The coronavirus disease 2019 (COVID-19) is still causing a wide range of infections and deaths due to the high variability of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Therefore, it is necessary to establish a reliable and convenient pseudovirus-based neutralization assay to develop drug targeted variants of SARS-CoV-2. Based on the HIV-1 backbone, we generated a high titer luciferase (Luc)-expressing pseudovirus packaging system. Three dominant S mutant substitution pseudovirus were also established and identified compared to wide type in hACE2-overexpressing HEK-293T cells (293T-ACE2 cells). Compared to serine protease inhibitor camostat mesylate, the cysteine protease inhibitor E-64d could significantly block all SARS-CoV-2 mutant S pseudovirus infection in 293T-ACE2 cells. Furthermore, the neutralization ability of two antibodies targeted receptor-binding domain (RBD) of SARS-CoV-2 spike protein (S) was evaluated, which showed different inhibition dose-effect curves among four types of S pseudovirus. Overall, we developed a pseudovirus-based neutralization assay for SARS-CoV-2, which would be readily adapted to SARS-CoV-2 variants for evaluating antibodies.

BTB and CNC homology1 (BACH1), working as a transcriptional factor, is demonstrated to function on the regulation of epigenetic modifications by complex regulatory networks. Although BACH1 is reported as an oncogene, the overall analysis of its role remains lacking. In this study, we uncovered the capacity of BACH1 as a new pan-cancer therapeutic target. We found that BACH1 is highly expressed in abundant cancers and correlated with the poor prognosis of most cancers. The mutation sites of BACH1 varied in different cancer types and correlated to patients' prognoses. The tumor mutation burden (TMB) in four cancer species and up to six tumor infiltrated immune cells had a significant relevance with BACH1. The enrichment analysis showed that the BACH1-associated genes were significantly enriched in the pathways of PD-1/L1 expression, ubiquitin-mediated proteolysis, T cell receptor, Th17 cell differentiation. We then demonstrated that BACH1 is positively correlated with the expression of many candidate genes, incluing SRPK2, GCLM, SLC40A1, and HK2 but negatively correlated with the expression of KEAP1 and GAPDH. Overall, our data shed light on BACH1's effect on latent utility in cancer targeting therapy.

O‐GalNAc glycosylation is the initial step of the mucin‐type O‐glycosylation. In humans, it is catalyzed by a family of 20 homologous UDP‐GalNAc:polypeptide N‐acetylgalactosaminyltransferases (ppGalNAc‐Ts). So far, there is very limited information on their protein substrate specificities. In this study, we developed an on‐chip ppGalNAc‐Ts assay that could rapidly and systematically identify the protein substrates of each ppGalNAc‐T. In detail, we utilized a human proteome microarray as the protein substrates and UDP‐GalNAz as the nucleotide sugar donor for click chemistry detection. From a total of 16 368 human proteins, we identified 570 potential substrates of ppGalNAc‐T1, T2, and T3. Among them, 128 substrates were overlapped, while the rest were isoform specific. Further cluster analysis of these substrates showed that the substrates of ppGalNAc‐T1 had a closer phylogenetic relationship with that of ppGalNAc‐T3 compared with ppGalNAc‐T2, which was consistent with the topology of the phylogenetic tree of these ppGalNAc‐Ts. Taken together, our microarray‐based enzymatic assay comprehensively reveals the substrate profile of the ppGalNAc‐T1, T2, and T3, which not only provides a plausible explanation for their partial functional redundancy as reported, but clearly implies some specialized roles of each enzyme in different biological processes.

The effect of adjuvant radiotherapy on the survival outcomes of patients with mucinous rectal cancer remains unclear. This study evaluated the 5-year cause specific survival (CSS) of patients with mucinous rectal cancer after surgery to determine whether adjuvant radiotherapy conferred a survival benefit.

