Objective:To investigate the clinical efficacy of liquid nitrogen cryopreservation and reimplantation versus allograft reconstruction in patients underwent resection of primary malignant bone tumors of the long bones of the extremities.Methods:A retrospective analysis was conducted on 144 patients who underwent resection of primary malignant bone tumors of the long bones of the extremities followed by either liquid nitrogen cryopreservation and reimplantation or massive allografts reconstruction at the Beijing Jishuitan Hospital affiliated with Capital Medical University from January 2012 to July 2023. The study included 82 males and 62 females, with an average age of 23.8±12.3 years (range, 6-64 years). Patients were divided into two groups based on the reconstruction method: the cryopreservation and reimplantation group (72 cases) and the allograft group (72 cases). The following outcomes were recorded during follow-up: local tumor recurrence, bone union, union time, graft survival, and reasons for graft removal. Graft-related complications were recorded using the modified Henderson classification system of the International Limb Salvage Association. Limb function was assessed at the last follow-up using the Musculoskeletal Tumor Society score (MSTS-93).Results:All patients completed surgery and were followed up for a mean of 60.2±32.1 months (range, 12-149 months). At the last follow-up, 24 patients were dead from the tumor, 16 patients survived with the tumor (2 cases of local recurrence and 14 cases of distant metastasis), and 104 patients survived without the tumor. The bone union rate and union time in cryopreservation and reimplantation group were 90% (65/72) and 9.6±4.8 months, respectively, which was significantly superior to those in allograft group [68% (49/72) and 15.9±6.7 months, P<0.05]. The 5-year overall graft survival rate was 86.8% [95% CI (80.1%, 95.7%)] in cryopreservation and reimplantation group, higher than 73.2% [95% CI(68.4%, 84.5%)] in allograft group significantly (χ 2=7.122, P=0.017). The rates of graft removal due to non-union and infection in the cryopreservation and reimplantation group were 0% (0/72) and 1.4% (1/72), respectively, which were significantly lower than those in the allograft group [5.6% (4/72) and 9.7% (7/72), P<0.05]. Overall, 48.6% (70/144) of patients experienced at least one graft-related complication, with a complication rate of 33.3% in cryopreservation and reimplantation group, lower than the 61.1% in allograft group significantly (χ 2=11.146, P<0.001). The complications with the highest incidence rate were nonunion (20.8%, 30/144), followed by structural failure (17.4%, 25/144), tumor progression (10.4%, 15/144), infection (10.4%, 15/144), and soft tissue failure (5.6%, 8/144). The incidence rates of the atrophic non-union and the structural failure of grafts were 9.7% (7/72) and 1.4% (1/72) respectively in the cryopreservation and reimplantation group, which were significantly lower compared to the allograft group [29.2% (21/72) and 13.9% (10/72), P<0.05]. At the last follow-up, the MSTS-93 score was 89.7%±8.3% in the cryopreservation and reimplantation group, and 87.6%±7.5% in the allograft group, with no statistically significant difference ( t=0.326, P=0.542). Conclusion:Compared with allograft reconstruction, autologous inactivated bone grafting demonstrated superior bone union efficiency and fewer complications, it may be considered for reconstruction in cases whose tumor bone is not severely osteolytic or pathologically fractured.

