Objective:To discuss the effect of exosome(Exo)microRNA-761(miR-761)on the epithelial-mesenchymal transition(EMT)process of the osteosarcoma(OS)cells by regulating tumor-associated macrophage(TAM)polarization,and to clarify its related mechanism.Methods:The miR-761 plasmid and negative control(miR-NC)plasmid were transfected into the HEK293 cells,and the non-transfected cells were regarded as control group.The transfection efficiency was detected using real-time fluorescence quantitative PCR(RT-qPCR)method.The Exo containing miR-761 was isolated,and the morphology of Exo was observed by transmission electron microscope.The concentration and size distribution of Exo samples were detected by nanoparticle analyzer,and the expression of Exo surface marker protein was detected by Western blotting method.The human monocyte leukemia THP-1 cells were stimulated with phorbol 12-myristate 13-acetate(PMA)to become the M0 macrophages,which were then treated with Exo containing miR-761 and co-cultured with the OS MG63 cells to establish the co-culture system.The experiment was divided into M0 group,TAM group,miR-761 NC group,and miR-761 Exo group.The M0 macrophages were collected from various groups,and the positive rates of M1 macrophage marker CD86 and M2 macrophage marker CD206 in various groups were detected by flow cytometry;the protein expression levels of M1 macrophage secreted factors interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)and M2 macrophage secreted factors interleukin-10(IL-10)and transforming growth factor-β1(TGF-β1)in various groups were detected by Western blotting method.The M0 macrophages were treated with Exo containing miR-761 and co-cultured with MG63 cells to establish the co-culture system.The experiment was divided into control group,TAM group,miR-NC Exo+TAM group,and miR-761 Exo+TAM group.The MG63 cells in various groups were collected,and the fluorescence intensities of E-cadherin and Vimentin in the MG63 cells in various groups were observed by immunofluorescence staining;the expression levels of E-cadherin,Vimentin,and EMT regulation-related transcription factors Twist1,Snail,and Slug proteins in the cells in various groups were detected by Western blotting method;the numbers of invasion and migration cells in various groups were detected by Transwell chamber assay.Results:The HEK293 cells containing miR-761 were successfully obtained by transfection experiments,and the Exo was isolated.Compared with M0 group,the positive rate of CD86 of the macrophages in TAM group was decreased(P＜0.05),while the positive rate of CD206 was increased(P＜0.05),the expression levels of IL-1β and TNF-α proteins were decreased(P＜0.05),while the expression levels of IL-10 and TGF-β1 proteins were increased(P＜0.05).Compared with TAM group,the positive rate of CD86 of the macrophages in miR-761 Exo group was increased(P＜0.05),while the positive rate of CD206 was decreased(P＜0.05),the expression levels of IL-1β and TNF-α proteins were increased(P＜0.05),while the expression levels of IL-10 and TGF-β1 proteins were decreased(P＜0.05).Compared with control group,the fluorescence intensity of E-cadherin in the MG63 cells in TAM group was decreased,while the fluorescence intensity of Vimentin was increased,the expression level of E-cadherin protein was decreased(P＜0.05),while the expression levels of Vimentin,Twist1,Snail,and Slug proteins were increased(P＜0.05),and the numbers of invasion and migration cells were increased(P＜0.05).Compared with TAM group,the fluorescence intensity of E-cadherin in the MG63 cells in miR-761 Exo+TAM group was increased,while the fluorescence intensity of Vimentin was decreased,the expression level of E-cadherin protein was increased(P＜0.05),while the expression levels of Vimentin,Twist1,Snail,and Slug proteins were decreased(P＜0.05),and the numbers of invasion and migration cells were decreased(P＜0.05).Conclusion:The exo-delivered miR-761 can inhibit the EMT process of the OS cells,thereby inhibiting the cell migration and cell invasion;its mechanism may be related to regulating TAM polarization.

