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Abstract Background Gallbladder cancer is a major health concern in China, and awareness of the associated incidence and mortality rates is particularly important given the aging population. Objective To determine trends in gallbladder cancer incidence and mortality rates over 12 years and quantitatively analyze the influence of demographic factors on these rates in China. Methods We performed a retrospective study of 98,860 Chinese citizens using the Chinese Cancer Registry, a national database. Gallbladder cancer incidence and mortality data pertaining to patients treated between 2005 and 2017 were collected. Joinpoint regression models were used to estimate the annual percentage change (APC) and average APC (AAPC). We used age-period-cohort analyses and decomposition methods to investigate differing trends in incidence and mortality. Results The age-standardized gallbladder cancer incidence and mortality rates in China trended downward between 2005 and 2017, with AAPCs of -2.023% and -1.603%, respectively. Coefficients of age effect for incidence rate increased with age up to 70 years and peaked at 70–79 years, while coefficients of age effect for the mortality rate showed a consistent increase with age. Both coefficients of period for incidence and mortality rates increased in more recent periods; in terms of the cohort effect, coefficients of cohort for rates generally decreased in later birth years but showed a partial rise between 1982 and 1996. The crude incidence rates of gallbladder cancer according to demographic and non-demographic factors were 626.09% and -526.09% respectively (366.23% and -266.23% among men, and 6068.93% and -5968.93% among women, respectively). The rates were 543.01% and -443.01%, respectively, in urban areas and were 68.22% and 31.78%, respectively, in rural areas. The mortality rates according to demographic and non-demographic factors were -495.93% and 595.93%, respectively (-1763.10% and -1863.10% for men and -270.56% and -370.56% for women, respectively). These rates were -930.33% and 1030.33%, respectively, in urban areas and were 101.48% and -1.48%, respectively, in rural areas. Conclusions The overall standardized gallbladder cancer incidence and mortality rates in China are trending downward, but not sufficiently so. Proper living and eating habits should be encouraged while exploring the establishment of long-term, standardized gallbladder cancer screening programs.

Cell transfection is a specialized technique for introducing exogenous molecules, such as RNA and DNA, into cells and has become a prevalent method in gene functionality studies. The most frequently employed cell transfection methods include electroporation, viral infection, and liposome transfection. Liposome transfection is a prevalent technique in molecular biology for studying gene function and regulation. Liposome transfection has the advantages of wide applicability, superior efficiency, and simple operation. However, most liposome transfection reagents on the market are designed based on mammalian cells. Cell membrane composition, temperature adaptation range, and oxygen demand in teleost cells differ substantially from those in mammalian cells. Accordingly, the efficiency of liposome transfection in teleost cells is limited, and obtaining stable transfected cells directly through screening is challenging. Therefore, after the transfection of conventional transient expression vectors into teleost cells via liposomes, most of the plasmid copies tend to vanish due to cell proliferation after the duration of screening. Furthermore, the insignificant number of cells retaining plasmids makes it difficult to correctly integrate plasmids into the genome sequence, which hinders screening for stable cell lines. In this study, we investigated a method of rapidly obtaining stable transfected cells of Japanese flounder by transfecting the Tol2 transposon recombinant vector with liposomes. The CMV-EGFP-pminiTol2 vector was modified from the pminiTol2 plasmid, sustaining the functionality of the Tol2 transposable element to facilitate the highly efficient insertion of target genes into genomic DNA. The CMV-EGFP-pminiTol2 plasmid was highly suitable for stable transformation screening in Japanese flounder cells. This advantageous trait was mainly attributed to the following four aspects: (1) The CMV-EGFP-pminiTol2 plasmid carries autonomous transposon elements that can significantly enhance the integration ability of target genes. (2) The CMV-EGFP-pminiTol2 plasmid carries EGFP, allowing researchers to visualize transfection and screening effects directly via fluorescence microscopy. (3) The CMV-EGFP-pminiTol2 plasmid incorporates a neomycin resistance selection marker, which can be used to quickly and successfully screen transfected cells after G418 screening. (4) The multiple cloning site (MCS) region of the CMV-EGFP-pminiTol2 plasmid is mutated to retain two unique cleavage sites (XhoI and XmaI); plasmids can be linearized through these two sites, allowing the target gene to be inserted using homologous recombinase cloning. CMV-EGFP-pminiTol2 and pCS-TP plasmids were co-transfected into flounder cells via liposome encapsulation, and stable transfected cells were obtained through neomycin (800 μg/mL) screening for 2–3 weeks. After 2 weeks of screening, cells were stably cultured for 2–3 weeks without G418 screening. Finally, the EGFP fluorescence of the cells was observed using an inverted fluorescence microscope to confirm the transfection efficiency of the cells. The results revealed that the CMV-EGFP-pminiTol2 plasmid-transfected experimental group successfully expressed EGFP in all cells after G418 selection. In the pEGFP-C1- or pEGFP-N1-transfected control group, only a minor fraction of cells (≤10%) sustained EGFP expression after G418 screening. The CMV-EGFP- pminiTol2 vector is therefore suitable for the preparation of stable transgenic cells in Japanese flounder, offering numerous advantages, including reduced screening time, stable exogenous gene expression, simplified operations, and relatively low cost. Consequently, researchers can insert their desired target genes into the CMV-EGFP-pminiTol2 vector to construct stable transformed cells for investigations such as functional analysis and assessments of subcellular localization and long-term regulation. This study describes a novel approach to rapidly screening stably transformed flounder cells using the Tol2 transposon system. The results can be applied to molecular biology research in flounder and provide novel insights for constructing and screening stable transformations in teleost cells.

