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 evaluate the efficacy of modified femoral neck osteotomy(mFNO)in the surgical treatment of patients with ankylosing spondylitis(AS)and severe spinal kyphosis combined with hip flexion contracture.Methods:A retrospective analysis was conducted on 61 AS patients(103 hips)with spinal kyphosis and hip flexion contracture who underwent pedicle subtraction osteotomy(PSO)and total hip arthroplasty(THA)from January 1,2019 to November 15,2023.Data on mFNO operation time,blood loss,preoperative and postoperative values of the angle of the trunk and lower limb(ATL),hip passive range of motion(ROM),visual analogue scale(VAS),and incidence of in-hospital compli-cations were recorded.Statistical analysis was performed using paired-samples t test.P＜0.05 was con-sidered statistically significant.Results:The study ultimately included 10 cases,9 males and 1 female,with an average age of(41.30±9.03)years.These patients underwent surgery for a total of 52 times,including 19 hips both receiving mFNO and THA,and 14 times PSO.The average operation time for nine bilateral mFNO was(133.11±34.81)min,with blood loss of(433.33±187.10)mL.A unilateral mFNO took 60 min with 200 mL of blood loss.The preoperative ATL of 19 hips was 40.37°±13.66°,and the postoperative ATL value was 88.47°±12.46°(P＜0.05).The preoperative VAS score was 0,while the postoperative VAS score was 5.95±1.51(P＜0.05).The preoperative hip extension ROM was 37.37°±18.13°,while the postoperative hip extension ROM was-4.95°±21.24°(P＜0.05).Hip flexion ROM improved from 37.37°±18.13° to 50.79°±20.36° after FNO(P＜0.05).There were three cases of in-hospital complications(3/52,5.67％):One case of postoperative atelectasis fol-lowing PSO(1/52,1.92％),one greater trochanter fracture identified during THA(1/52,1.92％),and one early dislocation post-THA(1/52,1.92％).Conclusion:mFNO significantly improves the ATL in AS patients with severe spinal kyphosis combined with hip flexion contracture,facilitating PSO and THA surgeries.

In order to provide basic information for the utilization and development of famous classical formulas containing Bletillae Rhizoma， this article systematically analyzes the historical evolution of the name， origin， harvesting and processing of Bletillae Rhizoma by reviewing the ancient materia medica， prescription books， medical books and modern literature. The research results showed that Baiji（白及） was the main name， some scholars took Baiji（白芨） as its main name， and there were many other names such as Baiji（白给）， Baigen（白根）， Baiji（白苙）. The mainstream source of Bletillae Rhizoma was the tubers of Bletilla striata， and drying， large， white， solid， root-free and skin removed completely were the good quality standards. With the promotion of wild to cultivated medicinal materials， there were certain differences between their traits， and the quality evaluation indexes should be adjusted accordingly. The origin of records in the past dynasties was widely distributed， with Guizhou and Sichuan having high production and good quality in modern times. The harvesting period is mostly in spring and autumn， and harvested in autumn was better. The processing and processing technology is relatively simple， and it was used fresh or powdered in past dynasties， while it is mainly sliced for raw use in modern times. Based on the results， it is suggested that the tubers of Bletilla striata of Orchidaceae should be used in the famous classical formulas， and it should be uniformly written as Baiji（白及）. And if the original formula indicates the requirement of processing， it should be operated according to the requirement， if the requirement of processing is not indicated， it can be used in raw form as medicine.

Objective:To investigate the mid-term clinical outcomes of selective column arthrodesis based on the three-column theory in the treatment of malunion of Lisfranc injury.Methods:The 28 patients with malunion of Lisfranc injury were analyzed retrospectively who had been treated by selective column arthrodesis at Department of Orthopedic Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine from January 2011 to January 2020.They were 18 males and 10 females, with an average age of 37.2 years(from 18 to 65 years). Twelve left and 16 right sides were affected. According to Myerson's three-column classification, one case was medial column injury (type A), 4 ones middle column injury (type B), 7 ones medial plus middle columns injury and 16 ones three-column injury. Medial column arthrodesis was conducted in 7, middle column arthrodesis in 4 and medial plus middle columns arthrodesis in 17. The American Orthopaedic Foot and Ankle Society (AOFAS) midfoot score and visual analogue scale (VAS) were compared between preoperation and the last follow-up to evaluate the improvements in foot function and pain. The operation-related complications were recorded.Results:All patients were followed up for an average of 35.6 months (from 18 to 60 months). The AOFAS midfoot score increased from 43.1±4.1 at pre-operation to 84.1± 7.4 at the last follow-up and the VAS score decreased from 5.7±1.3 at pre-operation to 2.0±0.9 at the last follow-up (both P<0.001). The wounds healed in 28 patients, 3 of whom had postoperative wound exudation but responded to dressing change. There were no such complications as injury to the deep peroneal nerve or deep venous thrombosis. The internal fixation was removed in 5 patients at about one year after arthrodesis. Conclusion:Selective column arthrodesis based on the three-column theory can result in satisfactory med-term clinical outcomes in the treatment of malunion of Lisfranc injury.

