ObjectiveTo obtain a construct chimeric antigen receptor macrophages （CAR-M） targeting disialoganglioside GD2 （GD2） and harboring the intracellular domain of macrophage-specific activating signaling adaptor protein DNAX-activating protein 12 （DAP12）. MethodsA CAR molecule targeting GD2 with the DAP12 intracellular domain as the signaling module ［Anti-GD2 CAR （DAP12）］ was designed， synthesized， and cloned into an adenoviral vector. This virus was used to infect PMA-differentiated THP-1 macrophages to construct green fluorescent protein （GFP）-labeled Anti-GD2 CAR THP-1 cells. CAR expression was confirmed using live-cell imaging. In vitro co-culture assays were performed to evaluate the phagocytic function of empty， Anti-GD2 CAR-M （CD3ζ）， and Anti-GD2 CAR-M （DAP12） groups against glioma cells with high （T98G） and low （U251） GD2 expression. ResultsThe CAR adenovirus efficiently transduced THP-1 macrophages， resulting in successful generation of Anti-GD2 CAR-M. Flow cytometry analysis following co-culture with glioma cells showed that compared to the empty group， the Anti-GD2 CAR-M （CD3ζ） group exhibited significantly enhanced phagocytosis （P<0.05）. Furthermore， compared to the Anti-GD2 CAR-M （CD3ζ） group， the Anti-GD2 CAR-M （DAP12） group demonstrated a further increase in phagocytic capacity specifically against GD2-high T98G cells （P<0.05）. ConclusionAnti-GD2 CAR-M （DAP12） cells are successfully established by transducing PMA-differentiated THP-1-M with recombinant Anti-GD2 CAR （DAP12） adenovirus. The engineered CAR-M can specifically target phagocytose glioma cells with high GD2 expression.

ObjectiveTo obtain stably transfected chimeric antigen receptor macrophages （CAR-M） targeting disialosylganglioside （GD2） based on the constitutive expression of myeloid differentiation factor 88 （MYD88）. MethodsAn Anti-GD2 CAR adenoviral vector incorporating the CD3ζ （cluster of differentiation 3 zeta chain）-MYD88 fusion construct was designed and constructed. THP-1-derived macrophages （THP-1M）， induced by phorbol 12-myristate 13-acetate （PMA） differentiation， were infected with this adenovirus to generate Anti-GD2 CAR THP-1 cells bearing a green fluorescent protein （GFP） tag. CAR expression was monitored using a live-cell imaging system. The phagocytic function of Empty CAR-M， Anti-GD2 CAR-M（CD3ζ）， and Anti-GD2 CAR-M（CD3ζ-MYD88） groups against glioma cells was evaluated in vitro through co-culture experiments， and their polarization status was also analyzed. ResultsCAR adenovirus successfully transfected THP-1 to obtain Anti-GD2 CAR-M. After co-culture with glioma cells， flow cytometry results showed that compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88） group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）. Compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88）CAR-M group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）， and the expression of inflammatory cytokines interleukin （IL）-6 and IL-12 was increased in this group （P<0.05）. ConclusionThis study successfully constructed Anti-GD2 CAR-M cells and verified their phagocytic activity， demonstrating that constitutive expression of MYD88 can promote inflammatory polarization of CAR-M. This provides a potential structural modification approach for the development of pro-inflammatory CAR-M.

Objective To investigate the surgical method and clinical outcome using the free anterolateral thigh flap with the medial gastrocnemius artery as the recipient artery for the repair of complex wounds in the lower leg. Methods A retrospective analysis was conducted on the clinical data of 11 patients with complex wounds in the lower leg admitted to Suzhou Ruihua Orthopedic Hospital from October 2020 to September 2022. All patients had extensive skin and subcutaneous soft tissue defects in the lower leg and received free anterolateral thigh flap repair with the medial gastrocnemius artery as the recipient artery. The skin of donor site was directly sutured. Results All 11 free skin flap survived in 11 patients, and both the donor and recipient sites healed in stage I. All patients were followed up after discharge, with a follow-up time of 12 to 35 months. The follow-up results at 12 months postoperatively showed good wound healing, good flap appearance and elasticity, restoration of normal knee and ankle joint function, and independent living ability. The donor site wounds healed well without significant scar hyperplasia. At the last follow-up, the flap sensory function was graded as S₃ in 2 patients and S₂ in 9 patients. The functional score of the affected limb was excellent in 5 patients and good in 6 patients, with an excellent and good rate of 100%. Conclusion The medial gastrocnemius artery has a suitable vessel caliber for anastomosis and a constant location. The use of the free anterolateral thigh flap with this artery as the recipient artery for the repair of severe wounds in the lower leg can effectively cover soft tissue defects and avoid injuring the residual main vessels in the lower leg, resulting in satisfactory outcomes.

