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 efficacy between robot-assisted and free-hand percutaneous cannulated screwing (PCS) in the treatment of femoral neck fractures in the middle-aged and young patients.Methods:A retrospective study was conducted to analyze the clinical data of 53 patients with femoral neck fracture who had been treated with PCS from May 2020 to May 2022 at Department of Traumatic Surgery, Maoming Hospital Affiliated to Southern Medical University. In the robot group of 25 patients subjected to robot-assisted PCS, there were 11 males and 14 females with an age of (48.2 ± 11.9) years; in the free-hand group of 28 patients subjected to free-hand PCS, there were 13 males and 15 females with an age of (48.5 ± 9.8) years. The 2 groups were compared in terms of operation time, intraoperative bleeding, intraoperative guide drills, intraoperative fluoroscopy frequency, fracture union time, Harris hip score at the last follow-up and postoperative complications. Postoperative imaging examination was performed to evaluate distribution accuracy of the cannulated screws in the femoral neck (deviation between the screws and the femoral neck axis, parallelism between the screws and distance between the screws and the neck cortex).Results:There was no statistically significant difference between the robot and free-hand groups in the general clinical data before operation, showing comparability ( P>0.05). The robot group showed significantly shorter operation time [(32.7 ± 4.8) min], significantly less intraoperative bleeding [(14.6 ± 4.8) mL], significantly less intraoperative guide drillings [(3.5 ± 0.7) times] and significantly less intraoperative fluoroscopy frequency [(7.9 ± 1.4) times] than the free-hand group [(56.9 ± 11.3) min, (27.0 ± 7.3) mL, (9.1 ± 1.8) times and (16.3 ± 6.0) times)] (all P<0.05). Postoperative imaging showed that the deviation between the screws and the femoral neck axis was 4.4° ± 1.1° on the anteroposterior X-ray film and 3.2° ± 0.8° on the lateral X-ray film, the parallelism between the screws 4.9° ± 0.8° on the anteroposterior X-ray film and 3.0° ± 0.7° on the lateral X-ray film, and the distance between the screws and the femoral neck cortex (10.4 ± 2.7) mm in the robot group, all significantly smaller than those in the free-hand group [10.5° ± 2.8°, 4.9° ± 1.1°, 12.1° ± 4.0°, 5.1° ± 1.3°, and (15.4 ± 3.2) mm] (all P<0.05). All the 53 patients were followed up for (22.2 ± 8.5) months. All fractures got united. The fracture union time in the robot group [(20.6 ± 4.6) weeks] was insignificantly shorter than that in the free-hand group [(23.7 ± 7.7) weeks] ( P>0.05). At the last follow-up, the Harris hip score in the robot group [(88.6 ± 5.6) points] was significantly higher than that in the free-hand group [(84.8 ± 6.3) points] ( P<0.05). Follow-ups revealed 2 cases of internal fixation loosening, 1 case of screw head cutting and 1 case of femoral head necrosis in the free-hand group but none of such complications in the robot group. Conclusion:In the treatment of femoral neck fractures in the middle-aged and young patients, compared with free-hand PCS, robot-assisted PCS shows advantages of shorter intraoperative time, less bleeding, less fluoroscopic radiation, higher accuracy of screw placement, a lower incidence of postoperative complications and better functional recovery of the hip joint.

Objective:To evaluate the follow-up clinical results of W-shaped acetabular angular plate for the treatment of acetabular posterior wall fractures through a direct posterior approach (DPA).Methods:Fifteen cases (10 males and 5 females, average aged 42.1±10.0 years) were involved in this study, with acetabular posterior wall fractures treated by using the W-shaped acetabular angular plate through a DPA between March 2017 and June 2019. Nine patients with fractures were injured by traffic accidents and the other 6 cases by falling. Based on the three-column classification for acetabular fractures, all of the cases belonged to posterior wall fractures (A2.1), which included 6 cases of simple fractures and 9 cases of comminuted fractures. The mean time interval between injury and surgery was 5.6±1.1 (range, 4-8) days. The DPA was adopted in all cases. The posterior wall fractures of the acetabulum were reduced and fixed with W-shaped acetabular angular plates. The reduction quality of the acetabulum was evaluated by X-ray and CT scan during follow-up visits according to the criteria proposed by Matta. The function of the hip joint was assessed by the Merle d'Aubigné-Posteal score modified by Matta.Results:The length of the surgical incision was 9.5±1.1 (range, 8-12) cm. The operation time was 45.3±10.1 (range, 35-75) mins. The amount of intraoperative blood loss was 248.0±94.7 (range, 100-380) ml. All the patients who were followed up for 20.5±6.3 (range, 16-38) months. All cases were evaluated according to Matta's reduction quality criteria, the satisfactory ratio of reduction was 100%. Among cases, 10 cases were matched the anatomic reduction, and the other 5 cases have belonged to satisfy. All cases of fractures had healed (the mean of healing time was 9.4±1.3 (range, 8-12 weeks). At the final follow-up visit, the mean of modified Merle d'Aubigne-Postel score was 16.9±1.6 (range, 13-18). Excellent clinical outcomes were obtained in 10 cases, good in 3 cases, and fair in 2 cases. One case had deep venous thrombosis of the lower extremities. The clots disappeared after anticoagulation treatment. One case had the heterotopic ossification, Brooker grade I, without any special treatment due to not affecting the hip joint activity in the follow-up visits. One patient had incision fat liquefaction and the wound healed after intensive dressing change. No internal fixation loosening or losing of fracture reduction was found at the follow-up visits.Conclusion:This study shows that using W-shaped acetabular angular plate for the treatment of acetabular posterior wall fractures through the DPA could obtain early satisfactory clinical outcomes.

