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 compare the clinical outcomes and biomechanical characteristics of 1-, 2-, and 3-level pedicle subtraction osteotomy (PSO), and establish selection criteria based on preoperative radiographic parameters. Methods: Patients undergone PSO to treat ankylosing spondylitis from February 2009 to May 2019 in Sun Yat-sen Memorial Hospital of Sun Yat-sen University were enrolled. According to the quantity of osteotomy performed, the participants were divided into group A (1-level PSO, n = 24), group B (2-level PSO, n = 19), and group C (3-level PSO, n = 11). Clinical outcomes were assessed before surgery and at the final follow-up. Comparisons of the radiographic parameters and quality-of-life indicators were performed among and within these groups, and the selection criteria were established by regression. Finite element analysis was conducted to compare the biomechanical characteristics of the spine treated with different quantity of osteotomies under different working conditions. Results: Three-level PSO improved the sagittal parameters more significantly, but resulted in longer operative time and greater blood loss (p < 0.05). Greater stress was found in the proximal screws and proximal junction area of the vertebra in the model simulating 1-level PSO. Larger stress of screws and vertebra was observed at the distal end in the model simulating 3-level PSO. Conclusion Multilevel PSO works better for larger deformity correction than single-level PSO by allowing greater sagittal parameter correction and obtaining a better distribution of stress in the hardware construct, although with longer operation time and greater blood loss. Three-level osteotomy is recommended for the patients with preoperative of global kyphosis > 85.95°, T1 pelvic angle > 62.3°, sagittal vertical alignment > 299.55 mm, and pelvic tilt+ chin-brow vertical angle > 109.6°.

Objective: To compare the clinical outcomes and biomechanical characteristics of 1-, 2-, and 3-level pedicle subtraction osteotomy (PSO), and establish selection criteria based on preoperative radiographic parameters. Methods: Patients undergone PSO to treat ankylosing spondylitis from February 2009 to May 2019 in Sun Yat-sen Memorial Hospital of Sun Yat-sen University were enrolled. According to the quantity of osteotomy performed, the participants were divided into group A (1-level PSO, n = 24), group B (2-level PSO, n = 19), and group C (3-level PSO, n = 11). Clinical outcomes were assessed before surgery and at the final follow-up. Comparisons of the radiographic parameters and quality-of-life indicators were performed among and within these groups, and the selection criteria were established by regression. Finite element analysis was conducted to compare the biomechanical characteristics of the spine treated with different quantity of osteotomies under different working conditions. Results: Three-level PSO improved the sagittal parameters more significantly, but resulted in longer operative time and greater blood loss (p < 0.05). Greater stress was found in the proximal screws and proximal junction area of the vertebra in the model simulating 1-level PSO. Larger stress of screws and vertebra was observed at the distal end in the model simulating 3-level PSO. Conclusion Multilevel PSO works better for larger deformity correction than single-level PSO by allowing greater sagittal parameter correction and obtaining a better distribution of stress in the hardware construct, although with longer operation time and greater blood loss. Three-level osteotomy is recommended for the patients with preoperative of global kyphosis > 85.95°, T1 pelvic angle > 62.3°, sagittal vertical alignment > 299.55 mm, and pelvic tilt+ chin-brow vertical angle > 109.6°.

Objective: To compare the clinical outcomes and biomechanical characteristics of 1-, 2-, and 3-level pedicle subtraction osteotomy (PSO), and establish selection criteria based on preoperative radiographic parameters. Methods: Patients undergone PSO to treat ankylosing spondylitis from February 2009 to May 2019 in Sun Yat-sen Memorial Hospital of Sun Yat-sen University were enrolled. According to the quantity of osteotomy performed, the participants were divided into group A (1-level PSO, n = 24), group B (2-level PSO, n = 19), and group C (3-level PSO, n = 11). Clinical outcomes were assessed before surgery and at the final follow-up. Comparisons of the radiographic parameters and quality-of-life indicators were performed among and within these groups, and the selection criteria were established by regression. Finite element analysis was conducted to compare the biomechanical characteristics of the spine treated with different quantity of osteotomies under different working conditions. Results: Three-level PSO improved the sagittal parameters more significantly, but resulted in longer operative time and greater blood loss (p < 0.05). Greater stress was found in the proximal screws and proximal junction area of the vertebra in the model simulating 1-level PSO. Larger stress of screws and vertebra was observed at the distal end in the model simulating 3-level PSO. Conclusion Multilevel PSO works better for larger deformity correction than single-level PSO by allowing greater sagittal parameter correction and obtaining a better distribution of stress in the hardware construct, although with longer operation time and greater blood loss. Three-level osteotomy is recommended for the patients with preoperative of global kyphosis > 85.95°, T1 pelvic angle > 62.3°, sagittal vertical alignment > 299.55 mm, and pelvic tilt+ chin-brow vertical angle > 109.6°.

