Dual-role transcription factors stabilize intermediate expression levels.

Precise control of gene expression levels is essential for normal cell functions, yet how they are defined and tightly maintained, particularly at intermediate levels, remains elusive. Here, using a series of newly developed sequencing, imaging, and functional assays, we uncover a class of transcription factors with dual roles as activators and repressors, referred to as condensate-forming level-regulating dual-action transcription factors (TFs). They reduce high expression but increase low expression to achieve stable intermediate levels. Dual-action TFs directly exert activating and repressing functions via condensate-forming domains that compartmentalize core transcriptional unit selectively. Clinically relevant mutations in these domains, which are linked to a range of developmental disorders, impair condensate selectivity and dual-action TF activity. These results collectively address a fundamental question in expression regulation and demonstrate the potential of level-regulating dual-action TFs as powerful effectors for engineering controlled expression levels.

Induction of mouse totipotent stem cells by a defined chemical cocktail.

In mice, only the zygotes and blastomeres from 2-cell embryos are authentic totipotent stem cells (TotiSCs) capable of producing all the differentiated cells in both embryonic and extraembryonic tissues and forming an entire organism1. However, it remains unknown whether and how totipotent stem cells can be established in vitro in the absence of germline cells. Here we demonstrate the induction and long-term maintenance of TotiSCs from mouse pluripotent stem cells using a combination of three small molecules: the retinoic acid analogue TTNPB, 1-azakenpaullone and the kinase blocker WS6. The resulting chemically induced totipotent stem cells (ciTotiSCs), resembled mouse totipotent 2-cell embryo cells at the transcriptome, epigenome and metabolome levels. In addition, ciTotiSCs exhibited bidirectional developmental potentials and were able to produce both embryonic and extraembryonic cells in vitro and in teratoma. Furthermore, following injection into 8-cell embryos, ciTotiSCs contributed to both embryonic and extraembryonic lineages with high efficiency. Our chemical approach to totipotent stem cell induction and maintenance provides a defined in vitro system for manipulating and developing understanding of the totipotent state and the development of multicellular organisms from non-germline cells.

Hidden Charge Order and Multiple Electronic Instabilities in EuTe4.

The rare-earth telluride compound EuTe4 exhibits a charge density wave (CDW) and an unconventional thermal hysteresis transition. Herein, we report a comprehensive study of the CDW states in EuTe4 by using low-temperature scanning tunneling microscopy. Two types of charge orders are observed at 4 K, including a newly discovered spindle-shaped pattern and a typical stripe-like pattern. As an exotic charge order, the spindle-shaped CDW is off-axis and barely visible at 77 K, indicating that it is a hidden order developed at low temperature. Based on our first-principles calculations, we reveal the origins of the observed electronic instabilities. The spindle-shaped charge order stems from a subsequent transition based on the stripe-like CDW phase. Our work demonstrates that the competition and cooperation between multiple charge orders can generate exotic quantum phases.

Coherent control on the generation and annihilation of a pseudospin-induced optical vortex in a honeycomb photonic lattice.

We experimentally investigate the coherently controllable generation and annihilation of a pseudospin-induced optical vortex in an optically induced honeycomb photonic lattice in a Λ-type 85Rb atomic vapor cell. Three Gaussian coupling beams are coupled into the atomic gases to form a hexagonal interference pattern, which can induce a honeycomb photonic lattice under electromagnetically induced transparency. Then, two probe beams interfere with each other to form periodical fringes and cover one set of sublattice in the honeycomb lattice, corresponding to excite the K or K' valleys in momentum space. By properly adjusting the experimental parameters, the generation and annihilation of the induced optical vortex can be effectively controlled. The theoretical simulations based on the Dirac and Schrödinger equations are performed to explore the underlying mechanisms, which will support the observations. The demonstrated properties of such controllable optical vortex may lay the foundation for the design of vortex-based optical devices with multidimensional tunability.

Non-Hermitian Delocalization in a Two-Dimensional Photonic Quasicrystal.

