Identifying and validating MMP family members(MMP2,MMP9,MMP12,and MMP16)as therapeutic targets and biomarkers in kidney renal clear cell carcinoma(KIRC)

Kidney Renal Clear Cell Carcinoma(KIRC)is a malignant tumor that carries a substantial risk of morbidity and mortality.The MMP family assumes a crucial role in tumor invasion and metastasis.This study aimed to uncover the mechanistic relevance of the MMP gene family as a therapeutic target and diagnostic biomarker in Kidney Renal Clear Cell Carcinoma(KIRC)through a comprehensive approach encompassing both computational and molecular analyses.STRING,Cytoscape,UALCAN,GEPIA,OncoDB,HPA,cBioPortal,GSEA,TIMER,ENCORI,DrugBank,targeted bisulfite sequencing(bisulfite-seq),conventional PCR,Sanger sequencing,and RT-qPCR based analyses were used in the present study to analyze MMP gene family members to accurately determine a few hub genes that can be utilized as both therapeutic targets and diagnostic biomarkers for KIRC.By performing STRING and Cytohubba analyses of the 24 MMP gene family members,MMP2(matrix metallopeptidase 2),MMP9(matrix metallopeptidase 9),MMP12(matrix metallopeptidase 12),and MMP16(matrix metallopeptidase 16)genes were denoted as hub genes having highest degree scores.After analyzing MMP2,MMP9,MMP12,and MMP16 via various TCGA databases and RT-qPCR technique across clinical samples and KIRC cell lines,interestingly,all these hub genes were found significantly overexpressed at mRNA and protein levels in KIRC samples relative to controls.The notable effect of the up-regulated MMP2,MMP9,MMP12,and MMP16 was also documented on the overall survival(OS)of the KIRC patients.Moreover,targeted bisulfite-sequencing(bisulfite-seq)analysis revealed that promoter hypomethylation pattern was associated with up-regulation of hub genes(MMP2,MMP9,MMP12,and MMP16).In addition to this,hub genes were involved in various diverse oncogenic pathways.The MMP gene family members(MMP2,MMP9,MMP12,and MMP16)may serve as therapeutic targets and prognostic biomarkers in KIRC.

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.

Genome-wide identification,characterization and functional prediction of the SRS gene family in sesame(Sesamum indicum L.)

Sesame(Sesamum indicum L.)is an ancient oilseed crop of the Pedaliaceae family with high oil content and potential health benefits.SHI RELATED SEQUENCE(SRS)proteins are the transcription factors(TFs)specific to plants that contain RING-like zinc finger domain and are associated with the regulation of several physiological and biochemical processes.They also play vital roles in plant growth and development such as root formation,leaf development,floral development,hormone biosynthesis,signal transduction,and biotic and abiotic stress responses.Nevertheless,the SRS gene family was not reported in sesame yet.In this study,identification,molecular characterization,phylogenetic relationship,cis-acting regulatory elements,protein-protein interaction,syntenic relationship,duplication events and expression pattern of SRS genes were analyzed in S.indicum.We identified total six SiSRS genes on seven different linkage groups in the S.indicum genome by comparing with the other species,including the model plant Arabidopsis thaliana.The SiSRS genes showed variation in their structure like2–5 exons and 1–4 introns.Like other species,SiSRS proteins also contained‘RING-like zinc finger'and‘LRP1'domains.Then,the SiSRS genes were clustered into subclasses via phylogenetic analysis with proteins of S.indicum,A.thaliana,and some other plant species.The cis-acting regulatory elements analysis revealed that the promoter region of SiSRS4(SIN_1011561)showed the highest 13 and 16 elements for light-and phytohormone-responses whereas,SiSRS1(SIN_1015187)showed the highest 15 elements for stress-response.The ABREs,or ABA-responsive elements,were found in a maximum of 8 copies in the SiSRS3(SIN 1009100).Moreover,the available RNA-seq based expression of SiSRS genes revealed variation in expression patterns between stress-treated and non-treated samples,especially in drought and salinity conditions in.S.indicum.Two SiSRS genes like SiSRS1(SIN_1015187)and SiSRS5(SIN_1021065),also exhibited variable expression patterns between control vs PEG-treated sesame root samples and three SiSRS genes,including SiSRS1(SIN_1015187),SiSRS2(SIN_1003328)and SiSRS5(SIN_1021065)were responsive to salinity treatments.The present outcomes will encourage more research into the gene expression and functionality analysis of SiSRS genes in S.indicum and other related species.

