ObjectiveTo explore the mechanism of the anti-cancer compound formula Feiyanning in inhibiting epithelial-mesenchymal transition （EMT） and invasion and metastasis of non-small cell lung cancer （NSCLC）. MethodsCell proliferation and activity were assessed using the cell counting kit-8（CCK-8） assay to evaluate the effect of Feiyanning on the proliferation of A549 and H1299 cells. Wound healing and Transwell assays were conducted to examine Feiyanning's impact on the metastasis of A549 and H1299 cells. The effects of Feiyanning on EMT and the transforming growth factor-β₁ （TGF-β₁）/Smad signaling pathway proteins in A549 and H1299 cells were detected by Western blot. Exogenous TGF-β₁ was used to induce EMT in A549 and H1299 cells. The effects of Feiyanning on TGF-β₁-induced NSCLC cell metastasis， EMT， and the TGF-β₁/Smad pathway proteins were assessed by wound healing assay， Transwell assay， and Western blot. In vivo， an A549 lung metastasis model was established via tail vein injection in nude mice. A total of 28 SPF male nude mice were randomly divided into four groups： Model （NC） group， Feiyanning low-dose （FYN1） group， Feiyanning high-dose （FYN2） group， and the positive control group （TGF-β receptor kinase inhibitor SB431542 group）. The corresponding interventions were performed. After 40 days， the mice were euthanized， and lung metastases were analyzed. The expression of E-cadherin， N-cadherin， p-Smad2， and p-Smad3 in each group was detected by immunohistochemistry （IHC）. ResultsAfter Feiyanning intervention， compared to the blank group， Feiyanning inhibited the proliferation of A549 and H1299 cells in a concentration-dependent manner （P<0.01）. The metastasis ability of Feiyanning-treated cells was significantly decreased compared to the blank group （P<0.01）. The expression of EMT marker proteins N-cadherin and zinc finger transcription factors （Zeb1， Snail， Slug） was significantly reduced in the Feiyanning groups compared to the blank group （P<0.05， P<0.01）. The expression of p-Smad2/3， Smad2/3， TβRI， and TβRⅡ， key proteins in the TGF-β₁/Smad signaling pathway， was also significantly decreased （P<0.01）. In the TGF-β₁-induced EMT model， compared to the TGF-β₁ group， the cell metastasis ability in the Feiyanning groups was reduced （P<0.01）， and the expression levels of N-cadherin， Zeb1， Snail， and Slug were significantly lower （P<0.01）. The expression levels of p-Smad2/3， Smad2/3， TβRI， and TβRⅡ were also significantly reduced （P<0.01）. In vivo results showed that compared to the model group， the number of lung metastases in the FYN1， FYN2， and SB431542 groups was reduced （P<0.01）， and the range of cell infiltration was narrowed. Immunohistochemical results showed that compared to the model group， the expression of E-cadherin in the FYN1， FYN2， and SB431542 groups was increased （P<0.01）， the expression of N-cadherin decreased （P<0.05， P<0.01）， and the expression of p-Smad2 and p-Smad3， key proteins of the TGF-β₁/Smad pathway， was reduced （P<0.01）. ConclusionFeiyanning inhibits the invasion and metastasis of NSCLC cells and EMT. The mechanism is related to the inhibition of TGF-β₁/Smad signaling pathway.