Theoretical and experimental studies suggest that both Hermitian and non-Hermitian quasicrystals show localization due to the fractal spectrum and to the transition to diffusive bands via exceptional points, respectively. Here, we present an experimental study of a dodecagonal photonic quasicrystal based on electromagnetically induced transparency in a Rb vapor cell. First, we observe the suppression of the wave packet expansion in the Hermitian case. We then discover a new regime, where increasing the non-Hermiticity leads to delocalization, demonstrating that the behavior in non-Hermitian quasicrystals is richer than previously thought.

Tetraspanin 1 regulates papillary thyroid tumor growth and metastasis through c-Myc-mediated glycolysis.

Papillary thyroid cancer (PTC) is the most common form of thyroid cancer and is characterized by its tendency for lymphatic metastasis, leading to a poor prognosis. Tetraspanin 1 (TSPAN1) is a member of the tetra-transmembrane protein superfamily and has been implicated in tumorigenesis and cancer metastasis in various studies. However, the role of TSPAN1 in PTC tumor development remains unclear. In this study, we aimed to investigate the impact of TSPAN1 on PTC cell behavior. Our results demonstrate that knockdown of TSPAN1 inhibits PTC cell proliferation, migration, and invasion, while overexpression of TSPAN1 has the opposite effect. These findings suggest that TSPAN1 might play a role in the tumorigenesis and invasiveness of PTC. Mechanistically, we found that TSPAN1 activates the ERK pathway by increasing its phosphorylation, subsequently leading to upregulated expression of c-Myc. Additionally, we observed that TSPAN1-ERK-c-Myc axis activation promotes glycolytic activity in PTC cells, as evidenced by the upregulation of glycolytic genes such as LDHA. Taken together, our findings indicate that TSPAN1 acts as an oncogene in PTC by regulating glycolytic metabolism. This discovery highlights the potential of TSPAN1 as a promising therapeutic target for PTC treatment. Further research in this area could provide valuable insights into the development of targeted therapies for PTC patients.

Loss Difference Induced Localization in a Non-Hermitian Honeycomb Photonic Lattice.

Non-Hermitian systems with complex-valued energy spectra provide an extraordinary platform for manipulating unconventional dynamics of light. Here, we demonstrate the localization of light in an instantaneously reconfigurable non-Hermitian honeycomb photonic lattice that is established in a coherently prepared atomic system. One set of the sublattices is optically modulated to introduce the absorptive difference between neighboring lattice sites, where the Dirac points in reciprocal space are extended into dispersionless local flat bands, with two shared eigenstates: low-loss (high-loss) one with fields confined at sublattice B (A). When these local flat bands are broad enough due to larger loss difference, the incident beam with its tangential wave vector being at the K point in reciprocal space is effectively localized at sublattice B with weaker absorption, namely, the commonly seen power exchange between adjacent channels in photonic lattices is effectively prohibited. The current work unlocks a new capability from non-Hermitian two-dimensional photonic lattices and provides an alternative route for engineering tunable local flat bands in photonic structures.

Prevalence of and risk factors for chronic kidney disease in ten metropolitan areas of China: a cross-sectional study using three kidney damage markers.

INTRODUCTION: To estimate the up-to-date prevalence of chronic kidney disease among the health check-up population in economically developed areas of China using estimated glomerular filtration rate, urinary albumin creatinine ratio, and kidney ultrasound. METHODS: Healthcare data from 38,093 subjects in 10 megalopolises of China who had an annual health check-up in 2021 were used. The overall and stratified prevalence of chronic kidney disease by sex, age, region and comorbidity group was reported. The association between chronic kidney disease and covariates of demographics, and comorbidities were analyzed in the multivariable-adjusted logistic regression model. RESULTS: A total of 3837 CKD cases were detected meeting any of the three CKD diagnostic criteria, with a crude prevalence of 10.1% in the study population. Using one criterion of decreased glomerular filtration rate, albuminuria and kidney structural abnormalities alone detected 204 (5.3%), 3289 (85.7%) and 563 (14.7%) cases, respectively. The addition of kidney ultrasound detected 427 (11.1%) structural abnormality cases without decreased GFR and albuminuria. The most common abnormalities were renal masses, hydronephrosis due to obstruction and congenital anomalies of kidney and urinary tract. Female, older age, low city-tier, hypertension, diabetes, obesity, hypertriglyceridemia as well as early disease stages such as pre-hypertension, impaired fasting glucose and overweight were significantly associated with chronic kidney disease. CONCLUSION: Kidney ultrasound helps to amplify the detection of CKD patients, which is a supplement to kidney function and urine protein.

