PKMYT1 Promotes Epithelial-Mesenchymal Transition Process in Triple-Negative Breast Cancer by Activating Notch Signaling

ABSTRACT Background: Triple-negative breast cancer (TNBC) is a subtype of breast cancer (BC) that lacks receptors for targeted therapy. Deeper insight into the molecular mechanisms regulating TNBC metastasis is urgently needed. The epithelial-mesenchymal transition process facilitates the metastasis of neighboring epithelial tumor cells. Protein kinase, membrane-associated tyrosine/threonine 1 (PKMYT1), a member of the Wee family of protein kinases, is upregulated in BC, and its high expression predicts poor prognosis in BC patients. Notch signaling activation is a pathognomonic feature of TNBC. PKMYT1 has been found to induce EMT in non-small cell lung cancer by activating Notch signaling. However, whether PKMYT1 exerts effects on TNBC progression by regulating Notch signaling remains unknown. Objectives: The objective of this study was to investigate whether PKMYT1 exerts effects on TNBC progression by regulating Notch signaling. Methods: Fifty cases of surgically resected BC samples (tumor and adjacent non-tumor tissue samples) were collected from patients diagnosed with BC. We measured the expression of PKMYT1 in clinical samples with real-time quantitative polymerase chain reaction (RT-qPCR). For in vitro analysis, RT-qPCR and Western blotting were conducted to evaluate PKMYT1 expression in TNBC cells. Then, the viability, migration, and invasion of TNBC cells were detected by cell counting kit-8 assays, wound healing assays, and Transwell assays. The EMT event was examined by evaluating the levels of EMT-associated proteins. For in vivo analysis, xenograft models in nude mice were established to explore PKMYT1 roles. E-cadherin and Ki67 expression in xenograft models were estimated by immunohistochemistry staining. Hematoxylin and eosin staining was performed to assess tumor metastasis. The underlying mechanisms by which PKMYT1 affected the malignant phenotypes of TNBC cells were explored by Western blotting measuring the pathway-associated proteins. Results: PKMYT1 was upregulated in BC tissues and cells, and its knockdown prevented cell proliferation, migration, invasion, and EMT event in TNBC. Mechanistically, Notch signaling was inactivated by PKMYT1 depletion, and Notch activation abolished the PKMYT1 silencing-induced inhibition in the malignant phenotypes of TNBC cells. For in vivo analysis, PKMYT1 knockdown inhibited tumorigenesis and metastasis of TNBC. Conclusion: PKMYT1 promotes EMT, proliferation, migration, and invasion of TNBC cells and facilitates tumor growth and metastasis by activating Notch signaling.

Avaliação da qualidade do sono e sonolência diurna de um grupo de idosos / Assessment of sleep quality and daytime sleepiness in a group of elderly people

O objetivo do estudo foi avaliar a qualidade do sono e sonolência diurna de um grupo de idosos, verificar se há associação com prática de atividade física, presença de doença crônica, e Índice de Massa Corporal (IMC) e se há correlação com IMC, idade e qualidade de vida. Trata-se de um estudo transversal e descritivo. Para avaliação da qualidade do sono utilizou-se o Pittsburgh Sleep Quality Index (PSQI), para avaliação da sonolência diurna a Escala de Sonolência de Epworth (ESE) e para avaliação da qualidade de vida o WHOQOL-BREF. Foram avaliados 47 idosos com mediana (intervalo interquartil 25-75%) de 66 (62-70) anos de idade e IMC de 28,58 (26,21-30,44). 74,5% apresentaram sono ruim, 61,7% apresentaram Sonolência Diurna Normal e 97,8% classificados com boa qualidade de vida, com destaque para os domínios relações sociais (80%) e autoavaliação da qualidade de vida (80%). Apenas apresentou associação estatisticamente significativa a presença de qualidade de sono ruim com a prática de atividade física. Não houve associação entre presença de qualidade de sono ruim ou sonolência com IMC e presença de doença crônica. Houve uma correlação fraca, negativa e estatisticamente significativa apenas entre qualidade do sono com qualidade de vida (ρ=-0,466) e idade (ρ=-0,297). Conclui-se que os idosos apresentaram qualidade do sono ruim, sonolência diurna normal e qualidade de vida geral boa.

The objective of the study was to evaluate the quality of sleep and daytime sleepiness of a group of elderly people, checking whether there is an association with physical activity, presence of chronic disease, and Body Mass Index (BMI) and whether there is a correlation with BMI, age and quality of life. This is a cross-sectional and descriptive study. To assess sleep quality, the Pittsburgh Sleep Quality Index (PSQI) was used, the Epworth Sleepiness Scale (ESE) was used to assess daytime sleepiness, and the WHOQOL-BREF was used to assess quality of life. 47 elderly people were evaluated with a median (interquartile range 25-75%) of 66 (62-70) years of age and BMI of 28.58 (26.21-30.44). 74.5% had poor sleep, 61.7% had Normal Daytime Sleepiness and 97.8% classified as having a good quality of life, with emphasis on the domains of social relationships (80%) and self-assessment of quality of life (80%). There was only a statistically significant association between the presence of poor sleep quality and the practice of physical activity. There was no association between the presence of poor sleep quality or sleepiness with BMI and the presence of chronic disease. There was a weak, negative and statistically significant correlation only between sleep quality and quality of life (ρ=-0.466) and age (ρ=- 0.297). It is concluded that the elderly had poor sleep quality, normal daytime sleepiness and good general quality of life.

