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

Phage immunoprecipitation sequencing (PhIP-Seq) is a high-throughput and low-cost method for analyzing the specific binding of target proteins to peptide libraries. The method uses oligonucleotide library synthesis (OLS) to encode proteome-scale peptide libraries for display on phages, and then immunoprecipitates these library phages with target proteins (such as antibodies) for subsequent analysis by high-throughput DNA sequencing. PhIP-Seq enables the screening of peptide targets that react specifically with hundreds of proteins or pathogens. PhIP-Seq has been successfully applied in various fields such as disease detection, screening of autoimmune disease biomarkers, vaccine development, and allergen detection, becoming a high-throughput diagnostic technology. This article systematically describes the development, applications, and result evaluation of PhIP-Seq, in order to gain a more comprehensive understanding of the application and future development prospects of this technology in various fields.
Peptide Library ; Humans ; Immunoprecipitation/methods* ; High-Throughput Nucleotide Sequencing/methods* ; Bacteriophages/genetics*

ObjectiveBy starting with the combination of Os Draconis， Bupleuri Radix， and Ostreae Concha， the role of mineral medicine Os Draconis in the combination of the Bupleuri Radix-containing tri-herbal medicines was preliminarily explored from the perspective of supramolecular system formation. Method① The appearance and Tyndall phenomenon of single decoction of Os Draconis， Bupleuri Radix， and Ostreae Concha， as well as co-decoction of Bupleuri Radix-Os Draconis， Bupleuri Radix-Os Draconis-Ostreae Concha， and Bupleuri Radix-Ostreae Concha were observed， and the average particle size， dispersion coefficient， and Zeta potential of suspension particles in each decoction were determined. The micromorphology of supramolecular structures was observed by scanning electron microscope （SEM）. ② The pH of different compatibility systems， liquid viscosity coefficient， liquid surface tension， freeze-dried powder yield rate， and other physical properties were determined， and the interaction of different compatibility systems was detected by infrared absorption spectroscopy （FTIR） and UV-visible spectrophotometry （UV-Vis）. ③ The composition and content difference of different compatible systems were determined by high-performance liquid chromatography （HPLC） and ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry （UPLC-Q-TOF-MS）. ResultCompared with the single decoction， the co-decoction had more obvious turbidity and Tyndall phenomenon. The particles in the co-decoction suspension were smaller and more evenly distributed， and the Zeta potential was reduced， indicating a more stable system. Under SEM， Bupleuri Radix was irregularly lamellar， and Bupleuri Radix-Os Draconis and Bupleuri Radix-Os Draconis-Ostreae Concha were mainly spherical nanoparticles. Bupleuri Radix-Ostreae Concha was irregularly lamellar， with a small number of spherical nanoparticles. The pH of the single decoction of Bupleuri Radix and co-decoction increased， and the viscosity coefficient increased. The liquid surface tension decreased. The freeze-dried powder yield rate of the Bupleuri Radix-Os Draconis co-decoction was the highest， followed by Bupleuri Radix-Ostreae Concha decoction and Bupleuri Radix-Os Draconis-Ostreae Concha decoction， and the yield rate of Bupleuri Radix single decoction was the lowest. The main change of FTIR was the stretching vibration of -OH， and the co-decoction moved to the low-frequency direction obviously. UV-Vis showed that the maximum absorption occurred at 295.8 nm for all groups， and the absorption intensity was different （Bupleuri Radix-Os Draconis>Bupleuri Radix-Os Draconis-Ostreae Concha>Bupleuri Radix-Ostreae Concha>Bupleuri Radix）. The components of Bupleuri Radix were used as the indexes， and the content of methanol extract determined by HPLC was higher than that of water extract， and the components of Bupleuri Radix single decoction were mainly saikosaponin a （SSa） and saikosaponin c （SSc）， which were slightly higher after co-decoction compatibility. UPLC-Q-TOF-MS could identify 37 compounds in both single decoction and co-decoction. ConclusionThe combination of Bupleuri Radix， Os Draconis， and Ostreae Concha can form a smaller， more uniform， and stable nano-sized supramolecular system， which is conducive to the dissolution of the main components of Bupleuri Radix， and the Os Draconis contributes the most in this process.

