OBJECTIVE To study the effects of the curcumin derivative bisdemethoxycurcumin （BC） promoting neuronal differentiation of neuroblastoma cells Neuro-2a （N2a） in mice and its mechanism. METHODS The effects of BC （1， 2， 4， 6， 8， 10 μmol/L） on the viability of N2a cells were detected by MTT assay to determine the concentration range of drug treatment. The control group， retinoic acid （RA） group （10 μmol/L） and BC groups （1， 2 and 4 μmol/L） were set up， and the length of neural protrusions of the differentiated cells was measured and the cell differentiation rate was calculated after 48 h and 72 h of culture. Compared with 0 min group， Western blot was used to detect the phosphorylation levels of protein kinase B （Akt）， extracellular- signal regulated kinase 1/2 （ERK1/2）， and p38 mitogen-activated protein kinase （p38） proteins in cells treated by 4 μmol/L BC for 5， 15， 30， 60， 120 min. After intervention with inhibitors LY294002 （LY） and PD98059 （PD）， the effects of BC on Akt and ERK1/2 protein phosphorylation levels and promoting neural differentiation were further validated. RESULTS According to the MTT experiment， the BC concentrations for subsequent induction of cell differentiation were determined to be 1， 2， and 4 μmol/L. After 48 hours of differentiation， compared with the control group， the cell differentiation rate in RA group and BC 1， 2 and 4 μmol/L groups， the length of cellular neural processes wjxhhxx413@163.com in the BC 4 μmol/L group significantly increased （P＜0.05 or P＜0.01）；after inducing differentiation of BC for 72 hours，compared with the control group， the cell differentiation rate and the length of cellular neural processes in the RA group， the cell differentiation rate in the BC 4 μmol/L group， and the length of cellular neural processes in the BC 2 μmol/L group all significantly increased （P＜0.05 or P＜0.01）.Compared with the 0 min group， the phosphorylation levels of Akt， ERK1/2， and p38 proteins in cells of the 5， 15， 30， 60 and 120 min groups increased to varying degrees after treated by 4 μmol/L BC， and some differences were statistically significant （P＜0.05 or P＜0.01）. After adding the inhibitor LY/PD， compared with the BC group， the phosphorylation level of ERK1/2 protein in the PD+BC group cells were significantly reduced （P＜0.01）， and the cell differentiation rates in the LY group， LY+BC group， PD group， and PD+BC group was significantly reduced （P＜0.01）. CONCLUSIONS BC promotes N2a cell differentiation mainly by increasing cell differentiation rate and neural protrusion length. The mechanism may be related to the activation of mitogen-activated protein kinase/ ERK and PI3K/Akt signaling pathways.

Objective:To evaluate the significance of copy number variation (CNV) and metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid (CSF) in the diagnosis of meningeal carcinomatosis (MC).Methods:Ten patients with MC diagnosed in the Department of Neurology of Peking Union Medical College Hospital from March 2022 to June 2022 were consecutively enrolled in this study. The patients were diagnosed according to the criteria of the Chinese expert consensus on the diagnosis of MC by the Chinese Society of Infectious Diseases and Cerebrospinal Fluid Cytology, and the diagnosis of MC was confirmed by CSF cytology. The control group included 10 patients who were diagnosed as autoimmune encephalitis or viral encephalitis. CSF mNGS and CNV analysis were performed simultaneously in all the patients.Results:Of the 10 patients with MC, 6 had lung adenocarcinoma, 4 had breast cancer. CSF mNGS and CNV analysis detected large CNV in 8 of 10 patients with MC, including 4 patients with breast cancer and 4 patients with lung cancer. The results of pathogenic microorganism analysis of CSF mNGS in all the patients were negative. Meanwhile, large CNV was not detected in the control group.Conclusions:CSF CNV can serve as a diagnostic marker for MC. The combination of mNGS and CNV analysis has demonstrated a high positive rate in the diagnosis of MC. The dual-omics analysis of pathogenic microorganisms and CNV has been proposed as a potential strategy to further expand the clinical utility of CSF mNGS in the realm of auxiliary diagnosis.

