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 investigate the effect of Gouteng Jiangya Jieyu Perscription(Uncariae Ramulus cum Uncis,Gastrodiae Rhizoma,Pheretima,Puerariae Lobatae Radix,etc.)modulating the TLR4/NF-кB signaling pathway on the polarization of hippocampal microglia in rats with hypertension complicated with depression(HD)Methods Forty primary hypertensive rats were randomly divided into five groups:the model group,the positive drug group,and the high-,medium-,and low-dose groups of Gouteng Jiangya Jieyu Perscription,with 8 rats in each group;and another 8 SD rats were taken as the control group.The HD model was replicated using 42 days of continuous chronic unpredictable mild stress(CUMS)combined with solitary rearing.The modeling was accompanied by the administration of drugs,including 29.61,14.81,and 7.40 g·kg-1 of Gouteng Jiangya Jieyu Perscription in the high-,medium-,and low-dose groups of Chinese herbal medicine,respectively,and 0.45 mg·kg-1 of Levamlodipine Besylate+1.8 mg·kg-1 of Fluoxetine in the positive group;the volume of the gavage was 10 mL·kg-1,once a day,for 42 consecutive days.The systolic blood pressure of rat tail artery was measured by non-invasive sphygmomanometer before drug administration and in the morning of the last day of each week;the behavioural test of Sucrose Preference Test(SPT)was carried out once in the second week and once in the last week after the start of the modelling;the water maze experiment was carried out after the end of the modelling;the levels of serum inflammatory factor tumor necrosis factor-α(TNF-α),interleukin(IL)1β and IL-10 were determined by ELISA;the pathological changes of rat hippocampal tissue neurons were observed by HE staining and Nissl stain were used to observe the neuronal pathological changes in rat hippocampal tissue;immunofluorescence double staining was used to detect the expressions of microglia M1(CD16)and M2(CD206)types in the hippocampal region;and Western Blot was used to detect the protein expressions of TLR4 and NF-кB p65 in the hippocampal tissue.Results Compared with the control group,the systolic blood pressure in the tail artery of rats in the model group from week 1 to week 6 were all significantly increased(P＜0.01);sucrose preference rate was significantly decreased(P＜0.01);evasion latency was significantly prolonged(P＜0.05,P＜0.01),the number of times of traversing the plateau and the percentage of time spent in the target quadrant were significantly decreased(P＜0.01);the contents of serum TNF-α and IL-1β were significantly increased(P＜0.01),IL-10 content was significantly decreased(P＜0.01);cytosolic nuclei were deeply stained,cytoplasmic solidification and apoptosis were obvious;the fluorescence intensity ratio of CD206/CD16 in hippocampal microglial cells were significantly decreased(P＜0.05);the protein expressions of TLR4 and NF-кB p65 in hippocampal tissues were significantly up-regulated(P＜0.01).Compared with the model group,the systolic blood pressure in the tail artery of rats in the first to sixth weeks of the drug administration group were all significantly reduced(P＜0.01);the sucrose preference rates were all significantly increased(P＜0.05);the contents of serum TNF-α and IL-1β were significantly decreased(P＜0.01),and the content of IL-10 was significantly increased(P＜0.01);the Nissl substances are abundant and apoptosis is significantly reduced,and apoptosis were significantly reduced.The escape latency of rats in the positive drug group and the high-dose group of Gouteng Jiangya Jieyu Perscription was significantly shortened(P＜0.05,P＜0.01),and the number of times of crossing the plateau and the percentage of time spent in the target quadrant were significantly increased(P＜0.05);the ratio of fluorescence intensity of hippocampal microglial cells,CD206/CD16 was significantly increased(P＜0.05,P＜0.01);and the hippocampal tissues,protein expressions of TLR4 and NF-κB p65 were significantly down-regulated(P＜0.05,P＜0.01).Conclusion Gouteng Jiangya Jieyu Perscription may regulate the polarisation state of hippocampal microglial cells,modulate the secretion of inflammatory factors,and attenuate the damage of hippocampal neurons in HD rats by inhibiting the TLR4/NF-кB pathway.

Objective:To apply knowledge maps to systematically evaluate problem-based learning (PBL) literatures from the past 50 years, so as to explore the law of development and related important documents.Methods:All peer-reviewed studies were retrieved from Web of Science (WOS) Citation Database on PBL of the year from 1970 to 2019. Bibliometrix package of R was used to analyze following items: annual growth law of PBL literature, literature cumulative growth law, literature citation status and top 30 important documents. Based on the results, the law of development of PBL research, important literatures and time nodes was analyzed.Results:After literature cleaning, 2 401 documents were finally included into the analysis. The analysis revealed that: in accordance with the law of development disciplines, PBL study experienced three periods, early birth (Period Ⅰ), PBL study early development (Period Ⅱ) and PBL large-scale development (Period Ⅲ). Now PBL is currently in a period of large-scale development. In the development process, 1983, 1986, 1993, 2003, 2005, 2007, 2011, 2012 and 2014 has become important research time nodes, and there have been five important documents promoting PBL development (ALBANESE MA, 1993, ACAD MED; VERNON DTA, 1993, ACAD MED; NORMAN GR, 1992, ACAD MED; BARROWS HS, 1986, MED EDUC; HMELO-SILVER CE, 2004, EDUC PSYCHOL REV).Conclusion:Nowadays, PBL theory and method has been perfected, which has been developed significantly, and it's an important period of application and promotion. The PBL has become an important method to cultivate clinical skills or other disciplines with strong practicality.

