Objective:This study aimed to analyze cytomegalovirus (CMV) reactivation and its influencing factors in patients with B-lymphocyte malignancy who received chimeric antigen receptor T (CAR-T) cell therapy.Methods:This study retrospectively reviewed patients with B-lymphocyte malignancy who received CAR-T cell therapy in the First Affiliated Hospital of Soochow University from January 2021 to December 2023. The data of patients who underwent CMV-DNA detection and/or pathogen metagenomic sequencing twice or more within 100 days after CAR-T cell therapy were analyzed. The clinical characteristics of the CMV reactivation and non-activation groups were compared. The factors related to CMV reactivation were analyzed with the Chi-square test and nonparametric rank sum test, and the risk factors were examined with Logistic regression.Results:This study included 86 patients, among whom 18 (20.9%) had CMV reactivation, and the median time of reactivation was 20 (1-95) days. All of the 18 patients had CMV viremia, and no CMV disease was observed. Seven patients turned to the latent state after continuing acyclovir antiviral therapy, and 11 patients returned to the latent state after upgrading the antiviral therapy to first-line drugs, including ganciclovir and foscarnet sodium. Six or more courses of anti-tumor treatment before CAR-T cell therapy, allogeneic hematopoietic stem cell transplantation within 2 years before CAR-T cell therapy, non-remission before treatment, and the use of high-dose glucocorticoids and/or tocilizumab were related to CMV reactivation, among which allogeneic hematopoietic stem cell transplantation within 2 years pre-treatment and the use of high-dose glucocorticoids and/or tocilizumab treatment were independent risk factors for CMV reactivation.Conclusion:Patients with B-lymphocyte malignancy who received CAR-T cell therapy have the risk of CMV reactivation, especially for those who received allogeneic hematopoietic stem cell transplantation within 2 years pre-treatment and those who received high-dose glucocorticoids and/or tocilizumab treatment.

Objective:This study aimed to analyze cytomegalovirus (CMV) reactivation and its influencing factors in patients with B-lymphocyte malignancy who received chimeric antigen receptor T (CAR-T) cell therapy.Methods:This study retrospectively reviewed patients with B-lymphocyte malignancy who received CAR-T cell therapy in the First Affiliated Hospital of Soochow University from January 2021 to December 2023. The data of patients who underwent CMV-DNA detection and/or pathogen metagenomic sequencing twice or more within 100 days after CAR-T cell therapy were analyzed. The clinical characteristics of the CMV reactivation and non-activation groups were compared. The factors related to CMV reactivation were analyzed with the Chi-square test and nonparametric rank sum test, and the risk factors were examined with Logistic regression.Results:This study included 86 patients, among whom 18 (20.9%) had CMV reactivation, and the median time of reactivation was 20 (1-95) days. All of the 18 patients had CMV viremia, and no CMV disease was observed. Seven patients turned to the latent state after continuing acyclovir antiviral therapy, and 11 patients returned to the latent state after upgrading the antiviral therapy to first-line drugs, including ganciclovir and foscarnet sodium. Six or more courses of anti-tumor treatment before CAR-T cell therapy, allogeneic hematopoietic stem cell transplantation within 2 years before CAR-T cell therapy, non-remission before treatment, and the use of high-dose glucocorticoids and/or tocilizumab were related to CMV reactivation, among which allogeneic hematopoietic stem cell transplantation within 2 years pre-treatment and the use of high-dose glucocorticoids and/or tocilizumab treatment were independent risk factors for CMV reactivation.Conclusion:Patients with B-lymphocyte malignancy who received CAR-T cell therapy have the risk of CMV reactivation, especially for those who received allogeneic hematopoietic stem cell transplantation within 2 years pre-treatment and those who received high-dose glucocorticoids and/or tocilizumab treatment.

Aluminum adjuvants (Alum), approved by the US Food and Drug Administration, have been extensively used in vaccines containing recombinant antigens, subunits of pathogens, or toxins for almost a century. While Alums typically elicit strong humoral immune responses, their ability to induce cellular and mucosal immunity is limited. As an alternative, layered double hydroxide (LDH), a widely used antacid, has emerged as a novel class of potent nano-aluminum adjuvants (NanoAlum), demonstrating advantageous physicochemical properties, biocompatibility and adjuvanticity in both humoral and cellular immune responses. In this review, we summarize and compare the advantages and disadvantages of Alum and NanoAlum in these properties and their performance as adjuvants. Moreover, we propose the key features for ideal adjuvants and demonstrate that LDH NanoAlum is a promising candidate by summarizing its current progress in immunotherapeutic cancer treatments. Finally, we conclude the review by offering our integrated perspectives about the remaining challenges and future directions for NanoAlum's application in preclinical/clinical settings.

