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 study the risk factors of surgical therapy in neonates with necrotizing enterocolitis (NEC).Methods:From January 2016 to July 2020, neonates with a confirmed diagnosis of NEC (Bell's Stage Ⅱ and above) admitted to our hospital were retrospectively enrolled. They were assigned into surgical group and conservative group according to whether surgeries were performed. The conditions during perinatal period, clinical characteristics and laboratory examinations at the onset of NEC were compared between the two groups. Multivariate Logistic regression analysis was used to determine the risk factors of surgical therapy.Results:A total of 177 neonates with NEC were identified, including 62 cases (35.0%) in the surgical group and 115 cases (65.0%) in the conservative group. Multivariate Logistic regression analysis showed that male gender ( OR=3.178,95% CI 1.457~6.929, P=0.004), comorbidity with shock ( OR=3.434, 95% CI 1.112~10.607, P=0.032), mechanical ventilation>7 d before NEC onset ( OR=3.663, 95% CI 1.098~12.223, P=0.035) and lymphocytes <2.0×10 9/L ( OR=4.121, 95% CI 1.801~9.430, P=0.001) at the onset of NEC were independent risk factors for surgical therapy. Conclusions:Male gender, comorbidity with shock, mechanical ventilation >7 d before NEC and lymphocytopenia at the onset are independent risk factors for surgical therapy in neonates with NEC (Stage Ⅱ and above).

Objective:To investigate the early and middle term clinical efficacies of 3D-printed metal prostheses in the reconstruction of bone defects after osteotomy in malignant bone tumors.Methods:A total of 34 patients with malignant bone tumors of lower extremity femur and tibia who underwent 3D printing individualized metal prosthesis replacement surgery in the Department of Bone and Soft Tissue of Affiliated Cancer Hospital of Zhengzhou University from March 2019 to March 2022 were retrospectively analyzed. There were 23 males and 11 females, with an average age of 19.1±15.2 years (range, 7-80 years). There were 22 children and adolescents younger than 18 years old. There were 3 cases in the proximal femur, 15 cases in the middle and distal femur, 10 cases in the proximal tibia and 6 cases in the distal tibia. According to the final pathological diagnosis, 24 cases of osteosarcoma, 6 cases of Ewing's sarcoma, 2 cases of undifferentiated sarcoma, 1 case of osteosarcoma, and 1 case of malignant giant cell tumor of bone were enrolled in this study. Postoperative complications, wound healing, periprosthetic fracture and aseptic loosening, tumor outcome (evaluated by tumor control evaluation criteria), and length difference of lower limbs were recorded. Response evaluation criteria in solid tumor (RECIST) was used to evaluate tumor outcomes. Prosthetic-bone interface healing was evaluated postoperatively, and the function was evaluated based on Musculoskeletal Oncology Society (MSTS) 93.Results:The length of lesions was 70-240 mm in 34 patients, with an average of 125.5±35.4 mm. The length of osteotomy was 80-275 mm, with an average of 160.2±33.9 mm. No tumor was found on the osteotomy surface. The customized prosthesis was firmly installed and closely matched with the side of the preserved articular surface. There were 2 patients with local incision fat liquefaction and 4 patients with superficial wound infection, which healed after debridement and antibiotic treatment. One distal tibia osteosarcoma case developed severe periprosthetic infection 2 months after surgery, resulting in prosthesis implantation failure, limb movement pain and poor ankle function. After removal of the prosthesis, infection control and osteogenesis with the Ilizarov technique, the infection was completely controlled and local osteogenesis was possible. The remaining 33 patients had a good prosthetic-bone interface union. One case was found to have localized bone resorption on the contact surface of the prosthesis 7 months after operation, but the metal prosthesis and screws were not loose. The incisions healed well in other patients, without infection, prosthesis loosening, fracture or other complications. All patients survived and were followed up for 13.8±5.6 months (range, 7-27 months). During the follow-up, there was no recurrence of tumor at the osteotomy end in all patients, but 5 patients developed lung metastasis. At the end of the last follow-up, all patients survived. Among them, 16 patients had unequal length of lower limbs, including 10 cases within 2 cm, 3 cases between 2-5 cm, and 3 cases over 5 cm. With the exception of one patient whose prosthesis was removed due to infection, the MSTS 93 of the other patients was 24.9±2.2 (range, 19-28), and were rated as excellent in 26 cases and good in 7 cases. According to the RECIST evaluation criteria, 26 of 34 patients had complete response, 5 had disease progression, and 3 had stable disease.Conclusion:3D printed metal prosthesis is one of the effective methods for the treatment of bone defects after resection of malignant bone tumors in lower limbs, which is safe, reliable and has satisfactory early curative effect.

