ABSTRACT Purpose: The epithelial–mesenchymal transition of human lens epithelial cells plays a role in posterior capsule opacification, a fibrotic process that leads to a common type of cataract. Hyaluronic acid has been implicated in this fibrosis. Studies have investigated the role of transforming growth factor (TGF)-β2 in epithelial–mesenchymal transition. However, the role of TGF-β2 in hyaluronic acid-mediated fibrosis of lens epithelial cell remains unknown. We here examined the role of TGF-β2 in the hyaluronic acid-mediated epithelial–mesenchymal transition of lens epithelial cells. Methods: Cultured human lens epithelial cells (HLEB3) were infected with CD44-siRNA by using the Lipofectamine 3000 transfection reagent. The CCK-8 kit was used to measure cell viability, and the scratch assay was used to determine cell migration. Cell oxidative stress was analyzed in a dichloro-dihydro-fluorescein diacetate assay and by using a flow cytometer. The TGF-β2 level in HLEB3 cells was examined through immunohistochemical staining. The TGF-β2 protein level was determined through western blotting. mRNA expression levels were determined through quantitative real-time polymerase chain reaction. Results: Treatment with hyaluronic acid (1.0 μM, 24 h) increased the epithelial–mesenchymal transition of HLEB3 cells. The increase in TGF-β2 levels corresponded to an increase in CD44 levels in the culture medium. However, blocking the CD44 function significantly reduced the TGF-β2-mediated epithelial–mesenchymal transition response of HLEB3 cells. Conclusions: Our study showed that both CD44 and TGF-β2 are critical contributors to the hyaluronic acid-mediated epithelial-mesenchymal transition of lens epithelial cells, and that TGF-β2 in epithelial-mesenchymal transition is regulated by CD44. These results suggest that CD44 could be used as a target for preventing hyaluronic acid-induced posterior capsule opacification. Our findings suggest that CD44/TGF-β2 is crucial for the hyaluronic acid-induced epithelial-mesenchymal transition of lens epithelial cells. Purpose epithelialmesenchymal mesenchymal opacification cataract TGFβ2 TGFβ TGF β2 β (TGF)-β However TGF-β acidmediated mediated unknown Methods HLEB (HLEB3 CD44siRNA CDsiRNA siRNA CD 300 reagent CCK8 CCK 8 CCK- viability migration dichlorodihydrofluorescein dichloro dihydro fluorescein cytometer staining blotting realtime real time reaction Results 1.0 10 1 0 (1. μM 2 h CD4 medium TGFβ2mediated TGFβmediated Conclusions acidinduced induced CD44/TGFβ2 CD44TGFβ2 CDTGFβ CD44/TGF CD44/TGF-β (HLEB 30 1. (1 CD44/TGFβ CD44TGFβ CD44TGF CDTGF 3 (

Resumen Introducción : La enfermedad por arañazo de gato (EAG) es producida por Bartonella henselae y debido a que afecta principalmente niños, es poco reconocida en adultos. El cua dro evolutivo es generalmente benigno y autolimitado, aun que ocasionalmente puede haber compromiso sistémico. Métodos : Estudio observacional, descriptivo y re trospectivo realizado en un hospital de tercer nivel del conurbano bonaerense. Se incluyeron pacientes mayores de 15 años con diagnóstico de EAG en un período de 5 años (2016-2021). Resultados : Se analizaron 30 pacientes adultos, con una mediana de edad de 20.5 años (17-29), el 73% (n = 22) fueron varones. El 96% (n = 27) presentó contacto estre cho con gatos. La presentación clínica más frecuente fue adenopatías periféricas (90%); el porcentaje de compli caciones fue de 33% (n = 10). La indicación de antibio ticoterapia fue de 86.7% (n = 26), con una mediana de duración de 5 días (5-10). La evolución fue favorable en el 83% (n = 25), en el 16% (n = 5) se perdió seguimiento. Discusión : La EAG es poco reconocida en adultos; las características clínicas de este grupo etario se en cuentran poco descriptas en la literatura. Es un desafío diagnóstico debido a que su forma de presentación más frecuente es la adenopatía localizada.

