Cloning of Acetyl-CoA Carboxylase accD Subunit Gene from Brassica campestris and Effect of Its Expression on Oil Content

[Objective] This study aimed to clone accD gene from Brassica campestris （Yunnan small rapeseed） with different oil content and analyze the effect of accD gene expression level on rapeseed oil content. [Method] The accD gene encoding β-CT Ⅱsubunit of acetyl-CoA carboxylase was cloned from Yunnan small rapeseeds with different oil content. Then, sequence alignment and the influence upon seed oil content by the expression of accD gene were analyzed. [Result] There were a few base pair substitutes among the accD genes from different Yunnan small rapeseeds, and their homology was up to 98.6%. The expression of accD gene was increased with reproduction development, and reached peak in late-stage siliques. The seed oil content was positively influenced by the expression of accD gene in middle-stage and late-stage siliques. [Conclusion] This study provides a certain theoretical basis for illustrating the molecular mechanism about rapeseed oil content and breeding new high-oil rapeseed cultivars.

Research on the effects of PIAS3 expression on the invasion of glioma TJ905 cells

Objective To investigate the function and possible mechanisms of PIAS3 expression on the invasion of TJ905 cells. Methods PIAS3 overexpression vectors were constructed and PIAS3 siRNA were chemically synthesized, which were separately transfected into TJ905 cells for upregulation or

Impact of KRAS mutation and PTEN expression on cetuximab-treated colorectal cancer

AIM:To investigate the prognostic value of KRAS mutation,and phosphatase and tensin (PTEN) expression in Chinese metastatic colorectal cancer metastatic colorectal cancer (mCRC) patients treated with cetuximab.METHODS:Ninety Chinese mCRC patients treated with cetuximab were evaluated for KRAS mutation and PTEN protein expression by DNA sequencing of codons 12 and 13 and immunohistochemistry,respectively.We then selected 61 patients treated with cetuximab,either in combination with chemotherapy,or alone as a second-line or third-line regimen to assess whether KRAS mutation or PTEN protein expression is associated with the response and the survival time of mCRC patients treated with cetuximab.RESULTS:KRAS mutation was found in 30 (33.3%) tumor samples from the 90 patients,and positive PTEN expression was detected in 58 (64.4%) of the 90 patients.Among the 61 patients who were treated with cetuximab as a second-line or third-line regimen,the resistance to cetuximab was found in 22 patients with KRAS mutation and in 39 patients without KRAS mutation,with a response rate of 4.5% and 46.1% respectively (P=0.001),a shorter median progression-free survival (PFS) time of 14 ± 1.3 wk and 32 ± 2.5 wk respectively (P < 0.001),a median overall survival (OS) time of 11 ± 1.2 mo and 19 ± 1.8 mo respectively (P < 0.001),as well as in 24 patients with negative PTEN expression and in 37 patients with positive PTEN expression respectively (P < 0.001),with a responsive rate of 4.2% and 48.6% respectively,a shorter median PFS survival time of 17 ± 2.0 wk and 28 ± 1.9 wk respectively (P=0.07),and a median OS time of 11 ± 1.3 mo and 18 ± 1.9 mo respectively (P=0.004).Combined KRAS mutation and PTEN expression analysis showed that the PFS and OS time of patients with two favorable prognostic factors were longer than those of patients with one favorable prognostic factor or no favorable prognostic factor (P < 0.001).CONCLUSION:KRAS mutation and PTEN protein expression are significantly correlated with the response rate and survival time of Chinese mCRC patients treated with cetuximab.

