Single-cell RNA sequencing reveals the mediatory role of cancer-associated fibroblast PTN in hepatitis B virus cirrhosis-HCC progression.

BACKGROUND: Cancer-associated fibroblasts (CAFs) are essential stromal components in the tumor microenvironment of hepatocellular carcinoma (HCC). Hepatitis B virus (HBV) infection induces pathological changes such as liver fibrosis/cirrhosis and HCC. The aim of this research was to explore the novel mediators of CAFs to modulate HBV cirrhosis-HCC progression. METHODS: The single-cell transcriptome data of HCC were divided into subsets, and the significant subset related to fibrotic cells, along with biological function, and clinical information of HCC was revealed by integrated data analyses. The cell communication, cells communicated weight analysis of signaling pathways, and key genes in signaling pathways analysis of significant CAFs subclasses were conducted to discover the novel gene of CAFs. Bioinformatics, vitro and HBV transfection assays were used to verify the novel gene is an important target for promoting the progression HBV cirrhosis-HCC progression. RESULTS: Fibroblasts derived from HCC single-cell data could be separated into three cell subclasses (CAF0-2), of which CAF2 was associated with the HCC clinical information. Fibroblasts have opposite developmental trajectories to immune B cells and CD8 + T cells. CAF0-2 had strong interaction with B cells and CD8 + T cells, especially CAF2 had the highest interaction frequency and weight with B cells and CD8 + T cells. Moreover, PTN participated in CAF2-related pathways involved in the regulation of cell communication, and the interactions among CAF2 and PTN contributed the most to B cells and CD8 + T cells. Furthermore, the genes of PTN, SDC1, and NCL from PTN signaling were highest expression in CAF2, B cells, and CD8 + T cells, respectively, and the interaction of PTN- SDC1 and PTN- NCL contributed most to the interaction of CAF2- B cells and CAF2-CD8 + T cells. Bioinformatics and vitro experiments confirm PTN was upregulated in HCC and promoted the proliferation of tumor cells, and HBV infection could initiate PTN to perform cirrhosis-HCC progression. CONCLUSION: Our findings revealed CAF was associated with hepatocarcinogenesis, and the functional importance of B cells and CD8 + T cells in modulating CAF in HCC. Importantly, PTN maybe a novel mediator of CAF to mediate HBV cirrhosis-HCC progression.

Osmotic pressure induces translocation of aquaporin-8 by P38 and JNK MAPK signaling pathways in patients with functional constipation.

BACKGROUND: Aquaporins (AQPs) maintain fluid homeostasis in the colon. The role of colonic AQPs in the pathophysiology of functional constipation (FC) remains largely unknown. AIM: To explore variations in aquaporins and investigate their underlying mechanisms. METHODS: Colonic biopsies were collected from patients with FC and healthy controls. The expression and localization of AQPs were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR), western blot analysis, and immunofluorescence assays. Furthermore, osmotic pressure-induced cell model was used in vitro to investigate the potential relationship between AQP8 and osmotic pressure, and to reveal the underlying mechanisms. RESULTS: Upregulation of AQP3 and AQP8, and downregulation of AQP1, AQP7, AQP9, AQP10, and AQP11 were observed in the patients with functional constipation. Furthermore, cellular translocation of AQP8 from the cytoplasm to the plasma membrane was observed in patients with FC. Mechanistically, the increase in osmotic pressure could activate the Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) signaling pathways, and subsequently promote the upregulation and translocation of AQP8. CONCLUSION: Upregulation of AQP8 and AQP3, and translocation of AQP8 were observed in colon biopsies from patients with FC. The p38 and JNK MAPK signaling pathways are involved in the regulation of osmotic pressure-induced AQP8 variation.

Evaluation of high levels of sports activity and the beneficial effect on postprandial blood glucose profiles / Avaliação dos altos níveis de atividade esportiva e o efeito benéfico nos perfis de glicemia pós-prandial / Evaluación de los niveles elevados de actividad deportiva y el efecto positivo en los perfiles de glucemia postprandial

