The CRL3KCTD10 ubiquitin ligase-USP18 axis coordinately regulates cystine uptake and ferroptosis by modulating SLC7A11.

SLC7A11 is a cystine transporter and ferroptosis inhibitor. How the stability of SLC7A11 is coordinately regulated in response to environmental cystine by which E3 ligase and deubiquitylase (DUB) remains elusive. Here, we report that neddylation inhibitor MLN4924 increases cystine uptake by causing SLC7A11 accumulation, via inactivating Cullin-RING ligase-3 (CRL-3). We identified KCTD10 as the substrate-recognizing subunit of CRL-3 for SLC7A11 ubiquitylation, and USP18 as SLC7A11 deubiquitylase. Upon cystine deprivation, the protein levels of KCTD10 or USP18 are decreased or increased, respectively, contributing to SLC7A11 accumulation. By destabilizing or stabilizing SLC7A11, KCTD10, or USP18 inversely regulates the cystine uptake and ferroptosis. Biologically, MLN4924 combination with SLC7A11 inhibitor Imidazole Ketone Erastin (IKE) enhanced suppression of tumor growth. In human breast tumor tissues, SLC7A11 levels were negatively or positively correlated with KCTD10 or USP18, respectively. Collectively, our study defines how SLC7A11 and ferroptosis is coordinately regulated by the CRL3KCTD10/E3-USP18/DUB axis, and provides a sound rationale of drug combination to enhance anticancer efficacy.

A Multistage Hemiplegic Lower-Limb Rehabilitation Robot: Design and Gait Trajectory Planning.

Most lower limb rehabilitation robots are limited to specific training postures to adapt to stroke patients in multiple stages of recovery. In addition, there is a lack of attention to the switching functions of the training side, including left, right, and bilateral, which enables patients with hemiplegia to rehabilitate with a single device. This article presents an exoskeleton robot named the multistage hemiplegic lower-limb rehabilitation robot, which has been designed to do rehabilitation in multiple training postures and training sides. The mechanism consisting of the thigh, calf, and foot is introduced. Additionally, the design of the multi-mode limit of the hip, knee, and ankle joints supports delivering therapy in any posture and training sides to aid patients with hemiplegia in all stages of recovery. The gait trajectory is planned by extracting the gait motion trajectory model collected by the motion capture device. In addition, a control system for the training module based on adaptive iterative learning has been simulated, and its high-precision tracking performance has been verified. The gait trajectory experiment is carried out, and the results verify that the trajectory tracking performance of the robot has good performance.

Glucose metabolic reprogramming and its therapeutic potential in obesity-associated endometrial cancer.

Endometrial cancer (EC) is a common gynecological cancer that endangers women health. Although substantial progresses of EC management have been achieved in recent years, the incidence of EC still remains high. Obesity has been a common phenomenon worldwide that increases the risk of EC. However, the mechanism associating obesity and EC has not been fully understood. Metabolic reprogramming as a remarkable characteristic of EC is currently emerging. As the primary factor of metabolic syndrome, obesity promotes insulin resistance, hyperinsulinemia and hyperglycaemia. This metabolic disorder remodels systemic status, which increases EC risk and is related with poor prognosis. Glucose metabolism in EC cells is complex and mediated by glycolysis and mitochondria to ensure energy requirement. Factors that affect glucose metabolism may have an impact on EC initiation and progression. In this study, we review the glucose metabolic reprogramming of EC not only systemic metabolism but also inherent tumor cell metabolism. In particular, the role of glucose metabolic regulation in malignant properties of EC will be focused. Understanding of metabolic profile and glucose metabolism-associated regulation mechanism in EC may provide novel perspective for treatment.

The relationship between body dissatisfaction, lifestyle, and nutritional status among university students in Southern China.