LAG-3 expression on tumor-infiltrating T cells in soft tissue sarcoma correlates with poor survival

Objective: To elucidate the role and prognostic significance of lymphocyte activation-gene-3(LAG-3) in soft tissue sarcoma(STS).Methods: The expression of LAG-3 in patient and matched normal blood samples was analyzed by flow cytometry. The localization and prognostic values of LAG-3^+ cells in 163 STS patients were analyzed by immunohistochemistry. In addition, the expression of tumor-infiltrating CD3^+ T, CD4^+ T, and CD8^+ T cells and their role in the prognosis of STS were evaluated by immunohistochemistry. The effect of LAG-3 blockade was evaluated in an immunocompetent MCA205 fibrosarcoma mouse model.Results: Peripheral CD8^+ and CD4^+ T cells from STS patients expressed higher levels of LAG-3 than those from healthy donors.LAG-3 expression in STS was significantly associated with a poor clinical outcome(P = 0.038) and was correlated with high pathological grade(P < 0.001), advanced tumor stage(P = 0.016). Additionally, LAG-3 expression was highly correlated with CD8^+ T-cell infiltration(r = 0.7034, P < 0.001). LAG-3 was expressed in murine tumor-infiltrating lymphocytes, and its blockade decreased tumor growth and enhanced secretion of interferon-gamma by CD8^+ and CD4^+ T cells.Conclusions: LAG-3 blockade may be a promising strategy to improve the effects of targeted therapy in STS.

Influence of FasL gene expression on hepatic metastasis of colorectal carcinoma

BACKGROUND: FasL expression was reported to be asso- ciated with hepatic metastasis of colorectal cancer. The aim of this study was to study FasL gene expression in colorectal carcinoma and its influences on biological behavior and he- patic metastasis of colorectal carcinoma. METHODS: FasL gene expressions were examined with re- verse transcriptase-polymerase chain reaction (RT-PCR) in the primary focus of colorectal carcinoma, adjacent can- cerous mucosae, and metastasized liver focus from colorec- tal cancer. HR-8348 cells of human rectal cancer cell line were transfected with FasL cDNA. Cell growth suppression rate and response to 5-FU and carboplatin were observed and analyzed with the MTT method. RESULTS: FasL gene expression was detected in the prima- ry focus of colorectal cancer ( n = 58), adjacent cancerous mucosae ( n = 58), and metastasized hepatic tumor tissues (n =28). The positive rate of FasL expression was 24% (14/ 58), 8% (5/58), and 100% (58/58) in the primary focus, adjacent cancerous mucosae and metastasized hepatic tumor tissues respectively. FasL expression rate in the me- tastasized hepatic tumor tissues was higher than that in the primary focus (χ2 = 43.49, P<0. 01) and adjacent cance- rous mucosae (χ2=57.66, P<0.01). In a group of patients with hepatic metastasis, the FasL expression rate in primary focus was higher than that in patients without hepatic me- tastasis (χ2=3.96, P <0.05). In vitro study positive expres- sion of FasL was shown in transfected HR-8348 cells. When 5-FU or carboplatin was added, there was a significant difference in growth suppression rate between FasL positive and controlled cancer cells (t=9.02, t = 11.93, P<0.01). Under the same concentration of chemotherapeutic agents, the survival rate of FasL positive HR-8348 cells was higher than that of controlled cells. CONCLUSIONS: FasL positive cancer cells are powerfullyresistant to chemotherapeutic agents. The expression of the FasL gene in colorectal cancer cells is related to immune evasion to escape from being killed by immune cells, show- ing stronger drug-resistance, and it facilitates hepatic me- tastasis.

Effects of Suppressing OsCRY1a Gene Expression on Rice Agronomic Traits

Using primers designed according to the published sequence of rice OsCRY1a gene, we obtained part of the gene fragment by PCR and constructed an RNA interference expression vector with it. To down-regulate the expression level of the gene or lead to the loss-of-function of the gene, the vector was then introduced into rice via Agrobacterium-mediated transformation. Based on the performance of the transgenic plants, the functions of the gene were analyzed and deduced. The results indicated that suppressing the expression of the gene retarded flowering for 16 d in rice with the plant height and grain length significantly increasing whereas other important agronomic traits observed remained unchanged apparently.

