Objective To explore the safety of minimally invasive esophagectomy (MIE) with three-field lymphadenectomy (3-FL) for esophageal squamous cell carcinoma (ESCC) by comparing the short-term outcomes between the 3-FL and the two-field lymphadenectomy (2-FL) in MIE. Methods The clinical data of patients with ESCC who underwent minimally invasive McKeown esophagectomy in our hospital from July 2015 to March 2022 were collected retrospectively. Patients were divided into a 3-FL group and a 2-FL group according to lymph node dissection method. And the clinical outcomes and postoperative complications were compared between the two groups. Results A total of 257 patients with ESCC were included in this study. There were 211 males and 46 females with an average age of 62.2±8.1 years. There were 109 patients in the 3-FL group and 148 patients in the 2-FL group. The operation time of the 3-FL group was about 20 minutes longer than that of the 2-FL group (P<0.001). There was no statistical difference between the two groups in the intraoperatve blood loss (P=0.376). More lymph nodes (P<0.001) and also more positive lymph nodes (P=0.003) were obtained in the 3-FL group than in the 2-FL group, and there was a statistical difference in the pathological N stage between the two groups (P<0.001). But there was no statistical difference in the incidence of anastomotic leak (P=0.667), chyle leak (P=0.421), recurrent laryngeal nerve injury (P=0.081), pulmonary complications (P=0.601), pneumonia (P=0.061), cardiac complications (P=0.383), overall complications (P=0.147) or Clavien-Dindo grading (P=0.152) between the two groups. Conclusion MIE 3-FL can improve the efficiency of lymph node dissection and the accuracy of tumor lymph node staging, but it does not increase the postoperative complications, which is worthy of clinical application.

Objective To explore the effects of transurethral en-blocresection of bladder tumor(TeURBT)on urination function,serum chitinase protein 40(YKL-40),bladder specific nuclear matrix protein-1(BLCA-1),and p-oxyfossase-1(PON-1)levels in patients with non-muscle invasive bladder cancer(NMIBC).Methods 74 patients with NMIBC admitted to our Hospital from January 2019 to May 2022 were divided into TeURBT group and transurethral resection of bladder tumor(TURBT)group by random lottery,with 37 cases in each group.The effective tumor clearance rate,surgery and urination function indexes were compared between the two groups.Blood samples were collected before surgery,3 months,6 months and 12 months after surgery to determine the levels of serum YKL-40,BLCA-1 and PON-1,and the recurrence rate was analyzed at 12 months of follow-up.Results The effective resection rate of bladder tumor was 100.00％in both groups.The operative time in TeURBT group was significantly longer than that in TURBT group(P＜0.05),and the intraoperative blood loss,catheter indwelling/bladder irrigation/hospitalization time in TeURBT group were significantly less than those in TURBT group(P＜0.05).Urinary output and maximum urinary flow per second in both groups significantly increased on postoperative day 14(P＜0.05),but there was no statistically significant difference in the above indexes between the two groups on postoperative day 14(P＞0.05).The levels of serum YKL-40 and BLCA-1 in the two groups at 3,6 and 12 months after surgery significantly decreased(P＜0.05),while the level of PON-1 significantly increased(P＜0.05).There were no statistically significant differences in serum YKL-40 and PON-1 levels between the two groups at 3 and 6 months after surgery(P＞0.05),while serum YKL-40 and BLCA-1 levels in the TeURBT group were significantly lower than those in the TURBT group at 12 months after surgery(P＜0.05),and PON-1 levels were significantly higher than those in the TURBT group(P＜0.05).The stages in the TeURBT group could be accurately diagnosed after operation,and the postoperative pathological stages were Ta stage in 24 cases and T1 stage in 13 cases.The total incidence of postoperative complications in TeURBT group and cumulative incidence of recurrence in 1-year follow-up were 5.40％,which was significantly lower than 24.32％and 21.62％in TURBT group(P＜0.05).Conclusion The effective resection rate of TeURBT and TURBT in the treatment of NMIBC tumor is similar,which can effectively improve the urination function of patients,downregulation of serum YKL-40 and BLCA-1 levels,and up-regulation of PON-1 levels.Compared with TURBT,TeURBT has advantages of less intraoperative blood loss,fewer postoperative complications,faster recovery,and lower recurrence rate.

[This corrects the article DOI: 10.1016/j.apsb.2021.08.015.].