Objective:To evaluate the effects of thoracic paravertebral block (TPVB) on graft patency in off-pump coronary artery bypass grafting.Methods:Fifty American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients, aged 60-75 yr, weighing 50-80 kg, undergoing elective off-pump coronary artery bypass grafting under combined intravenous-inhalational anesthesia, were divided into 2 groups using a random number table method: control group (group C, n=30) and TPVB group (group T, n=20). In group T, TPVB was performed at T 4, 5 interspace under the guidance of ultrasound, a test dose of 1% lidocaine 5 ml was injected on both sides, and 2 min later 0.375% ropivacaine 15 ml was injected.According to the anatomy of coronary artery, the graft was divided into 4 parts: left internal thoracic artery-anterior descending branch (arterial graft), the middle branch or the first diagonal branch, blunt marginal branche, and right posterior descending branch or left ventricular posterior branch.The blood flow was measured and pulsatility index was calculated after graft transplantation.Central venous pressure, mean pulmonary artery pressure, cardiac output, systemic resistance index and pulmonary resistance index were recorded immediately after placement of floating catheter (T 1), immediately after sawing sternum (T 2), immediately after anastomosis of all grafts (T 3) and 5 min before leaving the room (T 4). The intraoperative cardiovascular adverse events and consumption of sufentanil were recorded. Results:Compared with group G, central venous pressure, systemic resistance index and pulmonary resistance index were significantly decreased, cardiac output were increased, the incidence of intraoperative tachycardia was decreased, the incidence of hypotension was increased, the consumption of sufentanil was reduced, the flow of left internal thoracic artery-anterior descending branch was increased, and the pulsatility index was decreased at T 3 and T 4 in group T ( P<0.05). Conclusion:TPVB can improve the patency of left internal thoracic artery-anterior descending branch in off-pump coronary artery bypass grafting.

Objective:To summarize the efficacy of general anesthesia for percutaneous pulmonary valve implantation (PPVI).Methods:The clinical data of 6 patients underwent PPVI under general anesthesia in Children′s Heart Center of Henan Provincial People′s Hospital from December 2017 to January 2020 were retrospectively analyzed.Systolic blood pressure, diastolic blood pressure, heart rate, central venous pressure, SpO 2 and regional cerebral oxygen saturation were recorded before anesthesia induction (T 1), after anesthesia induction (T 2), before beginning of surgery (T 3), before pulmonary valve implantation (T 4), during pulmonary valve implantation (T 5), immediately after pulmonary valve implantation (T 6) and when the patients left the operating room (T 7). Right ventricular systolic pressure, diastolic pressure, pulmonary artery systolic pressure and diastolic pressure were recorded at T 4 and T 6.The development of related complications during operation and the cardiac, liver and kidney functions before and after operation were recorded.The postoperative extubation time, intensive care unit stay time and hospital stay time were recorded. Results:Six patients (3 males, 3 females), aged (16±4) yr, weighing (41±12) kg, were analyzed.Compared with the value at T 1-4 and T 6, 7, systolic blood pressure, diastolic blood pressure, heart rate, regional cerebral oxygen saturation and SpO 2 were significantly decreased at T 5 ( P<0.05). Compared with the value at T 1-5, central venous pressure was significantly decreased at T 6, 7 ( P<0.05). Compared with the value at T 4, right ventricular diastolic pressure was significantly decreased, and pulmonary artery diastolic pressure was increased at T 6 ( P<0.05). No anesthesia- and surgery-related serious complications occurred among the patients.One patient was transferred to the ward after extubation in the operating room, and 5 patients were transferred to the intensive care unit after operation.All 6 patients were discharged successfully and entered the follow-up stage. Conclusion:General anesthesia provides better efficacy when used for PPVI, and hemodynamic monitoring of pulmonary circulation and systemic circulation should be strengthened during pulmonary valve implantation to maintain circulation stable.