Effect of TGIF1 expression on epithelial cadherin and Twist1 protein expression in breast cancer cells

Objective: To observe the effect of the expression of transforming growth factor β-homologous inducible factor 1 (TGIF1) on the expression of epithelial cadherin (E-cadherin) and human Twist-related protein 1 (Twist1) in breast cancer cells. Methods: Total of twenty-four 6-week-old female SPF Balb/c mice were randomly divided into a control group, a TGIF1-silencing group, a TGIF1-normal group, and a TGIF1-overexpression group. In the TGIF1-silencing group, 4T1 breast cancer cells were interfered by lentivirus shRNA (H) lentiviral particles (sc-36659-v) to construct a breast cancer model. TGIF1-normal group used breast cancer cells (4T1) to construct a mouse model of breast cancer. And the TGIF1-overexpression group used 4T1 breast cancer cells with TGIF1 overexpression to establish a mouse model of breast cancer. Determination of TGIF1, E-cadherin and Twist1 protein levels in breast tumor tissue of mice in each group. Results: The tumor volume of mice in the TGIF1-overexpression group was significantly larger than that in the TGIF1-normal group and the TGIF1-silencing group, and the differences between the groups were statistically significant (P <0.05).The expression levels of TGIF1 and Twist1 protein in TGIF1-normal group, TGIF1-silencing group, and TGIF1-overexpression group were significantly higher than those in control group, and E-cadherin was significantly lower than that in control group. The differences between groups were statistically significant (P <0.05).The expression level of TGIF1 and Twist1 protein in TGIF1-silencing group was significantly lower than that in TGIF1-normal group, and E-cadherin was significantly higher than that in TGIF1-normal group (P < 0.05).The expression levels of TGIF1 and Twist1 proteins in the TGIF1-overexpression group were significantly higher than those in the TGIF1-normal group, and E-cadherin was significantly lower than that in the TGIF1-normal group. The differences between the groups were statistically significant (P <0.05). Pearson correlation analysis showed that the expression level of TGIF1 in breast cancer tissue was significantly negatively correlated with the expression level of E-cadherin protein, and was significantly positively correlated with the level of Twist1 protein (P <0.05). Conclusion: TGIF1 can affect the metastasis and invasion of breast cancer by regulating E-cadherin and Twist1 to interfere with the EMT pathway, which deserves further study.

Heficobacter pylori Biofilm Formation and Gene Expression on Abiotic Surfaces Using a Cyanobacterial Extract

The effects of a cyanobacterial extract （CE） on Helicobacter pylori biofilm formation onto hydrophobic and hydrophilic abiotic surfaces and the expression of luxS, flaA, omp18, lpxD and ureA genes associated to biofilm were studied. NCTC11638 reference strain and HP796, a resistant clinical isolate, were grown in Mueller-Hinton broth supplemented with 5% fetal calf serum （FCS） or 1% CE. The ability to form biofilm, viability, morphological changes and gene expression of adhered H. pylori cells were determined. The strains were able to form biofilm on both surfaces with the nutritional supplements analyzed. H. pylori conserved a characteristic bacillary morphology and viability with CE. Cells attachment was higher with CE than FCS regardless of strains and surfaces. The most remarkable increase in gene expression was observed with the ompl8 gene using the CE supplement, indicating the important participation of outer membrane proteins in biofilm establishment. The clinical isolate showed similar and even greater gene expression than the reference strain. The results obtained indicated that the nutrients provided by CE favored biofilm formation with retained pathogenicity that under certain conditions can occur in natural aquatic environments.