BACKGROUND:Glycolysis is considered to be the root of cancer development and progression, which involved a multi-step enzymatic reaction. Our study aimed at figuring out which glycolysis enzyme participates in the development of colorectal cancer and its possible mechanisms.METHODS:We firstly screened out Aldolase B (ALDOB) by performing qRT-PCR arrays of glycolysis-related genes in five paired liver metastasis and primary colorectal tissues, and further detected ALDOB protein with immunohistochemistry in tissue microarray (TMA) consisting of 229 samples from stage I-III colorectal cancer patients. CRISPR-Cas9 method was adopted to create knock out colon cancer cell lines (LoVo and SW480) of ALDOB. The effect of ALDOB on cell proliferation and metastasis was examined in vitro using colony formation assay as well as transwell migration and invasion assay, respectively.RESULTS:In TMA, there was 64.6% of samples demonstrated strong intensity of ALDOB. High ALDOB expression were associated with poor overall survival and disease-free survival in both univariate and multivariate regression analyses (P<0.05). In vitro functional studies of CCK-8 demonstrated that silencing ALDOB expression significantly (P<0.05) inhibited proliferation, migration and invasion of colon cancer cells. Mechanically, silencing ALDOB activated epithelial markers and repressed mesenchymal markers, indicating inactivation of ALDOB may lead to inhibition of epithelial-mesenchymal transition (EMT).CONCLUSION:Upregulation of ALDOB promotes colorectal cancer metastasis by facilitating EMT and acts as a potential prognostic factor and therapeutic target in colorectal cancer.

An efficient method for construction of 3-arylquinolin-4(1H)-ones via in situ Meinwald rearrangement/intramolecular reductive cyclization of 2′-nitrochalcone epoxides has been developed. The practical approach is of excellent functional groups compatibility with as high as 98% yield under mild reaction conditions. Trapping and NMR analysis about the key intermediates of the transformation provided insights to propose a plausible mechanism for the intramolecular reductive cyclization. Moreover, further derivation successfully furnished hydroxyl substituted and N-methyl substituted derivatives which may provide a promising potential application in exploring biologically active compounds of 3-arylquinolin-4(1H)-ones.

Semiconductor devices like transistors, diodes and ICs have some characteristic electrical noise property. At low frequency, this is of a "1/f" type. Devices with defects in the surface region show more noise signal than the normally good devices. This has been observed in the experiments conducted using the Quan-Tech Noise Analyzer. The graphs clearly indicate the character of more noisy components. Noise measurement can be used as effective screening method for semiconductor devices. The present paper deals with the noise analysis of Indian manufactured samples of transistors, zeners and linear integrated circuits. The screening technique is applicable to all semiconductor devices. The study on the correlation of external stress to produce the surface defect, the noise level and subsequent failure mode has been proposed. Noise analysis screening method helps in rejecting the more noisy devices, which are prone to early failures during the instrument's life. This way it helps improve the reliability of electronic instruments.

Effective drug delivery to a tumor depends on favorable blood perfusion within the tumor. As an important component of tumor extracellular matrix, fibrin is abundant near tumor vessels. Inspired by the distinct distribution pattern and vessel-dependent production of fibrin, we hypothesized that fibrin depletion in tumors decompresses tumor vessels to improve tumor blood perfusion and accordingly enhance drug delivery to tumors rich in vessels. In the present study, we attempted to employ a clinically used thrombolytic drug, recombinant tissue plasminogen activator (rtPA), to modulate fibrin deposition in tumors. We then combined this drug with a nanoparticle drug delivery system for tumor therapy. RtPA treatment (25 mg/kg/d i. p. administration for two weeks) successfully depleted fibrin deposition and enhanced blood perfusion within A549 tumor xenografts. Furthermore, rtPA treatment also improved the in vivo delivery of 115-nm nanoparticles to tumor tissues. Finally, rtPA combined with therapeutic agent-loaded nanoparticles resulted in the most effective shrinkage of A549 tumor xenografts compared with the control groups. Overall, the present study provides a new strategy to enhance the delivery of nanotherapeutics to tumors rich in vessels.

In this study, the photothermal effect of magnetic nanoparticle clusters was firstly reported for the photothermal ablation of tumors both in vitro in cellular systems but also in vivo study. Compared with individual magnetic Fe3O4 nanoparticles (NPs), clustered Fe3O4 NPs can result in a significant increase in the near-infrared (NIR) absorption. Upon NIR irradiation at 808 nm, clustered Fe3O4 NPs inducing higher temperature were more cytotoxic against A549 cells than individual Fe3O4 NPs. We then performed in vivo photothermal therapy (PTT) studies and observed a promising tumor treatment. Compared with PBS and individual magnetic Fe3O4 NPs by NIR irradiation, the clustered Fe3O4 NPs treatment showed a higher therapeutic efficacy. The treatment effects of clustered Fe3O4 NPs with different time of NIR illumination were also evaluated. The result indicated that a sustained high temperature generated by NIR laser with long irradiation time was more effective in killing tumor cells. Furthermore, histological analysis of H&E staining and TUNEL immunohistological assay were further employed for antitumor efficacy assessment of PTT against A549 tumors.

