Recovery from spindle checkpoint-mediated arrest requires a novel Dnt1-dependent APC/C activation mechanism.

The activated spindle assembly checkpoint (SAC) potently inhibits the anaphase-promoting complex/cyclosome (APC/C) to ensure accurate chromosome segregation at anaphase. Early studies have recognized that the SAC should be silenced within minutes to enable rapid APC/C activation and synchronous segregation of chromosomes once all kinetochores are properly attached, but the underlying silencers are still being elucidated. Here, we report that the timely silencing of SAC in fission yeast requires dnt1+, which causes severe thiabendazole (TBZ) sensitivity and increased rate of lagging chromosomes when deleted. The absence of Dnt1 results in prolonged inhibitory binding of mitotic checkpoint complex (MCC) to APC/C and attenuated protein levels of Slp1Cdc20, consequently slows the degradation of cyclin B and securin, and eventually delays anaphase entry in cells released from SAC activation. Interestingly, Dnt1 physically associates with APC/C upon SAC activation. We propose that this association may fend off excessive and prolonged MCC binding to APC/C and help to maintain Slp1Cdc20 stability. This may allow a subset of APC/C to retain activity, which ensures rapid anaphase onset and mitotic exit once SAC is inactivated. Therefore, our study uncovered a new player in dictating the timing and efficacy of APC/C activation, which is actively required for maintaining cell viability upon recovery from the inhibition of APC/C by spindle checkpoint.

Ca9-yMgyNaGd0.667(PO4)7:Eu2+: A Single Eu2+-Doped Full-Spectrum White Light Emission Phosphor with High Color Rendering Index.

The multi-cationic site occupation control of rare earth doped phosphor materials is one of the key factors in achieving single-phase full-spectrum emission. In this work, the site distribution of Eu2+ in Ca9NaGd0.667(PO4)7 (CNGP):x%Eu2+ phosphors with multi-cationic sites was determined. Under 385 nm excitation, CNGP:x%Eu2+ samples exhibited a cyan emission composed of three sub-Gaussian components. The CNGP:1.5%Eu2+ phosphor was structurally modified through the partial substitution of Mg2+ for Ca2+ according to the crystallographic site engineering theory, and corresponding luminescent properties had been adjusted. When the concentration of Mg2+ changed, the Ca9-yMgyNaGd0.667(PO4)7:1.5%Eu2+ (CNGPE:yMg2+) phosphors exhibited a tunable emission from cyan to orange and achieved white light emission at y = 0.6. Based on the change of the crystal field environment, the mechanism of Mg2+ substitution affecting the luminescence characteristics of CNGP:x%Eu2+ phosphors was researched in detail. The light-emitting diode device consisted of a CNGPE:0.6Mg2+ phosphor and a 385 nm chip and had a high color rendering index (CRI = 91.9), low correlated color temperature (CCT = 4852 K), and excellent color stability. The optical performance of this device makes it promising to be used in full spectrum lighting.

B(C6 F5 )3 -Catalyzed [2+3]-Cyclative o,m-diC-H Functionalization of Phenols.

Metal-free catalytic C-H functionalization is highly desired for the construction of C-C bonds. We herein report a highly chemoselective consecutive C-H [2+3]-cyclative functionalization for the simultaneous formation of two C-C bonds with construction of polycyclic phenols catalyzed by commercially available and low-cost B(C6 F5 )3 . This catalytic system tolerates a wide range of substrate scope, providing a series of 2,6,7,8-tetrahydroacenaphthylen-3-ol-type polycyclic compounds efficiently. Several derivatizations of the catalytic products have also been conducted to show the potential application of this method in synthesis of polycyclic compounds.

Development and validation of a predictive model of abnormal uterine bleeding associated with ovulatory dysfunction: a case-control study.