Inducible Expression on Multi-epitope of Porcine Circovirus Type 2 and Its Immunological Competence

In order to obtain induction expressed porcine circovirus type 2（PCV2）multi-epitope with good immunogenicity in vitro.Major epitopes of PCV2 were screened in the test,epitopes were composed sequentially,the c DNA sequences were artificially synthesized.Bam HⅠand SalⅠwere directionally cloned into prokaryotic expression vector PEGX-4T-1 multiple cloning site,then BL21 competent cells were transformed,positive clones were screened,IPTG inducible expression was conducted.Expression on target gene was analyzed by SDS-PAGE,fusion protein polypeptide was extracted and purified,immunocompetence of the expressed multi-epitope protein was identified by Westernblot.BALB/c mouse was immuned by fusion protein polypeptide,the antibody was determined by ELISA,immunogenicity was evaluated.Results showed that expression recombinant plasmid pEGX-4T-1-ep contained with seven PCV2 antigen epitopes had been constructed successfully.SDS-PAGE analysis showed that fusion protein polypeptide was expressed effectively in Escherichia coli（E.coli）,and the molecular weight was about 35ku,which existed in the form of solubility.Results of Westernblot showed that the extraction and purification of fusion protein polypeptide and PCV2 positive serum had good reactogenicity.Results of ELISA showed that the purified fusion protein polypeptide could stimulate the body to produce PCV2 specific antibody which had good immunogenicity.Results indicated that the constructed PCV2 multi-epitope had good expression characteristics in vitro,and the expression protein had good immunogenicity.The study provided a basic for the study on PCV2 epitope screening,functional identification and multi-epitope vaccine.

Astrocytic endothelin-1 overexpression impairs learning and memory ability in ischemic stroke via altered hippocampal neurogenesis and lipid metabolism

Vascular etiology is the second most prevalent cause of cognitive impairment globally.Endothelin-1,which is produced and secreted by endothelial cells and astrocytes,is implicated in the pathogenesis of stroke.However,the way in which changes in astrocytic endothelin-1 lead to poststroke cognitive deficits following transient middle cerebral artery occlusion is not well understood.Here,using mice in which astrocytic endothelin-1 was overexpressed,we found that the selective overexpression of endothelin-1 by astrocytic cells led to ischemic stroke-related dementia(1 hour of ischemia;7 days,28 days,or 3 months of reperfusion).We also revealed that astrocytic endothelin-1 overexpression contributed to the role of neural stem cell proliferation but impaired neurogenesis in the dentate gyrus of the hippocampus after middle cerebral artery occlusion.Comprehensive proteome profiles and western blot analysis confirmed that levels of glial fibrillary acidic protein and peroxiredoxin 6,which were differentially expressed in the brain,were significantly increased in mice with astrocytic endothelin-1 overexpression in comparison with wild-type mice 28 days after ischemic stroke.Moreover,the levels of the enriched differentially expressed proteins were closely related to lipid metabolism,as indicated by Kyoto Encyclopedia of Genes and Genomes pathway analysis.Liquid chromatography-mass spectrometry nontargeted metabolite profiling of brain tissues showed that astrocytic endothelin-1 overexpression altered lipid metabolism products such as glycerol phosphatidylcholine,sphingomyelin,and phosphatidic acid.Overall,this study demonstrates that astrocytic endothelin-1 overexpression can impair hippocampal neurogenesis and that it is correlated with lipid metabolism in poststroke cognitive dysfunction.

Effects of antioxidant‑rich Lactiplantibacillus plantarum inoculated alfalfa silage on rumen fermentation, antioxidant and immunity status, and mammary gland gene expression in dairy goats