Abstract Introduction : Cat-scratch disease (CSD) is caused by Bartonella henselae and it is under-recognized in adults because it mainly affects children. Clinical course is commonly benign and self-limited; occasionally, there may be systemic involvement. Methods : Case-series study carried out in a tertiary care hospital in Buenos Aires suburbs. Patients older than 15 years diagnosed with CSD over a 5-year period were included (2016-2021). Results : 30 adult patients were analyzed, with a median age of 20.5 years (IQR 17-29), 73% (n = 22) were male; 96% (n = 27) had history of exposure to cats. The most common clinical presentation of CSD was periph eral lymphadenopathy (90%), the average complication rate was 33% (n = 10), 86.7% (n = 26) received antimi crobial therapy, with a median duration of 5 days (IQR 5-10). Outcome was favorable in 83% (n = 25), 16% (n = 5) were lost to follow-up. Discussion : Clinical features of CSD in adults are poorly described in the worldwide literature. Diagnosis can be challenging because the clinical hallmark is re gional lymphadenopathy.

RESUMEN La asignatura informática ha sufrido transformaciones en su plan de estudio para estar acorde con la realidad tecnológica que se vive en la sociedad actual, ejemplo de ello es la implementación del SCRATCH como herramienta de trabajo, de ahí la importancia que tiene la preparación de los metodólogos de informática en dicha herramienta de programación (SCRATCH) para poder cumplir con sus funciones. Este trabajo tiene como objetivo ofrecer procedimientos didácticos para la preparación de los metodólogos municipales de informática de la provincia Ciego de Ávila en la enseñanza del Scratch. Para lograrlo se utilizaron diferentes métodos y tecinas: el histórico-lógico, el análisis documental y la observación al desempeño. Los resultados alcanzados permitieron la determinación de los antecedentes teóricos y metodológicos de la preparación de los metodólogos de informática para la enseñanza del SCRATCH.

ABSTRACT The computer science subject has undergone transformations in its study plan to be consistent with reality the technological reality that exists in today's society, an example of this is the implementation of Scratch as a work tool, hence the importance of the preparation of computer advisor in it´s programming tool (Scratch) to be able to fulfill their functions. This work aims to offer didactic procedures for the preparation of municipal computer advisor from Ciego de Ávila province in teaching Scratch. To achieve this, different methods and techniques were used: historical-logical, documentary analysis and performance observation. The results achieved allowed the determination of the theoretical and methodological background of the preparation of computer science advisor for teaching Scratch.

Resumo Acacia modesta (AM) e Opuntia monocantha (OM) estão distribuídas no Paquistão, Afeganistão e Índia. Ambas as plantas têm propriedades farmacológicas diferentes. Este estudo foi desenhado para avaliar o potencial anticancerígeno de Acacia modesta (AM) e Opuntia monocantha (OM). A linha celular de câncer de fígado HepG2 foi usada para avaliação da atividade anticâncer. Para a avaliação dos efeitos antiproliferativos, a viabilidade celular e a morte celular em todos os grupos de células foram avaliadas através dos ensaios de MTT, cristal violeta e azul de tripano. Para a avaliação da apoptose foi realizado ELISA de p53. Além disso, o ensaio de LDH para descobrir a capacidade das células malignas de metabolizar o piruvato em lactato e a atividade das enzimas antioxidantes (GSH, CAT e SOD) no final da HPLC foi realizado para encontrar o composto ativo de AM e OM. Citotoxicidade (MTT), ensaios de viabilidade (azul de tripano, viabilidade de cristal, análise MUSE) mostraram mais células mortas e menos células vivas nos grupos tratados com plantas com aumento da concentração. O ensaio de arranhão para o efeito anti-migratório dessas plantas mostrou que os grupos tratados não têm capacidade de cicatrizar arranhão/ferida. ELISA de p53 para apoptose celular mostrou maior liberação de p53 nos grupos tratados. Ensaio antioxidante via glutationa (GSH), superóxido dismutase (SOD), catalase (CAT) mostrou menor potencial antioxidante nos grupos de câncer tratados. O ensaio de LDH mostrou mais liberação de lactato desidrogenase nos grupos tratados em comparação com os não tratados. A análise de HPLC mostrou a presença de fitoquímicos como esteróides, alcalóides, fenóis, flavonóides, saponinas, taninos, antraquinona e aminoácidos nos extratos vegetais AM e OM. Com base em todos esses achados, pode-se concluir que os extratos etanólicos de Acacia modesta e Opuntia monocantha apresentam potencial anticancerígeno promissor.