ObjectiveTo investigate the application of Mengchao Liver Disease-Brain System version 2.0 in clinical diagnosis and treatment. MethodsThis study was conducted among 160 patients who were admitted to the internal medicine and surgical departments from June 9 to 21， 2021， and their data were automatically captured by the intelligent information system of Southeast Big Data Institute of Hepatobiliary Health， Mengchao Hepatobiliary Hospital of Fujian Medical University. The completeness and accuracy of Mengchao Liver Disease-Brain System version 2.0 were evaluated based on the intelligent diagnostic tools such as auxiliary diagnosis of chronic hepatitis B， interpretation of liver fibrosis， staging model of chronic hepatitis B， auxiliary diagnosis of liver cirrhosis， auxiliary staining of liver cirrhosis， auxiliary diagnosis of primary liver cancer， BCLC stage of primary liver cancer， Chinese staging of primary liver cancer， Child-Pugh score， and APRI score. ResultsAll auxiliary diagnostic tools had a complete rate of 94.17% in terms of the extraction of correct key dimensions within the test period. The artificial intelligence report had a structured accuracy of 97.55% in capturing data and an accuracy rate of 91.61% in text processing. ConclusionMengchao Liver Disease-Brain System version 2.0 provides an innovative mode for the construction of big data platform in medical specialties and has a high accuracy as an auxiliary diagnostic tool in clinical diagnosis and treatment.

Objective:To summarize the clinical features, treatments and outcomes of patients with SMARCB1(INI-1)-deficient sinonasal carcinoma (SDSC).Methods:Fifteen patients who were diagnosed as SDSC in Beijing Tongren Hospital from October 2016 to June 2021 were retrieved, including nine males and six females, ranged from 25 to 78 years old. For TNM stage, one case was in stage T2, one case was in stage T3, 13 cases were in stage T4; 13 cases were in stage N0, two cases were in stage N2; 14 cases were in stage M0, one case was in stage M1. The most common paranasal sinus affected by tumor was the ethmoid sinus. Five patients were treated by radical surgical resection combined with postoperative adjuvant therapy, four patients treated by neoadjuvant therapy with surgical resection, three patients treated by surgical resection only, one patient treated by neoadjuvant therapy with concurrent chemoradiotherapy, one patient treated by preoperative radiotherapy with surgery, and one patient received palliative chemotherapy. Immunohistochemical analysis was performed in all cases. The Kaplan-Meier method was used to draw the survival curve, and the Log-rank test was used to compare the difference to 20 undifferentiated carcinoma patients with positive INI-1 expression in the same period.Results:Immunohistochemical analysis showed the complete absence of INI-1 expression in the tumor nuclei in all 15 cases. The follow-up information was available with a median follow-up time of 21 months (3-56 months). The 3-year overall survival rate, disease specific survival rate, disease-free survival rate and metastasis-free survival rate were 58.9%, 58.9%, 36.4% and 31.2%, respectively. Disease-free survival in SDSC patients was significantly lower compared with undifferentiated carcinoma patients with positive INI-1 expression ( HR=2.87,95% CI:0.92~8.91, P=0.043). Cox regression analysis showed that patients with comprehensive treatment based on surgery had a better prognosis than others ( HR=8.61,95% CI:1.38~53.73, P=0.021). Conclusion:SDSC is a highly aggressive malignant tumor with the characteristics of easy recurrence, early metastasis and poor prognosis. INI-1 immunohistochemical analysis is recommended in the pathologically poorly differentiated sinonasal carcinoma. Comprehensive treatment based on radical resection may be the first choice for SDSC patients.