Background Milk synthesis in lactating animals demands high energy metabolism,which results in an increased production of reactive oxygen metabolites(ROM)causing an imbalance between oxidants and antioxidants thereby inducing oxidative stress(OS)on the animals.To mitigate OS and postpartum disorders in dairy goats and gain insight into the impact of dietary choices on redox status during lactation,a feeding trial was conducted using alfalfa silage inoculated with a high-antioxidant strain of Lactiplantibacillus plantarum.Methods Twenty-four Guanzhong dairy goats(38.1±1.20 kg)were randomly assigned to two dietary treatments:one containing silage inoculated with L.plantarum MTD/1(RSMTD-1),and the other containing silage inoculated with high antioxidant activity L.plantarum 24-7(ES24-7).Results ES24-7-inoculated silage exhibited better fermentation quality and antioxidant activity compared to RSMTD-1.The ES24-7 diet elevated the total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glutathione peroxi-dase(GSH-Px),and catalase(CAT)activities in milk,serum,and feces of lactating goats(with the exception of T-AOC in milk).Additionally,the diet containing ES24-7 inoculated silage enhanced casein yield,milk free fatty acid(FFA)content,and vitamin A level in the goats’milk.Furthermore,an increase of immunoglobulin(Ig)A,IgG,IgM,inter-leukin(IL)-4,and IL-10 concentrations were observed,coupled with a reduction in IL-1β,IL-2,IL-6,interferon(IFN)-γ,and tumor necrosis factor(TNF)-αconcentrations in the serum of lactating goats fed ES24-7.Higher concentrations of total volatile fatty acid(VFA),acetate,and propionate were observed in the rumen fluid of dairy goats fed ES24-7 inoculated silage.Moreover,the diet containing ES24-7 inoculated silage significantly upregulated the expression of nuclear factor erythroid 2 like 2(NFE2L2),beta-carotene oxygenase 1(BCO1),SOD1,SOD2,SOD3,GPX2,CAT,glu-tathione-disulfide reductase(GSR),and heme oxygenase 1(HMOX1)genes in the mammary gland,while decreased the levels of NADPH oxidase 4(NOX4),TNF,and interferon gamma(IFNG).Conclusions These findings indicated that feeding L.plantarum 24-7 inoculated alfalfa silage not only improved rumen fermentation and milk quality in lactating dairy goats but also boosted their immunity and antioxidant status by modulating the expression of several genes related to antioxidant and inflammation in the mammary gland.

Effects of pyraclostrobin on growth,oxidative stress,and gene expression in relation to stress and ATP-binding cassette transporters in Tetrahymena thermophila

Pyraclostrobin(PYR),a widely used fungicide,has negative effects on fish and algae,but its toxicity in protozoa remains unclear.In this study,the effects of PYR on the growth,oxidative stress,and gene expression related to stress and ATP-binding cassette(ABC)transporters in Tetrahymena thermophila were investigated.The result showed that the 96-h IC_(50)of PYR against T.thermophila was 17.2 mg/L.Moreover,PYR inhibited the growth of T.thermophila in concentration-or time-dependent manner.A morphological study revealed that the shape and size of T.thermophila changed,and damage of cell membrane surface was observed by scanning electron microscopy after 96 h of PYR exposure.The activities of superoxide dismutase(SOD)and catalase(CAT)increased throughout the experiment.In contrast,the glutathione(GSH)content was increased at 24 h and 48 h of exposure and decreased at 96 h.Moreover,a significant increase in malondialdehyde(MDA)level was observed in T.thermophila after96 h of exposure.Furthermore,PYR upregulated the HSP703,HSP705,GPx2,and ABAC15 gene expression in the 0.1–5-mg/L groups and downregulated the HSP704,HSP90,TGR,and ABCC52 mRNA levels at 96 h of exposure.These results suggest that PYR may exert adverse effects on T.thermophila by inducing oxidative stress and changing the gene expression related to ABC transporters and stress,which may enrich the understanding of the toxicity mechanism of PYR in aquatic organisms and provide reference data for aquatic ecological risk assessments.

