Three-dimensional breast cancer tumor models based on natural hydrogels:a review

Breast cancer is the most common cancer in women and one of the deadliest cancers worldwide.According to the distribution of tumor tissue,breast cancer can be divided into invasive and non-invasive forms.The cancer cells in invasive breast cancer pass through the breast and through the immune system or systemic circulation to different parts of the body,forming metastatic breast cancer.Drug resistance and distant metastasis are the main causes of death from breast cancer.Research on breast cancer has attracted extensive attention from researchers.In vitro construction of tumor models by tissue engineering methods is a common tool for studying cancer mechanisms and anticancer drug screening.The tumor microenvironment consists of cancer cells and various types of stromal cells,including fibroblasts,endothelial cells,mesenchymal cells,and immune cells embedded in the extracellular matrix.The extracellular matrix contains fibrin proteins(such as types Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅵ,and Ⅹ collagen and elastin)and glycoproteins(such as proteoglycan,laminin,and fibronectin),which are involved in cell signaling and binding of growth factors.The current traditional two-dimensional(2D)tumor models are limited by the growth environment and often cannot accurately reproduce the heterogeneity and complexity of tumor tissues in vivo.Therefore,in recent years,research on three-dimensional(3D)tumor models has gradually increased,especially 3D bioprinting models with high precision and repeatability.Compared with a 2D model,the 3D environment can better simulate the complex extracellular matrix components and structures in the tumor microenvironment.Three-dimensional models are often used as a bridge between 2D cellular level experiments and animal experiments.Acellular matrix,gelatin,sodium alginate,and other natural materials are widely used in the construction of tumor models because of their excellent biocompatibility and non-immune rejection.Here,we review various natural scaffold materials and construction methods involved in 3D tissue-engineered tumor models,as a reference for research in the field of breast cancer.

Inflammatory granuloma of the trachea: a rare case with Epstin-Barr virus infection

Epstein-Barr virus(EBV),a double-stranded DNA virus with an envelope,is a ubiquitous pathogen that is prevalent in humans,although most people who contract it do not develop symptoms(Kerr,2019).While the primary cells EBV attacks are epithelial cells and B lymphocytes,its target range expands to a variety of cell types in immunodeficient hosts.Serological change occurs in 90%of infected patients.Therefore,immunoglobulin M(IgM)and IgG,serologically reactive to viral capsid antigens,are reliable biomarkers for the detection of acute and chronic EBV infections(Cohen,2000).Symptoms of EBV infection vary according to age and immune status.

High expression of FABP4 in colorectal cancer and its clinical significance

Objective:To investigate the relationship between the fatty acid-binding protein 4(FABP4)and colorectal cancer(CRC).Methods:Using an enzyme-linked immunosorbent assay(ELISA),we measured the expression of FABP4 in plasma of50 patients who underwent surgery for CRC from October 2017 to May 2018 and 50 healthy controls.The content of the visceral fat area(VFA)as seen with abdominal computed tomography(CT)scanning was measured by ImageJ software.The expression levels of FABP4,E-cadherin,and Snail proteins in CRC and adjacent tissues were determined by immunohistochemistry.Results:The mean concentration of plasma FABP4 of CRC patients was higher than that of the control group(22.46 vs.9.82 ng/m L;P<0.05).The concentration of plasma FABP4 was related to the tumor,node,metastatis(TNM)stage and lymph node metastasis and was independent of age,body mass index(BMI),tumor size and location,and the degree of differentiation of CRC.The concentration of plasma FABP4 was positively correlated with high VFA and lipoprotein-a(LPA)(P<0.05);but it was not correlated with total cholesterol(TG),total triglyceride(TC),low-density lipoprotein(LDL),high-density lipoprotein(HDL),or apolipoprotein AI(Apo-AI).The expression of FABP4 protein in CRC tissues was positively correlated with the degree of CRC differentiation,tumor stage,and lymph node metastasis.The level of FABP4 protein was negatively correlated with E-cadherin protein(r=-0.3292,P=0.0196)and positively correlated with Snail protein(r=0.5856,P<0.0001).Conclusions:High LPA and VFA were risk factors for increased plasma FABP4 in CRC patients.FABP4 protein was highly expressed in CRC tissues and associated with TNM stage,differentiation,and lymph node metastasis of CRC.The level of FABP4 in CRC tissue was correlated with E-cadherin and Snail expression,suggesting that FABP4 may promote CRC progression related to epithelial-mesenchymal transition(EMT).