Objective:To investigate the clinical efficacy of liquid nitrogen cryopreservation and reimplantation versus allograft reconstruction in patients underwent resection of primary malignant bone tumors of the long bones of the extremities.Methods:A retrospective analysis was conducted on 144 patients who underwent resection of primary malignant bone tumors of the long bones of the extremities followed by either liquid nitrogen cryopreservation and reimplantation or massive allografts reconstruction at the Beijing Jishuitan Hospital affiliated with Capital Medical University from January 2012 to July 2023. The study included 82 males and 62 females, with an average age of 23.8±12.3 years (range, 6-64 years). Patients were divided into two groups based on the reconstruction method: the cryopreservation and reimplantation group (72 cases) and the allograft group (72 cases). The following outcomes were recorded during follow-up: local tumor recurrence, bone union, union time, graft survival, and reasons for graft removal. Graft-related complications were recorded using the modified Henderson classification system of the International Limb Salvage Association. Limb function was assessed at the last follow-up using the Musculoskeletal Tumor Society score (MSTS-93).Results:All patients completed surgery and were followed up for a mean of 60.2±32.1 months (range, 12-149 months). At the last follow-up, 24 patients were dead from the tumor, 16 patients survived with the tumor (2 cases of local recurrence and 14 cases of distant metastasis), and 104 patients survived without the tumor. The bone union rate and union time in cryopreservation and reimplantation group were 90% (65/72) and 9.6±4.8 months, respectively, which was significantly superior to those in allograft group [68% (49/72) and 15.9±6.7 months, P<0.05]. The 5-year overall graft survival rate was 86.8% [95% CI (80.1%, 95.7%)] in cryopreservation and reimplantation group, higher than 73.2% [95% CI(68.4%, 84.5%)] in allograft group significantly (χ 2=7.122, P=0.017). The rates of graft removal due to non-union and infection in the cryopreservation and reimplantation group were 0% (0/72) and 1.4% (1/72), respectively, which were significantly lower than those in the allograft group [5.6% (4/72) and 9.7% (7/72), P<0.05]. Overall, 48.6% (70/144) of patients experienced at least one graft-related complication, with a complication rate of 33.3% in cryopreservation and reimplantation group, lower than the 61.1% in allograft group significantly (χ 2=11.146, P<0.001). The complications with the highest incidence rate were nonunion (20.8%, 30/144), followed by structural failure (17.4%, 25/144), tumor progression (10.4%, 15/144), infection (10.4%, 15/144), and soft tissue failure (5.6%, 8/144). The incidence rates of the atrophic non-union and the structural failure of grafts were 9.7% (7/72) and 1.4% (1/72) respectively in the cryopreservation and reimplantation group, which were significantly lower compared to the allograft group [29.2% (21/72) and 13.9% (10/72), P<0.05]. At the last follow-up, the MSTS-93 score was 89.7%±8.3% in the cryopreservation and reimplantation group, and 87.6%±7.5% in the allograft group, with no statistically significant difference ( t=0.326, P=0.542). Conclusion:Compared with allograft reconstruction, autologous inactivated bone grafting demonstrated superior bone union efficiency and fewer complications, it may be considered for reconstruction in cases whose tumor bone is not severely osteolytic or pathologically fractured.

OBJECTIVE To investigate the mechanism by which salvianolic acid A (Sal A) reduces the inflammatory response and oxidative stress of BV2 cells injured by oxygen and glucose deprivation/reperfusion (OGD/R).METHODS An OGD/R injury model of BV2 cells was established with sugar free Earle solution containing Na2S2O410 mmol·L-1.Na2S2O4 sugar free Earle solution was added and cultured in an incubator (37 ℃,5％CO2) for 1.5 h (oxygen glucose deprivation) before a normal medium was used for 24 h (reperfusion).Then,the cells were divided into the cell control group,OGD/R group,OGD/R+Sal A 1,5 and 10 μmol·L-1 group,OGD/R+ML385 group,OGD/R+ML385+Sal A 1,5 and 10μmol·L-1 group and OGD/R+edaravone (Eda,50μmol·L-1) group.After twenty-four hours of culture,the cell survival rate was measured by CCK8 kit.The contents of Interleukin-1β (IL-1β),IL-6,tumor necrosis