OBJECTIVE To prepare celastrol -loaded albumin nanoparticles （CLT-AN），and to investigate their activity against rheumatoid arthritis （RA）in vivo . METHODS CLT-AN was prepared by ultrasonic method . The formulation technology was optimized by single -factor test by taking particle size ，polydispersity index （PDI）and stability as indexes ，with the dosage of CLT ， the dosage of soybean oil and the ultrasonic power as factors . The physical and chemical properties of CLT -AN were investigated by transmission electron microscopy （TEM）and laser particle size analyzer ；in vitro stability and release profile were studied . A rat model of adjuvant -induced arthritis was constructed to investigate the effects of CLT -AN on joint swelling ，the levels of serum inflammatory factors [tumor necrosis factor α（TNF-α）and interleukin -1β（IL-1β）] and pathological state of joint tissue . RESULTS The optimized formulation was CLT 6.5 g，soybean oil 45 mg，ultrasonic power 490 W，ultrasonic time 8 min. CLT- AN prepared by the best formulation showed uniform and spherical morphology . Its particle size ，PDI，Zeta potential were （96.8± 1.1）nm，0.174±0.020，and（－18.6±1.7）mV，respectively. The encapsulation efficiency and drug -loading efficiency were （94.61±0.46）% and（2.42±0.21）%. There were no significant changes in particle size ，PDI，Zeta potential and encapsulation efficiency of CLT -AN within 5 days of storage at room temperature . CLT-AN was slowly released in vitro ，and the cumulative release reached 73.56% in 72 h. Compared with CLT ，CLT-AN could significantly inhibit the joint swelling of model rats ，reduced the levels of inflammatory factors TNF -α and IL -1β in serum ，and improved the pathological state of inflammatory joint tissue . CONCLUSIONS CLT-AN prepared by ultrasonic method has the appropriate particle size ，good stability ，significant sustained - release characteristics ，and excellent therapeutic efficacy against RA .

OBJECTIVE：To establish fingerprint of Huafengdan yaomu ，and to determine the contents of 7 nucleosides in samples of different fermentation time. METHODS ：HPLC method was adopted. The determination was performed on Pntulips BP-C18 column with mobile phase consisted of methanol-water （gradient elution ）at the flow rate of 0.8 mL/min. The detection wavelength was set at 260 nm，and column temperature was 35 ℃. The sample size was 10 μL. Using xanthine as reference， HPLC fingerprint of 12 batches of Huafengdan yaomu was drawn. The similarity of samples were evaluated with Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition）. Common peaks were confirmed. The contents of uracil ， hypoxanthine，xanthine，uridine，inosine，guanosine and thymidine were determined in samples of different fermentation time （0， 1，2，3，4 weeks）by the same method. RESULTS ：There were 8 common peaks in 12 batches of Huafengdan yaomu ，with similarities ranging from 0.712 to 0.954；7 components were identified ，namely uracil ，hypoxanthine，xanthine，uridine，inosine， guanosine and thymidine. The linear ranges of mass concentrations of above 7 components in samples at different fermentation time were 0.87-8.7 μ g/mL （r＝0.999 6）， 4030 1.51-15.1 μg/mL（r＝0.999 7），6.08-60.8 μg/mL（r＝0.999 5）， 号） 1.52-15.2 μg/mL（r＝0.999 6），1.82-18.2 μg/mL（r＝0.999 6）， 1.48-14.8 μg/mL（r＝0.999 6），1.63-16.3 μg/mL（r＝0.999 3）. The limits of quantification were 0.027 4，0.076 3，0.250 4，0.172 3，0.101 1，0.078 3，and 0.084 2 μ g/mL，and the detection limits were 0.008 7，0.025 5，0.007 9，0.084 1，0.035 7，0.026 9，0.027 5 μg/mL，respectively. RSDs of precision ， repeatability and stability