ObjectiveTo establish a highly specific and sensitive triplex TaqMan quantitative PCR method for the detection of mouse hepatitis virus (MHV), mouse parvovirus (MPV), and reovirus type 3 (Reo-3) in laboratory mice. MethodsRecombinant plasmid standards were constructed using conserved genomic regions of the three target viruses. Specific primers and probes were designed, and a triplex TaqMan quantitative PCR (qPCR) system was optimized through preliminary experiments. Sensitivity was evaluated using 10-fold serial dilutions (10¹-10⁷ copies/μL). Intra- and inter-assay repeatability were assessed via reproducibility experiments. ResultsA strong linear correlation (R ²>0.99) was observed between copy number and Ct value at template concentrations of 10²-10⁷ copies/μL. Intra-assay and inter-assay coefficients of variation (CV) were both below 5%. No cross-reactivity was observed with mouse cytomegalovirus (MCMV), vesicular stomatitis virus (VSV), Sendai virus (SeV), or pneumonia virus of mice (PVM). ConclusionThe triplex TaqMan quantitative PCR method established in this study has high sensitivity, good repeatability, and strong specificity. It enables the rapid, specific, and sensitive detection of MHV, MPV, and Reo-3, allows the simultaneous detection of multiple pathogens in a single tube, and can be applied to the detection of clinical samples from laboratory animals and to epidemiological investigations, thereby providing effective scientific and technological support for the prevention and control of viral infection and the interruption of transmission.

AIM:To investigate the current status of astigmatism and its associated factors among children and adolescents from kindergarten through senior high school in Yangpu District, Shanghai, and to provide scientific evidence for the prevention and control of refractive errors.METHODS:A cluster sampling method was employed. From September to November 2024, students from 14 randomly selected schools in Yangpu District, Shanghai, were enrolled as study subjects. Autorefraction and ocular biometry were performed under non-cycloplegic conditions, and questionnaire surveys were administered. Comparisons of astigmatism degree, detection rate, and axis were made across age groups, educational stages, and sexes. Multiple linear and logistic regression analyses were conducted to identify factors associated with astigmatism.RESULTS:A total of 7 233 students aged 4-18 y were enrolled including 3 731 males, 3 502 females, with the mean age 10.13±4.18 y, astigmatism -0.76±0.82 D, and the detection rate(≤-1.00 D)was 29.23%. Both the degree and detection rate of astigmatism were correlated with age and educational stage, peaking in senior high school, with the most pronounced changes observed during primary school. Besides age, astigmatic refractive power was influenced by spherical equivalent(β=0.105, P<0.001), mean keratometry(β=-0.118, P<0.001), sex(with boys as the reference, girls: β=0.054, P<0.001), frequent sustained near work for 30-40 min(with “almost never” as the reference, β=-0.030, P=0.038), preterm birth(β=-0.023, P=0.044), maternal myopia(β=-0.028, P=0.020), and homework completion time on rest days(β=-0.035, P=0.041). The detection rate of astigmatism was associated with maternal myopia(OR=1.130, P=0.039), male sex(OR=0.773, P<0.001), shorter sleep duration on school days(OR=0.909, P=0.028), shorter sleep duration on rest days(OR=0.937, P=0.046), preterm birth(OR=1.382, P=0.012), frequent excessive head lowering(OR=1.216, P=0.045), homework completion time on rest days(OR=1.043, P=0.046), higher spherical equivalent(OR=0.891, P<0.001), and higher mean keratometry(OR=1.188, P<0.001).CONCLUSION:The degree and detection rate of astigmatism among children and adolescents from kindergarten to senior high school in Yangpu District, Shanghai are correlated with educational stage, and primary school may represent a critical period for astigmatism prevention and control. Male sex, maternal myopia, preterm birth, high-intensity near work, and insufficient sleep and other factors may be associated with the onset and progression of astigmatism. It is recommended that education and health departments strengthen visual health screening during primary school, while promoting behavioral interventions such as scientific eye-use habits and adequate sleep across different educational stages to delay the onset and progression of astigmatism.

Abstract To clarify the service targets, service contents, and safeguard mechanisms of the basic public service program for student vision health management, the study grounded in the current status of child and adolescent myopia prevention and control in China and integrated with the practical experience and innovative practices of Wuhan National Demonstration Area for Student Vision Health Management, takes education as the main thread and schools as the base, consolidating resources from multiple stakeholders including education administrative departments, schools, families, and professional technical service institutions. Centering on four core components—vision health education, monitoring and record keeping, risk prevention and control, and dynamic management, the study constructs a multi tiered basic public service system for vision health management of primary and secondary school students. It further proposes the use of an intelligent management platform and a "three tier color coded early warning" mechanism (red, yellow, blue) to prevent and control student myopia at the source, providing schools with replicable and operable implementation pathways to enhance the level of student vision health management.

Objective To analyze the correlation between white blood cell(WBC)and blood cell indexes in megaloblastic anemia(MA)patients.Methods A total of 79 patients diagnosed as MA by bone marrow cell morphology at Adicon Medical Laboratory Center from June 2023 to June 2024 were included in MA group,and another 40 healthy physical examination subjects during the same period were included in health group.The levels of WBC,red blood cell(RBC),platelet(PLT),hemoglobin(Hb)and mean corpuscular hemoglobin(MCH),mean corpuscular volume(MCV)were measured and compared between two groups.Spearman correlation was used to analyze the correlation of each parameter.Receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of WBC in MA.Results The levels of WBC,PLT,Hb and RBC in MA group were significantly lower than those in healthy group(P＜0.05),MCH and MCV levels were significantly higher than those of healthy group(P＜0.05).WBC was positively correlated with PLT,Hb and RBC(P＜0.05),and were negatively correlated with MCH and MCV(P＜0.05).ROC curve analysis showed that the area under the curve of WBC diagnosis MA was 0.926(95％CI:0.876-0.975).The levels of PLT,Hb and RBC in severe and above group were significantly lower than those in mild to moderate group(P＜0.05).The degree of anemia in MA patients was negatively correlated with the levels of PLT,Hb and RBC(P＜0.05),and the more severe the anemia,the lower the levels of PLT,Hb and RBC.Conclusion The changes of WBC and RBC are related to the occurrence of MA,and WBC can be used as a reference index for the diagnosis of MA.

