Search

Xinwei Li,1,* Yongfei Fan,2,* Mingyue Tang,1 Huiyuan Li,1 Yue Zhang,1 Jiaqi Mi,1 Yanyan Wang,1 Menglin Zhao,1 Zishu Wang,1 Fang Su1 1Department of Medical Oncology, First Affiliated Hospital of Bengbu Medical College, Bengbu, 233004, People’s Republic of China; 2Department of Thoracic Surgery, The Affiliated Changzhou No. 2 People’s Hospital of Nanjing Medical University, Changzhou, 213003, People’s Republic of China*These authors contributed equally to this workCorrespondence: Fang Su; Zishu Wang, Department of Medical Oncology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, 233004, People’s Republic of China, Tel +86 13605523272; +86 13955254185, Email sufang@bbmc.edu.cn; wzshahbb@163.comBackground: Research has revealed that Plexin domain containing 1 (PLXDC1) is correlated with the prognosis of a variety of tumors, but its role in the tumor microenvironment (TME) of gastric cancer has not been reported.Methods: In this study, we analyzed PLXDC1 expression in gastric cancer using the Oncomine and the Cancer Genome Atlas (TCGA) databases and immunohistochemical staining experiments, and performed prognostic assessment with data from the TCGA and Kaplan–Meier Plotter databases. The immunomodulatory role of PLXDC1 in the gastric cancer TME was analyzed by signaling pathway enrichment, immune cell correlation analysis, immunomodulator risk model construction and immunohistochemical staining experiments of immune cells.Results: The results indicated that PLXDC1 was overexpressed in gastric cancer and that its overexpression was associated with poor prognosis. Multivariate Cox analysis revealed that PLXDC1 could be an independent biomarker of the risk of gastric cancer. Signaling pathway enrichment revealed that high PLXDC1 expression was involved in signaling pathways related to immune activation and stromal activation, and Tumor Immune Dysfunction and Exclusion (TIDE) assessment indicated that high PLXDC1 expression was associated with a significantly higher risk of immune evasion than low PLXDC1 expression. A Cox risk model based on PLXDC1-associated immunomodulators also presented poor prognosis, and immune evasion was significantly higher in the high-risk group than in the low-risk group. In addition, immunohistochemical staining of CD8/CD3/CD4+ T cells in the high and low PLXDC1 expression groups also observed immune cell distribution characteristics of immune evasion.Conclusion: This study analyzed PLXDC1 from multiple biological perspectives and revealed that PLXDC1 can be a biomarker for poor prognosis and immune evasion in gastric cancer.Graphical Abstract: Keywords: PLXDC1, biomarker, tumor microenvironment, immune evasion, prognosis, immunotherapy

Ankyrin repeat and fibronectin type III domain containing 1 (ANKFN1) is reported to be involved in human height and developmental abnormalities, but the expression profile and molecular function of ANKFN1 in hepatocellular carcinoma (HCC) remain unknown. This study aimed to evaluate the clinical significance and biological function of ANKFN1 in HCC and investigate whether ANKFN1 can be used for differential diagnosis in HCC. Here, we showed that ANKFN1 was upregulated in 126 tumor tissues compared with adjacent nontumorous tissues in HCC patients. The upregulation of ANKFN1 in HCC was associated with cirrhosis, alpha-fetoprotein (AFP) levels and poor prognosis. Moreover, silencing ANKFN1 expression suppressed HCC cell proliferation, migration, invasion, and metastasis in vitro and subcutaneous tumorigenesis in vivo. However, ANKFN1 overexpression promoted HCC proliferation and metastasis in an orthotopic liver transplantation model and attenuated the above biological effects in HCC cells. ANKFN1 significantly affected HCC cell proliferation by inducing G1/S transition and cell apoptosis. Mechanistically, we demonstrated that ANKFN1 promoted cell proliferation, migration, and invasion via activation of the cyclin D1/Cdk4/Cdk6 pathway by stimulating the MEK1/2-ERK1/2 pathway. Moreover, ANKFN1-induced cell proliferation, migration, and invasion were partially reversed by ERK1/2 inhibitors. Taken together, our results indicate that ANKFN1 promotes HCC cell proliferation and metastasis by activating the MEK1/2-ERK1/2 signaling pathway. Our work also suggests that ANKFN1 is a potential therapeutic target for HCC.

