BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

Objective:To explore the effects of the continuing nursing model based on "Internet +" in premature infants and their families.Methods:Premature infants and their families admitted to the Department of Neonatology, Wenzhou Central Hospital, from July 2021 to October 2022 were selected by convenience sampling. They were randomly divided into control group and observation group, each consisting of 108 pairs. Infants in the control group received routine nursing intervention, while those in the observation group received "Internet +" based continuing nursing based on the care provided to the control group. Physical growth indicators (weight, length, head circumference), Neonatal Behavioral Neurological Assessment (NBNA) scores, family caregiving knowledge and ability scores were measured before intervention (at birth) and after intervention (at 6 months of age) in both groups. The occurrence of delayed physical development and unplanned rehospitalization of premature infants at 6 months of age was also recorded.Results:After intervention, the physical growth indicators, NBNA scores, and family caregiving knowledge and ability scores of both groups improved, with the observation group higher than the control group, and the differences were statistically significant (all P<0.05). At 6 months of age, the observation group had lower rates of delayed physical development and unplanned rehospitalization compared to the control group, with statistically significant differences (all P<0.05) . Conclusions:The continuing nursing model based on "Internet +" can effectively improve the caregiving knowledge and ability of premature infants' families, promote the growth and development of premature infants, and reduce the risk of delayed physical development and rehospitalization.

Objective:To analyze the clinical manifestations and laboratory features in patients with myeloid neoplasms complicated with clonal T large granular lymphocyte (T-LGL) proliferation.Methods:The clinical data of 5 patients with myeloid neoplasms complicated with clonal T-LGL proliferation from November 2017 to November 2018 in Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College were analyzed retrospectively.Results:The median age was 60 years old. All patients had a history of abnormal peripheral blood cell counts for over 6 months. The absolute lymphocyte count in peripheral blood was less than 1.0×10 9/L. In addition to the typical T-LGL phenotype, the immunophenotype was heterogenous including CD4 +CD8 - in 2 patients, the other 3 CD4 -CD8 +. Four patients were αβ type T cells, the other one was γδ type. STAT3 mutation was detected in 1 patient by next-generation sequencing, the other 4 cases were negative. Conclusions:Clonal T-LGL proliferation with myeloid neoplasm develops in an indolent manner, mainly in elderly patients. Hemocytopenia is the most common manifestation. The diagnosis of T-LGL proliferation does not have specific criteria, that it should be differentiated from other T cell proliferative disorders, such as T-cell clones of undetermined significance. STAT3 or STAT5b mutation may help distinguish.

Objective:To explore the clinical characteristics of pediatric glucose transporter type 1 deficiency syndrome (GLUT1 DS), evaluate the efficacy and safety of ketogenic diet therapy (KDT).Methods:Clinical data of 19 children with GLUT1 DS admitted to Children′s Hospital of Fudan University, Tianjin Children′s Hospital, Shenzhen Children′s Hospital, Children′s Hospital of Nanjing Medical University and Jiangxi Provincial Children′s Hospital between 2015 and 2019 were collected retrospectively. The first onset symptom, main clinical manifestations, cerebrospinal fluid features and genetic testing results of patients were summarized, the efficacy and safety of ketogenic diet treatment were analyzed. Results:Among the 19 cases, 13 were males and 6 females. The age of onset was 11.0 (1.5-45.0) months,the age of diagnosis was 54.0 (2.8-132.0) months. Epilepsy was the first onset symptom of 13 cases. Different forms of tonic-clonic seizures were the most common types of epilepsy (7 cases with generalized tonic-clonic seizures, 5 cases with focal tonic or clonic seizures, 4 cases with generalized tonic seizures). Antiepileptic drugs were effective in 4 cases. Paroxysmal motor dysfunction was present in 12 cases and ataxia was the most common one. All patients had different degrees of psychomotor retardation. Among 17 patients received cerebrospinal fluid examination, cerebrospinal fluid (CSF) glucose level was lower than 2.2 mmol/L and CSF glucose/glycemic index was<0.45 in 16 cases, only 1 case presented normal CSF glucose level (2.3 mmol/L) and normal CSF glucose/glycemic index(0.47). SLC2A1 gene mutations were found in 16 patients, missense, frameshift and nonsense mutations were the common types with 5 cases, 5 cases and 3 cases respectively. All 19 patients were treated with ketogenic diet, which was effective in 18 cases in seizure control, 11 cases in dyskinesia improvement and 18 cases in cognitive function improvement. No serious side effects were reported in any stage of KDT.Conclusions:The diagnosis of GLUT1 DS is often late. It is necessary to improve the early recognition of the disease and perform CSF glucose detection and genetic testing as early as possible. The KDT is an effective and safe treatment for GLUT1 DS, but a small number of patients have not response to diet therapy.