Abstract Acacia modesta (AM) and Opuntia monocantha (OM) are distributed in Pakistan, Afghanistan and India. Both of these plants have different pharmacological properties. This study was designed to evaluate anticancer potential of Acacia modesta (AM) and Opuntia monocantha (OM). Liver cancer cell line HepG2 was used for assessment of anticancer activity. For the evaluation of anti-proliferative effects, cell viability and cell death in all groups of cells were evaluated via MTT, crystal violet and trypan blue assays. For the evaluation of apoptosis ELISA of p53 performed. Furthermore, LDH assay to find out the ability of malignant cells to metabolize pyruvate to lactate and antioxidant enzymes activity (GSH, CAT and SOD) at the end HPLC was performed to find active compound of AM and OM. Cytotoxicity (MTT), Viability assays (trypan blue, crystal viability, MUSE analysis) showed more dead, less live cells in plant treated groups with increase of concentration. Scratch assay for the anti-migratory effect of these plants showed treated groups have not ability to heal scratch/wound. ELISA of p53 for cellular apoptosis showed more release of p53 in treated groups. Antioxidant assay via glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) showed less anti-oxidative potential in treated cancer groups. LDH assay showed more lactate dehydrogenase release in treated groups compared with untreated. HPLC analysis showed the presence of phytochemicals such as steroids, alkaloids, phenols, flavonoids, saponins, tannins, anthraquinone and amino acids in AM and OM plant extracts. Based on all these findings, it can be concluded that ethanolic extracts of Acacia modesta and Opuntia monocantha have promising anti-cancer potential.

ABSTRACT To report a unique case of acute posterior multifocal placoid pigment epitheliopathy (APMPPE) in a patient with positive serology for Bartonella, presenting with ocular signs and symptoms not attributable to other diseases. A 27-year-old woman presented with decreased visual acuity in both eyes. Multimodal fundus image analysis was performed. A color fundus photograph of both eyes revealed peripapillary and macular yellow-white placoid lesions. The fundus autofluorescence of both eyes demonstrated hypo- and hyperautofluorescence of the macular lesions. Fluorescein angiography showed early-stage hypofluorescence and late staining of placoid lesions in both eyes. Spectral domain optical coherence tomography (SD-OCT) of both eyes revealed irregular elevations in the retinal pigment epithelium with the disruption of the ellipsoid zone on the topography of macular lesions. At 3 months after the treatment initiation for Bartonella infection, the placoid lesions became atrophic and hyperpigmented, and SD-OCT revealed loss of both the outer retinal layers and retinal pigment epithelium on the topography of macular lesions in both eyes. APMPPE (APMPPE diseases 27yearold yearold 27 year old performed yellowwhite yellow white hypo earlystage early stage SDOCT SD OCT (SD-OCT infection hyperpigmented 2

RESUMO Caso de epiteliopatia pigmentada placoide multifocal posterior aguda presumida em paciente com sorologia positiva para Bartonella. Paciente feminina de 27 anos apresentou diminuição da acuidade visual em ambos os olhos. Análise multimodal de imagem foi realizada. A retinografia mostrou revelou lesões placoides amarelo-esbranquiçadas nas áreas peripapilar e macular de ambos os olhos. A autofluorescência demonstrou hipo e hiperautofluorescência em ambos os olhos, na mesma topografia das lesões detectadas na retinografia. A angiofluoresceínografia mostrou hipofluorescência na fase inicial do exame e hiperfluorescência tardia das lesões placoides em ambos os olhos. A tomografia de coerência óptica de domínio espectral de ambos os olhos revelou elevações irregulares do epitélio pigmentado da retina com descontinuação da zona elipsoide na área macular. Três meses após o início do tratamento para infecção por Bartonella, as lesões placoides tornaram-se atróficas e hiperpigmentadas, e a tomografia de coerência óptica revelou perda das camadas externas da retina e do epitélio pigmentado da retina na topografia das lesões maculares em ambos os olhos. Bartonella 2 realizada amareloesbranquiçadas amarelo esbranquiçadas tornaramse tornaram se hiperpigmentadas

