Objective:To explore the short-term outcomes of 3D-printing stand-alone artificial vertebral body(AVB)in the surgical procedure of anterior cervical corpectomy and fusion(ACCF).Methods:Following the proposal of IDEAL(idea,development,exploration,assessment,and long-term follow-up)framework,we designed and conducted this single-armed,retrospective cohort study.The patients with cervical spondylotic myelopathy were recruited,and these patients exclusively received the surgical procedure of single-level ACCF in our single center.After the process of corpectomy,the size was tailored using different trials and the most suitable stand-alone AVB was then implanted.This AVB was manufactured by the fashion of 3D-printing.Two pairs of screws were inserted in an inclined way into the adjacent vertebral bodies,to stabilize the AVB.The participants were regularly followed-up after the operation.Their clinical data were thoroughly reviewed.We assessed the neurological status according to Japanese Orthopedic Association(JOA)scale.We determined the fusion based on imaging examination six months after the operation.The recorded clinical data were analyzed using specific software and they presented in suitable styles.Paired t test was employed in comparison analysis.Results:In total,there were eleven patients being recruited eventually.The patients were all followed up over six months after the operation.The mean age of the cohort was(57.2±10.2)years.The mean operation time was(76.1±23.1)min and the median bleeding volume was 150(100,200)mL.The postoperative course was uneventful for all the cases.Dysphagia,emergent hematoma,and deterioration of neurological func-tion did not occur.Mean JOA scores were 13.2±2.2 before the operation and 16.3±0.8 at the final follow-up,which were significantly different(P＜0.001).The mean recovery rate of neurological func-tion was 85.9％.By comparing the imaging examinations postoperatively and six months after the opera-tion,we found that the average subsidence length was(1.2±1.1)mm,and that there was only one ca-ses(9.1％)of the severe subsidence(＞3 mm).We observed significant improvement of cervical lor-dosis after the operation(P=0.013).All the cases obtained solid fusion.Conclusion:3D-printing stand-alone AVB presented favorable short-term outcome in one-level ACCF in this study.The fusion rate of this zero-profile prosthesis was satisfactory and the complication rate was relatively low.

[Objective] To analyze factors influencing the postoperative progression-free survival (PFS) in patients with renal cell carcinoma (RCC), construct a nomogram model for predicting PFS, and compare it with other predictive models. [Methods] A retrospective analysis was conducted on the general and clinical data of 263 RCC patients who underwent surgery at the Department of Urology, the Second Affiliated Hospital of Xi'an Jiaotong University, during Apr.2014 and Nov.2021.Patients were divided into the progression group (n=34) and non-progression group (n=229). The data of the two groups were analyzed to identify prognostic variables associated with PFS, and a nomogram model was constructed.The performance of this model was compared with that of the University of California, Los Angeles Integrated Staging System (UISS) score, tumor staging, tumor size, tumor pathological grade, and tumor necrosis scoring system (SSIGN score), and Leibovich score by plotting receiver operating characteristic (ROC) curve and calculating the area under the curve (AUC). Calibration curve of the nomogram was used to validate the model's performance, and K-fold cross-validation was employed to assess its external validity. [Results] Multivariate Cox regression analysis revealed that age (HR=2.255, 95%CI: 1.032-4.926), T stage (HR=5.766, 95%CI: 2.351-14.142), pathological grade (HR=3.100, 95%CI: 1.445-6.651), and pathological necrosis (HR=2.656, 95%CI: 1.253-5.629) were independent risk factors of PFS (P<0.05). The nomogram model based on these four independent variables had AUCs (95%CI) of 0.750 (0.630-0.870), 0.803 (0.705-0.902), and 0.847 (0.757-0.937) for 1, 3, and 5 years, respectively, which were higher than those of UISS score, SSIGN score, and Leibovich score.The calibration curve of the nomogram showed good consistency between predicted and actual probabilities.In K-fold cross-validation, the average AUCs of the nomogram at 1, 3, and 5 years were 0.761, 0.808, and 0.842, indicating good external validity of the nomogram. [Conclusion] The nomogram based on age, T stage, pathological grade and pathological necrosis can accurately predict the risk of postoperative PFS in RCC patients at 1, 3, and 5 years, which can aid clinicians in the early identification of high-risk progression.