Lycophytes and ferns of an Amazonian-Cerrado ecotone in Maranhão State, Northeastern Brazil: identification key and annotated list of taxa / Licófitas e samambaias de um ecótono Amazonia-Cerrado no estado do Maranhão, Nordeste do Brasil: chave de identificação de lista anotada dos táxons

Abstract We present a key to identify and an annotated list of lycophytes and ferns of the Immediate Geographical Region of São Luís, Maranhão State, which has ecotonal characteristics of Amazonian Forest, Cerrado, and Coastal Vegetation. Specimens were collected monthly between April 2020 and September 2022 in the 13 municipalities of this region. A total of 64 species were identified, including 61 species of ferns, in 18 families and 38 genera, and three species of lycophytes, in two genera and one family. The most representative families were Pteridaceae (18 species) and Polypodiaceae (nine species). The most recorded substrate of occurrence was terrestrial with 35 species (54.6%), followed by epiphytic with 15 species (23.4%), aquatic with four species (6.2%), rupicolous with three species (4.6%), and hemiepiphytic with two species (3.1%). Five species (7.8%) had more than one life form. This study expands the knowledge about the geographic distribution of fern and lycophyte species in the central region of Northern Maranhão, in Northeastern Brazil, which until now had several sampling gaps. Maranhão has a diversity that is still poorly reported and known compared to other states in South and Southeast Brazil. Therefore, floristic surveys are essential to increase what is known about the flora, especially in areas under anthropic pressure and places with few collections, such as the ecotonal areas in Maranhão.

Resumo Apresentamos uma chave para a identificação e uma lista com detalhes das licófitas e samambaias da Região Geográfica Imediata de São Luís, Estado do Maranhão, que possui características ecotonais da Floresta Amazônica, Cerrado e Vegetação Litorânea. Os dados foram coletados mensalmente entre abril de 2020 e setembro de 2022 nos 13 municípios dessa região. Foram identificadas 64 espécies, incluindo 61 espécies de samambaias, em 18 famílias e 38 gêneros, e três espécies de licófitas, em dois gêneros e uma família. As famílias mais representativas foram Pteridaceae (18 espécies) e Polypodiaceae (nove espécies). O substrato de ocorrência mais registrado foi o terrestre com 35 espécies (54,6%), seguido do epifítico com 15 espécies (23,4%), aquático com quatro espécies (6,2%), rupícola com três espécies (4,6%) e hemiepifítico com duas espécies (3,1%). Cinco espécies (7,8%) apresentaram mais de uma forma de vida. Este estudo amplia o conhecimento sobre a distribuição geográfica de espécies de samambaias e licófitas na região central do norte do Maranhão, no Nordeste do Brasil, que até então possuía várias lacunas de amostragem. O Maranhão possui uma diversidade ainda pouco relatada e conhecida em comparação com outros estados das regiões Sul e Sudeste do Brasil. Portanto, levantamentos florísticos são essenciais para aumentar o conhecimento sobre a flora, principalmente em áreas sob pressão antrópica e locais com poucas coletas, como as áreas ecotonais do Maranhão.

Unmasking the hidden aftermath: postintensive care unit sequelae, discharge preparedness, and long-term follow-up / Desmascarando as consequências ocultas: sequelas pós-unidade de terapia intensiva, planejamento da alta e acompanhamento a longo prazo

ABSTRACT A significant portion of individuals who have experienced critical illness encounter new or exacerbated impairments in their physical, cognitive, or mental health, commonly referred to as postintensive care syndrome. Moreover, those who survive critical illness often face an increased risk of adverse consequences, including infections, major cardiovascular events, readmissions, and elevated mortality rates, during the months following hospitalization. These findings emphasize the critical necessity for effective prevention and management of long-term health deterioration in the critical care environment. Although conclusive evidence from well-designed randomized clinical trials is somewhat limited, potential interventions include strategies such as limiting sedation, early mobilization, maintaining family presence during the intensive care unit stay, implementing multicomponent transition programs (from intensive care unit to ward and from hospital to home), and offering specialized posthospital discharge follow-up. This review seeks to provide a concise summary of recent medical literature concerning long-term outcomes following critical illness and highlight potential approaches for preventing and addressing health decline in critical care survivors.

RESUMO Parcela significativa de indivíduos que enfrentaram doença crítica sofre de síndrome pós-cuidados intensivos, caracterizada por comprometimento novo ou exacerbado da função física, cognitiva ou de saúde mental. Além disso, os sobreviventes geralmente apresentam maior risco de consequências adversas, como infecção, eventos cardiovasculares maiores, reinternação e taxas de mortalidade elevadas, durante os meses após a hospitalização. Esses achados reforçam a necessidade urgente de prevenção e manejo eficazes da deterioração da saúde a longo prazo no ambiente de cuidados intensivos. Embora haja poucas evidências conclusivas de ensaios clínicos randomizados bem desenhados, potenciais intervenções incluem estratégias como limitação da sedação, mobilização precoce, presença da família durante a internação na unidade de terapia intensiva, implementação de programas de transição multidisciplinares (da unidade de terapia intensiva para a enfermaria e do hospital para o domicílio) e acompanhamento especializado após a alta hospitalar. Esta revisão objetiva fornecer um resumo conciso da literatura médica recente sobre os desfechos a longo prazo após doenças críticas e destacar potenciais abordagens para prevenir e abordar a deterioração da saúde de sobreviventes de cuidados intensivos.

