Objective To investigate the potential mechanism of tumor-derived exosome miR-3173-5p promoting the activation of cancer-associated fibroblasts(CAFs)and regulating the proliferation,invasion and apoptosis of non-small cell lung cancer(NSCLC)cells.Methods Exosome(Exo)of NSCLC cell A549 was extracted,the morphology of Exo was observed by transmission electron microscopy,and the expression of Exo marker protein was detected by Western blot.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the expression of miR-3173-5p in A549 cells and Exo.The binding of miR-3173-5p to F-box/WD-40 domain protein11(FBXW11)was predicted by Starbase database,and verified by luciferase reporter gene analysis.Human lung fibroblast cell line(MRC-5)was treated with Exo and Exo inhibitor GW4869 or NC inhibitor/mimic or miR-3173-5p inhibitor/mimic and/or FBXW11 overexpression/empty vector,NC group,Exo group,Exo+GW4869 group,Exo-NC inhibitor group,Exo-miR-3173-5p inhibitor group,Exo-NC mimic group,Exo-miR-3173-5p mimic group,Exo-miR-3173-5p mimic+Vector group and Exo-miR-3173-5p mimic+FBXW11 group were set up respectively.The level of CAFs marker protein[Matrix metalloproteinase-9(MMP-9),α-Smooth muscle aorta(α-SMA),Chemokine(C-X-C motif)ligand 12(CXCL12),Fibronectin,Vimentin,Interleukin-1β(IL-1β),Interleukin-6(IL-6),Interleukin-8(IL-8)]in cells of each group was detected by Western blot analysis,and the influence of exosome miR-3173-5p and FBXW11 expression on CAFs activation was analyzed.Exo and Exo-miR-3173-5P mimic were separated and the supernatant of MRC-5 cells incubated with Exo-FBXW11 was used as conditioned medium(CM)to culture A549 cells.The NC-CM group,Exo-CM group,Exo-miR-3173-5p mimic-CM group and Exo-FBXW11-CM group were set up respectively.MTT assay,Transwell assay and flow cytometry were used to detect the effects of CAFs activation on proliferation,invasion and apoptosis of NSCLC cells.Results Transmission electron microscopy showed that the diameter of exosomal vesicles derived from NSCLC cells was about 30～100 nm.Western blot analysis showed that all exosome labeled proteins were positive.The expression of miR-3173-5p(33.45±3.16)in NSCLC-derived exosomes was significantly higher than that in tumor cells(1.01±0.07),and the difference was statistically significant(t=1.263,P<0.001).MiR-3173-5p targets FBXW11 and inhibits its expression.Compared with NC group,CAFs marker protein levels in Exo group were significantly increased(t=12.214～24.908),the expression of CAFs marker protein in Exo+GW4869 group was significantly inhibited compared with Exo group(t=13.160～25.143),the differences were statistically significant(all P<0.01),respetively.Compared with Exo group,miR-3173-5p inhibitor inhibited EXO-induced CAFs marker protein expression(t=11.059～21.094),and miR-3173-5p mimic promoted the expression of CAFs marker protein(t=12.943～18.671),the differences were statistically significant(all P<0.01),respectively.Overexpression of FBXW11 reversed the induction of CAFs activation by miR-3173-5p mimic.Compared with the NC-CM group,the proliferation activity and invasion rate of A549 cells in the Exo-CM group were significantly enhanced,and the apoptosis rate was significantly inhibited(t=10.207,2.359,3.001),miR-3173-5p mimic enhanced the promoting effect of Exo-CM on proliferation and invasion of A549 cells and the inhibitory effect on apoptosis(t=9.399,3.438,3.208),while overexpression of FBXW11 can antagonize the promotion effect of Exo-CM on proliferation and invasion of A549 cells and the inhibition effect on apoptosis(t=18.868,7.070,9.813),the differences were statistically significant(all P<0.05),respectively.Conclusion Tumor-derived exosome miR-3173-5p promotes CAFs activation through targeted inhibition of FBXW11 expression,further promotes proliferation and invasion of NSCLC cells,inhibits cell apoptosis,and regulates the occurrence and development of NSCLC.