Heme/metabolism* ; Cryoelectron Microscopy ; Membrane Transport Proteins ; Escherichia coli Proteins/metabolism*

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins

【Objective】 To investigate the effects of preoperative lipid metabolism level on the postoperative prognosis of non-muscular invasive bladder cancer (NMIBC). 【Methods】 Clinical data of NMIBC patients who underwent surgical treatment in our hospital during Mar.2014 and May 2021 were retrospectively analyzed. Based on receiver operating characteristic (ROC) curve, the optimal cutoff values of all lipid metabolism indicators were determined and patients were classified accordingly. The independent risk factors for postoperative recurrence were identified with Cox regression model. The survival was analyzed with Kaplan-Meier, and recurrence-free survival (RFS) was compared using log-rank tests. A recurrence risk prediction model was established based on the high-density lipoprotein (HDL) and other clinic pathological factors and the accuracy of prediction was evaluated with the area under the ROC curve (AUC). 【Results】 Cox multivariate analysis showed HDL, tumor number, tumor size and histological grade were independent risk factors for recurrence (P<0.05). Kaplan-Meier analysis showed that RFS was significantly longer in the high-HDL group than in the low-HDL group (P<0.001). Incorporating HDL, tumor number, tumor size, histological grade, and tumor stage into the recurrence risk model, the AUC was 0.706, and internal cross validation showed the AUC was 0.711. 【Conclusion】 Preoperative HDL is an independent risk factor affecting the RFS of patients with NMIBC, and combining it with clinic pathological factors will improve the prediction of tumor recurrence.

【Objective】 To construct an easy-to-use individual survival prognostic tool based on competing risk analyses to predict the risk of 1-, 2- and 3- year recurrence for patients with non-muscle invasive bladder cancer (NMIBC). 【Methods】 The follow-up data of 419 NMIBC patients were obtained. The patients were randomly divided into training cohort (n=293) and validation cohort (n=126). The variables included age at diagnosis, sex, history of smoking, tumor number, tumor size, histolo-gic grade, pathological stage, and bladder perfusion drug. The cumulative incidence function (CIF) of recurrence was estimated using all variables in the training cohort and potential prognostic variables were determined with Gray’s test. The Fine-Gray subdistribution proportional hazard approach was used as a multivariate competitive risk analysis to identify independent pro-gnostic variables. A competing risk nomogram was developed to predict the recurrence. The performance of the competing risk model was evaluated with the area under the receiver operating characteristic curve (AUC), calibration curve, and Brier score. 【Results】 Five independent prognostic factors including age, number of tumors, tumor size, histologic grade and pathological stage were used to construct the competing risk model. In the validation cohort, the AUC of 1-, 2- and 3- year recurrence were 0.895 (95%CI: 0.831-0.959), 0.861(95%CI: 0.774-0.948) and 0.827(95%CI: 0.721-0.934), respectively, indicating that the model had a high predictive performance. 【Conclusion】 We successfully constructed a competing risk model to predict the risk of 1-, 2- and 3-year recurrence for NMIBC patients. It may help clinicians to improve the postoperative management of patients.

Halomonas can grow on diverse carbon sources. As it can be used for unsterile fermentation under high-salt conditions, it has been applied as a chassis for next-generation industrial biotechnology. Short-chain volatile fatty acids, including acetate, propionate, and butyrate, can be prepared from biomass and are expected to be novel carbon sources for microbial fermentation. Halomonas sp. TD01 and TD08 were subjected to shaking culture with 10-50 g/L butyrate, and they were found to effectively synthesize poly-3-hydroxybutyrate with butyrate as the carbon source. The highest yield of poly-3-hydroxybutyrate was achieved at butyrate concentration of 20 g/L (9.12 g/L and 7.37 g/L, respectively). Butyrate at the concentration > 20 g/L inhibited cell growth, and the yield of poly-3-hydroxybutyrate decreased to < 4 g/L when butyrate concentration was 50 g/L. Moreover, Halomonas sp. TD08 can accumulate the copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate by using propionate and butyrate as carbon sources. However, propionate was toxic to cells. To be specific, when 2 g/L propionate and 20 g/L butyrate were simultaneously provided, cell dry weight and polymer titer were 0.83 g/L and 0.15 g/L, respectively. The addition of glycerol significantly improved cell growth and boosted the copolymer titer to 3.95 g/L, with 3-hydroxyvalerate monomer content of 8.76 mol%. Short-chain volatile fatty acids would be promising carbon sources for the production of polyhydroxyalkanoates by Halomonas.
Butyrates ; Carbon ; Fatty Acids, Volatile ; Halomonas ; Polyhydroxyalkanoates ; Propionates

Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors of the head and neck region. NPC has the characteristics of insidious onset, strong invasiveness and early lymph node metastasis. A variety of signaling pathways play a role in the invasion and metastasis of nasopharyngeal carcinoma, but the specific mechanism has not yet been fully elucidated. Recent studies have found that DNA methylation of nasopharyngeal carcinoma-related genes can affect the invasion and metastasis of nasopharyngeal carcinoma through a variety of signaling pathways including Wnt/β-catenin, PI3K/AKT and MAPK signaling pathways. This article reviews the specific mechanism of DNA methylation affecting the invasion and metastasis of nasopharyngeal carcinoma through the above-mentioned signaling pathways.

