Genome-Wide Identification of TLP Gene Family in Populus trichocarpa and Functional Characterization of PtTLP6, Preferentially Expressed in Phloem.

Thaumatin-like proteins (TLPs) in plants are involved in diverse biotic and abiotic stresses, including antifungal activity, low temperature, drought, and high salinity. However, the roles of the TLP genes are rarely reported in early flowering. Here, the TLP gene family was identified in P. trichocarpa. The 49 PtTLP genes were classified into 10 clusters, and gene structures, conserved motifs, and expression patterns were analyzed in these PtTLP genes. Among 49 PtTLP genes, the PtTLP6 transcription level is preferentially high in stems, and GUS staining signals were mainly detected in the phloem tissues of the PtTLP6pro::GUS transgenic poplars. We generated transgenic Arabidopsis plants overexpressing the PtTLP6 gene, and its overexpression lines showed early flowering phenotypes. However, the expression levels of main flowering regulating genes were not significantly altered in these PtTLP6-overexpressing plants. Our data further showed that overexpression of the PtTLP6 gene led to a reactive oxygen species (ROS) burst in Arabidopsis, which might advance the development process of transgenic plants. In addition, subcellular localization of PtTLP6-fused green fluorescent protein (GFP) was in peroxisome, as suggested by tobacco leaf transient transformation. Overall, this work provides a comprehensive analysis of the TLP gene family in Populus and an insight into the role of TLPs in woody plants.

Phomopsterone B Alleviates Liver Fibrosis through mTOR-Mediated Autophagy and Apoptosis Pathway.

Liver fibrosis is the initial pathological process of many chronic liver diseases. Targeting hepatic stellate cell (HSC) activation is an available strategy for the therapy of liver fibrosis. We aimed to explore the anti-liver fibrosis activity and potential mechanism of phomopsterone B (PB) in human HSCs. The results showed that PB effectively attenuated the proliferation of TGF-ß1-stimulated LX-2 cells in a concentration-dependent manner at doses of 1, 2, and 4 µM. Quantitative real-time PCR and Western blot assays displayed that PB significantly reduced the expression levels of α-SMA and collagen I/III. AO/EB and Hoechst33342 staining and flow cytometry assays exhibited that PB promoted the cells' apoptosis. Meanwhile, PB diminished the number of autophagic vesicles and vacuolated structures, and the LC3B fluorescent spots indicated that PB could effectively inhibit the accretion of autophagosomes in LX-2 cells. Moreover, rapamycin and MHY1485 were utilized to further investigate the effect of mTOR in autophagy and apoptosis. The results demonstrated that PB regulated autophagy and apoptosis via the mTOR-dependent pathway in LX-2 cells. In summary, this is the first evidence that PB effectively alleviates liver fibrosis in TGF-ß1-stimulated LX-2 cells, and PB may be a promising candidate for the prevention of liver fibrosis.

Correlation analysis between vitamin A, D, and E status with altitude, seasonal variation, and other factors, among children aged 0-6 years in a Chinese population living in the Tibetan plateau of Ganzi prefecture.

OBJECTIVE: We attempted to understand the status of vitamin (Vit) A, D, and E in children aged 0-6 living in the Tibetan plateau areas of Ganzi prefecture, to provide the basis for relevant government departments to carry out physical examinations of these children and to prevent and cure four key diseases (Infantile diarrhea, nutritional anemia, rickets, and infantile pneumonia). METHODS: Serum retinol and tocopherol levels were detected using high-performance liquid chromatography (HPLC). Serum levels of 25-(OH)D were detected by high-performance liquid chromatography-tandem mass spectrometry (LC-MS). The polynomial logistic regression was used to analyze the effects of age, season, altitude, and gender on Vit A, D, and E levels. RESULTS: Vit A and D had the lowest mean serum levels before the age of 1 year and with the most significant deficiency rates. The lowest Vit E levels were seen in the Toddlerhood group. The rates of deficiency and insufficiency were the highest. Vit A, D, and E levels were significantly affected by seasonal changes and were significantly higher in the summer than in any other season. Vit A and D were significantly affected by altitude, and their levels were lowest above 4 km. CONCLUSION: The overall levels of Vit A, D, and E in children aged 0-6 in the Tibetan plateau areas of Ganzi prefecture were lower than those in the plain's areas.