Although regional lymph nodes (RLN) dissection remains the only way to cure pancreatic cancer metastasis, it is unavoidably associated with sizable trauma, multiple complications, and low surgical resection rates. Thus, exploring a treatment approach for the ablation of drug-resistant pancreatic cancer is always of great concern. Moreover, reoperative and intraoperative mapping of RLN is also important during treatment, because only a few lymph nodes can be detected by the naked eye. In our study, graphene oxides modified with iron oxide nanoparticles (GO-IONP) as a nanotheranostic agent is firstly developed to diagnose and treat RLN metastasis of pancreatic cancer. The approach was designed based on clinical practice, the GO-IONP agent directly injected into the tumor was transported to RLN via lymphatic vessels. Compared to commercial carbon nanoparticles currently used in the clinic operation, the GO-IONP showed powerful ability of dual-modality mapping of regional lymphatic system by magnetic resonance imaging (MRI), as well as dark color of the agent providing valuable information that was instrumental for surgeon in making the preoperative plan before operation and intraoperatively distinguish RLN from surrounding tissue. Under the guidance of dual-modality mapping, we further demonstrated that metastatic lymph nodes including abdominal nodes could be effectively ablated by near-infrared (NIR) irradiation with an incision operation. The lower systematic toxicity of GO-IONP and satisfying safety of photothermal therapy (PTT) to neighbor tissues have also been clearly illustrated in our animal experiments. Using GO-IONP as a nanotheranostic agent presents an approach for mapping and photothermal ablation of RLN, the later may serve as an alternative to lymph node dissection by invasive surgery.

This study developed a simple and rapid purification method for plasma membrane with high yields from adherent cells. The plasma membrane (PM) sheets could be absorbed specifically by the cationic silica-magnetite nanocomposites (CSMN) under acidic conditions, and recovered directly in cell-lysis-buffer with no need for precipitation. The binding between CSMN and PM sheets was confirmed by electron microscopy. Western blot analysis demonstrated a 410-fold relative enrichment factor. Up to 422 integral membrane proteins were identified from 10(7) Huh7 cells. Notably, we found 29 Ras family proteins by classification according to their biological functions. The whole enrichment procedure took <30 min. The CSMN-based procedure demonstrates a simple, economical and efficient enrichment of integral PM proteins in proteomic study.

Hemodynamic characteristics such as blood velocity,blood pressure,flow impingement,wall shear stress and oscillatory shear index are considered to play important roles in the initiation,growth,rupture and recurrence of the cerebral aneurysms.Endovascular therapy is widely implemented to treat the cerebral aneurysms by releasing coils into the aneurysm sac for limiting the blood flow to the sac and stent-assisted coil embolization is adopted to occlude the wide-necked or complex aneurysms.Some researchers believe that stents are not only a mechanical device but may act as a biological system and contribute to vessel wall healing.Hemodynamics simulation helps people understand the effect of hemodynamic characteristics on the recurrence of the coiled aneurysm and it also benefits the interventional planning of neurosurgeons.This study constructed the numerical model for a subject-specific ICA aneurysm treated with stent-assisted coil embolization,which combined the coiled model of the aneurysm with a porous stent placement,and simulated the pulsatile blood flow in these aneurysm models.When a stent was placed across the aneurysm orifice in the coiled aneurysm,the high wall shear stress around the distal aneurysm root was reduced more than that of the coiled aneurysm without a stent.The simulated results point to the conclusion that the stent not only protects the parent artery from occlusion due to extension of coils or thrombosis,but may also reduce the recurrence risk of the stent-assisted coiled aneurysm.

O-glycosylation is an important post translational modification of proteins. It is the main type of protein glycosylation as well as N-glycosylation. Because of its significant roles in modulating the function and structure of proteins, it is necessary to study protein O-glycosylation in biomedicine research. The main tasks of protein O-glycosylation analysis are; (1) identification of O-glycosylated proteins, (2) location of O-glycosylation site on proteins, (3) O-glycan structure interpretation and (4) quantitative analysis. Mass spectrometry (MS) has become the key technology for the analysis of protein O-glycosylation. However, the analysis of O-glycosylation is a challenge; there is no consensus sequence of O-glycosylation and no universal cleavage enzyme, and the O-glycans are very complicated. Besides the invention of new instruments like LTQ Velos, many novel methods have been reported recently to conquer this challenge. In this review we try to cover these improvements for analysis of protein O-glycosylation, including four aspects; enrichment of O-glycoproteins/peptides, dissociation of O-glycan from proteins, O-glycan structure identification and quantitative analysis.