Objective:To design a systematic emotion management training (EMT) targeted the characteristics of coping strategies based on rehabilitation assistant and explore the feasibility of online EMT in improving cognitive coping strategies, mental health, and social support of patients with coronary heart disease(CHD).Methods:From June 2021 to December 2022, a total of 106 CHD patients were screened from cardiovascular units of a hospital in Zhengzhou. All participants were divided into the intervention group ( n=53) and control group ( n=53) using the coin toss method. The patients in intervention group received 7 weeks emotion management training on the basis of routine health education, the patients in control group received 7 weeks routine health education. The scores of cognitive coping strategies, anxiety, depression, and social support were measured at baseline and post-intervention. Statistical analysis was conducted using SPSS 24.0 software.The cognitive coping strategies, mental health and social support scores between two groups were compared using independent sample t-tests or paired t-test. Pearson analysis was used to examine the correlation between cognitive coping strategies, mental health and social support. Results:The scores of adaptive coping strategies, maladaptive coping strategies, depression, anxiety, insomnia and social support in patients with CHD were (69.52±7.60), (35.22±6.15), (8.82±2.66), (8.78±1.99), (10.97±2.86), and (57.48±9.79), respectively. After intervention, the scores of maladaptive coping strategies, self-blame, rumination, catastrophizing, blaming others, putting into perspective, insomnia, anxiety, and depression in the intervention group were significantly lower than those in the control group( t=-7.742, -4.395, -4.781, -6.105, -6.256, -5.327, -6.017, -7.288, -7.749, all P<0.05). The scores of adaptive coping strategies, positive reappraisal, refocusing on planning and social support were significantly higher than those of the control group( t=7.314, 6.733, 5.874, 3.562, all P<0.05). In the intervention group, there were statistically significant differences in cognitive coping strategies, anxiety, depression, insomnia and social support scores before and after the test(all P<0.05) and they were not statistically significantly different in the control group(all P>0.05). Correlation analysis showed that maladaptive coping strategies were positively correlated with depression and anxiety scores ( r=0.421, 0.408, both P<0.05). Adaptive coping strategies were negatively correlated with depression and insomnia scores ( r=-0.225, -0.240, both P<0.05), and positively correlated with social support ( r=0.219, P=0.034) among patients with CHD. Conclusion:The online EMT based on rehabilitation assistant may be a useful intervention for patients with CHD which can promote psychological health, social support as well as positively transforming maladaptive coping strategies into adaptive coping strategies.

AIM: To analyze the causes of blindness and low vision in patients with visual disability in Yangpu District of Shanghai from 2019 to 2022.METHODS:Cross-sectional study. A total of 1 604 patients who participated in the evaluation of visual disability in Shanghai Yangpu District Kongjiang Hospital, from April 2019 to December 2022 were selected for the study. The grade of visual disability and the main causes of blindness and low vision were determined by trained doctors.RESULTS:A total of 804 patients with visual disabilities were identified, with 87.31% aged 60 and above. The causes of visual disability were high myopic retinopathy(30.47%), age-related macular degeneration(23.26%), glaucoma(17.04%), and diabetic retinopathy(11.07%). Glaucoma(36.96%)is the leading cause of blindness.CONCLUSION: The majority of patients with visual disability are aged 60 years and above. More attention should be paid to the elderly population. Comprehensive prevention, treatment and rehabilitation measures should be applied in different diseases based on classification, so as to early reduce the occurrence of visual disability.

