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 cholecystokinin octapeptide(CCK-8)on glutamate transporter 1(GLT-1)expression in hippocampal astrocytes induced by glutamate(Glu).Methods The mouse hippocampal astrocytes were isolated and the toxicity of CCK-8 at different concentrations on the mouse hippocampal astrocytes was detected.The cells were divided into control group,Glu group,Glu+0.1 μmol/L CCK-8 group,Glu+0.5 μmol/L CCK-8 group and Glu+1.0 μmol/L CCK-8 group.MTT assay was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.Biochemical kit was used to detect Glu content in the extracellular supernatant,and qRT-PCR was used to detect the mRNA expression of GLT-1 and glutamate/aspartate transporter(GLAST).The protein expressions of Caspase-3,Bcl-2,GLT-1 and GLAST were detected by Western blotting,and the expression of TNF-α in the cell supernatant was detected by ELISA.Results CCK-8 at different concentrations had no significant effect on the proliferation of mouse hippocampal astrocytes.Compared with the control group,the cell proliferation ability and the expression levels of Bcl-2 protein,GLT-1 and GLAST mRNA and protein in Glu group were significantly decreased(all P＜0.01),the apoptosis rate,extracellular Glu content,Caspase-3 protein expression level in cells and TNF-α level in cell supernatant were significantly increased(all P＜0.01);Compared with the Glu group,the cell proliferation a-bility and the expression levels of Bcl-2 protein,GLT-1 and GLAST mRNA and protein in the Glu+0.5 μmol/L CCK-8 group and Glu+1.0 μmol/L CCK-8 group were significantly increased(all P＜0.05),the apoptosis rate,extracellular Glu content,Caspase-3 protein expression level in cells and TNF-α level in cell supernatant were significantly decreased(all P＜0.01).Con-clusion CCK-8 can inhibit Glu-induced inflammatory response of astrocytes,promote the expression of GLT-1,reduce the con-centration of extracellular Glu,promote cell proliferation and inhibit apoptosis.

Objective:To explore the effectiveness of limb motor rehabilitation program based on patient health engagement (PHE) model in patients with hemorrhagic stroke, and to provide reference for the limb motor rehabilitation management of hemorrhagic stroke patients.Methods:Through literature review and Delphi expert correspondence, a limb motor rehabilitation program for hemorrhagic stroke patients based on the PHE model was constructed. A non-contemporaneous controlled study was conducted, 45 hemorrhagic stroke patients hospitalized in the Department of Neurosurgery of Shanxi Bethune Hospital from March to August 2022 were selected by convenience sampling method as the control group, and routine exercise rehabilitation measure was given, 45 hemorrhagic stroke patients from September 2022 to February 2023 were selected as the intervention group, a limb motor rehabilitation program based on PHE model was implemented on the basis of control group. The functional exercise compliance, limb motor function, daily activity ability, emotional and social dysfunction of patients in the two groups were observed before intervention, 1 and 3 months after intervention respectively.Results:A total of 85 patients with hemorrhagic stroke were included. There were 42 patients in the intervention group, 25 males and 17 females, aged (52.07 ± 9.91) years old, and 43 patients in the control group, 21 males and 22 females, aged (53.93 ± 10.52) years old. There were no significant differences in the functional exercise compliance, limb motor function, daily activity ability, emotional and social dysfunction of patients before intervention between the two groups. At 3 months after intervention, the functional exercise compliance score in the intervention group was (40.83 ± 7.92) points, higher than that in the control group (37.14 ± 6.44) points, and the difference was statistically significant ( t = 2.36, P<0.05). At 1 and 3 months after intervention, the scores of limb motor function and daily activity ability in the intervention group were (27.12 ± 6.74), (33.67 ± 6.54) points and (61.31 ± 6.72), (74.40 ± 8.71) points, which were higher than (24.91 ± 6.03), (27.02 ± 6.59) points and (52.33 ± 9.78), (60.12 ± 10.03) points of the control group, the differences were statistically significant ( t values were 2.06-7.01, all P<0.05), the scores of emotional and social dysfunction were (75.52 ± 22.09) and (58.33 ± 18.88) points, which were lower than (86.02 ± 23.04), (78.51 ± 21.67) points of the control group, and the differences were statistically significant ( t = - 2.14, - 4.57, both P<0.05). Conclusions:The limb motor rehabilitation program based on the PHE model could improve the exercise compliance of patients with hemorrhagic stroke, improve the limb motor function and daily activity ability of patients, alleviate negative emotions, and reduce the level of social dysfunction.

