Objective: To evaluate the safety and efficacy of therapeutic whole blood exchange combined with lymphoplasmapheresis in the treatment of refractory autoimmune hemolytic anemia (AIHA). Methods: A retrospective analysis was performed on the clinical data of AIHA patients who underwent therapeutic whole blood exchange combined with lymphoplasmapheresis at our hospital from March 2022 to May 2025. Efficacy was assessed by comparing changes in hemoglobin, platelet count, and bilirubin levels before and after treatment. Safety was evaluated by analyzing vital signs before and after the procedure, parameters during the exchange, and adverse reactions. Results: A total of 12 AIHA patients were enrolled, completing 19 exchange procedures. The number of procedures per patient ranged from 1 to 3. The median treatment duration was 67 (65-73) minutes, with a median exchange volume of 2 025 (1 851-2 121) mL, comprising 4.5 (4-6) units of red blood cells and 1 350 (1 200-1 400) mL of plasma. Ten patients achieved partial remission, one achieved complete remission, and one showed no response, yielding an response rate of 91% (11/12). After a single session, hemoglobin increased significantly by 17.58±9.85 g/L (P<0.01), while platelets counts decreased by 45 (17.5, 79)×10 /L (P<0.05), and both systolic and diastolic blood pressure showed a significant elevation (P<0.05). However, no statistically significant differences were observed in total bilirubin, indirect bilirubin, white blood cell count, or heart rate. During the procedures, 4 adverse reactions occurred in 3 patients: one child experienced severe heart rate fluctuation twice consecutively, and two adults developed plasma allergies. All reactions resolved spontaneously without pharmacological intervention. Conclusion: The combination of therapeutic whole blood exchange and lymphoplasmapheresis appears to be a safe and effective treatment for refractory AIHA patients.

The liver is one of the important metabolic organs in the body, and liver diseases are also one of the major public health issues globally. Currently, the lack of models that accurately simulating human physiology and pathology is the key challenge in liver disease research, and the emerging of organoid technology is an excellent solution to this challenge. Organoids are three-dimensional cellular structures cultured in vitro that can mimic the structure and function of real organs while maintaining genetic stability. The development of organoids has provided possibilities for personalized medicine and precision therapy and has become a key tool in the research and treatment of liver diseases. The authors focuse on the application and development of organoids in liver disease research, while also emphasizing their limitations and future prospects.

AIM:This study aims to investigate of perodic expression,distribution and interaction between es-trogen receptor α(ERα)and ClC-3 chloride channel,and their relevance to the cell cycle specificity of tamoxifen(TAM)in anti-breast cancer treatment.METHODS:We utilized a web database to analyze the correlation between ERα and ClC-3 expression.Three-dimentional molecular simulation software and co-immunoprecipitation were employed to detect and analyze the interactions between these two proteins.To assess cell cycle dynamics,we performed thymidine(TdR)double-blocking release assay and used nocodazole to block the cell cycle,with subsequent analysis via flow cytometry.Cell viability was measured by MTT assay.Western blot was conducted to evaluate the protein expression levels of ERα and ClC-3,while immunofluorescence staining was utilized to assess their subcellular distribution.RESULTS:(1)Anal-ysis from the web database revealed a significant correlation between ERα and ClC-3 expression,and co-immunoprecipita-tion confirmed their interaction.(2)We successfully obtained human breast cancer T47D cells at different cycle stages us-ing the TdR double-blocking release method and nocodazole treatment.(3)Treatment with TAM primarily inhibited T47D cell viability during G2/M phase.(4)Both ERα and ClC-3 exhibited cyclic variations in protein expression,with their sub-cellular distributions showing periodicity and co-localization.(5)Protein interactions between ERα and ClC-3 were ob-served across all cell cycle phases;(6)After TAM treatment,ERα expression peaked in G2/M phase,while ClC-3 expres-sion remained unaffected.CONCLUSION:Our findings demonstrate cyclic differences in the expression and distribution of ERα and ClC-3 in human breast cancer T47D cells,along with confirmed interactions between these two proteins.The cyclic properties of ERα may play a role in mediating the cell cycle specificity of TAM's anti-breast cancer effect.

The liver is one of the important metabolic organs in the body, and liver diseases are also one of the major public health issues globally. Currently, the lack of models that accurately simulating human physiology and pathology is the key challenge in liver disease research, and the emerging of organoid technology is an excellent solution to this challenge. Organoids are three-dimensional cellular structures cultured in vitro that can mimic the structure and function of real organs while maintaining genetic stability. The development of organoids has provided possibilities for personalized medicine and precision therapy and has become a key tool in the research and treatment of liver diseases. The authors focuse on the application and development of organoids in liver disease research, while also emphasizing their limitations and future prospects.

