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

Excavating the quantitative trait locus (QTL) associated with rice cooking quality, analyzing candidate genes, and improving cooking quality-associated traits of rice varieties by genetic breeding can effectively improve the taste of rice. In this study, we used the indica rice HZ, the japonica rice Nekken2 and 120 recombinant inbred lines (RILs) populations constructed from them as experimental materials to measure the gelatinization temperature (GT), gel consistency (GC) and amylose content (AC) of rice at the maturity stage. We combined the high-density genetic map for QTL mapping. A total of 26 QTLs associated with rice cooking quality (1 QTL associated with GT, 13 QTLs associated with GC, and 12 QTLs associated with AC) were detected, among which the highest likelihood of odd (LOD) value reached 30.24. The expression levels of candidate genes in the localization interval were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR), and it was found that the expression levels of six genes were significantly different from that in parents. It was speculated that the high expression of LOC_Os04g20270 and LOC_Os11g40100 may greatly increase the GC of rice, while the high expression of LOC_Os01g04920 and LOC_Os02g17500 and the low expression of LOC_Os03g02650 and LOC_Os05g25840 may reduce the AC. The results lay a molecular foundation for the cultivation of new high-quality rice varieties, and provide important genetic resources for revealing the molecular regulation mechanism of rice cooking quality.
Quantitative Trait Loci ; Oryza/genetics* ; Plant Breeding ; Cooking ; Genetic Association Studies

Objective: To analyze the causes of death in Keqiao District, Shaoxing City in 2020, so as to provide the evidence for formulating the disease control strategy. Methods: The dead cases' gender, age and causes of death in Keqiao District, 2020, were collected from Shaoxing Municipal Public Health Information System, and the mortality was estimated and standardized by the 2020 Chinese standard population. The gender- and age-specific mortality and distribution of causes of death were analyzed, and the years of potential life lost (YPLL), average years of potential life lost per death (AYLL) and YPLL rate (YPLLR) were calculated to evaluate the life lost. Results: The crude and standardized mortality rates of registered residents in Keqiao District were 636.97/105 and 494.72/105, respectively. The crude and standardized mortality rates were 712.67/105 and 512.23/105 in men and 564.75/105 and 473.25/105 in women, and a higher mortality rate was seen in men than in women (χ2=59.628, P<0.001). The mortality reduced and then increased with age, and peaked among residents at ages of 85 years and older (13 910.90/105). Chronic disease was the main cause of death, accounting for 85.60% of all causes of death and showing a mortality rate of 545.27/105. Cancer, cardiopathy, cerebrovascular disease, injury and poisoning, and respiratory diseases were the five leading causes of death, and lung cancer, gastric cancer, colorectal cancer, liver cancer and pancreatic cancer were the five leading causes of cancer-related mortality. The highest YPLL was caused by cancers (13 015.00 person-years), with a YPLLR of 1.88%, and the highest AYLL was caused by injury and poisoning (10.37 years). Conclusion Chronic non-communicable diseases, such as cancer and cardio-cerebrovascular diseases, were main causes of death among residents in Keqiao District in 2020. The management of chronic diseases requires to be reinforced to improve the quality of life.

Objective:To describe for the determination of contents of metabolites of benzene compounds in urine sample by high performance liquid chromatography.Methods:After acidification with hydrochloric acid, metabolites in urine were first extracted by acetonitrile and isopropanol (V∶V, 9∶1) with excessive sodium chloride, then gradient separated on a C18 column and then determined by DAD detector.Results:There were good linear relationship between peak areas and injection quality in range of 2.00-100 mg/L ( r>0.999). The detection limit and quantitative limit of this method were 4.15-70.7 μg/L and 13.8-235 μg/L respectively. The precision for the analysis of urine was1.78%-8.23% ( n =6). The average recovery of metabolites was 85.4%-105.5% at thee spiked levels in the range of 2.00-100 mg/L. Conclusion:The accuracy and reproducibility obtained make this method useful for the biological monitoring of occupational exposure to toluene, xylene, styrene and ethylbenzene.