Bladder cancer(BC) ranks the first of genitourinary tumor in China and is one of the most common urological malignancies, in which 25%-30% of patients were diagnosed with muscle-invasive bladder cancer. Radical cystectomy combined with pelvic lymph node dissection is the standard procedure for treatment, which can effectively avoid tumor recurrence or distant metastasis as well as improve the prognosis of patients. However, some patients may not tolerate or refuse to undergo radical bladder surgery due to worry about high complication rate, high morbidity and poor postoperative quality of life. With the increasing understanding of bladder cancer heterogeneity and biological behavior, the treatment of bladder cancer has changed from a surgery-based treatment model to an individualized and comprehensive treatment model by multidisciplinary collaboration. The bladder-preserving treatment can achieve the same oncological prognosis as that of radical bladder surgery with a better quality of life of the patients, which has become a hot topic and focus of research in muscle-invasive bladder cancer treatment. This article reviewed the progress of research related to the comprehensive treatment of muscle-invasive bladder cancer with preservation of the bladder.

Objective To investigate the association between diabetic retinopathy ( DR) and atherosclerosis cardiovascular disease ( ASCVD) .Methods Clinical data of 654 patients with type 2 diabetic mellitus (T2DM) were collected from a cross sectional , population based survey on chronic diseases and risk factors which was conducted in Beijing Changping district from July 2010 to March 2011.Among 654 T2DM patients, there were 73 patients with ASCVD (ASCVD group) and 581 patients without ASCVD ( non-ASCVD group ) .The association between DR and ASCVD was analyzed .Results Patients with ASCVD had significantly older age [58.5(53.9,65.9) years], more female sex[52(71.2%)], higher proportion of ASCVD history [45(61.6%)], higher levels of PG 2 h[16.26(11.08,19.20) mmol/L], HbA1c[7.20(6.55,8.85)%], systolic pressure [151(133,165) mmHg(1 mmHg＝0.133 kPa)] and lower eGFR[87.2(75.0,103.0) ml· min-1· 1.73 m-2] than non-ASCVD patients[52.4(46.5,58.3) years, Z＝-5.86, P＝0.00; 307(52.8%),χ2＝-8.86, P＝0.00; 256(44.1%),χ2＝8.07, P＝0.01; 13.10(8.99,17.93) mmol/L, Z＝-2.35, P＝0.02; 6.70(6.00, 7.90)%, Z＝-3.33, P＝0.00; 143(131,158) mmHg, χ2＝-2.28, P＝0.02; 94.6(84.8,106.3) ml· min-1· 1.73 m-2, Z＝-3.47, P＝0.00].The trend to develop DR in ASCVD group was significantly higher than that in non-ASCVD group [19.2%(14/74) vs.8.3%(48/581), χ2＝9.01, P ＝0.00] .DR was an independent statistical indicator of the presence of ASCVD [ OR ( 95%CI ): 2.64 ( 1.37 -5.06 ), P ＝0.00 ] . Furthermore, when DR was divided into NPDR and PDR according to its severity , only PDR was significantly associated with incident ASCVD [OR(95%CI): 12.05 (2.63-55.12), P＝0.00].After adjusting for traditional ASCVD risk factors , such an association still existed , with the risk of having ASCVD increasing by 718%[ OR (95% CI): 8.18 ( 1.56 -42.81 ), P ＝0.01] .DR associates strongly with ASCVD in the Chinese population with T 2DM.Conclussion With the severity of DR increasing , the risk of ASCVD also grows.After adjustment for traditional risk factors , PDR is still associated with the risk of prevalent ASCVD.

