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 assess the application value of CNVPLUS ?-array for the genetic analysis of spinal muscular atrophy (SMA). Methods:From June 2021 to December 2022, CNVPLUS ?-array technique was employed to test the SMN1 and SMN2 genes among peripheral blood samples from 17 suspected SMA patients, 18 core families with suspected SMA, and 25 healthy individuals. The results were compared with those of multiple ligation-dependent probe amplification (MLPA) assay. Samples with inconsistent results were subjected to nested PCR or comprehensive analysis of SMA. This study was approved by the Shanghai Institute for Pediatric Research, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (Ethics No. XHEC-D-2024-038). Results:CNVPLUS ?-array has identified 35 SMA patients, 36 carriers, and 25 healthy individuals. In comparison, MLPA has identified 34 SMA patients, 36 carriers, and 26 healthy individuals. The two methods demonstrated a high consistency ( Kappa = 0.968, P<0.001). Additionally, CNVPLUS ?-array has identified one patient with compound heterozygous variants of SMN1 and one carrier with a [2+ 0] genotype. Conclusion:CNVPLUS ?-array not only can accurately determine the copy numbers of SMN1 and SMN2 genes, but also identify point mutations in SMN1 and [2+ 0] carriers, which has offered a new method for the genetic testing of SMA.

To explore the value of CNVPLUS ?-array in the diagnosis of the DMD gene. A retrospective study was performed on 96 children who were clinically diagnosed with Duchenne or Becker muscular dystrophies(DMD/BMD) at the Department of Pediatric Endocrinology and Genetics of Xinhua Hospital affiliated to Shanghai Jiaotong University School of Medicine from January 2014 to March 2023. DNA was extracted from these children′s peripheral blood and divided into two parts. Variations of the DMD gene were detected by using CNVPLUS ?-array and sequential testing of MLPA—NGS—Sanger. In the sequential method, single exon deletions detected by MLPA were first verified by polymerase chain reaction (PCR) and then were tested by Sanger′s sequencing if PCR results were normal. The results showed that, among 96 samples, 91 cases with the pathogenic variation of the DMD gene were detected by the CNVPLUS ?-array, including 76 cases with large deletion/duplication (copy number variation, CNV) and 15 cases with small variation (single nucleotide variant or small insertion/deletion, SNV/Indel). All samples were tested and diagnosed within 5 days. In contrast, 76 cases with pathogenic CNV and 20 cases with pathogenic SNV/Indel were detected in the DMD gene by sequential method. However, all of the experiments and diagnoses were completed within 48 days. Moreover, 5 cases with SNV/Indel in the DMD gene were correctly clustered after the operation mode was optimized. In summary, as a new micro-array integrating CNV and SNV probes, CNVPLUS ?-array can detect CNV and SNV/Indel in the DMD gene simultaneously while the application of CNVPLUS ?-array could save a lot of time and manpower. CNVPLUS ?-array had an excellent diagnostic performance for CNV of the DMD gene. As for SNV/Indel, the diagnostic performance was slightly poor and the operation mode should be optimized. If necessary, other testing technologies should be supplemented to reduce the risk of missed diagnosis.