The razor clam (Sinonovacula constricta, Class Bivalvia) is a kind of burial filter-feeding shellfish. Salinity fluctuation is an important source of pressure for water habitats. High salinity in some coastal areas of Shandong and Jiangsu impact the survival and germplasm conservation of razor clam. To study the ecological behavior response of S. constricta to high salt culture environment, two populations of razor clams were used, including "Shenzhe No.1" population (SZSC) and a natural population (ZRSC). The semi-lethal salinity level of each population was determined. The effects of control group (20) and high salinity (24, 28, 32) on burrowing and feeding behavior of razor clams were studied. The differences in burrowing indices and feeding physiology between the two populations were compared. In the burrowing behavior experiment, two groups were set; razor clams from the temporary pond were put into each salinity group to start the experiment, while the other group of razor clams were stressed under each salinity condition for 24 h and then put into each salinity group to start the experiment. The results showed that the 120 h LC50 of SZSC was 34.04, while the 120 h LC50 of ZRSC was 32.04. The burrowing behavior of razor clams could be divided into four periods: The preparation period of shell closure, the period of axe foot movement, mud digging period, and the end period of mud diving. In the non-stressed group, the burrowing time of 50% (BT50) of SZSC was significantly higher than that of ZRSC (P0.05). The BT50 of SZSC was closer to that of the control group at 24 and 28 salinity, and the vitality of SZSC was significantly better than that of ZRSC after 24 h of salinity stress. Under high salinity, the distribution of SZSC in mud was more concentrated than that in ZRSC, and the burrowing depth was shallower. In terms of feeding physiology, the feeding rate of SZSC was significantly higher than that of ZRSC under high salinity (P

The potential of Euglena gracilis for carbon sequestration offers significant opportunities in the capture and utilization of carbon dioxide (CO2). In this study, a mutant LE-ZW of E. gracilis, capable of efficient growth and carbon sequestration, was obtained through ultraviolet mutagenesis combined with high carbon acclimation. Subsequently, the potential of LE-ZW for carbon assimilation was systematically analyzed. The results demonstrated that the cell density of the LE-ZW was 1.33 times that of the wild type and its carbon sequestration efficiency was 6.67 times that of the wild type when cultured at an optimal CO2 concentration of 5% until day 10. At this time, most key enzyme genes associated with the photosystem membrane protein complex, photosynthetic electron transport chain, antenna protein, and carbon fixation were up-regulated in mutant LE-ZW. Furthermore, after 10 days of culture under 10% CO2, the cell density and carbon sequestration efficiency of LE-ZW reached 1.10 times and 1.54 times of that under 5% CO2, respectively. Transcriptome analysis revealed significant up-regulation of key enzyme genes associated with carbon fixation, central carbon metabolism, and photosynthesis in LE-ZW under a 10% CO2 concentration. Physiological indices such as the amount of oxygen evolution, the values of Fv/Fm, the expression levels of photosynthetic protein genes and the enzyme activity of key enzymes related to photosynthetic carbon assimilation were corroborated by transcriptome data, elucidating that the mutant LE-ZW exhibited augmented photosynthetic carbon sequestration capacity and metabolic activity, thereby demonstrating robust adaptability to a high-carbon environment. This research contributes to a deeper understanding of the carbon assimilation mechanism in photosynthetic protists under elevated CO2 concentrations.

Androstenedione (ADSD) is one of the widely detected androgens in diverse aquatic environments. However, there were few reports on the molecular mechanism of Chlorella vulgaris exposure to ADSD. In our previous research, we have investigated the genes associated with chlorophyll metabolism in Chlorella vulgaris response to ADSD. In this study, we focus on continuously up-regulated genes to explore the mechanism underlying Chlorella vulgaris resistance to ADSD toxicity. Chlorella vulgaris was exposed to ADSD with five concentration gradients. The continuously up-regulated genes were enriched by Series Test of Cluster (STC) analysis and verified by qRT-PCR. Microalgae Super Oxidase Dimutase (SOD) and Microalgae Malonic dialdehyde (MDA), two indicators of oxidative stress, were determined by ELISA after exposure to ADSD. The results showed that ADSD can stimulate the production of extracellular polymeric substances (EPS) and lead to enlargement in the cell body of Chlorella vulgaris. In addition, steroid biosynthesis and oxidoreductase activity processes were consistently up-regulated upon exposure to ADSD. In conclusion, our study highlighted the crucial role of phenotypic modification, hormone synthesis, and redox mechanisms in protecting Chlorella vulgaris cells from the harmful effects of ADSD contamination.

Chemical composition is a critical factor for determining the efficacy of any traditional Chinese medicine (TCM) and can be used as an indicator of commercial quality. To develop a new strategy for discovering potential quality markers (Q-markers) of TCM by integrating ultra-performance liquid chromatography-Q-extractive orbitrap/mass spectrometry (UPLC-Q-Extractive Orbitrap/MS), chemometric analysis, and network pharmacology, using Schisandra chinensis (Turcz.) Baill. (S. chinensis) as an example. The chemical profiling of S. chinensis was performed using UPLC-Q-Extractive Orbitrap/MS, followed by identification of hepatoprotective Q-markers through a comprehensive understanding of chemometric analysis and virtual target prediction of network pharmacology. Six compounds were considered potent candidates for Q-markers, which were identified as schisandrol A (6), angeloylgomisin H (10), schisantherin A (17), schisantherin B (18), schisandrin A (23), and schisandrin C (26). All Q-markers exhibited significant hepatoprotective activity, as evidenced by in vitro experiments. Subsequently, a method for simultaneous quantification was established and employed to analyse seven batches of S. chinensis. Therefore, the integrated approach of UPLC-Q-Extractive Orbitrap/MS, chemometrics, and network pharmacology proved to be an effective strategy for the discovery of Q-markers that can assist in assessing the overall chemical consistency of samples and provide a basis for quality evaluation of the material basis of S. chinensis.