Objective: To compare the clinical outcomes and biomechanical characteristics of 1-, 2-, and 3-level pedicle subtraction osteotomy (PSO), and establish selection criteria based on preoperative radiographic parameters. Methods: Patients undergone PSO to treat ankylosing spondylitis from February 2009 to May 2019 in Sun Yat-sen Memorial Hospital of Sun Yat-sen University were enrolled. According to the quantity of osteotomy performed, the participants were divided into group A (1-level PSO, n = 24), group B (2-level PSO, n = 19), and group C (3-level PSO, n = 11). Clinical outcomes were assessed before surgery and at the final follow-up. Comparisons of the radiographic parameters and quality-of-life indicators were performed among and within these groups, and the selection criteria were established by regression. Finite element analysis was conducted to compare the biomechanical characteristics of the spine treated with different quantity of osteotomies under different working conditions. Results: Three-level PSO improved the sagittal parameters more significantly, but resulted in longer operative time and greater blood loss (p < 0.05). Greater stress was found in the proximal screws and proximal junction area of the vertebra in the model simulating 1-level PSO. Larger stress of screws and vertebra was observed at the distal end in the model simulating 3-level PSO. Conclusion Multilevel PSO works better for larger deformity correction than single-level PSO by allowing greater sagittal parameter correction and obtaining a better distribution of stress in the hardware construct, although with longer operation time and greater blood loss. Three-level osteotomy is recommended for the patients with preoperative of global kyphosis > 85.95°, T1 pelvic angle > 62.3°, sagittal vertical alignment > 299.55 mm, and pelvic tilt+ chin-brow vertical angle > 109.6°.

Objective: To compare the clinical outcomes and biomechanical characteristics of 1-, 2-, and 3-level pedicle subtraction osteotomy (PSO), and establish selection criteria based on preoperative radiographic parameters. Methods: Patients undergone PSO to treat ankylosing spondylitis from February 2009 to May 2019 in Sun Yat-sen Memorial Hospital of Sun Yat-sen University were enrolled. According to the quantity of osteotomy performed, the participants were divided into group A (1-level PSO, n = 24), group B (2-level PSO, n = 19), and group C (3-level PSO, n = 11). Clinical outcomes were assessed before surgery and at the final follow-up. Comparisons of the radiographic parameters and quality-of-life indicators were performed among and within these groups, and the selection criteria were established by regression. Finite element analysis was conducted to compare the biomechanical characteristics of the spine treated with different quantity of osteotomies under different working conditions. Results: Three-level PSO improved the sagittal parameters more significantly, but resulted in longer operative time and greater blood loss (p < 0.05). Greater stress was found in the proximal screws and proximal junction area of the vertebra in the model simulating 1-level PSO. Larger stress of screws and vertebra was observed at the distal end in the model simulating 3-level PSO. Conclusion Multilevel PSO works better for larger deformity correction than single-level PSO by allowing greater sagittal parameter correction and obtaining a better distribution of stress in the hardware construct, although with longer operation time and greater blood loss. Three-level osteotomy is recommended for the patients with preoperative of global kyphosis > 85.95°, T1 pelvic angle > 62.3°, sagittal vertical alignment > 299.55 mm, and pelvic tilt+ chin-brow vertical angle > 109.6°.