Objective:To evaluate the efficacy and safety of unilateral adrenalectomy for treating primary bilateral macronodular adrenal hyperplasia (PBMAH) of different clinical types.Methods:The clinical and biochemical data of 54 patients with PBMAH who underwent unilateral adrenalectomy from May 2008 to March 2023 were retrospectively collected. Preoperative CT images of all patients showed enlarged bilateral adrenal glands with multiple nodules of " fused masses". Mean preoperative blood cortisol concentration at 8am was (21.5±7.7)μg/dl, urinary free cortisol concentration was (442.6±300.4)μg/24h, and mean 8am ACTH concentration was (6.4±2.3)pg/ml. Postoperative symptoms, BMI, blood pressure, mass diameter, cortisol and ACTH concentration were recorded and analyzed.Results:Compared with ordinary laparoscopic surgery, robot-assisted surgery showed shorter operation time [(115.4±22.1)min vs.(95.0±19.8)min, P=0.045]; less blood loss [(118.2±57.0)ml vs. (125.6±45.3)ml, P=0.441] and shorter hospitalization time [(5.2±0.9)day vs. (6.4±1.2)day, P=0.279]. Compared with laparoscopic surgery, open surgery showed longer operation time [(134 34.5) min vs. (104.3±20.1) min, P=0.035]; more blood loss [(305.5±85.2) ml vs. (122.5±44.3) ml, P=0.012] and longer hospitalization time[(10.4±3.2)day vs. (5.7±1.0) day, P=0.020]. The average follow-up time was (23.7±11.7) months. Sixteen cases biochemically relapsed, and the average relapse-free time was (25.4±13.4) month. Mean postoperative systolic blood pressure was (131.1±16.8)mmHg ( P=0.001) while diastolic blood pressure decreased to (82.2±11.1)mmHg ( P=0.002). Postsurgical average blood cortisol concentration decreased to (10.2±4.0)μg/dl ( P<0.01), while urine cortisol concentration decreased to (106.6±43.4)μg/24h( P<0.01). Average ACTH concentration increased to (12.6±4.1)pg/ml( P=0.005). Recurrent patients had higher preoperative blood and urine cortisol concentration(24.7±8.2)μg/dl( P=0.046), (522.8±234.2)μg/24h( P=0.028), and all of them underwent contralateral adrenalectomy. Conclusions:Unilateral adrenalectomy is safe and effective for treatment of PBMAH while part of patients biochemically relapsed. Subclinical patients were observed no recurrent cases after surgery. Recurrent patients have higher preoperative blood and urine cortisol levels and should undertake contralateral adrenalectomy and supplement corticosteroids for whole life.

To explore the value of CNVPLUS ?-array in the diagnosis of the DMD gene. A retrospective study was performed on 96 children who were clinically diagnosed with Duchenne or Becker muscular dystrophies(DMD/BMD) at the Department of Pediatric Endocrinology and Genetics of Xinhua Hospital affiliated to Shanghai Jiaotong University School of Medicine from January 2014 to March 2023. DNA was extracted from these children′s peripheral blood and divided into two parts. Variations of the DMD gene were detected by using CNVPLUS ?-array and sequential testing of MLPA—NGS—Sanger. In the sequential method, single exon deletions detected by MLPA were first verified by polymerase chain reaction (PCR) and then were tested by Sanger′s sequencing if PCR results were normal. The results showed that, among 96 samples, 91 cases with the pathogenic variation of the DMD gene were detected by the CNVPLUS ?-array, including 76 cases with large deletion/duplication (copy number variation, CNV) and 15 cases with small variation (single nucleotide variant or small insertion/deletion, SNV/Indel). All samples were tested and diagnosed within 5 days. In contrast, 76 cases with pathogenic CNV and 20 cases with pathogenic SNV/Indel were detected in the DMD gene by sequential method. However, all of the experiments and diagnoses were completed within 48 days. Moreover, 5 cases with SNV/Indel in the DMD gene were correctly clustered after the operation mode was optimized. In summary, as a new micro-array integrating CNV and SNV probes, CNVPLUS ?-array can detect CNV and SNV/Indel in the DMD gene simultaneously while the application of CNVPLUS ?-array could save a lot of time and manpower. CNVPLUS ?-array had an excellent diagnostic performance for CNV of the DMD gene. As for SNV/Indel, the diagnostic performance was slightly poor and the operation mode should be optimized. If necessary, other testing technologies should be supplemented to reduce the risk of missed diagnosis.

Primary female urethral adenocarcinoma is rare. This paper reports a case of primary urethral mucinous adenocarcinoma complicated with signet ring cell carcinoma. The patient underwent urethral tumor resection in another hospital. Postoperative examination indicated that the tumor remained, and the tumor was completely removed after urethral tumor resection. After 11 months of follow-up, there was no tumor residue or recurrence.