BACKGROUND: In recent years, obesity in early adulthood has become an urgent global public health concern. Body dissatisfaction may have adverse effects on lifestyle habits, leading to obesity. However, research on nutritional status and body dissatisfaction among Chinese young adults is still insufficient. Therefore, this study aimed to analyze the relationship between body dissatisfaction, dietary habits, physical activity, and nutritional status among university students. In addition, we explored the feasibility of improving university students' nutritional status by improving the levels of body dissatisfaction. METHODS: This study was conducted in Ganzhou City, Jiangxi Province, China, at a randomly selected university. All 1900 undergraduate students volunteered to participate and signed the consent form. Students were required to completed anthropometric measurements and three questionnaires, which included the Physical Activity Rating Scale-3 (PARS-3), Chinese version of the Dutch Dietary Behavior Questionnaire (C-DEBQ), and Body Dissatisfaction. Of these, 1714 students (age: 18-24 years; men: 933, women: 781) with complete and valid data were included. RESULTS: Higher obesity levels were observed in men compared to women (p<0.01). Meanwhile, body dissatisfaction was higher in women compared to men (p<0.01). Overeating and insufficient physical activity were more problematic in women compared to in men (p<0.01). Multiple regression analyses were conducted separately, with BMI and body dissatisfaction as the dependent variables. Body dissatisfaction (ß=0.72, p<0.01), muscle mass (ß=0.33, p<0.01), emotional eating score (ß=0.05, p<0.01), sex (ß=-0.05, p<0.05) and physical activity (ß=-0.04, p<0.05) score were significant predictors of obesity. Furthermore, Muscle mass (ß=0.61, p<0.01), sex (ß=0.54, p<0.01), restrained eating score (ß=0.25, p<0.01), physical activity score (ß=-0.20, p<0.01) and emotional eating score (ß=0.08, p<0.01) were significant predictors of body dissatisfaction. CONCLUSION: The data presented in this study highlight the impact of university students' body dissatisfaction in China on physical activity deficiency and overeating, discovering that reducing body dissatisfaction has great potential for preventing obesity.

Tumor-suppressive circRHOBTB3 is excreted out of cells via exosome to sustain colorectal cancer cell fitness.

BACKGROUND & AIMS: To clarify the biological roles, circularization process and secretion pathway of circRHOBTB3 in colorectal cancer (CRC) progression. METHODS: We performed a comprehensive analysis of circRNA levels in serum exosomes from multiple types of cancer patients in public databases and verified the higher level of circRHOBTB3 in CRC sera versus healthy donors by RT-qPCR. Then, the function of circRHOBTB3 in CRC was investigated in vitro and in vivo. RNA-seq and RNA pull-down assays together with mass spectrometry identified the downstream signals and the binding proteins of circRHOBTB3. Finally, Antisense oligonucleotides (ASOs) were designed to target circularization and secretion elements of circRHOBTB3 for CRC therapy. RESULTS: circRHOBTB3 levels were increased in the sera but was downregulated in tissue samples in CRC, and the downregulation was associated with poor prognosis. Furthermore, circRHOBTB3 acts a tumor-suppressive circRNA by repressing metabolic pathways, intracellular ROS production in CRC. Several key elements were discovered to regulate circRHOBTB3 circularization and exosomal secretion. Moreover, SNF8 was identified that sorts circRHOBTB3 into exosomes. Interestingly, we found that CRC cells could actively secrete more circRHOBTB3 than normal cells. According to the sequence of regulatory elements for circularization and exosomal secretion, we designed and synthesized ASOs, which increased circRHOBTB3 expression and blocked circRHOBTB3 exosomal secretion. More importantly, ASOs could inhibit CRC growth and metastasis in vitro and in vivo. CONCLUSIONS: circRHOBTB3 plays a tumor-suppressive role in CRC and has to be excreted out of cells to sustain cancer cell fitness. ASOs targeting regulatory elements for circularization and exosomal secretion will become a novel antitumor strategy.

Sexual debut among college students in China: effects of family context.

This study examines family context and sexual debut among young people in China. Using data from the 2018 Panel Study of Chinese University Students (PSCUS), it explores how the family is correlated with sexual debut among young people in China aged 18-24 years. The Kaplan-Meier method was adopted to detect a survival function for different family factors and related demographic variables. Cox proportional hazard regression analysis was adopted to calculate hazard ratios for the timing of sexual debut. The average age of sexual debut among the college students was 18.39 years. The Kaplan-Meier analysis showed that sexual intercourse initiation was earlier for female students who had no siblings, and those who had a mother with senior high school (including technical school) education or higher family income, but this correlation was insignificant among male students. The multivariate hazard regression analysis revealed that living in a family with a higher level of fathers' education, having a lower level of family income and having siblings had positive correlations with later sexual debut among the college students. Moreover, family factors showed gender differences in their associations with the timing of sexual debut, typically parent's education level, family income and left-behind experience. This study provides a comprehensive perspective on the role of family influences in timing of sexual debut among youth in China.