Untargeted metabolomics analysis by gas chromatography/time-of-flight mass spectrometry of human serum from methamphetamine abusers.

Methamphetamine (METH) abuse has become a global public health problem. However, the potential mechanisms involving METH-induced metabolic disorders have thus far remained poorly understood. Metabolomics can provide a clue for the cause of apparent changes and consequently be used to investigate the METH-induced dysregulation of metabolite expression and the mechanism of metabolic disorder mechanism. This laboratory investigation included 80 METH abusers and 80 healthy people. The serum metabolites were detected and analysed by gas chromatography/time-of-flight mass spectrometry. Raw data were processed with the software MS DIAL, which includes deconvolution, peak alignment and compound identification. The data matrix was processed by univariate and multivariate analyses for significant metabolite screening with the criteria of variable importance in projection values > 1, fold change > 1.5 and the t test (p value < 0.05). Significant differences in 16 metabolites (deoxycholic acid, cholic acid, hydroxylamine, etc.) in serum were found between the METH abuse group and the control group. Energy metabolic pathways and several amino acid metabolic pathways (alanine, aspartic acid and glutamate metabolism and tryptophan metabolism) were primarily involved. Further analysis indicated that the area under the receiver operating characteristic curve (AUC) was 0.998 for these 16 metabolites. Among the metabolites, three carbohydrates (d-ribose, cellobiose and maltotriose) had an AUC of 0.975, which were determined as potential markers of abuse. We observed metabolic disturbances in METH abusers, particularly perturbation in energy metabolism and amino acid metabolism, which can provide new insights into the search for biomarkers and the mechanisms underlying the adverse effects of METH on human health.

Opposite estrogen effects of estrone and 2-hydroxyestrone on MCF-7 sensitivity to the cytotoxic action of cell growth, oxidative stress and inflammation activity.

There are many kinds of estrogens, and endogenous estrogens produce a variety of estrogen metabolites with similar structure but with different physiological effects after metabolism in vivo. Studies have shown that estrone (E1) widely occurs in the environment and animal-derived food. Because of its estrogen effect, E1 can have adverse effects on the human body as an endocrine disruptor. In this study, we found that E1 and 2-hydroxyestrone (2-OH-E1), the hydroxylation metabolite of estrogen, have opposite proliferative effects on breast cancer cells (MCF-7) through cell proliferation experiments and comparison of their effects by molecular docking and detection of ROS, Ca2+, and cell pathway proteins. The effects of 2-methoxyestrone (2-MeO-E1) and 16α-hydroxyestrone (16α-OH-E1) on the biochemical and protein levels of MCF-7 were further studied to compare the effects of metabolic sites and modes on estrogen effects. Hydroxylation of E1 at the C2 site weakened the estrogen effect, down-regulated the expression of the mammalian target of rapamycin (mTOR) and protein kinase B (Akt) pathway proteins, inhibited the proliferation of cancer cells, and enhanced anti-oxidative stress and anti-inflammation. Methoxylation at the C2 position also inhibited the expression of inflammatory and oxidative stress pathway proteins but did not greatly affect the estrogen effects. However, hydroxylation on C16 had no significant effect on the biological effects of estrogen. Therefore, the structural changes of estrogen on C2 are important reasons for the different physiological effects of estrogen and its metabolites. Thus, by regulating the gene Cytochrome P450 1B1(CYP1B1), which affects the hydroxylation metabolism of estrogen, and promoting the hydroxylation of estrone at the C2 position, the estrogen effect of estrone can be effectively reduced, thus reducing the harm its poses in food and the environment.

Endoscopic thyroidectomy using gasless axillary approach for low-risk papillary thyroid carcinoma.