Os caminhos do entrincheiramento no serviço público: uma análise longitudinal em multigrupos / Los caminos de atrincheramiento en el servicio público: un análisis longitudinal en multigrupos / The paths of entrenchment in the public service: a longitudinal analysis in multigroups

Resumo Pouco mais de uma década separa este estudo da primeira proposição acadêmica do entrincheiramento organizacional, e, até o momento, pesquisas longitudinais não foram realizadas, apesar da importância reconhecida desse método para enriquecer os estudos no campo comportamental. Este artigo objetivou identificar características do entrincheiramento organizacional entre servidores de instituições federais de ensino no Brasil, com base na avaliação longitudinal dos perfis latentes. Realizou-se uma pesquisa quantitativa e longitudinal com 1060 participantes na primeira coleta. Foram feitas análises descritivas, de comparação de médias e análise de transição latente. Os servidores da amostra apresentaram baixos níveis de entrincheiramento. Entre os principais achados, está a estabilidade dos perfis formados pelos servidores, tendo as médias mais altas sido encontradas nas dimensões "ajustamentos à posição social" e "arranjos burocráticos impessoais", o que reforça alguns achados teóricos transversais. Os resultados desta pesquisa contribuem para orientar gestores sobre a importância dos fatores internos para o entrincheiramento dos servidores e sobre como a atenção aos itens da dimensão "ajustamentos à posição social" pode favorecer o trabalho com esse vínculo. O presente estudo encontrou que, ao longo do tempo, o entrincheiramento é um vínculo estável.

Resumen Ha transcurrido poco más de una década entre la primera propuesta académica de atrincheramiento organizacional y el presente estudio y, hasta la fecha, no se ha llevado a cabo ninguna investigación longitudinal, a pesar de la reconocida importancia de este método para enriquecer los estudios en el campo del comportamiento. Esta investigación tuvo como objetivo identificar las características del atrincheramiento organizacional entre funcionarios de instituciones federales de enseñanza en Brasil, a partir de una evaluación longitudinal de perfiles latentes. Se realizó una encuesta cuantitativa y longitudinal con 1060 participantes. Se realizaron análisis descriptivos, comparación de medias y análisis de transición latente. Los funcionarios de la muestra presentaron bajos niveles de atrincheramiento. Entre los principales hallazgos están la estabilidad de los perfiles formados por los funcionarios, con las medias más altas encontradas en las dimensiones ajustes a la posición social y arreglos burocráticos impersonales, lo que refuerza algunos hallazgos teóricos transversales. Los resultados de esta investigación ayudan a orientar a los gestores sobre la importancia de los factores internos para el atrincheramiento de los funcionarios y sobre cómo la atención a los ítems de la dimensión ajustes a la posición social puede favorecer el trabajo con ese vínculo. Esta investigación constató que el atrincheramiento es un vínculo estable a lo largo del tiempo.

Abstract Little more than a decade separates this research from the first academic proposition of organizational entrenchment. To date, no longitudinal research has been carried out despite the recognized importance of this method for enriching studies in the behavioral field. This research aimed to identify characteristics of organizational entrenchment among civil servants at federal educational institutions in Brazil based on a longitudinal assessment of latent profiles. A quantitative and longitudinal survey was carried out with 1060 participants in the first collection. Descriptive analysis, comparison of means, and latent transition analysis were carried out. The civil servants in the sample showed low levels of entrenchment. Among the main findings are the stability of the profiles formed by the civil servants and the higher averages found in the dimensions of adjustments to social position and impersonal bureaucratic arrangements, reinforcing some cross-sectional theoretical findings. The results help to guide managers on the importance of internal factors for the entrenchment of civil servants and how attention to the items in the dimension of adjustments to social position can favor working with this bond. This research found that, over time, entrenchment is a stable bond.

Percepções de pessoas idosas internadas acerca da transição do cuidado do hospital para casa / Perceptions of hospitalized older adults regarding the transition of care from hospital to home

Resumo Objetivo analisar as percepções de pessoas idosas internadas acerca da transição do cuidado do hospital para casa. Método pesquisa com abordagem qualitativa, descritivo-exploratória, realizada em unidade de internação clínica de hospital público, médio porte, no interior do estado de São Paulo, Brasil. Participaram 22 pessoas idosas, por meio de entrevistas semiestruturadas, conduzidas por questões fundamentadas nos pilares do Care Transition Intervention. Posteriormente, foi realizada observação participante no local do estudo, com registro em diário de campo e transcrição das falas. Seguiram-se análise temática e sistematização dos dados por meio de software específico para análise de dados qualitativos. Resultados a média de idade dos participantes foi de 68 anos (DP±5,4); 16 (77%) possuíam uma ou mais doenças crônicas; 14 (64%) faziam uso de um ou mais medicamentos de uso contínuo. O Código Internacional de Doenças principal de internação foi covid-19. A média de permanência de internação foi de 9 dias (DP±5,7), com máximo de 31 dias e mínimo de dois dias, sendo que quatro (18%) tiveram a forma grave da doença. A partir da percepção das pessoas idosas, emergiram quatro temas de análise: problemas relacionados a medicamentos; preparo para alta hospitalar; continuidade do cuidado; e rede familiar. Conclusão na perspectiva das pessoas idosas, faz-se necessário a organização do plano de cuidados durante a internação por meio de reconciliação medicamentosa, preparo para os cuidados domiciliares, presença da família durante a internação e reconhecimento da Atenção Primária à Saúde na continuidade do cuidado durante a transição do cuidado do hospital para casa.

