Background::B-cell maturation antigen (BCMA)-directed chimeric antigen receptor T (CAR-T) therapy yield remarkable responses in patients with relapsed/refractory multiple myeloma (R/RMM). Circulating tumor DNA (ctDNA) reportedly exhibits distinct advantages in addressing the challenges posed by tumor heterogeneity in the distribution and genetic variations in R/RMM.Methods::Herein, the ctDNA of 108 peripheral blood plasma samples from patients with R/RMM at the First Affiliated Hospital, School of Medicine, Zhejiang University was thoroughly investigated before administration of anti-BCMA CAR-T therapy to establish its predictive potential. Flow cytometry is used primarily to detect subgroups of T cells or CAR-T cells.Results::In this study, several tumor and T cell effector-mediated factors were considered to be related to treatment failure by an integrat analysis, including higher percentages of multiple myeloma (MM) cells in the bone marrow ( P = 0.0125), lower percentages of CAR-T cells in the peripheral blood at peak ( P = 0.0375), and higher percentages of CD8 + T cells ( P = 0.0340). Furthermore, there is a substantial correlation between high ctDNA level (>143 ng/mL) and shorter progression-free survival (PFS) ( P = 0.007). Multivariate Cox regression analysis showed that high levels of ctDNA (>143 ng/mL), MM-driven high-risk mutations (including IGLL5 [ P = 0.004], IRF4 [ P = 0.024], and CREBBP [ P = 0.041]), number of multisite mutations, and resistance-related mutation ( ERBB4, P = 0.040) were independent risk factors for PFS. Conclusion::Finally, a ctDNA-based risk model was built based on the above independent risk factors, which serves as an adjunct non-invasive measure of substantial tumor burden and a prognostic genetic feature that can assist in predicting the response to anti-BCMA CAR-T therapy.

BACKGROUND: B-cell maturation antigen (BCMA)-directed chimeric antigen receptor T (CAR-T) therapy yield remarkable responses in patients with relapsed/refractory multiple myeloma (R/RMM). Circulating tumor DNA (ctDNA) reportedly exhibits distinct advantages in addressing the challenges posed by tumor heterogeneity in the distribution and genetic variations in R/RMM. METHODS: Herein, the ctDNA of 108 peripheral blood plasma samples from patients with R/RMM was thoroughly investigated before administration of anti-BCMA CAR-T therapy to establish its predictive potential. Flow cytometry is used primarily to detect subgroups of T cells or CAR-T cells. RESULTS: In this study, several tumor and T cell effector-mediated factors were considered to be related to treatment failure by an integrat analysis, including higher percentages of multiple myeloma (MM) cells in the bone marrow (P = 0.013), lower percentages of CAR-T cells in the peripheral blood at peak (P = 0.037), and higher percentages of CD8+ T cells (P = 0.034). Furthermore, there is a substantial correlation between high ctDNA level (>143 ng/mL) and shorter progression-free survival (PFS) (P = 0.007). Multivariate Cox regression analysis showed that high levels of ctDNA (>143 ng/mL), MM-driven high-risk mutations (including IGLL5 [P = 0.004], IRF4 [P = 0.024], and CREBBP [P = 0.041]), number of multisite mutations, and resistance-related mutation (ERBB4, P = 0.040) were independent risk factors for PFS. CONCLUSION: Finally, a ctDNA-based risk model was built based on the above independent risk factors, which serves as an adjunct non-invasive measure of substantial tumor burden and a prognostic genetic feature that can assist in predicting the response to anti-BCMA CAR-T therapy. REGISTERATION Chinese Clinical Trial Registry (ChiCTR2100046474) and National Clinical Trial (NCT04670055, NCT05430945).

Objective:To investigate the correlation of antinuclear antibody (ANA) with clinical response to infliximab (IFX) in patients with Crohn′s disease (CD) .Methods:Patients who were diagnosed as CD and treated with IFX in Nanjing Drum Tower Hospital from January 2018 to September 2021 were retrospectively studied. The correlation analysis was used to explore the correlation between ANA and clinical response. These patients were divided into two groups according to the ANA titer after 25 weeks of IFX treatment. The differences in clinical data between the two groups were assessed by univariate analysis. The variables with P<0.15 in univariate analysis and having clinical significance were further analyzed by multivariate Logistic regression to determine the independent risk factors of the induction of ANA. Results:A total of 82 patients with CD were included. Forty-one (50.0%) patients were set as positive group, and 41 (50.0%) patients were set as negative group. In terms of clinical response, the clinical response rates of two groups were 68.3% and 41.5%, and the difference was significant (χ 2 = 5.959, P = 0.015) . Positive group was divided into 1∶100 subgroup ( n = 17) , 1∶320 subgroup ( n = 11) and ≥1∶1000 subgroup ( n = 13) . The clinical response rates of three groups were 41.2%, 45.5% and 7.7% respectively, and the difference was not statistically significant (χ 2 = 5.334, P = 0.084) . The incidences of adverse events in the two groups were 17.1% and 7.3%, and the difference was not significant (χ 2 = 1.822, P = 0.177) . Univariate analysis showed that the difference of total protein (TP) before IFX treatment between the positive group and negative group was statistically significant ( P<0.05) . Multivariate logistic regression analysis showed that age ( OR = 1.060, 95% CI: 1.015 ~ 1.107, P = 0.008) and the baseline TP ( OR = 1.110, 95% CI: 1.023 ~ 1.205, P = 0.012) were the independent risk factors for the induction of ANA. Conclusions:The formation of ANA may affect the clinical response of IFX, so the ANA titer should be monitored regularly during the IFX therapy. In addition, age and baseline TP are related to the formation of ANA.