Abstract Objectives Nasopharyngeal carcinoma (NPC) is an aggressive epithelial cancer. The expression of miR-186 is decreased in a variety of malignancies and can promote the invasion and metastasis of cancer cells. This study aimed to explore the role and possible mechanism of miR-186 in the metastasis and epithelial-mesenchymal transformation (EMT) of NPC. Methods The expression of miR-186 in NPC tissues and cells was detected by RT-PCR. Then, miR-186 mimic was used to transfect NPC cell lines C666-1 and CNE-2, and cell activity, invasion and migration were detected by CCK8, transwell and scratch assay, respectively. The expression of EMT-related proteins was analyzed by western blotting analysis. The binding relationship between miR-186 and target gene Zinc Finger E-Box Binding Homeobox 1 (ZEB1) was confirmed by double luciferase assay. Results The expression of miR-186 in NPC was significantly decreased, and transfection of miR-186 mimic could significantly inhibit the cell activity, invasion, and migration, and regulate the protein expressions of E-cadherin, N-cadherin and vimentin in C666-1 and CNE-2 cells. Further experiments confirmed that miR-186 could directly target ZEB1 and negatively regulate its expression. In addition, ZEB1 has been confirmed to be highly expressed in NPC, and inhibition of ZEB1 could inhibit the activity, invasion, metastasis and EMT of NPC cells. And co-transfection of miR-186 mimic and si-ZEB1 could further inhibit the proliferation and metastasis of NPC. Conclusion miR-186 may inhibit the proliferation, metastasis and EMT of NPC by targeting ZEB1, and the miR-186/ZEB1 axis plays an important role in NPC. (NPC miR186 miR 186 miR-18 epithelialmesenchymal mesenchymal (EMT RTPCR. RTPCR RT PCR. PCR RT-PCR Then C6661 C C666 C666- CNE2, CNE2 CNE 2, 2 activity CCK8 CCK assay respectively EMTrelated related analysis EBox E Box ZEB (ZEB1 Ecadherin, Ecadherin cadherin, cadherin E-cadherin Ncadherin N CNE- addition cotransfection co siZEB1 siZEB si si-ZEB miR186/ZEB1 miR186ZEB1 miRZEB 186/ZEB1 miR-186/ZEB miR18 18 miR-1 C66 (ZEB miR186/ZEB miR186ZEB 186ZEB1 186/ZEB miR1 miR- C6 186ZEB

Abstract Microsatellites have been widely used to genotype individuals and to address a myriad of biological questions in many research fields for decades. However, when implementing a microsatellite marker analysis routine from scratch, various problems can arise throughout the process from DNA extraction to allele scoring, including inputting errors in the database, decreasing the reliability of results and having profound negative impacts on the derived decisions. Therefore, correctly assigning a genotype to a sample is crucial and dependent on acquiring knowledge of the technique steps, such as the chemistry of reactions, software, and data curation. This study tested two previously constructed simple sequence repeat (SSR) sets containing ten primer pairs each (ten-plex) in 1142 maize genotypes. Here, we describe the challenges faced when implementing this microsatellite-based genotyping protocol in our laboratory and possible ways to overcome them, hopefully aiding other novice research teams in this field. decades However scratch scoring database decisions Therefore steps reactions software curation SSR (SSR tenplex plex (ten-plex 114 genotypes Here microsatellitebased based them field 11 1