Objective:To investigate the clinical features and prognosis of pediatric autoimmune encephalitis associated with anti-glutamic acid decarboxylase 65 (GAD65) antibody.Methods:Clinical data of 2 patients diagnosed as autoimmune encephalitis associated with anti-GAD65 antibody at Department of Neurology, Beijing Children′s Hospital in 2019 were analyzed retrospectively. A literature search with “anti-GAD65 antibody”“encephalitis”“epilepsy” or “cerebellar ataxia” as key words was conducted at China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform and PubMed (up to January 2020). The clinical features and prognosis of pediatric cases with complete clinical data were retrieved and summarized.Results:Two patients with positive anti-GAD65 antibody of serum and cerebrospinal fluid were both females. The onset age of case 1 was 57 months and her main clinical manifestations were fever and unconsciousness. The cranial magnetic resonance imaging (MRI) showed diffuse T2 weighted imaging (T2WI) abnormal signals, and the electroencephalogram (EEG) showed slow waves. The onset age of case 2 was 80 months and her main clinical manifestations of were recurrent focal seizures, memory loss, and headache. The MRI showed high T2WI signal in bilateral hippocampus, and the EEG showed abnormal discharge involving the temporal area. Both cases were treated with methylprednisolone and intravenous immunoglobulin, the short-term symptoms of them were both improved. They were followed up for 6 months and 1 year respectively, the case 1 recovered completely, and the case 2 still had focal seizures. Six English reports which included 6 cases were retrieved. Together with these 2 cases, a total of 8 cases were analyzed. The clinical symptoms included seizures (6 cases), memory loss (4 cases), loss of consciousness (3 cases), behavioral abnormalities (3 cases), cognitive impairment (2 cases), headache (2 cases), autonomic symptoms (1 case), ataxia (1 case), dysphagia (1 case), and aphasia (1 case). There were 5 cases with cranial MRI abnormalities in the acute phase or sub-acute phase, of whom 3 cases had the limbic system involvement, and 2 cases were mainly had extra limbic area involvement. Three cases had hippocampal atrophy or sclerosis during follow-up. All 8 patients were treated with immunotherapy. After immunotherapy, all patients had short-term improvement. Follow-up for 6 months to 6 years showed that 3 cases with extra limbic encephalitis improved to baseline levels, and 5 limbic encephalitis cases had poor outcomes, including 1 death and 4 cases still had focal epilepsy.Conclusions:Pediatric anti-GAD65 antibody associated autoimmune encephalitis is a rare but treatable disease, including limbic encephalitis and extra limbic encephalitis. The most common clinical manifestations are seizures and memory impairment. Early diagnosis and immunotherapy can improve the symptoms in a short time. But patients with limbic encephalitis often had refractory epilepsy in the chronic phase, and have a poor long-term outcome.

Objective: To analyze the differentially expressed genes related to the chemosensitivity with the TPF regimen for hypopharyngeal squamous cell carcinoma and to measure potential functional targeting genes expressions. Methods: Twenty-nine patients with primary hypopharyngeal cancer who underwent induction chemotherapy with TPF from January 2013 to December 2017 in Beijing Tongren Hospital were enrolled for microarray analysis, including 28 males and 1 female, aged from 43 to 73 years old. Among them, 16 patients were sensitive to chemotherapy while 13 patients were non-sensitive. Illumina Human HT-12 Bead Chip was applied to analyze the gene expressions and online bioinformatics analysis was used to analyze the differentially expressed genes. Reverse transcription and quantitative real-time PCR (RT-qPCR) was used to measure the mRNA expression of potential functional genes of TPF induction chemotherapy in 43 samples, 29 from original patients and 14 from additional patients. Graphpad prism 7.0 software was used for statistical analysis. Results: A total of 1 381 significantly differentially expressed genes were screened out. By GO analysis, up-regulated genes included sequestering in extracellular matrix, chemokine receptor binding and potassium channel regulator activity; down-regulated genes included regulation of angiogenesis, calcium ion binding and natural killer cell activation involved in immune response. With KEGG database analysis, down-regulated pathways included ECM-receptor interaction and peroxisome and up-regulated pathways included Glutathione metabolism and PPAR signaling pathway. The expressions of CD44 and IL-6R were significantly different and appeared biologically significant. CD44 was significantly upregulated in insensitive tissues (0.54±0.06) compared with sensitive tissues (0.33±0.04)(P<0.01). IL-6R was significantly downregulated in insensitive tissues (0.44±0.03) compared with sensitive tissues. (0.68±0.03) (P<0.01). Conclusion CD44 and IL-6R may be potentially functional genes of TPF induction chemotherapy in hypopharyngeal squamous cell carcinoma.