Distinct roles of programmed death ligand 1 alternative splicing isoforms in colorectal cancer.

Although anti-programmed death-1 (PD-1)/programmed death ligand 1 (PD-L1) immunotherapy has achieved great success in some cancers, most colorectal cancer (CRC) patients remain unresponsive. Therefore, further clarification of the underlying mechanisms is needed to improve the therapy. In this study, we explored the distinct functions of different PD-L1 alternative splicing isoforms in CRC. We investigated the biological functions in PD-L1 knocked down/knockout cells, which were verified through overexpression of PD-L1 isoforms a, b, and c. The roles of PD-L1 isoforms in immune surveillance resistance was also analyzed. Meanwhile, we performed RNA-seq to screen the downstream molecules regulated by PD-L1 isoforms. Finally, we detected PD-L1 and PD-L1 isoforms levels in a cohort of serum samples, two cohorts of CRC tissue samples, and analyzed the correlation of PD-L1 isoforms with PD-1 blockade therapy response in two clinical CRC cases. The results indicated that PD-L1 knockout inhibited proliferation, migration, and invasion, and isoform b exerted a more significant inhibitory effect on T cells than the other two isoforms. Moreover, isoform c could promote CRC progression through regulating epithelial-mesenchymal transition. Clinical data showed that CRC patients with positive PD-L1 expression were associated with poorer overall survival. High serum PD-L1 level was associated with poor prognosis. The level of isoform b or c was negatively associated with prognosis, and a higher level of isoform b was associated with a good response to anti-PD-1 therapy. In conclusion, isoform b should be considered as a biomarker for clinical responsiveness to anti-PD-1/PD-L1 immunotherapy; isoform c had a prometastatic role and is a new potential target for CRC therapy.

The long non-coding RNA CYTOR drives colorectal cancer progression by interacting with NCL and Sam68.

BACKGROUND: Long non-coding RNAs (lncRNAs) function as key molecules in cancer progression. The lncRNA CYTOR plays oncogenic roles in multiple types of cancer, yet the detailed molecular mechanisms of those roles remain unknown. The aim of this study was to investigate the clinical significance, biological function and interacting partners of CYTOR in colorectal cancer (CRC). METHODS: A systematic and comprehensive analysis of CYTOR expression was performed in 138 CRC samples and in the TCGA and GEO databases. Biological function was investigated through knockdown and overexpression of CYTOR in vitro and in vivo. In addition, its protein binding partner was identified and validated using ChIRP-MS and RNA immunoprecipitation assays. Their key interaction sites on CYTOR were verified by CRISPR/Cas9 and a series of mutant constructs. Furthermore, the downstream targets of CYTOR were confirmed via immunoblotting and luciferase reporter assays. RESULTS: CYTOR was significantly up-regulated in CRC samples and associated with poor prognosis, promoting proliferation and metastasis in vitro and in vivo. NCL and Sam68 could recognize their specific motifs and directly bind to EXON1 of CYTOR. Moreover, EXON1 was the key functional site mediating the interaction of CYTOR with NCL and Sam68. NCL and Sam68 functioned as oncogenes to promote CRC progression. Furthermore, we confirmed that the heterotrimeric complex of CYTOR, NCL and Sam68 activated the NF-κB pathway and EMT to contribute to CRC progression. CONCLUSION: CYTOR plays important roles in CRC progression by interacting with NCL and Sam68 and may serve as a prognostic biomarker and/or an effective target for CRC therapies.

Metabolism mechanisms of biogenic methane production by synergistic biodegradation of lignite and guar gum.