Abstract Objective To analyze the perceptions of older adults hospitalized regarding the transition of care from hospital to home. Method A qualitative, descriptive-exploratory research was conducted in a clinical inpatient unit of a medium-sized public hospital in the interior of São Paulo state, Brazil. Twenty-two older adults participated in the study through semistructured interviews, guided by questions based on the pillars of the Care Transitions Intervention (CTI). Subsequently, participant observation was conducted at the study site, with recording in a field diary and transcription of speeches. Thematic analysis and data systematization followed through specific software for qualitative data analysis Results The mean age of the participants was 68 years (SD±5.4); 16 (77%) had one or more chronic diseases; 14 (64%) were using one or more continuous medications. The primary International Classification of Diseases code for hospitalization was Covid-19. The average length of hospital stay was 9 days (SD±5.7), with a maximum of 31 days and a minimum of two days, with four (18%) experiencing severe disease. From the perception of older adults, four themes of analysis emerged: medication-related problems; preparation for hospital discharge; continuity of care; and family network. Conclusion From the perspective of older adults, it is necessary to organize the care plan during hospitalization through medication reconciliation, preparation for home care, family presence during hospitalization, and recognition of Primary Health Care in continuity of care during the transition from hospital to home care.

The characteristics of bone disseminated cells of 4T1 / 药学学报

Based on bone metastasis potential of mouse breast cancer 4T1 cells, the bone disseminated breast tumor cells 4T1 (B-4T1) were acquired through the screening of 6-mercaptopurine. The characteristics of B-4T1 were studied by morphological observation, proliferation assay, expression of epithelial and mesenchymal cell markers detection, transcriptome sequencing, and tumor formation experiments. The results showed that B-4T1 was round and spindle-shaped than primary 4T1 cells, and its proliferation rate was reduced, as well as epithelial cell adhesion molecule (EpCAM) and E-cadherin expression. The transcript level of N-cadherin was increased in the B-4T1, but not vimentin, indicating that B-4T1 had partial epithelial mesenchymal transition. Besides, B-4T1 had higher fatty acid metabolism and better tumor formation capacity. This study lays the experimental foundation for the basic study of metastasis in breast cancer. All animal experiments in this paper were conducted in accordance with the standards of the Animal Ethics Committee of China Pharmaceutical University.

Pathogenesis of Idiopathic Pulmonary Fibrosis and Modulating Effect of Chinese Medicine： A Review / 中国实验方剂学杂志

Idiopathic pulmonary fibrosis （IPF）， as a progressive lung disease， has a poor prognosis and no reliable and effective therapies. IPF is mainly treated by organ transplantation and administration of chemical drugs， which are ineffective and induce side effects， failing to meet the clinical needs. Therefore， developing safer and more effective drugs has become an urgent task， which necessitates clear understanding of the pathogenesis of IPF. The available studies about the pathogenesis of IPF mainly focus on macrophage polarization， epithelial-mesenchymal transition （EMT）， oxidative stress， and autophagy， while few studies systematically explain the principles and links of the pathogeneses. According to the traditional Chinese medicine theory， Qi deficiency and blood stasis and Qi-Yang deficiency are the key pathogeneses of IPF. Therefore， the Chinese medicines or compound prescriptions with the effects of replenishing Qi and activating blood， warming Yang and tonifying Qi， and eliminating stasis and resolving phlegm are often used to treat IPF. Modern pharmacological studies have shown that such medicines play a positive role in inhibiting macrophage polarization， restoring redox balance， inhibiting EMT， and regulating cell autophagy. However， few studies report how Chinese medicines regulate the pathways in the treatment of IPF. By reviewing the latest articles in this field， we elaborate on the pathogenesis of IPF and provide a comprehensive overview of the mechanism of the active ingredients or compound prescriptions of Chinese medicines in regulating IPF. Combining the pathogenesis of IPF with the modulating effects of Chinese medicines， we focus on exploring systemic treatment options for IPF， with a view to providing new ideas for the in-depth study of IPF and the research and development of related drugs.

Therapeutic Effect of Savolitinib in Patients with Stage Ⅲ/Ⅳ Non-small Cell Lung Cancer / 肿瘤防治研究

Objective To analyze therapeutic effect of savolitinib in patients with stage Ⅲ/Ⅳ non-small cell lung cancer (NSCLC). Methods A total of 95 patients with MET 14 exon (METex14) jumping mutation in stage Ⅲ/Ⅳ NSCLC were divided into a control group (47 cases) and an observation group (48 cases) through a random-number table method. The patients in the control group were treated with crizotinib, whereas those in the observation group were treated with savolitinib. The clinical efficacy and incidence of toxic side effects in both groups were evaluated through a chi-square test, and survival was evaluated through Kaplan-Meier survival analysis. Results Compared with control group (31.91% and 70.21%), the objective response rate and disease control rate of the observation group were 52.08% and 87.50%, respectively (P<0.05). According to Kaplan-Meier survival analysis, the overall survival and progression free survival rates in the observation group were higher than those in the control group (Log rank χ2=8.003, 4.528; P=0.005, 0.033). No statistically significant difference in the degree of toxic side effects was found between the groups (P>0.05). Conclusion Savolitinib can improve the efficacy of treatment for stage Ⅲ/Ⅳ METex14 skip mutation NSCLC patients, prolong survival, enhance the tolerance of patients to savolitinib, and facilitate the management of adverse reactions.