Identification of the lysine and histidine transporter family in Camellia sinensis and the characterizations in nitrogen utilization

In plants,the lysine and histidine transporter(LHT)family represent a class of proteins that mediate the uptake,translocation,and utilization of amino acids.The tea plant(Camellia sinensis)is a perennial evergreen with a relatively high level of amino acids.However,systematic identification and molecular characterization of the LHT gene family has rarely been reported in tea plants.In this study,22 CsLHTs were identified from the‘Shuchazao’genome and classified into two groups.The modeled three-dimensional structure and the conserved domains presented a high similarity among the LHTs proteins.Moreover,it was predicted that a few genes were conserved through the analysis of the physiochemical characters,structures and cis-elements in promoters.The expression patterns in tea plants revealed that CsLHT7 was mainly expressed in the roots,and CsLHT4 and CsLHT11 exhibited relatively high expression in both the roots and leaves.Moreover,the expression of all three genes could be induced by organic nitrogen.Additionally,heterogeneous expression of CsLHT4,CsLHT7 and CsLHT11 in Arabidopsis thaliana decreased the aerial parts biomass compared with that in WT plants while significantly increased the rosette biomass only for CsLHT11transgenic plants versus WT plants.Overall,our results provide fundamental information about CsLHTs and potential genes in N utilization for further analysis in tea plants.

Aldo-keto reductase family member C3(AKR1C3)promotes hepatocellular carcinoma cell growth by producing prostaglandin F2α

Hepatocellular carcinoma(HCC)is a leading cause of death worldwide.Current therapies are effective for HCC patients with early disease,but many patients suffer recurrence after surgery and have a poor response to chemotherapy.Therefore,new therapeutic targets are needed.We analyzed gene expression profiles between HCC tissues and normal adjacent tissues from public databases and found that the expression of genes involved in lipid metabolism was significantly different.The analysis showed that AKR1C3 was upregulated in tumors,and high AKR1C3 expression was associated with a poorer prognosis in HCC patients.In vitro,assays demonstrated that the knockdown of AKR1C3 or the addition of the AKR1C3 inhibitor indomethacin suppressed the growth and colony formation of HCC cell lines.Knockdown of AKR1C3 in Huh7 cells reduced tumor growth in vivo.To explore the mechanism,we performed pathway enrichment analysis,and the results linked the expression of AKR1C3 with prostaglandin F2 alpha(PGF2a)downstream target genes.Suppression of AKR1C3 activity reduced the production of PGF2a,and supplementation with PGF2a restored the growth of indomethacin-treated Huh7 cells.Knockdown of the PGF receptor(PTGFR)and treatment with a PTGFR inhibitor significantly reduced HCC growth.We showed that indomethacin potentiated the sensitivity of Huh7 cells to sorafenib.In summary,our results indicate that AKR1C3 upregulation may promote HCC growth by promoting the production of PGF2α,and suppression of PTGFR limited HCC growth.Therefore,targeting the AKR1C3-PGF2a-PTGFR axis may be a new strategy for the treatment of HCC.

Genome-Wide Exploration of the Grape GLR Gene Family and Differential Responses of VvGLR3.1 and VvGLR3.2 to Low Temperature and Salt Stress

Grapes,one of the oldest tree species globally,are rich in vitamins.However,environmental conditions such as low temperature and soil salinization significantly affect grape yield and quality.The glutamate receptor(GLR)family,comprising highly conserved ligand-gated ion channels,regulates plant growth and development in response to stress.In this study,11 members of the VvGLR gene family in grapes were identified using whole-genome sequence analysis.Bioinformatic methods were employed to analyze the basic physical and chemical properties,phylogenetic trees,conserved domains,motifs,expression patterns,and evolutionary relationships.Phylogenetic and collinear analyses revealed that the VvGLRs were divided into three subgroups,showing the high conservation of the grape GLR family.These members exhibited 2 glutamate receptor binding regions(GABAb and GluR)and 3-4 transmembrane regions(M1,M2,M3,and M4).Real-time quantitative PCR analysis demonstrated the sensitivity of all VvGLRs to low temperature and salt stress.Subsequent localization studies in Nicotiana tabacum verified that VvGLR3.1 and VvGLR3.2 proteins were located on the cell membrane and cell nucleus.Additionally,yeast transformation experiments confirmed the functionality of VvGLR3.1 and VvGLR3.2 in response to low temperature and salt stress.Thesefindings highlight the significant role of the GLR family,a highly conserved group of ion channels,in enhancing grape stress resistance.This study offers new insights into the grape GLR gene family,providing fundamental knowledge for further functional analysis and breeding of stress-resistant grapevines.

MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1-extracellular signal-regulated kinase-1/2 axis

BACKGROUND MicroRNAs(miRNAs)regulate gene expression and play a critical role in cancer physiology.However,there is still a limited understanding of the function and regulatory mechanism of miRNAs in gastric cancer(GC).AIM To investigate the role and molecular mechanism of miRNA-145-5p(miR145-5p)in the progression of GC.METHODS Real-time polymerase chain reaction(RT-PCR)was used to detect miRNA expression in human GC tissues and cells.The ability of cancer cells to migrate and invade was assessed using wound-healing and transwell assays,respectively.Cell proliferation was measured using cell counting kit-8 and colony formation assays,and apoptosis was evaluated using flow cytometry.Expression of the epithelial-mesenchymal transition(EMT)-associated protein was determined by Western blot.Targets of miR-145-5p were predicated using bioinformatics analysis and verified using a dual-luciferase reporter system.Serpin family E member 1(SERPINE1)expression in GC tissues and cells was evaluated using RT-PCR and immunohistochemical staining.The correlation between SERPINE1 expression and overall patient survival was determined using Kaplan-Meier plot analysis.The association between SERPINE1 and GC progression was also tested.A rescue experiment of SERPINE1 overexpression was conducted to verify the relationship between this protein and miR-145-5p.The mechanism by which miR-145-5p influences GC progression was further explored by assessing tumor formation in nude mice.RESULTS GC tissues and cells had reduced miR-145-5p expression and SERPINE1 was identified as a direct target of this miRNA.Overexpression of miR-145-5p was associated with decreased GC cell proliferation,invasion,migration,and EMT,and these effects were reversed by forcing SERPINE1 expression.Kaplan-Meier plot analysis revealed that patients with higher SERPINE1 expression had a shorter survival rate than those with lower SERPINE1 expression.Nude mouse tumorigenesis experiments confirmed that miR-145-5p targets SERPINE1 to regulate extracellular signal-regulated kinase-1/2(ERK1/2).CONCLUSION This study found that miR-145-5p inhibits tumor progression and is expressed in lower amounts in patients with GC.MiR-145-5p was found to affect GC cell proliferation,migration,and invasion by negatively regulating SERPINE1 levels and controlling the ERK1/2 pathway.

Tanshinone ⅡA improves Alzheimer’s disease via RNA nuclearenriched abundant transcript 1/microRNA-291a-3p/member RAS oncogene family Rab22a axis

BACKGROUND Alzheimer’s disease(AD)is a neurodegenerative condition characterized by oxidative stress and neuroinflammation.Tanshinone ⅡA(Tan-ⅡA),a bioactive compound isolated from Salvia miltiorrhiza plants,has shown potential neuroprotective effects;however,the mechanisms underlying such a function remain unclear.AIM To investigate potential Tan-ⅡA neuroprotective effects in AD and to elucidate their underlying mechanisms.METHODS Hematoxylin and eosin staining was utilized to analyze structural brain tissue morphology.To assess changes in oxidative stress and neuroinflammation,we performed enzyme-linked immunosorbent assay and western blotting.Additionally,the effect of Tan-ⅡA on AD cell models was evaluated in vitro using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Genetic changes related to the long non-coding RNA(lncRNA)nuclear-enriched abundant transcript 1(NEAT1)/microRNA(miRNA,miR)-291a-3p/member RAS oncogene family Rab22a axis were assessed through reverse transcription quantitative polymerase chain reaction.RESULTS In vivo,Tan-ⅡA treatment improved neuronal morphology and attenuated oxidative stress and neuroinflammation in the brain tissue of AD mice.In vitro experiments showed that Tan-ⅡA dose-dependently ameliorated the amyloid-beta 1-42-induced reduction of neural stem cell viability,apoptosis,oxidative stress,and neuroinflammation.In this process,the lncRNA NEAT1-a potential therapeutic target-is highly expressed in AD mice and downregulated via Tan-ⅡA treatment.Mechanistically,NEAT1 promotes the transcription and translation of Rab22a via miR-291a-3p,which activates nuclear factor kappa-B(NF-κB)signaling,leading to activation of the pro-apoptotic B-cell lymphoma 2-associated X protein and inhibition of the anti-apoptotic B-cell lymphoma 2 protein,which exacerbates AD.Tan-ⅡA intervention effectively blocked this process by inhibiting the NEAT1/miR-291a-3p/Rab22a axis and NF-κB signaling.CONCLUSION This study demonstrates that Tan-ⅡA exerts neuroprotective effects in AD by modulating the NEAT1/miR-291a-3p/Rab22a/NF-κB signaling pathway,serving as a foundation for the development of innovative approaches for AD therapy.