Coalbed methane (CBM) presents a promising energy source for addressing global energy shortages. Nonetheless, challenges such as low gas production from individual wells and difficulties in breaking gels at low temperatures during extraction hinder its efficient utilization. Addressing this, we explored native microorganisms within coal seams to degrade guar gum, thereby enhancing CBM production. However, the underlying mechanisms of biogenic methane production by synergistic biodegradation of lignite and guar gum remain unclear. Research results showed that the combined effect of lignite and guar gum enhanced the production, yield rate and concentration of biomethane. When the added guar gum content was 0.8 % (w/w), methane production of lignite and guar gum reached its maximum at 561.9 mL, which was 11.8 times that of single lignite (47.3 mL). Additionally, guar gum addition provided aromatic and tryptophan proteins and promoted the effective utilization of CC/CH and OCO groups on the coal surface. Moreover, the cooperation of lignite and guar gum accelerated the transformation of volatile fatty acids into methane and mitigated volatile fatty acid inhibition. Dominant bacteria such as Sphaerochaeta, Macellibacteroides and Petrimonas improved the efficiency of hydrolysis and acidification. Electroactive microorganisms such as Sphaerochaeta and Methanobacterium have been selectively enriched, enabling the establishment of direct interspecies electron transfer pathways. This study offers valuable insights for increasing the production of biogenic CBM and advancing the engineering application of microbial degradation of guar gum fracturing fluid. Future research will focus on exploring the methanogenic capabilities of lignite and guar gum in in-situ environments, as well as elucidating the specific metabolic pathways involved in their co-degradation.

Microbial community succession and volatile compounds changes during spontaneous fermentation of Cabernet Sauvignon (Vitis vinifera L.) under rain-shelter cultivation.

Microbiota succession in spontaneous fermentation of Cabernet Sauvignon cultivated under the rain-shelter was characterized, with open-field cultivation as the control. For both cultivation modes, Saccharomyces, Starmerella, and Mycosphearella were the principal fungi, and Tatumella, Gluconobacter, and Acinetobacter were the prevailing bacteria. Rain-shelter reduced the abundance of Hanseniaspora, Candida, Starmerella, Gluconobacter, and Lactococcus. During fermentation, fungal microbiota diversity in samples from the rain-shelter cultivation decreased more drastically than the control (p < 0.05). In terms of the correlation between microbiota and volatile compounds production, the abundance of Hanseniaspora uvarum, Candida apicola, Starmerella bacillaris, Gluconobacter oxydans, and Lactococcus lactis were positively correlated with the production of esters and higher alcohols. Instead of bacterial microbiota, fungal community succession exhibited a positive correlation with the final wine volatiles under the rain-shelter cultivation. These findings demonstrated rain-shelter cultivation influences the succession pattern of microbial communities and in turn impacts the wine aromas and flavors.

Leveraging 16S rRNA data to uncover vaginal microbial signatures in women with cervical cancer.

Microbiota-relevant signatures have been investigated for human papillomavirus-related cervical cancer (CC), but lack consistency because of study- and methodology-derived heterogeneities. Here, four publicly available 16S rRNA datasets including 171 vaginal samples (51 CC versus 120 healthy controls) were analyzed to characterize reproducible CC-associated microbial signatures. We employed a recently published clustering approach called VAginaL community state typE Nearest CentroId clAssifier to assign the metadata to 13 community state types (CSTs) in our study. Nine subCSTs were identified. A random forest model (RFM) classifier was constructed to identify 33 optimal genus-based and 94 species-based signatures. Confounder analysis revealed confounding effects on both study- and hypervariable region-associated aspects. After adjusting for confounders, multivariate analysis identified 14 significantly changed taxa in CC versus the controls (P < 0.05). Furthermore, predicted functional analysis revealed significantly upregulated pathways relevant to the altered vaginal microbiota in CC. Cofactor, carrier, and vitamin biosynthesis were significantly enriched in CC, followed by fatty acid and lipid biosynthesis, and fermentation of short-chain fatty acids. Genus-based contributors to the differential functional abundances were also displayed. Overall, this integrative study identified reproducible and generalizable signatures in CC, suggesting the causal role of specific taxa in CC pathogenesis.

Methyl radical chemistry in non-oxidative methane activation over metal single sites.

Molybdenum supported on zeolites has been extensively studied as a catalyst for methane dehydroaromatization. Despite significant progress, the actual intermediates and particularly the first C-C bond formation have not yet been elucidated. Herein we report evolution of methyl radicals during non-oxidative methane activation over molybdenum single sites, which leads selectively to value-added chemicals. Operando X-ray absorption spectroscopy and online synchrotron vacuum ultraviolet photoionization mass spectroscopy in combination with electron microscopy and density functional theory calculations reveal the essential role of molybdenum single sites in the generation of methyl radicals and that the formation rate of methyl radicals is linearly correlated with the number of molybdenum single sites. Methyl radicals transform to ethane in the gas phase, which readily dehydrogenates to ethylene in the absence of zeolites. This is essentially similar to the reaction pathway over the previously reported SiO2 lattice-confined single site iron catalyst. However, the availability of a zeolite, either in a physical mixture or as a support, directs the subsequent reaction pathway towards aromatization within the zeolite confined pores, resulting in benzene as the dominant hydrocarbon product. The findings reveal that methyl radical chemistry could be a general feature for metal single site catalysis regardless of the support (either zeolites MCM-22 and ZSM-5 or SiO2) whereas the reaction over aggregated molybdenum carbide nanoparticles likely facilitates carbon deposition through surface C-C coupling. These findings allow furthering the fundamental insights into non-oxidative methane conversion to value-added chemicals.

