Glioblastoma(GBM)is a lethal cancer with limited therapeutic options.Dendritic cell(DC)-based cancer vaccines provide a promising approach for GBM treatment.Clinical studies suggest that other immu-notherapeutic agents may be combined with DC vaccines to further enhance antitumor activity.Here,we report a GBM case with combination immunotherapy consisting of DC vaccines,anti-programmed death-1(anti-PD-1)and poly I:C as well as the chemotherapeutic agent cyclophosphamide that was integrated with standard chemoradiation therapy,and the patient remained disease-free for 69 months.The patient received DC vaccines loaded with multiple forms of tumor antigens,including mRNA-tumor associated antigens(TAA),mRNA-neoantigens,and hypochlorous acid(HOCl)-oxidized tumor lysates.Furthermore,mRNA-TAAAs were modified with a novel TriVac technology that fuses TAAs with a destabilization domain and inserts TAAs into full-length lysosomal associated membrane protein-1 to enhance major histo-compatibility complex(MHC)class Ⅰ and Ⅱ antigen presentation.The treatment consisted of 42 DC cancer vaccine infusions,26 anti-PD-1 antibody nivolumab administrations and 126 poly I:C injections for DC infusions.The patient also received 28 doses of cyclophosphamide for depletion of regulatory T cells.No immunotherapy-related adverse events were observed during the treatment.Robust antitumor CD4+and CD8+T-cell responses were detected.The patient remains free of disease progression.This is the first case report on the combination of the above three agents to treat glioblastoma patients.Our results suggest that integrated combination immunotherapy is safe and feasible for long-term treatment in this patient.A large-scale trial to validate these findings is warranted.

Objective: To explore the therapeutic effects of transoral robotic surgery (TORS) and traditional surgical modes in oropharyngeal squamous cell carcinoma (OPSCC). Methods: The clinicopathological data of patients with oropharyngeal squamous cell carcinoma treated at Sun Yat-sen University Cancer Center from 2010 to 2018 were retrospectively analyzed. 135 cases were treated with traditional surgery (non-TORS group), while 52 cases were treated with TORS (TORS group). The prognosis of the two groups of patients were analyzed by Kaplan-Meier method and Log rank test, the influencing factors were analyzed by Cox regression model. Results: The 2-year overall survival (OS, 94.2%) and 2-year progression-free survival (PFS, 93.8%) of patients in the TORS group were better than those in the non-TORS group (71.4% and 71.4%, respectively, P<0.05). The 2-year OS (93.3%) and 2-year PFS (92.8%) of TORS group patients in T1-2 stage were better than those of non-TORS group (73.1% and 72.8%, respectively, P<0.05). The 2-year OS (95.8%) and 2-year PFS (95.2%) of patients with stage Ⅰ to Ⅱ in the TORS group were not significantly different from those in the non-TORS group (84.1% and 83.9%, respectively, P>0.05). The 2-year OS (92.9%) and 2-year PFS rate (92.7%) of patients with stage Ⅲ to Ⅳ in the TORS group were better than those in the non-TORS group (64.7% and 63.9%, respectively, P<0.05). The 2-year OS (94.4%) of HPV-positive patients in the TORS group was not significantly different from that in the non-TORS group (83.3%, P=0.222). The 2-year OS of HPV-negative patients in the TORS group (94.1%) was significantly different from that in the non-TORS group (43.7%, P<0.001). HPV status was an independent prognostic factor (P=0.008). Conclusions: TORS has a better prognosis in the treatment of oropharyngeal squamous cell carcinoma compared with the traditional treatment methods. The patients with T1-T2 can achieve better survival benefits after TORS treatment. The HPV-positive OPSCC patients has a better prognosis than that of HPV-negative OPSCC patients, and regardless of HPV status, OPSCC patients in the TORS group could obtain a better survival prognosis.
Head and Neck Neoplasms ; Humans ; Oropharyngeal Neoplasms/surgery* ; Papillomavirus Infections/complications* ; Retrospective Studies ; Robotic Surgical Procedures/methods* ; Squamous Cell Carcinoma of Head and Neck/surgery*