Objective:To assess the effectiveness of percutaneous acetabuloplasty (PA) in combination with radiotherapy for the treatment of acetabular metastases.Methods:A retrospective analysis of medical records from 43 patients with acetabular metastases admitted between May 2017 and May 2022 were performed, with 24 cases meeting inclusion criteria. The study cohort consisted of 9 males and 15 females, with an average age of 56.0 years (range: 40-85 years). There were 12 cases on the left side and 12 cases on the right side. The primary cancer types were breast cancer (8 cases, 33%), lung cancer (7 cases, 29%), prostate cancer (4 cases, 17%), bowel cancer (2 cases, 8%), cervical cancer (1 case, 4%), kidney cancer (1 case, 4%), and liver cancer (1 case, 4%). All patients had multiple bone metastases, with 16 cases (67%) also presenting with metastases in other organs. Among the bone metastases, 19 cases (79%) were osteolytic lesions, and 5 cases (21%) were mixed lesions. Lesion distribution included 11 cases in the acetabulum, 2 cases in the acetabulum and anterior column, 8 cases in the acetabulum and posterior column, and 3 cases in the acetabulum, anterior column, and posterior column. Lesion sizes ranged from 2.0 cm×1.5 cm×2.0 cm to 4.5 cm×4.0 cm×11.0 cm. Cortical defects were observed in 11 cases (46%), and soft tissue masses were present around the acetabular metastasis in 8 cases (33%). PA was performed under local or general anesthesia, followed by local radiotherapy within 1 week after surgery (external radiotherapy, 30 Gy, 10 d). Various clinical parameters, including primary lesion location, time of tumor diagnosis, time of bone metastasis diagnosis, number and nature of bone metastases, distribution area, lesion size, presence of cortical defects and soft tissue masses, presence of other organ metastases, surgical site, operation duration, filling effect, complications, visual analog scale (VAS) scores, walking scores, Eastern Cooperative Oncology Group (ECOG) scores, and long-term complications, were recorded and compared before surgery, after surgery, after radiotherapy, at 1, 3, and 6 months post-surgery, and during the last follow-up. The median follow-up period was 18 months.Results:Among the 24 patients, the procedure was successfully completed in 23 cases and failed in 1 case due to puncture needle-related complications. At the last follow-up, 92% (22/24) of patients showed no local symptom progression. VAS score, walking score, and ECOG score improved from 7.2±1.1, 1.4±1.4, 2.5±0.7 before surgery to 2.6±1.9, 2.5±1.4, 2.0±0.8 at 48 hours post-operation. There were no significant differences in scores between 48 hours post-surgery and 48 hours post-radiotherapy.Conclusion:PA can rapidly restore acetabular stability, alleviate pain, and enhance the quality of life for patients. In cases of poor response to PA, radiotherapy may not be effective, but it can effectively control local symptom progression. The combination of these two interventions can yield satisfactory clinical outcomes for patients with acetabular metastases.

【Objective】 To study the annual financial expenditure in blood stations with different scales, and to establish the regression equation between blood collection units and total expenditure. 【Methods】 The annual total expenditure, the per capita cost of serving population, as well as the collection units of whole blood and apheresis platelet of 24 blood stations were collected. The financial expenditure required for collecting 10 000U blood was calculated.The statistical analysis was carried out with SPSS statistical software. 【Results】 From 2017 to 2020, the total annual financial expenditure of 24 blood stations showed an upward trend. The total expenditure among blood stations was different. The per capita cost of servicing population in the areas where the 24 blood stations were located had been increasing year by year. The 24 blood stations were divided into two grades according to the blood collection volume as 50 000 U, and the relationship equation between the blood collection volume and the annual total expenditure had been established. After testing, each equation was effective(P<0.05); There was no difference in the financial expenditure required for collecting 10 000U blood among blood stations with different scales. 【Conclusion】 From 2017 to 2020, the blood stations with an annual collection volume more than 50 000 U demonstrated a higher financial expenditure and the per capita cost of serving population than those <50 000 U. The blood collection volume of blood stations is significantly correlated with the annual total expenditure and the per capita cost of serving population.