Objective:To analyze the direct economic burden of occupational asthma patients and provide economic basis for the government to rationally allocate health resources.Methods:In September 2019, colleted the case data of 53 patients diagnosed with occupational asthma who were hospitalized in our hospital from December 2008 to December 2018, and analyze the impact of gender, age, diagnosis time, ducation level, allergen type to the length of stay, hospitalization cost, medical technology diagnosis and treatment costs, western medicine costs, average daily hospitalization costs and other indicators.Results:The average length of hospitalization for occupational asthma patients was (38.7±8.1) days, and the average hospitalization cost was 14743 yuan, of which medical technology diagnosis and treatment costs, western medicine costs, and comprehensive medical service costs accounted for the top three, 42.5% (331977/781369) , 32.0% (249942/781369) , 19.6% (153268/781369) respectively. Hospitalization days for occupational asthma patients has decreased significantly in 2014-2018 ( P<0.05) . There were no significant differences in hospitalization costs, medical technology diagnosis and treatment costs, western medicine costs, and average daily hospitalization costs for occupational asthma patients caused by different allergens (isocyanates, persulfates and phthalic anhydrides) ( P>0.05) . Hospitalization days, hospitalization costs, medical technology diagnosis and treatment costs, western medicine costs, and average daily hospitalization costs of patients with moderate occupational asthma were significantly higher than those of mild patients ( P<0.05) . Conclusion:Early detection of occupational asthma patients and early intervention can reduce the economic burden on patients and society.

Objective:To describe for the determination of contents of metabolites of benzene compounds in urine sample by high performance liquid chromatography.Methods:After acidification with hydrochloric acid, metabolites in urine were first extracted by acetonitrile and isopropanol (V∶V, 9∶1) with excessive sodium chloride, then gradient separated on a C18 column and then determined by DAD detector.Results:There were good linear relationship between peak areas and injection quality in range of 2.00-100 mg/L ( r>0.999). The detection limit and quantitative limit of this method were 4.15-70.7 μg/L and 13.8-235 μg/L respectively. The precision for the analysis of urine was1.78%-8.23% ( n =6). The average recovery of metabolites was 85.4%-105.5% at thee spiked levels in the range of 2.00-100 mg/L. Conclusion:The accuracy and reproducibility obtained make this method useful for the biological monitoring of occupational exposure to toluene, xylene, styrene and ethylbenzene.

Objective:To analyze the direct economic burden of occupational asthma patients and provide economic basis for the government to rationally allocate health resources.Methods:In September 2019, colleted the case data of 53 patients diagnosed with occupational asthma who were hospitalized in our hospital from December 2008 to December 2018, and analyze the impact of gender, age, diagnosis time, ducation level, allergen type to the length of stay, hospitalization cost, medical technology diagnosis and treatment costs, western medicine costs, average daily hospitalization costs and other indicators.Results:The average length of hospitalization for occupational asthma patients was (38.7±8.1) days, and the average hospitalization cost was 14743 yuan, of which medical technology diagnosis and treatment costs, western medicine costs, and comprehensive medical service costs accounted for the top three, 42.5% (331977/781369) , 32.0% (249942/781369) , 19.6% (153268/781369) respectively. Hospitalization days for occupational asthma patients has decreased significantly in 2014-2018 ( P<0.05) . There were no significant differences in hospitalization costs, medical technology diagnosis and treatment costs, western medicine costs, and average daily hospitalization costs for occupational asthma patients caused by different allergens (isocyanates, persulfates and phthalic anhydrides) ( P>0.05) . Hospitalization days, hospitalization costs, medical technology diagnosis and treatment costs, western medicine costs, and average daily hospitalization costs of patients with moderate occupational asthma were significantly higher than those of mild patients ( P<0.05) . Conclusion:Early detection of occupational asthma patients and early intervention can reduce the economic burden on patients and society.

OBJECTIVETo observe the effects of acupuncture at opposite acupoints on expression of hepatocyte growth factor (HGF) in rats with skeletal muscle contusion, and to explore the mechanism of acupuncture at opposite acupoints on skeletal muscle contusion.

METHODSFifty-four Sprague Dawley (SD) rats were randomly divided into a blank group (6 rats), a model group (24 rats) and an opposing needling group (24 rats). The model group and opposing needling group were further divided into 1-day subgroup, 3-day subgroup, 5-day subgroup and 7-day subgroup, 6 rats in each one. No intervention was given in the blank group, while the model of skeletal muscle contusion was established in the model group and opposing needling group by self-made contusion device. 24 hours after contusion, electroacupuncture (EA) was applied at "Zusanli" (ST 36) and the corresponding points ofpoints at health side for 15 min, once a day. The subgroups of opposing needling group were treated for 1 day, 3 days, 5 days and 7 days, respectively. No treatment was given in the model group. Samples were collected in the subgroups 1 day, 3 days, 5 days and 7 days after treatment. The morphological change of injured gastrocnemius muscle was observed by using microscope after HE staining. The positive cell rate of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry. The expression levels of HGF protein and PCNA protein were observed by Western blot.