Objective: To identify the risk factors for postoperative hyperlactatemia in the patients with type A aortic dissection. Methods: Medical records of patients with type A aortic dissection who underwent cardiovascular surgery from January 2012 to October 2017 were retrospectively collected.The patients were divided into hyperlactatemia group and non-hyperlactatemia group according to the occurrence of hyperlactatemia (blood lactic acid ≥6 mmol/L) at 8 h after surgery.The variables of which P values were less than 0.05 in univariate analysis would enter the logistic regression analysis to stratify the risk factors for postoperative hyperlactatemia in this type of patients. Results: A total of 295 patients were included, of which 80 cases developed postoperative hyperlactatemia, and the incidence was 27.1%.Logistic regression analysis showed that preoperative acute pericardial tamponade, intraoperative deep hypothermic circulatory arrest time>35 min) and massive transfusion of blood (>1 000 ml) within 8 h after operation were independent risk factors for postoperative hyperlactatemia in the patients with type A aortic dissection. Conclusion Preoperative acute pericardial tamponade, intraoperative deep hypothermic circulatory arrest time>35 min and massive transfusion of blood (>1 000 ml) within 8 h after operation are independent risk factors for postoperative hyperlactatemia in the patients with type A aortic dissection.

Objective To prepare the biodegradable polylactic acid glycolic acid (PLGA) copolymer.encapsulated vascular endothelial growth factor(VEGF) loaded fluorescent controlled release sub-microspheres,to understand the efficiency of microspheres loading and releasing VEGF and to observe in vitro microspheres degradation.Methods VEGF-loading PLGA sub-microspheres were prepared by the two-phase solvent evaporation method,the in vitro degradation of fluorescent microspheres was observed by the laser confocal scanning electron microscopy.The drug loading efficiency and drug release curve were observed by ELISA.Results The VEGF loading PLGA fluorescent microspheres were successfully prepared by using the two-phase solvent evaporation method.The microspheres morphology was normal by using the scanning electron microscope and laser confocal microscope.The particle size was 0.5-1.0 μm with the laser particle size analyzer.The distribution was homogeneous.The VEGF loading rate and encapsulation rate detected by the quantitative ELISA were 3.91％ and 51.42 ％ respectively.The fluorescent microscope observed their slow degradation.The VEGF gentle release was detected by the quantitative ELISA,which showing linear zero order release trend.Conclusion The drug loading efficiency of VEGF-loading PLGA microspheres with 0.5-1.0 μm diameter is higher with linear zero order release,which can be directly observed by fluorescent light.

Objective To evaluate the effect of ulinastatin (UT1) on the expression of aquaporin 1 (AQP1) and AQP5 in rats with acute lung injury induced by cardiopulmonary bypass (CPB).Methods Forty-eight clean-grade healthy adult male Sprague-Dawley rats,weighing 200-250 g,were divided into 3 groups (n=16 each) using a random number table method:sham operation group (Sham group),CPB group and UTI group.UTI 200 000 U/kg was injected intravenously at 10 min prior to CPB in UTI group.The model of CPB was established in CPB and UTI groups.The equal volume of normal saline was intravenously injected at 10 min prior to puncture or at 10 min prior to CPB in Sham and CPB groups.Rats were sacrificed,and lung tissues were excised for determination of weight to dry weight ratio (W/D ratio),expression of AQP1 and AQP5 (by immunohistochemistry),expression of AQP1 and AQP5 protein and mRNA (by real-time polymerase chain reaction or Western blot) and for examination of morphological structure (with a light microscope) and ultrastructure of lung tissues (with an electron microscope).Injured alveolar rate (IAR) and rates of AQP1 and AQP5 positive cells were calculated.Results Compared with Sham group,W/D ratio and IAR were significantly increased,rates of AQP1 and AQP5 positive ceils were decreased,and the expression of AQP1 and AQP5 protein and mRNA was down-regulated in CPB and UTI groups (P＜0.05).Compared with CPB group,W/D ratio and IAR were significantly decreased,rates of AQP1 and AQP5 positive cells were increased,and the expression of AQP1 and AQP5 protein and mRNA was up-regulated in UTI group (P＜0.05).The injury to morphological structure and ultrastructure was significantly attenuated in UTI group when compared with CPB group.Conclusion The mechanism by which UTI pretreatment reduces CPB-induced acute lung injury is related to up-regulating the expression of AQP1 and AQP5 in rats.