The Ly-6 and uPAR (LU) domain-containing proteins represent a large family of cell-surface markers. In particular, mouse Ly-6A/Sca-1 is a widely used marker for various stem cells; however, its human ortholog is missing. In this study, based on a systematic survey and comparative genomic study of mouse and human LU domain-containing proteins, we identified a previously unannotated human gene encoding the candidate ortholog of mouse Ly-6A/Sca-1. This gene, hereby named LY6A, reversely overlaps with a lncRNA gene in the majority of exonic sequences. We found that LY6A is aberrantly expressed in pituitary tumors, but not in normal pituitary tissues, and may contribute to tumorigenesis. Similar to mouse Ly-6A/Sca-1, human LY6A is also upregulated by interferon, suggesting a conserved transcriptional regulatory mechanism between humans and mice. We cloned the full-length LY6A cDNA, whose encoded protein sequence, domain architecture, and exon-intron structures are all well conserved with mouse Ly-6A/Sca-1. Ectopic expression of the LY6A protein in cells demonstrates that it acts the same as mouse Ly-6A/Sca-1 in their processing and glycosylphosphatidylinositol anchoring to the cell membrane. Collectively, these studies unveil a novel human gene encoding a candidate biomarker and provide an interesting model gene for studying gene regulatory and evolutionary mechanisms.
Humans ; Membrane Proteins/genetics* ; Pituitary Neoplasms/genetics* ; Biomarkers

Objective:To explore the application of venetoclax in transplantation of patients with refractory acute myeloid leukemia (AML).Methods:The diagnosis and treatment process of a patient with refractory AML who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) under venetoclax and hypomethylating agents bridging myeloablative preconditioning regimen after induction therapy failure in the First Affiliated Hospital of Soochow University in March 2020 were retrospectively analyzed.Results:The patient was a 28-year-old female who was diagnosed with refractory AML. The patient was initially given induction chemotherapy with IA (idarubicin+cytarabine) (3+7) regimen, but the disease did not relieve, then the induction chemotherapy with CLAG (cladribine+cytarabine+granulocyte colony stimulating factor) regimen was given, but the disease still did not relieve. After chemotherapy with venetoclax and hypomethylating agents bridging myeloablative preconditioning regimen, salvage haploid allo-HSCT was performed. Re-examination of bone marrow showed remission, and implantation was successful. The patient was followed up for 100 days and had sustained remission, and no transplantation complications occurred.Conclusion:For refractory AML patients who have failed primary induction therapy, the use of venetoclax and hypomethylating agents bridging myeloablative preconditioning regimen can be used as a preferred solution for salvage allo-HSCT.

Object To examine the preliminary clinical efficacy of custom-made three-dimensional(3D) printed talus prosthesis in the treatment of collapse talus necrosis. Methods:The clinical data of 8 patients who received 3D printed custom-made talus prostheses replacement for severe collapsed necrosis of the talus at the Orthopaedic Sports Medical Center, the First Affiliated Hospital to Army Medical University were analyzed retrospectively.All patients were male,with an average age of 38.0 years (range:22 to 65 years).There were 5 cases of left talus collapse and 3 cases of right talus collapse,with the course of disease of 29.7 weeks (range:6 to 96 weeks).The CT data of contralateral healthy talus were used for mirror image design references for the prosthesis,and the electron-beam 3D printing technology was used to prepare the prosthesis.Titanium alloy (Ti6Al4V) was taken as the material for the preparation of the talus body prosthesis,and Co-Cr-Mo material was used as the material for the preparation of the tibialis talus lateral joint surface prosthesis,and the subtalar joint surface of the prosthesis was made from a microporous casting technique.The prosthesis was analyzed preoperatively by digital three-dimensional finite element analysis and solid comparison techniques to measure anatomic match of the prosthesis.A longitudinal incision on medial ankle was made.The necrotic talus was completely removed and the prosthesis was then implanted.The patient was reexamined in the outpatient department 3, 6, and 12 months after surgery.Primary outcome measures were the American Orthopaedic Foot and Ankle Society(AOFAS) ankle-hind foot score,visual analogue scale(VAS) and ankle range of motion.Changes in imaging data and plantar pressure were also assessed.Repeated measures analysis of variance and paired- t test were used to compare the data. Results:The talus prosthesis measure preoperatively was completely consistent with that contralateral healthy talus and there was no operation-related complication. All the wounds healed primarily. The patients were followed up effectively for 23.17 months (range:12 to 48 months).The preoperative dorsiflexion of patients was (7.6±5.7)°,it increased to(14.2±6.6)° at 12 month after surgery ( t=-2.67, P=0.03).The plantar flexion increased from (22.0±9.9)°preoperatively to (29.2±8.7)° at 12 month after surgery ( t=-8.95, P<0.01).Preoperative AOFAS ankle-hind foot score was 26.3±6.6,and it increased to 70.1±2.2,76.0±3.4 and 79.3±4.2 at 3 month,6 month and 12 month after surgery( F=56.81, P<0.01);Pre-operative VAS was[ M( Q R)]3.0(0.8),and it increased to 2.5(1.0),1.5(1.0),1.0(1.0)at 3 month,6 month and 12 month after surgery( F=20.00, P<0.01).At the last follow-up,imaging reexamination showed that the prosthesis of all patients were in stable position with no sign of subsidence.No secondary ankle fusion or revision was required.The talus height increased from (27.6±6.0)mm preoperatively to (34.6±3.5)mm ( t=-2.94, P<0.01).The plantar pressure showed that the maximum pressure on the healthy ankle was(629.9±26.1)N,and that on the affected side was(521.4±14.4)N.The pressure on the healthy ankle was(350.6±29.6)N,and that on the necrotic side was (212.3±9.7)N.The load on the contralateral forefoot was(38.1±2.8)% and that on the necrotic side was(11.5±2.0)%.The load on the contralateral hindfoot was (24.6±2.5)% and that on the necrotic side was (21.1±1.8)%. Conclusions:The custom-made 3D printed talus prosthesis could restore the talus anatomy,recover the ankle joint function,relieve the pain of patients and improve the life quality of patients.The effect on plantar pressure is mainly achieved by adjusting the center of gravity of plantar pressure backwards and the increase of weight bearing of the healthy foot.