ObjectiveTo explore the mechanism of the anti-cancer compound formula Feiyanning in inhibiting epithelial-mesenchymal transition （EMT） and invasion and metastasis of non-small cell lung cancer （NSCLC）. MethodsCell proliferation and activity were assessed using the cell counting kit-8（CCK-8） assay to evaluate the effect of Feiyanning on the proliferation of A549 and H1299 cells. Wound healing and Transwell assays were conducted to examine Feiyanning's impact on the metastasis of A549 and H1299 cells. The effects of Feiyanning on EMT and the transforming growth factor-β₁ （TGF-β₁）/Smad signaling pathway proteins in A549 and H1299 cells were detected by Western blot. Exogenous TGF-β₁ was used to induce EMT in A549 and H1299 cells. The effects of Feiyanning on TGF-β₁-induced NSCLC cell metastasis， EMT， and the TGF-β₁/Smad pathway proteins were assessed by wound healing assay， Transwell assay， and Western blot. In vivo， an A549 lung metastasis model was established via tail vein injection in nude mice. A total of 28 SPF male nude mice were randomly divided into four groups： Model （NC） group， Feiyanning low-dose （FYN1） group， Feiyanning high-dose （FYN2） group， and the positive control group （TGF-β receptor kinase inhibitor SB431542 group）. The corresponding interventions were performed. After 40 days， the mice were euthanized， and lung metastases were analyzed. The expression of E-cadherin， N-cadherin， p-Smad2， and p-Smad3 in each group was detected by immunohistochemistry （IHC）. ResultsAfter Feiyanning intervention， compared to the blank group， Feiyanning inhibited the proliferation of A549 and H1299 cells in a concentration-dependent manner （P<0.01）. The metastasis ability of Feiyanning-treated cells was significantly decreased compared to the blank group （P<0.01）. The expression of EMT marker proteins N-cadherin and zinc finger transcription factors （Zeb1， Snail， Slug） was significantly reduced in the Feiyanning groups compared to the blank group （P<0.05， P<0.01）. The expression of p-Smad2/3， Smad2/3， TβRI， and TβRⅡ， key proteins in the TGF-β₁/Smad signaling pathway， was also significantly decreased （P<0.01）. In the TGF-β₁-induced EMT model， compared to the TGF-β₁ group， the cell metastasis ability in the Feiyanning groups was reduced （P<0.01）， and the expression levels of N-cadherin， Zeb1， Snail， and Slug were significantly lower （P<0.01）. The expression levels of p-Smad2/3， Smad2/3， TβRI， and TβRⅡ were also significantly reduced （P<0.01）. In vivo results showed that compared to the model group， the number of lung metastases in the FYN1， FYN2， and SB431542 groups was reduced （P<0.01）， and the range of cell infiltration was narrowed. Immunohistochemical results showed that compared to the model group， the expression of E-cadherin in the FYN1， FYN2， and SB431542 groups was increased （P<0.01）， the expression of N-cadherin decreased （P<0.05， P<0.01）， and the expression of p-Smad2 and p-Smad3， key proteins of the TGF-β₁/Smad pathway， was reduced （P<0.01）. ConclusionFeiyanning inhibits the invasion and metastasis of NSCLC cells and EMT. The mechanism is related to the inhibition of TGF-β₁/Smad signaling pathway.

Objective:To investigate the clinical effects of free posterior interosseous artery perforator flap on reconstruction of the soft tissue defects of multiple fingers in one hand.Methods:Clinical data of 9 patients with soft tissue defect of multiple fingers in one hand admitted to the Department of Hand Surgery, Yongkang Orthopedic Hospital between January 2021 and August 2023 were retrospectively studied. The patients were 7 males and 2 females, aged between 19 and 55 years old. The soft tissue defects of 2 patients with 3 fingers injury and 7 patients with 2 fingers injury were reconstructed with free posterior interosseous artery perforator flaps. The size of defects in single finger was 1.5 cm×2.5 cm-3.0 cm×4.5 cm. The size of flap was 2.5 cm×7.0 cm-3.5 cm×13.0 cm. The posterior cutaneous nerve of the forearm were sutured to the intrinsic nerve of palmar side of the 14 fingers. Donor sites in forearm were directly sutured. After discharge, regular follow-up at outpatient clinic and through WeChat interviews were conducted to observe the appearance and texture of the flaps, finger fullness, donor site function and appearance, as well as patient satisfaction. For the flaps with anastomosis of cutaneous nerve, the recovery of TPD of the flap was tested. Sensory recovery was assessed according to the British Medical Research Council (BMRC) sensory function assessment criteria. Hand functions were evaluated using the Total Active Movement (TAM) of fingers of the Hand Surgery of the Chinese Medical Association.Results:All of the 20 flaps survived and were evaluated through follow-up visits, which ranged from 6 to 25 months with an average of 13 months. The flaps were soft, unbloated and without obvious pigmentation. The protective sensations were recovered with an average static TPD of 14 flaps anastomosed with cutaneous nerve was of 9.7 (7-13) mm. Among them, S 3+ 10 fingers, S 3 4 fingers; 6 fingers of flap without sutured cutaneous nerve, S 2 5 fingers, S 1 1 finger. According to the TAM of Hand Surgery of Chinese Medical Association, the results were excellent for 15 fingers and good for 5 fingers. There was no scar contracture in the forearm donor sites and motor function was not affected. Conclusion:Posterior interosseous artery perforator flap has the advantages of thin flap, similar texture to the hand, good recovery in sensory, and minimal damage to the donor site. Therefore, it is a good option in reconstruction of soft tissue defects of multiple fingers in one hand.