To evaluate the application of endoscopic thyroidectomy using gasless axillary approach (ET-GA) for low-risk papillary thyroid carcinoma (PTC). Patients with T1N0M0 Ⅰ PTC undergoing unilateral thyroid lobectomy with central neck dissection in Taizhou Cancer Hospital during January 2019 to June 2021 were enrolled in the study, including 35 cases treated with ET-GA (ET-GA group) and 35 cases treated with conventional open thyroidectomy (COT group). The surgical treatment effect, cosmetic effect and the effect on neck function were compared between two groups. Neck function was evaluated by neck pain score, neck injury index and dysphagia index. Cosmetic effect was evaluated by cosmetic effect satisfaction score. In the ET-GA group, all unilateral thyroid lobectomy with central neck dissection were successfully completed, and no case was converted to open surgery. The number of central lymph nodes dissected in the ET-GA was not statistically different from that in the COT group (>0.05), but the operation time was longer than that of the COT group (<0.01). In the ET-GA group, 2 cases (5.7%) had transient vocal cord paralysis and 1 case (2.9%) had postoperative bleeding. In the COT group, 1 case (2.9%) had transient vocal cord paralysis, no postoperative bleeding. There was no significant difference in the complication rate between two groups (>0.05). At and postoperatively, there was no significant difference in neck pain score and neck injury index between two groups (both >0.05); dysphagia index was lower in ET-GA group, but the difference was not statistically significant (>0.05). The cosmetic effect satisfaction score of ET-GA group was higher than that in the COT group at postoperatively (4.3±0.6 vs.1.0, <0.01). ET-GA has the same efficacy and safety as conventional open thyroidectomy in the treatment of low-risk PTC, and it improves the satisfaction of postoperative cosmetics.

Advances in targeted therapy for anaplastic thyroid carcinoma.

Anaplastic thyroid carcinoma (ATC) is a highly malignant and aggressive thyroid malignancy with rapid onset and poor prognosis. There is no effective treatment for ATC yet. Molecular targeted therapy provides a new idea for ATC treatment. Tyrosine kinase inhibitor lenvatinib has potential in treating ATC patients with favorable efficacy in clinical trials. The effectiveness of the v-raf murine sarcoma viral oncogene homologue B1 () gene inhibitor dabrafenib in combination with trametinib for the treatment of positive ATC patients has been demonstrated in clinical trials. The has proposed dabrafenib in combination with trametinib as the preferred modality for the treatment of patients with positive ATC. The immune checkpoint inhibitor pembrolizumab can be applied to treat thyroid cancer with high tumor mutational load and may be considered as the preferred modality for the treatment of ATC patients with high programmed death ligand-1 expression. The mammalian target of rapamycin pathway inhibitors, peroxisome proliferators-activated receptor γ agonists, endothelial growth factor receptors-targeting monoclonal antibody cetuximab and novel vascular blocker fosbretabulin are still in the clinical research stage, which are expected to provide new directions for the development of novel targeted drugs. This article reviews the current research progress on targeted drugs for the treatment of ATC.

Two new withanolides from Physalis minima L.

Two new withanolides named physaminilides L (1) and M (2), together with four known ones (3-6) were isolated from the Physalis minima L. The structures were established by analysis of the HR ESIMS, IR and NMR spectroscopic data. The absolute configurations were determined through NOESY and ECD spectra. For compounds 1-5 assayed at 20 µM and compound 6 at 10 µM, inhibition rates of hepatic fibrosis were 22.19%, 15.29%, 37.07%, 9.27%, 12.45%, and 37.03%, respectively.

Withanolides from the active extract of Physalis angulate and their anti-hepatic fibrosis effects.

ETHNOPHARMACOLOGICAL RELEVANCE: Physalis angulata L., a traditional Chinese medicine called "Kuzhi" in China, was used traditionally to treat liver diseases (eg. icterus, hepatitis) as well as malaria, asthma, and rheumatism. AIM OF THE STUDY: Our study aimed to investigate the withanolides with anti-hepatic fibrosis effect from P. angulate. MATERIALS AND METHODS: Withanolides were obtained from the EtOH extract of P. angulate by bioassay-molecular networking analysis-guided isolation using column chromatography and normal/reversed-phase semipreparative HPLC. The structures of new withanolides were elucidated by combinations of spectroscopic techniques with NMR and ECD calculations. MTT cell viability assay, AO/EB staining method, cell wound healing assay, ELISA and Western blot experiments were employed to evaluate the anti-hepatic fibrosis activity and to uncover related mechanism. Molecular docking analysis and cellular thermal shift assay were used to evaluate and verify the interaction between the active withanolides and their potential targets. RESULTS: Eight unreported withanolides, withagulides A-H (1-8), along with twenty-eight known ones were obtained from P. angulate. Withanolides 6, 9, 10, 24, 27, and 29-32 showed marked anti-hepatic fibrosis effect with COL1A1 expression inhibition above 50 %. Physalin F (9), the main component in the active fraction, significantly decreased the TGF ß1-stimulated expressions of collagen I and α-SMA in LX-2 cells. Mechanism study revealed that physalin F exerted its anti-hepatic fibrosis effect via the PI3K/AKT/mTOR signaling pathway. CONCLUSION: This study suggested that withanolides were an important class of natural products with marked anti-hepatic fibrosis effect. The main withanolide physalin F might be a promising candidate for hepatic fibrosis treatment. The work provided experimental foundation for the use of P. angulate to treat hepatic fibrosis.