Objective:To study the trinucleotide repeats of GCN (GCA, GCT, GCC, GCG) encoding Alanine in exon 3 of the PHOX2B gene among healthy individuals from southwest China and two patients with Congenital central hypoventilation syndrome (CCHS). Methods:The number and sequence of the GCN repeats of the PHOX2B gene were analyzed by capillary electrophoresis, Sanger sequencing and cloning sequencing of 518 healthy individuals and two newborns with CCHS, respectively. Results:Among the 1036 alleles of the 518 healthy individuals, five alleles were identified, including (GCN) 7, (GCN) 13, (GCN) 14, (GCN) 15 and (GCN) 20. The frequency of the (GCN) 20 allele was the highest (94.79%). And five genotypes were identified, which included (GCN) 7/(GCN) 20, (GCN) 13/(GCN) 20, (GCN) 14/(GCN) 20, (GCN) 15/(GCN) 20, (GCN) 20/(GCN) 20. The homozygous genotypes were all (GCN) 20/(GCN) 20, and the carrier rate was 89.58%. Four GCN sequences of the (GCN) 20 homozygous genotypes were identified among the 464 healthy individuals. The GCN repeat numbers in the exon 3 of the PHOX2B gene showed no significant difference between the expected and observed values, and had fulfilled the, Hardy-Weinberg equilibrium. The genotypes of the two CCHS patients were (GCN) 20/(GCN) 25 and (GCN) 20/(GCN) 30, respectively. Conclusion:It is important to determine the GCN repeats and genotypic data of the exon 3 of the PHOX2B gene among the healthy individuals. The number of GCN repeats in 518 healthy individuals was all below 20. The selection of appropriate methods can accurately detect the polyalanine repeat mutations (PARMs) of the PHOX2B gene, which is conducive to the early diagnosis, intervention and treatment of CCHS.

Objective:To explore the changes and influencing factors of prosocial behavior in children with leukemia during different stages of treatment, using latent growth curve models and latent class growth models.Methods:A total of 120 children with leukemia and their parents were selected as subjects from 2022 to 2023. All the children were evaluated at four assessment time points (diagnosed within 1 week, at the end of 1 month, 3 months and 6 months after diagnosed, denoted as T1-T4) using self-designed survey scale, strengths and difficulties questionnaire-prosocial behavior(SDQ-PSB), family closeness scale, and posttraumatic growth inventory.SPSS 23.0 and Mplus 7.4 softwares were used to establish latent growth curve models and latent class growth curve models, and to analyze the trajectory of prosocial behavior and potential classes, as well as to explore their influencing factors.Results:The SDQ-PSB scores of the children at T1-T4 were (1.24±0.46), (2.79±0.22), (3.88±0.36), and (5.88±0.32), respectively.The total score of SDQ-PSB at T1-T4 showed significant positive correlation ( r=0.32-0.79, all P<0.01). Children with leukemia were divided into two latent classes based on their prosocial behavior: the persistent improvement group and the abnormal group.The initial SDQ-PSB scores of the abnormal group were lower than those of the persistent improvement group, and there was no significant change in SDQ-PSB scores over time (slope mean=0.2, P>0.05). In contrast, the SDQ-PSB scores of the persistent improvement group showed a significant increase over time (slope mean=0.8, P<0.001). Children under 7 years old were more likely to belong to the persistent improvement group ( B=-0.64, OR=0.62, 95% CI=0.15-0.71, P=0.003). Under certain conditions, an increase in family closeness ( B=-0.84, OR=0.69, 95% CI =0.49-0.72, P=0.003) and enhanced posttraumatic growth ( B=-0.52, OR=0.86, 95% CI=0.83-0.95, P=0.025) also led to development towards the persistent improvement group. Conclusion:The trajectory of prosocial behavior development in children with leukemia after diagnosis is abnormal and marginal, but it shows a significant quadratic upward trend with significant group heterogeneity as the disease progresses. Influencing factors include age, family income, family closeness, and levels of posttraumatic growth.

ObjectiveTo identify the members of the Rehmannia glutinosa miR166 gene family and clarify the response mode under adversity. MethodHigh-throughput sequencing technology was employed to obtain a small RNA database and the miR166 family members of R. glutinosa were screened out. The precursor structures were analyzed by RNAfold. DNAMAN and MEGA were used for conservative and evolutionary analyses， respectively. TargetFinder software was used to predict the target genes of R. glutinosa miR166 family members. The expression of miR166 family members in response to abiotic stress was analyzed by real-time polymerase chain reaction（Real-time PCR）. ResultFive miR166s were identified with precursors possessing complete stem-loop structures. As revealed by sequence alignment results， the precursors and matures were both highly conserved. Forty-eight target genes of miR166s were predicted， which were mainly annotated to the HD-ZIP Ⅲ family transcription factors. The expression characteristics showed that the expression of miR160s was up-regulated after R. glutinosa was infected by endophytic fungi， which was different from the expression of the family members under abiotic stress. The expression level of rgl-miR166b-5p in the drought-flood treatment group and the high-low temperature treatment group was significantly down-regulated compared with that in the control group， and the expression pattern was opposite under the endophytic fungal infection. ConclusionThe results of this study preliminarily clarified the expression patterns of R. glutinosa in response to biotic and abiotic stresses and provided a theoretical basis for future breeding and improvement of R. glutinosa.