Objective:To investigate the curative effect of transfer of free anteriolateral thigh flap (ALTF) with partial iliotibial tract on reconstruction of composite tissue defects in dorsal wrist/hand.Methods:Ten patients who were admitted in the Department of Hand Surgery of Yongkang Orthopaedic Hospital from March 2018 to August 2022 were entered in the study due to extensive composite tissue defects in dorsal wrists and hands. The patients were 24-48 (37 in average) years old. All patients had tendon defects in 3 or more digits with 35 digits in total. The length of the tendon defects were 5.0-11.0 cm in various digits. The soft tissue defects ranged from 6.0 cm×8.0 cm to 11.0 cm×21.0 cm in size. Six patients had defects accompanied with metacarpal and phalangeal bone fracture and joint capsule. In emergency surgery, all patients received thorough debridement and VSD coverage after internal fixation for combined fractures. At 3-5 days after the primary surgery, free ALTF transfer with partial iliotibial tract was employed to reconstruct the defects on dorsal wrists and hands and the digit extension function. The size of flap was 6.0 cm×9.0 cm-9.0 cm×17.0 cm, and the rest of defect wound was covered by skin graft. All the donor sites were sutured directly, except 1 that received abdominal skin graft. Patients had the follow-ups in outpatient clinics, or via telephone or Wechat, to observe the sensation and appearance of the flap, and the healing of the donor site. The recovery of the extension function of the hand was evaluated according to the totle active motion (TAM) system.Results:All flaps survived except one that had vascular compromise after the surgery and it survived after two explorations and a vascular transfer. Postoperative follow-ups lasted for 6 to 24 months, with 13 months in average. At the last follow-up, a all the flaps were found soft in texture and slightly swollen in appearance. There was no obvious pigmentation or rupture of skin and extensor tendon. Sensory recovery achieved to S 2 in 7 patients and S 3 in 3 patients. The hand function was evaluated according to TAM system and rated as excellent for 3 digits, good for 18 digits, fair for 9 digits and poor for 5 digits. There were linear scars in the donor sites. Conclusion:Free ALTF transfer with partial iliotibial tract in the reconstruction of composite tissue defect of dorsal wrists and hands can not only reconstruct the wound surface, but also the extension of digits, which cuts short the course of treatment.

Animals ; Coronary Vessels ; Diabetes Mellitus ; Muscle, Smooth, Vascular ; Myocytes, Smooth Muscle ; Rats

Objective:To investigate the demand and practical utility of simulation operations specialists (SOS) in simulation teaching modules during the standardized residency training.Methods:Based on the feedback for stimulated courses of standardized residency training, subjective evaluation of all residents, teachers and SOS who participated in simulation courses in 2017-2018 academic year were investigated and studied via the mobile phone online investigation. At the same time, the design data of teaching concept map of relevant curriculum were also included. The SPSS 13.0 was used to conduct the t test and chi-square test. Results:At present, only 26.3% of the preset functions were used in the medical simulation courses based on high-tech medical simulator. Tutors commanded less than 30% functions, while SOS participated in the whole process of the course preparation and commanded 63.6% of the course operations, which was higher than the requirement of teaching concept map (45.5%). Among them, ECG monitoring regulation, venous management and special effects makeup were in greatest needs and were items with the biggest gap between ideality and reality. Resident physicians required SOS to replace the tutors to operate teaching facilities, so as to reduce interruption (37.0%), implications (31.3%) during courses, and improvement of experience sense during the course (32.3%). Furthermore, specialists with clinical background needed more assistance from SOS than those without clinical background ( tQ3=3.204, tQ4=2.573, tQ5=2.660; P<0.05). Differences were found between the actual work content of SOS and their job requirement ( χ2=12.632, P<0.01). Conclusion:SOS plays a significant role in the simulation course of standardized residency training, especially in the course of clinical professional physicians. Auxiliary functions of simulated courses, such as teaching aids management, special effects makeup, course designing, qualified SP and others are the main necessities for SOS at present. Participation of tutors and SOS together is essential to ensure a good development and performance of medical simulation courses for standardized residency training.