Alternating current (AC) has been employed to enhance the efficiency of chymotryptic proteolysis for peptide mapping. It was allowed to flow through the mixture solution of proteins and chymotrypsin via a pair of platinum wire electrodes. Bovine serum albumin (BSA) and cytochrome c (Cyt-c) were digested by the novel proteolysis approach to demonstrate its feasibility and performance. The results indicated that AC significantly accelerated in-solution chymotryptic proteolysis and the digestion time was substantially reduced to 5 min. The digests were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with sequence coverages of 46% (BSA) and 90% (Cyt-c) that were much better than those obtained by using 12-h conventional in-solution chymotryptic proteolysis. In addition, AC-assisted chymotryptic proteolysis was employed to digest human serum to demonstrate its suitability to complex protein sample. The present proteolysis strategy is simple and efficient and will find a wide range of applications in proteomic research.

In this report, trypsin was immobilized on silica-coated fiberglass core in microchip to form a core-changeable bioreactor for highly efficient proteolysis. To prepare the fiber core, a layer of organic–inorganic hybrid silica coating was prepared on the surface of a piece of glass fiber by a sol–gel method with tetraethoxysilane (TEOS) and 3-aminopropyltriethoxysilane (APTES) as precursors. Subsequently, trypsin was immobilized on the coating with the aid of glutaraldehyde. Prior to use, the enzyme-immobilized fiber was inserted into the channel of a microchip to form an in-channel fiber bioreactor. The novel bioreactor can be regenerated by changing its fiber core. The scanning electron microscopy images of the cross-section of a trypsin-immobilized fiber indicated that a layer of ∼1μm thick film formed on the glass substrate. The feasibility and performance of the unique bioreactor were demonstrated by the tryptic digestion of bovine serum albumin (BSA) and cytochrome c (Cyt-c) and the digestion time was significantly reduced to less than 10s. The digests were identified by MALDI-TOF MS with sequence coverages of 45% (BSA) and 77% (Cyt-c) that were comparable to those obtained by 12-h conventional in-solution tryptic digestion. The fiber-based microchip bioreactor provides a promising platform for the high-throughput protein identification.

Protein glycosylation as an important post-translational modification has a significant effect on the structures and functions of proteins. As a section of proteomics, glycoproteomics is in the spotlight currently, and the efficient separation and enrichment of glycoproteins/glycopeptides from complex biological samples is the key and difficult point of glycoproteome research. In this paper, the progress in methods for the separation and enrichment of glycoproteins/glycopeptides and their applications are overviewed. These methodologies not only include most-often-used methods such as lectin-based affinity chromatography, boronic acid method, hydrazide chemistry and hydrophilic chromatography, but also some novel methods, for example, size exclusion chromatography and strong cation exchange enrichment.

In this report, infrared (IR)-assisted on-plate proteolysis has been developed for rapid peptide mapping. Protein solutions containing trypsin were allowed to digest directly on the spots of matrix-assisted laser desorption/ionization (MALDI) plates under IR radiation. The feasibility and performance of the novel proteolysis approach were investigated by the digestion of bovine serum albumin (BSA) and cytochrome c (Cyt-c). It was demonstrated that IR radiation substantially enhanced the efficiency of proteolysis and the digestion time was significantly reduced to 5 min. The digests were identified by MALDI time-of-flight mass spectrometry with sequence coverages of 55 (BSA) and 75% (Cyt-c) that were comparable to those obtained by using conventional in-solution tryptic digestion. The suitability of IR-assisted on-plate proteolysis to complex proteins was demonstrated by digesting human serum and casein extracted from commercially available milk sample. The present proteolysis strategy is simple and efficient, offering great promise for high-throughput protein identification.

In this report, infrared (IR) radiation was employed to enhance the efficiency of tryptic proteolysis for peptide mapping. Protein solutions containing trypsin in sealed transparent Eppendorf tubes were allowed to digest under an IR lamp at 37 degrees C. The feasibility and performance of the novel proteolysis approach were demonstrated by the digestion of BSA and myoglobin (MYO) and the digestion time was significantly reduced to 5 min. The obtained digests were identified by MALDI-TOF MS with the sequence coverages of 69% (BSA) and 90% (MYO) that were much better than those obtained by conventional in-solution tryptic digestion. The present IR-assisted proteolysis strategy is simple and efficient, offering great promise for high-throughput protein identification.