factor-α(TNF-α),IL-10,IL-4 and transforming growth factor-β(TGF-β) in the cell supernatant were detected by ELISA.Reactive oxygen species (ROS) in cells was detected using the chemical fluo-rescence method.The contents of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD),glutathione peroxidase (GSH-PX) and chloramphenicol acetyltransferase (CAT) in cells were determined with the colorimetric method.Protein expressions of Kelch like ECH-associated protein 1 (Keap1),nuclear factor erythroid-2 related factor 2 (Nrf2),Heme oxygenase-1 (HO-1),NADPH:quinone oxidoreductase-1 (NQO1) and p-nuclear factor kappa-B p65 (p-NF-κB p65) were detected by Western blotting.RESULTS ①Compared with the cell control group,the cell survival rate of the OGD/R group was significantly decreased (P＜0.01).Compared with the OGD/R group,the survival rates of OGD/R+Sal A 1,5 and 10μmol·L-1 groups were significantly increased (P＜0.05,P＜0.01).②Compared with the cell control group,the contents of IL-1β,IL-6 and TNF-α were significantly increased,the contents of IL-10,IL-4 and TGF-β were significantly decreased,the contents of ROS and MDA were significantly increased,and the activities of SOD,CAT and GSH-Px were significantly decreased in the OGD/R group (P＜0.01).Compared with the OGD/R group,the content of IL-6 was significantly decreased,the contents of IL-10,IL-4 and TGF-β were significantly increased,the contents of ROS and MDA were significantly decreased,and the activities of SOD,CAT and GSH-Px were significantly increased in OGD/R+Sal A 1,5 and 10μmol·L-1 and OGD/R+Eda groups (P＜0.05,P＜0.01).③Compared with the cell control group,the protein expression of p-NF-κB P65 in the OGD/R group was significantly increased (P＜0.01).Compared with the OGD/R group,the protein expressions of Keap1 and cytoplasmic Nrf2 were significantly decreased,the expressions of nuclear Nrf2,HO-1 and NQO1 proteins were significantly increased,and the expression of p-NF-κB p65 protein was significantly decreased in OGD/R+Sal A 5 and 10 μmol·L-1 and OGD/R+Eda groups (P＜0.05,P＜0.01).In OGD/R+ML385,OGD/R+ML385+Sal A 1,5 and 10μmol·L-1 groups,the protein expression of Keap1 was significantly increased,the protein expressions of cytoplasmic Nrf2,nuclear Nrf2,HO-1 and NQO1 protein were significantly decreased,and the protein expression of p-NF-κB P65 was significantly increased (P＜0.01).CONCLU-SION Sal A reduces the inflammatory response and oxidative stress of OGD/R injured BV2 cells possi-bly by activating the Keap1/Nrf2 pathway and inhibiting the NF-κB pathway.

OBJECTIVE To investigate the mechanism by which salvianolic acid A (Sal A) reduces the inflammatory response and oxidative stress of BV2 cells injured by oxygen and glucose deprivation/reperfusion (OGD/R).METHODS An OGD/R injury model of BV2 cells was established with sugar free Earle solution containing Na2S2O410 mmol·L-1.Na2S2O4 sugar free Earle solution was added and cultured in an incubator (37 ℃,5％CO2) for 1.5 h (oxygen glucose deprivation) before a normal medium was used for 24 h (reperfusion).Then,the cells were divided into the cell control group,OGD/R group,OGD/R+Sal A 1,5 and 10 μmol·L-1 group,OGD/R+ML385 group,OGD/R+ML385+Sal A 1,5 and 10μmol·L-1 group and OGD/R+edaravone (Eda,50μmol·L-1) group.After twenty-four hours of culture,the cell survival rate was measured by CCK8 kit.The contents of Interleukin-1β (IL-1β),IL-6,tumor necrosis factor-α(TNF-α),IL-10,IL-4 and transforming growth factor-β(TGF-β) in the cell supernatant were detected by ELISA.Reactive oxygen species (ROS) in cells was detected using the chemical fluo-rescence method.The contents of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD),glutathione peroxidase (GSH-PX) and chloramphenicol acetyltransferase (CAT) in cells were determined with the colorimetric method.Protein expressions of Kelch like ECH-associated protein 1 (Keap1),nuclear factor erythroid-2 related factor 2 (Nrf2),Heme oxygenase-1 (HO-1),NADPH:quinone oxidoreductase-1 (NQO1) and p-nuclear factor kappa-B p65 (p-NF-κB p65) were detected by Western blotting.RESULTS ①Compared with the cell control group,the cell survival rate of the OGD/R group was significantly decreased (P＜0.01).Compared with the OGD/R group,the survival rates of OGD/R+Sal A 1,5 and 10μmol·L-1 groups were significantly increased (P＜0.05,P＜0.01).