Gastrointestinal toxicities associated with immune checkpoint inhibitors: a disproportionality analysis leveraging VigiBase,the WHO Adverse Drug Reaction Database

With the improvement of people's living standards, gastrointestinal adverse reactions caused by various adverse factors have attracted more and more people's attention. A recent study has indicated that coronavirus disease 2019(COVID-19) could also invade the gastrointestinal tract, leading to gastrointestinal adverse reactions(Song et al., 2020). In recent years, immunotherapy has provided certain effects for some patients with advanced malignant tumors.

Psychophysiological and cognitive effects of strawberry plants on people in isolated environments

In manned deep-space exploration,extremely isolated environments may adversely affect the mood and cognition of astronauts.Horticultural plants and activities have been proven to be effective in improving their physical,psychological,and cognitive states.To assess the effects of applying horticultural plants and activities in isolated environments,this study investigated the influence of viewing strawberry plants on the mood of people in a laboratory experiment as indicated by heart rate,salivary cortisol,and psychological scales.The results showed that heart rate and salivary cortisol were significantly decreased after viewing strawberry plants for 15 min.“Tension”and“confusion”scored using the Profile of Mood States negative mood subscales,and anxiety levels measured using the State-Trait Anxiety Inventory scale were also significantly reduced.This study further explored the impact of viewing strawberry plants on cognition.A notable reduction of the subjects’reaction time after 15-min plant viewing was observed.Based on these findings,a long-duration isolated experiment in a bioregenerative life support system—“Lunar Palace I”—was conducted.A similar trend was obtained that crew members’mood states were improved by viewing the strawberry plants,but no significant change was observed.This study provided some experimental evidence for the benefits of interacting with strawberry plants in isolated environments.

Central visual function and inner retinal structure in primary open-angle glaucoma

To investigate associations between central visual function and inner retinal structure in primary open-angle glaucoma(POAG). This study enrolled 78 POAG patients and 58 healthy controls. POAG was classified into early glaucoma and moderate to advanced glaucoma. The following tests were performed on all participants: isolated-check visual evoked potential(ic VEP) testing, 24-2 standard automated perimetry(SAP), and Cirrus optical coherence tomography(OCT) examinations. Signal-to-noise ratio(SNR) measures obtained from ic VEP responses to isolated checks presented at four depths of modulation(DOMs;8%, 14%, 22%, and 32%) were explored. Mean macular sensitivity(mMS) was assessed by calculating the mean sensitivities of central 12 SAP points. Ganglion cell layer+ inner plexiform layer thickness(GCL+IPLT) and peripapillary retinal nerve fiber layer thickness(pRNFLT) were measured by OCT scanning. For each group of subjects, linear relationships among the following measures were analyzed: SNR, mMS, GCL+IPLT, and pRNFLT. SNR, mMS, GCL+IPLT, and pRNFLT were all more significantly decreased in glaucoma than in controls(P<0.001). A significant positive association was found between SNR at 14% DOM and GCL+IPLT at the inferior sector in early glaucoma(r=0.465, P=0.004). In moderate to advanced glaucoma, significant correlations were found between SNR at 32% DOM and mean GCL+IPLT(r=0.364, P=0.023), superior GCL+IPLT(r=0.358, P=0.025), and mean p RNFLT(r=0.396, P=0.025). In addition, in moderate to advanced glaucoma, there were significant correlations between mMS and all relevant measures of retinal thickness(r=0.330–0.663, P< 0.010). In early glaucoma, significant correlations were found between mean mMS and minimum GCL+IPLT(r=0.373, P=0.023), and between inferior mMS and superior GCL+IPLT(r=0.470, P=0.003). Linear models provided a good explanation for the relationship between SNR and inner retinal thickness(IRT), whereas nonlinear models better explained the relationship between mMS and IRT. In early glaucoma, both SNR and mMS were related moderately and significantly to IRT, whereas in moderate to advanced glaucoma, mMS was more strongly correlated with IRT than SNR.