Effect Xuefu Zhuyu decoction on endothelial-to-mesenchymal transition of pulmonary artery endothelial cells and its mechanism / 中国药理学通报

Aim To investigate the effect of Xuefu Zhuyu decoction on transforming growth factor-β1(TGF-β1 ) -induced endothelial-to-mesenchymal transition (EndMT) of pulmonary microvascular endothelial cells ( PMVEC), and further analyze the mechanism related to the TGF-β1/Smad signaling pathway. Method To construct an EndMT cell model, PMVEC was treated with TGF-β1 (5 μg · L

Effect of Qingshen granules on miR-23b and PINKl/Parkin pathway in rat NRK-52E cell transdifferentiation model / 中国药理学通报

Aim To investigate the targeting mechanism of miR-23b on PINKl/Parkin pathway in transdifferentiation of NRK-52E cellsinduced by TGF-β1, and to elucidate the intervention mechanism of Qingshen granules drug-containing serum on NRK-52E cell transdifferentiation. Methods Ultra-high performance liquid chromatography ( UPLC ) fingerprinting method was used to analyze Qingshen granules. The NRK-52E transdifferentiation model induced by TGF-β1 was constructed. The NRK-52E cells were divided into simulated no-load control group, miR-23b-5p simulated group, inhibitor no-load control group, and miR-23b-5p inhibitor group, after transfection with siRNA, and the effect of miR-23b-5p on PINK1 expression was ob-served. The NRK-52E cells were then divided into normal group, TGF-(31 group, Qingshen granule group, miR-23 b-mimic group, miR-23 b-mimic group, and miR-23b-mimic + Qingshen granule group. Western blot was used to detect the expression of Pinkl, Parkin, LC3 n, Beclin-1, P62 and a-SMA proteins, and RT- PCR was used to detect the expression of miR-23 b-5p, Pinkl, Parkin, Beclin-1 and a-SMA mRNA in NRK- 52E cells. Dual-Luciferase Reporter gene experiment was used to detect the targeting relationship between miR-23b-5p and PINKL Results UPLC fingerprinting method found 11 active components in Qingshen granules. After overexpression of miR-23b-5p, the expression of PINkl mRNA significantly increased (P 0. 05 ). The experimental results showed that the expressions of miR- 23b-5p, Pinkl, Parkin, Beclin-1, LC3 II and LC3 II/ I ratio in TGF-β1 group were significantly lower than those in normal group, but the expressions of P62 and a-SMA were significantly higher than those in normal group ( P <0.05). The expressions of miR-23 b-5 p, Pinkl, Parkin, Beclin-1, LC3 II and LC3 11/ I ratio in Qingshen granule group and miR-23 b-mimic group were significantly higher than those in TGF-β1 group, and the expressions of P62 and a-SMA were significantly lower than those in TGF-β1 group (P < 0. 05 ). The performance of miR-23 b-mimic + Qingshen granule group was better than that of miR-23 b-mimic group (P < 0. 05 ). Conclusions Qingshen granules can up- regulate the expression of miR-23b-5p in NRK-52E cellsand inhibit the transdifferentiation process of NRK- 52E cells by enhancing the mitochondrial autophagy activity mediated by PINKl/Parkin pathway.

Effect of colchicine on LPS induced endothelial mesenchymal transformation and its mechanism / 中国药理学通报

Aim To investigate the effect of colchicine on lipopolysaccharide (LPS) induced endothelial to mesenchymal transition (EndMT) in human umbilical vein vascular endothelial cells (HUVECs) and its related mechanisms. Methods The EndMT model was established by treating HUVECs with LPS. Cell proliferation rate was detected by CCK-8 assay, cytotoxicity was detected by LDH assay, and the optimal drug concentration was screened. The cells were divided into the normal control group, the normal control + colchicine (10 nmol • L) group, the LPS (10 mg • L) model group, and the LPS + colchicine (10 nmol • L) group. The morphologic changes of the cells were observed under an inverted microscope, the cell migration ability was detected by Transwell assay, and the ability of tube formation was analyzed by tube formation assay. The expression of endothelial markers (CD31/ VE-cadherin) and mesenchymal cell markers (a-SMA/FSP-1) were detected by Western blot. NF-KB inhibitor was used to detect the changes in related signaling pathways. Results CCK-8 and LDH experiments showed that 10 nmol • L colchicine was the optimal concentration. LPS could induce morphological changes in HUVECs, and colchicine could reverse morphological changes in HUVECs to a certain extent. Transwell experiment showed that the migration ability of HUVECs in the LPS treatment group was significantly enhanced (P < 0. 05), and colchicine could significantly reverse this phenomenon (P < 0. 05) . Tube formation experiment showed that LPS decreased the endothelial tube formation ability of HUVECs (P < 0. 05), while colchicine treatment markedly improved LPS-induced tube formation defects (P < 0. 05) . Western blot assay showed that after colchicine co-cultured with LPS, the expression levels of CD31 and VE-cadherin significantly increased compared with the model group (P < 0. 05), while the expression levels of a-SMA and FSP-1 significantly decreased compared with the model group (P < 0. 05) . During the induction of EndMT by LPS, colchicine could inhibit the activation of the NF-KB/Snail signaling pathway. Conclusions Colchicine can effectively inhibit EndMT induced by LPS, and the mechanism may be related to the regulation of the NF-KB/Snail signaling pathway.