Tolerance for Housing Unaffordability among Highly Skilled Young Migrants: Evidence from the Zhejiang Province of China.

Many studies have concluded that, since housing pressure affects the mobility of highly skilled young migrants (HSYMs) in Chinese cities and regions, it is necessary to apply corresponding housing policies to adjust housing unaffordability for HYSM. This study uses data from a survey conducted in China's Zhejiang Province, where specific policies have been implemented to attract talent. We found that housing crowds out HSYM from a city, but that the HSYM who have a master's degree or above, or who work in government organizations or state-owned enterprises, are more tolerant of housing unaffordability. Those who are unmarried or those staying in the city for a long period are less tolerant of housing unaffordability. Meanwhile, different factors have heterogeneous impacts on the HSYMs' tolerance for housing unaffordability across cities of different levels. Therefore, housing policies should highlight urban differences and intra-group differences, and more housing land should be provided to attract talent.

Evaluation of rain-shelter cultivation mode effects on microbial diversity during Cabernet Sauvignon (Vitis vinifera L.) maturation in Jingyang, Shaanxi, China.

Rainfall particularly under continental climates with monsoonal tendency impacts the vineyard microbial niches during grapevine growth. With microbial community shifts, vine traits (grape flavor and yield) cultivated/protected under rain-shelter may ultimately be altered. Such cultivation may influence microflora dynamics via meteorological parameter variations, however this is unclear yet. Here, we used Cabernet Sauvignon, a prevalent red cultivar among wine growing regions, to evaluate the effects of the rain-shelter cultivation on the microorganism diversity. We found that average air temperature under rain-shelter conditions was 2-3 °C higher than the non-covered group, while air humidity the maximum reduction was 5.79% (p < 0.05). After grape setting stage, similar trends were observed on soil temperature (increased) and humidity (lowered) under the treatments (p < 0.05). UV and precipitation of rain-shelter treatment were less by a total of 72% and 96%, respectively (p < 0.05). The rain-shelter management presented lower fungal and bacterial OTUs. The fungal alpha diversity on leaves and branches under rain-shelter was lower (p < 0.05) than the control as the grape ripeness, with Ascomycota, Mycosphaerella and Cladosporium as the principal fungi. Our results revealed that the fungal microbiota patterns were differentiated by the cultivations from setting stage to the entire véraison and then tended to be similar at harvesting. Only branch fungal patterns were observed asymmetrically at all stages. Meanwhile, bacterial diversity and distribution varied on colonization locations where Proteobacteria and Actinobacteria were the primary bacteria phyla. Bacterial community structures overlapped at harvest, while the differences were observed between two cultivations at other stages, excluding grape berry. The rain-shelter cultivation reduced the abundance of Alternaria and Colletotrichum that may adversely affect grapevine health. Multivariate statistical analysis suggested that the effect of vineyard microclimate on microbiota distribution and succession were influenced by cultivation modes and grapevine developmental stages. This research provides evidence to address the dynamics of microbial ecology from vineyard to grape under rain-shelter cultivation, and its benefits as a sustainable vineyard management.

Long Noncoding RNA MIR4435-2HG Suppresses Colorectal Cancer Initiation and Progression By Reprogramming Neutrophils.