Abstract Objective The purpose of this study is to study the in-vitro effects of multitarget inhibitor anlotinib on hypopharyngeal cancer cell proliferation and cell migration, and the underlying mechanism, which will provide new drug choices for hypopharyngeal cancer treatment. Methods The Hypopharyngeal cancer Fadu cells were treated with anlotinib at a concentration of 0, 5, and 10 μmoL/L, respectively. Cell counting kit-8 and the colony-forming assay were used to detect the inhibition of cell proliferation. Wound-healing assay and transwell assay were used to detect the migration and invasion ability of cells. Flow cytometry was used to detect the effects of anlotinib on cell cycle and apoptosis. RT-qPCR and Western blot were used to measure gene expression levels. Results CCK-8 and colony-forming assay showed that anlotinib could significantly inhibit cell proliferative activity. Wound-healing assay and transwell assay showed that anlotinib could inhibit cell migration and scratch. These results showed that anlotinib has obvious antitumor activity. Flow cell cycle experiment showed that anlotinib could promote Fadu cell apoptosis and block the G2/M phase for inhibiting cell proliferation. In addition, anlotinib decreased the expression of HIF-1α. Conclusions Anlotinib has an excellent suppressing effect on the proliferation, migration, and invasion of hypopharyngeal cancer Fadu cells in-vitro. Moreover, it can play an anti-tumor role through blocking cell cycle G2/M and promoting apoptosis, which may be related to the decrease of HIF-1a expression. Our study would provide a potential treatment method for patients with hypopharyngeal cancer. Level of evidence: Level 3. invitro in vitro mechanism 0 5 1 μmoLL μmoL L μmoL/L respectively kit8 kit 8 kit- colonyforming colony forming Woundhealing Wound healing RTqPCR RT qPCR levels CCK8 CCK CCK- activity scratch G2M GM G2 M G addition HIF1α. HIF1α HIFα HIF 1α. 1α α HIF-1α invitro. vitro. Moreover anti tumor HIF1a HIFa 1a evidence 3

Abstract Objective This study aims to develop a compound biomaterial to achieve effective soft tissue regeneration. Methodology Compound hyaluronic acid (CHA) and liquid horizontal-platelet-rich fibrin (H-PRF) were mixed at a ratio of 1:1 to form a CHA-PRF gel. Human gingival fibroblasts (HGFs) were used in this study. The effect of CHA, H-PRF, and the CHA-PRF gel on cell viability was evaluated by CCK-8 assays. Then, the effect of CHA, H-PRF, and the CHA-PRF gel on collagen formation and deposition was evaluated by qRT‒PCR and immunofluorescence analysis. Finally, qRT‒PCR, immunofluorescence analysis, Transwell assays, and scratch wound-healing assays were performed to determine how CHA, H-PRF, and the CHA-PRF gel affect the migration of HGFs. Results The combination of CHA and H-PRF shortened the coagulation time of liquid H-PRF. Compared to the pure CHA and H-PRF group, the CHA-PRF group exhibited the highest cell proliferation at all time points, as shown by the CCK-8 assay. Col1a and FAK were expressed at the highest levels in the CHA-PRF group, as shown by qRT‒PCR. CHA and PRF could stimulate collagen formation and HGF migration, as observed by fluorescence microscopy analysis of COL1 and F-actin and Transwell and scratch healing assays. Conclusion The CHA-PRF group exhibited greater potential to promote soft tissue regeneration by inducing cell proliferation, collagen synthesis, and migration in HGFs than the pure CHA or H-PRF group. CHA-PRF can serve as a great candidate for use alone or in combination with autografts in periodontal or peri-implant soft tissue regeneration. (CHA horizontalplateletrich horizontal platelet rich HPRF H (H-PRF 11 1 1: CHAPRF (HGFs HPRF, PRF, CCK8 CCK 8 CCK- Then qRTPCR qRT PCR Finally woundhealing wound HPRF. PRF. points assay Cola Col COL Factin F actin synthesis periimplant peri implant