Therapeutic Communication Methods Targeting Families and Family Members: A Literature Review

Background and Purpose: Therapeutic communication is a new term in family health care nursing, defined by Hohashi (2019) as a method of family intervention, and characterized by inclusion of not only verbal conversation but also nonverbal interaction. However, specific therapeutic communication methods have not been systematized. The purpose of this study was to clarify therapeutic communication methods for families/family members from the perspectives of verbal communication and non-verbal communication through a review of existing literature. Methods: We conducted a search using the medical literature databases PubMed and Ichushi-Web using the keywords “therapeutic communication”. Analysis was performed on seven articles from PubMed and 14 articles from Ichushi-Web that described therapeutic communication methods performed by healthcare professionals for families/family members. Through directed content analysis, therapeutic communication methods were subcategorized, and classified into three categories: verbal communication, non-verbal communication, and verbal/non-verbal communication. Results: A total of 23 subcategories were extracted. Verbal communication included 11 subcategories, such as “asking questions using the communicatee’s words as they are”. Non-verbal communication included five subcategories, such as “noticing changes in the content of the communicatee’s story”. And verbal/non-verbal communication featured seven subcategories, such as “making the communicatee aware of one’s own beliefs”. Conclusion: Therapeutic communication methods included basic care/caring in family interviews/meetings, as well as verbal communication and non-verbal communication that act on family/family members’ beliefs. It is believed that changes in family/family members’ beliefs can be used to eliminate, reduce, or improve problematic conditions in the family. .

The Impact of Adolescent and Young Adult Cancer-AJA-on Family Functioning.Study of Family Relationships in Terms of Adaptability and Cohesion

Objectives:The arrival of cancer in adolescents and young adults(aged 15 to 24 years)-Adolescents and young adults(AJA)-corresponds to a fragile period during which the adulthood of the young person and the evolution of family ties mobilize the family as a whole.Therefore,cancer,beyond its individual traumatic dimension,affects the whole family,which can modify family ties and family functioning.Our objective is to evaluate family functioning from the complex model evaluating cohesion and adaptability when an adolescent or young adult has cancer.Methods:Adolescents and young adults with cancer(n=41),mothers(n=41),and fathers(n=13)participated in this study.They completed the Family Adaptation and Cohesion Scales(FACES Ⅲ)questionnaire.Family functioning when an aya is ill has been compared to that of families without any disease.Results:a comparison of the mean scores of perceived cohesion and adaptability of face Ⅲ indicates no significant difference for cohesion.In contrast,the averages of the adaptability scores of our sample with those of the general population indicate that families with cancer hais generally feel more“adaptable”than the non-clinical population.These results are statistically significant for AJA,but also for mothers and fathers.Regarding the mean scores of ideal cohesion and ideal adaptability,there are no significant differences between fathers in our sample and fathers in the general population.In contrast,mothers in our sample had less ideal adaptability than those in the general population.In aya patients with cancer,the scores of both adaptability and cohesion were significantly different from those of non-diseased adolescents.Conclusion:Cancer leads to many changes in family relationships,making it difficult to empower the young patient and latent the evolution of the relationship.