Objective To discuss the clinical characteristics of adrenal metastases,and summarize the experience of diagnosis and treatment.Methods From January 2008 to June 2018,the clinical data of 55 patients with adrenal metastases treated in our hospital were analyzed retrospectively.This study included 34 male patients and 21 female patients and the median age was 60 years old (ranged 55 to 84 years old).The median value of maximum diameter of adrenal metastases was 3 cm (ranged 1.9 to 10.3 cm);with 35 cases on the left side,13 cases right and 7 cases bilateral.The primary sites of malignant tumors were pancreas (18 cases,32.7％),lung (12 cases,21.8％),liver (6 cases,10.9％) and colorectum (6 cases,10.9％),respectively.Thirty-four cases were confirmed by pathology after adrenalectomy and 21 cases were confirmed by needle biopsy.Thirty cases were diagnosed synchronously with the primary tumor and 25 cases were metachronous.The median time from diagnosis of primary tumors was 13.3 months (ranged 2.0 to 97.4 months).42 cases of these 55 cases were diagnosed within one year.Treatment options for adrenal metastatic lesions included single adrenalectomy in 18 cases,adrenalectomy combined with radiotherapy 16 cases,single intravenous chemotherapy 10 cases,intravenous chemotherapy combined with non-operative treatment 10 cases and single radiotherapy 1 case.Results The main pathological types were adenocarcinoma (19 cases,34.5％),ductal adenocarcinoma (10 cases,18.2％),hepatocellular carcinoma (6 cases,10.9％) and clear cell carcinoma (4 cases,7.3％).Two cases were lost follow-up and the follow-up rate was 96％,the median follow-up time was 8 months (ranged 1 to 135 months).The median overall survival (OS) time of 55 patients was 5.3 months (ranged 1 to 134 months).The one-year survival rate was 23.6％ (13/55),the two-year survival rate was 12.7％ (7/55),the three-year survival rate was 9.1％ (5/55) and the five-year survival rate was 1.8％ (1/55).Conclusions Pancreatic cancer was the most common type of malignant tumor for adrenal metastases in our hospital.Most primary tumors and adrenal metastases were diagnosed synchronously or within one year.Comprehensive treatment with retroperitoneal adrenalectomy may improve the OS,however the overall prognosis is poor.

Cerebral artery embolism is a rare but serious complication of facial plastic surgery. This paper reports a case of severe total cerebral arterial fat embolism caused by facial autologous fat injection. The patient past away after 40 hours of treatment. The cause of this total cerebral artery embolism patient was analyzed by tracing relevant medical history and cephalic CTA examination. By studying this case, we hope to reduce the occurrence of similar situations in the future.

BACKGROUND: A skull defect is inevitable after decompression treatment for traumatic brain injury. Titanium mesh as the most recognized skull repair material has good biocompatibility and has been widely used in clinical practice. However, the timing for skull repair after brain injury is still in dispute.OBJECTIVE: To compare the changes of brain perfusion and the recovery of neurological function in patients with skull defects before and after early and late-stage titanium mesh repair based on CT perfusion technique.METHODS: This was a single-center, prospective, observational clinical trial that was completed at the Taihe Hospital,Hubei University of Medicine in Hubei Province, China. Eighty-six patients with craniocerebral injury who had undergone decompression with removal of bone flap from January 2013 to January 2016 were recruited and subjected to skull repair using titanium mesh. All the patients were randomized into two groups: test group (n=40) with early skull repair within 1-3 months after decompression and control group (n=46) with late-stage skull repair within 6-12 months after decompression. CT perfusion technology was used to observe changes of brain perfusion at 3 days operatively and at 10 days postoperatively. The Barthel index was evaluated at 30 days postoperatively. The trial was registered with ClinicalTrial.gov (identifier: NCT03222297) on July 12th, 2017. The study protocol was approved by the Ethics Committee of Taihe Hospital with the approval No. 2012 (08), and performed in accordance with the Declaration of Helsinki,formulated by the World Health Organization and the hospital's ethical requirements for human research. All the patients and their families were voluntary to participate in the trial, were fully informed of the trial process, and then signed the informed consent prior to the initialization of the trial.RESULTS AND CONCLUSION: The postoperative cerebral blood volume and cerebral blood flow at the parietal cortex on the side of skull defect and at the cortex in the defect region were significantly higher in the two group than the baseline (P < 0.05), while the time to peak was lower than the baseline (P < 0.05). Compared with the control group,significantly higher cerebral blood volume and cerebral blood flow as well as shorter time to peak were observed in the test group (P < 0.05). The Barthel index of the test group was also significantly higher than that of the control group at 30 days postoepratively (P < 0.05). Overall, early skull repair with titanium mesh is helpful to improve the cerebral blood perfusion at the affected side and the recovery of neurological function. In addition, CT perfusion technology is a safe and effective method to monitor hemodynamic changes in the brain.