ABSTRACT Introduction: Hyperglycemia is the principal characteristic component of type 2 diabetes. High blood glucose concentrations for long periods can be countered with postprandial exercise by increasing glucose retention involuntary muscles. However, no research is present on the relationship between exercise time and glucose levels. Objective: This study evaluates the relationship between sports activity and postprandial glycemia levels. Methodology: Forty-five individuals were included in the study, 10 males and 35 females with an age of 27.11±2.8 years; a body fat percentage of 25.02% ±5.04%; and a body mass index of 22.74±4.55 kg/m2. Participants were included via WhatsApp for daily information on postprandial activity levels. WhatsApp messages were forwarded to a total of 2,500 people at different colleges and universities. Out of the total 60 active people (2.40%) who responded, 45 individuals participated in the study. They were divided into three categories based on self-reported postprandial activity: not very active (15), quite active (15), highly active (15). All active individuals completed an oral glucose intake test with blood samples obtained for evaluation at 15, 30, 45, 60, 90, and 120 minutes post-rest. On a gender basis, the groups could not be associated (P =.057). Results: All active groups showed a remarkable effect on blood glucose level at one hour (P =.031). A mean increase in blood glucose level in the first hour of 1.50 mmol/L was observed for every extra 1.0 mmol/L of standard glycemic amount, on average, women had a higher blood glucose amount of 1.35 mmol/L than men. Conclusion: It can be concluded that a high amount of postprandial activity generates a good outcome on glycemic parameters. Evidence Level II; Therapeutic Studies - Investigating the results.

RESUMO Introdução: A hiperglicemia é o principal componente característico na diabetes tipo 2. Altas concentrações de glicose por longos períodos podem ser combatidas com o exercício pós-prandial, aumentando a retenção de glicose nos músculos voluntários. Porém, ainda não há estudos sobre a relação entre o tempo de exercício e os níveis de glicose. Objetivo: Este estudo tem como objetivo avaliar a relação entre a atividade esportiva e os dados temporais de glicemia pós-prandial. Metodologia: Foram incluídos 45 indivíduos no estudo, sendo 10 do sexo masculino e 35 do sexo feminino com idade de 27,11± 2,8 anos; percentual de gordura corporal de 25,02% ±5,04%; e índice de massa corporal de 22,74±4,55 kg/m2. Os participantes foram incluídos via WhatsApp para obter informações diárias sobre os níveis de atividade pós-prandial. As mensagens de WhatsApp foram encaminhadas para um total de 2.500 pessoas em diferentes faculdades e universidades. No total de 60 pessoas ativas (2,40%) que responderam, participaram do estudo 45 indivíduos. Eles foram divididos em três categorias com base na atividade pós-prandial autorrelatada: pouco ativos (15), bastante ativos (15), altamente ativos (15). Todos os indivíduos ativos finalizaram um teste de ingestão de glicose oral com amostras de sangue obtidas para avaliação em 15, 30, 45, 60, 90 e 120 minutos pós-repouso. Na base de gênero, os grupos não puderam ser associados (P =.057). Resultados: Todos os grupos ativos revelaram um efeito notável do nível de glicose no sangue em uma hora (P =.031). Foi observado um aumento médio no nível de glicemia na primeira hora de 1,50 mmol/L para cada 1,0 mmol/L extra de quantidade glicêmica padrão, em média, as mulheres tiveram uma quantidade glicêmica no sangue de 1,35 mmol/L superior aos homens. Conclusão: Conclui-se que a alta quantidade de atividade pós-prandial gera um bom desfecho nos parâmetros glicêmicos. Nível de evidência II; Estudos Terapêuticos - Investigação de Resultados.

RESUMEN Introducción: La hiperglucemia es el principal componente característico de la diabetes de tipo 2. Las concentraciones elevadas de glucosa durante largos periodos pueden combatirse con el ejercicio postprandial, aumentando la retención de glucosa en los músculos voluntarios. Sin embargo, todavía no hay estudios sobre la relación entre el tiempo de ejercicio y los niveles de glucosa. Objetivo: Este estudio pretende evaluar la relación entre la actividad deportiva y los datos de glicemia postprandial. Metodología: Se incluyeron 45 individuos en el estudio, siendo 10 hombres y 35 mujeres con una edad de 27,11±2,8 años; un porcentaje de grasa corporal de 25,02% ±5,04%; y un índice de masa corporal de 22,74±4,55 kg/m2. Se inscribió a los participantes a través de WhatsApp para obtener información diaria sobre los niveles de actividad postprandial. Se enviaron mensajes de WhatsApp a un total de 2.500 personas de diferentes colegios y universidades. Del total de 60 personas activas (2,40%) que respondieron, 45 individuos participaron en el estudio. Fueron divididos en tres categorías basadas en la actividad postprandial auto declarada: poco activos (15), bastante activos (15), muy activos (15). Todos los individuos activos completaron una prueba de ingesta de glucosa oral con muestras de sangre obtenidas para su evaluación a los 15, 30, 45, 60, 90 y 120 minutos después del reposo. En lo que respecta al género, los grupos no pudieron asociarse (P = 0,057). Resultados: Todos los grupos activos revelaron un efecto notable del nivel de glucosa en la sangre a una hora (P = 0,031). Se observó un aumento medio del nivel de glucosa en la sangre en la primera hora de 1,50 mmol/L por cada 1,0 mmol/L adicional de la cantidad de glucemia estándar; por término medio, las mujeres tuvieron una cantidad de glucosa en la sangre más alta de 1,35 mmol/L que los hombres. Conclusión: Se concluye que la elevada actividad postprandial genera un buen resultado en los parámetros glucémicos. Nivel de evidencia II; Estudios terapéuticos - Investigación de resultados.