Alzheimer’s disease (AD) is a critical health issue worldwide that has a negative impact on patients’ quality of life, as well as on caregivers, society, and the environment. Positron emission tomography (PET)/computed tomography (CT) and neuropsychological scales can be used to identify AD and mild cognitive impairment (MCI) early, provide a differential diagnosis, and offer early therapies to impede the course of the illness. However, there are few reports of large-scale 11C-PIB-PET/CT investigations that focus on the pathology of AD and MCI. Therefore, further research is needed to determine how neuropsychological test scales and PET/CT measurements of disease progression interact.

Digestates from different anaerobic digesters are promising substrates for microalgal culture, leading to effective wastewater treatment and the production of microalgal biomass. However, further detailed research is needed before they can be used on a large scale. The aims of this study were to investigate the culture of Chlorella sp. in DigestateM from anaerobic fermentation of brewer’s grains and brewery wastewater (BWW) and to explore the potential use of the biomass produced under different experimental conditions, including diverse cultivation modes and dilution ratios. Cultivation in DigestateM initiated from 10% (v/v) loading, with 20% BWW, obtained maximum biomass production, reaching 1.36 g L−1 that was 0.27g L−1 higher than 1.09 g L−1 of BG11. In terms of DigestateM remediation, the maximum removal of ammonia nitrogen (NH4+-N), chemical oxygen demand, total nitrogen, and total phosphorus reached 98.20%, 89.98%, 86.98%, and 71.86%, respectively. The maximum lipid, carbohydrate, and protein contents were 41.60%, 32.44%, and 27.72%, respectively. The growth of Chlorella sp. may be inhibited when the Y(II)–Fv/Fm ratio is less than 0.4.

Catechins are the major flavor substances in teas, which have a variety of health effects; however, high catechin and high sensory quality are a pair of contradictions that are difficult to coordinate. To explore the processing procedure with high catechins and high sensory quality, a single-factor processing experiment was carried out over the processing production of oolong tea. Combined with orthogonal partial least square discriminant analysis (OPLS-DA), correlation analysis, and principal component analysis (PCA), the optimal production procedure for oolong tea is as follows: red light withering for 8 h, leaf rotating for 10 min with a total standing time for 8 h, drum roasting for 5 min at 290 °C, low-temperature rolling (flattening at 4 °C for 5 min, without pressure for 1 min and under pressure for 5 min), microwave drying (800 W for 7.5 min). This study demonstrates a significant increase in the retention of catechins, which contributes to the mellow and brisk tastes of oolong tea, addressing the challenge of catechin content and sensory quality. Our study provides a novel insight into the relationship between the oolong tea processing and flavor formation.

As the main flavor components of tea, the contents of epigallocatechin-3-gallate (EGCG), theanine and caffeine are regulated by ambient temperature. However, whether the biosynthesis of EGCG, theanine and caffeine in response to temperature is regulated by endogenous hormones and its mechanism is still unclear. In this study, tea cuttings cultivated in the phytotron which treated at different temperatures 15℃, 20℃, 25℃ and 30℃, respectively. The UPLC and ESI-HPLC-MS/MS were used to determine the contents of EGCG, theanine, caffeine and the contents of phytohormones in one leaf and a bud. The results showed that indoleacetic acid (IAA), gibberellin 1(GA1) and gibberellin 3 (GA3) were significantly correlated with the content of EGCG; Jasmonic acid (JA), jasmonate-isoleucine (JA-Ile) and methyl jasmonate (MeJA) were strongly correlated with theanine content; IAA, GA1 and gibberellin 4 (GA4) were significantly correlated with caffeine content at different temperatures. In order to explore the internal intricate relationships between the biosynthesis of these three main taste components, endogenous hormones, and structural genes in tea plants, we used multi-omics and multidimensional correlation analysis to speculate the regulatory mechanisms: IAA, GA1 and GA3 up-regulated the expressions of chalcone synthase (CsCHS) and trans-cinnamate 4-monooxygenase (CsC4H) mediated by the signal transduction factors auxin-responsive protein IAA (CsIAA) and DELLA protein (CsDELLA), respectively, which promoted the biosynthesis of EGCG; IAA, GA3 and GA1 up-regulated the expression of CsCHS and anthocyanidin synthase (CsANS) mediated by CsIAA and CsDELLA, respectively, via the transcription factor WRKY DNA-binding protein (CsWRKY), and promoted the biosynthesis of EGCG; JA, JA-Ile and MeJA jointly up-regulated the expression of carbonic anhydrase (CsCA) and down-regulated the expression of glutamate decarboxylase (CsgadB) mediated by the signal transduction factors jasmonate ZIM domain-containing protein (CsJAZ), and promoted the biosynthesis of theanine; JA, JA-Ile and MeJA also jointly inhibited the expression of CsgadB mediated by CsJAZ via the transcription factor CsWRKY and AP2 family protein (CsAP2), which promoted the biosynthesis of theanine; IAA inhibited the expression of adenylosuccinate synthase (CspurA) mediated by CsIAA via the transcription factor CsWRKY; GA1 and gibberellin 4 (GA4) inhibited the expression of CspurA mediated by CsDELLA through the transcription factor CsWRKY, which promoted the biosynthesis of caffeine. In conclusion, we revealed the underlying mechanism of the biosynthesis of the main taste components in tea plant in response to temperature was mediated by hormone signal transduction factors, which provided novel insights into improving the quality of tea.

Bisphenol A (BPA) can induce complex regulatory mechanisms in many aquatic organisms, and it is difficult to find a suitable analytical method to efficiently enrich key genes responding to BPA exposure. In this study, zebrafish embryo transcriptomic data were obtained from two types of different BPA exposure methods. After BPA exposure, three differential gene enrichment methods were used jointly to identify up-regulated genes or pathways in zebrafish embryo larvae. The results showed that the systemic lupus erythematosus signaling pathway was significantly enriched in all BPA exposure groups. It was also noteworthy that most of the up-regulated genes in systemic lupus erythematosus signaling were histones. In conclusion, this study suggested that autoimmunity signaling was the most common important pathway in zebrafish embryo-larvae response to different BPA exposures, and histones may play a key role in response to low-concentration BPA.