Objective:To investigate the effect of indocyanine green angiography (ICGA) in the design of pre-expanded perforator flaps in the reconstruction of large defects.Methods:From October 2018 to October 2019, there were 22 patients undergoing facial and cervical reconstructive surgeries based on pre-expanded perforator flaps in the back region in Shanghai Ninth Peoples′ Hospital. 12 male and 10 female patients, ranging from 4 to 26 years old with an average age of 19, were all suffered from inferior facial and cervical defects after burns. Single-pedicled or multi-pedicled perforator flaps from back region were designed for treatment. ICGA was conducted after flap dissection intra-operatively to evaluate perfusion areas of perforators, which helped surgeons to determine whether supercharging should be conducted. After flap transplantation, ICGA was conducted again to evaluate the blood supply. Statistics of post-operative complications such as flap necrosis were collected.Results:the average flap size was 27 cm ×17 cm. With the guidance of ICGA, 15 cases remained the design of single pedicle including 5 superficial cervical artery (SCA) flaps and 10 free circumflex scapular artery (CSA) perforator flaps. Supercharging was performed in 7 cases: including 2 SCA flaps supercharged with CSA perforator and 5 free CSA perforator flaps supercharged with thoracodorsal artery (TDA) perforator. Venous thrombus occurred in 1 case but recovered after re-anastomosis of vessels. Except for 2 tip necroses, 20 flaps survived completely one week postoperatively. 2 tip necroses were recovered after dressing changes. All the patients were followed up for 5 to 16 months, with good flap color and texture, and improved function of head and neck.Conclusions:ICGA can be used as a safe and effective intraoperative detection method to guide the selection of appropriate perforators and the decisions on whether to conduct supercharging, which optimizes flap design and improves the success of surgery.

Objective:To investigate the effect of indocyanine green angiography (ICGA) in the design of pre-expanded perforator flaps in the reconstruction of large defects.Methods:From October 2018 to October 2019, there were 22 patients undergoing facial and cervical reconstructive surgeries based on pre-expanded perforator flaps in the back region in Shanghai Ninth Peoples′ Hospital. 12 male and 10 female patients, ranging from 4 to 26 years old with an average age of 19, were all suffered from inferior facial and cervical defects after burns. Single-pedicled or multi-pedicled perforator flaps from back region were designed for treatment. ICGA was conducted after flap dissection intra-operatively to evaluate perfusion areas of perforators, which helped surgeons to determine whether supercharging should be conducted. After flap transplantation, ICGA was conducted again to evaluate the blood supply. Statistics of post-operative complications such as flap necrosis were collected.Results:the average flap size was 27 cm ×17 cm. With the guidance of ICGA, 15 cases remained the design of single pedicle including 5 superficial cervical artery (SCA) flaps and 10 free circumflex scapular artery (CSA) perforator flaps. Supercharging was performed in 7 cases: including 2 SCA flaps supercharged with CSA perforator and 5 free CSA perforator flaps supercharged with thoracodorsal artery (TDA) perforator. Venous thrombus occurred in 1 case but recovered after re-anastomosis of vessels. Except for 2 tip necroses, 20 flaps survived completely one week postoperatively. 2 tip necroses were recovered after dressing changes. All the patients were followed up for 5 to 16 months, with good flap color and texture, and improved function of head and neck.Conclusions:ICGA can be used as a safe and effective intraoperative detection method to guide the selection of appropriate perforators and the decisions on whether to conduct supercharging, which optimizes flap design and improves the success of surgery.