AIM:This study aims to investigate of perodic expression,distribution and interaction between es-trogen receptor α(ERα)and ClC-3 chloride channel,and their relevance to the cell cycle specificity of tamoxifen(TAM)in anti-breast cancer treatment.METHODS:We utilized a web database to analyze the correlation between ERα and ClC-3 expression.Three-dimentional molecular simulation software and co-immunoprecipitation were employed to detect and analyze the interactions between these two proteins.To assess cell cycle dynamics,we performed thymidine(TdR)double-blocking release assay and used nocodazole to block the cell cycle,with subsequent analysis via flow cytometry.Cell viability was measured by MTT assay.Western blot was conducted to evaluate the protein expression levels of ERα and ClC-3,while immunofluorescence staining was utilized to assess their subcellular distribution.RESULTS:(1)Anal-ysis from the web database revealed a significant correlation between ERα and ClC-3 expression,and co-immunoprecipita-tion confirmed their interaction.(2)We successfully obtained human breast cancer T47D cells at different cycle stages us-ing the TdR double-blocking release method and nocodazole treatment.(3)Treatment with TAM primarily inhibited T47D cell viability during G2/M phase.(4)Both ERα and ClC-3 exhibited cyclic variations in protein expression,with their sub-cellular distributions showing periodicity and co-localization.(5)Protein interactions between ERα and ClC-3 were ob-served across all cell cycle phases;(6)After TAM treatment,ERα expression peaked in G2/M phase,while ClC-3 expres-sion remained unaffected.CONCLUSION:Our findings demonstrate cyclic differences in the expression and distribution of ERα and ClC-3 in human breast cancer T47D cells,along with confirmed interactions between these two proteins.The cyclic properties of ERα may play a role in mediating the cell cycle specificity of TAM's anti-breast cancer effect.

Objective:To investigate the effect and mechanism of human umbilical cord mesenchymal stem cells (hUCMSCs) transplantation on testicular function in aging mice with oxidative damage.Methods:Totally 18 SPF grade C57BL/C male mice aged 6-8 weeks were randomly divided into 3 groups using a complete randomization method. In control group, mice were injected with an equal amount of physiological saline; in model group, mice were subcutaneous injected D-galactose into the neck and back for 9 consecutive weeks, on the 4th weekend of modeling, the mice were injected with physiological saline via the tail vein; in hUCMSCs group: mice were subcutaneous injected D-galactose into the neck and back for 9 consecutive weeks, on the 4th weekend of modeling, the mice were injected with hUCMSCs via the tail vein of each mouse. After 9 weeks, body weight and testicular weight of the three groups mice were measured and testicular index was calculated. The contents of testosterone, malondialdehyde (MDA) and superoxide dismutase (SOD) in serum and testicular tissue were detected by enzyme-linked immunosorbent assay (ELISA) method. Visual observation of testicular appearance, the histopathological changes of testis were observed by hematoxylin-eosin (HE) staining, and the expression of NF-E2-related factor 2 (Nrf2) signal transduction-related genes and proteins were detected by RT-PCR and Western blotting, respectively.Results:Compared with control group [(0.81±0.13)%], the testicular index of mice in model group [(0.64±0.05)%, P=0.006] was decreased. In model group, the volume of testis was reduced, the integrity of spermatogenic tubules was damaged, spermatogenic cells and sperm were reduced, and the interstitium was sparse. In model group, serum testosterone [(4.10±0.67) μg/L] and SOD [(48.87±6.40) U/mg Prot] were decreased compared with control group [(5.71±0.81) μg/L, P=0.002; (78.53±9.70) U/mg Prot, P=0.001], MDA [(1.11±0.19) nmol/mg Prot] was increased compared with control group [(0.77±0.07) nmol/mg Prot, P=0.001], Keap1 mRNA and protein expression were increased ( P=0.006, P=0.043). The expression levels of Nrf2, SOD and NQO1 mRNA were significantly lower than those in control group ( P=0.002, P<0.001, P=0.001), and the expression levels of Nrf2 and HO-1 protein were significantly lower than those in control group ( P=0.011, P=0.021). Compared with the model group, the testicular index [(0.79±0.03)%, P=0.010] increased in hUCMSCs group, and the tissue structure of testis was clear and complete, spermatogenic cells at all levels of spermatogenic tubules, spermatogenic cells and stromal cells were abundant. Compared with the model group, the content of dihydrotestosterone [(5.24±0.21) μg/L, P=0.028] in serum and SOD [(79.47±14.32) U/mg Prot, P=0.001] in testicular tissue increased in hUCMSCs group, while the content of MDA [(0.77±0.08) nmol/mg Prot, P=0.001] decreased, Nrf2, SOD and NQO1 mRNA expression levels increased ( P=0.024, P=0.037, P=0.005), Keap1 mRNA expression decreased ( P=0.044), Nrf2 and HO-1 proteins expression increased ( P=0.009, P=0.012), while Keap1 protein expression decreased ( P=0.035). There were no statistically significant differences in testicular index, serum testosterone, SOD and MDA between hUCMSCs group and control group (all P>0.05). Conclusion:hUCMSCs significantly improve testicular structure and function damage caused by oxidative damage in aging mice, and the mechanism of action is related to upregulating Nrf2 signaling and downstream antioxidant activity SOD and HO-1 protein expression, reducing Keap1 mediated Nrf2 degradation.