Sustainable light energy from ambient environment has attracted particular attention to meet the ever-growing need of small-scale electronics. The modulation of intercorrelated thermal and electronic transport is one of the crucial aspects for reliable photothermoelectric electronics. Herein, a defect-promoted photothermoelectric effect is demonstrated in densely aligned ZnO nanorod array with rich lattice defects. The defect-rich ZnO device delivers high electrical conductivity and large Seebeck coefficient to enable significant improvement of photothermoelectric energy conversion and self-powered photodetection. The position sensitivity reaches approximately 0.19 mV mm−1, and the temperature gradient induced electric field makes up for the suppression in the photothermoelectric process. The synergism between intrinsic defects and extra temperature field plays an important role in promoting the photothermoelectric properties of dense ZnO nanorod array. This study is interesting for interpreting the thermo-phototronic phenomena as well as demonstrating the possibility of defect engineering and phonon engineering to enable highly efficient light energy scavenging and self-powered photodetection.

Abstract Background Abnormalities of lymphocyte subsets and cytokine profiles have been observed in most patients with coronavirus disease (COVID-19). Here, we explore the role of lymphocyte subsets and cytokines on hospital admission in predicting the severity of COVID-19. Methods This study included 214 patients with COVID-19 who were treated at Chongqing University Three Gorges Hospital from January 19, 2020 to April 30, 2020. Any mutants were not detected in the studied patients. Patients were divided into non-intensive care unit (ICU) (mild/moderate) group and ICU (severe/critical) group, according to the severity of the disease. Clinical and laboratory data, including peripheral lymphocyte subsets and cytokines, were analyzed and compared. Logistic regression was used to analyze the predictive factors for ICU admission. Receiver operating characteristic (ROC) curves were drawn to evaluate the predictive value of selected indicators for the severity of COVID-19. Results Of the 214 patients enrolled, 161 were non-ICU patients and 53 were ICU patients. Lymphopenia was observed in nearly all of ICU patients (96.2%) and 84.5% of non-ICU patients on hospital admission. The absolute number of lymphocytes, CD3+ T cells, CD4+ T cells, CD8+ T cells, CD19+ B cells, and natural killer (NK) cells were lower in ICU group (659.00 × 106/L, 417.00 × 106/L, 261.00 × 106/L, 140.00 × 106/L, 109.00 × 106/L, 102.00 × 106/L, respectively) than in non-ICU group (1063.00 × 109/L, 717.00 × 106/L, 432.00 × 106/L, 271.00 × 106/L, 133.00 × 106/L, 143.00 × 106/L, respectively). Interleukin (IL)-6 was significantly higher in ICU patients than in non-ICU patients (18.08 pg/mL vs. 3.13 pg/mL, P

The slotted disk spring has been widely used given its nonlinear characteristics. However, the number of studies on them is limited, and the influences of nonuniform thickness have not been investigated theoretically and experimentally. Additionally, the hysteresis caused by friction has been neglected in most studies. By introducing the symmetric friction condition and the dimensionless thickness variation parameter, the present study establishes a mathematical model for slotted disk springs with linearly graded thickness. By treating the slotted segment as a number of cantilever beams with both a linearly gradient thickness and width, a new analytical formula is developed to characterize the load–deflection of slotted disk springs based on the moment equilibrium equation. The proposed model allows the direct quantification of the influences of the thickness variation parameter and the symmetric friction condition on the load–deflection characteristics. To validate the proposed model, slotted disk springs with different parameter configurations are designed and tested. Comparisons between measured and theoretical results illustrate that the proposed model can effectively describe the load–deflection characteristics of slotted disk springs with both uniform thickness and linearly gradient thickness. Due to the hypothetic linear variable width of slotted segments, the proposed model has a better performance in predicting slotted disk springs with rectangular slots.

The quality of Schisandra chinensis (Turcz.) Baill. (S. chinensis) is principally attributed to lignan compounds. In this paper, a simple and rapid strategy for simultaneous extraction and determination of 10 lignans from S. chinensis was established through matrix solid-phase dispersion (MSPD) assisted by diol-functionalized silica (Diol). The experimental parameters for MSPD extraction were screened using the response surface methodology (RSM). Diol (800 mg) was used as a dispersant and methanol (MeOH, 85%, v/v) as an eluting solvent (10 mL), resulting in a high extraction efficiency. MSPD extraction facilitated the combination of extraction and purification in a single step, which was less time-consuming than and avoided the thermal treatment involved in traditional methods. The simultaneous qualification and quantification of 10 lignans was achieved by combining MSPD and high-performance liquid chromatography (HPLC). The proposed method offered good linearity and a low limit of detection starting from 0.04 (schisandrin C) to 0.43 μg/mL (schisantherin B) for lignans, and the relative standard deviation (RSD, %) values of precision were acceptable, with a maximum value of 1.15% (schisantherin B and schisanhenol). The methodology was successfully utilized to analyze 13 batches of S. chinensis from different cultivated areas of China, which proved its accuracy and practicability in the quantitative analysis of the quality control of S. chinensis.