Objective To investigate the potential mechanism of tumor-derived exosome miR-3173-5p promoting the activation of cancer-associated fibroblasts(CAFs)and regulating the proliferation,invasion and apoptosis of non-small cell lung cancer(NSCLC)cells.Methods Exosome(Exo)of NSCLC cell A549 was extracted,the morphology of Exo was observed by transmission electron microscopy,and the expression of Exo marker protein was detected by Western blot.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the expression of miR-3173-5p in A549 cells and Exo.The binding of miR-3173-5p to F-box/WD-40 domain protein11(FBXW11)was predicted by Starbase database,and verified by luciferase reporter gene analysis.Human lung fibroblast cell line(MRC-5)was treated with Exo and Exo inhibitor GW4869 or NC inhibitor/mimic or miR-3173-5p inhibitor/mimic and/or FBXW11 overexpression/empty vector,NC group,Exo group,Exo+GW4869 group,Exo-NC inhibitor group,Exo-miR-3173-5p inhibitor group,Exo-NC mimic group,Exo-miR-3173-5p mimic group,Exo-miR-3173-5p mimic+Vector group and Exo-miR-3173-5p mimic+FBXW11 group were set up respectively.The level of CAFs marker protein[Matrix metalloproteinase-9(MMP-9),α-Smooth muscle aorta(α-SMA),Chemokine(C-X-C motif)ligand 12(CXCL12),Fibronectin,Vimentin,Interleukin-1β(IL-1β),Interleukin-6(IL-6),Interleukin-8(IL-8)]in cells of each group was detected by Western blot analysis,and the influence of exosome miR-3173-5p and FBXW11 expression on CAFs activation was analyzed.Exo and Exo-miR-3173-5P mimic were separated and the supernatant of MRC-5 cells incubated with Exo-FBXW11 was used as conditioned medium(CM)to culture A549 cells.The NC-CM group,Exo-CM group,Exo-miR-3173-5p mimic-CM group and Exo-FBXW11-CM group were set up respectively.MTT assay,Transwell assay and flow cytometry were used to detect the effects of CAFs activation on proliferation,invasion and apoptosis of NSCLC cells.Results Transmission electron microscopy showed that the diameter of exosomal vesicles derived from NSCLC cells was about 30～100 nm.Western blot analysis showed that all exosome labeled proteins were positive.The expression of miR-3173-5p(33.45±3.16)in NSCLC-derived exosomes was significantly higher than that in tumor cells(1.01±0.07),and the difference was statistically significant(t=1.263,P<0.001).MiR-3173-5p targets FBXW11 and inhibits its expression.Compared with NC group,CAFs marker protein levels in Exo group were significantly increased(t=12.214～24.908),the expression of CAFs marker protein in Exo+GW4869 group was significantly inhibited compared with Exo group(t=13.160～25.143),the differences were statistically significant(all P<0.01),respetively.Compared with Exo group,miR-3173-5p inhibitor inhibited EXO-induced CAFs marker protein expression(t=11.059～21.094),and miR-3173-5p mimic promoted the expression of CAFs marker protein(t=12.943～18.671),the differences were statistically significant(all P<0.01),respectively.Overexpression of FBXW11 reversed the induction of CAFs activation by miR-3173-5p mimic.Compared with the NC-CM group,the proliferation activity and invasion rate of A549 cells in the Exo-CM group were significantly enhanced,and the apoptosis rate was significantly inhibited(t=10.207,2.359,3.001),miR-3173-5p mimic enhanced the promoting effect of Exo-CM on proliferation and invasion of A549 cells and the inhibitory effect on apoptosis(t=9.399,3.438,3.208),while overexpression of FBXW11 can antagonize the promotion effect of Exo-CM on proliferation and invasion of A549 cells and the inhibition effect on apoptosis(t=18.868,7.070,9.813),the differences were statistically significant(all P<0.05),respectively.Conclusion Tumor-derived exosome miR-3173-5p promotes CAFs activation through targeted inhibition of FBXW11 expression,further promotes proliferation and invasion of NSCLC cells,inhibits cell apoptosis,and regulates the occurrence and development of NSCLC.