Functional prediction of tomato PLATZ family members and functional verification of Sl PLATZ17

PLATZ is a novel zinc finger DNA-binding protein that plays an important role in regulating plant growth and development and resisting abiotic stress.However,there has been very little research on the function of this family gene in tomatoes,which limits its application in germplasm resource improvement.Therefore,the PLATZ gene family was identified and analyzed in tomato,and its roles were predicted and verified to provide a basis for in-depth research on SlPLATZ gene function.In this study,the PLATZ family members of tomato were identified in the whole genome,and 19 SlPLATZ genes were obtained.Functional prediction was conducted based on gene and promoter structure analysis and RNA-seq-based expression pattern analysis.SlPLATZ genes that responded significantly under different abiotic stresses or were significantly differentially expressed among multiple tissues were screened as functional gene resources.SlPLATZ17 was selected for functional verification by experiment-based analysis.The results showed that the downregulation of SlPLATZ17 gene expression reduced the drought and salt tolerance of tomato plants.Tomato plants overexpressing SlPLATZ17 had larger flower sizes and long,thin petals,adjacent petals were not connected at the base,and the stamen circumference was smaller.This study contributes to understanding the functions of the SlPLATZ family in tomato and provides a reference for functional gene screening.

Characterization of a Novel Esterase Belonging to Family V from Marinobacter flavimaris

Lipolytic enzymes have attracted enormous attentions because of their ability in ester hydrolysis,ester synthesis,transesterification and other biochemical reactions.Bacteria are important sources of lipolytic enzymes applied in industry.Here,a novel lipolytic enzyme encoded by esterase gene est1347 was identified in Marinobacter flavimaris WLL162,and was purified and characterized.The lipolytic enzyme Est1347 consisted of 312 amino acid residues and a 21-amino-acids N-terminal signal peptide with a predicted molecular weight of 34.2 kDa.It belongs to family V of bacterial lipolytic enzymes based on the amino acid sequence homology analysis.Est1347 is a mesophilic and alkali-resistant enzyme with the highest activity at 45℃and pH 8.5;it is stable at temperatures below 50℃and pH 7.5–11.0.Est1347 showed a preference for middle-length chain substrate p-NPC10 and a wide range of other substrates.The Km,Vmax,Kcat and Kcat/Km values of Est1347 for p-NPC10 in pH 8.5 at 45℃were 0.9411 mmol L^(−1),1285μmol min^(−1)mg^(−1),698.91 s^(−1)and 743.65 s^(−1)(mmol L^(−1))^(−1),respectively.It is also tolerant to the metal ions,organic solvents and detergents.In conclusion,the esterase Est1347 laid a foundation for further study of bacterial lipolytic enzyme family V.

Correlates of Mistimed Pregnancy and Unmet Need for Family Planning among Women of Reproductive Age in Sandema, Ghana

Background: Globally, an estimated 80 million unintended pregnancies comprising both mistimed and unwanted pregnancies are recorded yearly. Yet only half of the women at risk of mistimed pregnancy use contraceptives. In developing countries, over 100 million females have unmet need, and national surveys in Ghana indicate 23% unmet need rate. Methods: Using a cross-sectional community-based approach, a sample size of 300 women of reproductive age were selected using multi-step cluster sampling techniques. The study was quantitative, using structured interviewer-administered questionnaires. Results: Two-third (66%) of the women in reproductive age still had unmet need, 71% were currently pregnant, and more than a third (36%) confirmed ever having a mistimed pregnancy. Fifty-three percent (53%) of the women confirmed never communicating with their partners on family planning issues, a little below half (45%) took their own health care decisions. Seventy nine percent (79%) ever received family planning services from a health professional. Factors related to unmet needs included mistimed pregnancy, level of education, preferred birth/pregnancy interval, communication between partners and the autonomy to spend self-earnings. Conclusion: Considering that high rates of unmet need results in mistimed pregnancy, improved policies around the influence of unmet need on mistimed pregnancies are needed.