tests （12 h）were all less than 3％. The sample recovery rates were 94.16％-100.16％（RSD＝2.24％，n＝ 6），93.87％-100.65％（RSD＝2.67％，n＝6），93.52％-99.66％（RSD＝2.30％，n＝6），93.67％-98.24％（RSD＝1.89％，n＝6）， 96.00％-102.18％（RSD＝1.96％，n＝6），94.62％-101.54％（RSD＝2.82％，n＝6），97.72％-104.56％（RSD＝2.97％，n＝6）. After fermentation for 0-4 weeks，the contents of the above 7 components and total nucleosides were 0.042-0.232，0.027-0.181， 0.039-0.651，0.026-0.225，0.034-0.111，0.009-0.124，0.079-0.099，0.647-1.292 mg/g，respectively. After fermentation for 1-4 weeks，the contents of uracil ，hypoxanthine，xanthine and total nucleosides were significantly increased ，compared with 0 week of fermentation；the contents of uridine ，inosine and guanosine were significantly lower than those in 0 weeks. CONCLUSIONS ：The established fingerprint has strong characteristics and simple to operate ，which can be used for the quality control of Huafengdan yaomu；the content determination method is accurate and reliable ，and can be used to simultaneously determine the contents of 7 active nucleosides ；the content of nucleosides in Huafengdan muyao is affected by fermentation time.

OBJECTIVE：To estab lish fingerprint of Duzhong butiansu pill s，analyze its chemical pattern recognition ，and determine the contents of 7 components in Duzhong butiansu pills ，so as to provide reference for the quality control of the preparation. METHODS ：HPLC method was adopted. The determination was performed on Pntulips BP-C 18 Plus column with 0.2％ phosphoric acid water-acetonitrile as mobile phase （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 330 nm，and column temperature was 35 ℃. The sample size was 20 μL. With paeonol as the reference，the HPLC fingerprints of 12 batches of Duzhong butiansu pills （S1-S12） were established with Similarity Evaluation System for TCM Chromatographic Fingerprint （2012 edition）； common peaks were determined and the similarity was evaluated. The chromatographic peaks were identified by comparing with the reference substance. SPSS 21.0 and SIMCA 13.0 software were used for cluster analysis and principal component analysis ，and 22 common peaks were evaluated. The contents of the identified components in 12 batches of samples was determined by the above HPLC method. RESULTS ：A total of 22 common peaks were identified in the HPLC fingerprint of 12 batches of Duzhong butiansu pills ，and the similarity was no loss than 0.960. There were 7 chemical components identified ，which were gallic acid （peak 1），chlorogenic acid （peak 3），liquiritoside（peak 6），hyperoside （peak 7），verbascoside（peak 8），icariin（peak 14）and paeonol （peak 15）. Among the 12 batches of samples ，S1，S3-S5，S7， S9 and S 11 were classified as one category ，S2，S10 and S 124Y091 were clustered into one category ，S6 was one category and S was one category. The 22 common peaks were divided into three principal components. The characteristic value （15.130） and contribution rate （68.775％） of principal component 1 were the largest ，and the score coefficients of peak 3（0.305）and peak 4（0.298）were the highest. Among 12 batches of samples，the cont ents of above 7 components were 18.196 231.951 3，0.000 6-0.049 4，0.234 8-0.415 9，0.039 5-0.079 1，0.053 5-0.249 3，0.000 5-0.000 8，0.646 4-1.146 9 mg/g，respectively. CONCLUSIONS：HPLC fingerprint of Duzhong butiansu pills is established successfully. Twelve batches of samples are clustered into 4 category. Peak 3（chlorogenic acid ）and peak 4（unknown）may be the important factors causing the difference of samples. The content of gallic acid is the highest among the 7 components.