Objective:To evaluate the application of computer-assisted design and three-dimensional printing technique in reconstruction of maxillary defects with individual free fibular flap.Methods:A total of 13 patients, 7 males and 6 females with age from 12 to 55 years old, underwent the reconstruction of maxillary defects after subtotal or total maxillectomy for benign or malignant tumors between January 2016 and December 2018 were reviewed. Ther were 4 cases of subtotal maxillectomy and 9 cases of total maxillectomy. Before operation, osteotomy line was planned on three-dimensional images. A three-dimensional individual resin fibula model based on mirror images of the healthy side maxilla was obtained to fabricate an anatomically adapted osteomyocutaneous fibula free flap using computer-assisted design and forming. Oral, nasal, and eye functions and facial appearance were evaluated.Results:The 13 cases were followed up for 5-40 months, all flaps were alive except one due to flap failure and all cases were tumor free confirmed by CT or MRI. Ten patients could eat normal food without nasal food return, and 3 cases had palatal fistula. Eleven cases showed good speaking functional results. All cases had normal eye positions, no cases with diplopia and loss of vision. With evaluation by VAS, an average score of 8 was obtained, and most of patients were satisfied with their postoperative facial appearances.Conclusion:Reconstruction of maxillary defects by free fibular flap designed by three-dimensional printing and computer-assisted technique not only restored oral, nasal and eye functions, but also showed satisfactory facial appearance.

Objective:To explore the method of functional protection in transoral endoscopic thyroidectomy by vestibular approach.Methods:Retrospective analysis was performed on the case data of 66 patients who underwent transoral endoscopic thyroidectomy by vestibular approach in the Department of Otorhinolaryngology Head and Neck Surgery of Beijing Tongren Hospital from February 2018 to February 2020. There were 11 males and 55 females aged 22-64 years, including 19 patients with benign diseases and 47 patients with malignant diseases. Important neurological functions were protected by a combination of anatomical exposure and intraoperative nerve monitoring. The clinical data and the effect of neurofunctional protection were summarized. SPSS 25.0 software was used for statistical analysis.Results:All operations were successfully completed with none transferred to open surgery. Three weeks after surgery, the numbness and tingling symptoms in the lower jaw and lower lip were basically relieved. There was no permanent mental nerve palsy, no permanent recurrent laryngeal nerve palsy but 2 cases with temporary recurrent laryngeal nerve palsy, no external branch injury of superior laryngeal nerve. There were 3 cases of temporary parathyroid gland dysfunction, 1 case of neck infection, 2 cases of subcutaneous effusion, 1 case of neck skin injury, and 2 cases of postoperative eye conjunctivitis.Conclusion:In transoral endoscopic thyroidectomy by vestibular approach, anatomical exposure can decrease mental nerve injury, and anatomical exposure combined with intraoperative nerve monitoring can protect the functions of the external branches of the superior laryngeal nerve and the recurrent laryngeal nerve.