Abstract Background Chemoresistance influences the therapeutic effect of cisplatin and remains a major obstacle to its clinical use. MicroRNAs are associated with drug resistance of various tumors. However, the association between microRNAs and cisplatin in lung cancer remains largely unclear. Methods MicroRNA expression profile was identified by microRNA microarray between the lung cancer cisplatin‐sensitive cell line A549 (A549) and cisplatin‐resistant cell line A549/DDP (A549/DDP) and confirmed by quantitative real‐time‐PCR (qRT‐PCR). In vitro loss‐ and gain‐of‐function studies were performed to reveal the biological function of miR‐192 and related mechanism of the microRNA‐192/NKRF axis in lung cancer cell cisplatin resistance. Results Increased miR‐192 expression was detected in A549/DDP cells compared to A549. High miR‐192 expression significantly suppressed apoptosis, enhanced proliferation, and conferred resistance to cisplatin in lung cancer cells. NF‐κB repressing factor (NKRF), which is involved in the regulation of the NF‐κB signaling pathway, was identified as a direct target of miR‐192. Overexpression of miR‐192 significantly increased the nuclear protein amount and transcriptional activation of NF‐κB and expression of cIAP1, cIAP2, Bcl‐xl and XIAP, whereas decreased miR‐192 expression did the opposite. Inhibition of the NF‐κB signal pathway by curcumin reversed the effect of upregulation of miR‐192 on proliferation, apoptosis and cisplatin‐resistance in lung cancer cells. These results indicated that miR‐192/ NKRF axis enhances the cisplatin resistance of lung cancer cells through activating the NF‐κB pathway in vitro. Conclusions MiR‐192 plays a crucial role in cisplatin‐resistance of lung cancer cells. Thus, MiR‐192 may represent a therapeutic target for overcoming resistance to cisplatin‐based chemotherapy in lung cancer.

Abstract Background MicroRNAs (miRNAs) have been reported to play significant roles in non-small-cell lung cancer (NSCLC). However, the roles of microRNA (miR)-1915-3p in NSCLC remain unclear. In this study, we aimed to explore the biological functions of miR-1915-3p in NSCLC. Methods The expression of miR-1915-3p and SET nuclear proto-oncogene (SET) in NSCLC tissues were examined by quantitative real-time PCR (qRT-PCR). Migratory and invasive abilities of lung cancer were tested by wound healing and transwell invasion assay. The direct target genes of miR-1915-3p were measured by dual-luciferase reporter assay and western blot. Finally, the regulation between METTL3/YTHDF2/KLF4 axis and miR-1915-3p were evaluated by qRT-PCR, promoter reporter assay and chromatin immunoprecipitation (CHIP). Results miR-1915-3p was downregulated in NSCLC tissues and cell lines, and inversely associated with clinical TNM stage and overall survival. Functional assays showed that miR-1915-3p significantly suppressed migration, invasion and epithelial-mesenchymal transition (EMT) in NSCLC cells. Furthermore, miR-1915-3p directly bound to the 3′untranslated region (3′UTR) of SET and modulated the expression of SET. SET inhibition could recapitulate the inhibitory effects on cell migration, invasion and EMT of miR-1915-3p, and restoration of SET expression could abrogate these effects induced by miR-1915-3p through JNK/Jun and NF-κB signaling pathways. What’s more, miR-1915-3p expression was regulated by METTL3/YTHDF2 m6A axis through transcription factor KLF4. Conclusions These findings demonstrate that miR-1915-3p function as a tumor suppressor by targeting SET and may have an anti-metastatic therapeutic potential for lung cancer treatment.

The habitat occupied by flatfish fry differs considerably in light regime from that of the adult. The diversity of opsins and their expression patterns appear greatest for flatfishes to experience variable light environments. Yet, opsin repertoires and expression patterns in this group of fishes are poorly described. To understand how the visual system has adapted to such changes, we unveil that Japanese flounder (Paralichthys olivaceus) has a visual system adapted to a benthic environment by fine-tuning paralogous opsins (SWS2A and RH2) for wavelength shift and regulated expression. P. olivaceus express five basic opsin genes (M/LWS, SWS1, SWS2, RH1 and RH2) and gene-specific duplications were observed in RH2 and SWS2 paralogues. The expression of the three short-wavelength sensitive genes, SWS2Aα, SWS2Aβ, and SWS2B, is significantly elevated at the benthic stages, especially in SWS2Aβ a striking expression change is observed. The four middle-wavelength sensitive genes exhibit divergent expressions, the expression of RH2A-1 and RH2A-2 increased, while that of RH2B-1 and RH2B-2 decreased significantly from pelagic to benthic stage, especially RH2A-2. At present, changes at a total of 26 sites are known to have modified the λmax of various visual pigments during vertebrate evolution. Thus, these tuning site variations in our P. olivaceus are suspected to cause a green-shift in the λmax of SWS2Aα pigments and blue-shift in that of in RH2A-2. Together, our results suggest that RH2 and SWS2 opsin repertoires serve to optimize visual function under variable light environments by gene family duplications, differential expressions, and maximum absorption wavelength (λmax) variations.

During morphogenesis, epithelial monolayers actively alter their shape to create future body parts of the animal; this makes the epithelium one of the most active and critical structures in early animal development. While epithelia are often modeled as two-dimensional systems, real epithelia are not necessarily thin relative to cell cross section, and advances in 3D imaging have shown the possibility of substantial cell deformations in the third dimension, as well as differences in dynamics of the apical and basal surfaces indicative of three-dimensional coupling. With the importance of the third dimension in mind, we have developed a self-sculpting, three-dimensional model of epithelia whose dynamics are driven by active forces on its surface. We present a first, fundamental study for a reduced version of epithelia that investigates how surface forces affect its internal dynamics. Our model captures the 3D slab-like geometry of epithelia, viscoelasticity of tissue response, fluid surroundings, and driving from active surface forces. We represent epithelial tissue as a thick slab, a 3D continuum comprised of a Stokes fluid with an extra viscoelastic stress. Employing this model, we present both analytical and numerical solutions of the system and make quantitative predictions about cell shapes, cell dynamics, and the tissue's response to surface force in a three-dimensional setting. In particular, we investigate the implications of our model on the initiation of ventral furrow invagination and T1 transitions in Drosophila embryogenesis. In the former, we demonstrate the importance of fluid and geometric surroundings to drive invagination. In the latter, we show the limitations of surface forces alone to drive T1 transitions.