Objective To investigate the effect of procyanidins on cerebral edema and inflammation of secondary brain injury after intracerebral hemorrhage.Methods Forty-eight C57BL6 mice were randomly divided into 4 groups:sham-operation+ saline group,sham-operation + procyanidins (100 mg/kg) group,intracerebral hemorrhage (ICH) + saline group and ICH + procyanidins (100 mg/kg) group.The Bederson method was used to detect postoperative nerve function score in mice.The brain tissue water content was measured by wet-dry weight method.The variety of morphological in brain tissue was observed by hematoxylin-eosin (HE) staining.The expression of glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1(Iba-1) in mouse brain tissue were observed by western blot.The apoptosis of brain tissue was observed by TUNEL and western blot method.Results Compared with the ICH+saline group after 12h,the nerve function score in the ICH + proanthocyanidins group was significantly decreased (P＜0.05),and the water content of brain tissue was also significantly decreased (P＜0.05).The HE staining results showed that procyanidins decreased the number of neurocytes lesions in cerebral hemorrhage mice and reduced the amount of brain tissue hemorrhage.Western blot results showed that procyanidins promoted the expression of Bcl-2,decreased the expression of Bax,Caspase3,GFAP and Iba-1 in cerebral hemorrhage mice.The TUNEL results showed that procyanidins could inhibit the apoptosis of brain tissue neurons in mouse with intracerebral hemorrhage.Conclusion Procyanidins inhibits the production of neurotoxic substances possibly by reducing the expression of GFAP and Iba-1,and prevents and treats secondary brain injury after intracerebral hemorrhage.

Objective To investigate the trans‐regulative effect of hepatitis B virus (HBV) preS2 on the promoter of human acyl protein thioesterase 1 (APT1) gene .Methods The promoter sequence of human APT1 gene was identified applying the soft‐ware of bioinformatics .The APT1 promoter and HBV preS2 gene were amplified with PCR and cloned into pGL3 and pcDNA3 .1 (-) plasmids to construct the luciferase reporter gene plasmid of human APT1 gene promoter pGL3‐APT1 and the preS2 eukary‐otic expression plasmid pcDNA3 .1(-)‐preS2 ,respectively .The effect of the preS2 on the human APT1 gene promoter was exam‐ined by cotransfecting hepatocellular carcinoma cell HepG2 with pGL3‐APT1 and pcDNA3 .1(-)‐preS2 and measuring luciferase activities of the HepG2 cells .The statistical data were analyzed with independent‐samples t test .Results Both plasmids of pGL3‐APT1 and pcDNA3 .1(-)‐preS2 were confirmed by DNA sequencing to be accurately constructed as design .The luciferase activity of the pGL3‐APT1 was 1 .2 times (P<0 .01) that of the positive control plasmid pGL3‐Control .And the luciferase activity of the HepG2 cells cotransfected with pcDNA3 .1(-)‐preS2 and pGL3‐APT1 was 2 .6 times (P<0 .01) that of the HepG2 cells cotrans‐fected with the plasmid without preS2 gene pcDNA3 .1(-) and pGL3‐APT1 .Conclusion The human APT1 promoter cloned in the study has high promoter activity ;HBV preS2 activates human APT1 promoter .

Objective:To evaluate the value of transesophageal echocardiography(TEE)in postoperative low cardiac output syndrome of critical cardiac valvular disease.Methods:The preoperative data of 16 patients with critical cardiac valvular disease who suffered from postoperative low cardiac output syndrome,as well as the postoperative treatment of them,was retrospec-tively analyzed.Results:Among the 16 patients,9 cases underwent intra-aortic balloon pump(IABP)support suggested by TEE.And 7 of the 9 patients recovered without complications,while the other 2 patients died.IABP were not suggested by TEE in 7 patients,2 of whom underwent exploratory thoractomy,while the other 5 patients recovered after adjustment of vas-opressor agents and improvement of internal environment.Conclusions:TEE can be used in the differentiation of the cause of postoperative low cardiac output syndrome in patients with critical cardiac valvular disease and it can provides the basis for the reasonable application of IABP.