Regulation of flowering time via miR172-mediated APETALA2-like expression in ornamental gloxinia(Sinningia speciosa)

We investigated the microRNA172(miR172)-mediated regulatory network for the perception of changes in external and endogenous signals to identify a universally applicable floral regulation system in ornamental plants, manipulation of which could be economically beneficial. Transgenic gloxinia plants, in which miR172 was either overexpressed or suppressed, were generated using Agrobacterium-mediated transformation. They were used to study the effect of altering the expression of this miRNA on time of flowering and to identify its mRNA target. Early or late flowering was observed in transgenic plants in which miR172 was overexpressed or suppressed, respectively. A full-length complementary DNA(cDNA) of gloxinia(Sinningia speciosa) APETALA2-like(SsAP2-like) was identified as a target of miR172. The altered expression levels of miR172 caused up-or down-regulation of SsAP2-like during flower development, which affected the time of flowering. Quantitative real-time reverse transcription PCR analysis of different gloxinia tissues revealed that the accumulation of SsAP2-like was negatively correlated with the expression of miR172 a, whereas the expression pattern of miR172 a was negatively correlated with that of miR156 a. Our results suggest that transgenic manipulation of miR172 could be used as a universal strategy for regulating time of flowering in ornamental plants.

Bone morphogenetic proteins and inner ear development

Bone morphogenetic proteins(BMPs)are the largest subfamily of the transforming growth factor-βsuperfamily,and they play important roles in the development of numerous organs,including the inner ear.The inner ear is a relatively small organ but has a highly complex structure and is involved in both hearing and balance.Here,we discuss BMPs and BMP signaling pathways and then focus on the role of BMP signal pathway regulation in the development of the inner ear and the implications this has for the treatment of human hearing loss and balance dysfunction.

Depolymerized konjac glucomannan: preparation and application in health care

Konjac glucomannan（KGM） is a water-soluble polysaccharide obtained from the roots and tubers of konjac plants. Recently, a degraded product of KGM, depolymerized KGM（DKGM）, has attracted attention because of its low viscosity, improved hydrophily, and favorable physiological functions. In this review, we describe the preparation of DKGM and its prebiotic effects. Other health benefits of DKGM, covering antioxidant and immune activity, are also discussed, as well as its safety. DKGM could be a candidate for use as a tool for the treatment of various diseases, including intestinal flora imbalance, and oxidative-and immune-related disorders.

Menaquinone-7 production from maize meal hydrolysate by Bacillus isolates with diphenylamine and analogue resistance

A menaquinone-7（MK-7） high-producing strain needs to be isolated to increase MK-7 production, in order to meet a requirement of MK-7 given the low MK-7 content in food products. This article focuses on developing MK-7 high-producing strains via screening and mutagenesis by an atmospheric and room temperature plasma（ARTP） mutation breeding system. We isolated an MK-7-producing strain Y-2 and identified it as Bacillus amyloliquefaciens, which produced（7.1±0.5） mg/L of MK-7 with maize meal hydrolysate as carbon source. Then, an MK-7 highproducing strain B. amyloliquefaciens H.β.D.R.-5 with resistance to 1-hydroxy-2-naphthoic acid, β-2-thienylalanine, and diphenylamine was obtained from the mutation of the strain Y-2 using an ARTP mutation breeding system. Using strain H.β.D.R.-5, efficient production of MK-7 was achieved（（30.2±2.7） mg/L）. In addition, the effects of nitrogen sources, prenyl alcohols, and MgSO_4 on MK-7 production were investigated, suggesting that soymeal extract combined with yeast extract, isopentenol, and MgSO4 was beneficial. Under the optimized condition, the MK-7 production and biomass-specific yield reached（61.3±5.2） mg/L and 2.59 mg/L per OD600 unit respectively in a 7-L fermenter. These results demonstrated that strain H.β.D.R.-5 has the capacity to produce MK-7 from maize meal hydrolysate, which could reduce the substrate cost.