The application of deep learning algorithms for fault identification in wind turbine components is contingent on extensive data. Such data is often scarce, especially in the faulty category. While adversarial data augmentation helps, biases from the original data persist, and larger datasets strain computational resources. As a solution, experts are turning to transfer learning. Leveraging insights from related domains, transfer learning enables machine learning models to circumvent the exigency oftraining from scratch with extensive data. This study proposed a transfer learning strategy for fault identification in high-speed wind turbine shaft bearings. Two-dimensional matrices extracted from vibration signals sampled from the turbine bearings are employed to train ResNet50 and VGG16 convolutional neural network models with frozen weights based on transfer learning. While both models performed well on the normal test samples, they showed differing robustness when evaluated with noise-induced test samples. Contrarily, the ResNet50 had an accuracy, F-score, and training time of 82.21 %, 78.34%, and 26.3 s, respectively, while the VGG16 model had an accuracy and F-score of 95.55 % and 95.35 %, respectively, but trained for 46 s. The ResNet50 may have converged quickly due to "skip connection" in its architecture, typical of residual learning models. While the VGG16 is computationally intensive, its superior performance and resilience to noise make it suited for vibration-based defect detection in the high-speed shaft bearing, where severe background noise is prevalent.

Abstract A Hollow Cathode Plasma Enhanced Chemical Vapor Deposition (HC-PECVD) reactor was used to deposit silver doped Diamond-Like Carbon (Ag-DLC) films on Ti6Al4V alloy employing two methodologies: i) producing a silicon interlayer, using tetramethylsilane (TMS) as silicon precursor, varying the argon flow of the hollow cathode; and ii) carbonitriding the substrate. Profilometry, Raman, and Secondary Ion Mass Spectrometry (SIMS), as well as nanohardness, micro-scratch, scratch, and VDI 3198 indentation tests were used to evaluate the characteristics of the films and their adhesion on the substrates. The results demonstrated that the argon flow can be used for tuning the Ag-DLC film’s hardness, toughness, and adherence on silicon interlayers. The carbonitriding process, in turn, provided an improvement in the film toughness compared with non-carbonitrided samples. Considering the lower cost and easier handling of N2 compared to the silicon precursors commonly available (TMS, HDMSO, SiH4, etc.), the carbonitriding process proved more appropriate to improve the adhesion of the Ag-DLC films on the Ti6Al4V alloy. HCPECVD HC PECVD (HC-PECVD DiamondLike Diamond Like AgDLC Ag DLC (Ag-DLC TiAlV Ti Al V methodologies i interlayer TMS (TMS precursor cathode ii substrate Profilometry Raman SIMS, SIMS , (SIMS) nanohardness microscratch, microscratch micro scratch micro-scratch 319 substrates s hardness interlayers turn noncarbonitrided non carbonitrided samples N TMS, HDMSO SiH4 SiH etc., etc etc. etc.) (SIMS 31 3

Abstract Background: The influences of Oxycodone (OXY) combined with Paclitaxel (PTX) on breast cancer cells are unclear. The present study aimed to examine the effects of OXY combined with PTX on the proliferation, apoptosis, and migration of human breast cancer SKBR3 cells and the underlying mechanism. Methods: The proliferation, apoptosis and invasion of SKBR3 cells were assessed by CCK-8, colony formation assay, flowcytometric, Transwell assay and scratch assays, respectively. In addition, Western blotting was used to detect the expression of related proteins in these cells. The autophagic bodies were observed under a transmission electron microscope. Results: OXY (0.25, 0.5 and 1 mM) significantly inhibited the viability, colony-forming, migration, and invasion of SKBR3 cells as compared to the control group. Furthermore, OXY (0.25, 0.5 and 1 mM) markedly induced the apoptosis of SKBR3 cells and the levels of apoptosis-related proteins. In addition, OXY (0.25, 0.5 and 1 mM) and PTX inhibited the proliferation of SKBR3 cells synergistically as compared to PTX group in vitro. Moreover, OXY (0.25, 0.5 and 1 mM) significantly elevated the PTX-induced apoptosis in SKBR3 cells via downregulating the expression of N-cadherin, Becline-1 LC3-II, p-Akt and p-mTOR and upregulating E-cadherin expression. Compared with the control group, OXY (1 mM) treatment induced autophagy in SKBR3 cells. Conclusions: The present study indicates that OXY can enhance the antitumor effect of PTX on breast cancer in vitro. Hence, the combination of OXY with PTX may serve as a potential strategy for the treatment of breast cancer. Background (OXY (PTX unclear SKBR mechanism Methods CCK8, CCK8 CCK 8, 8 CCK-8 flowcytometric assays respectively addition microscope Results 0.25, 025 0 25 (0.25 05 5 0. mM viability colonyforming, colonyforming forming, forming colony-forming Furthermore apoptosisrelated vitro Moreover PTXinduced Ncadherin, Ncadherin N cadherin, cadherin N-cadherin Becline1 Becline Becline- LC3II, LC3II LCII LC3 II, II LC LC3-II pAkt p Akt pmTOR mTOR Ecadherin E ( Conclusions Hence CCK- 0.25 02 2 (0.2 0.2 (0. (0

