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.

Soft tissue sarcoma (STS) is a group of rare malignant tumors originating from mesenchymal tissue, with a high degree of malignancy and a wide range of pathological subtypes. The prognosis varies among different subtypes, and treatment increasingly relies on selecting appropriate treatment methods for different subtypes. Surgical treatment is still the main treatment method at present, and the development of immune and targeted therapy also brings new hope for the treatment of soft tissue sarcoma. Immune checkpoint inhibitors, oncolytic viruses and T cell therapy have shown well safety and efficacy in clinical trials. Targeted drugs such as trabectedin and lenvatinib have changed the treatment pattern of soft tissue sarcoma. Currently, chemotherapy based on doxorubicin and ifosfamide is still the first line treatment for patients with advanced soft tissue sarcoma who have distant metastasis. However, the adverse reactions of doxorubicin limit its application in elderly patients, and trofosfamide has shown good efficacy and safety as an alternative in clinical trials. The efficacy of postoperative radiotherapy has been confirmed, which can reduce the local recurrence rate after surgical resection of soft tissue sarcoma. In summary, multimodal comprehensive treatment has become the main strategy for the treatment of soft tissue sarcoma. The combination of different treatment methods can generate synergistic effects and help patients obtain more clinical benefits, such as the combination of doxorubicin and immune checkpoint inhibitors, and the combination of antiangiogenic drugs and chemotherapy drugs. At the 2023 annual meeting of the American Society of Clinical Oncology (ASCO), oncologists from all over the world reported many researches related to the treatment of soft tissue sarcoma. This article aims to review the new progress in the treatment of soft tissue sarcoma in the 2023 annual meeting of ASCO.

This study aims to construct C57/B6-L and A549 cell lines that stably overexpress circu-lar RNA LAMP3(circLAMP3),laying the foundation for further research on the biological func-tions of circLAMP3.Total RNA was extracted and reverse transcripted into cDNA from C57/B6-L and A549 cells to amplify the full-length sequence of circLAMP3.Then,the fragments of cir-cLAMP3 were ligated into pLC5-ciR vector to obtain pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 recombinant plasmids.The lentiviruses expressing circLAMP3 were packaged on tran-sient transfected HEK293T cells.C57/B6-L and A549 cells were infected with lentiviruses to gen-erate cell lines overexpressing circLAMP3 through puromycin screening.To verify the overexpres-sion efficiency of circLAMP3 of cell lines,we performed the fluorescence microscopy,PCR amplifi-cation,quantitative PCR(qPCR),and Sanger sequencing experiments.The results indicated that the overexpression plasmids of pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 were suc-cessfully constructed.Strong green fluorescence was observed under a fluorescence microscopy.C57/B6-L and A549 cell lines showed a significant increase in the expression of circLAMP3 by PCR and qPCR methods.Sanger sequencing results showed that the junction site of circLAMP3 was correct.This study successfully constructed C57/B6-L and A549 cell lines overexpressing circLAMP3,providing biomaterials for further exploration of the biological function of circLAMP3 in influenza virus replication.

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 explore the effect of 1-methyltryptophan (1-MT) on lipopolysaccharide (LPS)-induced permeability and apoptosis of human umbilical vein endothelial cells (HUVECs).Methods:HUVECs were treated with phosphate buffer saline (PBS, control group), 1 μg/mL LPS (LPS group), and LPS combined with 1 mmol/L 1-MT (1-MT group). The expression levels of the p120 concatemer (p120ctn), vascular endothelial (VE) cadherin, caspase-3, and DNA repair enzyme polyadenylate ribose polymerase-1 (PARP) after incubation at 8 h were detected using Western blot. The concentrations of kynurenine (Kyn) after incubation at 2, 4, 6, and 8 h were measured by high-performance liquid chromatography, and indoleamine2, 3-dioxygenase (IDO) activity was calculated. Comparisons among groups were performed using the LSD- t test. Results:Compared with the control group, the expression of caspase-3 [(74.01±7.91)% vs (157.14±7.63)%, P<0.01] and the concentration of Kyn were significantly up-regulated, while the expression of p120ctn [(49.12±2.15)% vs (37.61±1.80)%, P<0.01], VE-cadherin [(107.70±7.01)% vs (90.66±2.58)%, P=0.027], and PARP-1 [(67.95± 3.08)% vs (57.93±5.26)%, P=0.038] were significantly down-regulated, and IDO activity was significantly increased in the LPS group ( P<0.05). Compared with the LPS group, the expression of caspase-3 [(157.14±7.63)% vs (110.74±7.89)%, P<0.01] was significantly down-regulated, while the expression of p120ctn [(37.61±1.80)% vs (47.19±0.82)%, P<0.01], VE-cadherin [(90.66±2.58)% vs (107.27±9.89)%, P=0.029], and PARP-1 [(57.93±5.26)% vs (74.12±4.90)%, P=0.005] were significantly up-regulated, and the activity of IDO was significantly decreased over time in the 1-MT group ( P<0.05). No significant differences were observed between the PBS and 1-MT groups in the protein levels of p120ctn, VE-cadherin, and PARP-1 protein as well as Kyn concentration and IDO activity ( P>0.05), while the expression of caspase-3 was increased in 1-MT group ( P=0.001). Conclusions:LPS aggravates the permeability of HUVECs, which can be reversed by 1-MT via inhibiting IDO activity and reducing Kyn concentrations. Moreover, 1-MT can also reduce apoptosis, which may be via increasing the expression of PARP-1 and reducing the expression of caspase-3, thus protecting endothelial cells.