Effects of polydatin on the cell proliferation，migration，invasion and tumor growth of AML / 中国药房

OBJECTIVE To investigate the effects of polydatin （PD） on cell proliferation， migration， invasion and tumor growth of acute myeloid leukemia （AML）. METHODS Human AML cell KG-1 were divided into normal group， PD low-， medium- and high-concentration groups （10， 30， 60 μmol/L PD）， SQ22536 group [cyclic adenosine monophosphate （cAMP） inhibitor， 100 μmol/L]， high concentration of PD+SQ22536 group （60 μmol/L PD+100 μmol/L SQ22536）. The effects of PD on cell activity， apoptotic rate， invasion and migration ability， cAMP level， the expression of epithelial-mesenchymal transition （EMT） related proteins and protein kinase A （PKA） were investigated. Using BALB/c nude mice as subjects， a transplanted tumor model of AML nude mice was induced by subcutaneous inoculation of KG-1 cell suspension and then divided into control group， PD group， SQ22536 group and PD+SQ22536 group （with 6 mice in each group）. The effects of PD on tumor volume and mass were measured. RESULTS Compared with the normal group or control group， the cell viabilities， the number of migrating cells， the number of invasive cells， the relative expressions of vimentin and Snail as well as the tumor volume and mass were decreased significantly in PD groups， while the apoptotic rates， cAMP levels， the relative expressions of E-cadherin and PKA were significantly increased， with a dose-dependent manner （P＜0.05）. SQ22536 had opposite effects on cells and nude mice compared to PD， and could significantly reverse the anti-tumor activity of PD （P＜0.05）. CONCLUSIONS PD may inhibit the proliferation， migration， invasion and EMT process of KG-1 cells， induce apoptosis， and inhibit tumor growth， by activating the cAMP/PKA signaling pathway， thereby exerting anti-AML effects.

BMSCs promote M2 macrophage polarization to attenuate acute radiation-induced lung injury / 中国辐射卫生

Objective To investigate the therapeutic effects of bone marrow mesenchymal stem cells (BMSCs) for radiation-induced lung injury (RILI) and the underlying mechanism. Methods Forty-five healthy adult male C57BL/6 mice were randomly divided into control, model, and BMSCs groups. The model and BMSCs groups received a single irradiation dose of 20 Gy to the chest, while the control group did not receive X-ray irradiation. For the BMSCs group, an injection of 1 × 106 BMSCs cells was administered via the tail vein within 6 h after irradiation. In the 5th week, the lung tissue was taken to observe pathological changes with HE staining; examine the expression of the inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) with immunohistochemical staining; observe the polarization of macrophages with immunofluorescence staining; and measure the expression of the epithelial-mesenchymal transition markers E-cadherin, N-cadherin, and vimentin proteins by Western blot. Results After radiation, the model group developed pulmonary vasodilation and congestion with septal thickening and inflammatory cell infiltration, and these changes were markedly reduced in the BMSCs group. The model group showed significantly down-regulated expression of IL-6 and TNF-α compared with significantly increased levels in the model group (P < 0.01, P < 0.05). Treatment with BMSCs significantly increased the polarization of lung macrophages towards the M2 type, while significantly decreasing the abnormally increased N-cadherin and vimentin levels in RILI mice (P < 0.05, P < 0.01). Conclusion BMSCs have therapeutic effects for RILI mice, which may be through promoting macrophage polarization from M1 to M2.

Mechanism of TFF3-dependent TWIST1 Upregulating TRIB3 to Promote Colorectal Cancer Metastasis / 肿瘤防治研究

Objective To investigate the effects of TFF3 overexpression on the proliferation, migration, and invasion ability of colorectal cancer HT29 cells and the mechanisms involved in cancer metastasis. Methods HT29 cells were transfected with pIRES2-TFF3, and the expression levels of mRNAs and proteins related to TFF3 gene, TWIST1/TRIB3 signaling pathway, and epithelial-mesenchymal transition (EMT) were detected by qRT-PCR and Western blot. The proliferation, migration, and invasion ability of HT29 cells were detected by the CCK-8, cell scratch, and Transwell assays. Changes in cell morphology after TFF3 overexpression were observed through transmission electron microscopy. Results After the HT29 cells were transfected with pIRES2-TFF3, the expression levels of TFF3 mRNA and protein significantly increased (P<0.01); cell proliferation, migration, and invasion were significantly enhanced (P<0.01); and the expression of related genes, such as TWIST1, TRIB3, Vimentin, and Snail were significantly upregulated. By contrast, the expression of E-cadherin significantly decreased (P<0.05). Various changes in cell morphology were observed after TFF3 overexpression, such as decrease in cell junctions, loss of cilia, formation of pseudopodia, and increase in fusiform cells. Conclusion TFF3 overexpression may promote EMT in colorectal cancer cells through the Twist1/TRIB3 signaling pathway, increase their metastatic potential, and accelerate the malignant progression of colorectal cancer.