The razor clam, Sinonovacula constricta, is a commercially important bivalve. To date, there have been very limited quantitative genetic studies of this species, especially in different aquaculture environments. This study is the first report of genotype by environment (G × E) interactions for growth traits of razor clams from outdoor pond (OP) and semi-natural environments (SNE). Over a 2-month period, the data for shell length (SL), shell height (SH), shell width (SW), and body weight (BW) of razor clams were recorded for 1 month, and marked as Data 1 and Data 2. Phenotypic data of 1627 individual clams from 17 full-sib families were used for the analysis. The overall heritabilities (h2) for all growth traits were from low to moderate, ranging from 0.127 to 0.230. The h2 of SNE was higher than that of OP (0.243–0.423 vs. 0.101–0.285). Heritabilities for growth traits indicated that these traits could be effectively selected in both aquaculture environments. Overall, both phenotypic (rp) and genetic (rg) characteristics showed high positive correlations among the growth traits. However, the rg and rp were highly variable in OP, ranging from 0.435 to 0.973 and from 0.597 to 0.874, respectively. The rg of SL, SH, SW, and BW between the two aquaculture environments for Data 1 were 0.867, 0.682, 0.651, and 0.844, respectively. The rg ranged from 0.274 to 0.619 for Date 2. These results indicated significant G × E interactions among families at different sites. This study therefore provided optimal guidelines to select breeding programs for razor clams reared in different aquaculture environments.

Gibberellic acids had been proven to improve the fruit quality and storability by delaying deterioration and maintaining the antioxidant system. In this study, the effect of GA3 spraying at different concentrations (10, 20, and 50 mg L−1) on the quality of on-tree preserved ‘Shixia’ longan was examined. Only 50 mg L−1 GA3 significantly delayed the decline of soluble solids (22.0% higher than the control) and resulted in higher total phenolics content (TPC), total flavonoid content (TFC), and phenylalanine ammonia-lyase activity in pulp at the later stages. The widely targeted metabolome analysis showed that the treatment reprogrammed secondary metabolites and up-regulated many tannins, phenolic acids, and lignans during the on-tree preservation. More importantly, the preharvest 50 mg L−1 GA3 spraying (at 85 and 95 days after flowering) led to significantly delayed pericarp browning and aril breakdown, as well as lower pericarp relative conductivity and mass loss at the later stages of room-temperature storage. The treatment also resulted in higher antioxidants in pulp (vitamin C, phenolics, and reduced glutathione) and pericarp (vitamin C, flavonoids, and phenolics). Therefore, preharvest 50 mg L−1 GA3 spraying is an effective method for maintaining the quality and up-regulating antioxidants of longan fruit during both on-tree preservation and room-temperature storage.

Abstract Background Ovarian tissue cryopreservation has a wide range of cancerous indications. Avoiding relapse becomes a specific concern that clinicians frequently encounter. The data about the comparative viability of cancer cells after cryopreservation are limited. This study aimed to evaluate the effect of cryopreservation on breast cancer cells. Methods We used in-vitro cultured ZR-75-1 and MDA-MB-231 cell lines. Cell samples of each lineage were distributed into the non-intervened and cryopreserved groups. The cryopreservation procedures comprised programmed slow freezing followed by thawing at 100 °C, 60 s. Biological phenotypes and the related protein markers were compared between the two groups. The EVOS FL Auto 2 Cell Image System was used to monitor cell morphology. Cell proliferation, motility, and penetration were characterized by CCK-8, wound-healing, and transmembrane assay, respectively. The expression of Ki-67, P53, GATA3, E-cadherin, Vimentin, and F-Actin was captured by immunofluorescent staining and western blotting as the proxy measurements of the related properties. The chorioallantoic membrane (CAM) xenotransplantation was conducted to explore angiogenesis induced by cancer cells. Results After 5 days in vitro culture, the cell concentration of cryopreserved and non-intervened groups was 15.7 × 104 vs. 14.4 × 104cells/ml, (ZR-75-1, p > 0.05), and 25.1 × 104 vs. 26.6 × 104 cells/ml (MDA-MB-231, p > 0.05). Some cryopreserved ZR-75-1 cells presented spindle shape with filopodia and lamellipodia and dissociated from the cell cluster after cryopreservation. Both cell lines demonstrated increased cell migrating capability and invasion after cryopreservation. The expression of Ki-67 and P53 did not differ between the cryopreserved and non-intervened groups. E-cadherin and GATA3 expression downregulated in the cryopreserved ZR-75-1 cells. Vimentin and F-actin exhibited an upregulated level in cryopreserved ZR-75-1 and MDA-MB-231 cells. The cryopreserved MDA-MB-231 cells induced significant angiogenesis around the grafts on CAM with the vascular density 0.313 ± 0.03 and 0.342 ± 0.04, compared with that of non-intervened cells of 0.238 ± 0.05 and 0.244 ± 0.03, p < 0.0001. Conclusions Cryopreservation promotes breast cancer cells in terms of epithelial-mesenchymal transition and angiogenesis induction, thus increasing metastasis risk.

Abstract The Internet of Things (IoT) networks have become the infrastructure to enable the detection and reaction of anomalies in various domains, where an efficient sensory data gathering mechanism is fundamental since IoT nodes are typically constrained in their energy and computational capacities. Besides, anomalies may occur occasionally in most applications, while the majority of time durations may reflect a healthy situation. In this setting, the range, rather than an accurate value of sensory data, should be more interesting to domain applications, and the range is represented in terms of the category of sensory data. To decrease the energy consumption of IoT networks, this paper proposes an energy-efficient sensory data gathering mechanism, where the category of sensory data is processed by adopting the compressed sensing algorithm. The sensory data are forecasted through a data prediction model in the cloud, and sensory data of an IoT node is necessary to be routed to the cloud for the synchronization purpose, only when the category provided by this IoT node is different from the category of the forecasted one in the cloud. Experiments are conducted and evaluation results demonstrate that our approach performs better than state-of-the-art techniques, in terms of the network traffic and energy consumption.