②Compared with the cell control group,the contents of IL-1β,IL-6 and TNF-α were significantly increased,the contents of IL-10,IL-4 and TGF-β were significantly decreased,the contents of ROS and MDA were significantly increased,and the activities of SOD,CAT and GSH-Px were significantly decreased in the OGD/R group (P＜0.01).Compared with the OGD/R group,the content of IL-6 was significantly decreased,the contents of IL-10,IL-4 and TGF-β were significantly increased,the contents of ROS and MDA were significantly decreased,and the activities of SOD,CAT and GSH-Px were significantly increased in OGD/R+Sal A 1,5 and 10μmol·L-1 and OGD/R+Eda groups (P＜0.05,P＜0.01).③Compared with the cell control group,the protein expression of p-NF-κB P65 in the OGD/R group was significantly increased (P＜0.01).Compared with the OGD/R group,the protein expressions of Keap1 and cytoplasmic Nrf2 were significantly decreased,the expressions of nuclear Nrf2,HO-1 and NQO1 proteins were significantly increased,and the expression of p-NF-κB p65 protein was significantly decreased in OGD/R+Sal A 5 and 10 μmol·L-1 and OGD/R+Eda groups (P＜0.05,P＜0.01).In OGD/R+ML385,OGD/R+ML385+Sal A 1,5 and 10μmol·L-1 groups,the protein expression of Keap1 was significantly increased,the protein expressions of cytoplasmic Nrf2,nuclear Nrf2,HO-1 and NQO1 protein were significantly decreased,and the protein expression of p-NF-κB P65 was significantly increased (P＜0.01).CONCLU-SION Sal A reduces the inflammatory response and oxidative stress of OGD/R injured BV2 cells possi-bly by activating the Keap1/Nrf2 pathway and inhibiting the NF-κB pathway.

Eukaryotic translation initiation factor 4B (eIF4B) plays an important role in mRNA translation initiation, cell survival and proliferation in vitro, but the in vivo function is poorly understood. In this study, via various experimental techniques such as hematoxylin-eosin (HE) staining, flow cytometry, Western blotting, and immunohistochemistry, we investigated the role of eIF4B in mouse embryo development using an eIF4B knockout (KO) mouse model and explored the mechanism. We found that the livers, but not lungs, brain, stomach, or pancreas, derived from eIF4B KO mouse embryos displayed severe pathological changes characterized by enhanced apoptosis and necrosis. Accordingly, high expression of cleaved-caspase 3, and excessive activation of mTOR signaling as evidenced by increased expression and phosphorylation of p70S6K and enhanced phosphorylation of 4EBP1, were observed in mouse embryonic fibroblasts and fetal livers from eIF4B KO mice. These results uncover a critical role of eIF4B in mouse embryo development and provide important insights into the biological functions of eIF4B in vivo.
Animals ; Apoptosis/genetics* ; Caspase 3 ; Eosine Yellowish-(YS) ; Eukaryotic Initiation Factors/metabolism* ; Fibroblasts ; Hematoxylin ; Liver/metabolism* ; Mice ; Ribosomal Protein S6 Kinases, 70-kDa/genetics* ; TOR Serine-Threonine Kinases

Objective To identify the potential prognostic biomarkers of the immune-related genes signature for patients with hepatocellular carcinoma (HCC). Methods Original HCC data were downloaded from TCGA, and the immune activity of each sample was calculated by ssGSEA. HCC samples were divided into high and low immune cell infiltration groups by "GSVA" package and "hclust" package. The ESTIMATE algorithm scored the tumor microenvironment in each HCC sample. The "limma" package and Venn diagram identified effective immune-related genes. Univariate Cox, Lasso regression and multivariate Cox regression analyses were used to explore key genes. The "rms" package was used to create nomograms and draw calibration curves. Results Compared with the high immune cell infiltration group, the tumor purity of the samples in the low immune cell infiltration group was higher, the immune score, ESTIMATE score and stromal score were lower. In the high immune cell infiltration group, the immune components were more abundant, and the expression levels of TIGIT, PD-L1, PD-1, LAG3, TIM-3, CTLA4 and HLA family were higher. Multivariate Cox regression analysis showed that four immune-related genes (S100A9, HMOX1, IL18RAP and FCER1G) were used to construct the prognosis model. Compared with other clinical features, the risk score of this prognostic model was recognized as an independent prognostic factor. Conclusion This study identified the immune-related core genes which may be used in targeted therapy and immunotherapy of HCC.