Evolution of Electronic Structure in Pristine and Rb-Reconstructed Surfaces of Kagome Metal RbV3Sb5.

We report on in situ low-temperature (4 K) scanning tunneling microscope measurements of atomic and electronic structures of the cleaved surfaces of an alkali-based kagome metal RbV3Sb5 single crystals. We find that the dominant pristine surface exhibits Rb-1×1 structure, in which a unique unidirectional √3a0 charge order is discovered. As the sample temperature slightly rises, Rb-√3×1 and Rb-√3×√3 reconstructions form due to desorption of surface Rb atoms. Our conductance mapping results demonstrate that Rb desorption not only gives rise to hole doping but also reconstructs the electronic band structures. Surprisingly, we find a ubiquitous gap opening near the Fermi level in tunneling spectra on all the surfaces despite their large differences of hole-carrier concentration, indicating an orbital-selective band reconstruction in RbV3Sb5. The Rb desorption induced electronic reconstructions are further confirmed by our density functional theory calculations.

Method Validation for Multi-Pesticide Residue Determination in Chrysanthemum.

The chrysanthemum can be consumed in various forms, representing the "integration of medicine and food". Quantitative analysis of multi-pesticide residues in chrysanthemum matrices is therefore crucial for both product-safety assurance and consumer-risk evaluation. In the present study, a simple and effective method was developed for simultaneously detecting 15 pesticides frequently used in chrysanthemum cultivation in three matrices, including fresh flowers, dry chrysanthemum tea, and infusions. The calibration curves for the pesticides were linear in the 0.01-1 mg kg-1 range, with correlation coefficients greater than 0.99. The limits of quantification (LOQs) for fresh flowers, dry chrysanthemum tea, and infusions were 0.01-0.05 mg kg-1, 0.05 mg kg-1, and 0.001-0.005 mg L-1, respectively. In all selected matrices, satisfactory accuracy and precision were achieved, with recoveries ranging from 75.7 to 118.2% and relative standard deviations (RSDs) less than 20%. The validated method was then used to routinely monitor pesticide residues in 50 commercial chrysanthemum-tea samples. As a result, 56% of samples were detected with 5-13 pesticides. This research presents a method for the efficient analysis of multi-pesticide residues in chrysanthemum matrices.

Eye-Preserving Therapies for Advanced Retinoblastoma: A Multicenter Cohort of 1678 Patients in China.