Molecular cloning, characterization, and promoter analysis of the isochorismate synthase （AalCS1） gene from Artemisia annua

Isochorismate synthase(ICS) is a crucial enzyme in the salicylic acid(SA) synthesis pathway. The full-length complementary DNA(cDNA) sequence of the ICS gene was isolated from Artemisia annua L. The gene, named AaICS1, contained a 1710-bp open reading frame, which encoded a protein with 570 amino acids. Bioinformatics and comparative study revealed that the polypeptide protein of AaICS1 had high homology with ICSs from other plant species. Southern blot analysis suggested that AaICS1 might be a single-copy gene. Analysis of the 1470-bp promoter of AaICS1 identified distinct cis-acting regulatory elements, including TC-rich repeats, MYB binding site(MBS), and TCA-elements. An analysis of AaICS1 transcript levels in multifarious tissues of A. annua using quantitative real-time polymerase chain reaction(qRT-PCR) showed that old leaves had the highest transcription levels. AaICS1 was up-regulated under wounding, drought, salinity, and SA treatments. This was corroborated by the presence of the predicted cis-acting elements in the promoter region of AaICS1. Overexpressing transgenic plants and RNA interference transgenic lines of AaICS1 were generated and their expression was compared. High-performance liquid chromatography(HPLC) results from leaf tissue of transgenic A. annua showed an increase in artemisinin content in the overexpressing plants. These results confirm that AaICS1 is involved in the isochorismate pathway.

Identification of neuron-related genes for cell therapy of neurological disorders by network analysis

Bone mesenchymal stem cells（BMSCs） differentiated into neurons have been widely proposed for use in cell therapy of many neurological disorders. It is therefore important to understand the molecular mechanisms underlying this differentiation. We screened differentially expressed genes between immature neural tissues and untreated BMSCs to identify the genes responsible for neuronal differentiation from BMSCs. GSE68243 gene microarray data of rat BMSCs and GSE18860 gene microarray data of rat neurons were received from the Gene Expression Omnibus database. Transcriptome Analysis Console software showed that 1248 genes were up-regulated and 1273 were down-regulated in neurons compared with BMSCs. Gene Ontology functional enrichment, protein-protein interaction networks, functional modules, and hub genes were analyzed using DAVID, STRING 10, BiN GO tool, and Network Analyzer software, revealing that nine hub genes, Nrcam, Sema3 a, Mapk8, Dlg4, Slit1, Creb1, Ntrk2, Cntn2, and Pax6, may play a pivotal role in neuronal differentiation from BMSCs. Seven genes, Dcx, Nrcam, Sema3 a, Cntn2, Slit1, Ephb1, and Pax6, were shown to be hub nodes within the neuronal development network, while six genes, Fgf2, Tgfβ1, Vegfa, Serpine1, Il6, and Stat1, appeared to play an important role in suppressing neuronal differentiation. However, additional studies are required to confirm these results.

Evaluation of potential anti-toxoplasmosis efficiency of combined traditional herbs in a mouse model

Toxoplasma gondii is a worldwide spread protozoan and is able to infect almost all warm-blood animals. No effective drugs are available clinically on toxoplasmosis. Chinese traditional herbal medicines have provided remedies for many health problems. There exists a possibility that Chinese herbs may provide protection against T. gondii. This work aims to assess the protective efficacy of combined Chinese herbs against T. gondii. We screened five herbal medicines that have different pharmacological effects and combined them into a prescription according to the traditional Chinese medicine compatibility principle. The drug potential and protective efficacy were evaluated through a mouse model by determining the survival time, the parasite load in blood and tissues, the change of cell proportions in blood and histological detection. The results showed that the survival time of mice in the 500 mg Chinese herbs group and sulfadiazine group was significantly longer than that of the PBS control group. Also the parasite load in blood and tissues of 500 mg Chinese herbs and sulfadiazine groups was significantly lower than that of PBS group at 7 days post infection（dpi）, which was in accordance with the result of histological detection. Monocyte and neutrophil of infected mice were remarkably increased while lymphocyte was dramatically decreased compared to that of blank group at 7 dpi. The results demonstrated that the 500 mg dosage of our Chinese herbs could slow down the replication of T. gondii and prolong the survival time of mice and could be considered as possible candidate drug against toxoplasmosis.