Objective To observe the effects of Siweiyuganzi prescription on anti-peroxidation and blood lipid levels in experimental rats with hyperlipidemia. Methods Sixty male Sprague-Dawley (SD) rats were divided into normal control group, hyperlipidemia model group, Xuezhikang group, Siweiyuganzi prescription large, medium and small dose group according to the random number table method, with 10 rats in each group. The hyperlipidemia rat model was established by intragastric feeding with high fat emulsion everyday 10 mL·kg-1·d-1; normal saline 10 mL/kg was given to the normal control group, twice a day by intragastric feeding; 3 dosages of Siweiyuganzi suspended fluid 12.8, 6.4, 4.3 g·kg-1·d-1 intragastric administrations were given to Siweiyuganzi prescription large, medium and small dose groups respectively; Xuezhikang suspended fluid 0.3 g·kg-1·d-1 was given to Xuezhikang group intragastrically;the same volume of normal saline was given to hyperlipidemia model group. After 4 weeks, the level changes of blood lipid, serum superoxide dismutase (SOD), malonaldehyde (MDA), hydroxymethylglutaryl Coenzyme A (HMG-CoA) were observed. Results Compared to those in the normal control group, the levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), MDA, content and positive expression of HMG-CoA, alanine aminotransferase (ALT) were all higher in hyperlipidemia model group [TG (mmol/L): 6.59±0.72 vs. 4.32±0.36, TC (mmol/L): 7.10±0.25 vs. 5.98±0.40, LDL-C (mmol/L): 4.18±1.30 vs. 2.33±0.35, MDA (μmol/L): 26.05± 5.99 vs. 10.08±1.98, HMG-CoA content (ng/L): 54.60±2.90 vs. 48.73±3.09, HMG-CoA positive expression in liver tissue:(57.80±12.30)% vs. (22.00±4.92)%, ALT (U/L): 106.83±15.75 vs. 81.97±13.18]; SOD and high-density lipoprotein cholesterol (HDL-C) in hyperlipidemia model group were significantly decreased [SOD (kU/L): 295.47±37.51 vs. 345.13±19.76, HDL-C (mmol/L): 2.32±0.49 vs. 4.84±0.45, both P < 0.05]. Compared with the hyperlipidemia model group, the TG, TC, LDL-C, MDA, contents and positive expression of HMG-CoA in each group were significantly reduced, and the SOD and HDL-C were obviously increased, and the changes in the Siweiyuganzi high dose group were more significant than those of the Siweiyuganzi middle-and low-dose groups [TG (mmol/L): 4.70±0.46 vs. 5.40±0.31, 5.70±0.41, TC (mmol/L): 5.80±0.23 vs. 6.14±0.20, 6.56±0.32, LDL-C (mmol/L): 2.56±0.45 vs. 2.93±0.33, 3.28±0.32, HDL-C (mmol/L): 4.58±0.28 vs. 3.89±0.30, 3.59±0.08, SOD (kU/L): 381.45±20.68 vs. 360.60±30.16, 325.49±32.13, MDA (μmol/L): 16.98±5.39 vs. 17.89±5.37, 21.03±6.01, HMG-CoA content (ng/L): 50.58±0.77 vs. 52.16±0.66, 52.90±0.91, HMG-CoA positive expression in liver tissue: (27.90±6.03)% vs. (32.20±7.00)%, (43.00±8.39)%, all P < 0.05]. In the normal control group, there were positive cells scattered in the central vein area and loosely distributed around the portal area in the rat liver; in the hyperlipidemia model group, the positive cells were increased in the central vein area and the cells in relatively great number were seen around the portal area. While the positive cells in Xuezhikang group and in the high, medium and low dose Siweiyuganzi groups were decreased. Conclusion Siweiyuganzi prescription can regulate the levels of blood lipids, prevent and treat the lipid peroxidation caused by hyperlipidemia, and inhibit excessive expression of HMG-CoA in experimental rats with hyperlipidemia.