PURPOSE: This study attempted to estimate the impact of eye-preserving therapies for the long-term prognosis of patients with advanced retinoblastoma with regard to overall survival and ocular salvage. DESIGN: Retrospective cohort study covering all 31 provinces (38 retinoblastoma treating centers) of mainland China. PARTICIPANTS: One thousand six hundred seventy-eight patients diagnosed with group D or E retinoblastoma from January 2006 through May 2016. METHODS: Chart review was performed. The patients were divided into primary enucleation and eye-preserving groups, and they were followed up for survival status. The impact of initial treatment on survival was evaluated by Cox analyses. MAIN OUTCOME MEASURES: Overall survival and final eye preservation. RESULTS: After a median follow-up of 43.9 months, 196 patients (12%) died, and the 5-year overall survival was 86%. In total, the eyeball preservation rate was 48%. In this cohort, 1172 patients (70%) had unilateral retinoblastoma, whereas 506 patients (30%) had bilateral disease. For patients with unilateral disease, 570 eyes (49%) underwent primary enucleation, and 602 patients (51%) received eye-preserving therapies initially. During the follow-up (median, 45.6 months), 59 patients (10%) from the primary enucleation group and 56 patients (9.3%) from the eye-preserving group died. Multivariate Cox analyses indicated no significant difference in overall survival between the 2 groups (hazard ratio [HR], 1.25; 95% confidence interval [CI], 0.85-1.84; P = 0.250). For patients with bilateral disease, 95 eyes (19%) underwent primary enucleation, and 411 patients (81%) received eye-preserving therapies initially. During the follow-up (median, 40.1 months), 12 patients (13%) from the primary enucleation group and 69 patients (17%) from the eye-preserving group died. For bilateral retinoblastoma with the worse eye classified as group E, patients undergoing primary enucleation exhibited better overall survival (HR, 2.35; 95% CI, 1.10-5.01; P = 0.027); however, this survival advantage was not evident until passing 22.6 months after initial diagnosis. CONCLUSIONS: Eye-preserving therapies have been used widely for advanced retinoblastoma in China. Patients with bilateral disease whose worse eye was classified as group E and who initially underwent eye-preserving therapies exhibited a worse overall survival. The choice of primary treatment for advanced retinoblastoma should be weighed carefully.

A fluorescent screening method for optimization of conotoxin expression in Pichia pastoris.

Conotoxins are small cysteine-rich peptides secreted by the Conus venom glands, which act on ion channels or membrane receptors with high specificity and potency. Conotoxins are invaluable sources for neuroscience research and drug leads, but their application is hindered by the limited successes in quantitative engineering using either chemical or biotechnological approaches. Here, we explore the Pichia pastoris to express 23 selected conopeptides using a GFP-based fluorescence screen. We found that, in a protease-deficient strain PichiaPink™ Strain 4 (ade2 prb1 pep4), most of the recombinant conopeptides were expressed as two major folding variants including a compact form that was somehow resistant to reduction and high temperature. The GFP-αTxIA was the only one displaying a single band that showed a dose-dependent neurotoxicity on larvae of the insect Plutella xylostella, with a 48-h LD50 lower than 1.12 pmol mg-1 body weight. Furthermore, the recombinant αTxIA after cleavage from the fusion was able to inhibit cell proliferation of the LYCT and HEK293T cell lines with an appearance IC50 of 341 ± 8 and 235 ± 15 nM, respectively. This screening method is straightforward and easy to scale up, providing a versatile tool for further optimization of conotoxin production in the yeast cell.

Magnetic-assisted laparoscopic liver transplantation in swine.

BACKGROUND: Although laparoscopic technology has achieved rapid development in the surgical field, it has not been applied to liver transplantation, primarily because of difficulties associated with laparoscopic vascular anastomosis. In this study, we introduced a new magnetic-assisted vascular anastomosis technique and explored its application in laparoscopic liver transplantation in pigs. METHODS: Two sets of magnetic vascular anastomosis rings (MVARs) with different diameters were developed. One set was used for anastomosis of the suprahepatic vena cava (SHVC) and the other set was used for anastomosis of the infrahepatic vena cava (IHVC) and portal vein (PV). Six laparoscopic orthotopic liver transplantations were performed in pigs. Donor liver was obtained via open surgery. Hepatectomy was performed in the recipients through laparoscopic surgery. Anastomosis of the SHVC was performed using hand-assisted magnetic anastomosis, and the anastomosis of the IHVC and PV was performed by magnetic anastomosis with or without hand assistance. RESULTS: Liver transplants were successfully performed in five of the six cases. Postoperative ultrasonographic examination showed that the portal inflow was smooth. However, PV bending and blood flow obstruction occurred in one case because the MVARs were attached to each other. The durations of loading of MVAR in the laparoscope group and manual assistance group for IHVC and PV were 13 ± 5 vs. 5 ± 1 min (P < 0.01) and 10 ± 2 vs. 4 ± 1 min (P < 0.05), respectively. The durations of MVAR anastomosis in the laparoscope group and manual assistance group for IHVC and PV were 5 ± 1 vs. 1 ± 1 min (P < 0.01), and 5 ± 1 vs. 1 ± 1 min (P < 0.01), respectively. The anhepatic phase was 43 ± 4 min in the laparoscope group and 23 ± 2 min in the manual assistance group (P < 0.01). CONCLUSIONS: Our study showed that magnetic-assisted laparoscopic liver transplantation can be successfully carried out in pigs.