Objective: To investigate the functional changes of key gut microbiota (GM) that produce lipopolysaccharide (LPS) in atrial fibrillation (AF) patients and to explore their potential role in the pathogenesis of AF. Methods: This was a prospective cross-sectional study. Patients with AF admitted to Beijing Chaoyang Hospital of Capital Medical University were enrolled from March 2016 to December 2018. Subjects with matched genetic backgrounds undergoing physical examination during the same period were selected as controls. Clinical baseline data and fecal samples were collected. Bacterial DNA was extracted and metagenomic sequencing was performed by using Illumina Novaseq. Based on metagenomic data, the relative abundances of KEGG Orthology (KO), enzymatic genes and species that harbored enzymatic genes were acquired. The key features were selected via the least absolute shrinkage and selection operator (LASSO) analysis. The role of GM-derived LPS biosynthetic feature in the development of AF was assessed by receiver operating characteristic (ROC) curve, partial least squares structural equation modeling (PLS-SEM) and logistic regression analysis. Results: Fifty nonvalvular AF patients (mean age: 66.0 (57.0, 71.3), 32 males(64%)) were enrolled as AF group. Fifty individuals (mean age 55.0 (50.5, 57.5), 41 males(82%)) were recruited as controls. Compared with the controls, AF patients showed a marked difference in the GM genes underlying LPS-biosynthesis, including 20 potential LPS-synthesis KO, 7 LPS-biosynthesis enzymatic genes and 89 species that were assigned as taxa harbored nine LPS-enzymatic genes. LASSO regression analysis showed that 5 KO, 3 enzymatic genes and 9 species could be selected to construct the KO, enzyme and species scoring system. Genes enriched in AF group included 2 KO (K02851 and K00972), 3 enzymatic genes (LpxH, LpxC and LpxK) and 7 species (Intestinibacter bartlettii、Ruminococcus sp. JC304、Coprococcus catus、uncultured Eubacterium sp.、Eubacterium sp. CAG:251、Anaerostipes hadrus、Dorea longicatena). ROC curve analysis revealed the predictive capacity of differential GM-derived LPS signatures to distinguish AF patients in terms of above KO, enzymatic and species scores: area under curve (AUC)=0.957, 95%CI: 0.918-0.995, AUC=0.940, 95%CI 0.889-0.991, AUC=0.972, 95%CI 0.948-0.997. PLS-SEM showed that changes in lipopolysaccharide-producing bacteria could be involved in the pathogenesis of AF. The key KO mediated 35.17% of the total effect of key bacteria on AF. After incorporating the clinical factors of AF, the KO score was positively associated with the significantly increased risk of AF (OR<0.001, 95%CI:<0.001-0.021, P<0.001). Conclusion: Microbes involved in LPS synthesis are enriched in the gut of AF patients, accompanied with up-regulated LPS synthesis function by encoding the LPS-enzymatic biosynthesis gene.
Aged ; Atrial Fibrillation/complications* ; Cross-Sectional Studies ; Gastrointestinal Microbiome ; Humans ; Lipopolysaccharides ; Male ; Middle Aged ; Prospective Studies