Refinement of saliva mi-rna biomarkers for sport-related concussion / Refinamento de biomarcadores de saliva mi-rna para concussão relacionada ao esporte / Refinamiento de los biomarcadores de mi-arn de la saliva para la conmoción encefálica relacionada con el deporte

ABSTRACT Introduction: The changes in brain structure caused by a sports-related concussion may initially be indistinguishable, however, the irreversible deleterious effects are noted in the long term. An early diagnosis may provide the patient with a better recovery chance and increased survival. For this purpose, this paper studies the feasibility of a diagnosis for concussion by microRNA (mi-RNA) biomarkers contained in the saliva of athletes. Objective: Verify whether salivary miRNAs could be considered good biomarkers for sports concussion. Methodology: Salivary mi-RNA levels were determined from 120 saliva samples of 120 players. There were 43 with a diagnosis of concussion and 77 without a diagnosis of concussion. Samples from players with a concussion were collected 30 minutes prior to activity, samples from individuals who did not engage in physical activity were also compared. Results: On the evaluation of 30 miRNA from individuals with a concussion between contact and non-contact sports there was high detection reliability(P<.05). Both miR-532-5p and miR-182-5p showed reduced amounts of physical activity. The miRNA-532-5p and miRNA-182-5p show significant results among 43 subjects from pre-exercise to post-exercise. The miRNA-4510 showed a significant result (p < 0.05) between contact and non-contact sport types. The amount of miRNA-4510 expanded in 20 individuals in the contact sport at post-exercise but remained normal in the non-contact sports group. Conclusion: The salivary miRNAs are reliable biomarkers for concussion. Evidence Level II; Therapeutic Studies - Investigating the results.

RESUMO Introdução: As alterações da estrutura cerebral provocadas por uma concussão relacionada ao esporte podem ser inicialmente indistinguíveis, porém os efeitos deletérios irreversíveis são notados a longo prazo. Um diagnóstico precoce poderá fornecer ao paciente uma chance maior de recuperação e aumento de sobrevida. Para tanto, estuda-se a viabilidade de um diagnóstico de concussão por biomarcadores de micro RNA (mi-RNA) contidos na saliva de esportistas. Objetivo: verificar se os miRNAs salivares são biomarcadores confiáveis para concussão esportiva. Metodologia: Os níveis de mi-RNA salivares foram determinados a partir de 120 amostras de saliva de 120 jogadores. Haviam 43 com diagnóstico de concussão e 77 sem diagnóstico de concussão. Amostras de jogadores com concussão foram coletadas 30 minutos antes da atividade, amostras de indivíduos que não praticaram atividade física também foram comparadas. Resultados: Na avaliação de 30 mi-RNA de indivíduos com concussão entre esportes de contato e sem contato houve grande confiabilidade de detecção(P<,05). Tanto o miR-532-5p quanto o miR-182-5p mostraram quantidades reduzidas na atividade física. O miRNA-532-5p e o miRNA-182-5p mostram resultados significativos entre 43 indivíduos desde o pré-exercício até o pós-exercício. O miRNA-4510 mostrou um resultado significativo (p < 0,05) entre os tipos de esporte com contato e sem contato. A quantidade de mi-RNA-4510 expandiu-se em 20 pessoas no esporte com contato no pós-exercício, mas permaneceu normal no grupo de esporte sem contato. Conclusão: Conclui-se que os miRNAs salivares são biomarcadores confiáveis para concussão. Nível de evidência II; Estudos Terapêuticos - Investigação de Resultados.