In this study, experimental investigations were carried out on thin-ply and thick-ply countersunk interference fit joints. A comparative study of the mechanical behaviour and hole wall damage during static installation of thin-ply and thick-ply specimens with varied interference sizes were carried out. The effect of interference fit on the strength of countersunk composite bolted joints with different thicknesses of prepreg was investigated in detail under quasi-static tensile loading. The internal damage and failure mechanisms of thin-ply and thick-ply interference joints were investigated by SEM micrographs. The thin-ply specimens had lower installation forces and the initial damage of the thin-ply interference interface was less than that of the thick-ply for the same interference size. The thin-ply can still suppress the generation and propagation of the damage in the load-bearing of the interference connection. More importantly, thin-ply laminates can accommodate more substantial interference sizes than thick-ply laminates and have higher load-bearing strengths of approximately 10–20% more for the same interference size. Thin prepregs can be used to facilitate the design process and improve the structural properties of composites, providing new perspectives for improvements and innovative applications of interference joining technology.

The influenza virus, a viral respiratory pathogen, causes major morbidity and mortality worldwide. In this study, we compared and analyzed the effectiveness of the rescued two chimeric H1N1 vaccine strains rA/B-California/04/2009 and rA/B-California/07/2009, respectively, which expressed the hemagglutinin of A/California/07/2009 or A/California/04/2009 based on an influenza B virus master donor. The rA/B-California/07/2009 strain was rescued successfully. However, the rA/B-California/04/2009, which expressed the HA of A/California/07/2009 could hardly be generated. In substitution for alanine at residue 212 in the chimeric HA protein of rA/B-California/04/2009 virus, threonine was identified as a deleterious change. The free Gibbs energy change of mutating residue 212 reflected that threonine in site 212 caused a reduction in the stability of rA/B-California HAs. Additionally, with the analysis of binding free energy data and RMSD values of receptor analogs and two chimeric HAs, substitution 212T also conferred decreased receptor-binding avidity. These results suggested that the substitution A212T may enhance the stability and binding affinity of the chimeric H1N1 vaccine strains. This finding may help to design a live-attenuated influenza vaccine with more efficacy.

Influenza H3N8 viruses have been recovered frequently from wild bird species, including Anseriformes (primarily from migratory ducks) and Charadriiformes (primarily from shorebirds). However, little attention has been given to the transmission ability of H3N8 avian influenza viruses among mammals. Here, we study the potential human health threat and the molecular basis of mammalian transmissibility of H3N8 avian influenza viruses isolated from wild bird reservoirs. We classified eight H3N8 viruses into seven different genotypes based on genomic diversity. Six of eight H3N8 viruses isolated naturally from wild birds have acquired the ability to bind to the human-type receptor. However, the affinity for α-2,6-linked SAs was lower than that for α-2,3-linked SAs. Experiments on guinea pigs demonstrated that three viruses transmitted efficiently to direct-contact guinea pigs without prior adaptation. Notably, one virus transmitted efficiently via respiratory droplets in guinea pigs but not in ferrets. We further found that the PB1 S524G mutation conferred T222 virus airborne transmissibility between ferrets. We also determined that the 524G mutant increased viral pathogenicity slightly in mice compared with the WT (wild type). Based on these results, we elucidated the potential human health threat and molecular basis of mammalian transmissibility of H3N8 influenza viruses. We emphasized the need for continued surveillance of the H3N8 influenza viruses circulating in birds.

To explore the association of the levels of vascular endothelial growth factor (VEGF) and markers of epithelial − mesenchymal transition (EMT) in gastric cancer with histopathological features and long-term prognosis. Immunohistochemistry was used to quantitatively assess the expression of VEGF in gastric cancer tissues and adjacent tissues and to detect the expression level of EMT markers in gastric cancer tissues. Here we show that VEGF and EMT markers expression were significantly correlated with depth of tumor invasion and gastric cancer stage (P P P < 0.05), whereas VEGF and N-cadherin were positively correlated (r = 0.214, P < 0.05). Furthermore, Kaplan-Meier analysis and a Cox regression model were used to analyze the effect of VEGF and EMT marker expression on the survival of the patients. We found that the overall survival of gastric cancer patients was correlated with VEGF (P P P = 0.002) expression and parts of histopathological features. VEGF and EMT markers exist side by side and play a part together in the development of gastric cancer, which provides new ideas for evaluating the prognosis of gastric cancer and researching targeted drugs.