Objective To evaluate the curative effect of Huoxue-Xiaozhi capsule on non-alcoholic fatty liver disease (NAFLD). Methods A total of 100 patients with NAFLD who met the inclusion criteria were divided into 2 groups with 50 patients in each group by random number table method. The treatment group was treated with Huoxue-Xiaozhi capsule, while the control group was treated with Compound Methionine Choline Tablets. Both groups were treated for 8 weeks. The ALT, AST, γ-GT (γ- glutamyltranspeptidase ), ALP (alkaline phosphatase, alkaline phosphate) were detected by automatic blood biochemical analyzer. The serum high sensitivity-C reactive protein (hs-CRP) and Homocysteine (Hcy) were detected by supplementary detection method. The serum IL-18 was detected by ELISA method, and liver imaging was performed before and after treatment. The symptoms and signs were evaluated, and clinical efficacy was evaluated. Results The total effective rate was 90.0％ (45/50) in the treatment group and 74.0％ (37/50) in the control group. The difference between the two groups was statistically significant (Z=-2.328, P=0.020 ). After treatment, the serum hs-CRP (2.4 ± 2.9 mg/L vs. 3.6 ± 2.8 mg/L, t=2.105), Hcy (11.2 ± 5.5 μmol/L vs. 13.9 ± 6.4 μmol/L, t=2.262), IL-18 (690.6 ± 61.3 ng/L vs. 775.4 ± 60.5 ng/L, t=6.962) in the treatment group were significantly lower than those in the control group (P<0.05 or P<0.01). After treatment, the improvement of hypochondriac lump, sallow complexion, dark complexion and hypochondriac dull pain in the treatment group were superior to those in the control group (Z values were -2.563, -2.788, -2.780, -2.220 respectively, all Ps<0.05 ). The Serum ALT, AST,γ-GT and ALP levels were significantly lower than those of the control group (t values were 18.820, 19.811, 10.221 and 3.248 respectively, all Ps<0.001 ). Conclusions The Huoxue-Xiaozhi capsule can improve the liver function and reduce the levels of serum hs-CRP, HCY and IL-18, and its curative effect of NAFLD.

Objective: To investigate the feasibility of early monitoring doxorubicin (DOX)-induced cardiotoxicity by apoptosis molecular imaging of 2-(5-[¹⁸F]fluoro-pentyl)-2-methyl-malonic acid (¹⁸F-ML-10) PET/CT. Methods: Forty-seven BALB/c mice were randomly divided into the chemotherapy group (n=30) and the control group (n=17) according to the random number table. The mice in chemotherapy group were intraperitoneally injected with DOX (4 mg/kg) once a week for 3 weeks and mice in the control group were injected with the same amount of normal saline. All mice were subjected to ¹⁸F-fluorodeoxyglucose (FDG) and ¹⁸F-ML-10 PET/CT imaging at day 0, 2, 9, 16, and left ventricular ejection fraction (LVEF) was continuously monitored using cine cardiac MR (cine-CMR) imaging. The region of interest (ROI) was delineated on PET/CT images, and the maximum percentage activity of injection dose per gram of tissue (%ID/g) was calculated. The mice were sacrificed after imaging, and the heart tissue was taken for HE staining and TdT-mediated dUTP nick end labeling (TUNEL) assay. One-way analysis of variance, independent-samples t test and Pearson correlation analysis were used to analyze the data. Results: In the chemotherapy group, the myocardial ¹⁸F-FDG uptake on day 0, 2, 9, 16 were (63.3±14.5), (93.7±24.0), (153.6±20.6) and (135.8±32.5) %ID/g respectively, and ¹⁸F-ML-10 uptake were (0.09±0.02), (0.18±0.03), (0.22±0.04) and (0.55±0.12) %ID/g respectively. Compared with baseline (day 0), ¹⁸F-FDG and ¹⁸F-ML-10 uptake were significantly increased in the chemotherapy group at each time point after DOX administration(F=6.823, 20.848, both P<0.01). The myocardial ¹⁸F-ML-10 and ¹⁸F-FDG uptake were essentially unchanged at all time points in the control group(F=2.036, 1.155, both P>0.05). TUNEL and HE staining indicated that the cardiomyocytes in the chemotherapy group showed obvious apoptosis and vacuolization, and the apoptotic index (AI) was positively correlated with the ¹⁸F-ML-10 uptake (r=0.950, P<0.01). The cine-CMR imaging results showed that the LVEF in the chemotherapy group continued to decrease after DOX administration (F=4.507, P<0.05), and significant difference was identified at day 16 (t=2.980, P<0.05). There was a significant negative correlation between ¹⁸F-ML-10 uptake and LVEF (r=-0.709, P=0.01). Conclusions Both ¹⁸F-FDG and ¹⁸F-ML-10 PET/CT imaging can early assess DOX-induced cardiotoxicity in vivo. Given the high targeting specificity of ¹⁸F-ML-10, it may have a greater clinical transformation advantage over ¹⁸F-FDG in early assessment of cardiotoxicity.