Previous studies show that glycogen synthase kinase 3β (GSK3B) plays an important role in tumorigenesis. However, its role in cervical cancer is unclear. The present study silenced GSK3B with siRNAs and/or chemical inhibitors to determine its role in HeLa cervical cancer cell proliferation and migration as well as in xenograft tumor growth. Cell Counting Kit (CCK)-8 and 5-ethynyl-2'-deoxyuridine (EdU) assays were used to determine cell survival and proliferation. Scratch and Transwell® assays were used to evaluate cell migration. Xenograft tumors were used to evaluate the effect of GSK3B on tumor growth. Transcriptomic sequencing was used to clarify the mechanisms underlying the foregoing processes. Public databases and clinical specimens showed that GSK3B was upregulated in cervical cancer tissues and correlated with poor prognosis. In vitro experiments indicated that GSK3B inhibition reduced cell viability, proliferation, and migration. In vivo experiments demonstrated that GSK3B inhibition slowed xenograft tumor growth. Transcriptomic sequencing revealed that GSK3B inhibition modulated the phosphatidylinositol 3-carboxykinase (PI3K)/protein kinase B (Akt) and extracellular matrix (ECM)-receptor interaction signaling pathways. GSK3B inhibition decreased the protein levels of phosphorylated PI3K and Akt and the levels of mesenchymal markers but increased those of epithelial markers. An activator of the PI3K/Akt signaling pathway counteracted the suppressive effects of GSK3B inhibition on HeLa cell viability and proliferation and on PI3K/Akt signaling. Our data suggested that GSK3B regulated cervical cancer cell proliferation and migration by modulating the PI3K/Akt signaling pathway and epithelial-to-mesenchymal transition (EMT). β GSKB GSK (GSK3B tumorigenesis However unclear andor or growth CCK8 CCK 8 (CCK)- 5ethynyl2deoxyuridine ethynyldeoxyuridine 5 ethynyl 2 deoxyuridine EdU (EdU Transwell processes prognosis 3carboxykinase carboxykinase 3 PI3K/protein PI3Kprotein PIKprotein /protein PI K (Akt ECMreceptor ECM receptor pathways PIK PI3KAkt PIKAkt epithelialtomesenchymal EMT. EMT . (EMT) (CCK) Kprotein KAkt (EMT (CCK

ABSTRACT In metallic materials forming process, in general, which includes the cold rolling of stainless steels, deformation is induced by the relative movement between the material and the tool used, resulting in friction forces. However, materials behave differently during plastic deformation. In this study, microsclerometry tests were carried out to investigate the effect of different lubrication conditions on the tribological behavior of AISI 304 and AISI 430 stainless steel samples, previously hardened via cold rolling. The results of the friction coefficient and the depth of penetration of the scratches were analyzed, in addition to the scratch profile being investigated using 3D profilometry. In the microsclerometry tests, no significant differences were observed in the coefficient of friction as a function of the lubrication condition used for both steels. The coefficient of friction of the AISI 430 steel was around 23% higher than that of the AISI 304 steel due to the increase in roughness due to plastic deformation of the AISI 430 steel. The penetration depth was higher for AISI 304 steel due to the transformation of metastable austenite into martensite by deformation. Analysis of the profile of the scratches via profilometry revealed that there was no significant material removal, only displacement from the groove to the edges of the scratch. process general steels forces However study 30 43 samples analyzed D 23 removal 3 4 2