BACKGROUND: Abnormal uterine bleeding associated with ovulatory dysfunction (AUB-O) is a typical gynecological disease that can affect women of various ages. Being able to identify women at risk of AUB-O could allow physicians to take timely action. This study aimed to identify the influencing factors of AUB-O in Chinese women, and then develop and validate a predictive model. METHODS: In this multicenter case-control study, 391 women with AUB-O and 838 controls who came from nine hospitals in Zhejiang province were recruited between April 2019 and January 2022. All the participants completed a structured questionnaire including general characteristics, lifestyle and habits, menstrual and reproductive history, and previous diseases. The predictive model was developed on a group of 822 women and validated on a group of 407 women. Logistic regression was adopted to investigate the influencing factors and develop the model, and validation was then performed. RESULTS: The independent predictive factors of AUB-O were age (OR 1.073, 95% CI 1.046-1.102, P < 0.001), body mass index (OR 1.081, 95% CI 1.016-1.151, P = 0.015), systolic blood pressure (OR 1.016, 95% CI 1.002-1.029, P = 0.023), residence (OR 2.451, 95% CI 1.727-3.478, P < 0.001), plant-based diet (OR 2.306, 95% CI 1.415-3.759, P < 0.001), fruits eating (OR 1.887, 95% CI 1.282-2.776, P = 0.001), daily sleep duration (OR 0.819; 95% CI 0.708-0.946, P = 0.007), multiparous (parity = 1, OR 0.424, 95% CI 0.239-0.752, P = 0.003; parity > 1, OR 0.450, 95% CI 0.247-0.822, P = 0.009), and history of ovarian cyst (OR 1.880, 95% CI 1.305-2.710, P < 0.001). The predictive ability (area under the curve) in the development group was 0.77 (95% CI 0.74-0.81), while in the validation group it was 0.73 (95% CI 0.67-0.79). The calibration curve was in high coincidence with the standard curve in the development group, and similar to the validation group. A tool for AUB-O risk calculation was created. CONCLUSIONS: Nine influencing factors and a predictive model were proposed in this study, which could identify women who are at high risk of developing AUB-O. This finding highlights the importance of early screening and the lifelong management of ovulatory disorders for women.

Global, regional, and national time trends in incidence, prevalence, years lived with disability for uterine fibroids, 1990-2019: an age-period-cohort analysis for the global burden of disease 2019 study.

BACKGROUND: Uterine fibroids are the most common benign neoplasm of the uterus and a major source of morbidity for women. We report an overview of trends in uterine fibroids of incidence rate, prevalence rate, years lived with disability (YLDs) rate in 204 countries and territories over the past 30 years and associations with age, period, and birth cohort. METHODS: The incident case, incidence rate, age-standardized rate (ASR) for incidence, prevalent case, prevalence rate, ASR for prevalence, number of YLDs, YLD rate, and ASR for YLDs were derived from the Global Burden of Disease 2019 (GBD 2019) study. We utilized an age-period-cohort (APC) model to estimate overall annual percentage changes in the rate of incidence, prevalence, and YLDs (net drifts), annual percentage changes from 10 to 14 years to 65-69 years (local drifts), period and cohort relative risks (period/cohort effects) between 1990 and 2019. RESULTS: Globally, the incident cases, prevalent cases, and the number of YLDs of uterine fibroids increased from 1990 to 2019 with the growth of 67.07%, 78.82% and 77.34%, respectively. High Socio-demographic Index (SDI) and high-middle SDI quintiles with decreasing trends (net drift < 0.0%), and increasing trends (net drift > 0.0%) were observed in middle SDI, low-middle SDI, and low SDI quintiles in annual percentage change of incidence rate, prevalence rate and YLDs rate over the past 30 years. There were 186 countries and territories that showed an increasing trend in incidence rate, 183 showed an increasing trend in prevalence rate and 174 showed an increasing trend in YLDs rate. Moreover, the effects of age on uterine fibroids increased with age and peaked at 35-44 years and then declined with advancing age. Both the period and cohort effects on uterine fibroids showed increasing trend in middle SDI, low-middle SDI and low SDI quintiles in recent 15 years and birth cohort later than 1965. CONCLUSIONS: The global burden of uterine fibroids is becoming more serious in middle SDI, low-middle SDI and low SDI quintiles. Raising awareness of uterine fibroids, increasing medical investment and improving levels of medical care are necessary to reduce future burden.