RESUMEN Introducción: Los cambios en la estructura cerebral causados por una conmoción encefálica relacionada con el deporte pueden ser inicialmente indistintos, sin embargo, los efectos deletéreos irreversibles se manifiestan a largo plazo. Un diagnóstico anticipado puede proporcionar al paciente una mayor posibilidad de recuperación y una mayor supervivencia. Por lo tanto, se estudia la viabilidad de un diagnóstico de conmoción encefálica mediante los biomarcadores de microARN (mi-ARN) contenidos en la saliva de los deportistas. Objetivo: verificar si los miARN salivales son biomarcadores fiables para la conmoción encefálica deportiva. Metodología: Se determinaron los niveles de mi-ARN salival a partir de 120 muestras de saliva de 120 jugadores. Había 43 con diagnóstico de conmoción encefálica y 77 sin diagnóstico de conmoción encefálica. Las muestras de los jugadores con conmoción encefálica se recogieron 30 minutos antes de la actividad, también se compararon las muestras de los individuos que no practicaban actividad física. Resultados: En la evaluación de 30 miRNA de individuos con conmoción encefálica entre deportes de contacto y sin contacto hubo una alta fiabilidad de detección (P<.05). Tanto miR-532-5p como miR-182-5p mostraron cantidades reducidas en la actividad física. El miRNA-532-5p y el miRNA-182-5p muestran resultados significativos entre los 43 sujetos desde antes del ejercicio hasta después del mismo. El miRNA-4510 mostró un resultado significativo (p < 0,05) entre los tipos de deporte de contacto y de no contacto. La cantidad de miARN-4510 se expandió en 20 individuos en el deporte de contacto en el momento posterior al ejercicio, pero se mantuvo normal en el grupo de deporte sin contacto. Conclusión: Llegamos a la conclusión de que los miARN salivales son biomarcadores fiables para la conmoción encefálica. Nivel de evidencia II; Estudios terapéuticos - Investigación de resultados.

Effect of altered gut microbiota on visceral hypersensitivity of postinfectious irritable bowel syndrome mice.

OBJECTIVE: Irritable bowel syndrome (IBS) is a common functional bowel disorder characterized with visceral hypersensitivity. Previous studies indicated gut microbiota alteration associated short-chain fatty acids (SCFAs) dysregulation is associated with IBS development. The aim of the study is to explore the potential role of microbiota dysbiosis mediated visceral hypersensitivity in postinfectious-IBS (PI-IBS) mouse model. METHODS: Four-week-old NIH mice were randomly allocated into four groups: control mice, PI-IBS mice, PI-IBS mice co-housing with normal mice, and PI-IBS mice were administrated with a cocktail of antibiotics. Trichinella spiralis infection established PI-IBS mouse model. Microbiota in cecal contents and feces were analyzed by 16S rDNA sequencing. SCFAs were detected by gas chromatography. 5-hydroxytryptamine (5-HT) was evaluated by ELISA, and N-methyl-D-aspartate receptors (NMDARs) were examined by western blot. Visceral sensitivity was determined by abdominal withdrawal reflex in response to colorectal distention. RESULTS: Increased SCFAs were observed in cecal contents and feces in PI-IBS mice accompanied with higher 5-HT and NMDAR subunits expressions in ileum and colon. Visceral hypersensitivity was observed in PI-IBS mice compared to control mice. When administrated with antibiotics cocktails and co-housing with normal mice, PI-IBS mice showed decreased SCFAs, 5-HT, NMDAR subunits expressions, and improved visceral hypersensitivity. CONCLUSION: Gut microbiota alteration induced increased SCFAs, 5-HT and NMDAR subunits expressions were associated with visceral hypersensitivity in PI-IBS mice. The critical role of gut microbiota in improving visceral hypersensitivity was further identified by treatment of antibiotics cocktail and co-housing.

miR-1275 Inhibits Human Omental Adipose-Derived Stem Cells Differentiation Toward the Beige Phenotype via PRDM16.

Beige adipocytes have recently attracted attention for their potential as new therapeutic targets in the management of obesity and related metabolic disorders. MicroRNAs (miRNAs) have been reported as transcriptional regulators or biomarkers of brown and beige adipogenesis. Nevertheless, the effects of miRNAs involved in beige differentiation of human visceral adipocytes remain to be investigated. In this study, microarray screening showed that miR-1275 was significantly decreased during the differentiation of beige adipocytes induced by human omental adipose-derived stem cells (hASCs). Overexpression of miR-1275 suppressed the "brown-like" differentiation of hASCs by inhibiting the key transcriptional factor PR domain containing 16 (PRDM16) without affecting the proliferation. Adipogenesis and mitochondrial biogenesis of beige adipocytes derived from hASCs were impaired by miR-1275 overexpression. The regulatory effect of miR-1275 was determined by direct binding to the 3'-untranslated region of PRDM16, which was demonstrated by a dual-luciferase assay. Taken together, this study identified miR-1275 as a negative regulator of beige cell development in hASCs by inhibiting PRDM16. Thus, miR-1275 might be a potential target in the management of visceral obesity and related metabolic diseases.