To analyze the clinical features and genetic variants of a child with type 4 familial partial lipodystrophy (FPLD4) and the initial manifestation of diabetes.The male patient with the age of 13 years and 5 months, and the diabetes course was about 3 years, the patient was admitted to Children′s Hospital of Soochow University on November 10, 2021(4 th hospitalization at the hospital), in the course of diabetes, the children repeatedly suffered from diabetes ketoacidosis, and lipid metabolism complications gradually emerged.The gene sequencing showed that the proband and his mother carried dual gene mutations of PLIN1 c. 1325delG(p.G442Afs*99) and SPINK1 c. 194+ 2T>C(p.? ). The PLIN1 gene was the causal gene of FPLD4.The mutations of c. 1325delG in the PLIN1 gene had not been previously reported.Based on the clinical phenotype, family history and genetic testing findings, the patient was diagnosed as FPLD4.In addition, the mutation of SPINK1 c. 194+ 2T>C(p.? ) might increase the risk of chronic pancreatitis.This case report enriched the clinical characteristics and genotype data of FPLD4.Gene sequencing assisted the accurate diagnosis of the type of diabetes.The effects of dual gene mutations on disease progression should be concerned, which were of great significance to develop treatment regimen and disease management.

Objective:To evaluate the safety and feasibility of applying injectable cross-linked sodium hyaluronate isolation gel in radical hypofractionated radiation therapy for prostate cancer.Methods:In this prospective study, patients at Beijing Hospital who were pathologically diagnosed with clinical stage T 1-2N 0M 0 prostatic acinar adenocarcinoma by puncture and underwent radical radiation therapy were included. All patients received ultrasound-guided cross-linked sodium hyaluronate isolation gel injection and image-guided intensity-modulated radiation therapy (IG-IMRT). The prescription dose was moderately hypofractionated, with a prescription dose of 60 Gy in 20 fractions for 5 times a week, once daily, which was delivered to 95% of the planning target volume (PTV) of prostate and seminal vesicle. Analyze the prostate rectal spacing (PRS) at the baseline, on the day of injection, during the radiotherapy, 1 month and 3 months after radiotherapy, changes in rectal volume before and after injection, and incidence of rectum-related side effects. The changes in all indexes before and after injection were analyzed by using t-test. Results:A total of 13 patients were enrolled from March 2022 to February 2023. The isolation gel maintained morphologic stability without significant spatial changes during radiotherapy, and the mid-prostate had the best effect, with PRS up to 1 cm. At 3 months after radiotherapy, the isolation gel was seen to decreased in volume with a certain absorptive capacity. The irradiated volume of rectum was decreased significantly in all patients after gel injection, and the mean volumes of rectal V 60 Gy , V 50 Gy , V 30 Gy , and V 20 Gy before and after injection were 1.923% vs. 0.280%, 10.255% vs. 3.172%, 29.602% vs. 18.800%, and 49.452% vs. 40.259% (all P<0.005). The average values (range) of rectal V 60 Gy , V 50 Gy , V 30 Gy , V 20 Gy decreases were 84.9%( 29% - 100%), 69.6%(27%-100%), 36.3%(0%-75%), and 17.8%(0%-50%), respectively. No grade 3-4 side effects occurred in all patients, and there were no common grade 1-2 rectal side effects such as diarrhea, rectal bleeding, proctitis and anal pain, etc. Only one patient developed grade 1 constipation during radiotherapy. Conclusion:Injection of Chinese made cross-linked sodium hyaluronate isolation gel can significantly reduce the irradiated volume of rectum and the incidence of rectal toxicities in prostate cancer patients undergoing radical radiotherapy.