Objective To study the effects of diacerein and its metabolite rhein on plasma inflammatory cytokine level and expression of perirenal adipose tissue chemerin in type 2 diabetes mellitus (T2DM) rats and its role in regulating glucose and lipid metabolism.Methods We randomly divided 56 SD male rats into 5 groups: normal control group (Group A), T2DM group (Group B), pioglitazone group (Group C), diacerein group (Group D), and pioglitazone+diacerein group (Group E).Group A was fed with ordinary feed whereas the other groups were fed with high-fat diet.At the end of week 8, rats in Groups B, C, D and E were injected intraperitoneally with 30mg/kg of STZ solution to create the model.Those in Group A were injected with the same volume of sterile sodium citrate solution.OGTT examination was taken to screen the model rats at the end of week 10.The successful mode was chosen according to OGTT result.Then Group C was treated with pioglitazone 10mg/(kg·d) orally, Group D with diacerein 50mg/(kg·d), Group E with pioglitazone 10mg/(kg·d)+diacerein 50mg/(kg·d), and Group A and B were given the same volume of normal saline.The intervention lasted 4 weeks.At the end of experiment weeks 10 and 14, FBG, FINS, TC, TG, LDL-C, IL-1β, IL-6, and TNF-α were detected in the fasting rats with free access to water.After blood sample was taken at the end of week 14, all rats were killed and theperirenal adipose tissue was isolated, the expression of chemerin in perirenal adipose tissue was detected by Western blotting.Results At the end of week 10, FBG, FINS, TG, TC, LDL-C, IL-1β, IL-6, and TNF-α were higher in Groups B, C, D and E than in Group A while HDL-C was lower (all P<0.01).At the end of week 14, TC, TG, and LDL-C were lower in Groups C, D and E than in Group B but higher than Group A while HDL-C was lower than in Group A (all P<0.05).Group E had greater changes in glucose and lipid metabolism and inflammatory cytokine level than Groups C and D (P<0.05).Western blotting results showed that the expression of chemerin in perirenal adipose tissue increased higher in Group B than in Groups A, C, D and E (P<0.05).The expression of chemerin were also higher in Groups C and D than in Groups A and E (P<0.05), but there was no significant difference between Groups A and E.Conclusion Diacerein can regulate the metabolism of glucose and lipid, improve insulin resistance by reducing the expression of chemerin and the level of inflammatory cytokines.

Objective Recent studies have shown that inflammatory cytokines are involved in the occurrence and development of diabetes mellitus .The article aimed to investigate the effects of anti-inflammatory drug--diacerein on hepatic PPAR-γand GLUT-2 protein expression and its role in the regulation of glucose and lipid metabolism in rats with type 2 diabetes mellitus ( T2DM) . Methods 55 male SD rats were randomly divided into 4 groups:normal control group (n=10), T2DM group (n=15), pioglitazone intervention group(n=15), and diacerein treatment group(n=15) .Rats in normal control group were fed with normal diet , the other 3 groups were fed with high fat diet .At the end of 8th experi-ment week, rats in 3 groups fed with high fat diet were treated with intraperitoneal injection of 30mg/kg streptozotocin ( STZ) solution, while rats in normal control group were injected with the same volume of sterile sodium citrate solution .At the end of 10th week, OGTT modeling rats were screened .Rats in pioglitazone intervention group were treated with 10 mg/kg pioglitazone by intragastric administra-tion, rats in diacerein group was treated with 50mg/kg diacerein by intragastric administration , and rats in normal control group and T2DM group were given the same volume of normal saline .The intervention lasted 4 weeks.At the end of 8th, 10th and 14th week, the blood examination of glycolipid , FINS, IL-1βand liver function indexes was done on fasting rats .Fourteenth weeks later , after getting blood samples , all rats were sacrificed and liver tissues were isolated .Western blot was applied in the detection of PPAR γand immu-nohistochemistry was applied to detect GLUT-2 protein in livers. Results At the end of 8th week, the FBG level in pioglitazone in-tervention group increased compared with normal control group ( P<0.05) , and the levels of TC , TG, FINS, LDL-C, ALT, AST, IL-1βin the other 3 groups increased significantly compared with normal control group (P<0.05).At the end of 10th week, in comparison with normal control group , the levels of FBG , FINS, TC, TG, LDL-C, ALT, AST and IL-1βincreased significantly ( P<0.05) , while the levels of HDL-C and ISI decreased significantly (P<0.01).Moreover, the levels of FINS, TC, TG, LDL-C, ALT, AST and IL-1βat 10th weekend increased significantly compared with those at 8th weekend(P<0.05).At the end of 14th week, the FBG levels in T2DM group, pioglitazone intervention group and diacerein treatment group were higher than that of normal control group (P<0.01), while compared with T2DM group, the FBG level in pioglitazone intervention group decreased and the levels of HbA 1c, TC, TG, LDL-C and ALT in pioglitazone intervention group and diacerein treatment group decreased ( P<0.05) along with the increased ISI and HDL-C levels ( P<0.05) .Compared with the results at 10th weekend , the levels of FBG , HbA1c, LDL-C, ALT, AST in T2DM group in-creased (P<0.05) while the levels of HbA1c, TC, ALT, AST and IL-1βin pioglitazone intervention group and diacerein treatment group decreased significantly at 14th weekend(P<0.01).The expression level of PPAR-γin pioglitazone intervention group (0.91± 0.03) was significantly higher than normal control group (0.82±0.15), T2DM group(0.75±0.28) and diacerein treatment group (0.83± 0.34) ( P<0.01) , and no significant difference was found between diacerein treatment group and normal control group( P>0 .05) show-ing higher levels compared with T 2DM group ( P<0.01).At 14th weekend, the GLUT-2 expression levels in normal control group (0.209±0.023), pioglitazone intervention group (0.226±0.017) and diacerein treatment group (0.232±0.012) were higher than that of T2DM group (0.173±0.009,P<0.01);and the GLUT-2 expression levels in pioglitazone intervention group and diacerein treatment group were higher than that of normal control group (P<0.05).The expression level of liver PPAR-γwas in positive correlation with those of GLUT-2 protein, HDL-C, FINS, ISI ( r=0.815, 0.780, 0.747, P<0.01) and in negative correlation with those of FBG , HbA1c, TC, TG, AST, ALT, IL-1β(r=-0.465,-5.716,-0.615,-0.675,-0.617,-0.521,-4.827, P<0.05). Conclusion Diacerein can enhance liver PPAR-γand GLUT-2 expression levels and reduce the levels of IL-1β, HbA1c and blood lipid, thus im-prove insulin resistance in T 2DM rats.