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 analyze the correlation between white blood cell(WBC)and blood cell indexes in megaloblastic anemia(MA)patients.Methods A total of 79 patients diagnosed as MA by bone marrow cell morphology at Adicon Medical Laboratory Center from June 2023 to June 2024 were included in MA group,and another 40 healthy physical examination subjects during the same period were included in health group.The levels of WBC,red blood cell(RBC),platelet(PLT),hemoglobin(Hb)and mean corpuscular hemoglobin(MCH),mean corpuscular volume(MCV)were measured and compared between two groups.Spearman correlation was used to analyze the correlation of each parameter.Receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of WBC in MA.Results The levels of WBC,PLT,Hb and RBC in MA group were significantly lower than those in healthy group(P＜0.05),MCH and MCV levels were significantly higher than those of healthy group(P＜0.05).WBC was positively correlated with PLT,Hb and RBC(P＜0.05),and were negatively correlated with MCH and MCV(P＜0.05).ROC curve analysis showed that the area under the curve of WBC diagnosis MA was 0.926(95％CI:0.876-0.975).The levels of PLT,Hb and RBC in severe and above group were significantly lower than those in mild to moderate group(P＜0.05).The degree of anemia in MA patients was negatively correlated with the levels of PLT,Hb and RBC(P＜0.05),and the more severe the anemia,the lower the levels of PLT,Hb and RBC.Conclusion The changes of WBC and RBC are related to the occurrence of MA,and WBC can be used as a reference index for the diagnosis of MA.

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 investigate the application effect of high-altitude field-based teaching in acute mountain sickness.Methods:The medical students of the classes 2018 and 2019 majoring in clinical medicine were selected as subjects, and they were divided into conventional teaching group and field-based teaching group, with 20 students in each group. The students in the conventional teaching group received classroom teaching alone, and those in the field-based teaching group received high-altitude field-based teaching after theoretical lectures. The two groups were compared in terms of the theoretical knowledge of acute mountain sickness, the quality score of internship, and rescue operation score of acute mountain sickness, and questionnaire feedback and post-class discussion were performed among trainees and teachers to evaluate the high-altitude field-based teaching model. SPSS 19.0 was used for statistical analysis.Results:Compared with the conventional teaching group, the field-based teaching group had significantly higher scores of the theoretical knowledge of acute mountain sickness (91.72±4.34 vs. 86.10±5.15, P<0.001), the quality score of internship (89.64±5.21 vs. 83.51±2.38, P<0.001), and the rescue operation of acute mountain sickness [94.05 (89.54, 94.87) vs. 87.01 (84.33, 90.82), P<0.001]. Conclusions:High-altitude field-based teaching can improve the teaching effect of acute mountain sickness and cultivate the interest and learning enthusiasm of students, and therefore, it holds promise for wide application.

Objective:Glucorticoid therapy has the potential to mitigate immunogical effect of naxitamab. Ketamine is an anesthetic medica-tion and cause weak or shallow breathing. This article is to analyze the effect of modified conditioning regimen with substitution re-mifentanil for ketamine and without glucorticoid therapy on adverse events associated with naxitamab. Methods:Clinical data with naxit-amab infusion under modified conditioning regimen in Sun Yat-sen University Cancer Center between June 2023 and June 2024 were re-trieved to analyze adverse events and risk factors. Results:Overall,seventeen patients underwent 201 infusions. The most frequent adverse events were as follows:neurological pain (all grades) 93.0％,hypertension 55.7％,hypotension 34.8％,respectively. Bronchospasm and hyp-oxia were seen in 3.0％ and 10.9％ infusions,respectively. Fever occurred less frequently in the second cycle of infusion. No patients suspen-ded infusion due to severe adverse event. Conclusions:The infusion of naxitamab is tolerable under the modified conditioning regimen and adverse event is less than expected and controllable.

Objective:Glucorticoid therapy has the potential to mitigate immunogical effect of naxitamab. Ketamine is an anesthetic medica-tion and cause weak or shallow breathing. This article is to analyze the effect of modified conditioning regimen with substitution re-mifentanil for ketamine and without glucorticoid therapy on adverse events associated with naxitamab. Methods:Clinical data with naxit-amab infusion under modified conditioning regimen in Sun Yat-sen University Cancer Center between June 2023 and June 2024 were re-trieved to analyze adverse events and risk factors. Results:Overall,seventeen patients underwent 201 infusions. The most frequent adverse events were as follows:neurological pain (all grades) 93.0％,hypertension 55.7％,hypotension 34.8％,respectively. Bronchospasm and hyp-oxia were seen in 3.0％ and 10.9％ infusions,respectively. Fever occurred less frequently in the second cycle of infusion. No patients suspen-ded infusion due to severe adverse event. Conclusions:The infusion of naxitamab is tolerable under the modified conditioning regimen and adverse event is less than expected and controllable.