Oral vaccination of BacFish-vp6 against grass carp reovirus evoking antibody response in grass carp.

To obtain a vaccine and evaluate its immune efficacy as an oral vaccine against Grass Carp Reovirus (GCRV), a donor vector pFastBac-FA-VP6-ph-VP6 containing two GCRV vp6 genes, one driven by the Megalobrama amblycephala ß-actin promoter and another one controlled by baculovirus polyhedrin promoter was constructed to generate the recombinant baculovirus BacFish-vp6. From the hemolymph of 5th instar silkworm inoculated with BacFish-vp6, a 53 kDa recombinant VP6 protein could be detected. And the infected pupae collected at 120 h post-inoculation with BacFish-vp6 were used to make freeze-dried powder as an oral vaccine. When the grass carps were orally administrated with feed containing 1%, 5% and 10% of the freeze-dried powder, their specific antibody against VP6 could be detected. Further studies showed that vp6 has been transcripted in the grass carp kidney (CIK) cells infected with BacFish-vp6 and the different tissues of orally vaccinated fish. All the results suggested the powder of the silkworm pupae infected with BacFish-vp6 could possibly be used as an orally administered vaccine.

Construction of nano-drug delivery and antitumor system of stimuli-responsive polypeptides.

The size of the nanoparticles is moderate and the dispersion is well, which will not be recognized nonspecifically and clearance by the endothelial reticular system. In this study, stimuli-responsive polypeptides nano-delivery system has been constructed, which can realize the response to various stimuli in the tumor microenvironment. Tertiary amine groups are grafted to the side chain of polypeptides as the point of charge reversal and particle expansion. In addition, a new kind of liquid crystal monomer was prepared by substituting cholesterol-cysteamine, which can promote polymers to realize the transformation of spatial conformation by adjusting the ordered arrangement of macromolecules. The introduction of hydrophobic elements greatly enhanced the self-assembly performance of polypeptides, which could effectively improve the drug loading and encapsulation rate of nanoparticles. Nanoparticles could achieve targeted aggregation in tumor tissues, and there were no toxicity and side effects on normal bodies during treatment, with good safety in vivo.

Silencing of AJAP1 expression by promoter methylation activates the Wnt/ß-catenin signaling pathway to promote tumor proliferation and metastasis in salivary adenoid cystic carcinoma.

Background: Salivary adenoid cystic carcinoma (SACC) is a unique malignant tumor of the salivary gland with poor prognosis, which is not effective with chemotherapy and targeted drugs. Therefore, it is important to explore the molecular mechanism underlying SACC invasion and metastasis to develop novel therapeutic strategies and targets in clinical research. Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot (WB) were performed to detect the expression of Adherens Junctions Associated Protein 1 (AJAP1). Methylation-specific PCR was used to evaluate the methylation of the AJAP1 promoter. AJAP1 was overexpressed or knocked down by lentivirus-mediated transfection. Kaplan-Meier analysis was conducted to create a survival curve and the log-rank test was used to analyze the overall survival (OS). The prognostic correlation was assessed using univariate and multivariate Cox regression analyses. Co-immunoprecipitation (Co-IP) was utilized to pull down the possible binding protein of AJAP1 and laser scanning confocal microscopy was applied to detect the subcellular localization of AJAP1, E-cadherin, and ß-catenin. Cell viability, colony formation, wound healing, and Transwell invasion assays were performed to evaluate the function of AJAP1 in vitro. A subcutaneous xenograft assay in nude mice was performed to verify the function of AJAP1 in vivo. Results: AJAP1 was downregulated in SACC tumors and was closely related to SACC lymph node/distant metastasis, which was an independent risk factor for SACC prognosis. Methylation-specific PCR confirmed that high methylation of the AJAP1 promoter was the main cause of its silencing. Overexpression or knockdown of AJAP1 in SACC cells could significantly inhibit or promote the proliferation, invasion, and metastasis of SACC cells, respectively, in both the in vitro and in vivo experiments. Mechanically, we found that AJAP1 binds to E-cadherin and ß-catenin to form a complex in cytomembrane, reducing the nuclear translocation of ß-catenin and blocking the Wingless/Integrated/ß-catenin (Wnt/ß-catenin) signaling pathway to play a suppressive role in cancer. Conclusions: In conclusion, these results suggest that the downregulation of AJAP1 protein expression may play a certain role in progression and metastasis of SACC. Our study indicates that AJAP1 may be a potential prognostic molecular marker and therapeutic target for SACC.