Objective To analyze the independent risk factors and complications for perioperative hyperbilirubinemia in Stanford type A aortic dissection undergoing operation and investigate the management strategy of perioperative hyperbilirubi-nemia. Methods Between January 2013 and January 2018 from the department of great vessel surgery of heart centre of,290 cases of patients with Stanford type A aortic dissection undergoing operation were collected consecutively,male 210 cases,fe-male 80 cases. The related data and perioperative peak hyperbilirubinemia were recorded. According to the perioperative peak hyperbilirubinemia,patients were divided into 2 groups:≥51. 3 μmol/ L group and < 51. 3 μmol/ L group. Univariate and lo-gistic regression analysis were used to identify the independent risk factors. The perioperative complications were also recorded. Results Preoperative total bilirubin ≥ 17. 1 μmol/ L(OR = 2. 105,95％ CI: 1. 153 - 3. 125,P = 0. 016),cardiopulmonary bypass time > 3. 5 h(OR = 1. 103,95％ CI: 1. 316 - 6. 151,P = 0. 031),a large number of hemolysis(OR = 1. 503,95％CI: 1. 506 - 6. 651,P = 0. 029),the input amount of 24 h allogeneic red blood cell > 2000 ml(OR = 1. 381,95％ CI:0. 956 - 2. 552,P = 0. 036)were the independent risk factors for perioperative hyperbilirubinemia. The incidence rate of post-operative acute hepatic failure(2. 5％ vs. 0,P = 0. 021)and artificial liver therapy(2. 5％ vs. 0,P = 0. 021)in≥51. 3μmol/ L group were significantly increased. The incidence rate of postoperative acute lung injury(37. 5％ vs. 25. 2％,P =0. 039)and acute kidney injury(38. 7％ vs. 19. 5％,P = 0. 035)in 51. 3 μmol/ L group were also significantly increased. The duration of mechanical ventilation[(4. 1 ± 1. 6)days vs. (2. 8 ± 1. 3)days,P < 0. 05]and ICU stay time[(5. 1 ± 2. 3)days vs. (3. 9 ± 1. 8)days,P = 0. 035]and hospitalization time[( 19. 3 ± 3. 1)days vs. ( 17. 3 ± 2. 5)days,P = 0. 035]were sig-nificantly prolonged. Temporary nerve dysfunction(52. 5％ vs. 32. 6％,P = 0. 002)and in-hospital mortality( 17. 5％ vs. 8. 1％,P = 0. 037)were significantly increased. Conclusion Preoperative total bilirubin ≥ 17. 1 μmol/ L,cardiopulmonary bypass time > 3. 5 h,a large number of hemolysis,the input amount of 24 h allogeneic red blood cell > 2000 ml were the in-dependent risk factors for perioperative hyperbilirubinemia in Stanford type A aortic dissection. The perioperative complications in≥51. 3 μmol/ L group were significantly increased. Therefore,more attention should be paid to the independent risk factors for perioperative hyperbilirubinemia in Stanford type A aortic dissection,hyperbilirubinemia and its clearance should be moni-tored more actively and dynamically,the cause should be found more precisely,the treatment be more comprehensive to achieve to control the level of bilirubinemia and improve the prognosis.

Objective To evaluate the effect of ulinastatin pretreatment on endoplasmic reticulum stress during myocardial injury in the patients undergoing mitral valve replacement (MVR) with cardiopulmonary bypass (CPB).Methods One hundred patients of both sexes,aged 35-64 yr,weighing 40-80 kg,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ (New York Heart Association Ⅱ or Ⅲ),scheduled for elective MVR with CPB,were divided into ulinastatin pretreatment group (UP group,n=50) and normal saline control group (NS group,n =50) using a random number table.Ulinastatin 0.5× 104 U/kg was intravenously infused over 1 h before skin incision,and administration was repeated every 4 h until the end of operation in group UP,while the equal volume of normal saline was given instead in group NS.Immediately after opening the right atrium (T0),at 30 min after aortic clamping (T1) and while suturing the right atrium (T2),blood samples were collected from the radial artery for measurement of the concentrations of plasma creatine kinase-MB and cardiac troponin T by enzyme-linked immunosorbent assay.Right auricle specimens were obtained after blood sampling at each time point for determination of the expression of glucose-regulated protein 78,CCAAT/enhancer-binding protein homologous protein and c-Jun N-terminal kinase protein and mRNA (by real-time polymerase chain reaction and Western blot,respectively) and apoptosis in cardiomyocytes (by TUNEL).The apoptosis rate was calculated.Results Compared with group NS,the plasma concentrations of creatine kinase-MB and cardiac troponin T at T1 and T2 and apoptosis rate at T2 were significantly decreased,and the expression of glucose-regulated protein 78,CCAAT/enhancer-binding protein homologous protein and c-Jun N-terminal kinase protein and mRNA was down-regulated at T1 and T2 in group UP (P＜0.05).Conclusion The mechanism by which ulinastatin pretreatment inhibits apoptosis in cardiomyocytes and attenuates myocardial injury is related to decrease in endoplasmic reticulum stress in the patients undergoing MVR with CPB.