Objective To explore the application value of laparoscopic pedicled omentum acquisition technique in laparoscopic breast-conserving surgery of breast carcinoma.Methods From November 2016 to November 2017,there were 11 patients with breast carcinoma underwent laparoscopic breast-conserving surgery and breast plastic surgery with laparoscopic pedicled omentum acquisition technique in Beijing Friendship Hospital,Capital Medical University,and retrospectivly analyzed the clinical data of these patients.The operation time,intraoperative blood loss,drainage tube time,length of incision,surgical complications,postoperative cosmetic effect and tumor recurrence were recorded and analyzed.Results In 11 patients,the average of the total operation time was 186 min,the average intraoperative blood loss was 22 ml,the average drainage tube time was 3.1 d,and the average total length of the incision was 5.2 cm.There was no severe complication after surgery but only one patient had an increase of belching.The postoperative cosmetic results were excellent with the mean score was 14.7 (the total score was 15) along with getting full score in "shape of breast" and "scar of the incision".There was no locoregional recurrence or distant metastases.Conclusion The combination of laparoscopic breast-conserving surgery of breast carcinoma and the laparoscopic pedicled omentum acquisition technique assure the recovery of the breast shape safely and effectively,also provide better postoperative cosmetic results.

Objective: To analyze the distribution and associated factors of high-risk genotypes of HPV in cervical infection among women in Shenzhen. Methods: The information on sociodemographic characteristics and HPV genotypes of HPV-positive women who participated cervical screening test from January 2014 to December 2016 was downloaded from Shenzhen Maternity and Child Healthcare Management Information System. According to the pathogenicity, the high-risk HPV genotypes were divided into 15 types including HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66 and 68; and there were 6 low-risk genotypes including HPV 6, 11, 42, 43, 44, and 81. Chi-square tests were applied to compare the proportions of high-risk HPV infection among women who had different sociodemographic characteristics. A non-conditional logistic regression model was used to analyze the associated factors for high-risk HPV infection. Results: In total, all HIV positives received HPV genotyping, with an average age of (38.08±9.38) years old. There were 9 979 (93.9%) high-risk and 645 (6.1%) low-risk HPV infections. The proportions of HPV infections for high-risk type in each year were 91.5%, 93.8%, and 95.6%, increasing with the screening years (χ²=54.79, P<0.001). Multivariate logistic regression analysis showed that compared with women younger than 25 years old, women in other age groups (at age 26 to 30 years, 31 to 35 years, 36 to 40 years, 41 to 45 years, and 50 years or older) had increased risks of high-risk HPV infection, with OR (95%CI) of 1.67 (1.20-2.31), 1.49 (1.09-2.03), 1.71 (1.23-2.37), 1.65 (1.19-2.31), and 1.84 (1.26-2.67), respectively; compared with the married, single women had a decreased risk of high-risk HPV infection (OR (95%CI): 0.71 (0.50-1.00)); women received HPV testing in 2015 and 2016 showed higher risk of high-risk HPV infection than those in 2014 (OR (95%CI): 1.43 (1.17-1.74) and 2.03 (1.68-2.46)). The 5 most common HPV genotypes were HPV52 (25.1%, 2 670 cases), followed by HPV16 (19.2%, 2 041 cases), HPV58 (13.3%, 1 413 cases), HPV18 (9.9%, 1 048 cases), and HPV51 (9.3%, 993 cases). Conclusion Age, marital status, and screening year were associated with high-risk HPV infections. Besides HPV16 and HPV18, the prevention and control on HPV infections for HPV52, HPV58, and HPV51 should be prioritized in Shenzhen area.

Objective: To analyze the epidemiological genotype features of human papillomavirus (HPV) in cervical infection and their risks for cervical precancers among women in Shenzhen area. Methods: A total of 2 717 individuals ranging in age from 30~59 years were recruited in 18 community health centers of Shenzhen city from March 1 to June 15, 2015 by a cluster sampling method. The results of genotype of HPV, liquid-based cytology (LBC), colposcopy and pathology were analyzed. The clinical sensitivity and specificity as well as positive (PPV) and negative (NPV) predictive values of the combination of different HPV genotype in screening the cervical intraepithelial neoplasia (CIN) 2 and above were estimated. Results: The HPV infection rate in Shenzhen area was 15.9% (432/2 717). The most common HPV genotype was HPV52 (22.9%), followed by HPV16 (12.7%), HPV53 (10.0%), HPV51 (8.6%) and HPV58 (8.1%). Compared with HPV16/18 genotyping, HPV33/16 genotyping had a higher sensitivity (57.1% vs. 42.9%, P<0.05) and an analogous specificity (87.3% vs. 86.9%, P>0.05) in predicting CIN2+ . The sensitivity of combination of HPV33/16 genotyping and low grade squamous intraepithelial lesion (LSIL) positive tested by LBC in predicting CIN2+ was 75.0%, significantly higher than 64.3% of atypical squamous cells of undetermined significance (ASC-US) positive tested by LBC alone (P<0.05). The specificities of these two methods mentioned above in predicting CIN2+ were 83.5% and 89.2%, respectively, without statistical difference (P>0.05). Conclusions Women infected by HPV have distinct risks for CIN2+ according to different high-risk HPV genotypes. The top five risks were HPV 33, 16, 58, 56, and 68. HPV-positive women triaged by LBC LSIL+ combined with HPV33/16 genotyping may be a potential strategy for cervical cancer screening in developed urban area.