Objective:To investigate the effects of osimertinib on proliferation, migration and invasion of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) overexpressing HCC827 cells and explore the potential mechanism of PLOD2 induced osimertinib resistance.Methods:We transfected HCC827 cells with LV-vector and LV-over/PLOD2. The expression of PLOD2 was detected by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. The effects of osimertinib on the proliferation of HCC827-vector and HCC827-PLOD2 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. The effects of osimertinib on the migration and invasion of HCC827-vector and HCC827-PLOD2 cells were determined by Transwell assays. The expressions of E-cadherin and vimentin in cells were detected by immunofluorescence (IF). The expressions of epithelial-mesenchymal transition (EMT), FAK-PI3K/AKT and MAPK signal pathway related proteins were detected by western blotting.Results:The MTT assay showed that HCC827-PLOD2 cells were hyposensitive to osimertinib. The 50% inhibitory concentration (IC 50) and resistance index of osimertinib for HCC827-PLOD2 cells was over 1 000 nmol/L and over 100, respectively. The result of wound healing assay showed that the migration distance of HCC827-PLOD2 was about (2.13±0.21) fold changes as that of HCC827-vector cells. The result of Transwell assay showed that the numbers of HCC827-PLOD2 passing through the matrix membrane were (212.78±10.43), significantly higher than (101.32±12.52) of HCC827-vector cells ( P<0.01). The result of IF showed that compared with HCC827-vector cells, the expression of E-cadherin was down-regulated while vimentin was up-regulated in HCC827-PLOD2 cells. Osimertinb downregulated E-cadherin and upregulated vimentin expression in HCC827-vector cells but had limited effect in HCC827-PLOD2 cells. The result of western blotting showed that PLOD2 significantly increased vimentin expression level while decreased E-cadherin expression level. Osimertinib inhibited the expression of p-EGFR, but did not affect the expressions of PLOD2, p-FAK, p-AKT, p-ERK, vimentin and E-cadherin in HCC827-PLOD2 cells. Conclusion:PLOD2 confers resistance to osimertinib in HCC827 cells by regulating EMT, FAK-PI3K/AKT and MAPK signal pathways.

Objective:To explore the clinical effect of repair of distal thumb degloving injury with B-shaped first palmar dorsal neurocutaneous vascular flap.Methods:The clinical data included 15 patients with distal thumb degloving injury received in Yongkang Orthopedic Hospital from March 2015 to June 2018. These patients included 10 males and 5 females, aged between 24 and 61 years. For the injury, 7 cases were beyond the interphalangeal joint, 8 cases were beyond the nail root. The length of the distal segment of degloving finger was 1.8 to 2.3 cm, and the skin and soft tissue defect ranged from 1.8 cm × 4.6 cm to 2.3 cm × 5.6 cm. The distal thumb degloving injury was repaired with B-shaped first palmar dorsal neurocutaneous vascular flap. The radial dorsal metacarpal nerve of the first metacarpal was anastomosed with the ulnar proper digital nerve stump. And full-thickness skin graft transplantation was performed in donor area. After the operation, the shape and function of the thumb were followed up. The sensory function of the skin was determined by British Medical Research Association Sensory Function Evaluation Standard, and the function of the thumb was evaluated with reference to the total active movement (TAM) of fingers of the Chinese Medical Association Hand Surgery Branch.Results:The flap area was 2.0 cm × 5.0 cm-2.5 cm × 6.0 cm, and all the flaps were survived. Follow-up period was 5-18 months after the operation, with an average of 10 months. The flap at the thumb repair site was soft, wear-resistant, non-bloated, no obvious pigmentation. Its protective sensation was restored. The two-point discrimination of the flap reached 8-11 mm, with an average of 9.3 mm, basically restoring the original shape and function of the thumb. There were 13 cases reaching the S3 + flap sensory function, 2 cases reaching S3. The thumb function was evaluated as excellent for 9 cases, and good for 6 cases. There were no complications such as scar contracture and hypersensitivity in the donor area. Conclusions:The repair of distal thumb degloving injury with B-shaped first palmar dorsal neurocutaneous vascular flap can complete skin coverage and sensory reconstruction, with satisfactory postoperative effect.