Osteosarcoma is a kind of bone tumor with highly proliferative and invasive properties, a high incidence of pulmonary metastasis and a poor prognosis. Chemotherapy is the mainstay of treatment for osteosarcoma. Currently, there are no molecular targeted drugs approved for osteosarcoma treatment, particularly effective drugs for osteosarcoma with pulmonary metastases. It has been reported that fibroblast activation protein alpha (FAPα) is upregulated in osteosarcoma and critically associated with osteosarcoma progression and metastasis, demonstrating that FAPα-targeted agents might be a promising therapeutic strategy for osteosarcoma. In the present study, we reported that the FAPα-activated vinblastine prodrug Z-GP-DAVLBH exhibited potent antitumor activities against FAPα-positive osteosarcoma cells in vitro and in vivo. Z-GP-DAVLBH inhibited the growth and induced the apoptosis of osteosarcoma cells. Importantly, it also decreased the migration and invasion capacities and reversed epithelial-mesenchymal transition (EMT) of osteosarcoma cells in vitro and suppressed pulmonary metastasis of osteosarcoma xenografts in vivo. Mechanistically, Z-GP-DAVLBH suppressed the AXL/AKT/GSK-3β/β-catenin pathway, leading to inhibition of the growth and metastatic spread of osteosarcoma cells. These findings demonstrate that Z-GP-DAVLBH is a promising agent for the treatment of FAPα-positive osteosarcoma, particularly osteosarcoma with pulmonary metastases.

Objective:To analyze the factors influencing radiation-induced liver injury after receiving Cyberknife stereotactic radiotherapy in patients with primary hepatocellular carcinoma.Methods:278 cases with primary hepatocellular carcinoma from July 2016 to April 2019 were prospectively enrolled. Stereotactic radiosurgery with a prescription dose of 48-55gy/5-8 times were given. Liver function, coagulation function, Child-Pugh score, and liver imaging changes were dynamically observed before and after treatment to evaluate the occurrence of radiation-induced liver injury. Logistic regression model was used to analyze the factors influencing radiation-induced liver injury.Results:Among 278 cases, 3 cases of tumor progression were excluded, and a total of 275 cases were included for analysis. The overall survival rate after 8 months of treatment was 100%. Among them, 22 cases were diagnosed as radiation-induced liver injury, with an incidence rate of 8%, and all cases were recovered after symptomatic treatment. Multivariate analysis result suggested that the peripheral white blood cell count was factors influencing the occurrence of radiation-induced liver injury.Conclusion:Cyberknife stereotactic radiotherapy has a low incidence of radiation-induced liver injury in patients with liver cancer, and it is a relatively safe treatment method. Patients with low peripheral white blood cell counts before treatment should be closely monitored for early detection and treatment.

Objective:To analyze the survival benefits and treatment related toxic effects of simultaneous integrated boost intensity-modulated radiotherapy (SIB-RT) for non-operative esophageal squamous cell carcinoma patients.Methods:The data of 2 132 ESCC patients who were not suitable for surgery or rejected operation, and underwent radical radiotherapy from 2002 to 2016 in 10 hospitals of Jing-Jin-Ji Esophageal and Esophagogastric Cancer Radiotherapy Oncology Group (3JECROG) were analyzed. Among them, 518 (24.3%) cases underwent SIB (SIB group) and 1 614 (75.7%) cases did not receive SIB (No-SIB group). The two groups were matched with 1∶2 according to propensity score matching (PSM) method (caliper value=0.02). After PSM, 515 patients in SIB group and 977 patients in No-SIB group were enrolled. Prognosis and treatment related adverse effects of these two groups were compared and the independent prognostic factor were analyzed.Results:The median follow-up time was 61.7 months. Prior to PSM, the 1-, 3-, and 5-years overall survival (OS) rates of SIB group were 72.2%, 42.8%, 35.5%, while of No-SIB group were 74.3%, 41.4%, 31.9%, respectively ( P=0.549). After PSM, the 1-, 3-, and 5-years OS rates of the two groups were 72.5%, 43.4%, 36.4% and 75.3%, 41.7%, 31.6%, respectively ( P=0.690). The univariate survival analysis of samples after PSM showed that the lesion location, length, T stage, N stage, TNM stage, simultaneous chemoradiotherapy, gross tumor volume (GTV) and underwent SIB-RT or not were significantly associated with the prognosis of advanced esophageal carcinoma patients who underwent radical radiotherapy ( P<0.05). Cox model multivariate regression analysis showed lesion location, TNM stage, GTV and simultaneous chemoradiotherapy were independent prognostic factors of advanced esophageal carcinoma patients who underwent radical radiotherapy ( P<0.05). Stratified analysis showed that, in the patients whose GTV volume≤50 cm 3, the median survival time of SIB and No-SIB group was 34.7 and 30.3 months ( P=0.155), respectively. In the patients whose GTV volume>50 cm 3, the median survival time of SIB and No-SIB group was 16.1 and 20.1 months ( P=0.218). The incidence of radiation esophagitis and radiation pneumonitis above Grade 3 in SIB group were 4.3% and 2.5%, significantly lower than 13.1% and 11% of No-SIB group ( P<0.001). Conclusions:The survival benefit of SIB-RT in patients with locally advanced esophageal carcinoma is not inferior to non-SIB-RT, but without more adverse reactions, and shortens the treatment time. SIB-RT can be used as one option of the radical radiotherapy for locally advanced esophageal cancer.