Objective:To explore the possible clinical benefits of CT/MRI image fusion and computer-assisted simulation techniques in guiding type Ⅲ and Ⅳ primary pelvic bone tumor surgeries.Methods:The clinic data of primary bone sarcomas patients treated at Department of Bone and Soft Tissue,Zhenghzhou University Affiliated Cancer Hospital from January 2019 to December 2021 were retrospectively analyzed. Based on whether the CT and MRI image fusion technique was utilized for tumor evaluation and surgical planning,the patients were divided into image fusion group ( n=21) or control group ( n=27). There were 7 male and 14 female patients included in the image fusion group, with the age of (37.0±10.4) years(range: 18 to 67 years). In the control group, there were 10 males and 17 females with the age of (39.7±15.2) years (range: 16 to 65 years). Both groups included osteosarcoma,chondrosarcoma and undifferentiated polymorphic sarcoma as the pathological diagnosis. Clinical information such as gender,age,pathological diagnosis,location of disease,and metastasis at diagnosis were collected. Surgical related information such as duration of surgery,blood loss,surgical margin,and wound complications were also obtained. Periodical follow-ups every 3 months were performed for all patients to monitor the status of local recurrence,distant metastasis,and survival information. Independent t test and χ2 test were used for data comparison between groups. Results:Significant reduced duration of surgery was observed in the image fusion group in comparison with control group both in type Ⅲ and Ⅳ surgeries ((144.0±31.6)min vs. (248.2±56) min, t=-8.084, P<0.01); (173.0±42.0)min vs. (306.1±62.0)min, t=-4.518, P<0.01). Blood loss was significantly reduced in the image fusion group compared with the control group ((484.8±226.3)ml vs. (836.1±359.8)ml, t=-4.130, P<0.01). In addition, significant lower ratio of R1 margin and recurrence rates of type Ⅲ and Ⅳ surgeries were found in the image fusion group comparing with the control group (4.8%(1/21) vs. 22.2%(6/27), χ2=4.214, P=0.040; 4.8%(1/21) vs. 22.2%(6/27), χ2=4.214, P=0.040).In the image fusion group, there were 3 cases of incision infection, 1 of which underwent secondary debridement.And in thecontrol group there were 7 cases of incision infection, 3 of which underwent secondary debridement. There was no significant difference in the incidence of complications between the two groups (14.2%(3/21) vs. 25.9%(7/27), χ2=0.645, P=0.422). Up to the last follow-up, 1 patient died in the image fusion group and 2 patients died in the control group, the difference was not statistically significant (χ2=1.885, P=0.220). Conclusion:Compared with the traditional operation,the image fusion technique can significantly reduce the duration of surgery,blood loss and lower the recurrence rate by achieving better surgical margins.