BACKGROUND: Lung cancer is a malignant with high incidence and mortality and adenocarcinoma is among the most popular subtypes. Epidermal growth factor receptor (EGFR) mutation is one of the most important driver mutations for lung adenocarcinoma and EGFR-tyrosine kinase inhibitor (TKI) will benefit those patients with sensitive EGFR mutations. Recently, immune checkpoint inhibitor (ICI) therapy, provide a new breakthrough treatment for lung cancer patients. Whereas immunotherapy as an emerging treatment does not benefit patients with EGFR mutations, for which mechanistic studies are poorly defined and focused on the link of EGFR mutations and programmed cell death-ligand 1 (PD-L1) expression, we speculate that the different immune microenvironment associated with the two classes of patients. METHODS: Lung adenocarcinoma datasets were collected from the Cancer Genome Atlas (TCGA) database, and clinical information and gene expression profiles were downloaded. The immune related lymphocyte infiltration in TCGA database were generated through timer 2.0 GSEA was used to analyze the difference of pathway expression between EGFR mutant patients and wild type patients. RESULTS: EGFR mutation was more frequently among women and never smokers. Immunoinfiltration analysis showed that patients with EGFR mutation tends to have more tumor associated fibroblasts, common myeloid progenitor cells, hematopoietic stem cells, effector CD4⁺ T cells and natural killer T cells infiltration, and less memory B cells, naïve B cells, plasma B cells, plasmacytoid dendritic cells, memory CD4⁺ T cells, CD4⁺ helper T cells 2, naive CD8⁺ T cells, CD8⁺ T cells and central memory CD8⁺ T cells infiltration. Moreover, patients with more infiltration of CD8⁺ T cells, natural killer T cells, memory B cells and hematopoietic stem cells, tends have better prognosis (Log-rank test, P=0.017, 0.0093, 0.018, 0.016). However, the patients with more CD4⁺ T th2 infiltration in the tumor tends to have worse prognosis (Log-rank test, P=0.016). Furthermore, the results of gene set enrichment analysis showed that compared with the lung adenocarcinoma patients with EGFR wild type, the three pathways positive regulation of natural killer (NK) cell-mediated immune response to tumor cells, NK cell activation involved in immune response, and NK cell-mediated immune response to tumor cells related to natural killer cells in patients with EGFR mutation were down regulated, while the pathway the positive regulation of cytokine secretion involved in immune response was up-regulated in EGFR mutation patients. CONCLUSIONS The tumour microenvironment of patients with EGFR mutations lacks potent tumour killing effector cells and appears dysfunctional with effector cells. This may be a potential reason for the poor efficacy of immunotherapy in patients with EGFR mutations.

OBJECTIVE：To provide reference f or the qualit y sta ndard establishment of Amaranthus retroflexus. METHODS ： Taking 7 batches of A. retroflexus medicinal materials as the research object ，the appearance properties of the medicinal materials were investigated ，and the microscopic characteristics of the medicinal powders were observed. TLC method was adopted to qualitatively identify rutin ，valine and leucine in A. retroflexus medicinal materials. According to the relevant methods of the 2015 edition of Chinese Pharmacopoeia （part Ⅳ），water content ，total ash content ，acid-insoluble ash content and water-soluble extract content were determined. HPLC method was used to determine the content of rutin in the medicinal material of A. retroflexus . The determination was performed on Agilent 5 TC-C18（2）column with mobile phase consisted of methanol- 0.3％ phosphoric acid solution（40∶60，V/V），at the flow rate of 1.0 mL/min. The detection wavelength was set at 358 nm，and the column temperature was 30 ℃. The sample size was 10 μL. RESULTS：The appearance and microstructure characteristics of the medicinal materials were consistent with the existing description. The identification results of TLC meth od showed that 7 batches of medicinal materials and each reference substance （rutin，valine，leucine）showed spots of the same color at the same position. The moisture content of 7 batches of A. retroflexus medicinal materials was 7.43％-8.72％，the total ash content was 11.82％-13.78％，the acid-insoluble ash content was 0.15％-0.55％，and the water-soluble extract content was 17.27％-24.74％. The linear range of rutin was 10-200 μg/mL（R 2＝1.000 0）. RSDs of precision test ，stability test （24 h）and repeatability test were all less than 2.0％ （n＝6）. The average recovery rates of rutin were 99.14％，97.98％ and 98.80％ in low ，medium and high concentration of samples，and RSDs were 0.97％，0.95％，0.96％（n＝3）. The contents of rutin in 7 batches of A. retrophylla were 0.314-1.102 mg/g. CONCLUSIONS：In this study ，character observation ，microscopic identification ，moisture content ，total ash content ，acid- insoluble ash content and water-soluble extract content of A. retroflexus are investigated ；TLC method was established for qualitative identification of leucine ，valine and rutin in A. retroflexus ，and the HPLC method was established for content determination of rutin. It provides reference for the quality standard establishment of A. retroflexus .