Impacts of psychological contract fulfillment on work attitudes and behaviors during the COVID-19 pandemic: mediating role of perceived organizational support.

The paradigm shifts in HRM during the outbreak of COVID-19 involve new challenges for organizations, whereas it remains unclear how psychological contract fulfillment works on employees' attitudes and behaviors in the new working settings from organizations. This paper explores the impacts of psychological contract fulfillment on employees' work attitudes and behaviors (work engagement, intrinsic motivation, and affective commitment) during the COVID-19 outbreak, and examines the mediating mechanism of perceived organizational support between psychological contract fulfillment and these work-related variables. For the research, a cross-sectional research design and quantitative analysis were adopted. Data were collected via survey questionnaires and from 405 respondents working remotely during the COVID-19 outbreak. The findings revealed that psychological contract fulfillment positively impacted employees' work engagement, intrinsic motivation, and affective commitment during the COVID-19 outbreak, and indicated that perceived organizational support significantly but partially mediates the positive associations between PCF and these work-related variables.

Variations of tongue coating microbiota in children with Henoch-Schönlein purpura nephritis.

BACKGROUND: Variations in the oral microbiota have been significantly correlated with the progress of autoimmune diseases, such as immunoglobulin A nephropathy and Henoch-Schönlein purpura (HSP). However, there is no report outlining the character of tongue coating microbiota variations in children with Henoch-Schönlein purpura nephritis (HSPN). METHOD: A total of 20 children with HSPN and 14 healthy controls were recruited for this research. Tongue coating samples of two groups were collected for 16S rRNA gene sequencing. The diversity, principal component analysis (PCA), nonmetric multidimensional scaling (NMDS), partial least squares discriminant analysis (PLS-DA), and linear discriminant analysis (LDA) effect size (LEfSe) were performed. Microbial function was assessed using the PICRUST. RESULTS: The ACE and Chao indices were greatly lower in the HSPN group than in the HG (P = 0.001). The Shannon and Simpson indices were dramatically reduced in children with HSPN compared with those in the healthy controls (P = 0.005). Bacteroidales, Selenomonadales, Lactobacillales, Fusobacteriales, Neisseriales, and Actinomycetales composed more than 80% of all sequences, while Bacteroidales was the most generous order in both groups. PCA, NMDS and PLS-DA showed a marked difference between the control and HSPN groups. LEfSe analysis showed alteration of tongue coating microbiota in the HSPN group. There were 30 metabolic functions significantly differed between the two groups. CONCLUSIONS: Children with HSPN have substantially various tongue coating microbiota compared to healthy controls. Even though this research does not indicate causality, it is beneficial to enhance the possibility for coming microbial-based treatments to enhance the clinical effects of HSPN in children.

TINCR inhibits the proliferation and invasion of laryngeal squamous cell carcinoma by regulating miR-210/BTG2.

