell nuclear condensation;8,16 μg/mL IHE can obviously promote the cell apoptosis,and the cell apoptosis rate reached 45.53% after treated by 16 μg/mL IHE for 48 h;16 μg/mL IHE treating for 24 h can cause the decrease of 82.47% cells'mito-chondrial membrane potential;8 μg/mL IHE can obviously down-regulate the protein expressions of Bcl-2,Mcl-1,PUMA and PARP,and 16 μg/mL IHE can obviously down-regulate the expressions of Mcl-1 and PUMA. CONCLUSIONS:IHE may show its inhibitory effect on proliferation of human pancreatic cancer Capan-2 cells by causing the decrease of mitochondrial mem-brane potential in cells and down-regulating the protein expres-sions of Mcl-1 and PUMA to cause cell apoptosis.
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Background Replase of uveitis is a primary cause of vision damage.To predict recurrentassociated factors for uveitis is very critical for the prevention and management of uveitis.Objective This study was to explore the risk factors of recurrent uveitis and establish the prediction model of recurrent uveitis.Methods Clinical data of recurrent uveitis patients who were diagnosed in Renhe Hospital of Three Gorges University from July 1,2010 to June 30,2011 were retrospectively reviewed.The demography characteristics of the patients were collected and the disease was followed-up under the informed consent.Kaplan-Meier method was used to estimate the disease recurrence rate and to plot relapse-free survival curve at different levels of predictive factors.Multivariate Cox proportional hazards model was used to select independent risk factors of relapse and establish the prediction model for recurrent uveitis.Results Total 825 cases of recurrent uveitis were included and followed up for 1 month to 38 months,with a median following-up time of 16 months.Relapse of uveitis was identified in 149 cases (18.1％)during the following-up duration.The relapse-free survival time was from 1 month to 38 months,and the 1-,2-and 3-year cumulative recurrence-free survival rates were 87.3％,82.8％ and 80.9％.Multivariate Cox proportional hazards regression analysis showed that immunosuppression withdrawal(X1) (β =0.940,Waldx2 =12.018,P =0.001),oral steroid withdrawal (X2) (β =1.334,Wald x2 =18.450,P ＜ 0.001),colds (X3) (β =0.642,Wald x2 =11.988,P =0.001),work and study stress(X4) (β=0.285,Wald x2 =4.925,P=0.026) and excessive alcohol and tobacco(X5) (3--0.541,Wald x2 =4.718,P =0.030) were the independent risk factors for recurrence of uveitis.The risk of recurrence in patients with uveitis function model expression was h (t)=h0 exp (2.559 X1 +3.797 X2 + 1.901 X3 + 1.331 X4 +1.719 X5).Conclusions Replase of uveitis is an interaction of many factors,and immunosuppression withdrawal,oral steroid withdrawal,colds,work and study stress,excessive alcohol and tobacco are independent risk factors for recurrence of uveitis.An intervention according to the controllable factors is one of the important ways to prevent the recurrence of uveitis.
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Life science is based on an experimental basis, so the basic experimental teaching for biological and medical graduates is very important. Targeted basic experimental teaching content can help biological and medical graduates lay a solid experiment foundation and promote them to fol-low the development of cutting-edge bioscience.The basic experimental teaching content for biological graduate is set up into four experimental categories uniformly: molecular biology, proteomics experi-ments, bioinformatics experiments and large equipment use. Each category is further set into three experimental projects: validation experiments, comprehensive experiments and independent research experiment. By increasing the degree of openness of large instruments and equipment, improving teaching methods and perfecting appraisal methods, ggraduates' experimental skills, research ability and innovation ability are trained, and the quality of experimental teaching has been improved.