Objective: The aim of this study was to investigate the effect of co-culture with AEC (amniotic epithelial cell) on the biological characteristics of AMSC (amniotic mesenchymal stem cell), and to investigate the roles of SDF-1/CXCR4 axis in the homing and migration of AMSC. Methods: AMSC andAEC were isolated from human amnion, and then cultured, amplified and identified, respectively. TheAMSC were divided into three groups:AEC co-cultured group, serum-free cultured group and serum cultured group.After culture for 24 h, 48 h, and 72 h, the proliferation viability ofAMSC was measured by CCK-8 assay and trypan blue staining; the expression of CXCR4 mRNAwas analyzed by flow cytometry and Real-time RT-PCR, and the migration ability ofAMSC in vitro was observed by migration assay. Results: Cell viability (48 h and 72 h) and survival rate in the co-culture and serum groups were higher than those in the serum-free group (all P<0.05). The mRNA and protein expressions of CXCR4 in AMSC of the co-culture and serum-free groups were significantly higher than those of the serum group (P<0.05). The migration ability of AMSC in the co-culture and serumfree groups, which increase with the SDF-1 (stromal cell derived factor-1) concentration gradient, were higher than that in the serum group (P<0.05). Conclusion: AMSC co-cultured with AEC still have the basic biological characteristics of MSC, and showed good growth activity. Co-culture withAEC can up-regulate CXCR4 onAMSC surfaces and enhance the migration ability ofAMSC in vitro.

Objective To investigate the clinical values of NT-proBNP in left ventricular enlargement(LVE) and left ventricular dysfunction(LVD) of the patients with essential hypertension(EH) to provide a diagnostic basis for their diagnosis .Methods Plas-ma NT-proBNP concentrations in 120 patients with EH and in 29 normal controls were measured ,then the echocardiography exami-nation was performed to determine the left ventricular ejection fraction ( LVEF) ,left ventricular end-diastolic diameter(LVEDD) , left atrium(LA) and left ventricular systolic diameter(LVSDD) .The correlation between plasma NT-proBNP with LVE and LVD was analyzed .The diagnostic accuracy of NT-proBNP was analyzed by using receiver operating characteristic (ROC) curve .Results There were no statistically significant differences in the aspects of age ,sex and serum creatinine between the EH group and con-trol group ,but plasma NT-proBNP level of the former was significantly higher than that of the latter .The NT-proBNP level in the patients with LVE was significantly higher than that in the patients with normal left ventricle (P<0 .05) .The NT-proBNP level in the LVD patients was significantly higher than that in the patients with normal left ventricular function (P<0 .05) .The plasma NT-p roBNP level was positively correlated with LA ,LVEDD and LVSDD(r=0 .518 ,0 .58 ,0 .48 ,P<0 .01) while negatively crrelated with LVEF(r= -0 .61 ,P<0 .01) .The ROC curve showed that when the NT-proBNP was set at 380 pg/mL ,the sensitivity and specificity of NT-proBNP for diagnosing LVE were 80 .6% and 72 .4% ;which for diagnosing LVD were 80 .8% and 77 .4% ,re-spectively .Conclusion NT-proBNP is closely correlated with multiple ultrasonic indicators reflecting the left ventricular function and its level can reliably reflect the left ventricular contraction function ,which can serve as the marker for screening LVE and LVD in the patients with EH .