Genome-wide profiling of genetic variation in Agrobacterium-transformed rice plants

Agrobacterium-mediated transformation has been widely used in producing transgenic plants, and was recently used to generate ＂transgene-clean＂ targeted genomic modifications coupled with the clustered regularly interspaced short palindromic repeats （CRISPR）/CRISPR-associated （Cas9） system.

Cholangiocarcinoma-derived exosomes inhibit the antitumor activity of cytokine-induced killer cells by down-regulating the secretion of tumor necrosis factor-α and perforin

Objective： The aim of our study is to observe the impact of cholangiocarcinoma-derived exosomes on the antitumor activities of cytokine-induced killer （CIK） cells and then demonstrate the appropriate mechanism. Methods： Tumor-derived exosomes （TEXs）, which are derived from RBE cells （human cholangiocarcinoma line）, were collected by ultracentrifugation. CIK cells induced from peripheral blood were stimulated by TEXs. Fluorescence-activated cell sorting （FACS） was performed to determine the phenotypes of TEX-CIK and N-CIK （normal CIK） cells. The concen- trations of tumor necrosis factor-a （TNF-a） and perforin in the culture medium supematant were examined by using an enzyme-linked immunosorbent assay （ELISA） kit. A CCK-8 kit was used to evaluate the cytotoxic activity of the CIK cells to the RBE cell line. Results： The concentrations of TNF-a and perforin of the group TEX-CIK were 138.61 pg/ml and 2.41 ng/ml, respectively, lower than those of the group N-CIK 194.08 pg/ml （P〈0.01） and 3.39 ng/ml （P〈0.05）. The killing rate of the group TEX-CIK was 33.35%, lower than that of the group N-CIK （47,35% （P〈0.01））. The population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells decreased in the TEX-CIK group （63.2±6.8）%, （2.5±1.0）%, （0.53±0.49）%, （0.45±0.42）%） compared with the N-CIK group （（90.3±7.3）%, （65.7±3.3）%, （4.2±1.2）%, （15.2±2.7）%）, P〈0.01. Conclusions： Our results suggest that RBE cells-derived exosomes inhibit the antitumor activity of CIK cells by down-regulating the population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells and the secretion of TNF-a and perforin. TEX may play an important role in cholangiocarcinoma immune escape.

Molecular cloning, characterization and expression of the energy homeostasis-associated gene in piglet

The energy homeostasis-associated（Enho） gene encodes a secreted protein, adropin, which regulates the expression of hepatic lipogenic genes and adipose tissue peroxisome proliferator-activated receptor γ, a major regulator of lipogenesis. In the present study, the porcine（Sus scrofa） homologue of the Enho gene, which was named p Enho, was amplified by reverse transcriptase polymerase chain reaction（RT-PCR） using oligonucleotide primers derived from in silico sequences. The gene sequence was submitted into the Gen Bank of NCBI, and the access number is GQ414763. The p Enho encodes a protein of 76 amino acids which shows 75% similarity to Homo sapiens adropin. The expression profile of p Enho in tissues（liver, muscle, anterior jejunum, posterior jejunum, and ileum） was determined by quantitative real-time RT-PCR. p Enho was localized on porcine chromosome 10 and no introns were found. In conclusion, p Enho was cloned and analysed with the aim of increasing knowledge about glucose and lipid metabolism in piglets and helping to promote the health and growth of piglets through adropin regulation.