Objective:To explore the possible clinical benefits of CT/MRI image fusion and computer-assisted simulation techniques in guiding type Ⅲ and Ⅳ primary pelvic bone tumor surgeries.Methods:The clinic data of primary bone sarcomas patients treated at Department of Bone and Soft Tissue,Zhenghzhou University Affiliated Cancer Hospital from January 2019 to December 2021 were retrospectively analyzed. Based on whether the CT and MRI image fusion technique was utilized for tumor evaluation and surgical planning,the patients were divided into image fusion group ( n=21) or control group ( n=27). There were 7 male and 14 female patients included in the image fusion group, with the age of (37.0±10.4) years(range: 18 to 67 years). In the control group, there were 10 males and 17 females with the age of (39.7±15.2) years (range: 16 to 65 years). Both groups included osteosarcoma,chondrosarcoma and undifferentiated polymorphic sarcoma as the pathological diagnosis. Clinical information such as gender,age,pathological diagnosis,location of disease,and metastasis at diagnosis were collected. Surgical related information such as duration of surgery,blood loss,surgical margin,and wound complications were also obtained. Periodical follow-ups every 3 months were performed for all patients to monitor the status of local recurrence,distant metastasis,and survival information. Independent t test and χ2 test were used for data comparison between groups. Results:Significant reduced duration of surgery was observed in the image fusion group in comparison with control group both in type Ⅲ and Ⅳ surgeries ((144.0±31.6)min vs. (248.2±56) min, t=-8.084, P<0.01); (173.0±42.0)min vs. (306.1±62.0)min, t=-4.518, P<0.01). Blood loss was significantly reduced in the image fusion group compared with the control group ((484.8±226.3)ml vs. (836.1±359.8)ml, t=-4.130, P<0.01). In addition, significant lower ratio of R1 margin and recurrence rates of type Ⅲ and Ⅳ surgeries were found in the image fusion group comparing with the control group (4.8%(1/21) vs. 22.2%(6/27), χ2=4.214, P=0.040; 4.8%(1/21) vs. 22.2%(6/27), χ2=4.214, P=0.040).In the image fusion group, there were 3 cases of incision infection, 1 of which underwent secondary debridement.And in thecontrol group there were 7 cases of incision infection, 3 of which underwent secondary debridement. There was no significant difference in the incidence of complications between the two groups (14.2%(3/21) vs. 25.9%(7/27), χ2=0.645, P=0.422). Up to the last follow-up, 1 patient died in the image fusion group and 2 patients died in the control group, the difference was not statistically significant (χ2=1.885, P=0.220). Conclusion:Compared with the traditional operation,the image fusion technique can significantly reduce the duration of surgery,blood loss and lower the recurrence rate by achieving better surgical margins.

Objective:To retrospectively analyze the clinical effects and complications of alcohol inactivation, irradiation inactivation, and liquid nitrogen inactivation in the treatment of femur osteosarcoma in children, in an attempt to provide a theoretical basis for clinical selection of in vitro inactivation methods of tumor bone segment. Methods:The clinical data of 93 children with femur osteosarcoma admitted to the Department of Bone and Soft Tissue, the Affi-liate Cancer Hospital of Zhengzhou University from January 2008 to December 2017 were retrospectively analyzed, and 40 children, including 21 males and 19 females, aged 8-18 (13.65±2.87) years, who were treated with in vitro inactivation and replantation of autogenous tumor bone segment, were screened.Among these children, there was alcohol inactivation in 15 cases, irradiation inactivation in 12 cases, and liquid nitrogen inactivation in 13 cases.A comparison was drawn on these 3 inactivation methods with respect to bone healing time, bone healing rate, tumor recurrence rate, infection rate, fracture or fixation failure rate, and revision rate. Results:All those 40 children were subject to valid medical followed-up, with the alcohol inactivation group for (102.60±16.55) months, the irradiation inactivation group for (59.33±6.39) months, and the liquid nitrogen inactivation for (36.85±6.49) months.The difference in follow-up time of 3 groups was statistically significant ( P<0.05). Compared with other 2 groups, the index of bone healing time, bone healing rate, infection rate and revision rate in the alcohol inactivation group were unfavorable, which showed a significant difference (all P<0.05); However, there was no significant difference in the recurrence rate, fracture rate or fixation failure rate compared with other 2 groups (all P>0.05); There was no significant difference in all above indexes between the irradiation group and the liquid nitrogen group (all P>0.05). Conclusions:Three in vitro inactivation methods for the treatment of tumor bone segment are safe and reliable.The alcohol inactivated bone has a long healing time and more complications.Both irradiation inactivation and liquid nitrogen inactivation are clinical options, but irradiation inactivation requires professional equipment, which may limit the clinical application.