0bjective To study the angular dependence of optically stimulated luminescent dosimeter (OSLD) with solid phantoms under SSDL radiation level ⁶⁰Co radiation field, and to discuss the possibility of OSLD in volumetric modulated arc therapy (VMAT) and other rotating irradiation dose audit. Methods OSLDs were embedded in the two phantoms with the same size and material, respectively. The phantom 1 was set to make the first OSLD perpendicular to the beam, and the phantom 2 was set to make the second OSLD parallel to the beam. The OSLDs were irradiated at 8 angles: 0°, 45°, 90°, 135°, 180°, −45°, −90, −135°. The counts of the OSLSs were read and the response of each angle which normalized to 0° were calculated. Results When the OSLDs are perpendicular to the beam, the angular response is between −6.76% ～ +1.5%, with the maximum angular dependence at 90° and −90°. When the OSLDs are parallel to the beam, the angular response is between −1.74%～+1.67%, below 2%. Conclusion It is better to correct the sensitivity of dosimeters by Element Correction Factors (ECF) for dose audit. Under the condition of rotating irradiation, OSLD shoud be set parallel to the beam, which can better reduce the influence of angular dependence and facilitate further application research of VMAT dose audit.

Objective:To investigate the risk factors and treatments of recurrent cesarean scar pregnancy (RCSP).Methods:A retrospective case-controlled study was performed in 48 cesarean scar pregnancy (CSP) patients in Shenzhen Maternity & Child Healthcare Hospital from January 2014 to December 2018. Recurrent group consisted of 12 patients with RCSP, while control group consisted of 36 patients experienced single CSP. The information of pregnancy, surgery and other factors of the two groups was compared.Results:Thickness of lower uterine segment in recurrent group was thinner than that in control group [(1.81±1.12) mm vs. (2.80±1.56) mm, P=0.031]. Dilation and curettage (D&C) was the main treatment method, and scar repair was the secondary treatment in these two groups. Patients may receive prophylactic uterine artery embolization (UAE) to prevent catastrophic bleeding during surgery when necessary. Totally 11 of 12 patients were cured in recurrent group, while all of 36 patients were cured in control group. Univariate analysis showed that artificial abortions ≥3 times ( P=0.007), type of CSP and thickness of lower uterine segment <3 mm ( P=0.013) had a significant correlation with the RCSP. Conclusion:Artificial abortions ≥3 times and thickness of lower uterine segment <3 mm were the independent risk factors of RCSP. D&C can be used as the first line treatment of RCSP, and prophylactic UAE should be performed when necessary.

Objective:To investigate the risk factors and treatments of recurrent cesarean scar pregnancy (RCSP).Methods:A retrospective case-controlled study was performed in 48 cesarean scar pregnancy (CSP) patients in Shenzhen Maternity & Child Healthcare Hospital from January 2014 to December 2018. Recurrent group consisted of 12 patients with RCSP, while control group consisted of 36 patients experienced single CSP. The information of pregnancy, surgery and other factors of the two groups was compared.Results:Thickness of lower uterine segment in recurrent group was thinner than that in control group [(1.81±1.12) mm vs. (2.80±1.56) mm, P=0.031]. Dilation and curettage (D&C) was the main treatment method, and scar repair was the secondary treatment in these two groups. Patients may receive prophylactic uterine artery embolization (UAE) to prevent catastrophic bleeding during surgery when necessary. Totally 11 of 12 patients were cured in recurrent group, while all of 36 patients were cured in control group. Univariate analysis showed that artificial abortions ≥3 times ( P=0.007), type of CSP and thickness of lower uterine segment <3 mm ( P=0.013) had a significant correlation with the RCSP. Conclusion:Artificial abortions ≥3 times and thickness of lower uterine segment <3 mm were the independent risk factors of RCSP. D&C can be used as the first line treatment of RCSP, and prophylactic UAE should be performed when necessary.