Objective:To explore the clinical effects of free anterolateral thigh perforator flap pedicled with descending genicular artery in repairing wounds after lower leg limb-sparing surgery.Methods:A retrospective observational study was conducted. From January 2019 to June 2021, 12 patients with wounds after lower leg limb-sparing surgery who met the inclusion criteria were admitted to Suzhou Ruihua Orthopedic Hospital, including 6 males and 6 females, aged 17 to 74 years, with original wound area ranging from 17 cm×9 cm to 40 cm×15 cm. Five patients had infection in wounds. The wounds were all repaired by free anterolateral thigh perforator flap from contralateral thigh, with area of 18 cm×10 cm to 37 cm×9 cm. The artery of flap was anastomosed with the descending genicular artery, and the wounds in donor areas were sutured directly. Seven patients were transplanted with split-thickness skin grafts from the contralateral thigh to cover the remaining wounds that can not be covered by flap and the wounds in donor areas were covered with gauze. During the operation, the types of perforating branch carried by flap and the types of arteries and veins in recipient areas were recorded. The survival and occurrence of vascular crisis of flap, the survival of skin graft, the wound healing in donor and recipient areas, and the length of hospital stay after flap transplant surgery were recorded. During follow-up, the color and texture of flap, reinfection in lower leg, and fracture healing were recorded. At the last follow-up, the limb salvage function of patients was evaluated according to the functional evaluation criteria of Chen Zhongwei's amputated limb replantation.Results:The types of perforating branches carried by flaps were as follows: 6 cases of only carrying the descending branch of the lateral circumflex femoral artery, 3 cases of only carrying the oblique branch of the lateral circumflex femoral artery, and 3 cases of carrying the descending branch of the lateral circumflex femoral artery and oblique branch of the lateral circumflex femoral artery after internal pressurization anastomosis. The types of arteries in the recipient area of flap were as follows: one case of main trunk of the descending genicular artery, 8 cases of the saphenous branch of the descending genicular artery, and 3 cases of the articular branch of the descending genicular artery. The types of veins in the recipient area of flap were as follows: 8 cases of one accompanying vein of the descending genicular artery and one branch of the great saphenous vein, and 4 cases of two branches of the great saphenous vein. All the flaps survived without vascular crisis, and all the skin grafts also survived. The wounds in the donor and recipient areas were all healed. The length of hospital stay of patient after flap transplant surgery ranged from 13 to 79 days. During the follow-up of 6 to 23 months, the color and texture of flap were both good, with no infection in lower leg wound. Internal or external fixation were removed after fracture healing in 5 patients, and bone graft internal fixation was performed in 7 patients whose fractures were not healed after surgery and all the incisions healed without infection. At the last follow-up, the limb salvage effect of patients was evaluated as followings: excellent in 7 patients, good in 4 patients, and fair in one patient.Conclusions:Free anterolateral thigh flap pedicled with descending genicular artery can effectively repair the wounds after lower leg limb-sparing surgery and control infection with short length of hospital stay, while not increasing the risk of secondary injury of distal limb vessels. Thus, it can obtain satisfactory limb salvage effect which is worthy of clinical promotion.