Objective:To investigate the proliferation kinetics of T cells in patients with B-cell hematologic malignancies who received CAR19 T cell therapy.Methods:Observational study. Flow cytometry was used to monitor the levels of CAR19+and CAR19-T cell expansion and the dynamic changes of T lymphocyte subsets before and after CAR19 T cell therapy. The 52 patients with B-cell hematologic malignancies (including 12 B-ALL and 40 NHL) who received CAR19 T cell therapy in the First Affiliated Hospital of Soochow University from November 2021 to December 2023 were recruited in this study. Patients were divided into complete response group and incomplete response group according to the efficacy evaluation criteria in the treatment guidelines for B-cell hematologic malignancies. T test or non-parametric rank sum test were used to compare the differences of CAR19+and CAR19-T cell subsets between the two groups.Results:At the peak of CAR19+T cell expansion, there was no statistic difference of CAR19+T cell subsets between the complete response group and the incomplete response group. After 6 months, the percentage of CD4+T cells (CD3+CD4+CD8-) in CAR19-T cells in patients was lower than the pre-treatment level(48.0+27.2,63.1+19.7,<0.01), and the percentages of CD197+CD45RA+and CD197-CD45RA-subsets recovered to the pre-treatment level, while the percentage of CD197-CD45RA+subset(4.2+3.0,21.1+15.6,<0.01) was lower than the pre-treatment level. The percentage of CD8+T cells (CD3+CD4-CD8+) returned to pre-treatment level after 6 months, CD197-CD45RA-subset in CD8+T cells returned to pre-treatment level, while CD197+CD45RA+subset(16.6+8.7,35.1+30.1,<0.01),CD197+CD45RA-subset(18.7+9.1,25.8+19.1,<0.01) were still lower than pre-treatment level.Conclusion:After CAR19 T cell treatment, there was no significant differences in the proportions of CAR19+T cell subsets in patients with different therapeutic effects. After treatment, the proportion of CAR19-CD3+CD4-CD8+cells recovered earlier than CD3+CD4+CD8-cells, and the dynamic changes of each subgroup were different. This therapeutic regimen has a great impact on the subpopulation of CAR19-T cells in vivo, and the reconstruction of such T cells takes a long time.