Objective:To investigate the correlation of antinuclear antibody (ANA) with clinical response to infliximab (IFX) in patients with Crohn′s disease (CD) .Methods:Patients who were diagnosed as CD and treated with IFX in Nanjing Drum Tower Hospital from January 2018 to September 2021 were retrospectively studied. The correlation analysis was used to explore the correlation between ANA and clinical response. These patients were divided into two groups according to the ANA titer after 25 weeks of IFX treatment. The differences in clinical data between the two groups were assessed by univariate analysis. The variables with P<0.15 in univariate analysis and having clinical significance were further analyzed by multivariate Logistic regression to determine the independent risk factors of the induction of ANA. Results:A total of 82 patients with CD were included. Forty-one (50.0%) patients were set as positive group, and 41 (50.0%) patients were set as negative group. In terms of clinical response, the clinical response rates of two groups were 68.3% and 41.5%, and the difference was significant (χ 2 = 5.959, P = 0.015) . Positive group was divided into 1∶100 subgroup ( n = 17) , 1∶320 subgroup ( n = 11) and ≥1∶1000 subgroup ( n = 13) . The clinical response rates of three groups were 41.2%, 45.5% and 7.7% respectively, and the difference was not statistically significant (χ 2 = 5.334, P = 0.084) . The incidences of adverse events in the two groups were 17.1% and 7.3%, and the difference was not significant (χ 2 = 1.822, P = 0.177) . Univariate analysis showed that the difference of total protein (TP) before IFX treatment between the positive group and negative group was statistically significant ( P<0.05) . Multivariate logistic regression analysis showed that age ( OR = 1.060, 95% CI: 1.015 ~ 1.107, P = 0.008) and the baseline TP ( OR = 1.110, 95% CI: 1.023 ~ 1.205, P = 0.012) were the independent risk factors for the induction of ANA. Conclusions:The formation of ANA may affect the clinical response of IFX, so the ANA titer should be monitored regularly during the IFX therapy. In addition, age and baseline TP are related to the formation of ANA.

【Objective】 To analyze the polymorphism of erythrocyte membrane blood group glycoprotein A (GPA) related gene GYPA in high and low endemic population for clonidia sinensis infection, aimed at investigating the correlation between erythrocyte transmembrane glycoproteins and clonorchis sinensis infection. 【Methods】 From Dec 2019 to Jun 2020, anticoagulant blood samples were randomly collected in WuMing district (n=700) and GuiGang district (n=500 ) of Nanning city, and the IgG antibody to clonorchis sinensis in plasma was detected, and the DNA of leukocyte was extracted. The full-length exon and partial intron of GYPA gene were sequenced, mutations were characterized by gene cloning, and the risk of infection was calculated by chi-square test. 【Results】 The yield rate of IgG antibody was 62.7% (439/700) in WuMing district and 3.4% (17/500) in GuiGang district(P<0.05). The insertion of base C at the 54th position of intron-2 in GYPA gene caused the reading frame shift. The mutation was presented in 23.9% (105/439) and 17.6% (3/17) of the population with clonorchis sinensis exposure in WuMing and GuiGang area, respectively, while 49.4% (129/261) and 54.7% (264/483) in the negative population, respectively (P<0.05). 【Conclusion】 The infection rate of clonorchis sinensis in WuMing district was higher than that in GuiGang district. The mutation rate of reading frame shift caused by the insertion of base C at the 54th position of GYPA intron-2 was much lower in the positive population of clonorchis sinensis infection than the negative population, suggesting that the mutation is a protective gene in the negative population of clonorchis sinensis infection. It is necessary to study the mechanism of clonorchis sinensis infection and the mutation point of this gene in order to facilitate the early diagnosis of disease, blood transfusion management, treatment and prevention.

Objective To investigate the effect of gavaged yAGA2-sCT, a recombinant Saccharomyces cerevisiae which expressed salmon calcitonin on its surface, on serum calcium concentration of rats. Methods The sCT gene was artificially synthesized in advance and cloned into the pYD1 vector, a kind of surface displaying plasmid, to yield pYD1-sCT. The recombinant plasmid pYD1-sCT was transformed into S. cerevisiae EBY100 by LiAC method and achieved yAGA2-sCT. The sCT protein, which was expressed on the surface of recombinant S. cerevisiae yAGA2-sCT, was indirectly labeled with FITC. Labeled yeast was immediately detected under the fluorescent microscope. Before being gavaged to rats, recombinant yeast was frozen and vacuum-dried. Lyophilized yeast yAGA2-sCT was administered to rats in dosage of 0.1g/kg, 0.5g/kg and 5.0g/kg, respectively, with 6 rats in each group. Positive drug group (200mU/kg calcitonin via hypodermic injection) and two conditional control groups (being gavaged 5g/kg yAGA2-sCT and 30?g/kg calcitonin, respectively) were acted as controls. The serum was collected from canthus venous plexus and the calcium concentration was immediately measured by automatic biochemical analyzer. Results The sCT protein was successfully expressed on the surface of recombinant yAGA2-sCT, and FITC-labeled yAGA2-sCT was observed under the fluorescent microscope. The recombinant S. cerevisiae yAGA2-sCT was able to reduce the serum calcium concentration of rats in a dose-dependent manner. In the three subgroups, the hypocalcemic effect of high-dosage subgroup, with 5g/kg lyophilized yAGA2-sCT, was the highest (P