Room temperature phosphorescence sensor for Hg2+ based on Mn-doped ZnS quantum dots.

Mercury pollution is one of the most serious concerns to human health and the environment. The development of highly sensitive and selective sensors to detect toxic mercury ions has been the focus of scientific research. In this study, L-cysteine-capped Mn-doped ZnS quantum dots (QDs) have been synthesized and used for the room temperature phosphorescence detection of Hg2+. The phosphorescence of the Mn-doped ZnS QDs could be selectively quenched in the presence of Hg2+. Under optimal conditions, good linear correlations were obtained over the concentration range from 2.0 x 10(-8) mol/L to 4.5 x 10(-6) mol/L with a detection limit of 3.8 x 10(-9) mol/L. Additionally, the long lifetime of the phosphorescence of the Mn-doped ZnS QDs can avoid the interference of the autofluorescence and scattering light of the background, which facilitates their application in real samples. The possible quenching mechanism was examined by UV-vis absorption spectra and Rayleigh scattering spectra.

The Effects of Saposhnikovia divaricata Aqueous Extracts on the Inflammation and Intestinal Microflora in Allergic Rhinitis Mice.

Background: Allergic rhinitis (AR) is a type I allergic disease induced by IgE. Traditional Chinese medicine Saposhnikovia divaricata (Turcz.) Schischk (SD) has anti-inflammatory and antiallergic effects. Materials and Methods: AR model mice were constructed with ovalbumin (OVA) sensitization to observe the improving effect of SD treatment on AR by counting the number of sneezing and rubbing the nose, hematoxylin-eosin, periodic acid-Schiff, and toluidine blue stains. In addition, the allergy and inflammatory cytokines levels and inflammatory cells were observed by ELISA and Wright's-Giemsa stain. The protein levels of the TLR4/TRAF6/NF-κB and IL-6/ROR-γt/STAT3 pathways were measured by immunohistochemistry, quantitative real-time PCR, and western blot. The intestinal microflora abundance in mice was observed by 16S rDNA high-throughput sequencing. Results: SD treatment inhibited the sneezing and rubbing times of the nose, decreased the degree of a dense arrangement of cells and mucosal swelling and the number of goblet and mast cells of nasal lavage fluid, reduced the levels of IgE, histamine, Leukotriene B4, IL-4, IL-5, TNF-α, IL-6, and IL-17, the eosinophils, neutrophils, and lymphocytes number, the LR4, TRAF6, IL-6, ROR-γt, and STAT3 mRNA levels, respectively, while, it increased the IL-2, IL-10, IFN-γ, and TGF-ß1 proteins. SD treatment inhibited the NF-κB, p-STAT3, TLR4, TRAF6, and p-IκBα/IκBα proteins. Besides, the effects of OVA and SD treatments were significantly correlated with the abundance of intestinal microflora. The abundances of Cytophagales, Burkholderia, Alteromonadales, Lactococcus, and Clostridiaceae were changed in SD treatment on AR mice. Conclusions: This study provides a possibility that the improvement effect of SD treatment on allergies and inflammation in AR mice may be related to the TLR4/TRAF6/NF-κB and IL-6/ROR-γt/STAT3 pathways and intestinal microflora modulation.

Does media sentiment affect stock prices? Evidence from China's STAR market.