Objective To study the effects of different implantation angles of bi-leaflet mechanical heart valve(BMHV)on swirling flow in the aorta.Methods Based on the aortic CT images of a healthy volunteer,the effects of 4 different valve implantation angles(0°,45°,90° and 135°)on the aortic swirling flow under constant flow conditions were studied by computational simulation.Results The implantation of BMHV could seriously interfere with the aortic swirling flow,affecting the structure and helicity distributions of the swirling flow in ascending aorta,thus resulting in disturbed blood flows distal to the valve.The 135° implantation angle caused the most disruption to the swirling flow,leading to the largest areas of reversed swirling flows,while the 0° and 45° implantation angles caused relatively smaller damage to the swirling flow.The areas with low wall shear stress(<0.5 Pa)were the smallest when the implantation angle of BMHV was 0°.Conclusions At the implantation angle of 0°-45°,the disruption of BMHV to the swirling flow in ascending aorta was relatively small.Therefore,for different patients,the selection of implantation angle should be individualized according to the spatial geometry of their aorta(including the aortic sinuses),and the implantation angle can be determined between 0°-45°.

Objective:To investigate the clinical efficiency of gastrocnemius muscle flap combined with antibiotics loaded calcium sulfate in the treatment of postoperative infection and plate exposure of tibial plateau fracture in elderly patients.Methods:From January, 2015 to May, 2019, 21 elderly patients with postoperative infection and plate exposure of tibial plateau fracture were treated, including 14 males and 7 females with an average age of 72.6 years, ranging from 61 to 82 years. The average course of disease was 22.7 days, ranging from 6 to 91 days. The site of wound infection was at medial in 8 cases, lateral in 9 cases and bilateral in 4 cases. The size of wound ranged from 2.0 cm×3.5 cm to 5.0 cm×12.0 cm. All wounds were implanted with antibiotics loaded calcium sulfate and repaired by gastrocnemius muscle flap combined with skin graft after debridement. Muscle flap survival, wound healing, inflammatory index, fracture healing and knee joint function were recorded. The curative effect was evaluated by McKee infection treatment criteria and the knee joint function was evaluated by HSS scoring criteria.Results:All 21 muscle flaps survived. In 1 case, skin graft necrosis occurred in a small area, and the wound healed well after dressing change. One case had exudation which was clear and the bacterial culturing was negative, and the wound healed after 2 weeks of dressing change. The other incisions were healed in stage I, and the healing rate was 90%. All patients were followed-up for an average of 28.7(16-39) months. The redness and swelling occurred in 1 case without exudation after 2 months which disappeared after anti-infection treatment and didn't recur again. The infection recurred in 1 case after 5 months, and it which was controlled after debridement and plate removal. According to McKee criteria, 19 cases were cured, 1 improved and 1 recurred with an effective rate of 95.2%. The fracture healing time was from 3 to 7 months, with an average of 4.6 months. According to HSS scoring criteria, the knee joint function was excellent in 12 cases, good in 7 cases and moderate in 2 cases.Conclusion:After thorough debridement, gastrocnemius muscle flap combined with antibiotic loaded calcium sulfate can effectively control the infection, repair the wound, promote fracture union and restore limb function in the treatment of postoperative infection and plate exposure of tibial plateau fracture in elderly patients.