MIR4435-2HG, also known as LINC00978, has previously been described as an oncogenic long noncoding RNA (lncRNA). However, we show here that Mir4435-2hg depletion promoted colorectal tumorigenesis and progression in in vivo models of colitis-associated colorectal cancer, spontaneous intestinal adenomatous polyposis, and subcutaneous tumors. Alteration of MIR4435-2HG in colorectal cancer cells did not change the potential for cell proliferation, migration, or invasion in vitro. RNAscope assays showed that most MIR4435-2HG was located in the tumor stroma, which caused high expression of MIR4435-2HG in colorectal cancer tumor tissue. Transcriptome analysis of colorectal cancer tissues from wild-type and Mir4435-2hg-deficient mice revealed Mir4435-2hg as a tumor suppressor gene that regulated the immune microenvironment. Loss of Mir4435-2hg led to a decline in neutrophils and elevation of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC). In tissue-specific Mir4435-2hg knockout mice, we confirmed that Mir4435-2hg depletion in neutrophils, but not in intestinal epithelial cells, promoted colorectal cancer progression. Mechanistically, Mir4435-2hg depletion enhanced the immunosuppressive ability of PMN-MDSCs by disturbing their fatty acid metabolism. These findings suggest that MIR4435-2HG is a tumor-suppressing lncRNA whose deficiency could increase tumor-infiltrating PMN-MDSCs and enhance the immunosuppressive potential of PMN-MDSCs to promote colorectal cancer development. This provides a theoretical basis for further illustrating the pathogenesis of colorectal cancer and a potential antitumor immunotherapy target.

Disturbances of Vaginal Microbiome Composition in Human Papillomavirus Infection and Cervical Carcinogenesis: A Qualitative Systematic Review.

Background: Emerging evidence has demonstrated a close association between perturbations in vaginal microbiota composition in women and human papillomavirus (HPV) infection, cervical lesions, and cervical cancer (Ca); however, these findings are highly heterogeneous and inconclusive. Aim: To perform a comprehensive systematic review of the global disturbance in the vaginal microbiota, specifically in women with HPV-associated cervical diseases, and to further conduct within- and across-disease comparisons. Method: Twenty-two records were identified in a systematic literature search of PubMed, Web of Science, and Embase up to February 28, 2022. We extracted microbial changes at the community (alpha and beta diversity) and taxonomic (relative abundance) levels. Within- and across-disease findings on the relative abundance of taxonomic assignments were qualitatively synthesized. Results: Generally, significantly higher alpha diversity was observed for HPV infection, cervical lesions, and/or cancer patients than in controls, and significant differences within beta diversity were observed for the overall microbial composition across samples. In within-disease comparisons, the genera Gardnerella, Megasphaera, Prevotella, Peptostreptococcus, and Streptococcus showed the greatest abundances with HPV infection; Sneathia and Atopobium showed inconsistent abundance with HPV infection, and Staphylococcus was observed in Ca. Across diseases, we find increased levels of Streptococcus and varying levels of Gardnerella were shared across HPV infections, high-grade squamous intraepithelial lesions, and Ca, whereas Lactobacillus iners varied depending on the HPV-related disease subtype. Conclusions: This systematic review reports that vaginal microbiome disturbances are correlated to the depletion of Lactobacillus, enrichment of anaerobes, and increased abundance of aerobic bacteria in HPV infection and related cervical diseases. Moreover, L. iners may exert either protective or pathogenic effects on different HPV-related diseases.

Reference Genes for Expression Analyses by qRT-PCR in Phthorimaea operculella (Lepidoptera: Gelechiidae).

Due to a lack of effective internal references, studies on functional genes in Phthorimaea operculella, a serious Lepidopteran pest attacking potatoes worldwide, have been greatly limited. To select suitable endogenous controls, ten housekeeping genes of actin (ACT), α-tubulin (α-TUB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor 1α (EF1α), 18S and 28S ribosomal RNA (18S, 28S), ribosomal protein genes RPL4, RPL13 and RPL27 and superoxide dismutase (SOD) were tested. Their expression levels were determined under three different experimental conditions (developmental stages, tissues/organs and temperatures) using qRT-PCR technology. The stability was evaluated with five methods (Ct value, geNorm, NormFinder, BestKeeper and RefFinder). The results clarified that RPL13, EF1α and RPL27 are ranked as the best reference gene combination for measuring gene expression levels among different developing stages and under various temperatures; EF1α and RPL13 are recommended to normalize the gene expression levels among diverse tissues. EF1α and RPL13 are the best reference genes in all the experimental conditions. To validate the utility of the selected reference pair, EF1α and RPL13, we estimated the tissue-biased expression level of chitin synthase A gene (PoChSA). As expected, PoChSA was abundantly expressed in ectodermally derived epidermal cells, and lowly transcribed in the midgut. These findings will lay the foundation for future research on the molecular physiology and biochemistry of P. operculella.