Transcutaneous Electrical Acustimulation Improves Irritable Bowel Syndrome With Constipation by Accelerating Colon Transit and Reducing Rectal Sensation Using Autonomic Mechanisms.

INTRODUCTION: Slow colon transit and visceral hypersensitivity are recognized as major pathophysiological mechanisms in irritable bowel syndrome with constipation (IBS-C). However, there is a lack of therapies targeting both abdominal pain and colonic motility. This study was designed to investigate the long-term effects and possible mechanisms of transcutaneous electrical acustimulation (TEA) in patients with IBS-C. METHODS: Fifty-two patients with IBS-C were randomized into 2 groups: daily TEA for 4 weeks (n = 26) and daily sham-TEA for 4 weeks (n = 26). The number of complete spontaneous bowel movements per week (CSBMs/week, primary outcome), Irritable Bowel Syndrome Severity Scoring System, Patient Assessment of Constipation Quality of Life, visual analog scale (VAS) pain score, colonic transit time, and anorectal physiology were evaluated before treatment and at the end of the treatment. Colonic transit was assessed with radiopaque markers. Electrocardiograms were recorded for assessing autonomic functions. RESULTS: (i) TEA improved constipation and abdominal pain. After the treatment, the number of CSBMs/week during the last week in the TEA group was higher than that in the sham-TEA group (3.5 ± 1.6 vs 2.3 ± 0.6, P = 0.002). Similar effects were also noted in the visual analog scale pain score ( P = 0.002) and Irritable Bowel Syndrome Severity Scoring System score ( P = 0.025). In addition, there was a significant improvement in the quality of life of patients with constipation. The Patient Assessment of Constipation Quality of Life total score was significantly decreased in the TEA group ( P = 0.004). (ii) Compared with sham-TEA, TEA improved colon transit ( P = 0.002) and increased the threshold of rectal sensation (desire to defecate, P = 0.004; maximum tolerability, P < 0.001). (iii) TEA increased vagal activity, compared with sham-TEA ( P < 0.05); at the end of the treatment, the vagal activity was significantly correlated with colon transit and the CSBMs/week. DISCUSSION: TEA improves constipation and symptoms of IBS by accelerating colon transit and reducing rectal sensation, possibly mediated by using the autonomic mechanisms.

Construction and validation of a prognostic risk model for breast cancer based on protein expression.

Breast cancer (BRCA) is the primary cause of mortality among females globally. The combination of advanced genomic analysis with proteomics characterization to construct a protein prognostic model will help to screen effective biomarkers and find new therapeutic directions. This study obtained proteomics data from The Cancer Proteome Atlas (TCPA) dataset and clinical data from The Cancer Genome Atlas (TCGA) dataset. Kaplan-Meier and Cox regression analyses were used to construct a prognostic risk model, which was consisted of 6 proteins (CASPASE7CLEAVEDD198, NFKBP65-pS536, PCADHERIN, P27, X4EBP1-pT70, and EIF4G). Based on risk curves, survival curves, receiver operating characteristic curves, and independent prognostic analysis, the protein prognostic model could be viewed as an independent factor to accurately predict the survival time of BRCA patients. We further validated that this prognostic model had good predictive performance in the GSE88770 dataset. The expression of 6 proteins was significantly associated with the overall survival of BRCA patients. The 6 proteins and encoding genes were differentially expressed in normal and primary tumor tissues and in different BRCA stages. In addition, we verified the expression of 3 differential proteins by immunohistochemistry and found that CDH3 and EIF4G1 were significantly higher in breast cancer tissues. Functional enrichment analysis indicated that the 6 genes were mainly related to the HIF-1 signaling pathway and the PI3K-AKT signaling pathway. This study suggested that the prognosis-related proteins might serve as new biomarkers for BRCA diagnosis, and that the risk model could be used to predict the prognosis of BRCA patients.

Lamina propria interleukin 17 A aggravates natural killer T-cell activation in autoimmune hepatitis.