YUCCA2 (YUC2)-Mediated 3-Indoleacetic Acid (IAA) Biosynthesis Regulates Chloroplast RNA Editing by Relieving the Auxin Response Factor 1 (ARF1)-Dependent Inhibition of Editing Factors in Arabidopsis thaliana.

Although recent research progress on the abundant C-to-U RNA editing events in plant chloroplasts and mitochondria has uncovered many recognition factors and their molecular mechanisms, the intrinsic regulation of RNA editing within plants remains largely unknown. This study aimed to establish a regulatory relationship in Arabidopsis between the plant hormone auxin and chloroplast RNA editing. We first analyzed auxin response elements (AuxREs) present within promoters of chloroplast editing factors reported to date. We found that each has more than one AuxRE, suggesting a potential regulatory role of auxin in their expression. Further investigation unveiled that the depletion of auxin synthesis gene YUC2 reduces the expression of several editing factors. However, in yuc2 mutants, only the expression of CRR4, DYW1, ISE2, and ECD1 editing factors and the editing efficiency of their corresponding editing sites, ndhD-2 and rps14-149, were simultaneously suppressed. In addition, exogenous IAA and the overexpression of YUC2 enhanced the expression of these editing factors and the editing efficiency at the ndhD-2 and rps14-149 sites. These results suggested a direct effect of auxin upon the editing of the ndhD-2 and rps14-149 sites through the modulation of the expression of the editing factors. We further demonstrated that ARF1, a downstream transcription factor in the auxin-signaling pathway, could directly bind to and inactivate the promoters of CRR4, DYW1, and ISE2 in a dual-luciferase reporter system, thereby inhibiting their expression. Moreover, the overexpression of ARF1 in Arabidopsis significantly reduced the expression of the three editing factors and the editing efficiency at the ndhD-2 and rps14-149 sites. These data suggest that YUC2-mediated auxin biosynthesis governs the RNA-editing process through the ARF1-dependent signal transduction pathway.

Enantiodivergent Hydrogenation of Exocyclic α,ß-Unsaturated Lactams Enabled by Switching the N-Chirality of Iridium Catalyst.

Central chirality is an important chiral element used in the design of chiral ligands and catalysts. Mostly, the attention of organic chemists is focused on developing of chiral ligands with stable stereogenic centers. However, the N-chirality in chiral ligand design has been rarely explored due to its flexibility. Here we demonstrate the design, synthesis, and application of a class of simple P,N-ligands with flexible N-chirality and their derived iridium complexes with fixed N-chiral stereocenters. Both fixed configurations of the N-stereocenter of the iridium complexes could be selectively formed from the same chiral ligand. This pair of diastereoisomeric iridium complexes showed good performance in the enantiodivergent asymmetric hydrogenation of exocyclic α,ß-unsaturated lactams. The N-H group plays an impressive role in catalytic activity. Computational studies emphasized the importance of N-chirality and N-H group.

Survival Nomogram for Young Breast Cancer Patients Based on the SEER Database and an External Validation Cohort.

BACKGROUND: Young breast cancer (YBC) patients are more prone to lymph node metastasis than other age groups. Our study aimed to investigate the predictive value of lymph node ratio (LNR) in YBC patients and create a nomogram to predict overall survival (OS), thus helping clinical diagnosis and treatment. METHODS: Patients diagnosed with YBC between January 2010 and December 2015 from the Surveillance, Epidemiology, and End Results (SEER) database were enrolled and randomly divided into a training set and an internal validation set with a ratio of 7:3. An independent cohort from our hospital was used for external validation. Univariate and least absolute shrinkage and selection operator (LASSO) regression were used to identify the significant factors associated with prognosis, which were used to create a nomogram for predicting 3- and 5-year OS. RESULTS: We selected seven survival predictors (tumor grade, T-stage, N-stage, LNR, ER status, PR status, HER2 status) for nomogram construction. The C-indexes in the training set, the internal validation set, and the external validation set were 0.775, 0.778 and 0.817, respectively. The nomogram model was well calibrated, and the time-dependent ROC curves verified the superiority of our model for clinical usefulness. In addition, the nomogram classification could more precisely differentiate risk subgroups and improve the discrimination of YBC prognosis. CONCLUSIONS: LNR is a strong predictor of OS in YBC patients. The novel nomogram based on LNR is a reliable tool to predict survival, which may assist clinicians in identifying high-risk patients and devising individual treatments.