Objective:To investigate the effect and mechanism of human umbilical cord mesenchymal stem cells (hUCMSCs) transplantation on testicular function in aging mice with oxidative damage.Methods:Totally 18 SPF grade C57BL/C male mice aged 6-8 weeks were randomly divided into 3 groups using a complete randomization method. In control group, mice were injected with an equal amount of physiological saline; in model group, mice were subcutaneous injected D-galactose into the neck and back for 9 consecutive weeks, on the 4th weekend of modeling, the mice were injected with physiological saline via the tail vein; in hUCMSCs group: mice were subcutaneous injected D-galactose into the neck and back for 9 consecutive weeks, on the 4th weekend of modeling, the mice were injected with hUCMSCs via the tail vein of each mouse. After 9 weeks, body weight and testicular weight of the three groups mice were measured and testicular index was calculated. The contents of testosterone, malondialdehyde (MDA) and superoxide dismutase (SOD) in serum and testicular tissue were detected by enzyme-linked immunosorbent assay (ELISA) method. Visual observation of testicular appearance, the histopathological changes of testis were observed by hematoxylin-eosin (HE) staining, and the expression of NF-E2-related factor 2 (Nrf2) signal transduction-related genes and proteins were detected by RT-PCR and Western blotting, respectively.Results:Compared with control group [(0.81±0.13)%], the testicular index of mice in model group [(0.64±0.05)%, P=0.006] was decreased. In model group, the volume of testis was reduced, the integrity of spermatogenic tubules was damaged, spermatogenic cells and sperm were reduced, and the interstitium was sparse. In model group, serum testosterone [(4.10±0.67) μg/L] and SOD [(48.87±6.40) U/mg Prot] were decreased compared with control group [(5.71±0.81) μg/L, P=0.002; (78.53±9.70) U/mg Prot, P=0.001], MDA [(1.11±0.19) nmol/mg Prot] was increased compared with control group [(0.77±0.07) nmol/mg Prot, P=0.001], Keap1 mRNA and protein expression were increased ( P=0.006, P=0.043). The expression levels of Nrf2, SOD and NQO1 mRNA were significantly lower than those in control group ( P=0.002, P<0.001, P=0.001), and the expression levels of Nrf2 and HO-1 protein were significantly lower than those in control group ( P=0.011, P=0.021). Compared with the model group, the testicular index [(0.79±0.03)%, P=0.010] increased in hUCMSCs group, and the tissue structure of testis was clear and complete, spermatogenic cells at all levels of spermatogenic tubules, spermatogenic cells and stromal cells were abundant. Compared with the model group, the content of dihydrotestosterone [(5.24±0.21) μg/L, P=0.028] in serum and SOD [(79.47±14.32) U/mg Prot, P=0.001] in testicular tissue increased in hUCMSCs group, while the content of MDA [(0.77±0.08) nmol/mg Prot, P=0.001] decreased, Nrf2, SOD and NQO1 mRNA expression levels increased ( P=0.024, P=0.037, P=0.005), Keap1 mRNA expression decreased ( P=0.044), Nrf2 and HO-1 proteins expression increased ( P=0.009, P=0.012), while Keap1 protein expression decreased ( P=0.035). There were no statistically significant differences in testicular index, serum testosterone, SOD and MDA between hUCMSCs group and control group (all P>0.05). Conclusion:hUCMSCs significantly improve testicular structure and function damage caused by oxidative damage in aging mice, and the mechanism of action is related to upregulating Nrf2 signaling and downstream antioxidant activity SOD and HO-1 protein expression, reducing Keap1 mediated Nrf2 degradation.

Objective:To investigate the proliferation kinetics of T cells in patients with B-cell hematologic malignancies who received CAR19 T cell therapy.Methods:Observational study. Flow cytometry was used to monitor the levels of CAR19+and CAR19-T cell expansion and the dynamic changes of T lymphocyte subsets before and after CAR19 T cell therapy. The 52 patients with B-cell hematologic malignancies (including 12 B-ALL and 40 NHL) who received CAR19 T cell therapy in the First Affiliated Hospital of Soochow University from November 2021 to December 2023 were recruited in this study. Patients were divided into complete response group and incomplete response group according to the efficacy evaluation criteria in the treatment guidelines for B-cell hematologic malignancies. T test or non-parametric rank sum test were used to compare the differences of CAR19+and CAR19-T cell subsets between the two groups.Results:At the peak of CAR19+T cell expansion, there was no statistic difference of CAR19+T cell subsets between the complete response group and the incomplete response group. After 6 months, the percentage of CD4+T cells (CD3+CD4+CD8-) in CAR19-T cells in patients was lower than the pre-treatment level(48.0+27.2,63.1+19.7,<0.01), and the percentages of CD197+CD45RA+and CD197-CD45RA-subsets recovered to the pre-treatment level, while the percentage of CD197-CD45RA+subset(4.2+3.0,21.1+15.6,<0.01) was lower than the pre-treatment level. The percentage of CD8+T cells (CD3+CD4-CD8+) returned to pre-treatment level after 6 months, CD197-CD45RA-subset in CD8+T cells returned to pre-treatment level, while CD197+CD45RA+subset(16.6+8.7,35.1+30.1,<0.01),CD197+CD45RA-subset(18.7+9.1,25.8+19.1,<0.01) were still lower than pre-treatment level.Conclusion:After CAR19 T cell treatment, there was no significant differences in the proportions of CAR19+T cell subsets in patients with different therapeutic effects. After treatment, the proportion of CAR19-CD3+CD4-CD8+cells recovered earlier than CD3+CD4+CD8-cells, and the dynamic changes of each subgroup were different. This therapeutic regimen has a great impact on the subpopulation of CAR19-T cells in vivo, and the reconstruction of such T cells takes a long time.