Background In 2011, a new influenza virus, named Influenza D Virus (IDV), was isolated from pigs, and then cattle, presenting influenza-like symptoms. IDV is one of the causative agents of Bovine Respiratory Disease (BRD), which causes high morbidity and mortality in feedlot cattle worldwide. To date, the molecular mechanisms of IDV pathogenicity are unknown. Recent IDV outbreaks in cattle, along with serological and genetic evidence of IDV infection in humans, have raised concerns regarding the zoonotic potential of this virus. Influenza virus polymerase is a determining factor of viral pathogenicity to mammals. Methods Here we take a prospective approach to this question by creating a random mutation library about PB2 subunit of the IDV viral polymerase to test which amino acid point mutations will increase viral polymerase activity, leading to increased pathogenicity of the virus. Results Our work shows some exact sites that could affect polymerase activities in influenza D viruses. For example, two single-site mutations, PB2-D533S and PB2-G603Y, can independently increase polymerase activity. The PB2-D533S mutation alone can increase the polymerase activity by 9.92 times, while the PB2-G603Y mutation increments the activity by 8.22 times. Conclusion Taken together, our findings provide important insight into IDV replication fitness mediated by the PB2 protein, increasing our understanding of IDV replication and pathogenicity and facilitating future studies.

Objectives: Civil servants image is one of the most important representatives of government image. Therefore, it is of great significance to study the factors affecting the public’s attitud...

The avian influenza virus H13 subtype circulates primarily in waterfowl. To explore the ability of the H13 virus to cross the host barriers, we genetically analysed two H13 isolates from wild birds in China and evaluated the infectivity of these subtypes in chickens. Genetic and molecular analyses showed differences in the lineages and amino acid sequences between the two subtypes; A/mallard/Dalian/DZ-137/2013 (H13N6) belonged to Group I, while A/Eurasian Curlew/Liaoning/ZH-385/2014 (H13N8) belonged to Group III. The nucleotide sequence results showed high homology (approximately 96.9%-100%) to sequences in GenBank. Neither H13 isolate replicated in adult chickens or 20-day-old chicks; however, the H13N8 strain replicated in 1- and 10-day-old chicks. Viruses were recovered from the nasal turbinate, tracheal, lung and colon tissues of chicks at 1, 3 and 5 days post-inoculation. The H13N6 isolates replicated inefficiently in 1-day-old chicks and did not replicate in 10-day-old chicks. Serological surveillance results showed that domestic chickens had a 4.6%-10.4% (15/328-34/328) positive antibody titre to the H13 virus. H13N6 and H13N8 isolates replicated in mammalian cell lines, including 293T, Madin-Darby canine kidney and chicken embryo fibroblast cells. Our results suggest that the AIV H13 subtype may cross the host barrier from wild waterfowl to land fowl.

Microchromosome maintenance protein 7 (MCM7), a DNA replication permitting factor, plays an essential role in initiating DNA replication. MCM7 is reported to be involved in tumor formation and progression, whereas the expression profile and molecular function of MCM7 in colorectal cancer (CRC) remain unknown. In this study, we aimed to evaluate the clinical significance and biological function of MCM7 in CRC and investigated whether MCM7 can be used for a differential diagnosis in CRC and whether it may serve as a more sensitive proliferation marker for CRC evaluation. Moreover, immunohistochemical analysis of MCM7 was performed in a total of 89 specimens, and high MCM7 expression levels were associated with worse overall survival (OS) in CRC patients. Furthermore, the cell functional test suggested that lentivirus-mediated silencing of MCM7 with shRNA in CRC cells significantly inhibited cellular proliferation and promoted apoptosis in vitro and inhibited tumor growth in vivo. Additionally, mechanistic studies further demonstrated that P21-activated protein kinase 2 (PAK2) was regulated by MCM7 via microarray analysis and cell functional recovery tests, and miR-107 played a role in regulating expression MCM7 via miRNA microarray analysis and 3’UTR reporter assays. Taken together, our results suggest that the miR-107/MCM7/PAK2 pathway may participate in cancer progression and that MCM7 may serve as a prognostic biomarker in CRC