Mechanism of Medicated Serum of Buyang Huanwutang in Inhibiting Endothelial-to-mesenchymal Transition of Human Pulmonary Artery Endothelial Cells Based on Wnt1/β-catenin Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the role of the Wnt1/β-catenin signaling pathway in the intervention of medicated serum of Buyang Huanwutang （BYHWT） in endothelial-to-mesenchymal transition （EndMT） of human pulmonary artery endothelial cells （HPAECs） as well as its related mechanisms. MethodMedicated serum of BYHWT was prepared by gavage to New Zealand rabbits with a dosage of 53.36 g·kg-1·d-1 after decocting the medicine as usual. In addition， the same volume of normal saline was used to prepare blank serum. The HPAECs were cultured in vitro， and then induced by the transforming growth factor-β1 （TGF-β1） to establish the EndMT model. Five groups were established： blank group （10% blank serum）， model group （TGF-β1+10% blank serum）， low-dose BYHWT group （TGF-β1+2.5% medicated serum+7.5% blank serum）， medium-dose BYHWT group （TGF-β1+5% medicated serum+5% blank serum） and high-dose BYHWT group （TGF-β1+10% medicated serum）. Through Western blot， the expressions of Wnt1， β-catenin， and glycogen synthase kinase-3β （GSK-3β） were detected. In order to further clarify the mechanism of the Wnt1/β-catenin signaling pathway in the intervention of the medicated serum of BYHWT in inhibiting EndMT， the overexpression of β-catenin was confirmed by polymerase chain reaction after plasmid of overexpression β-catenin was constructed and transfected into the HPAECs. The HPAECs were intervened by 10% medicated serum with the optimal effect in previous studies. Then， they were divided into another five groups： the blank group （10% blank serum）， the model group （TGF-β1+10% blank serum）， the BYHWT group （TGF-β1+10% medicated serum）， the BYHWT+overexpression plasmid control group （TGF-β1+10% medicated serum+blank plasmid） and the BYHWT+β-catenin overexpression plasmid group （TGF-β1+10% medicated serum+β-catenin）. Apart from that， cell proliferation ability was detected by the methyl thiazolyl tetrazolium （MTT） method and cell migration ability by scratch assay and Transwell assay together. Immunofluorescence was adopted to detect the expressions of platelet endothelial cell adhesion molecule （PECAM-1/CD31）， vascular endothelial cadherin （VE-cadherin）， fibroblast-specific protein 1 （FSP1）， and α-smooth muscle actin （α-SMA）. ResultIn comparison to the blank group， the expressions of Wnt1 and β-catenin were significantly increased （P<0.01） while the expression of GSK-3β significantly decreased （P<0.01） in the model group. In comparison to the model group， the expressions of Wnt1 and β-catenin were significantly decreased （P<0.01） while the expression of GSK-3β was significantly increased （P<0.01） in the high-dose BYHWT group. The expression of β-catenin was significantly decreased （P<0.01） while the expression of GSK-3β was significantly increased （P<0.01） in the medium-dose BYHWT group. There was no significant difference in these indexes of the low-dose BYHWT group. In comparison to the blank group， proliferation and migration abilities were remarkably increased （P<0.01） and the immunofluorescence intensities of CD31 and VE-cadherin were decreased， while those of FSP1 and α-SMA were increased in the model group. In comparison to the model group， proliferation and migration abilities were significantly decreased （P<0.01） and the immunofluorescence intensities of CD31 and VE-cadherin were increased， while those of FSP1 and α-SMA diminished in the BYHWT group. Beyond that， the change trend of those indexes in the BYHWT+β-catenin overexpression plasmid group was consistent with that in the model group. In comparison to the BYHWT+overexpression plasmid control group， proliferation and migration abilities were significantly increased （P<0.01） and the immunofluorescence intensities of CD31 and VE-cadherin were decreased， while those of FSP1 and α-SMA were increased in the BYHWT+β-catenin overexpression plasmid group. ConclusionMedicated serum of BYHWT can inhibit EndMT of HPAECs by the Wnt1/β-catenin signaling pathway.

Targeting FAPα-positive lymph node metastatic tumor cells suppresses colorectal cancer metastasis

Lymphatic metastasis is the main metastatic route for colorectal cancer, which increases the risk of cancer recurrence and distant metastasis. The properties of the lymph node metastatic colorectal cancer (LNM-CRC) cells are poorly understood, and effective therapies are still lacking. Here, we found that hypoxia-induced fibroblast activation protein alpha (FAPα) expression in LNM-CRC cells. Gain- or loss-function experiments demonstrated that FAPα enhanced tumor cell migration, invasion, epithelial-mesenchymal transition, stemness, and lymphangiogenesis via activation of the STAT3 pathway. In addition, FAPα in tumor cells induced extracellular matrix remodeling and established an immunosuppressive environment via recruiting regulatory T cells, to promote colorectal cancer lymph node metastasis (CRCLNM). Z-GP-DAVLBH, a FAPα-activated prodrug, inhibited CRCLNM by targeting FAPα-positive LNM-CRC cells. Our study highlights the role of FAPα in tumor cells in CRCLNM and provides a potential therapeutic target and promising strategy for CRCLNM.

Jatonik polyherbal mixture induced rat liver MMPT pore opening in normal Wistar rat: In vitro and in vivo studies / 中草药·英文版