Objective:To analyze the survival benefits and treatment related toxic effects of simultaneous integrated boost intensity-modulated radiotherapy (SIB-RT) for non-operative esophageal squamous cell carcinoma patients.Methods:The data of 2 132 ESCC patients who were not suitable for surgery or rejected operation, and underwent radical radiotherapy from 2002 to 2016 in 10 hospitals of Jing-Jin-Ji Esophageal and Esophagogastric Cancer Radiotherapy Oncology Group (3JECROG) were analyzed. Among them, 518 (24.3%) cases underwent SIB (SIB group) and 1 614 (75.7%) cases did not receive SIB (No-SIB group). The two groups were matched with 1∶2 according to propensity score matching (PSM) method (caliper value=0.02). After PSM, 515 patients in SIB group and 977 patients in No-SIB group were enrolled. Prognosis and treatment related adverse effects of these two groups were compared and the independent prognostic factor were analyzed.Results:The median follow-up time was 61.7 months. Prior to PSM, the 1-, 3-, and 5-years overall survival (OS) rates of SIB group were 72.2%, 42.8%, 35.5%, while of No-SIB group were 74.3%, 41.4%, 31.9%, respectively ( P=0.549). After PSM, the 1-, 3-, and 5-years OS rates of the two groups were 72.5%, 43.4%, 36.4% and 75.3%, 41.7%, 31.6%, respectively ( P=0.690). The univariate survival analysis of samples after PSM showed that the lesion location, length, T stage, N stage, TNM stage, simultaneous chemoradiotherapy, gross tumor volume (GTV) and underwent SIB-RT or not were significantly associated with the prognosis of advanced esophageal carcinoma patients who underwent radical radiotherapy ( P<0.05). Cox model multivariate regression analysis showed lesion location, TNM stage, GTV and simultaneous chemoradiotherapy were independent prognostic factors of advanced esophageal carcinoma patients who underwent radical radiotherapy ( P<0.05). Stratified analysis showed that, in the patients whose GTV volume≤50 cm 3, the median survival time of SIB and No-SIB group was 34.7 and 30.3 months ( P=0.155), respectively. In the patients whose GTV volume>50 cm 3, the median survival time of SIB and No-SIB group was 16.1 and 20.1 months ( P=0.218). The incidence of radiation esophagitis and radiation pneumonitis above Grade 3 in SIB group were 4.3% and 2.5%, significantly lower than 13.1% and 11% of No-SIB group ( P<0.001). Conclusions:The survival benefit of SIB-RT in patients with locally advanced esophageal carcinoma is not inferior to non-SIB-RT, but without more adverse reactions, and shortens the treatment time. SIB-RT can be used as one option of the radical radiotherapy for locally advanced esophageal cancer.