Patterns of lymph node metastases and their implications in individualized radiotherapeutic clinical target volume delineation of regional lymph nodes in patients with endometrial cancer.

Purpose: To study the patterns of lymph node metastasis (LNM) of endometrial cancer (EC) and to clarify the individualized clinical target volume delineations of regional lymph nodes (CTVn). Methods: Data from a total of 556 patients with EC who had undergone total hysterectomy associated with bilateral salpingo-oophorectomy (TH/BSO) and systematic lymphadenectomy were retrospectively examined. The clinicopathological factors related to LNM were analyzed using logistic regression analysis. Results: LNM was found in 76 of 556 patients, resulting in a metastasis rate of 13.67%. The rates of LNM in patients with fundus and cornua lesions were 7.56% for para-aortic nodes and 14.41% for pelvic lymph nodes. The rates of LNM in patients with sidewall lesions were 3.15% for para-aortic nodes and 10.22% for pelvic lymph nodes. The rates of LNM in patients with lower uterine segment and cervix lesions were 12.50% for para-aortic nodes and 26.67% for pelvic lymph nodes. Deep myometrial invasion, histological type, histological differentiation, and lymphovascular space invasion (LVSI) emerged as statistically significant risk factors for pelvic LNM of EC (P = 0.008, 0.015, < 0.001, 0.005, respectively). Grade 3 differentiation had a strong influence on LNM to the para-aortic nodes (P = 0.015). Conclusions: Myometrial invasion, histological type, histological differentiation, and LVSI were related to pelvic LNM and grade 3 was associated with para-aortic LNM. These factors should be considered comprehensively to design the CTVn for intensity-modulated radiation therapy (IMRT) of EC. For patients with lower uterine segment/cervix and fundus/cornua lesions, delineating the irradiation field of the para-aortic nodal region may confer a benefit.

ENO2 Promotes Colorectal Cancer Metastasis by Interacting with the LncRNA CYTOR and Activating YAP1-Induced EMT.

The glycolytic enzyme enolase 2 (ENO2) is dysregulated in many types of cancer. However, the roles and detailed molecular mechanism of ENO2 in colorectal cancer (CRC) metastasis remain unclear. Here, we performed a comprehensive analysis of ENO2 expression in 184 local CRC samples and samples from the TCGA and GEO databases and found that ENO2 upregulation in CRC samples was negatively associated with prognosis. By knocking down and overexpressing ENO2, we found that ENO2 promoted CRC cell migration and invasion, which is dependent on its interaction with the long noncoding RNA (lncRNA) CYTOR, but did not depend on glycolysis regulation. Furthermore, CYTOR mediated ENO2 binding to large tumor suppressor 1 (LATS1) and competitively inhibited the phosphorylation of Yes-associated protein 1 (YAP1), which ultimately triggered epithelial-mesenchymal transition (EMT). Collectively, these findings highlight the molecular mechanism of the ENO2-CYTOR interaction, and ENO2 could be considered a potential therapeutic target for CRC.

LncRNA CASC15, MiR-23b Cluster and SMAD3 form a Novel Positive Feedback Loop to promote Epithelial-Mesenchymal Transition and Metastasis in Ovarian Cancer.

Cancer Susceptibility Candidate 15 (CASC15), which is a newly identified long noncoding RNA crucial for epigenetic regulation in human tumors, was found to be associated with poor prognosis of the patients with ovarian cancer by utilizing The Cancer Genome Atlas and Gene Expression Omnibus database. Therefore, the purpose of this paper was to explore the functional role and latent molecular mechanism of CASC15 in the progression of ovarian cancer. In vitro and in vivo experiments validated CASC15 as an oncogenic lncRNA in ovarian cancer, which could enhance metastasis through TGF-ß-induced epithelial-mesenchymal transition progress. MiR-23b-3p and miR-24-3p, which are members of the miR-23b cluster, were identified to directly target CASC15 through luciferase assays. Further mechanistic investigations indicated that CASC15-mediated miR-23b-3p/miR-24-3p sequestration cooperatively upregulated SMAD3 expression, which, in turn, would permit increased CASC15 mRNA level as a transcription activation factor. This study first described a miR-23b-3p/miR-24-3p-mediated positive feedback loop between CASC15 and SMAD3, which may reflect the underlying molecular mechanism of CASC15's oncogenic function in ovarian cancer.