BACKGROUND: Terminal differentiation-induced ncRNA (TINCR) plays an essential role in epidermal differentiation and is involved in the development of various cancers. METHODS: qPCR was used to detect the expression level of TINCR in tissues and cell lines of laryngeal squamous cell carcinoma (LSCC). The potential targets of TINCR were predicted by the bioinformation website. The expression of miR-210 and BTG2 genes were detected by qPCR, and the protein levels of BTG2 and Ki-67 were evaluated by western blot. CCK-8 assay, scratch test, and transwell chamber were used to evaluate the proliferation, invasion, and metastasis ability of LSCC cells. The relationships among TINCR, miR-210, and BTG2 were investigated by bioinformatics software and luciferase reporter assay. The in vivo function of TINCR was accessed on survival rate and tumor growth in nude mice. RESULTS: We used qRT-PCR to detect the expression of TINCR in laryngeal squamous cell carcinoma (LSCC) tissues and cells and found significantly lower levels in cancer tissues compared with adjacent tissues. Additionally, patients with high TINCR expression had a better prognosis. TINCR overexpression was observed to inhibit the proliferation and invasion of LSCC cells. TINCR was shown to exert its antiproliferation and invasion effects by adsorbing miR-210, which significantly promoted the proliferation and invasion of laryngeal squamous cells. Overexpression of miR-210 was determined to reverse the tumour-suppressive effects of TINCR. BTG2 (anti-proliferation factor 2) was identified as the target gene of miR-210, and BTG2 overexpression inhibited the proliferation and invasion of LSCC cells. BTG2 knockdown relieved the inhibitory effects of TINCR on the proliferation and invasion of LSCC. Finally, TINCR upregulation slowed xenograft tumour growth in nude mice and significantly increased survival compared with control mice. CONCLUSION: The results of this study suggest that TINCR inhibits the proliferation and invasion of LSCC by regulating the miR-210/BTG2 pathway, participates in cell cycle regulation, and may become a target for the treatment of LSCC.

Astrocytic Modulation of Supraoptic Oxytocin Neuronal Activity in Rat Dams with Pup-Deprivation at Different Stages of Lactation.

Appropriate interactions between astrocytes and oxytocin neurons in the hypothalamo- neurohypophysial system are essential for normal lactation. To further explore the mechanisms underlying astrocytic modulation of oxytocin neuronal activity, we observed astrocytic plasticity in the supraoptic nucleus of lactating rats with intermittent pup-deprivation (PD, 20 h/day) at early (day 1-5) and middle (day 8-12) stages of lactation. PD at both stages decreased suckling duration and litter's body weight gain. They also significantly increased the expression of glial fibrillary acidic protein (GFAP) in Western blots while increased GFAP filaments and the colocalization of GFAP filaments with aquaporin 4 (AQP4) puncta in astrocyte processes surrounding oxytocin neuronal somata in immunohistochemistry in the supraoptic nucleus. Suckling between adjacent milk ejections but not shortly after them decreased molecular association between GFAP and AQP4. In hypothalamic slices from male rats, oxytocin treatment (0.1 nmol/L, 10 min) significantly reduced the length of GFAP filaments and AQP4 puncta in the processes but increased GFAP staining in the somata. These oxytocin effects were blocked by pretreatment of the slices with N-(1,3,4-Thiadiazolyl) nicotinamide (TGN-020, inhibitor of AQP4, 10 µmol/L, 5 min before oxytocin). In addition, inhibition of AQP4 with TGN-020 blocked excitation in oxytocin neurons evoked by prostaglandin E2, a downstream signal of oxytocin receptor and mediator of oxytocin-evoked burst firing, in whole-cell patch-clamp recordings. These results indicate that AQP4-associated astrocytic plasticity is essential for normal oxytocin neuronal activity during lactation and that PD-evoked hypogalactia is associated with astrocytic process expansion following increased GFAP and AQP4 expressions.

The Implementation and Evaluation of Individual Preference in Robot Facial Expression Based on Emotion Estimation Using Biological Signals.

Recently, robot services have been widely applied in many fields. To provide optimum service, it is essential to maintain good acceptance of the robot for more effective interaction with users. Previously, we attempted to implement facial expressions by synchronizing an estimated human emotion on the face of a robot. The results revealed that the robot could present different perceptions according to individual preferences. In this study, we considered individual differences to improve the acceptance of the robot by changing the robot's expression according to the emotion of its interacting partner. The emotion was estimated using biological signals, and the robot changed its expression according to three conditions: synchronized with the estimated emotion, inversely synchronized, and a funny expression. During the experiment, the participants provided feedback regarding the robot's expression by choosing whether they "like" or "dislike" the expression. We investigated individual differences in the acceptance of the robot expression using the Semantic Differential scale method. In addition, logistic regression was used to create a classification model by considering individual differences based on the biological data and feedback from each participant. We found that the robot expression based on inverse synchronization when the participants felt a negative emotion could result in impression differences among individuals. Then, the robot's expression was determined based on the classification model, and the Semantic Differential scale on the impression of the robot was compared with the three conditions. Overall, we found that the participants were most accepting when the robot expression was calculated using the proposed personalized method.