Objective:To evaluate clinical applications and efficacy of submental artery perforator flap in reconstruction surgery after removal of pharyngeal carcinoma.Methods:A total of 27 patients in the Department of Otorhinolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University were included, 23 males and 4 females with age from 40 to 70 years old, and 17 patients were hypopharyngeal carcinoma (HPC) and 10 patients were oropharyngeal carcinoma (OPC). All patients underwent tumor resection followed by simultaneously reconstruction surgery using submental artery perforator flap between January 2015 and December 2019. Of 5 patients with palatine tonsil cancer, 4 underwent the combined approach of neck and oral resection and 1 with madibulotomy. All 5 patients with tongue base cancer received transhyoid partial glossotomy with or without partial laryngectomy. Sixteen patients with pyriform sinus carcinoma received partial laryngo-pharyngectomy with preservation of laryngeal functions. One patient with posterior hypopharyngeal wall carcinoma had partial pharyngectomy. Prognosis and laryngeal functions were analyzed after reconstruction surgery with submental artery perforator flap in patients with pharyngeal carcinoma.Results:The 27 patients were followed up for 6-66 months, with a median of 13 months, of them 24 patients were alive without recurrence or metastasis, 1 patient died of recurrence, 1 patient died of esophageal carcinoma and 1 patient was alive with the recurrence of tongue base carcinoma. Postoperative complications included flap failure for 1 case, pharyngeal fistula for 1 case, subcutaneous hydrops for 2 cases and lymphatic fistula for 1 case. Total 2 and 3 year survival rates were 92.9% and 88.9%, respectively. Total decanulation rate was 92.6%; decanulation rate and intubation time were 16/17 and 3.5 months in HPC patients; and decanulation rate and intubation time were 9/10 and 2 months in OPC patients. Total oral feeding rate was 92.6% and nasogastric feeding time was 3.5 weeks in HPC patients and 3 weeks in OPC patients.Conclusion:The submental artery perforator flap is an excellent choice for reconstruction surgery after removal of oropharyngeal and hypopharyngeal carcinoma, with good outcomes of laryngeal functions.

Objective:To study the significance of induction chemotherapy and subsequent comprehensive therapy for overall survival rate (OS) and larynx dysfunction-free survival rate (LDFS) in patients with advanced hypopharyngeal carcinoma.Methods:Patients who met the inclusion criteria with the diagnoses of advanced hypopharyngeal carcinoma between 2011 and 2017 received 2 or 3 cycles of TPF regimen induction chemotherapy. Patients who attained complete response (CR) received radical chemotherapy. Patients who attained partial response (PR) and the reduction of tumor volume was more than 70% were defined as large PR and received concurrent chemoradiotherapy. When the tumor volume reduction of PR patients was less than 70%, they were defined as small PR. (CR+large PR) group was defined as effective group. Patients who did not reach CR and large PR were defined as uneffective group and underwent radical surgery and received adjuvant radiotherapy as appropriate after the surgery. The end points of the study were OS, progression-free survival (PFS) and LDFS. Chi-square (χ 2) test was used for correlation analysis. Survival analysis was performed by the Kaplan-Meier method with a Log-rank test. Cox proportional hazards model was used for univariate and multivariate survival analysis. Results:A total of 260 patients were enrolled in the study. The follow-up period ranged from 5 to 83 months, with an average of 24.7 months. The 3-year and 5-year OS rate was 46.0% and 32.6%, respectively. The 3-year and 5-year PFS rate was 41.0% and 26.6%, respectively. The 3-year and 5-year LDFS rate was 37.9% and 24.8%, respectively. Poor outcome of induction chemotherapy, advanced N stage, strong positive Ki-67 immunohistochemistry (all P<0.001) were negative prognostic factors. The advanced clinical stage was positively related to the poor outcome of induction chemotherapy ( P=0.015). There was no significant difference in OS and PFS between the large PR group and the small PR group (all P>0.005). Conclusion:TPF regimen induction chemotherapy and subsequent comprehensive therapy for patients with advanced hypopharyngeal carcinoma may improve the quality of life of patients, with high OS rate and LDFS rate.