Genome wide investigation of Hsf gene family in Phoebe bournei:identification,evolution,and expression after abiotic stresses

Heat shock transcription factors(Hsfs)have important roles during plant growth and development and responses to abiotic stresses.The identification and func-tion of Hsf genes have been thoroughly studied in various herbaceous plant species,but not woody species,especially Phoebe bournei,an endangered,unique species in China.In this study,17 members of the Hsf gene family were identi-fied from P.bournei using bioinformatic methods.Phyloge-netic analysis indicated that PbHsf genes were grouped into three subfamilies:A,B,and C.Conserved motifs,three-dimensional structure,and physicochemical properties of the PbHsf proteins were also analyzed.The structure of the PbHsf genes varied in the number of exons and introns.Pre-diction of cis-acting elements in the promoter region indi-cated that PbHsf genes are likely involved in responses to plant hormones and stresses.A collinearity analysis dem-onstrated that expansions of the PbHsf gene family mainly take place via segmental duplication.The expression levels of PbHsf genes varied across different plant tissues.On the basis of the expression profiles of five representative PbHsf genes during heat,cold,salt,and drought stress,PbHsf pro-teins seem to have multiple functions depending on the type of abiotic stress.This systematic,genome-wide investigation of PbHsf genes in P.bournei and their expression patterns provides valuable insights and information for further func-tional dissection of Hsf proteins in this endangered,unique species.

Molecular mechanism of hypoxia and alpha-ketoglutaric acid on collagen expression in scleral fibroblasts

AIM:To investigate the molecular mechanisms underlying the influence of hypoxia and alpha-ketoglutaric acid(α-KG)on scleral collagen expression.METHODS:Meta-analysis and clinical statistics were used to prove the changes in choroidal thickness(ChT)during myopia.The establishment of a hypoxic myopia model(HYP)for rabbit scleral fibroblasts through hypoxic culture and the effects of hypoxia andα-KG on collagen expression were demonstrated by Sirius red staining.Transcriptome analysis was used to verify the genes and pathways that hypoxia andα-KG affect collagen expression.Finally,real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used for reverse verification.RESULTS:Meta-analysis results aligned with clinical statistics,revealing a thinning of ChT,leading to scleral hypoxia.Sirius red staining indicated lower collagen expression in the HYP group and higher collagen expression in the HYP+α-KG group,showed that hypoxia reduced collagen expression in scleral fibroblasts,whileα-KG can elevated collagen expression under HYP conditions.Transcriptome analysis unveiled the related genes and signaling pathways of hypoxia andα-KG affect scleral collagen expression and the results were verified by RT-qPCR.CONCLUSION:The potential molecular mechanisms through which hypoxia andα-KG influencing myopia is unraveled and three novel genes TLCD4,TBC1D4,and EPHX3 are identified.These findings provide a new perspective on the prevention and treatment of myopia via regulating collagen expression.

A Facial Expression Recognition Method Integrating Uncertainty Estimation and Active Learning

The effectiveness of facial expression recognition(FER)algorithms hinges on the model’s quality and the availability of a substantial amount of labeled expression data.However,labeling large datasets demands significant human,time,and financial resources.Although active learning methods have mitigated the dependency on extensive labeled data,a cold-start problem persists in small to medium-sized expression recognition datasets.This issue arises because the initial labeled data often fails to represent the full spectrum of facial expression characteristics.This paper introduces an active learning approach that integrates uncertainty estimation,aiming to improve the precision of facial expression recognition regardless of dataset scale variations.The method is divided into two primary phases.First,the model undergoes self-supervised pre-training using contrastive learning and uncertainty estimation to bolster its feature extraction capabilities.Second,the model is fine-tuned using the prior knowledge obtained from the pre-training phase to significantly improve recognition accuracy.In the pretraining phase,the model employs contrastive learning to extract fundamental feature representations from the complete unlabeled dataset.These features are then weighted through a self-attention mechanism with rank regularization.Subsequently,data from the low-weighted set is relabeled to further refine the model’s feature extraction ability.The pre-trained model is then utilized in active learning to select and label information-rich samples more efficiently.Experimental results demonstrate that the proposed method significantly outperforms existing approaches,achieving an improvement in recognition accuracy of 5.09%and 3.82%over the best existing active learning methods,Margin,and Least Confidence methods,respectively,and a 1.61%improvement compared to the conventional segmented active learning method.

Genome-wide identification and expression profiling of photosystem II(PsbX)gene family in upland cotton(Gossypium hirsutum L.)

Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein PsbX has been identified in PSII,which is associated with the oxygen-evolving complex.The expression of PsbX gene protein is regulated by light.PsbX’s central role involves the regulation of PSII,facilitating the binding of quinone molecules to the Qb(PsbA)site,and it additionally plays a crucial role in optimizing the efficiency of photosynthesis.Despite these insights,a comprehensive understanding of the PsbX gene’s functions has remained elusive.Results In this study,we identified ten PsbX genes in Gossypium hirsutum L.The phylogenetic analysis results showed that 40 genes from nine species were classified into one clade.The resulting sequence logos exhibited substantial conservation across the N and C terminals at multiple sites among all Gossypium species.Furthermore,the ortholo-gous/paralogous,Ka/Ks ratio revealed that cotton PsbX genes subjected to positive as well as purifying selection pressure might lead to limited divergence,which resulted in the whole genome and segmental duplication.The expression patterns of GhPsbX genes exhibited variations across specific tissues,as indicated by the analysis.Moreover,the expression of GhPsbX genes could potentially be regulated in response to salt,intense light,and drought stresses.Therefore,GhPsbX genes may play a significant role in the modulation of photosynthesis under adverse abiotic conditions.Conclusion We examined the structure and function of PsbX gene family very first by using comparative genom-ics and systems biology approaches in cotton.It seems that PsbX gene family plays a vital role during the growth and development of cotton under stress conditions.Collectively,the results of this study provide basic information to unveil the molecular and physiological function of PsbX genes of cotton plants.

Global-local combined features to detect pain intensity from facial expression images with attention mechanism

The estimation of pain intensity is critical for medical diagnosis and treatment of patients.With the development of image monitoring technology and artificial intelligence,automatic pain assessment based on facial expression and behavioral analysis shows a potential value in clinical applications.This paper reports a framework of convolutional neural network with global and local attention mechanism(GLA-CNN)for the effective detection of pain intensity at four-level thresholds using facial expression images.GLA-CNN includes two modules,namely global attention network(GANet)and local attention network(LANet).LANet is responsible for extracting representative local patch features of faces,while GANet extracts whole facial features to compensate for the ignored correlative features between patches.In the end,the global correlational and local subtle features are fused for the final estimation of pain intensity.Experiments under the UNBC-McMaster Shoulder Pain database demonstrate that GLA-CNN outperforms other state-of-the-art methods.Additionally,a visualization analysis is conducted to present the feature map of GLA-CNN,intuitively showing that it can extract not only local pain features but also global correlative facial ones.Our study demonstrates that pain assessment based on facial expression is a non-invasive and feasible method,and can be employed as an auxiliary pain assessment tool in clinical practice.

E2E-MFERC:AMulti-Face Expression Recognition Model for Group Emotion Assessment

In smart classrooms, conducting multi-face expression recognition based on existing hardware devices to assessstudents’ group emotions can provide educators with more comprehensive and intuitive classroom effect analysis,thereby continuouslypromotingthe improvementof teaching quality.However,most existingmulti-face expressionrecognition methods adopt a multi-stage approach, with an overall complex process, poor real-time performance,and insufficient generalization ability. In addition, the existing facial expression datasets are mostly single faceimages, which are of low quality and lack specificity, also restricting the development of this research. This paperaims to propose an end-to-end high-performance multi-face expression recognition algorithm model suitable forsmart classrooms, construct a high-quality multi-face expression dataset to support algorithm research, and applythe model to group emotion assessment to expand its application value. To this end, we propose an end-to-endmulti-face expression recognition algorithm model for smart classrooms (E2E-MFERC). In order to provide highqualityand highly targeted data support for model research, we constructed a multi-face expression dataset inreal classrooms (MFED), containing 2,385 images and a total of 18,712 expression labels, collected from smartclassrooms. In constructing E2E-MFERC, by introducing Re-parameterization visual geometry group (RepVGG)block and symmetric positive definite convolution (SPD-Conv) modules to enhance representational capability;combined with the cross stage partial network fusion module optimized by attention mechanism (C2f_Attention),it strengthens the ability to extract key information;adopts asymptotic feature pyramid network (AFPN) featurefusion tailored to classroomscenes and optimizes the head prediction output size;achieves high-performance endto-end multi-face expression detection. Finally, we apply the model to smart classroom group emotion assessmentand provide design references for classroom effect analysis evaluation metrics. Experiments based on MFED showthat the mAP and F1-score of E2E-MFERC on classroom evaluation data reach 83.6% and 0.77, respectively,improving the mAP of same-scale You Only Look Once version 5 (YOLOv5) and You Only Look Once version8 (YOLOv8) by 6.8% and 2.5%, respectively, and the F1-score by 0.06 and 0.04, respectively. E2E-MFERC modelhas obvious advantages in both detection speed and accuracy, which can meet the practical needs of real-timemulti-face expression analysis in classrooms, and serve the application of teaching effect assessment very well.