Postpartum Family Planning at the Timbuktu Hospital in Mali

Introduction: Postpartum family planning is the prevention of pregnancies during the 12 months following childbirth. Few studies have been devoted to postpartum family planning in Mali. Our work will contribute to reducing unmet need for family planning. Objective: To study the use of contraceptive methods in the postpartum period in the obstetrics and gynecology unit of Timbuktu hospital. Materials and Methods: This was a descriptive and analytical cross-sectional study with prospective collection of data from January 1, 2022 to December 31, 2023. All women who gave birth having chosen and benefited from a contraceptive method were included. The statistical test used was the Fisher test with a significance threshold fixed at 5%. Results: The frequency of contraception in the postpartum period was 17.03%. The average age of clients was 26.14% with extremes of 14 and 45 years. They were paupiparous at 56.4% with an inter-birth interval of less than 12 months at 12.3%. More than half of the counseling (58.5%) was done during postnatal visits. The methods chosen were implants at 48.1%, injectable progestins at 21.3%, intrauterine device at 14.7%, miro-progestin pills at 13.5%, tubal ligation at 1 .4% and condoms at 1%. The regular follow-up rate was 51.1% of cases and 25.6% had no follow-up. Conclusion: The overall rate of postpartum family planning of 17.08% remains low. Improving FP staff skills will reduce unmet needs and contribute to increasing contraceptive prevalence in Timbuktu.

Exponentially Convergent Multiscale Finite Element Method

We provide a concise review of the exponentially convergent multiscale finite element method(ExpMsFEM)for efficient model reduction of PDEs in heterogeneous media without scale separation and in high-frequency wave propagation.The ExpMsFEM is built on the non-overlapped domain decomposition in the classical MsFEM while enriching the approximation space systematically to achieve a nearly exponential convergence rate regarding the number of basis functions.Unlike most generalizations of the MsFEM in the literature,the ExpMsFEM does not rely on any partition of unity functions.In general,it is necessary to use function representations dependent on the right-hand side to break the algebraic Kolmogorov n-width barrier to achieve exponential convergence.Indeed,there are online and offline parts in the function representation provided by the ExpMsFEM.The online part depends on the right-hand side locally and can be computed in parallel efficiently.The offline part contains basis functions that are used in the Galerkin method to assemble the stiffness matrix;they are all independent of the right-hand side,so the stiffness matrix can be used repeatedly in multi-query scenarios.

Identification and Molecular Characterization of the Alkaloid Biosynthesis Gene Family in Dendrobium catenatum

As one of the main active components of Dendrobium catenatum, alkaloids have high medicinal value. The physicochemicalproperties, conserved domains and motifs, phylogenetic analysis, and cis-acting elements of the genefamily members in the alkaloid biosynthesis pathway of D. catenatum were analyzed by bioinformatics, and theexpression of the genes in different years and tissues was analyzed by qRT-PCR. There are 16 gene families,including 25 genes, in the D. catenatum alkaloid biosynthesis pathway. The analysis of conserved domains andmotifs showed that the types, quantities, and orders of domains and motifs were similar among members ofthe same family, but there were significant differences among families. Phylogenetic analysis indicated that thegene family members showed some evolutionary conservation. Cis-acting element analysis revealed that therewere a large number of light-responsive elements and MYB (v-myb avian myeloblastosis viral oncogene homolog)-related elements in these genes. qRT-PCR showed that expressions of gene family members involved in alkaloidsynthesis were different in different years and tissues of D. catenatum. This study provides a theoretical basisfor further exploration of the regulatory mechanisms of these genes in the alkaloid biosynthesis of D. catenatum.

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.

Identification and Analysis of the WRKY Transcription Factor Gene Family in Verbena bonariensis