Objective:To compare the clinical results of different endovascular thrombus reduction techniques in the treatment of acute lower extremity arterial thrombosis.Methods:The clinical data of 96 consecutive patients with acute femoral popliteal arterial thrombosis who were treated with catheter directed thrombolysis (CDT) and mechanical thrombus aspiration system (PMT) between Jan 2016 and Dec 2018 at Cangzhou People's Hospital and Peking University People's Hospital were retrospectively analyzed.Results:Ninty-six patients underwent thrombolytic surgery,including 36 with CDT thrombolysis, 28 with AngioJet aspiration alone and 32 with Rotarex aspiration alone. Angiojet thrombus aspiration reduced thrombus rate by 89.3% (25/28) and clinical success rate by 92.8% (26/28).The thrombus reduction rate of Rotarex group was 87.5% (28/32), and the clinical success rate was 96.8% (31/32). In the CDT thrombolytic group, the thrombolytic reduction rate was 61.1% (22/36), including 8 patients who underwent thrombectomy and 6 patients with PMT, with a clinical success rate of 86.1% (31/36). The rate of distal arterial embolization, puncture point and local subcutaneous hematoma and vascular rupture was 21.4%, 10.7% and 2.1%, respectively. There were no amputation cases reported during a mean 13 months follow-up.The survival rate was 97.9%. The first-stage patency rate of 67.8%, while the second-stage artery patency rate of 85.7% during the follow-up.Conclusion:Compared with CDT, PMT has higher efficiency and lower complication rate in the treatment of acute lower extremity arterial thrombosis.

Objective:To examine the preliminary clinical efficacy of Chinese magnetic sphincter augmentation (MSA) in the treatment of gastroesophageal reflux disease (GERD).Methods:According to the enrollment criteria for the MSA developed by ShengJieKang Co. and Shanghai Chest Hospital (SS-MSA) clinical trial, a total of 19 GERD patients were treated with SS-MSA from August 2018 to January 2020 at Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University. The majority of registered cases were male patients with age of (32.2±7.3) years (range: 22 to 50 years), height of (170.7±6.2) cm (range: 160 to 179 cm) and weight of (65.2±10.3) kg (range: 47.5 to 90.0 kg). SS-MSA was implanted via laparoscopy. The major evaluation indexs of postoperative efficacy were the total time of acid exposure within 24 hours and the total number of reflux. Secondary efficacy indicators included: (1) evaluation of the average daily dose of proton pump inhibitor medications; (2) the score of GERD health related quality of life questionnaire (GERD-Q) before and after MSA implantation. Paired design t-test was used to evaluate the efficacy of the SS-MSA. Results:A total of 19 patients underwent SS-MSA surgery successfully. The history of the GERD were 19 (54) months ( M( QR)). The operation time was 63 (22) minutes and the in-hospital stay was 3 (2) days. No obvious surgical complications occurred. Postoperative adverse events included 14 cases with mild to moderate dysphagia exited after surgery, gradually eased within 1 to 3 months, 1 case with the removal of the device after 1 month of severe swallowing difficulties, 1 case of diarrhea. No corrosion, perforation, displacement occurred. The GERD-Q score (11.0(4.5) vs. 6(1.0), t=4.274, P=0.013), 24-hour acid exposure time (6.2(4.8)% vs. 0.1(0.9)%, t=5.814, P=0.004), and Demeester score (23.72(16.20) vs. 0.96(3.10), t=6.678, P=0.003) were significantly decreased 1 year after surgery( n=5). Proton pump inhibitor reuse rates were 6/18, 5/15, 3/10, and 1/5 in 1, 3, 6 and 12 months after the operation, respectively. Conclusions:SS-MSA implantation is feasible and safe with short hospital stay and rare perioperative complications. The preliminary results is good after 1 year follow-up. It could be expected to be an ideal substitutive for future GERD treatment.