Objective: To evaluate the indications, safety, feasibility, and surgical technique for patients with head and neck cancers undergoing transoral robotic retropharyngeal lymph node (RPLN) dissection. Methods: The current study enrolled 12 consecutive head and neck cancer patients (seven males and four females) who underwent transoral robotic RPLN dissection with the da Vinci surgical robotic system at the Sun Yat-sen University Cancer Center from May 2019 to July 2020. Seven patients were diagnosed as nasopharyngeal carcinoma with RPLN metastasis after initial treatments, 4 patients were diagnosed as thyroid carcinoma with RPLN metastasis after initial treatments, and one patient was diagnosed as oropharyngeal carcinoma with RPLN metastasis before initial treatments. The operation procedure and duration time, intraoperative blood loss volume and complications, nasogastric feeding tube dependence, tracheostomy dependence, postoperative complications, and hospitalization time were recorded and analyzed. Results: All patients were successfully treated by transoral robotic dissection of the metastatic RPLNs, none of which was converted to open surgery. RPLNs were completely resected in 10 patients, and partly resected in 2 patients (both were nasopharyngeal carcinoma patients). The mean number of RPLN dissected was 1.7. The operation duration time and intraoperative blood loss volume were (191.3±101.1) min and (150.0±86.6) ml, respectively. There was no severe intraoperative complication such as massive haemorrhage or adjacent organ injury during surgery. Nasogastric tube use was required in all patients with (17.1±10.6) days of dependence, while tracheotomy was performed in 8 patients with (11.6±10.7) days of dependence. The postoperative hospitalization stay was (8.5±5.7) days. Postoperative complications occurred in 4 patients, including 2 of retropharyngeal incision and 2 of dysphagia. During a follow-up of (6.5±5.1) months, disease-free progression was observed in all patients, 10 patients were disease-free survival and other 2 patients were survival with tumor burden. Conclusions: The transoral robotic RPLN dissection is safety and feasible. Compared with the traditional open surgical approach, it is less traumatic and safer, has fewer complications and good clinical application potentiality. The indications for transoral robotic RPLN dissection include thyroid carcinoma, oropharyngeal carcinoma, and some selected nasopharyngeal carcinoma and other head and neck cancers. Metastatic RPLNs from some nasopharyngeal carcinoma with incomplete capsule, unclear border and adhesion to the surrounding vessels are not suitable for transoral robotic RPLN dissection.
Blood Loss, Surgical ; Female ; Head and Neck Neoplasms/pathology* ; Humans ; Lymph Node Excision/methods* ; Lymph Nodes/pathology* ; Male ; Nasopharyngeal Carcinoma/pathology* ; Nasopharyngeal Neoplasms/surgery* ; Neck Dissection/methods* ; Postoperative Complications/surgery* ; Robotic Surgical Procedures/methods* ; Thyroid Neoplasms/pathology*

Pancreatic ductal adenocarcinoma (PDAC) is characterized by the highest mortality among carcinomas. The pathogenesis of PDAC requires elevated autophagy, inhibition of which using hydroxychloroquine has shown promise. However, current realization is impeded by its suboptimal use and unpredictable toxicity. Attempts to identify novel autophagy-modulating agents from already approved drugs offer a rapid and accessible approach. Here, using a patient-derived organoid model, we performed a comparative analysis of therapeutic responses among various antimalarial/fungal/parasitic/viral agents, through which econazole (ECON), an antifungal compound, emerged as the top candidate. Further testing in cell-line and xenograft models of PDAC validated this activity, which occurred as a direct consequence of dysfunctional autophagy. More specifically, ECON boosted autophagy initiation but blocked lysosome biogenesis. RNA sequencing analysis revealed that this autophagic induction was largely attributed to the altered expression of activation transcription factor 3 (ATF3). Increased nuclear import of ATF3 and its transcriptional repression of inhibitor of differentiation-1 (ID-1) led to inactivation of the AKT/mammalian target of rapamycin (mTOR) pathway, thus giving rise to autophagosome accumulation in PDAC cells. The magnitude of the increase in autophagosomes was sufficient to elicit ER stress-mediated apoptosis. Furthermore, ECON, as an autophagy inhibitor, exhibited synergistic effects with trametinib on PDAC. This study provides direct preclinical and experimental evidence for the therapeutic efficacy of ECON in PDAC treatment and reveals a mechanism whereby ECON inhibits PDAC growth.