Genome-Wide Identification of Tomato (Solanum lycopersicum L.) CKX Gene Family and Expression Analysis in the Callus Tissue under Zeatin Treatment

The cytokinin oxidase/dehydrogenase(CKX)enzyme is essential for controlling thefluctuating levels of endogen-ous cytokinin(CK)and has a significant impact on different aspects of plant growth and development.Nonethe-less,there is limited knowledge about CKX genes in tomato(Solanum lycopersicum L.).Here we performed genome-wide identification and analysis of nine SlCKX family members in tomatoes using bioinformatics tools.The results revealed that nine SlCKX genes were unevenly distributed onfive chromosomes(Chr.1,Chr.4,Chr.8,Chr.10,and Chr.12).The amino acid length,isoelectric points,and molecular weight of the nine SlCKX proteins ranged from 453 to 553,5.77 to 8.59,and 51.661 to 62.494 kD,respectively.Subcellular localization analysis indi-cated that SlCKX2 proteins were located in both the vacuole and cytoplasmic matrix;SlCKX3 and SlCKX5 pro-teins were located in the vacuole;and SlCKX1,4,6,7,8,and 9 proteins were located in the cytoplasmic matrix.Furthermore,we observed differences in the gene structures and phylogenetic relationships of SlCKX proteins among different members.SlCKX1-9 were positioned on two out of the three branches of the CKX phylogenetic tree in the multispecies phylogenetic tree construction,revealing their strong conservation within phylogenetic subgroups.Unique patterns of expression of CKX genes were noticed in callus cultures exposed to varying con-centrations of exogenous ZT,suggesting their roles in specific developmental and physiological functions in the regeneration system.These results may facilitate subsequent functional analysis of SlCKX genes and provide valu-able insights for establishing an efficient regeneration system for tomatoes.

Expression Recognition Method Based on Convolutional Neural Network and Capsule Neural Network

Convolutional neural networks struggle to accurately handle changes in angles and twists in the direction of images,which affects their ability to recognize patterns based on internal feature levels. In contrast, CapsNet overcomesthese limitations by vectorizing information through increased directionality and magnitude, ensuring that spatialinformation is not overlooked. Therefore, this study proposes a novel expression recognition technique calledCAPSULE-VGG, which combines the strengths of CapsNet and convolutional neural networks. By refining andintegrating features extracted by a convolutional neural network before introducing theminto CapsNet, ourmodelenhances facial recognition capabilities. Compared to traditional neural network models, our approach offersfaster training pace, improved convergence speed, and higher accuracy rates approaching stability. Experimentalresults demonstrate that our method achieves recognition rates of 74.14% for the FER2013 expression dataset and99.85% for the CK+ expression dataset. By contrasting these findings with those obtained using conventionalexpression recognition techniques and incorporating CapsNet’s advantages, we effectively address issues associatedwith convolutional neural networks while increasing expression identification accuracy.