Evidence on the impacts of traffic-related air pollution (TRAP) on ST-segment elevation myocardial infarction (STEMI) events is limited. We aimed to assess the acute effects of TRAP exposure on the clinical onset of STEMI and related cardiac impairments.We recruited patients who were admitted for STEMI and underwent primary percutaneous coronary intervention at Peking University Third Hospital between 2014 and 2020. Indicators relevant to cardiac impairments were measured. Concomitantly, hourly concentrations of traffic pollutants were monitored throughout the study period, including fine particulate matter, black carbon (BC), particles in size ranges of 5-560 nm, oxides of nitrogen (NOOur findings extend current understanding that short-term exposure to higher levels of traffic pollution was associated with increased STEMI risks and exacerbated cardiac impairments, and provide evidence on traffic pollution control priority for protecting vulnerable populations who are at greater risks of cardiovascular events.

H9N2 influenza virus has been reported worldwide for several decades, and it has evolved into multiple genotypes among domestic poultry. However, the study involving ecology and evolution of low pathogenic avian influenza virus H9N2 in wild birds in China is limited. Here, we carried out surveillance of avian influenza virus H9N2 in wild birds along with the East Asian-Australian migratory flyway in China in 2017. To estimate the prevalence of H9N2 avian virus in wild birds, information on exposure of wild bird populations to H9N2 viruses using serology, in addition to virology, would greatly improve monitoring capabilities. In this study, we also present serological data of H9N2 among wild birds in China during 2013-2016. We report the identification of poultry-derived H9N2 isolates from asymptomatic infected multi-species wild birds such as Common kestrel (Falco tinnunculus), Northern goshawk (Accipiter gentilis), Little owl (Athene noctua), and Ring-necked Pheasant (Phasianus colchicus) in North China in June 2017. Phylogenetic analysis demonstrated that Tianjin H9N2 isolates belong to the G81 and carry internal genes highly homologous to human H10N8 and H7N9. The isolates could directly infect mice without adaptation but were restricted to replicate in the respiratory system. Glycan binding preference analyses suggested that the H9N2 isolates have acquired a binding affinity for the human-like receptor. Notably, results from transimission experiment in guinea pigs and ferrets demonstrated the wild birds-derived H9N2 influenza virus exhibits efficient transmission phenotypes in mammalian models via respiratory droplets. Our results indicate that the H9N2 AIVs continued to circulate extensively in wild bird populations and migratory birds play an important role in the spread and genetic diversification of H9N2 AIVs. The pandemic potential of H9N2 viruses demonstrated by aerosol transmission in mammalian models via respiratory droplets highlights the importance of monitoring influenza viruses in these hosts.

Gastric cancer is a common malignancy in China, with the second highest mortality rate worldwide. Advanced gastric cancer usually exhibits a poor prognosis with a low 5-year survival rate. Therefore, developing novel drugs for the treatment of this cancer will be beneficial for patients. Demethylzeylasteral, an extract of tripterygium wilfordii, has shown positive anticancer activities. However, the possible antitumor effect of demethylzeylasteral on gastric cancer cells and its underlying molecular mechanism remain to be determined. In the present study, the Cell Counting Kit-8 and colony formation assays revealed that demethylzeylasteral impeded the proliferation of human gastric cancer cells in a dose-dependent manner. Furthermore, the Transwell assay identified an inhibitory effect of demethylzeylasteral on the migration of MKN-45 cells, while flow cytometry found that treatment with demethylzeylasteral induced apoptosis and decreased the mitochondrial membrane potential in the cancer cells. Further investigation revealed that demethylzeylasteral downregulated the phosphorylation of ERK1/2, AKT, and GSK-3β in MKN-45 cells. Notably, decreased expression of Bcl-2 and increased expression of Bax, cleaved caspase-3, cleaved caspase-9 and cleaved PARP were detected in the cancer cells treated with demethylzeylasteral. The present study demonstrated that demethylzeylasteral exhibits therapeutic potential for gastric cancer.