RESUMO Nos processos de conformação de materiais metálicos, como a laminação a frio de aços inoxidáveis, a deformação é induzida pelo movimento relativo entre o material e a ferramenta, resultando em forças de atrito. No entanto, os materiais apresentam comportamentos distintos durante a deformação plástica. Neste estudo, foram conduzidos ensaios de microesclerometria para investigar o efeito de diferentes condições de lubrificação no comportamento tribológico de amostras de aço inoxidável AISI 304 e AISI 430, previamente encruadas por laminação a frio. Os resultados do coeficiente de atrito e da profundidade de penetração dos riscos foram analisados, além do perfil dos riscos ter sido investigado por meio de perfilometria 3D. Nos ensaios de microesclerometria, não foram observadas diferenças significativas no coeficiente de atrito em função da condição de lubrificação para ambos os aços. O coeficiente de atrito do aço AISI 430 foi cerca de 23% maior que o do aço AISI 304, devido ao aumento de rugosidade resultante de sua deformação plástica. A profundidade de penetração foi maior para o aço AISI 304, devido à transformação da austenita metaestável em martensita durante a deformação. A análise do perfil dos riscos por perfilometria revelou que não houve remoção significativa de material, apenas deslocamento de material do sulco para as bordas do risco. metálicos inoxidáveis ferramenta entanto plástica estudo 30 analisados 3D D 43 23 risco 3 4 2

Abstract Background The newly discovered CircUBE2D2 has been shown to abnormally upregulate and promote cancer progression in a variety of cancers. The present study explored circUBE2D2 (hsa_circ_0005728) in Ovarian Cancer (OC) progression. Methods CircUBE2D2, miR-885-5p, and HMGB1 were examined by RT-qPCR or WB. SKOV-3 cell functions (including cell viability, apoptosis, migration, and invasion) were validated using the CCK-8, flow cytometry, scratch assay, and transwell assay, respectively. The direct relationship between miR-885-5p and circUBE2D2 or HMGB1 was confirmed by a dual-luciferase reporter and RNA pull-down analysis. circUBE2D2′s role in vivo tumor xenograft experiment was further probed. Results OC tissue and cell lines had higher circUBE2D2 and HMGB1 and lower miR-885-5p. Mechanically, CircUBE2D2 shared a binding relation with miR-885-5p, while miR-885-5p can directly target HMGB1. Eliminating circUBE2D2 or miR-885-5p induction inhibited OC cell activities. However, these functions were relieved by down-regulating miR-885-5p or HMGB1 induction. Furthermore, circUBE2D2 knockout reduced tumor growth. Conclusion CircUBE2D2 regulates the expression of HMGB1 by acting as a sponge of ceRNA as miR-885-5p, thereby promoting the control of OC cell proliferation and migration and inhibiting cell apoptosis. Targeting CircUBE2D2 could serve as a new potential treatment strategy for OC. CircUBED CircUBE D CircUBE2D cancers circUBED circUBE circUBE2D hsa_circ_0005728 hsacirc0005728 hsacirc hsa circ 0005728 (hsa_circ_0005728 (OC miR8855p, miR8855p miRp miR 885 5p, 5p p HMGB RTqPCR RT qPCR WB SKOV3 SKOV 3 SKOV- including viability apoptosis invasion CCK8, CCK8 CCK 8, 8 CCK-8 cytometry assay respectively dualluciferase dual luciferase pulldown pull down analysis circUBE2D2s circUBEDs s probed miR8855p. 5p. Mechanically activities However downregulating regulating Furthermore growth hsa_circ_000572 hsacirc000572 000572 (hsa_circ_000572 88 CCK- hsa_circ_00057 hsacirc00057 00057 (hsa_circ_00057 hsa_circ_0005 hsacirc0005 0005 (hsa_circ_0005 hsa_circ_000 hsacirc000 000 (hsa_circ_000 hsa_circ_00 hsacirc00 00 (hsa_circ_00 hsa_circ_0 hsacirc0 0 (hsa_circ_0 hsa_circ_ (hsa_circ_ hsa_circ (hsa_circ