OBJECTIVES: The metabolomics technique of LC-MS/MS combined with data analysis was used to detect changes and differences in metabolic profiles in the vitreous humor of early rat carcasses found in water, and to explore the feasibility of its use for early postmortem submersion interval (PMSI) estimation and the cause of death determination. METHODS: The experimental model was established in natural lake water with 100 SD rats were randomly divided into a drowning group (n=50) and a postmortem (CO₂ suffocation) immediately submersion group (n=50). Vitreous humor was extracted from 10 rats in each group at 0, 6, 12, 18 and 24 h postmortem for metabolomics analyses, of which 8 were used as the training set to build the model, and 2 were used as test set. PCA and PLS multivariate statistical analysis were performed to explore the differences in metabolic profiles among PMSI and causes of death in the training set samples. Then random forest (RF) algorithm was used to screen several biomarkers to establish a model. RESULTS: PCA and PLS analysis showed that the metabolic profiles had time regularity, but no differences were found among different causes of death. Thirteen small molecule biomarkers with good temporal correlation were selected by RF algorithm. A simple PMSI estimation model was constructed based on this indicator set, and the data of the test samples showed the mean absolute error (MAE) of the model was 0.847 h. CONCLUSIONS The 13 metabolic markers screened in the vitreous humor of rat corpses in water had good correlations with the early PMSI. The simplified PMSI estimation model constructed by RF can be used to estimate the PMSI. Additionally, the metabolic profiles of vitreous humor cannot be used for early identification of cause of death in water carcasses.
Animals ; Biomarkers/metabolism* ; Cadaver ; Chromatography, Liquid ; Immersion ; Postmortem Changes ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Vitreous Body/metabolism* ; Water/metabolism*

OBJECTIVE: To clarify the mechanism by which LINC01285 regulates proliferation and migration of colorectal cancer (CRC) cells and the clinical implications. METHODS: We analyzed the expression of LINC01285 in CRC tissues and normal tissues using data from Starbase public database. We also examined the expression levels of LINC01285 in 70 pairs of CRC and adjacent tissue samples collected from our center and in different CRC cell lines using RT-qPCR, and analyzed the correlation of LINC01285 expression with the clinicopathological parameters and tumor-free survival time of the patients. In CRC cell lines (SW620 and HT-29), the changes in cell proliferation, apoptosis, metastasis and epithelial-mesenchymal transition (EMT) phenotype following LINC01285 knockdown were analyzed using CCK-8 assay, flow cytometry, Transwell assay and Western blotting. RESULTS: The TCGA-COAD transcriptome sequencing data obtained from the Starbasev3.0 public database revealed a significantly higher expression level of LINC01285 in CRC tissues than in adjacent tissues (P=0.00016), which was verified by RT-qPCR results of the clinical samples (P=0.0002). In CRC patients, the expression level of LINC01285 was closely correlated with histological differentiation of the tumor (P=0.036), T classification (P=0.000), lymph node metastasis (P=0.001), TNM stage (P=0.000), Duke stage (P=0.009) and relapse-free survival (P=0.0102). In SW620 and HT-29 cells, which expressed significantly higher levels of LINC01285 than normal colorectal mucosal cells (P < 0.001), LINC01285 knockdown significantly inhibited cell proliferation (P < 0.001), increased early apoptosis, late apoptosis and total apoptosis rates (P < 0.05), suppressed cell migration and invasion (P < 0.001), upregulated the expression of E-cadherin (P < 0.001), and downregulated the expression of N-cadherin (P < 0.001). CONCLUSION The expression level of LINC01285, which modulates the EMT pathway to regulate the proliferation, apoptosis and metastasis of CRC cells, is closely correlated with the prognosis of CRC patients.
Humans ; Epithelial-Mesenchymal Transition ; Neoplasm Recurrence, Local ; HT29 Cells ; Cell Proliferation ; Colorectal Neoplasms