The WRKY transcription factor gene family is one of the unique gene families in plants.It plays an important role in response to abiotic stresses such as cold and drought,hormone signal transduction,regulation of biosynthesis,leaf senescence seed germination,etc.However,little information is available about WRKY transcription factors in Verbena bonariensis.In this study,70 VbWRKY genes were identified from the whole genome.The phylogenetic analysis of the WRKY gene family in V.bonariensis and Arabidopsis shows that the WRKY genes in V.bonariensis can be divided into three groups:I,II,and III,which contain 13,47,and 10 members,respectively.Group II can be further divided into five subclasses:IIa(5),IIb(10),IIc(18),IId(6),and IIe(8).Conservative motif analysis showed that 64 proteins encoded by the VbWRKY gene had conserved motifs 1,2 and 3,and the same subclass motif elements were approximately the same.The collinearity analysis showed that there were 44 homologous gene pairs among the VbWRKYs,and these homologous gene pairs may have the same function.Promoter sequence analysis showed that the VbWRKY gene has multiple cis-acting elements,including not only cis-acting elements related to low-temperature and light responses,but also cis-acting elements related to hormone regulation,Among them,most VbWRKY genes contain response elements about low-temperature,and 30 VbWRKY genes contain low-temperature response elements(LTR),and 61 VbWRKY genes contain abscisic acid response elements(ABRE),indicating that VbWRKY plays a crucial role in plant growth and abiotic stress.According to the expression of VbWRKY in the cold stress and different tissues transcriptome,70 VbWRKY genes played their respective roles in various tissues and stages to regulate plant growth,Also,some of them participated in the process of cold stress tolerance,52 VbWRKYs showed significant differences in expression under cold stress,and 37 VbWRKY genes were up-regulated under cold stress.9 VbWRKY genes were selected for quantitative real-time PCR(qRT-PCR)analysis under low-temperature stress,and the results showed that all 9 genes were upregulated under low-temperature stress.Ultimately,the present study provides a comprehensive analysis of the predicted V.bonariensis WRKY genes family,which provided a theoretical basis for the study of low-temperature resistance and growth and development of V.bonariensis.

GST family genes in jujube actively respond to phytoplasma infection

Jujube witches’broom(JWB)caused by phytoplasma has a severely negative effect on multiple metabolisms in jujube.The GST gene family in plants participates in the regulation of a variety of biotic and abiotic stresses.This study aims to identify and reveal the changes in the jujube GST gene family in response to phytoplasma infection.Here,70 ZjGSTs were identified in the jujube genome and divided into 8 classes.Among them,the Tau-class,including 44 genes,was the largest.Phylogenetic analysis indicated that Tau-class genes were highly conserved among species,such as Arabidopsis,cotton,chickpea,and rice.Through chromosome location analysis,37.1%of genes were clustered,and 8 of 9 gene clusters were composed of Tau class members.Through RT-PCR,qRT-PCR and enzyme activity detection,the results showed that the expression of half(20/40)of the tested ZjGSTs was inhibited by phytoplasma infection in field and tissue culture conditions,and GST activity was also significantly reduced.In the resistant and susceptible varieties under phytoplasma infection,ZjGSTU49-ZjGSTU54 in the cluster IV showed opposite expression patterns,which may be due to functional divergence during evolution.Some upregulated genes(ZjGSTU45,ZjGSTU49,ZjGSTU59,and ZjGSTU70)might be involved in the process of jujube against JWB.The yeast two-hybrid results showed that all 6 Tauclass proteins tested could form homodimers or heterodimers.Overall,the comprehensive analysis of the jujube GST gene family revealed that ZjGSTs responded actively to phytoplasma infection.Furthermore,some screened genes(ZjGSTU24,ZjGSTU49-52,ZjGSTU70,and ZjDHAR10)will contribute to further functional studies of jujube-phytoplasma interactions.

Product family modeling technology for customized cosmetic packaging design based on basic-element theory

Background:As the market demands change,SMEs(small and medium-sized enterprises)have long faced many design issues,including high costs,lengthy cycles,and insufficient innovation.These issues are especially noticeable in the domain of cosmetic packaging design.Objective:To explore innovative product family modeling methods and configuration design processes to improve the efficiency of enterprise cosmetic packaging design and develop the design for mass customization.Methods:To accomplish this objective,the basic-element theory has been introduced and applied to the design and development system of the product family.Results:By examining the mapping relationships between the demand domain,functional domain,technology domain,and structure domain,four interrelated models have been developed,including the demand model,functional model,technology model,and structure model.Together,these models form the mechanism and methodology of product family modeling,specifically for cosmetic packaging design.Through an analysis of a case study on men’s cosmetic packaging design,the feasibility of the proposed product family modeling technology has been demonstrated in terms of customized cosmetic packaging design,and the design efficiency has been enhanced.Conclusion:The product family modeling technology employs a formalized element as a module configuration design language,permeating throughout the entire development cycle of cosmetic packaging design,thus facilitating a structured and modularized configuration design process for the product family system.The application of the basic-element principle in product family modeling technology contributes to the enrichment of the research field surrounding cosmetic packaging product family configuration design,while also providing valuable methods and references for enterprises aiming to elevate the efficiency of cosmetic packaging design for the mass customization product model.