Objective To analyze of the efficacy of minimally invasive surgery combined with medication in the treatment of ureteral calculi.Methods From June 2013 to June 2015,124 patients with ureteral calculi which confirmed by X-ray in our hospital were collected as subjects.According to the treatment method,they were divided into control group and observation group,patients in the control group were treated with simple medication,while the observation group were treated with minimally invasive surgery combined with drug therapy.The effects of treatment and the levels of MCP-1,TFF1 and HMGB1 in urine between two groups were compared.Results The stone expulsion rate and the net rate of the observation group was higher than those of the control group,the differences were significant(P<0.05).The stone expulsion time of observation group was shorter than that of the control group,the formation rate of the observation group was lower than that of the control group,the differences were significant(P<0.05).Adverse reactions occurred in 11 cases of the observation group,in 29 cases of the control group,the incidence of adverse reactions in the observation group was lower than that in the control group,the difference was significant(P<0.05).After the treatment,17 cases of adverse drug reactions occurred in the observation group,which was more than 5 cases of adverse drug reactions in the control group,the difference was significant(P<0.05).After treatment,the levels of MCP-1 and HMGB1 in urine of two groups were decreased compared with those before treatment,the level of TFF1 was increased,the differences were significant(P<0.05);while the levels of MCP-1 and HMGB1 of the observation group after treatment were lower than those of the control group,the level of TFF1 was higher than that of the control group,the difference was significant(P<0.05).The effective rate of the observation group was 88.71％,which higher than 69.35％ of the control group,the difference was significant(P<0.05).Conclusion On the basis of medication,combined with extracorporeal shock wave lithotripsy for ureteral calculi can not only improve the therapeutic effect,but also reduce the risk of adverse reactions.

BACKGROUND:There are many complications of limb salvage surgery in patients with osteosarcoma of the middle tibia, and the limb salvage surgery is one of the current difficulties in clinical treatment. OBJECTIVE:To evaluate the clinical efficacy of reconstruction with massive al ograft bone for osteosarcoma of the middle tibia by retrospectively reviewing relevant cases. METHODS:Seven patients with osteosarcoma of the middle tibia were treated. And we analyzed their clinical data retrospectively. Al patients completed the formal preoperative adjuvant chemotherapy and we confirmed that there was no distant metastasis before surgery. Al patients received large al ogeneic bone transplantation and internal fixation, and the gastrocnemius muscle flap coveraged graft bone in surgery. The average length of al ogeneic bone was 12.5 cm. Five patients received postoperative adjuvant chemotherapy completely, and two patients received partly. RESULTS AND CONCLUSION:The fol ow-up period was 18-36 months. One patient had local tumor recurrence at 1 year after transplantation, and died of lung metastases after amputation. One patient survived after resection of lung metastases that occurred at 1.5 years after transplantation. One patient died of lung metastases at 2 years after transplantation. The rest four patients were tumor-free. The mean Musculoskeletal Tumor Society (MSTS) score was 26.5, the mean International Society of Limb Salvage (ISOLS) graft score was 31. Among four underage patients, one had leg length deformities, with limb shortening 2 cm. There were no postoperative infections and pathological fractures. Using large al ogeneic bone for the repair of bone defects after tumor surgery of the middle tibia can have a good clinical efficacy under the premise of strict indications. Using gastrocnemius muscle flap to cover the bone graft during surgery is an effective measure to reduce postoperative complications.