Objective:To investigate whether idebenone can improve behavioral disorders in mice with Parkinson disease (PD) by increasing PHB2 mediated mitophagy.Methods:In the first small experiment, thirty mice were randomly divided into normal control group, model group and treatment group according to the random number table method, with 10 animals in each group.The aim of this study was to observe the effect of idebenone on the behavior of Parkinson disease model mice. In the second experiment, 20 mice were randomly divided into blank control group, MPTP group, shRNA-PHB2 group and shRNA-PHB2+ MPTP group, with 5 mice in each group. The changes of tyrosine dehydrogenase (TH) in brain tissue were detected by immunofluorescence assay. In the third experiment, 30 mice were randomly divided into blank control group, shRNA-PHB2 group, MPTP+ idebenone group, shRNA-PHB2+ MPTP group, shRNA-PHB2+ idebenone group and shRNA-PHB2+ MPTP+ idebenone group, with 5 animals in each group. The aim of this study was to investigate the effect of idebenone on mitochondrial autophagy in mouse brain.C57BL-6 mice were intraperitoneally injected with MPTP to establish the animal model of chronic PD. Then 200 mg / kg idebenone was given by gavage for 21 days. And the expression of PHB2 in brain was inhibited by microinjection of adeno-associated virus 9 (AAV9) shRNA inhibin 2(PHB2) into lateral ventricle. The behavioral changes of the PD mice were detected by Morris water maze, and the changes of tyrosine dehydrogenase (TH) induced by inhibiting PHB2 were detected by immunohistochemistry. The protein expression of LC3 and PHB2 in substantia nigra of midbrain was detected by Western blot.The data were analyzed by GraphPad 7.0 and SPSS 22.0.Results:(1) In the water maze test data of the first small experiment, the repeated measurement ANOVA showed that the group-time interaction effects of latency of mice from 1 to 7 days were significant ( Ftime×group=20.51, P<0.05). Simple effect analysis showed that on the 5th, 6th and 7th day, the incubation period of the treatment group was significantly shortened (all P<0.05). Univariate analysis of variance showed that on the 7th day of the test, the differences between the control group and the model group, the model group and the treatment group, the control group and the treatment group were all statistically significant( t=-49.95, -21.81, 28.14; all P<0.01). In the third small experiment, repeated measurement analysis of variance showed that the interaction between time and group was significant ( Ftime×group=42.11, P<0.01). Simple effect analysis showed that compared with MPTP+ idebenone group, the latency of shRNA-PHB2+ MPTP+ idebenone group was significantly prolonged (all P<0.05). There were no significant difference between shRNA-PHB2+ MPTP+ idebenone group and shRNA-PHB2+ MPTP group except the 4th day ( P<0.05). On the 7th day, compared with MPTP+ idebenone group, the residence time of shRNA-PHB2+ MPTP+ idebenone group was significantly increased ( t=-34.36, P<0.001), but there was no significant difference between shRNA-PHB2+ MPTP group and shRNA-PHB2+ MPTP+ idebenone group ( t=2.94, P>0.05). (2)The results of immunofluorescence experiment showed that the relative expression of TH in the control group, model group, shRNA-PHB2 group and shRNA-PHB2+ MPTP group were (41.03±3.01), (24.20±4.18), (38.39±3.31) and (13.12±2.65), respectively. Compared with the control group, the expression of TH in the midbrain of the MPTP group was significantly down-regulated, the difference was statistically significant( t=7.98, P<0.01). Compared with the MPTP group, the expression of TH in shRNA-PHB2 group was down regulated ( t=-6.73, P<0.05). (3) Western blot results showed that the relative expression of LC3 in midbrain tissue of control group, shRNA-PHB2 group, MPTP+ idebenone group, shRNA-PHB2+ MPTP group, shRNA-PHB2+ idebenone group and shRNA-PHB2+ MPTP+ idebenone group were (0.86±0.07), (0.77±0.08), (0.42±0.05), (0.21±0.05), (0.66±0.09) and (0.27±0.07). The relative expression of PHB2 were (1.13±0.14), (0.56±0.11), (1.08±0.14), (0.27±0.07), (0.68±0.14) and (0.24±0.10). Compared with MPTP+ idebenone group, the relative expression of LC3 and PHB2 in shRNA-PHB2+ MPTP+ idebenone group was significantly decreased ( F=1.96, P<0.01). Conclusion:Idebenone can increase the level of mitophagy in PD mice through PHB2, thus improving the behavioral disorder.