An artificial pathway for trans-4-hydroxy-L-pipecolic acid production from L-lysine in Escherichia coli.

Trans-4-hydroxy-L-pipecolic acid (trans-4-HyPip) is a hydroxylated product of L-pipecolic acid, which is widely used in the pharmaceutical and chemical industries. Here, a trans-4-HyPip biosynthesis module was designed and constructed in Escherichia coli by overexpressing lysine α-oxidase, Δ1-piperideine-2-carboxylase reductase, glucose dehydrogenase, lysine permease, catalase and L-pipecolic acid trans-4-hydroxylase for expanding the lysine catabolism pathway. A total of 4.89 g/L of trans-4-HyPip was generated in shake flasks from 8 g/L of L-pipecolic acid. By this approach, 14.86 g/L of trans-4-HyPip was produced from lysine after 48 h in a 5 L bioreactor. As far as we know, this is the first multi-enzyme cascade catalytic system for the production of trans-4-HyPip using E. coli from L-lysine. Therefore, it can be considered as a potential candidate for the industrial production of trans-4-HyPip in microorganisms.

In Situ Visual Distribution of Gelsemine, Koumine, and Gelsenicine by MSI in Gelsemiumelegans at Different Growth Stages.

The distribution of pharmatically important alkaloids gelsemine, koumine, and gelsenicine in Gelsemium elegans tissues is a hot topic attracting research attention. Regretfully, the in planta visual distribution details of these alkaloids are far from clear although several researches reported the alkaloid quantification in G. elegans by LC-MS/MS. In this study, mass imaging spectrometry (MSI) was employed to visualize the in situ visualization of gelsemine, koumine, and gelsenicine in different organs and tissues of G. elegans at different growth stages, and the relative quantification of three alkaloids were performed according to the image brightness intensities captured by the desorption electrospray ionization MSI (DESI-MSI). The results indicated that these alkaloids were mainly accumulated in pith region and gradually decreased from pith to epidermis. Interestingly, three alkaloids were found to be present in higher abundance in the leaf vein. Along with the growth and development, the accumulation of these alkaloids was gradually increased in root and stem. Moreover, we employed LC-MS/MS to quantify three alkaloids and further validated the in situ distributions. The content of koumine reached 249.2 µg/g in mature roots, 272.0 µg/g in mature leaves, and 149.1 µg/g in mature stems, respectively, which is significantly higher than that of gelsemine and gelsenicine in the same organ. This study provided an accurately in situ visualization of gelsemine, koumine, and gelsenicine in G. elegans, and would be helpful for understanding their accumulation in plant and guiding application.

Panorama of intron dynamics and gene rearrangements in the phylum Basidiomycota as revealed by the complete mitochondrial genome of Turbinellus floccosus.

In the present study, the complete mitogenome of Turbinellus floccosus was sequenced, assembled, and compared with other basidiomycete mitogenomes. The mitogenome of T. floccosus consists of a circular DNA molecule, with a size of 62,846 bp. Gene arrangement analysis indicated that large-scale gene rearrangements occurred in the levels of family and genus of basidiomycete species, and the mitogenome of T. floccosus contained a unique gene order. A significant correlation between the number of introns and the mitochondrial genome size of Basidiomycota were detected (P < 0.01). A total of 896 introns were detected in the core protein-coding genes (PCGs) of 74 basidiomycete species, and the cox1 gene was the largest host gene of basidiomycete introns. Intron position class (Pcls) P383 in the cox1 gene was the most common intron in Basidiomycota, which distributed in 40 of 74 basidiomycete species. In addition, frequent intron loss/gain events were detected in basidiomycete species. More than 50% of bases around insertion sites (- 15 bp to 15 bp) of Pcls from different species were conservative, indicating site preferences of intron insertions in Basidiomycota. Further analysis showed that 76.09% of introns tended to insert downstream to a T base in Basidiomycota. Phylogenetic analysis for 74 basidiomycetes indicated mitochondrial genes are effective molecular markers for phylogeny of basidiomycetes. The study served as the first report on the mitogenome from the family Gomphaceae, which will help to understand the intron origin and evolution in Basidiomycota. KEY POINTS: • The mitogenome of Turbinellus floccosus had a unique gene arrangement. • Intron loss/gain events were detected in the 74 basidiomycete species. • Introns tend to insert downstream of a T base in basidiomycete mitogenomes.