An organocatalytic [4 + 2] cascade annulation of salicylaldehydes and 1,3-bisarylsulfonylpropenes has been developed. This protocol enables the efficient and straightforward synthesis of a new series of 3-sulfonyl-2-sulfonylmethyl-2H-chromenes that are useful for exploring pharmacologically valued compounds. Further reductive modifications result in 3-desulfonylated chromene or chromane derivatives. This protocol can be expanded to the synthesis of 3-sulfonyl-2-sulfonylmethyl 1,2-dihydroquinoline.

Physical attractiveness can greatly influence business job applications (the "beauty premium" effect). However, little is known about whether and how physical attractiveness influences interviewers' evaluations of Chinese civil servant applicants, given that many characteristics of civil service appear to be different from those of business jobs. Using event-related potentials (ERPs), the current study investigated how female job candidates' physical attractiveness influenced interviewers' evaluations in Chinese civil servant interviews for both technical and managerial positions. The behavioral results showed that for the managerial positions, attractive female candidates had a much higher acceptance rate than unattractive candidates. However, for the technical positions, no significant difference was found between attractive and unattractive candidates. At the brain level, for the managerial positions, pairs of attractive faces with managerial posts elicited smaller N400 and larger late positive potential (LPP) amplitudes than did pairs of unattractive faces with managerial posts. However, this relationship was not observed for technical posts. The negative correlation between N400 amplitude and acceptance rate as well as the positive correlation between LPP amplitude and acceptance rate further confirmed these results. The present study suggests that beauty could potentially influence if candidates are accepted in real Chinese civil servant interviews, as observed experimentally in this research.

It has been documented that the purification of inclusion bodies from Escherichia coli by size exclusion chromatography (SEC) may benefit subsequent refolding and recovery of recombinant proteins. However, loading volume and the high cost of the column limits its application in large-scale manufacturing of biopharmaceutical proteins. We report a novel process using polyethylene glycol (PEG) precipitation under denaturing conditions to replace SEC for rapid purification of inclusion bodies containing recombinant therapeutic proteins. Using recombinant human interleukin 15 (rhIL-15) as an example, inclusion bodies of rhIL-15 were solubilized in 7 M guanidine hydrochloride, and rhIL-15 was precipitated by the addition of PEG 6000. A final concentration of 5% (w/v) PEG 6000 was found to be optimal to precipitate target proteins and enhance recovery and purity. Compared to the previously reported S-200 size exclusion purification method, PEG precipitation was easier to scale up and achieved the same protein yields and quality of the product. PEG precipitation also reduced manufacturing time by about 50 and 95% of material costs. After refolding and further purification, the rhIL-15 product was highly pure and demonstrated a comparable bioactivity with a rhIL-15 reference standard. Our studies demonstrated that PEG precipitation of inclusion bodies under denaturing conditions holds significant potential as a manufacturing process for biopharmaceuticals from E. coli protein expression systems.

Three-dimensional (3D) face reconstruction refers to the restoration and reconstruction of 3D model of face from one or more two-dimensional (2D) images. It has been widely used in face recognition, expression migration, face editing and other aspects. In the current existing algorithms, there are still many shortcomings in how to reconstruct 3D face by parametric model in real time. In this paper, based on the convolutional neural network, we integrate the weight mask into the loss function, and then use the back propagation algorithm to calculate the parameter gradient error. Finally, the parameter self-renewal purpose of the loss function is achieved by gradient descent. It can be seen from the experimental results that this method can accurately reconstruct the 3D contour of the face, and the reconstruction results are complete and the topological structure is known. This is very important for the application after face reconstruction, such as face changing, expression changing and other aspects of accuracy has been greatly improved.

Objective: To inspect the feasibility of recombinant stable HEK293 cell lines development for biopharmaceuticals production using CRISPR/Cas9-mediated site-specific integration. Results: Using EGFP as a model protein, we first confirmed that the ‘safe harbor’ AAVS1 locus could be successfully targeted and the exogenous genes could be integrated through homology-directed repair induced by CRISPR/Cas9 technology. Then we constructed a donor plasmid harboring CTLA4Ig gene with an upstream CMV promoter and a downstream puromycin N-acetyltransferase gene to accelerate the efficient integration and selection of CTLA4Ig expression clones. After puromycin enrichment, the transfected pool was diluted for single clone selection, and 12 recombinant clones with CTLA4Ig expression were finally selected with a targeting efficiency of 25.8%. Productivity assay demonstrated that a frequency of 83.3% of selected clone were of consistent productivities, thus illustrating the high efficiency and success rate of this strategy. Conclusions: CRISPR/Cas9 mediated site-specific integration is an efficient and reliable tool to establishment recombinant stable HEK293 cell lines for both academic and industrial applications.