Objective:To investigate the clinical value of shear wave elastography (SWE) in predicting pathological responses to neoadjuvant chemotherapy in breast cancer.Methods:According to the postoperative pathological responses, 56 patients who received neoadjuvant chemotherapy followed by surgical excision in the Fujian Cancer Hospital from August 2019 to September 2020 were divided into responders and non-responders. The relative change rates of tumor maximum diameter(ΔD2, ΔD4) and SWE stiffness (ΔEmax2, ΔEmax4, ΔEmean2, ΔEmean4) were assessed before NAC and after different NAC cycles (t2, t4). Clinical information, including age, T, N stages, ER, PR, HER2, Ki67, and molecular subtype were also considered as the variables. The independent influencing factors of pathological responses after neoadjuvant chemotherapy were obtained by logistic regression analysis and diagnostic test was carried out.Results:There were 23 cases as responders (41.0%, 23/56), and 33 cases as non-responders (58.9%, 33/56). Results of multivariate analysis showed ΔEmax4 and HER2 index were independent influencing factors of pathological responses ( OR=1.11, P<0.001; OR=31.81, P=0.002). Area under curve of the ΔEmax4 (AUC: 0.869, 95% CI: 0.746-0.941) was higher than that of HER2 (AUC: 0.690, 95% CI: 0.545-0.834). The combination of ΔEmax4 and HER2 gave the best prediction of pathological responses (AUC 0.930, 95% CI: 0.829-0.981). the sensitivity, specificity, diagnostic accuracy, postive predictive value, and negative predictive value were 78.26%, 96.97%, 75.23%, 94.73%, and 86.49%, respectively. Conclusions:ΔEmax4 and HER2 are independent predictors of pathological responses after neoadjuvant chemotherapy for breast cancer. Combined ΔEmax4 and HER2 can improve the predictive diagnostic efficacy of pathological responses to chemotherapy for breast cancer.

Objective:To discuss the new idea of on-the-spot recompression treatment and multidisciplinary treatment (MDT) for patients with unstable vital signs of type II decompression sickness. To provide reference for the nearby treatment of patients with critical decompression sickness.Methods:The clinical data of a case of a multi-disciplinary collaborative treatment of type II decompression sickness complicated with multiple organ dysfunction syndrome (MODS) admitted to a third-class A hospital in January 2020 were analyzed and summarized.Results:The patient suffered from consciousness disturbance and shock after 3 min of diver's blow-up out of the water. CT examination showed gas accumulation in the systemic multi-organ venous system, and laboratory examination suggested MODS. The oxygen inhalation regimen was given in the session of recompression treatmen by 0.12-0.18 MPa. Intravenous fluid was the total of 8900 ml in the session, and the total recompression treatment time was 9 h 45 min. The patient was still in unconscious when he finished the session. CT re-examination confirmed the elimination of venous bubbles, and laboratory examination indicated multiple organ failure (MOF) . The patient was given comprehensive supporting treatment by mechanical assisted breathing and following by continuons renal replacement therapy (CRRT) and extrocorporeal membrane oxygenation (ECMO) in the intensive care unit, and was discharged after 32 d of hospitalization.Conclusion:Critical decompression sickness patients with unstable vital signs are taken to a local general hospital with hyperbaric oxygen chamber and intensive care unit. The successful treatment can be achieved by organizing diving medicine, hyperbaric oxygen medicine and critical medical personnel for MDT.

Objective:To discuss the new idea of on-the-spot recompression treatment and multidisciplinary treatment (MDT) for patients with unstable vital signs of type II decompression sickness. To provide reference for the nearby treatment of patients with critical decompression sickness.Methods:The clinical data of a case of a multi-disciplinary collaborative treatment of type II decompression sickness complicated with multiple organ dysfunction syndrome (MODS) admitted to a third-class A hospital in January 2020 were analyzed and summarized.Results:The patient suffered from consciousness disturbance and shock after 3 min of diver's blow-up out of the water. CT examination showed gas accumulation in the systemic multi-organ venous system, and laboratory examination suggested MODS. The oxygen inhalation regimen was given in the session of recompression treatmen by 0.12-0.18 MPa. Intravenous fluid was the total of 8900 ml in the session, and the total recompression treatment time was 9 h 45 min. The patient was still in unconscious when he finished the session. CT re-examination confirmed the elimination of venous bubbles, and laboratory examination indicated multiple organ failure (MOF) . The patient was given comprehensive supporting treatment by mechanical assisted breathing and following by continuons renal replacement therapy (CRRT) and extrocorporeal membrane oxygenation (ECMO) in the intensive care unit, and was discharged after 32 d of hospitalization.Conclusion:Critical decompression sickness patients with unstable vital signs are taken to a local general hospital with hyperbaric oxygen chamber and intensive care unit. The successful treatment can be achieved by organizing diving medicine, hyperbaric oxygen medicine and critical medical personnel for MDT.