Erythromycin Regulates Cigarette Smoke-Induced Proinflammatory Mediator Release Through Sirtuin 1-Nuclear Factor κB Axis in Macrophages and Mice Lungs.

BACKGROUND: Macrolides have anti-inflammatory and antioxidative stress function, but their pharmacological regulation remains unclear. Sirtuin 1 (SIRT1) is redox-sensitive protein belongs to class III histone/protein deacetylases, SIRT1 regulates the acetylation/expression of nuclear factor κB (NF-κB) and is involved in the airway inflammation of chronic obstructive pulmonary disease. OBJECTIVES: The present study was designed to examine the effects of erythromycin (EM) on the SIRT1-NF-κB axis and NF-κB-dependent proinflammatory cytokines. METHODS: Human macrophages were preincubated with EM and then treated with cigarette smoke extract (CSE). The mice were treated by injecting drugs to gastric with EM before cigarette smoke exposure. Reactive oxygen species (ROS) released by treated human macrophages were detected using flow cytometry. The expression of SIRT1 and NF-κB was analyzed by western blotting. SIRT1 and the RelA/p65 subunits of NF-κB interaction were detected by coimmunoprecipitation. We found that EM suppressed CSE-induced ROS released in human macrophages, which coincided with increases in SIRT1 protein expression in the macrophages and lungs of mice, resulting in suppressed -NF-κB acetylation and expression correlated with a reduction of inflammatory mediators. CONCLUSION: These findings suggest that EM increased SIRT1, leading to acetylation/expression of NF-κB, and thereby decreasing cigarette smoke-driven NF-κB-dependent proinflammatory cytokine.

TALEN-mediated precise genome modification by homologous recombination in zebrafish.

We report gene targeting via homologous recombination in zebrafish. We co-injected fertilized eggs with transcription activator-like effector nuclease mRNAs and a donor vector with long homologous arms targeting the tyrosine hydroxylase (th) locus, and we observed effective gene modification that was transmitted through the germ line. We also successfully targeted two additional genes. Homologous recombination in zebrafish with a dsDNA donor expands the utility of this model organism.

Chromosomal deletions and inversions mediated by TALENs and CRISPR/Cas in zebrafish.

Customized TALENs and Cas9/gRNAs have been used for targeted mutagenesis in zebrafish to induce indels into protein-coding genes. However, indels are usually not sufficient to disrupt the function of non-coding genes, gene clusters or regulatory sequences, whereas large genomic deletions or inversions are more desirable for this purpose. By injecting two pairs of TALEN mRNAs or two gRNAs together with Cas9 mRNA targeting distal DNA sites of the same chromosome, we obtained predictable genomic deletions or inversions with sizes ranging from several hundred bases to nearly 1 Mb. We have successfully achieved this type of modifications for 11 chromosomal loci by TALENs and 2 by Cas9/gRNAs with different combinations of gRNA pairs, including clusters of miRNA and protein-coding genes. Seven of eight TALEN-targeted lines transmitted the deletions and one transmitted the inversion through germ line. Our findings indicate that both TALENs and Cas9/gRNAs can be used as an efficient tool to engineer genomes to achieve large deletions or inversions, including fragments covering multiple genes and non-coding sequences. To facilitate the analyses and application of existing ZFN, TALEN and CRISPR/Cas data, we have updated our EENdb database to provide a chromosomal view of all reported engineered endonucleases targeting human and zebrafish genomes.

MicroRNAs delivered by extracellular vesicles: an emerging resistance mechanism for breast cancer.