Since augmented reality (AR) technology has the characteristics of enhancing display effect and real interaction with human beings, it has been widely used in different scenarios of education. However, the AR technology usually requires a corresponding equipment such as a head mounted display (HMD) to display, so its application scope is limited in many aspects. This paper uses the Unity development tools to develop an AR application, which can display three-dimensional (3D) structural models corresponding to different three views of a geometric object on mobile smartphones. During the learning process of engineering drawing, different three views can be scanned by the application, and the corresponding 3D model is automatically restored. By combining the virtual 3D structure with the real world experience, students can better learn engineering drawing courses in the virtual combined environment. The survey results show that the AR application can effectively improve students’ understanding and cognizing of 3D objects in engineering drawing learning.

With the rapid development of computer vision technology and the wide use of cameras, vision-based positioning has become a new research hot spot. Higher accuracy, lower cost, wider range of applications and some other advantages make vision-based positioning technology more promising than traditional positioning methods. This paper gives a briefly introduction of different methods of positioning firstly. Then we discuss different technologies of vision-based positioning techniques and their core algorithms. In addition, we compared the advantages and disadvantages of different algorithms. Finally, we investigate different applications of vision-based positioning and analyze the existing problems and possible development trends.

Aiming at the difficulty of detecting the depth of the internal defects in the longitudinal region of the heavy haul and high speed rail welds, this paper studies a technique of ultrasonic virtual source imaging for detecting internal defects in dual-layer media. The numerical simulation model of phased-array sub-aperture virtual source ultrasonic transmission and receiving in dual-layer medium was established by Snell's rule. The experimental verification was carried out to realize the accurate detection of internal defects of a series longitudinal distribution in the standard test block. The results show that the algorithm can effectively improve the detection rate of multiple defects in the longitudinal distribution of standard test block and significantly increase the signal-to-noise ratio.

Chinese hamster ovary (CHO) cells are the most widely used mammalian hosts for recombinant protein production. However, by conventional random integration strategy, development of a high-expressing and stable recombinant CHO cell line has always been a difficult task due to the heterogenic insertion and its caused requirement of multiple rounds of selection. Site-specific integration of transgenes into CHO hot spots is an ideal strategy to overcome these challenges since it can generate isogenic cell lines with consistent productivity and stability. In this study, we investigated three sites with potential high transcriptional activities: C12orf35, HPRT, and GRIK1, to determine the possible transcriptional hot spots in CHO cells, and further construct a reliable site-specific integration strategy to develop recombinant cell lines efficiently. Genes encoding representative proteins mCherry and anti-PD1 monoclonal antibody were targeted into these three loci respectively through CRISPR/Cas9 technology. Stable cell lines were generated successfully after a single round of selection. In comparison with a random integration control, all the targeted integration cell lines showed higher productivity, among which C12orf35 locus was the most advantageous in both productivity and cell line stability. Binding affinity and N-glycan analysis of the antibody revealed that all batches of product were of similar quality independent on integrated sites. Deep sequencing demonstrated that there was low level of off-target mutations caused by CRISPR/Cas9, but none of them contributed to the development process of transgene cell lines. Our results demonstrated the feasibility of C12orf35 as the target site for exogenous gene integration, and strongly suggested that C12orf35 targeted integration mediated by CRISPR/Cas9 is a reliable strategy for the rapid development of recombinant CHO cell lines.

In this paper, a multi-aperture synthetic focusing technique based on virtual source is studied, which improves the lateral resolution and depth of the ultrasonic imaging with respect to the single array dynamic synthetic aperture focusing technique (SAFT). It only needs to store the N numbers data for the A scan line, and the complexity of the algorithm is greatly reduced compared with the full focus technology (TFM) with the maintenance of image quality. Using Field II simulation and Multi2000 device experiment results show that compared with the single element dynamic synthetic aperture technology, virtual source multi-aperture synthetic focusing ultrasonic technique (VS-MSAFT) effectively improves the lateral resolution and depth of permeation, furthermore to improve the quality of ultrasound imaging.