Objective:To study the risk factors of necrotizing enterocolitis (NEC) after surgery for intestinal atresia.Method:From August 2013 to June 2020, children with intestinal atresia receiving surgery in our hospital were retrospectively reviewed. The patients were assigned into NEC group and non-NEC group according to the occurrence of postoperative NEC. Demographic data and clinical characteristics were summarized and the risk factors for postoperative NEC were analyzed using Logistic regression analysis method.Result:A total of 96 infants were enrolled and NEC occurred in 13 patients (13.5%) after surgery for intestinal atresia. Compared with the non-NEC group, the NEC group were diagnosed of intestinal atresia [4.0(1.5,6.0)d vs. 1.4(0,2.0)d, P<0.001] and received surgery [4.8(2.0,7.0)d vs. 3.1(1.0,4.0)d, P=0.034] at later ages. The incidences of complex intestinal atresia [76.9%(10/13) vs. 44.6%(37/83), P=0.030] and blood transfusion [46.2%(6/13) vs. 13.3%(11/83), P=0.007] in the NEC group were higher than the non-NEC group. Logistic regression analysis showed that the age of initial diagnosis of intestinal atresia ( OR=3.346, 95% CI 1.493~7.500, P=0.003), complex intestinal atresia ( OR=9.052, 95% CI 1.119~73.209, P=0.039) and blood transfusion ( OR=6.835, 95% CI 1.399~33.380, P=0.018) were independent risk factors for postoperative NEC. Conclusion:Patients with delayed diagnosis of intestinal atresia, complex intestinal atresia and blood transfusion within 48 hours after surgery should be monitored for the occurrence of postoperative NEC.

Objective To investigate the clinical effect of using pedicled flap to reconstruct skin and soft tis-sue defect after resection of sarcoma around extremities joint. Methods Retrospectively analysis 103 cases of sar-coma around extremities joint who was treated from June, 2012 to June, 2017 and used the pedicled fasciocutaneous flaps or pedicled musculocutaneous flaps to reconstruct the defect after sarcoma resection. There were 61 males and 42 females, with a mean age of 52.3 (14-82 ) years old. Among these patients, there were 32 initial diagnosis, 30 had been resected in other units, and 41 recurrent tumor. There were 13 patients that tumor around wrist:reconstructed by radial wrist upper branch fasciocutaneous flap, dorso-ulnar flap based on supracarpal perforator, and posterior in-terosseous artery flap. Eight patients with tumor around elbow: reconstructed by lateral arm flap and media arm flap. Eight patients with tumor around shoulder: reconstructed by latissimus dorsi musculocutaneous flap and lateral tho-racic flap. Fourteen patients with tumor around hip: rectus femoris musculocutaneous flap, tensor fascia lata musculo-cutaneous flap, and posterior thigh fasciocutaneous flap. Sixteen patients that tumor around knee:reconstructed by an-terolateral low leg flap, and sural calf fasciocutaneous flap. Forty-four patients that tumor around ankle: reconstructed by saphenous neuro-veno-fasciocutaneous flap, sural neuro-veno-fasciocutaneous flap, lateral supramalleolar flap, and medial plantar flap. The defect area after tumor resection were:3 cm ×4 cm-18 cm×25 cm. After surgery, observe the situation and fitness of flap, complication, joint mobility, and tumor recurrence and metastasis. Results Allflaps survived except 6 died or partial died, with a survival rate of 94.2％, of which dead flap were replaced by a new flap or covered by free skin graft. Infection occurred in 4 cases (2 in donor site and 2 in recipient site), were cured by dressing and antibiotic. Except for 12 cases dropped, all rest 91 cases were followed-up by 33 months on average (1-5 years), flaps' shape and texture were good, without swollen and fester, and fit well. Superficial sensibility of direct fasciocuta-neous flaps and musculocutaneous flaps were normal, protective sensation of reverse fascial flaps were recovered. The mobility of affected joint show no difference compared with the other side. No ulcer occurred on flaps of plantar and but-tocks, that helps normal sit position and gait. There were 17 recurrences and 32 metastasis occurred, 21 patients died so far. There was a statistical difference between recurrent cases with first shot(10.3％) second widely resection(11.5％) and recurrent resection(30.6％)(P<0.05). Conclusion Using pedicled flap to reconstruct skin defect after the resection of sarcoma around extremities joint, can get good clinical result, with simple and reliable operation. It is worth to popularize.