Objective:To examine the preliminary clinical efficacy of Chinese magnetic sphincter augmentation (MSA) in the treatment of gastroesophageal reflux disease (GERD).Methods:According to the enrollment criteria for the MSA developed by ShengJieKang Co. and Shanghai Chest Hospital (SS-MSA) clinical trial, a total of 19 GERD patients were treated with SS-MSA from August 2018 to January 2020 at Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University. The majority of registered cases were male patients with age of (32.2±7.3) years (range: 22 to 50 years), height of (170.7±6.2) cm (range: 160 to 179 cm) and weight of (65.2±10.3) kg (range: 47.5 to 90.0 kg). SS-MSA was implanted via laparoscopy. The major evaluation indexs of postoperative efficacy were the total time of acid exposure within 24 hours and the total number of reflux. Secondary efficacy indicators included: (1) evaluation of the average daily dose of proton pump inhibitor medications; (2) the score of GERD health related quality of life questionnaire (GERD-Q) before and after MSA implantation. Paired design t-test was used to evaluate the efficacy of the SS-MSA. Results:A total of 19 patients underwent SS-MSA surgery successfully. The history of the GERD were 19 (54) months ( M( QR)). The operation time was 63 (22) minutes and the in-hospital stay was 3 (2) days. No obvious surgical complications occurred. Postoperative adverse events included 14 cases with mild to moderate dysphagia exited after surgery, gradually eased within 1 to 3 months, 1 case with the removal of the device after 1 month of severe swallowing difficulties, 1 case of diarrhea. No corrosion, perforation, displacement occurred. The GERD-Q score (11.0(4.5) vs. 6(1.0), t=4.274, P=0.013), 24-hour acid exposure time (6.2(4.8)% vs. 0.1(0.9)%, t=5.814, P=0.004), and Demeester score (23.72(16.20) vs. 0.96(3.10), t=6.678, P=0.003) were significantly decreased 1 year after surgery( n=5). Proton pump inhibitor reuse rates were 6/18, 5/15, 3/10, and 1/5 in 1, 3, 6 and 12 months after the operation, respectively. Conclusions:SS-MSA implantation is feasible and safe with short hospital stay and rare perioperative complications. The preliminary results is good after 1 year follow-up. It could be expected to be an ideal substitutive for future GERD treatment.

Objective: To evaluate the safety and effectiveness of esophageal replacement with ileocolon graft. Methods: Totally 34 cases of esophageal replacement with ileocolon graft from July 2015 to November 2017 at Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiaotong University were analyzed retrospectively, including 24 male and 10 female, aging from 7 to 72 years old. Esophageal replacement with ileocolon graft by right and/or middle colic artery as a blood supply using retrosternal route except one subcutaneous route. The primary esophageal disease, postoperative complication rate and quality of life were analyzed. Results: The overall postoperative complication rate was 23.5% (8/34), cervical anastomotic leakage rate of 5.9% (2/34), necrosis of colon graft of 5.9% (2/34). There were 3 patients experienced re-operation including 2 patients with colon graft necrosis and 1 patient with intestinal obstruction after ERC. One patient with colon graft necrosis died of septic shock after reoperation. Six cases of cervical esophago-jejunal anastomosis stenosis and 1 case of diarrhea occurred in the later time. All patients were followed up for a median time of 9 months (range: 1 to 28 months), 32 cases survived but 1 patient died until last follow-up by the end of December 2017. Conclusion Esophageal replacement with ileocolon graft by right and/or middle colic artery as a blood supply using retrosternal route was safe and effective.