Although the effects of phytohormones (mainly salicylic acid) on the storability of longan fruit have been reported, the relationship between postharvest hormone variation and signal transduction and storability remains unexplored. The basis of physiology, biochemistry, hormone content and signalling for the storability difference at room-temperature between ‘Shixia’ and ‘Luosanmu’ longan fruit were examined. ‘Luosanmu’ longan exhibited faster pericarp browning, aril breakdown and rotting during storage. ‘Luosanmu’ pericarp exhibited higher malondialdehyde but faster decreased total phenolics, flavonoid, glutathione, vitamin C, catalase activity and gene expression. Higher H2O2 and malondialdehyde but lower glutathione, glutathione-reductase and peroxidase activities, while higher activities and gene expressions of polygalacturonase, β-galactosidase and cellulose, lower covalent-soluble pectin, cellulose and hemicellulose but higher water-soluble pectin were observed in ‘Luosanmu’ aril. Lower abscisic acid and methyl jasmonate but higher expressions of LOX2, JAZ and NPR1 in pericarp, while higher abscisic acid, methyl jasmonate and salicylic acid together with higher expressions of ABF, JAZ, NPR1 and PR-1 in ‘Luosanmu’ aril were observed. In conclusion, the imbalance between the accumulation and scavenging of active oxygen in ‘Luosanmu’ longan might induce faster lipid peroxidation and senescence-related hormone signalling and further the polymerization of phenolics in pericarp and polysaccharide degradation in aril.

Adaptation can optimize information processing by allowing the visual system to always adjust to the environment. However, only a few studies have investigated how the visual system makes adjustments to repeatedly occurring changes in the input, still less about the related neural mechanism. Our previous study found that contrast adaptation attenuated after multiple daily sessions of repeated adaptation, which was explained by the habituation of either the adapter’s effective strength or the adaptation mechanisms. To examine the former hypothesis, in the present study we used the frequency tagging technique to measure the adapter-elicited steady-state visual evoked potential (SSVEP) amplitudes. Participants repeatedly adapted to the same contrast adapter in a top-up manner for six continuous days, which was called training of adaptation. The behavioral adaptation effect and SSVEP response to the trained adapter and an untrained control adapter were measured before and after training. The psychophysical results showed that the effect of adaptation in the trained condition significantly reduced after training, replicating our previous finding. Contradicting the prediction of the hypothesis that repeated adaptation attenuated the effective strength of the adapter, the SSVEP amplitude was unchanged after training, which was further confirmed by an equivalence test. Taken together, the results challenge the account of habituation of adapter in repeated adaptation, while leaving the account of habituation of adaptation mechanism to be tested.

Sex reversal induced by temperature change is a common feature in fish. Usually, the sex ratio shift occurs when temperature deviates too much from normal during embryogenesis or sex differentiation stages. Despite decades of work, the mechanism of how temperature functions during early development and sex reversal remains mysterious. In this study, we used Chinese tongue sole as a model to identify features from gonad transcriptomic and epigenetic mechanisms involved in temperature induced masculinization. Some of genetic females reversed to pseudomales after high temperature treatment which caused the sex ratio imbalance. RNA-seq data showed that the expression profiles of females and males were significantly different, and set of genes showed sexually dimorphic expression. The general transcriptomic feature of pesudomales was similar with males, but the genes involved in spermatogenesis and energy metabolism were differentially expressed. In gonads, the methylation level of cyp19a1a promoter was higher in females than in males and pseudomales. Furthermore, high-temperature treatment increased the cyp19a1a promoter methylation levels of females. We observed a significant negative correlation between methylation levels and expression of cyp19ala. In vitro study showed that CpG within the cAMP response element (CRE) of the cyp19a1a promoter was hypermethylated, and DNA methylation decreased the basal and forskolin-induced activities of cyp19a1a promoter. These results suggested that epigenetic change, i.e., DNA methylation, which regulate the expression of cyp19a1a might be the mechanism for the temperature induced masculinization in tongue sole. It may be a common mechanism in teleost that can be induced sex reversal by temperature.

Organ renewal is governed by the dynamics of cell division, differentiation and loss. To study these dynamics in real time, we present a platform for extended live imaging of the adult Drosophila midgut, a premier genetic model for stem-cell-based organs. A window cut into a living animal allows the midgut to be imaged while intact and physiologically functioning. This approach prolongs imaging sessions to 12–16 hr and yields movies that document cell and tissue dynamics at vivid spatiotemporal resolution. By applying a pipeline for movie processing and analysis, we uncover new and intriguing cell behaviors: that mitotic stem cells dynamically re-orient, that daughter cells use slow kinetics of Notch activation to reach a fate-specifying threshold, and that enterocytes extrude via ratcheted constriction of a junctional ring. By enabling real-time study of midgut phenomena that were previously inaccessible, our platform opens a new realm for dynamic understanding of adult organ renewal.

This paper proposes a novel permanent magnet (PM) motor for high torque performance, in which hybrid PM material and asymmetric rotor design are applied. The hybrid PM material is adopted to reduce the consumption of rare-earth PM because ferrite PM is assisted to enhance the torque production. Meanwhile, the rotor structure is designed to be asymmetric by shifting the surface-insert PM (SPM), which is used to improve the torque performance, including average torque and torque ripple. Moreover, the reasons for improvement of the torque performance are explained by evaluation and analysis of the performances of the proposed motor. Compared with SPM motor and V-type motor, the merit of high utilization ratio of rare-earth PM is also confirmed, showing that the proposed motor can offer higher torque density and lower torque ripple simultaneously with less consumption of rare-earth PM.