A novel large deletion mutation of FERMT1 gene in a Chinese patient with Kindler syndrome

Kindler syndrome （KS; OMIM 173650） is a rare autosomal recessive skin disorder, which results in symptoms including blistering, epidermal atrophy, increased risk of cancer, and poor wound healing. The majority of mutations of the disease-determining gene （FERMT1 gene） are single nucleotide substitutions, including missense mutations, nonsense mutations, etc. Large deletion mutations are seldom reported. To determine the mutation in the FERMT1 gene associated with a 7-year-old Chinese patient who presented clinical manifestation of KS, we performed direct sequencing of all the exons of FERMT1 gene. For the exons 2-6 without amplicons, we analyzed the copy numbers using quantitative real-time polymerase chain reaction （qRT-PCR） with specific primers. The deletion breakpoints were sublocalized and the range of deletion was confirmed by PCR and direct sequencing. In this study, we identified a new 17-kb deletion mutation spanning the introns 1-6 of FERMT1 gene in a Chinese patient with severe KS phenotypes. Her parents were carriers of the same mutation. Our study reported a newly identified large deletion mutation of FERMT1 gene involved in KS, which further enriched the mutation spectrum of the FERMT1 gene.

Transcription:the epicenter of gene expression

The complexity of a living organism is not driven by gene number but gene regulation. Controlling which genes to express and to what extent dictates the subsequent cell identity. Transcription, the critical initial stage in gene expression, is regulated delicately to maintain the cell status. Recent developments in the genomic approaches provided unparalleled coverage of the study of transcription. Still, basic molecular biology and biochemistry are providing mechanistic insights into how the regulation is achieved. In this feature “Regulation of transcription： mechanisms and biological functions”, the latest advances in epigenetics, mRNA processing, RNA quality control, and human immunodeficiency virus （HIV） transactivation are discussed.

Establishment of reference mandibular plane for anterior alveolar morphology evaluation using cone beam computed tomography

To propose a method of establishing the reference mandibular plane(MP),which could be reestablished according to the coordinates of the reference points,and then facilitate the assessment of anterior alveolar morphology using cone beam computed tomography(CBCT),sixty patients with bimaxillary protrusion were randomly selected and CBCT scans were taken.The CBCT scans were transferred to Materialism's interactive medical image control system 10.01(MIMICS 10.01),and three dimensional models of the entire jaws were constructed.Reference points determining the reference MP were positioned in the coronal,axial,sagittal windows,and the points were exactly located by recording their coordinates in the interfaces of software.The reference MP provided high intra-observer reliability(Pearson's r 0.992 to 0.999),and inter-observer reliability(intra-class correlation coefficients(ICCs) 0.996 to 0.999).

Methylenetetrahydrofolate reductase C677T polymorphism predicts response and time to progression to gemcitabine-based chemotherapy for advanced non-small cell lung cancer in a Chinese Han population

Objective： The aim of this study was to evaluate the association between the methylenetetrahydrofolate reductase （MTHFR） C677T excision repair cross-complementation group 1 （ERCC1） genetic polymorphisms and the clinical efficacy of gemcitabine-based chemotherapy in advanced non-small cell lung cancer （NSCLC）. Methods： A total of 135 chemonaive patients with unresectable advanced NSCLC were treated with gemcitabine/platinum regi- mens. The polymorphisms of MTHFR C677T, ERCC1 C8092A, and ERCC1 Cl18T were genotyped using the TaqMan methods. Results： The overall response rate was 28.9%. Patients with MTHFR CC genotype had a higher rate of objective response than patients with variant genotype （TT or CT） （41.2% versus 19.1%, P=0.01 ）. Median time to progression （TTP） of patients with MTHFR CC genotype was longer than that of patients with variant genotype （7.6 months versus 5.0 months, P=0.003）. No significant associations were obtained between ERCC1 C118T and C8092A polymorphisms and both response and survival. Conclusions： Our data suggest the value of MTHFR C677T polymorphism as a possible predictive marker of response and TTP in advanced NSCLC patients treated with gemcitabine/platinum.