RESULTS① The results of HE staining showed that, 1 day after contusion, the inflammatory cells of gastrocnemius muscle in the opposing needling group were less than those in the model group; 3 days and 5 days after contusion, myoblasts and myotubes in the opposing needling group were more than those in the model group; 7 days after contusion, the neonatal muscle cells in the opposing needling group were more than those in the model group. ② The results of immunohistochemistry showed that, 1 day, 3 days and 5 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly higher than that in the model group (all<0.001); 7 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly less than that in the model group (<0.001). ③ The results of Western blot showed that, 1 day, 3 days and 5 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly higher than that in the model group (all<0.05); 7 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly lower than that in the model group (all<0.05).

CONCLUSIONAcupuncture at opposite acupoints could regulate the expression of HGF and promote the activation, proliferation, migration and differentiation of muscle satellite cells in rats with skeletal muscle contusion, which could speed up the process of skeletal muscle injury repair.

Objective To observe the effects of massage on inflammation,oxidative stress and autophagy during the repair of acute contusion of skeletal muscles so as to explore its biological mechanisms.Methods Forty-two adult Sprague-Dawley rats were randomly divided into a control group (n =6),a model group (n =18),and a treatment group (n =18).Acute contusion of the gastrocnemius muscles of the rats in the model and treatment groups was inflicted using a home-made impactor.Beginning forty-eight hours later,15 minutes of massage was administered daily for two weeks.After one,7 and 14 days of the massage treatment,the injured gastrocnemius was resected from 6 rats of both the model and treatment groups.Morphological changes were observed using haematoxylin and eosin (HE) staining.The serum content of tumor necrosis factor alpha (TNF-α),interleukin1β (IL-1 β),C reactive protein (CRP) and prostaglandin E2 (PGE2) were detected using enzyme-linked immunosorbent assay (ELISA).The serum content of superoxide (SOD) and malondialdehyde (MDA) were detected using spectrophotometry.The expression of microtubule-associated protein 1 light chain 3 (LC3),Bcl-2 homeodomain protein Beclin1 and ubiquitin binding protein P62 were detected using Western blotting.Results The HE staining showed more significant collapse and swelling of cells in the model group than in the control group at each time point.New muscle cells were observed at days 7 and 14 in the model group.At each time point,significantly better recovery was observed in the treatment group compared to the model group,with more new muscle cells and better cell morphology.According to the ELISA results,a significant increase in serum pro-inflammatory factors occurred in the model group compared to the control group and compared to the treatment group after one day and 7 days of treatment.The average serum content of SOD and MDA in the model group was significantly higher than in the control group,while the average serum content of SOD in the treatment group was significantly higher than in the model group and that of MDA was significantly lower.Western blotting showed a significant decrease in LC3 (Ⅱ/Ⅰ) and Beclin1,as well as a significant increase in P62 in the model group at each time point compared with the treatment group and the controls.Conclusion Inflammation and oxidative stress increase significantly in a skeletal muscle after injury,but autophagy decreases significantly.Massage can effectively reduce the inflammatory response and oxidative stress and promote autophagy,which leads to quicker repair of skeletal muscles.

Objective To explore the effects and mechanism of electroacupuncture (EA) on denervation-induced atrophy in rats. Methods A total of 18 male Sprague-Dawley rats were divided into sham group (n=6), model group (n=6) and EA group (n=6). The latter two groups were clamped right sciatic nerve to establish atrophy model of skeletal muscle. On the second day after modeling, EA group accepted electroacupuncture on right Zusanli (ST36) and Huantiao (GB30) for two weeks. Their gastrocnemius muscles were obtained after intervention, and the wet weight ratio of the gastrocnemius muscles was calculated. The cross-sectional area (CSA) and diameter of muscle fibers were measured after HE staining. The protein expression of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), 70-KD ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6k (p-p70S6k) was tested with Western blotting. The gene expression of mTOR and p70S6K was detected with real-time quantitative polymerase chain reaction. Results Compared with the sham group, the wet weight ratio of the gastrocnemius muscle, CSA and diameter of the muscle fibers decreased in the model group and EA group (P<0.001), which were more in EA group than in the model group (P<0.01); the protein expression of mTOR, p-mTOR, p70S6K and p-p70S6K increased in the model group (P<0.01), and increased more in EA group (P<0.05); the gene expression of mTOR and p70S6K increased in the model group (P<0.05) , and increased more in EA group (P<0.05).Conclusion Electroacupuncture delays the atrophy of denervated skeletal muscles, which may relate to activation of mTOR/p70S6K signal pathway to impact synthesis of skeletal muscle proteins.