To investigate the clinical efficacy of using the posterior tibial artery and peroneal artery perforator flaps to repair the heel wounds. Methods From January, 2011 to May, 2018, heel soft tissue de-fect caused by trauma in 18 cases were treated by posterior tibial artery and peroneal artery perforator flaps respec-tively. The posterior tibial artery perforator flap was used in 11 cases, and the peroneal artery perforator flap was used in 7 cases. The area of flaps ranged from 5.0 cm×3.0 cm to 11.0 cm×9.0 cm. The length of the vascular pedicle was from 10.0 cm to 16.0 cm.After operation, the patients were followed-up regularly.The time of wound healing, appear-ance and texture of the flap, and function of ankle joint were observed. Results After the operation, 13 flaps sur-vived uneventfully. The wound achieved primary healing. Partial necrosis occurred in the distal of posterior tibial artery perforator flap in 2 cases, and repaired by skin graft 1 or 2 months later.Marginal necrosis occurred in posterior tibial artery perforator flap in 2 cases and in peroneal artery perforator flap in 1 case. And scar healing occurred in these 3 cases finally.All the 18 patients were followed-up for 3 to 60 months, with an average of 10 months. Fracture healing time was from 3-6 months, with an average of 4 months. Flap was soft with satisfied appearance in 16 cases. Obvious scar formation occurred in 2 cases. There was no obvious scar contracture in donor sites. There was no obvi-ous limitation of the flexion and extension function of the ankle joint in 18 cases. According to the American Or-thopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score, there was excellent in 16 cases, and good in 2 cases. Conclusion As for the characteristics of the heel wound, it is a simple and practical method to use leg perforator flap to repair.The flap is based on a long vascular pedicle.And the clinical effect is satisfied.

Objective: To compare the clinical efficacy of icotinib and gefitinib in the treatment of advanced(stage Ⅳ) lung adenocarcinoma patients with epidermal growth factor receptor(EGFR) sensitive gene mutation. Methods: Fifty-four advanced(stage Ⅳ) lung adenocarcinoma patients with EGFR sensitive gene mutation were selected.According to the random number table method, the patients were divided into two groups, with 27 cases in each group.The icotinib group received icotinib hydrochloride targeted therapy, and the gefitinib group was orally given gefitinib.The clinical efficacy, quality of life score, adverse reactions, progression free survival(PFS) were compared between the two groups. Results: There were no statistically significant differences in clinical curative effect between the two groups[complete remission(0 cases vs.0 cases), partial remission(9 cases vs.8 cases), stable(12 cases vs.14 cases), progress disease(6 cases vs.5 cases), objective response rate(33.3% vs.29.6%), disease control rate(77.8% vs.81.5%), Z=1.060, χ²=0.143, 0.100, all P>0.05]. After treatment, the differences of body function, social function, psychological function, common symptoms and side effects, specific modules in the gefitinib group were not statistically significant compared with those before treatment(t=1.402, 1.199, 1.840, 1.860, 1.275, all P>0.05). The icitinib group had better body function, psychological function, common symptoms, side effects and specific modules than before treatment(t=2.525, 3.335, 4.477, 3.778, all P<0.05). The psychological function, common symptoms and side effects, specific module life quality scores in the icotinib group were (39.72±4.23)points, (38.84±4.67)points, (38.94±4.56)points, respectively, which were higher than (37.08±5.14)points, (35.48±5.02)points, (35.85±4.97)points in the gefitinib group(t=2.061, 2.546, 2.380, all P<0.05). The incidence rate of Ⅰ-Ⅱ grade adverse reactions between the two groups had no statistically significant difference (χ²=4.667, P>0.05). The incidence rate of Ⅲ-Ⅳ grade adverse reactions of the icotinib group was 7.4%, which was lower than 25.9% of the gefitinib group(χ²=9.000, P<0.05). The total incidence rate of adverse reactions of the icotinib group was 40.7%, which was significantly lower than 70.4% of the gefitinib group(χ²=25.694, P<0.05). There was no statistically significant difference in mean PFS between the two groups in the 19delete mutation of EGFR gene(t=0.795, P>0.05). The average PFS of the icotinib group under EGFR gene 21L858R mutation was (14.62±3.85)months, which was longer than (10.73±5.61)months of the gefitinib group(t=2.971, P<0.05). Conclusion Icotinib, gefitinib in the treatment of advanced(stage Ⅳ) lung adenocarcinoma patients with EGFR sensitive gene mutation has similar clinical effect, but icotinib has higher safety, better tolerability, and can significantly improve the quality of life of patients, prolong the EGFR mutation of the 21L858R gene under the survival of patients.