Objective:To investigate the effects of osimertinib on proliferation, migration and invasion of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) overexpressing HCC827 cells and explore the potential mechanism of PLOD2 induced osimertinib resistance.Methods:We transfected HCC827 cells with LV-vector and LV-over/PLOD2. The expression of PLOD2 was detected by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. The effects of osimertinib on the proliferation of HCC827-vector and HCC827-PLOD2 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. The effects of osimertinib on the migration and invasion of HCC827-vector and HCC827-PLOD2 cells were determined by Transwell assays. The expressions of E-cadherin and vimentin in cells were detected by immunofluorescence (IF). The expressions of epithelial-mesenchymal transition (EMT), FAK-PI3K/AKT and MAPK signal pathway related proteins were detected by western blotting.Results:The MTT assay showed that HCC827-PLOD2 cells were hyposensitive to osimertinib. The 50% inhibitory concentration (IC 50) and resistance index of osimertinib for HCC827-PLOD2 cells was over 1 000 nmol/L and over 100, respectively. The result of wound healing assay showed that the migration distance of HCC827-PLOD2 was about (2.13±0.21) fold changes as that of HCC827-vector cells. The result of Transwell assay showed that the numbers of HCC827-PLOD2 passing through the matrix membrane were (212.78±10.43), significantly higher than (101.32±12.52) of HCC827-vector cells ( P<0.01). The result of IF showed that compared with HCC827-vector cells, the expression of E-cadherin was down-regulated while vimentin was up-regulated in HCC827-PLOD2 cells. Osimertinb downregulated E-cadherin and upregulated vimentin expression in HCC827-vector cells but had limited effect in HCC827-PLOD2 cells. The result of western blotting showed that PLOD2 significantly increased vimentin expression level while decreased E-cadherin expression level. Osimertinib inhibited the expression of p-EGFR, but did not affect the expressions of PLOD2, p-FAK, p-AKT, p-ERK, vimentin and E-cadherin in HCC827-PLOD2 cells. Conclusion:PLOD2 confers resistance to osimertinib in HCC827 cells by regulating EMT, FAK-PI3K/AKT and MAPK signal pathways.

Objective:To explore the clinical effect of repair of distal thumb degloving injury with B-shaped first palmar dorsal neurocutaneous vascular flap.Methods:The clinical data included 15 patients with distal thumb degloving injury received in Yongkang Orthopedic Hospital from March 2015 to June 2018. These patients included 10 males and 5 females, aged between 24 and 61 years. For the injury, 7 cases were beyond the interphalangeal joint, 8 cases were beyond the nail root. The length of the distal segment of degloving finger was 1.8 to 2.3 cm, and the skin and soft tissue defect ranged from 1.8 cm × 4.6 cm to 2.3 cm × 5.6 cm. The distal thumb degloving injury was repaired with B-shaped first palmar dorsal neurocutaneous vascular flap. The radial dorsal metacarpal nerve of the first metacarpal was anastomosed with the ulnar proper digital nerve stump. And full-thickness skin graft transplantation was performed in donor area. After the operation, the shape and function of the thumb were followed up. The sensory function of the skin was determined by British Medical Research Association Sensory Function Evaluation Standard, and the function of the thumb was evaluated with reference to the total active movement (TAM) of fingers of the Chinese Medical Association Hand Surgery Branch.Results:The flap area was 2.0 cm × 5.0 cm-2.5 cm × 6.0 cm, and all the flaps were survived. Follow-up period was 5-18 months after the operation, with an average of 10 months. The flap at the thumb repair site was soft, wear-resistant, non-bloated, no obvious pigmentation. Its protective sensation was restored. The two-point discrimination of the flap reached 8-11 mm, with an average of 9.3 mm, basically restoring the original shape and function of the thumb. There were 13 cases reaching the S3 + flap sensory function, 2 cases reaching S3. The thumb function was evaluated as excellent for 9 cases, and good for 6 cases. There were no complications such as scar contracture and hypersensitivity in the donor area. Conclusions:The repair of distal thumb degloving injury with B-shaped first palmar dorsal neurocutaneous vascular flap can complete skin coverage and sensory reconstruction, with satisfactory postoperative effect.