Resistance to chemotherapy and endocrine therapy as well as targeted drugs is a major problem in treatment of breast cancer. Over the last decades, emerging studies have revealed that extracellular vesicles, which are chronically released by breast cancer cells and surrounding stromal cells, influence the action of most commonly used therapeutics. Such modulatory effects have been related to the transport of biologically active molecules including proteins and functional microRNAs. In this review, we highlight recent studies regarding extracellular vesicle-mediated microRNA delivery in formatting drug resistance. We also suggest the use of extracellular vesicles as a promising method in antiresistance treatment.

Research on the evolution of cross-platform online public opinion for public health emergencies considering stakeholders.

OBJECTIVE: To explore the different processes of the themes and emotional evolution of various stakeholders in the network public opinion of sudden public health emergencies at different stages of the public opinion evolution lifecycle. METHODS: This paper proposes a cross-platform analysis method for online public opinion during the public health emergencies based on stakeholders. Firstly, data from multiple platforms are collected and integrated. Secondly, stakeholders are categorized and the stages of public opinion evolution are determined based on stakeholder theory and lifecycle theory. Finally, the Latent Dirichlet Allocation (LDA)+Word2vec model and Convolutional Neural Network (CNN) model are used to analyze the themes and emotional evolution of stakeholders during different stages of public opinion evolution. RESULTS: There are differences in the evolution patterns of different types of stakeholders. The evolution process of stakeholders' focus points exhibits a two-stage transition from concentration to divergence. The focus points of stakeholders are closely associated with their respective social domains. The emotions of the public undergo a three-stage process of positive-negative-positive change. CONCLUSIONS: This study can provide a reference for the government to have a more comprehensive understanding of the development trend of public opinion and reduce the negative impact of public opinion.

Development and Validation of a Prognostic Nomogram for Breast Cancer Patients With Multi-Organ Metastases: An Analysis of the Surveillance, Epidemiology, and End Results Program Database.

BACKGROUND: Multi-organ metastases represent a substantial life-threatening risk for breast cancer (BC) patients. Nonetheless, the current dearth of assessment tools for patients with multi-organ metastatic BC adversely impacts their evaluation. METHODS: We conducted a retrospective analysis of BC patients with multi-organ metastases using data from the SEER database from 2010 to 2019. The patients were randomly allocated into a training cohort and a validation cohort in a 7:3 ratio. Univariate COX regression analysis, the LASSO, and multivariate Cox regression analyses were performed to identify independent prognostic factors in the training set. Based on these factors, a nomogram was constructed to estimate overall survival (OS) probability for BC patients with multi-organ metastases. The performance of the nomogram was evaluated using C-indexes, ROC curves, calibration curves, decision curve analysis (DCA) curves, and the risk classification system for validation. RESULTS: A total of 3626 BC patients with multi-organ metastases were included in the study, with 2538 patients in the training cohort and 1088 patients in the validation cohort. Age, grade, metastasis location, surgery, chemotherapy, and subtype were identified as significant independent prognostic factors for OS in BC patients with multi-organ metastases. A nomogram for predicting 1-year, 3-year, and 5-year OS was constructed. The evaluation metrics, including C-indexes, ROC curves, calibration curves, and DCA curves, demonstrated the excellent predictive performance of the nomogram. Additionally, the risk grouping system effectively stratified BC patients with multi-organ metastases into distinct prognostic categories. CONCLUSION: The developed nomogram showed high accuracy in predicting the survival probability of BC patients with multi-organ metastases, providing valuable information for patient counseling and treatment decision making.

Validation of CDC45 as a novel biomarker for diagnosis and prognosis of gastric cancer.