Objective: This paper explores the impact of media sentiment on stock prices on the Shanghai Stock Exchange Science and Technology Innovation Board (hereinafter the STAR market) from a behavioral finance perspective. Methods: We collect Baidu News coverage of STAR-listed firms as the text, and measure text sentiment using a machine learning-based text analysis technique. We then empirically examine the impact of media sentiment on STAR market stock prices from two aspects: IPO pricing efficiency and IPO first-day stock performance. Results: (1) Media sentiment has no significant impact on IPO pricing efficiency, thus suggesting that institutional investors participating in such offerings are generally not affected by media sentiment. (2) Optimistic media sentiment has a positive impact on IPO first-day returns, which indicates that individual investors are more easily influenced by media sentiment and therefore likely to abandon their rational judgment. (3) Media sentiment had a greater impact on IPO first-day returns during the COVID-19 pandemic than those before it, which suggests that individual investors are more influenced by media sentiment during pandemics. Discussion: Our findings deepen the understanding of stock price formation on the STAR market, which provide a statistical basis for formulating policy directions and investment strategies.

A Systematic Pan-Cancer Analysis of CASP3 as a Potential Target for Immunotherapy.

CASP3 is the gene encoding caspase-3, a specific protease that cleaves substrates such as poly-ADP ribose polymerase and acetyl-DEVD-7-amino-4-methylcoumarin. This enzymatic activity leads to DNA fragmentation, which is a hallmark of apoptosis. Although recent studies have demonstrated that CASP3 plays a vital role in tumour suppression by promoting apoptosis, these reports did not consider systematic pan-cancer analyses. Therefore, we performed a specific pan-cancer analysis using The Cancer Genome Atlas and Genotype-Tissue Expression databases to analyse CASP3 expression in terms of cancer prognosis, DNA methylation status, tumour mutative burden (TMB), and microsatellite instability (MSI), as well as immune cell infiltration in different tumours and the molecular mechanisms underlying these. We found that CASP3 expression was significantly associated with the prognosis of most tumours. Additionally, promoter methylation status was associated with CASP3 expression in bladder urothelial carcinoma, oesophageal carcinoma, kidney renal clear cell carcinoma, kidney renal papillary cell carcinoma, lung squamous cell carcinoma, prostate adenocarcinoma, sarcoma, testicular germ cell tumours, and uterine corpus endometrial carcinoma. TMB and MSI were associated with CASP3 expression in 15 tumours. Moreover, CASP3 expression was correlated with the tumour microenvironment in nearly all tumour types. Further, we observed that in addition to apoptosis, CASP3 action plausibly involves B cell activation, antigen presentation, immune responses, chemokine receptors, and inflammatory function. Our study thus provides a relatively comprehensive understanding of the carcinogenicity of CASP3 in different tumours and suggests that CASP3 is a potential prognostic marker.

[Dynamics of carbon and nitrogen balance during leaf senescence of maize seedlings induced by low nitrogen stress]. / 低氮诱导玉米幼苗叶片衰老过程中碳氮平衡的动态变化.

Timing of leaf senescence is important to ensure maize yield. In this study, we investigated the dynamics of carbon and nitrogen balance during leaf senescence in two maize inbred lines PH6WC and PH4CV under normal (4 mmol·L-1, CK) and low nitrogen (0.04 mmol·L-1, LN) treatments. Leaf phenotype, photosynthetic characteristics, nitrogen and sugar contents, and carbon to nitrogen ratio of the second and third leaves were analyzed after 2, 4, 6 and 8 days of cultivation. Results showed that leaf size, biomass, relative chlorophyll content, net photosynthetic rate, soluble sugar content, and starch content of the second and third leaves were decreased, while nitrogen production capacity was increased under low nitrogen treatment compared to the control, with the changes of the second leaf being earlier than that of the third leaf. For all the leaf traits, the variation scales of PH6WC were larger than that of PH4CV under low nitrogen stress, and only the C/N ratio in the seedling leaves was significantly increased. In addition, leaf senescence of PH4CV was slower than PH6WC due to its stronger ability in maintaining carbon and nitrogen balance. In conclusion, low nitrogen could induce leaf senescence of maize seedlings. High C/N ratio could promote leaf senescence. There are significant differences in carbon and nitrogen balance ability of seedling leaves between two maize genotypes under low nitrogen stress.

A Novel and Effective Model to Predict Skip Metastasis in Papillary Thyroid Carcinoma Based on a Support Vector Machine.