OBJECTIVE@#To assess acute toxicity, the in vitro and in vivo effects of methanol and ethyl acetate extracts (JME and JEE) of Jatonik polyherbal mixture on some mitochondria-related parameters and their effect on the activity of some liver enzymes.@*METHODS@#Acute toxicity of JME and JEE was determined using Lorke's method. In vitro and in vivo opening of the mitochondrial membrane permeability transition pore (MMPT pore) was spectrophotometrically assayed. Production of malondialdehyde (MDA) as an index of lipid peroxidation and the activity of mitochondrial ATPase was evaluated in vitro and in vivo and the effect of JME and JEE on the activity of liver enzymes such as alkaline phosphatase (ALP), aspartate and alanine aminotransferase (AST and ALT) and gamma-glutamyl transferase (GGT) was also investigated.@*RESULTS@#JME had an LD50 of 3 808 mg/kg b.w whereas JEE had an LD50 greater than 5 000 mg/kg b.w. of rats. After the rats have been fed with both extracts, a photomicrograph of a piece of liver tissue showed no apparent symptoms of toxicity. From the in vitro and in vivo studies, both extracts prompted intact mitochondria to open their MMPT pores. When compared to the control, lipid peroxide product release and ATPase activity were significantly increased (P < 0.05) in vitro and in vivo. The activities of AST, ALT, and GGT were all reduced at 50 mg/kg when treated with JME, but the activity of AST was considerably enhanced when treated with JEE (P < 0.05). The results revealed that both JME and JEE of the Jatonik polyherbal mixture had low toxicity, profound MMPTpore induction, and enhanced ATPase activity, but an increased MDA production.@*CONCLUSION@#Jatonik extracts may be a promising target for drug development in diseases where there is dysregulation of apoptosis, however, further studies are needed to better clarify the molecular mechanism involved in these phenomena.

Dendrobium Officinale Polysaccharide Alleviates Hepatic Fibrosis via Epithelial-Mesenchymal Transition / 中山大学学报(医学科学版)

ObjectiveTo investigate the effect of Dendrobium officinale polysaccharide （DOP）on CCl4-induced hepatic fibrosis（HF）and its mechanism. MethodsA total of 56 male SD rats were randomly divided into seven groups： normal group（NG），model group（MG），colchicine group（CG， 0.1 mg/kg）， Fuzheng Huayu group（FG， 0.45 g/kg），low-dose DOP group（LDG， 0.05 g/kg），middle-dose DOP group（MDG， 0.1 g/kg）and high-dose DOP group（HDG，0.2 g/kg），with 8 rats in each group. HF rat model was established by subcutaneous injection with 40% CCl4 olive oil mixture， every 3-day for 10 weeks. At the end of the sixth week， the drug groups were treated with colchicine， Fuzheng Huayu and DOP solution by gavage respectively， once a day for 4 weeks. NG and MG groups were similarly handled with an equal amount of 0.9 % normal saline. Liver histopathology was detected using hematoxylin-eosin （HE）， Masson and Sirius red staining； blood biochemistry was tested for liver function and four indicators of HF； RT-qPCR and Western Blot were used to measure the expression of α-SMA， Col-I， E-cadherin， and ZEB1 genes and proteins in the liver tissues of rats， respectively. ResultsHE， Masson， and Sirius red staining showed that the liver tissue of MG rats had typical pathologic features of HF， and the degree of HF was alleviated in LDG， MDG， and HDG rats， respectively. Liver function test results showed that the serum AST， TBIL， and AKP levels were significantly lower in LDG， MDG， and HDG， compared with those of the MG （P < 0.05 or < 0.01）. Meanwhile， ALT levels in serum deceased remarkably except in LDG （P < 0.05 or < 0.01）. The four results of HF showed that the serum HA， LN， PC-Ⅲ， and COL-Ⅳ levels in LDG， MDG， and HDG rats were significantly decreased compared with those of the MG （P < 0.05 or < 0.01）. The relative expressions of α-SMA， COL-I， and ZEB1 genes and proteins were significantly decreased in the liver tissues of LDG， MDG， and HDG （P < 0.05 or < 0.01）， and the relative expression of E-cadherin gene and protein increased （P < 0.05 or < 0.01）. In addition， the expressions of HA， α-SMA， COL-I， ZEB1 and E-cadherin were dependent on the dose of DOP. ConclusionDOP alleviated the degree of CCl4 induced HF in rats by inhibiting the epithelial-mesenchymal transition in liver tissue.

Correlation Between Wnt5a Expression and Vasculogenic Mimicry in Prostate Cancer Tissues / 肿瘤防治研究

Objective To investigate the correlation of Wnt5a expression and vasculogenic mimicry (VM) in prostate cancer tissues, and analyze their relationships with cancer stem cells (CSCs) characteristics and epithelial–mesenchymal transition (EMT). Methods Immunohistochemistry was conducted to detect the expression of Wnt5a in 50 prostate cancer tissues and 50 benign prostatic hyperplasia tissues. The expression levels of CD133, vimentin, and E-cadherin were detected in the prostate cancer tissues, and CD34/PAS double staining was used to detect VM structures. We analyzed the difference in Wnt5a level between prostate cancer and benign prostatic hyperplasia tissues, the clinical significance of Wnt5a and VM, the relationship of Wnt5a expression and VM, and the relationships of Wnt5a expression and VM with CD133, Vimentin, E-cadherin. Results The expression of Wnt5a was significantly higher in prostate cancer tissues than in benign prostatic hyperplasia (P < 0.05). A positive correlation was observed between Wnt5a expression and VM (P < 0.05). The expression levels of Wnt5a and VM were positively correlated with those of CD133 and vimentin (P < 0.05). Wnt5a expression and VM were positively correlated with Gleason score, vas deferens invasion and lymphatic metastasis (P < 0.05) of prostate cancer, and VM was also positively correlated with T stage of prostate cancer (P < 0.05). Conclusion The expression level of Wnt5a in prostate cancer tissues is elevated and positively related with VM formation. Wnt5a expression and VM are correlated with cancer stem cells characteristics and the expression of epithelial–mesenchymal transition marker proteins.