Objective: To summarize clinical experience on reconstruction of severe facial disfigurement with flap prefabrication and soft tissue expansion. Methods: From September 2005 to June 2016, 49 patients with type Ⅲ and type Ⅳ facial deformities underwent facial reconstruction with an integrated method on the basis of prefabricated flaps. In the first stage, the descending branch of the lateral femoral circumflex vessels and the surrounding muscle fascia were dissected and transferred to subcutaneous pocket in the cervicothoracic area. The pedicles of the fascial flap were anastomosed to either the facial or superior thyroid artery and their venae comitantes in flap prefabrication. A tissue expander was placed beneath the fascial flap. In the second stage, over-expansion was achieved with intra-flap stem cell transplantation once patient′s skin showed signs of intolerance to expansion. In the third stage, prefabricated flap was transferred to cover the facial defects. the second or third internal mammary artery perforators or lateral thoracic artery perforators were reserved and flap supercharging would be performed depending on the perfusion of the flap revealed by indocyanine green angiography intra-operatively. Later, flap revisions further restored facial outline and delicate organ configuration. Aesthetic and functional status were independently graded to assess the facial appearance and function before and after the reconstruction. Results: 49 patients with severe facial deformities were included. 5 patients received stem cell transplantation. The final inflated volume ranged from 2 530 ml to 3500 ml and each patient had facial reconstruction with a prefabricated flap (range 23 cm×18 cm-34 cm×32 cm). Flap supercharging technique were used in 25 cases to augment blood perfusion, however, flap necrosis (5 cm× 2 cm) occurred in 1 patient, and tip necrosis occurred in 4 patients, otherwise, all flaps survived entirely. The aesthetic (1.15 to 2.29) and functional (0.86 to 2.42) status scores were statistically improved (P<0.01). Facial expressions such as smiling, blinking and frowning were noted. Conclusions Autologous full face reconstruction with an integrated method based on flap prefabrication can bring satisfying aesthetic and functional recovery, rendering a safe and effective option for most patients with massive facial defects.

Objective To investigate the effect of acute super volume blood dilution on cardiac output and blood coagulation function in senile patients.Methods Sixty patients with large joint operation were randomly divided into experimental group and control group,30 cases in each group.The experimental group was given 6％ hydroxyethyl starch and lactated Ringer's solution 15 mL/kg for AHH,while the control group was given routine infusion.Cardiac output and blood coagulation were compared between two groups.Results In the experimental group,cardiac output (CO) increased and blood coagulation function decreased after AHH (P ＞ 0.05).There were no significant differences in hematocrit (HCT),prothrombin time (PT),activated partial prothrombin time (APTT) and plasma fibrinogen (Fib) ateach time point between two groups (P ＞ 0.05).Conclusion Acute super volume blood dilution has small influence on cardiac output andblood coagulation function in senile patients.

Objective To investigate the effect of acute super volume blood dilution on cardiac output and blood coagulation function in senile patients.Methods Sixty patients with large joint operation were randomly divided into experimental group and control group,30 cases in each group.The experimental group was given 6％ hydroxyethyl starch and lactated Ringer's solution 15 mL/kg for AHH,while the control group was given routine infusion.Cardiac output and blood coagulation were compared between two groups.Results In the experimental group,cardiac output (CO) increased and blood coagulation function decreased after AHH (P ＞ 0.05).There were no significant differences in hematocrit (HCT),prothrombin time (PT),activated partial prothrombin time (APTT) and plasma fibrinogen (Fib) ateach time point between two groups (P ＞ 0.05).Conclusion Acute super volume blood dilution has small influence on cardiac output andblood coagulation function in senile patients.

Objective To observe the effect of the electroacupuncture ( EA) on the expression of cyclin-de-pendent kinase 5 ( Cdk5 ) in rats with muscle contusion and to explore its mechanism. Methods Thirty-two Sprague-Dawley rats were randomly divided into a normal group of 4, a model group of 4, a natural recovery group ( NR) of 12 and an EA group of 12. All except those in the normal group had acute skeletal muscle contusion induced through a heavy blow. The EA group was treated with 15 minutes of EA daily beginning 48 h after the injury while the other rats received no EA. The model group was sacrificed 24 h after modeling, and rats from the NR and EA groups were sacrificed on the 7th, 14th and 21st day after the modeling to collect tissues. Hematoxylin eosin ( HE) staining, Western blotting and quantitative real-time fluorescence PCR were used to observe any histological changes, as well as Cdk5 protein and mRNA expression. Results The HE staining showed that the other 3 groups displayed larger a-mounts of muscle fiber fracture, dissolution and inflammatory cell invasion than was observed in the normal group. Compared with the NR group, quicker recovery was seen in the EA group as evidenced by faster muscle satellite cell proliferation and more new muscle fiber generation. The average Cdk5 protein expression in both the NR and EA groups was higher than in the normal group, and that of the EA group was significantly lower than that of the NR group. Conclusions Muscle contusion can increase Cdk5 expression in skeletal muscles, at least in rats. EA can promote the restoration of skeletal muscle function, probably by inhibiting CDK5 protein and mRNA expression.

Objective] To study the effect of needle warming moxibustion and electroacupuncture in treatment of knee osteoarthritis. [Methods] 171 cases of patients with knee osteoarthritis were randomly divided into 3 groups: acupuncture and moxibustion group, electroacupuncture group, control group of 57 cases in each. The efficacy of the 3 groups was compared after 2 courses of post. [Results] Efficiency of the 3 groups of patients had no significant difference(P>0.05); The cure rate and significant efficiency rate of 3 groups were significantly different( P<0.05), it showed that the improvement of curative effect of each group was different;on the efficacy of the 3 groups through Kruskal-Wallis H, P<0.05, there was statistically significant difference among 3 groups. [Conclusion] The needle warming moxibustion has a best treatment effect and the electroacupuncture takes the second place. We should expand the use of acupuncture and moxibustion for the disease in clinic, the therapeutic effect of electroacupuncture has a gap with the needle warming moxibustion.