Objective To compare the early stage immune reconstitution of high-and standardrisk Philadelphia chromosome-negative acute lymphoblastic leukemia (ALL) CR1 patients who had haploidentical blood and marrow stem cell transplantation (HBMT).Methods A total of 49 Ph-negative ALL CR1 patients who received HBMT and had complete early stage immune reconstitution data(+30,+60 and +90 d post transplantation) from Jan.2010 to Dec.2012 were enrolled.Immunophenotyping for B and T lymphocytes was performed post HBMT via flow cytometry.Fresh peripheral blood cells were stained with fluorochrome-labeled monoclonal antibodies against cluster of differentiation CD19,CD3,CD4,CD8,CD45RO,CD45RA and CD28.The early reconstitution of lymphocyte subsets,survival and prognosis between standard-risk group,high-risk adult group and high-risk children group were compared.Results There were no significant differences in all these T lymphocyte subsets among three groups at the three check points (P＞0.05).Moreover,at the same time,comparable outcome had been achieved between standard-risk group (n=18),high-risk adult group (n=16) and high-risk children group (n=15).There were no differences in 2-y relapse incidence (27.8％ vs 31.3％ vs 26.7％,P=0.957),2-y non-relapse mortality (11.1％ vs 0 vs 13.3％,P=0.185),2-y leukemia free survival (61.1％ vs 68.8％ vs 60.0％,P=0.834) and overall survival (77.8％ vs 68.8％ vs 60.0％,P=0.529) among the three groups.Incidence of grade Ⅱ-Ⅳ aGVHD was 44.4％ vs 12.5％ vs 46.7％(P=0.075) and incidence of cGVHD was 61.1％ vs 50.0％ vs 40.0％ (P=0.249).Conclusion Comparison of immune reconstitution at early stage may be a reasonable cause to explain that equivalent outcomes were observed among high-and standard-risk Ph-negative ALL CR1 patients after HBMT.

Benign biliary stricture is a challenging problem in hepatobiliary surgery. Benign biliary stricture is associated with major portal vein variation, which is not be found in literatures. A male patient with benign biliary stricture was admitted to the Chinese PLA General Hospital in March, 2010.The stricture was located in the hilar confluence with intrahepatic biliary dilation and hepatolithiasis. The result of computed tomography showed that the hilar biliary confluence was compressed by the left portal vein and right anterior portal vein. The patient was cured after receiving gallbladder interposition, choledocholithotomy and T tube drainage. We suggested that the benign hilar biliary stricture due to portal vein variation may be named as biliary nut-craker syndrome.