Objective T o explore the crim inal characteristics of forensic psychiatry expertise in depression patients and schizophrenics w ith hom icide behavior. Methods A total of 40 depression (depressive episode) patients and 50 schizophrenics w ith hom icide behavior w ere random ly assigned into the study group and control group, respectively. D ata of dem ographic and crim inal characteristic of the tw o groups w ere collected by a self-designed questionnaire, and then w ere com pared. Results T here w ere no statis-tical differences in age, education level and career betw een study and control groups (P>0.05). C om pared w ith the control group, the victim s in the study group w ere m ainly the patient's children and parents, and m ost offenders had suicidal behavior after hom icide (70% ). In study group, the m otives of crim e w ere m ainly extended suicide and indirect suicide, and m ost offenders had attem pted suicide (85% ) and dim inished capacity of crim inal responsibility (70% ), w hich in control group had no capacity of crim inal responsibility (56% ). E xcept for crim inal site, there w ere statistical differences in other crim inal charac-teristics betw een tw o groups (P<0.05). Conclusion T here are different crim inal characteristics betw een de-pression patients and schizophrenics w ith hom icide behavior in forensic psychiatry, and these characteris-tics should be considered w hen these tw o diagnoses are distinguished in forensic psychiatry expertise.

Objective To discuss the views and significance of the "parallel" training mode, and provide a reference for the smooth development of the graduate students' standardized training work. Methods The self-designed questionnaire included rotation/training, clinical skills, scientific research, supporting conditions, and the rationality and prospect of the integrated training mode. We chose the graduate students of professional type classes of Grade 2014 and 2015 in a university affiliated hospital of Kunming, in a secret-based manner , and questionnaires were distributed and collected by stratified sampling and random sampling according to the proportion of 50％ of the total number of them. Excel database was used for data processing and analysis. Results A total of 200 questionnaires were distributed and 167 were recovered with an effective recovery rate of 83.5％. The research items involved in the investigation of the students that were more satisfied with the project were as follows:methods of integrated training mode being more reasonable (85.0％, n=142), improving clinical comprehensive ability (83.2％, n=139), accelerating the development of high-level clinical talents (67.7％, n=113), conducive to unify the clinical practice standards (55.1％, n=92), conducive to employment (48.5％, n=81), saving medical resources (44.3％, n=78). Meanwhile, the survey found that graduate students were not satisfied with the following items: research time (88.0％, n=147), arrangements for plans of rotate conversion (65.8％, n=110), monthly income (89.8％, n=150). Conclusion The integrated training mode can accelerate the development of high-level clinical talents, save medical resources, further improve the medical education system and better improve the clinical com-prehensive ability of medical students. However, it still needs to be further improved in terms of the arrange-ments for plans of rotate conversion of clinical departments, cultivation of scientific research ability, and salary, etc.

Objective To compare the effect of treating cricopharyngeal achalasia in stroke survivors using transnasal or transoral balloon dilatation.Methods Thirty stroke survivors with cricopharyngeal achalasia were randomly divided into a transnasal and a transoral balloon dilatation group (group N and group O),each of 15.Both groups were given routine swallowing rehabilitation training as well as the transnasal or transoral balloon dilatation.Their heart rate was monitored during the dilatation.Nasal bleeding,mucous membrane swelling and pain were also observed.Their swallowing function was evaluated using the Fujishima Ichiro swallowing efficacy score (FISE) and videofluoroscopy (VFSS) before and after the intervention.Results After the treatment,the average FISE and VFSS scores of both groups had improved significantly comnpared to before the treatment but there were no significant differences between the groups.During the treatment,the average heart rate of group O increased significantly less than that of group N.The treatment acceptance of group O was 98.2％,significantly higher than that of group N (80.1％).One case of mucosal bleeding was observed in group O,and laryngeal edema occurred significantly less often than in group N (9 cases vs.7).The average pain score was also significantly lower in group O.Conclusions Balloon dilatation facilitates swallowing among stroke survivors with cricopharyngeal achalasia.The transoral approach can help to reduce the occurrence of complications such as mucosal bleeding,laryngeal edema and pain,and has better patient acceptance.