Background and objective It has been proven that increase of the awareness level of lung cancer prevention and control could enhance participation of lung cancer screening of lung cancer high risk group. The aim of this study is to investigate the awareness level of lung cancer prevention and control and the effect of individual characteristics on lung cancer awareness, and to provide evidence for comprehensive lung cancer prevention in high risk areas of lung cancer. Methods Staffs of Tianjin Dagang Oil Field who participate low dose CT (LDCT) lung cancer screening by cluster sampling or according to voluntary principle were surveyed, data of lung cancer awareness were collected by questionnaire. Results A total of 1,633 valid questionnaires were collected. The average age of respondents was 60.08±6.58. Most participants were males (82.2%) while female only accounted for 17.8%. The proportions of awareness about lung cancer in China, risk factors, screening methods and the knowledge of health examination were 64.5%, 77.1%, 43.7%, 49.6% respectively. Result of multiple logistic regression analysis showed that education level, smoking (pack-year), age, prior tuberculosis were the influencing factors of lung cancer awareness with adjusted Ors for education and age level as of 0.567 (95%CI: 0.439-0.733) and 1.373 (95%CI: 1.084-1.739) respectively. 80.3% of the participants can accept health examination once a year, while the ability to pay the medical expenses was not high. The influencing factors of health examination willingness were gender, age, income, the knowledge of lung cancer. Conclusion Education level and smoking affect the awareness of lung cancer prevention and control, health education for lung cancer should be conducted especially in population with low education level. Comprehensive lung cancer control in high risk areas should combined lung cancer screening, tobacco control and health education.

Following the two rounds of whole-genome duplication (WGD) during deuterosome evolution, a third genome duplication occurred in the ray-fined fish lineage and is considered to be responsible for the teleost-specific lineage diversification and regulation mechanisms. As a receptor-regulated SMAD (R-SMAD), the function of SMAD3 was widely studied in mammals. However, limited information of its role or putative paralogs is available in ray-finned fishes. In this study, two SMAD3 paralogs were first identified in the transcriptome and genome of Japanese flounder (Paralichthys olivaceus). We also explored SMAD3 duplication in other selected species. Following identification, genomic structure, phylogenetic reconstruction, and synteny analyses performed by MrBayes and online bioinformatic tools confirmed that smad3a/3b most likely originated from the teleost-specific WGD. Additionally, selection pressure analysis and expression pattern of the two genes performed by PAML and quantitative real-time PCR (qRT-PCR) revealed evidence of subfunctionalization of the two SMAD3 paralogs in teleost. Our results indicate that two SMAD3 genes originate from teleost-specific WGD, remain transcriptionally active, and may have likely undergone subfunctionalization. This study provides novel insights to the evolution fates of smad3a/3b and draws attentions to future function analysis of SMAD3 gene family.

Background and objective BAG genes (Bcl-2-associated athanogene) belong to a recently discovered multifunctional anti-apoptosis gene family that regulate various physiological processes which include apoptosis, tumorigenesis, neural differentiation, stress response and cell cycle and so on. The expression status of BAG family genes are related to certain tumor incidence and prognosis. The aim of this study is to explore the association of the BAG family gene expression status with the susceptibility of lung adenocarcinoma. Methods The gene expression data of BAG family genes from 29 cases of lung adenocarcinoma tissues and matched pericancerous lung tissess were generated by microarray chips. Cox regression was used to analyze the association between the expression of BAG family genes and the susceptibility of lung adenocarcinoma and the results were verified by GEO database. Results The expression levels of BAG-1, BAG-2, BAG-5 in cancer tissues were significantly downregulated compared with matched pericancerous lung tissues and were protective factors of lung adenocarcinoma (P < 0.05, OR < 1); while the expression level of BAG-4 in cancer tissues were remankably upregulated compared with the matched pericancerous lung tissues and was risk factor of lung adenocarcinoma (P < 0.05, OR > 1). Conclusion BAG-1, BAG-2, BAG-5 might be the potential protective factors while BAG-4 is possible risk factor of lung adenocarcinoma.

Autonomous vehicles are expected to play a key role in the future of urban transportation systems, as they offer potential for additional safety, increased productivity, greater accessibility, better road efficiency, and positive impact on the environment. Research in autonomous systems has seen dramatic advances in recent years, due to the increases in available computing power and reduced cost in sensing and computing technologies, resulting in maturing technological readiness level of fully autonomous vehicles. The objective of this paper is to provide a general overview of the recent developments in the realm of autonomous vehicle software systems. Fundamental components of autonomous vehicle software are reviewed, and recent developments in each area are discussed.

The purpose of this study was to determine the role of long-chain fatty acyl-CoA synthetase 4 (ACSL4) in breast cancer. Public databases were utilized to analyze the relationship between ACSL4 mRNA expression and the presence of steroid hormone and human epidermal growth factor receptor 2 (HER2) in both breast cancer cell lines and tissue samples. In addition, cell lines were utilized to assess the consequences of either increased or decreased levels of ACSL4 expression. Proliferation, migration, anchorage-independent growth and apoptosis were used as biological end points. Effects on mRNA expression and signal transduction pathways were also monitored. A meta-analysis of public gene expression databases indicated that ACSL4 expression is positively correlated with a unique subtype of triple negative breast cancer (TNBC), characterized by the absence of androgen receptor (AR) and therefore referred to as quadruple negative breast cancer (QNBC). Results of experiments in breast cancer cell lines suggest that simultaneous expression of ACSL4 and a receptor is associated with hormone resistance. Forced expression of ACSL4 in ACSL4-negative, estrogen receptor α (ER)-positive MCF-7 cells resulted in increased growth, invasion and anchorage independent growth, as well as a loss of dependence on estrogen that was accompanied by a reduction in the levels of steroid hormone receptors. Sensitivity to tamoxifen, triacsin C and etoposide was also attenuated. Similarly, when HER2-positive, ACSL4-negative, SKBr3 breast cancer cells were induced to express ACSL4, the proliferation rate increased and the apoptotic effect of lapatinib was reduced. The growth stimulatory effect of ACSL4 expression was also observed in vivo in nude mice when MCF-7 control and ACSL4-expressing cells were utilized to induce tumors. Our data strongly suggest that ACSL4 can serve as both a biomarker for, and mediator of, an aggressive breast cancer phenotype.