Objective:To investigate the effect and mechanism of exosomes derived from human umbilical cord mesenchymal stem cells (hUC-MSC) in repair of tendon injury in rats.Methods:The hUC-MSC were cultured and the surface markers were identified by flow cytometry. The cells were induced to differentiate into osteoblasts, chondroblasts and adipocytes using a specific media. Meanwhile, the exosomes were isolated from the cell supernatant using exosome separation columns, and were identified by transmission electron microscopy, PKH67 staining and Western blot. A total of 40 Wistar rats were used to establish the Achilles tendon injury model by surgical resection. The rats were divided into hUC-MSC group (Group A) (with 100 μg exosome injected at the injured site) and control group (Group B) (with 250 μl normal saline injected at the injured site) according to the random number table, with 20 rats per group. The expressions of transforming growth factor β (TGF-β), bone morphogenetic protein (BMP-2), vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF-2), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the tendon tissues of both groups were detected using q-PCR, Western blot and immunofluorescence assay at 4 weeks following injection. The expression of collagen III in the injured tissues of both groups was detected by immunohiestochemistry.Results:The isolated and cultured hUC-MSC presented fusiform under an inverted microscope. After osteogenic differentiation, the cells exhibited a cube nodular structure, and the Alizarin red staining was positive. After adipogenic differentiation, the fat was observed inside the cells, which was red by oil red O staining. After chondroblast differentiation, the cells secreted a large amount of glycosaminoglycans, and a strong positive was revealed by Alisin blue staining. The hUC-MSC-derived exosomes showed round disc shape with a depressed internal structure under a transmission electron microscope, which was verified via PKH67 staining. The Western blot analysis showed high expressions of motility-related protein-1 (CD9) and lysosomal associated membrane protein 3 (CD63). The q-PCR test revealed that the mRNA expressions of TGF-β (4.887±0.767), BMP-2 (3.079±0.150), VEGF (3.108±0.508) and FGF-2 (4.211±0.522) in Group A were markedly higher than those in Group B (1.000±0.062, 0.918±0.129, 1.004±0.103, 1.010±0.169, respectively) ( P<0.01), and that the mRNA expression of IL-1β (0.697±0.037) and TNF-α (0.793±0.021) in Group A was markedly lower than those in Group B (1.004±0.089 and 1.006±0.015, respectively) ( P<0.01). The Western blot analysis revealed that the protein expressions of TGF-β (1.434±0.041), BMP-2 (1.798±0.177), VEGF (1.552±0.113) and FGF-2 (1.357±0.039) in Group A were markedly higher than those in Group B (1.002±0.032, 0.992±0.068, 1.007±0.070, 0.994±0.051) ( P<0.01), and that the protein expressions of IL-1β (0.705±0.016) and TNF-α (0.840±0.045) in Group A was markedly lower than those in Group B (1.000±0.016, 1.003±0.040) ( P<0.01). The immunofluorescence revealed that the positive expression rates of TGF-β and VEGF in Group A were not significantly different from those in Group B ( P>0.05). However, the positive expression rates of BMP-2 (2.278±0.208) and FGF-2 (4.656±0.106) in Group A were markedly higher than those in Group B (0.315±0.101, 1.661±0.110) ( P<0.05 or 0.01), and the positive expression rates of IL-1β (1.677±0.947) and TNF-α (1.520±0.088) in Group A were greatly lower than those in Group B (4.296±0.291, 2.373±0.273, respectively) ( P<0.01). In Group A, the tendon collagen fibers were arranged regularly and tightly, with relatively significant expression of collagen III; while the tendon collagen fibers in Group B were distributed loosely, accompanying broken scarlike healing. Conclusion:The hUC-MSC-derived exosomes can prompt the repair of the injured tendon tissues, which may be associated with the function in up-regulating the expressions of growth factors including TGF-β, BMP-2, VEGF and FGF-2, enhancing the expression of collagen III and inhibiting the expression of the inflammatory cytokines including IL-1β and TNF-α.

Objective:To evaluate the effect of definitive radiotherapy with different doses on overall survival (OS) and identify the prognostic factors of patients with non-metastatic esophageal squamous cell carcinoma (ESCC).Methods:Clinical data of 2 344 ESCC patients treated with definitive radiotherapy (RT) alone or chemoradiotherapy from 2002 to 2016 in 10 hospitals were collected and analyzed retrospectively. After the propensity score matching (PSM)(1 to 2 ratio), all patients were divided into the low-dose group (equivalent dose in 2 Gy fractions, EQD 2Gy<60 Gy; n=303) and high-dose group (EQD 2Gy≥60 Gy; n=606) based on the dose of radiation. Survival analysis was conducted by Kaplan- Meier method. Multivariate prognostic analysis was performed by Cox′s regression model. Results:The median follow-up time was 59.6 months. After the PSM, the 1-, 3- and 5-year overall survival (OS) rate was 66.5%, 34.7%, 27.2% in the low-dose group, 72.9%, 41.7% and 34.7% in the high-dose group, respectively ( P=0.018). The 1-, 3-and 5-year progression-free survival rate was 52.2%, 27.2%, 23.1% in the low-dose group, 58.3%, 38.1% and 33.9% in the high-dose group, respectively ( P=0.001). The outcomes of univariate analysis indicated that cervical/upper esophagus location, early (stage Ⅱ) AJCC clinical stage, node negative status, tumor length ≤5 cm, receiving intensity-modulated radiation therapy (IMRT), receiving concurrent chemotherapy and EQD 2Gy≥60 Gy were closely associated with better OS (all P<0.05). Multivariable analysis demonstrated that tumor location, regional lymph node metastasis, concurrent chemotherapy and EQD 2Gy were the independent prognostic factors for OS (all P<0.05). Conclusion:Three-dimensional conformal or IMRT with EQD 2Gy≥60 Gy yields favorable survival outcomes for patients with locally advanced ESCC.