Object To examine the preliminary clinical efficacy of custom-made three-dimensional(3D) printed talus prosthesis in the treatment of collapse talus necrosis. Methods:The clinical data of 8 patients who received 3D printed custom-made talus prostheses replacement for severe collapsed necrosis of the talus at the Orthopaedic Sports Medical Center, the First Affiliated Hospital to Army Medical University were analyzed retrospectively.All patients were male,with an average age of 38.0 years (range:22 to 65 years).There were 5 cases of left talus collapse and 3 cases of right talus collapse,with the course of disease of 29.7 weeks (range:6 to 96 weeks).The CT data of contralateral healthy talus were used for mirror image design references for the prosthesis,and the electron-beam 3D printing technology was used to prepare the prosthesis.Titanium alloy (Ti6Al4V) was taken as the material for the preparation of the talus body prosthesis,and Co-Cr-Mo material was used as the material for the preparation of the tibialis talus lateral joint surface prosthesis,and the subtalar joint surface of the prosthesis was made from a microporous casting technique.The prosthesis was analyzed preoperatively by digital three-dimensional finite element analysis and solid comparison techniques to measure anatomic match of the prosthesis.A longitudinal incision on medial ankle was made.The necrotic talus was completely removed and the prosthesis was then implanted.The patient was reexamined in the outpatient department 3, 6, and 12 months after surgery.Primary outcome measures were the American Orthopaedic Foot and Ankle Society(AOFAS) ankle-hind foot score,visual analogue scale(VAS) and ankle range of motion.Changes in imaging data and plantar pressure were also assessed.Repeated measures analysis of variance and paired- t test were used to compare the data. Results:The talus prosthesis measure preoperatively was completely consistent with that contralateral healthy talus and there was no operation-related complication. All the wounds healed primarily. The patients were followed up effectively for 23.17 months (range:12 to 48 months).The preoperative dorsiflexion of patients was (7.6±5.7)°,it increased to(14.2±6.6)° at 12 month after surgery ( t=-2.67, P=0.03).The plantar flexion increased from (22.0±9.9)°preoperatively to (29.2±8.7)° at 12 month after surgery ( t=-8.95, P<0.01).Preoperative AOFAS ankle-hind foot score was 26.3±6.6,and it increased to 70.1±2.2,76.0±3.4 and 79.3±4.2 at 3 month,6 month and 12 month after surgery( F=56.81, P<0.01);Pre-operative VAS was[ M( Q R)]3.0(0.8),and it increased to 2.5(1.0),1.5(1.0),1.0(1.0)at 3 month,6 month and 12 month after surgery( F=20.00, P<0.01).At the last follow-up,imaging reexamination showed that the prosthesis of all patients were in stable position with no sign of subsidence.No secondary ankle fusion or revision was required.The talus height increased from (27.6±6.0)mm preoperatively to (34.6±3.5)mm ( t=-2.94, P<0.01).The plantar pressure showed that the maximum pressure on the healthy ankle was(629.9±26.1)N,and that on the affected side was(521.4±14.4)N.The pressure on the healthy ankle was(350.6±29.6)N,and that on the necrotic side was (212.3±9.7)N.The load on the contralateral forefoot was(38.1±2.8)% and that on the necrotic side was(11.5±2.0)%.The load on the contralateral hindfoot was (24.6±2.5)% and that on the necrotic side was (21.1±1.8)%. Conclusions:The custom-made 3D printed talus prosthesis could restore the talus anatomy,recover the ankle joint function,relieve the pain of patients and improve the life quality of patients.The effect on plantar pressure is mainly achieved by adjusting the center of gravity of plantar pressure backwards and the increase of weight bearing of the healthy foot.