Introduction: Skip metastasis, referred to as lymph node metastases to the lateral neck compartment without involvement of the central compartment, is generally unpredictable in papillary thyroid carcinoma (PTC). This study aims to establish an effective predictive model for skip metastasis in PTC. Meterials and Methods: Retrospective analysis was performed of clinical samples from 18192 patients diagnosed with thyroid cancer between 2016 to 2020. The First Affiliated Hospital of Wenzhou Medical University. The lateral lymph node metastasis was occureed in the training set (630 PTC patients) and validation set (189 PTC patients). The univariate and multivariate analyses were performed to detect the predictors of skip metastasis and the support vector machine (SVM) was used to establish a model to predict skip metastasis. Results: The rate of skip metastasis was 13.3% (84/631). Tumor size (≤10 mm), upper location, Hashimoto's thyroiditis, extrathyroidal extension, absence of BRAFV600E mutation, and less number of central lymph node dissection were considered as independent predictors of skip metastasis in PTC. For the training set, these predictors performed with 91.7% accuracy, 86.4% sensitivity, 92.2% specificity, 45.2% positive predictive value (PPV), and 98.9% negative predictive value (NPV) in the model. Meanwhile, these predictors showed 91.5% accuracy,71.4% sensitivity, 93.1% specificity, 45.5% PPV, and 97.6% NPV in validation set. Conclusion: This study screened the predictors of the skip lateral lymph node metastasis and to establish an effective and economic predictive model for skip metastasis in PTC. The model can accurately distinguish the skip metastasis in PTC using a simple and affordable method, which may have potential for daily clinical application in the future.

Acceptance and Online Interpretation of "Gender-Neutral Pronouns": Performance Asymmetry by Chinese English as a Foreign Language Learners.

The present study (N = 109) set out to examine the role of cross-linguistic differences as a source of potential difficulty in the acceptance and online interpretation of the English singular they by Chinese English as a Foreign Language (EFL) learners across two levels of second-language proficiency. Experiment 1 operationalized performance through an untimed acceptability judgment test and Experiment 2 through a self-paced reading task. Statistical analyses yielded an asymmetric pattern of results. Experiment 1 indicated that unlike native English speakers who generally accepted the singular they with all antecedent types, two Chinese EFL groups consisting of English majors (higher level) and non-English majors (lower level) both rated it as the least acceptable pronoun regardless of their proficiency level. In contrast, Experiment 2 demonstrated that like native English speakers, both Chinese EFL groups were not disrupted in their reading by the use of the singular they most of the time, although its online interpretation was modulated by L2 proficiency levels of the participants. While the English majors were not affected by the use of the singular they, the non-English majors spent a significantly longer time reading the latter region of the sentences where the singular they was used. In short, the results of the two experiments seem to indicate that under no time constraint, L2 speakers showed a heightened degree of grammar sensitivity, whereas when there was a time constraint, their grammatical sensitivity was reduced by a greater need to focus on meaning. The difficulty for Chinese EFL learners to acquire the singular they may be located at the restructuring of their existing knowledge of the plural feature of they [-PLURAL] in their mental lexicon and the adaptation to the sociocultural norms of the target language. The pedagogical implications of the findings are discussed.

Unreduced Male Gamete Formation in Cymbidium and Its Use for Developing Sexual Polyploid Cultivars.

Polyploidy plays an important role in crop improvement. Polyploid plants, particularly those produced through unreduced gametes (2n gametes), show increased organ size, improved buffering capacity for deleterious mutations, and enhanced heterozygosity and heterosis. Induced polyploidy has been widely used for improving floriculture crops, however, there are few reported sexual polyploid plants in the floriculture industry. This study evaluated nine cultivars of Cymbidium Swartz and discovered that 2n male gametes occurred in this important orchid. Depending on cultivars, 2n male gamete formation frequencies varied from 0.15 to 4.03%. Interspecific hybrids generally produced more 2n male gametes than traditional cultivars. To generate sexual polyploid plants, seven pairs of crosses were made, which produced five triploid and two tetraploid hybrids. Two triploid hybrids were evaluated for in vitro regeneration and growth characteristics. Compared to the diploid parents, the triploids were more easily regenerated through rhizomes or protocorms, and regenerated plants had improved survival rates after transplanting to the greenhouse. Furthermore, the sexual polyploid plants had more compact growth style, produced fragrant flowers, and demonstrated heterosis in plant growth. Through this study, a reliable protocol for selection of appropriate parents for 2n gamete production, ploidy level evaluation, in vitro culture of polyploid progenies, and development of new polyploid cultivars was established. Our study with Cymbidium suggests that the use of 2n gametes is a viable approach for improving floriculture crops.