Cas1-and-Cas2-mediated new spacer acquisition is an essential process for bacterial adaptive immunity. The process is critical for the ecology of the oral microflora and oral health. Although molecular mechanisms for spacer acquisition are known, it has never been established if this process is associated with the morphological changes of bacteria. In this study, we demonstrated a novel Cas2-induced filamentation phenotype in E. coli that was regulated by co-expression of the Cas1 protein. A 30 amino acid motif at the carboxyl terminus of Cas2 is necessary for this function. By imaging analysis, we provided evidence to argue that Cas-induced filamentation is a step coupled with new spacer acquisition during which filaments are characterised by polyploidy with asymmetric cell division. This work may open new opportunities to investigate the adaptive immune response and microbial balance for oral health.

PURPOSE: Numerous previous studies have reported inconsistent results about the differences between synchronous contralateral breast cancer (sCBC) and metachronous contralateral breast cancer (mCBC). This study aimed to compare the clinical characteristics and outcomes between sCBC and mCBC and determine predictive factors for the survival of sCBC and mCBC patients. METHODS: Using the Surveillance, Epidemiology, and End Results Program database, we identified sCBC or mCBC patients from 2000 to 2010. The Kaplan-Meier method and Cox proportional hazards regression analysis were used to analyze overall survival and breast cancer-specific survival (BCSS) rates of sCBCs and mCBCs, respectively. RESULTS: Overall, 14,057 sCBC (n = 8,139, 57.9%) and mCBC (n = 5,918, 42.1%) patients were included. The first tumors of sCBC were more likely to have higher stage and more lymph and distant metastases, whereas those of mCBC were more often infiltrating ductal carcinoma (IDC), had localized stage, were estrogen receptor (ER) and progesterone receptor (PR) negative, and had less axillary nodal involvement. The second tumors of mCBC tended to be IDC and have higher grade, adverse stage, ER and PR-negativity; and more axillary nodal involvement, compared to the second tumors of sCBC. mCBC patients had significantly favorable 5-year BCSS but worse long-term BCSS compared with sCBC patients. Moreover, subgroup analysis revealed no significant difference of BCSS between sCBC and mCBC among patients aged 18–60 years. Multivariate analysis indicated that age, grade, and stage of 2 tumors; surgery for second tumor; and ER status of the second tumor were independent prognostic factors for BCSS of contralateral breast cancer (CBC). CONCLUSION: The characteristics and outcomes of sCBCs and mCBCs were substantially different. sCBC and mCBC patients may have different prognosis, and the prognosis of CBC depends on the first and second tumors.
Age of Onset ; Breast Neoplasms ; Breast ; Carcinoma, Ductal ; Estrogens ; Humans ; Methods ; Multivariate Analysis ; Neoplasm Metastasis ; Prognosis ; Receptors, Progesterone ; Risk Factors ; SEER Program

OBJECTIVE To analyze the data of non-invasive prenatal testing based on specific loci of circulating cell-free fetal DNA (cffDNA). METHODS Selected loci of target chromosomes were analyzed by sequence capture and sequencing. Meanwhile, 600 loci were selected from other chromosomes for determining the concentration of cffDNA. RESULTS A total of 768 specific loci were captured on chromosomes 21 and 18, and used to determine whether the two were abnormal. When the minimum concentration of detected cffDNA was set at 3% and the threshold of Z score was set to [-6,6], the specificity of the analysis was 99.37% and the sensitivity was 100%. CONCLUSION A reliable, convenient and low-cost analytical method has been developed. The method requires less sequencing data for non-invasive prenatal testing, and can accurately detect abnormalities of fetal chromosomes 21 and 18, and simultaneously determine the concentration of cffDNA.

Objective To establish a noninvasive prenatal testing(NIPT) method based on the chromosome specific site sequencing instead of the conventional whole genome sequencing.Methods Blood plasma samples from 200 pregnant women with known fetus karyotypes were collected.First,the specific loci on chromosome 13,18 and 21 were selected by the database filter.Then,these specific loci were captured by probes,and were performed sequencing.Finally,chromosome aneuploidy was identified by the Z-score.ResultsSeven fetuses with 21-trisomy syndrome,3 with 18-trisomy syndrome and 1 with 13-trisomy syndrome were detected by the chromosome specific site sequencing,and the results were accordant with those of the whole genome sequencing method.Conclusion The NIPT method based on the chromosome specific site sequencing has the advantages of low cost and high throughput,which may replace the NIPT method based on the whole genome sequencing.