ObjectiveTo observe the effects of different angles between tibial tunnel and tibial platform on “killer turn” in posterior cruciate ligament (PCL) reconstruction,and primarily discuss a safe and reasonable tunnel technology. Methods Eighteen fresh tendon grafts were used to reconstruct the PCL on the tibial side of fresh cadavers.The tibial tunnels of all specimens were built via anteromedial approach.Based on the different angles between tibial tunnel and tibial platform,all specimens were divided into Group A (30°),Group B (40°) and Group C (50°),with six specimens in each group.Area of tibial tunnel exit,pressure of tibia tunnel exit and circulation characteristics of tendons under the cyclic load before and after biomechanical test were recorded.ResultsThe area of tibial tunnel exit had statistical difference among three groups after the test ( F =8.80,P ＜ 0.05 ).The pressure of tibial tunnel exit had statistical difference among three groups (F =3.91,P ＜ 0.05 ).The cyclic frequency and fatigue strength of the transplanted tendons had statistical difference among three groups under the same cyclic load ( 256 N ) and same frequency ( 126 Hz ) ( F =4.25,P ＜ 0.05 ).Conclusions The angle between tibial tunnel and tibial platform has negative correlation with the area and pressure of tibial tunnel exit,and has positive correlation with the cyclic frequency and fatigue strength of the transplanted tendons under cyclic load.The ideal anatomy position of the tibial tunnel is the anteromedial tunnel with the angle of 40° between the tibial tunnel and the tibial platform.

Objective To screen and identify the differential expression genes on peripheral blood cells of mice based on the experimental animal model of radon exposure. Methods BALB/c mice were exposed in a type HD-3 multifunctional radon-room, with the accumulative doses of radon-exposure group at 105 WLM and control group at 1 WLM. Total RNA was extracted from peripheral blood cells and the methods of SMART for dscDNA synthesis and SSH for gene screening was applied. With the construction of the cDNA library enriched with differentially expressed genes, the pMD 18-T plasmid containing LacZ operator at the multiple cloning site was used to allow a blue-white screening. The TA clones were amplified by nested PCR and the reverse Northern blot was used to identify up and down regulation of the clones. The differently expressed cDNA was then sequenced and analyzed. Results The subtracted cDNA libraries were successfully constructed. A total of 390 recombinant white colonies were randomly selected. Among the 312 cDNA monoelones selected from bath forward- and reverse-subtracted libraries,41 clones were chosen to sequence for their differential expressions based on reverse Northern blot. Among the 41 sequenced clones, 10 clones with known function/annotation and 3 new ESTs with the GenBank accession numbers were obtained. Most of the known function/annotation genes were revealed to be related with cell proliferation, metabolism, cellular apoptosis and carcinogenesis. Conclusions The animal model of radon exposure was established and the cDNA library of peripheral blood cells was suceessfully constructed. Radon exposure could up- and down-regulate a series of genes. Differentially expressed genes could be identified by using SSH technique and the results may help exploring mechanisms of random exposure.

Objective: To investigate the relationship between C-reactive protein(CRP)and its-717MG polymorphismin atrial fibrillation(AD of the population of the city of Nantong.Methods:The relationships between AF and AF risk factors were analyzed by comparing genders,ages and body nlagS index(BWI)in 92 AF patients and 60 non-AF control subjects.The serum CRP levels were detected by immunoturbidimetry in of the two groups.The CRP-717MG polymorphism wa8 detected by polymerage chain reaction-restriction fragment-length polymorphism in patients and control subjects.Results:The sel3utm CRP level wag positively correlated with the left atrium internal diameter(LAD)in AF patient group(r=0.58,P＜0.01).The level of CRP Wag significantly higher in AF patient group compared with that of control group(P＜0.01).The serum CRP level Wag higher in patients with non-paroxysmal atrial fibrillation than that of patients with paroxysmal atrial fibrillation(P＜0.05).There Wag no significant difference in the frequency of CRP genotype between AF and control groups(P＞0.05).But the alleles frequency of the G Wag lower in AF group than that of control group(P＜0.05).Conclusion:The semm CRP level is associated with AF and its subgroups.The serum CRP level is positively correlated with LAD.The results suggest that the inflammation influences the AF though atrium reconstruction.The relationship between CRP-7 17A/G and AF stir needs further large-scale perspective studies.