Objective:To investigate the application value of three-dimensional visualization technology in management of middle hepatic vein (MHV) processing in associating liver partition and portal vein ligation for staged hepatectomy(ALPPS).Methods:The retrospective and descriptive study was conducted. The clinical data of 40 patients with right massive liver cancer or multiple right liver lesions who underwent ALPPS in the First Affiliated Hospital of Guangxi Medical University from November 2017 to August 2019 were collected. There were 34 males and 6 females, aged (44±9)years, with a range from 26 to 64 years. All patients underwent multi-slice computed tomography (CT) plain and enhanced scan of superior abdominal region before operation, and the data were transmitted to the liver visualization analysis software IQQA system with 1.5 mm thin-layer images to complete the three-dimensional reconstruction of the liver and its blood vessels. Patients were performed ALPPS based on results of three-dimensional reconstruction and intraoperative findings. Observation indicators: (1) results of preoperative three-dimensional reconstruction; (2) surgical situations; (3) follow-up. Follow-up was conducted using outpatient examinations and telephone interview to detect postopeartive survival of patients up to March 2020. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were described as M (range). Count data were represented as absolute numbers. Results:(1) Results of preoperative three-dimensional reconstruction: 40 patients underwent three-dimensional reconstruction successfully, of which 37 clearly showed MHV, tumor location and relationship between them, 3 patients showed unclearly MHV and were classified based on two-dimensional images. Of the 40 patients, 12 had MHV classified as type A, 13 as type B, 9 as type C, and 6 as type D. Three-dimensional reconstruction of vessels showed 22 with umbilical veins and 9 with anterior veins. Of the 40 patients, 35 were predicted to preserve MHV, and 5 were predicted to resect MHV. Total estimated liver volume, tumor volume, and reserved liver volume were (1 012±119)cm 3, 600 cm 3(8-2 055 cm 3), (346±80)cm 3. The ratio of future liver remnant to standard liver volume was 34%±8%. (2) Surgical situations : 40 patients underwent the first-stage ALPPS, including 35 with preservation of MHV and 5 with resection of MHV, which was accorded with preoperative prediction. Thirty-four patients underwent the second-stage ALPPS, and 6 patients had failure to receive the second-stage ALPPS due to undificiency future liver remnant. The operation time and volume of intraoperative blood loss for 40 patients undergoing first-stage ALPPS were (350±79)minutes and 300 mL(range, 100-2 600 mL). Three patients received blood transfusion and no perioperative death occurred. There were 24 patients with grade A heptic insufficiency according to criteria of International StudyGroup of Liver Surgery (ISGLS) and 16 patients with grade B heptic insufficiency after the first-stage ALPPS. Twenty-eight patients had grade Ⅰ complications of Clavien-Dindo classification, including 17 with a small pleural effusion, 10 with a small pleural and abdominal effusion, 1 with hypoproteinemia; 8 patients had grade Ⅱ complications of Clavien-Dindo classification, including 5 with pneumonia, 1 with pneumonia combined with pleural and abdominal effusion, 1 with coagulation disorders, 1 with biliary fistula; 3 patients had grade Ⅲ complications of Clavien-Dindo classification, including 2 with pneumothorax and pneumonia, 1 with pneumothorax, pneumonia and coagulation disorders; 1 patient had grade Ⅳ complications of Clavien-Dindo classification as systemic inflammatory response syndrome. All patients with complications were improved after symptomatic treatment, anti infection, transfusion of fresh frozen plasma or drainage. For the 34 patients undergoing the second-stage ALPPS, the operation time and volume of intraoperative blood loss were (320±83)minutes and 500 mL(range, 200-6 000 mL). Twelve patients received blood transfusion. There were 12 patients with grade A heptic insufficiency according to criteria of ISGLS and 22 with grade B heptic insufficiency after the second-stage ALPPS. Eighteen patients had grade Ⅰ complications of Clavien-Dindo classification, including 11 with a small pleural effusion, 7 with a small pleural and abdominal effusion; 12 patients had grade Ⅱ complications of Clavien-Dindo classification, including 4 with pneumonia, 4 with coagulation disorders, 3 with massive abdominal effusion, 1 with biliary fistula; 3 patients had grade Ⅲ complications of Clavien-Dindo classification, including 1 with pneumothorax and pneumonia, 1 with massive pleural effusion, 1 with obstructive jaundice; 1 patient had grade Ⅳ complications of Clavien-Dindo classification as pneumonia and anemia. All patients with complications were improved after symptomatic treatment, anti infection, transfusion of fresh frozen plasma or drainage. (3) Follow-up: 40 patients were followed up for 2-35 months, with a median follow-up time of 17 months. The 6-month, 1-, and 2-year survival cases were 35, 26, 21 cases. Conclusion:Three-dimensional visualization technology can clearly show the MHV classification and its relationship with tumor location, which has an important guiding significance in the decision-making of MHV management in ALPPS.