Objective:To investigate the incidence, risk factors and survival of cGVHD patients in combination of a haploidentical donor supported with an unrelated umbilical cord blood for hematopoietic stem cells transplantation (haplo-cord-HSCT).Methods:300 hematological malignancies individuals who received dual transplantation were enrolled in the study between January 2012 and July 2016 at the department of Hematology in the First Affiliated Hospital of Soochow University. The clinical diagnosis and scoring the severity of cGVHD syndromes according the National Institutes of Health (NIH) consensus conference in the 2014 update. Cox proportional hazards regression was used to identify risk factors associated with transplant outcomes.Results:During follow-up with a median time of 26.4 months (range 0.2-61.8) post transplantation, the 1-year, 3-year and 5-year cumulative incidence of cGVHD was 26.3 % (95 % confidence interval [CI], 23.5 %~29.1 %), 30.3 % (95 % CI, 27.3 %~33.3 % ) and 32.2 % (95 % CI, 28.7 %~35.7 %). For all 73 patients with cGVHD, first-line or second-line treatment were given. During the follow-up period, 53 patients survived, and 20 patients died. In multivariate analysis, the cGVHD overall survival (GOS) were associated with thrombocytopenia(<100×109/L)(HR=0.103, 95 % CI 3 %-36.1 %, P<0.001). Conclusions:Our data suggest that, the 5-year cumulative incidence of cGVHD was 32.2 % after haplo-cord-HSCT with hematological malignancies. Thrombocytopenia (<100×109/L)was independent risk factors for GOS.

Objective To investigate the monitoring significance of WT1 gene level in the prognosis of acute myeloid leukemia (AML) patients with normal karyotype after hematopoietic stem cell transplantation (HSCT). Methods The clinical data of 115 AML patients with normal karyotype who were treated with HSCT from July 2009 to March 2017 in the First Affiliated Hospital of Soochow University were retrospectively analyzed. The dynamic detection of bone marrow WT1 gene was carried out by using reverse transcription_polymerase chain reaction (RT_PCR). According to the relative expression level median of WT1 gene before transplantation, the whole patients were divided into the two groups (

Objective: To study the specific killing effect of CD4 membrane protein targeted chimeric antigen receptor modified T (CAR-T) cell. Methods: The second generation CD4 targeted chimeric antigen receptor containing 4-1BB costimulation domain was insert into lentiviral vector through recombinant DNA technology. Lentivirus was prepared and packaged by 293T cells with four plasmids. Beads activated T cells were transduced with lentivirus and the transduction efficiency was checked with Protein L and flow cytometry. T cell subsets and IFN-γ concentrations were detected with probe-tagged antibody and cytometric bead assay. Results: ①The transduction efficiency of activated T cells with prepared lentivirus were 50.0%-70.0%. A subset of CD8⁺ T cell acquired dim expression of CD4 membrane protein after activation. CD4⁺T cell and CD8⁺CD4^dim T cell were gradually killed by CD4 targeted CAR-T post lentivirus transduction. ②The kill efficacy of CD4 targeted CAR-T cell and control T cell toward KARPAS 299 T cell at an E∶T ratio of 8∶1 for 24 h was (96.9±2.1)% and (11.2±3.1)%, CAR-T cell has a higher killing efficacy than control T cell (t=7.137, P=0.028). The IFN-γ concentrations in culture supernatant of CAR-T cell with K562-CD4 cell, CAR-T cell with K562 cell and CAR-T cell alone were (15 648±2 168), (1 978±354) and (1 785±268) pg/ml, CAR-T cell cocultured with K562-CD4 cell produced more IFN-γ than the other two controls (P<0.01). Conclusions CD4 targeted CAR-T has an immunophenotype of CD8⁺CD4^-T cell. CD4 targeted CAR-T cell has killing efficacy toward normal CD4⁺T cell and CD4⁺T lymphoma cell. CD4 targeted CAR-T cell also has a killing efficacy toward CD4^dim target cell.