Objective: To evaluate the safety, efficacy and feasibility of transoral robotic surgery (TORS) for parapharyngeal space (PPS) neoplasms. Methods: We collected data from 7 patients with PPS neoplasm who received TORS in Sun Yat-sen University Cancer Center between May 2017 and November 2020, and patients' clinical and pathological characteristics were analysed. There were 2 men and 5 women with age ranged from 35 to 76 years. Among them, 2 patients underwent secondary surgery, 2 patients required combined transcervical approach to complete surgery, and 1 patient was suspected of ipsilateral cervical lymph node metastasis and scheduled for diagnostic TORS. The preoperative tumor size, operation time, intraoperative blood loss, postoperative bleeding, dyspnea, neurological impairment, feeding time and postoperative hospital stay were analyzed. SPSS 24.0 was used to analyze the data. Results: TORS was performed successfully with complete removal of tumors in all 7 cases. Among 6 patients with curative TORS, 5 patients received TORS with postoperative diagnoses of neurogenic tumors and 1 patient underwent TORS combined transcervical approach with postoperative disgnosis of recurrent pleomorphic adenoma; no intraoperative tumor rupture occurred; the intraoperative blood loss was 20-200 ml with a median of 40 ml; the operation time was 65.0-238.0 min with a median of 77.5 min; the oral feeding time was 3-6 days with a median of 3 days; and the postoperative hospital stay was 4.2±1.6 days. One patient presented with neck swelling 3 days after surgery, but this symptom relieved 3 days later after treatments with antibiotic, hemostasis and detumescence. One patient received diagnostic TORS, as intraoperative pathology indicating a recurrent pleomorphic adenoma, then the neoplasm got completely resected through transcervical-transparotid approach. None of 7 patients manifested with airway obstruction, bleeding or nerve injury symptoms after operation. All patients were followed for 2 to 44 months, no local recurrence or distant metastasis was found. Conclusions: TORS is a safe, effective and feasible treatment for selected PPS neoplasms, with less cosmetic impact, less trauma and blood loss, few postoperative complications, enhanced postoperative recovery and short hospital stay.
Adult ; Aged ; Female ; Head and Neck Neoplasms ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Parapharyngeal Space ; Pharyngeal Neoplasms ; Retrospective Studies ; Robotic Surgical Procedures

ObjectiveTo investigate the effect of imperatorin on liver damage， peroxidative stress and immune response in young rats with type I diabetes induced by streptozotocin （STZ）. MethodsTotally 50 BALB/c juvenile mice were equally divided into healthy control group， model group， model plus+3 mg/kg Imperatorin group， model+6 mg/kg Imperatorin group， and model+12 mg/kg Imperatorin group. Glucose tolerance test was used to detect fasting blood glucose， serum insulin and obesity index of mice in each group. Automatic biochemical analyzer was used to determine serum HDL， LDL， TG， TC content. HE staining was used to observe the degree of liver damage. TUNEL staining was used to observe cell apoptosis. SOD and MDA levels were detected by Kit， Box/Bcl-2， cleaved cas3/cas3， c-Myc expression levels in mitochondria by Western blot， and serum IL-6， TNF-α， IL-1β levels by ELISA. ResultsCompared with the model group， 12 mg/kg Imperatorin significantly reduced fasting blood glucose ［mean difference 5.3 mmol/L， 95% confidence interval （4.5， 6.1） mmol/L］， serum insulin ［mean difference 13.2 μU/L， 95% confidence interval （10.6， 15.8） μU/L］ and obesity index ［average difference 2.3 kg/m²， 95% confidence interval （2.1， 2.5） kg/m²］ all decreased. The content of SOD increased （P<0.05）， while the content of MDA was reduced （P<0.05）. The expression level of Bax/Bcl-2， and cleaved cas3/cas3 was down-regulated （P<0.05）， while the level of c-Myc was up-regulated （P<0.05）， and the content of IL-6， TNF-α and IL-1β was reduced （P<0.05）. ConclusionImperatorin can alleviate the liver damage， peroxidation and immune response in young rats with STZ-induced type I diabetes.

Objective:To observe the effect of Dredging Correcting Manipulation on cblC methylmalonic aciduria (MMA). Methods:From October, 2017 to October, 2018, 72 children with cblC MMA combined with growth retardation were divided into control group (n = 36) and experimental group (n = 36) according to the consent of their parents. The control group accepted routine medicine, and the experimental group received Dredging Correcting Manipulation in addition. The Griffiths Development Scale-Chinese version (GDS-C) was used to evaluate the two groups before and after treatment. At the same time, body length, body mass and head circumference were measured. Results:Six cases in the control group and five cases in the experimental group were dropped out. There was no significant difference in the development quotients of GDS-C in gross movement, personal and social, hearing and speech, hand-eye coordination, operation and total quotient between two groups before treatment (P > 0.05). After treatment, all the development quotients increased in both groups (t > 6.110, P < 0.001), and the development quotients of GDS-C in gross movement, personal and social, hand-eye coordination and total quotient were higher in the experimental group than in the control group (t > 2.154, P < 0.05), as well as the body length (t = 2.027, P < 0.05). Conclusion:Dredging Correcting Manipulation can promote the neuropsychological and physical development of children with cblC MMA combined with retardation.