Objective To scan protein expression profile of immune complexes (ICs) derived from the synovial tissue of the patients with rheumatoid arthritis (RA) based on liquid chromatography-tandem mass spectrometry (LC-MS).Methods The samples of synovial fluid were obtained from knee joints of the patients with RA and osteoarthritis (OA) used as control during therapeutic arthrocentesis in knee jiont at the Department of Orthopedics of Jinling Hospital,School of Medicine,Nanjing University.The protein expression profile of ICs was identified by enrichment strategy based on immunoprecipitation and LC-MS analysis.The value of fraction of total (FOT) was used to estimate protein abundance and screen the up-and down-regulated proteins.The function enrichment,interaction network and signal pathway of differential proteins were analyzed using softwares David and String.Results A total of 511 and 526 protein spots in ICs of RA and OA patients were identified respectively.Among them,170 proteins existed only in RA group.45 and 85 proteins in RA group were statistically up-and down-expressed compared with controls.Conclusion HSP90AA1,HSP70,HLAG,Thioredoxin,Annexin A2 and vitronectin may be involved in the pathogenesis of RA through different paths and possible to become promising diagnostic indicators or new therapeutic targets for RA.

Objective To investigate the application value of lung ultrasonic on severe high altitude pulmonary edema.Methods A prospective, single-blind, case-control study was conducted. Sixty patients with severe high altitude pulmonary edema admitted to Qinghai University Affiliated Hospital from February 2015 to May 2017 were enrolled. The patients were divided into 2500-3000 m group, 3000-3500 m group and 3500-4200 m group according to different altitudes,with 20 patients in each group. The acute physiology and chronic health evaluationⅡ(APACHEⅡ) score was recorded before and 12 hours and 24 hours after treatment. The arterial partial pressure of oxygen (PaO2) was determined by blood gas analysis, and the oxygenation index (PaO2/FiO2) was calculated. Bedside ultrasound scanning was used to determine B line number and pulmonary artery pressure (PAP), and B line score was calculated to reflect lung water content. The correlation between B line score and PaO2/FiO2, PAP and APACHEⅡ scores at each time point was analyzed by Pearson correlation analysis.Results None of 60 patients died or exited, all of them were enrolled in the final analysis. There was no significant difference in PaO2/FiO2, PAP, APACHEⅡ score or B line score among different altitudes groups (allP > 0.05). Repeated measurement variance analysis showed that the effects of different altitudes on PaO2/FiO2, PAP, APACHEⅡ score and B line score were not statistically significant (F value was 0.312, 0.014, 1.098, 0.236, andP value was 0.340, 0.791, 0.733, and 0.986, respectively). The PaO2/FiO2, PAP, APACHEⅡ score and B line score in all groups were improved obviously from 12 hours after treatment, and the improvements at 24 hours were more than those at 12 hours (allP < 0.05). Repeated measurement variance analysis showed that the effect at different time points on PaO2/FiO2, PAP, APACHEⅡ score and B line score was statistically significant (F value was 1844.270, 121.690, 1173.175, 19426.968, allP < 0.001). The interaction effects of different altitudes and different time points on PaO2/FiO2, PAP, APACHEⅡ score and B line score were not statistically significant (F value was 0.304, 0.404, 1.172, 1.403, andP value was 0.875, 0.805, 0.327, and 0.591, respectively). Pearson correlation analysis showed that there was a significant negative correlation between B line score and PaO2/FiO2 before and after treatment (r value was -0.579, -0.522, and -0.386, allP < 0.01), indicating that the more the B line, the more severe the pulmonary edema, and the worse the oxygenation; with the decrease in B line after treatment, the pulmonary edema was gradually alleviated, and oxygenation was gradually improved. There was a significant positive correlation between B line score and APACHEⅡ score before and 24 hours after treatment (r value was 0.484 and 0.536, bothP < 0.01), indicating that the more the B line, the more severe the patient; with the decrease in B line after treatment, the patient's condition improved after treatment. There was only a weak correlation between B line score and PAP at 24 hours after treatment (r = 0.317,P = 0.014), indicating that PAP was not a sensitive indicator in the degree of pulmonary edema in patients.Conclusions The more the B line in patients with severe high altitude pulmonary edema,the more severe of the pulmonary edema, and the more severe of the patient. There was no significant correlation between the B line score and PAP. Pulmonary ultrasonography can still be used not only in the plain and low elevation areas, but in the high altitude areas, as a reliable method to evaluate the severity of pulmonary edema.