Residue behavior and safety evaluation of pymetrozine in tea.

BACKGROUND: Pymetrozine is a widely used pesticide. It is challenging to analyze and difficult to manage due to the large gap in its global maximum residue limits (MRLs) in tea. The development of a high-efficiency detection method for the evaluation of the transfer of residual pymetrozine from tea plantations to tea cups is therefore of prime significance. RESULTS: An analytical method for the determination of pymetrozine residues in tea was established based on Cleanert PCX solid-phase extraction. The average recoveries were 72.2-93.7%, with relative standard deviations (RSDs) of less than 12%. The limits of quantification (LOQs) were 0.005 mg·kg-1 in fresh tea leaves and dry tea, and 0.00025 mg·L-1 in tea brew. Pymetrozine degraded rapidly in tea plants with a half-life (t1/2 ) of 1.9 days in open tea plantations, and decreased by 9.4-23.7% in the green tea-processing procedure, which was concentration dependent. The residual pymetrozine levels in green tea collected at 6 and 21 days were below the MRLs in China and EU at a dosage of 30 g a.i. ha-1 , respectively. The leaching rates of pymetrozine from dry tea to tea brew were 58.7-96.3%. Hazard quotient (HQ) values of pymetrozine were significantly <100% when tea shoots were plucked in 6 days, which indicated a negligible risk to humans. CONCLUSION: This work allows the determination of residual pymetrozine in tea and illustrates a low intake risk with the use of pymetrozine in tea plantations. It could serve as reference for further regulation consideration for maximum residue limits (MRLs). © 2020 Society of Chemical Industry.

DNA hypomethylation promotes invasion and metastasis of gastric cancer cells by regulating the binding of SP1 to the CDCA3 promoter.

BACKGROUND: Cell division cycle associated protein-3 (CDCA3) has been reported frequently upregulated in various cancers. It has been progressively realized that changed DNA methylations occur in diverse carcinomas. However, the concrete involvement of CDCA3 and DNA methylation in gastric cancer (GC) still needs to be further elucidated. METHODS: In this study, quantitative reverse-transcription polymerase chain reaction (PCR) was utilized to determine the relative expressions of CDCA3 in GC and normal tissue samples. The methylation condition of CDCA3 was determined by bisulfite-sequencing PCR (BSP) and methylation-specific PCR (MSP). A chromatin immunoprecipitation (ChIP) assay and luciferase activity assay was used for the interaction between transcription factors and promoters and binding site determination, respectively. The effects of knockdown or overexpression of specificity protein 1 (SP1) or CDCA3 on GC cells in vitro were further assessed via wound healing assay, colony formation assay, and matrigel invasion assay. RESULTS: In comparison to paired normal tissues, CDCA3 expressions were significantly increased in the GC tissues. The CDCA3 expression was regulated by DNA methylation, with the CpG island hypomethylation responsible for CDCA3 upregulation of GC. ChIP assays verified that the activity of SP1 binding to the CDCA3 promoter was dramatically increased. When the CDCA3 expression was downregulated in MKN45 cells by knockdown SP1, the proliferation ability, healing ability, and invasive ability were significantly suppressed. CONCLUSION: The process by which SP1 bound to the nearest promoter region was expedited in GC cells, by which DNA was hypomethylated and CDCA3 expression was promoted. The effect on cell proliferation and invasion by CDCA3 was under the regulation of SP1 and also affected by hypomethylation of DNA.