Background: Cell division cycle protein 45 (CDC45) has been demonstrated to play vital roles in the progression of various malignancies. However, the clinical significance of CDC45 in gastric cancer (GC) remains unreported. Method: In this study, we employed the TCGA database and the TCGA & GTEx dataset to compare the mRNA expression levels of CDC45 between gastric cancer tissues and adjacent or normal tissues (p < 0.05 was considered statistically significant), which was further validated in multiple datasets including GSE13911, GSE29272, GSE118916, GSE66229, as well as RT-qPCR. Furthermore, we harnessed the Human Protein Atlas (HPA) to evaluate the protein expression of CDC45, which was subsequently verified through immunohistochemistry (IHC). To ascertain the diagnostic utility of CDC45, receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC) were calculated in TCGA database, and further validated it in TCGA & GTEx and GSE66229 datasets. The Kaplan-Meier method was used to reveal the prognostic importance of CDC45 in The Cancer Genome Atlas (TCGA) database and authenticated through the GSE66229, GSE84433, and GSE84437 datasets. Through cBioPortal, we identified co-expressed genes of CDC45, and pursued enrichment analysis. Additionally, we availed gene set enrichment analysis (GSEA) to annotate the biological functions of CDC45. Results: Differential expression analysis revealed that CDC45 was significantly upregulated at both the mRNA and protein levels in GC (all p < 0.05). Remarkably, CDC45 emerged as a promising prognostic indicator and a novel diagnostic biomarker for GC. In a comprehensive the drug susceptibility analysis, we found that patients with high expression of CDC45 had high sensitivity to various chemotherapeutic agents, among which 5-fluorouracil, docetaxel, cisplatin, and elesclomol were most evident. Furthermore, our findings suggested a plausible association between CDC45 and immune cell infiltration. Enrichment analysis revealed that CDC45 and its associated genes may play crucial roles in muscle biofunction, whereas GSEA demonstrated significant enrichment of gene sets pertaining to G protein-coupled receptor ligand binding and G alpha (i) signaling events. Conclusion: Our study elucidates that upregulation of CDC45 is intricately associated with immune cell infiltration and holds promising potential as a favorable prognostic marker and a novel diagnostic biomarker for GC.

Vaginal microbiota, menopause, and the use of menopausal hormone therapy: a cross-sectional, pilot study in Chinese women.

OBJECTIVE: To compare the vaginal microbiota of premenopausal and postmenopausal women and postmenopausal women undergoing menopausal hormone therapy (MHT) and examine the association between vaginal microbiota and genitourinary syndrome of menopause (GSM). METHODS: This cross-sectional study classified 94 women aged 40 to 60 years into three groups: premenopausal (Pre, n = 32), postmenopausal (Post, n = 30), and postmenopausal women who received MHT orally (Post + MHT, n = 32). Neither the Pre nor the Post group received hormone therapy within the past 6 months. Postmenopausal women who received vaginal MHT were not included. Vaginal swabs were obtained, and microbial composition was characterized by 16S rRNA gene sequencing targeting the V3 to V4 region. Clinical data were collected and serum sex hormones were measured. The most bothersome symptom approach and vaginal health index were used to evaluate GSM. Mann-Whitney U or Kruskal-Wallis ANOVA followed by multiple comparison tests were performed for comparison between or across groups. The correlations between GSM symptom scores and vaginal microbiota were determined using Spearman's correlation analysis. RESULTS: The vaginal community of postmenopausal women was characterized by a decreased abundance of Lactobacillus (Post 18% vs Pre 69%); an increased abundance of several anaerobic bacteria, including Prevotella, Escherichia-Shigella, and Bifidobacterium; and a higher microbial diversity (P < 0.001 for Shannon and Simpson indexes) than those of premenopausal women. The vaginal community of postmenopausal women who received MHT had an increased abundance of Lactobacillus (54%) and lower microbial diversity (P < 0.001 for Shannon and Simpson indexes) than the postmenopausal women. The vaginal microbial community composition of the Pre group shared more similarity with that of the Post + MHT group (Adonis P = 0.051) than with that of the Post group (Adonis P < 0.001). A decreased abundance of Lactobacillus and high diversity in the vaginal community were found in women with moderate to severe GSM symptoms. CONCLUSIONS: Among Chinese postmenopausal women, those receiving MHT had higher Lactobacillus abundance but lower abundance of diverse anaerobes and diversity of the vaginal microbial community compared to non-MHT women. MHT in postmenopausal women may potentially contribute to reestablishing vaginal microbiota homeostasis. Findings in this pilot study, however, need to be examined in larger, prospective studies.