Autoimmune hepatitis is an interface hepatitis characterized by the progressive destruction of the liver parenchyma, the cause of which is still obscure. Interleukin (IL)-17A is a major driver of autoimmunity, which can be produced by innate immune cells against several intracellular pathogens. Here, we investigated the involvement of IL-17A in a mice model of immune-mediated hepatitis with the intestine exposed to Salmonella typhimurium. Our results showed more severe Concanavalin (Con) A-induced liver injury and gut microbiome dysbiosis when the mice were treated with a gavage of S. typhimurium. Then, the natural killer (NK) T cells were overactivated by the accumulated IL-17A in the liver in the Con A and S. typhimurium administration group. IL-17A could activate NKT cells by inducing CD178 expression via IL-4/STAT6 signaling. Furthermore, via the portal tract, the laminae propria mucosal-associated invariant T (MAIT)-cell-derived IL-17A could be the original driver of NKT cell overactivation in intragastric administration of S. typhimurium and Con A injection. In IL-17A-deficient mice, Con A-induced liver injury and NKT cell activation were alleviated. However, when AAV-sh-mIL-17a was used to specifically knock down IL-17A in liver, it seemed that hepatic IL-17a knock down did not significantly influence the liver injury. Our results suggested that, under Con A-induction, laminae propria MAIT-derived IL-17A activated hepatic NKT, and this axis could be a therapeutic target in autoimmune liver disease.

Secreted phosphoprotein 1 as a potential prognostic and immunotherapy biomarker in multiple human cancers.

Secreted phosphoprotein 1 (SPP1) is involved in immune regulation, cell survival, and tumor progression. Studies have demonstrated that SPP1 plays an important role in certain individual tumors. However, the expression profile and oncogenic features of SPP1 in diverse cancers are remaining unknown. Therefore, we performed a comprehensive analysis using The Cancer Genome Atlas (TCGA) database. Raw data of 33 cancer types were download from the University of California Santa Cruz (UCSC) Xena website. The expression of SPP1 and its relationship with tumor prognosis, immune invasion, tumor microenvironment, and immunotherapy were analyzed using the R language. The function analysis was conducted using Gene Set Enrichment Analysis (GSEA). The oncogenic features of SPP1 was validated by wound-healing assay and EdU staining assay. SPP1 highly expressed in most cancers. The expression of SPP1 was significant related to prognosis, tumor mutation burden (TMB), microsatellite instability (MSI), and immune checkpoint genes, suggested that SPP1 plays an essential role in the tumor immune microenvironment and immune cell infiltration. The immune/stromal scores correlated positively with the SPP1 expression, and the relationship was affected by tumor heterogeneity and immunotherapy. In addition, SPP1 could predict the response of tumor immunotherapy. Functional analysis revealed the SPP1-associated terms and pathways. Finally, SPP1 significantly elevated cell proliferation and migration in A549, Huh7, HT-29, A2780 tumor cell lines. In conclusion, this study indicated that SPP1 involved in tumorigenesis, tumor progression, and regulated tumor immune microenvironment, revealing SPP1 might be a potential target for evaluating prognosis and immunotherapy in multiple cancers.

Effect of 8-week high-intensity intermittent running exercise and weight training on muscle and DNA damage in male and female ski major college students.

This study assessed changes in blood muscle damage indicators and DNA damage indicators in lymph and urine after 8 weeks of high-intensity intermittent running and weight training in male and female college students majoring in skiing. This study aimed to find an effective training method by investigating differences in the effectiveness between men and women. A total of 20 male and female ski major college students conducted short-term high-intensity intermittent running and weight training in the morning and afternoon, respectively, 3 days a week for 8 weeks for 24 times in total. After 8 weeks of high-intensity intermittent running and weight training, changes in DNA damage indicators in the lymph and urine and muscle damage indicators in the blood were analyzed. The creatine kinase level significantly differed at rest pre-graded exercise testing (GXT) and 60 min of recovery post-GXT after training from that before training between the male and female groups. Although lactate dehydrogenase (LDH) levels decreased in both groups over time, no significant differences in LDH were found between the two groups. Second, DNA 8-hydroxydeoxyguanosine (8-OHdG) in the lymph was significantly different between the two groups at rest pre-GXT and 60 min of recovery post-GXT. 8-OHdG in the urine was significantly lower in the female group only at 60 min of recovery post-GXT. Partial sex differences were found in the reduction of muscle damage and DNA damage after 8 weeks of high-intensity intermittent running and weight training.

In Vitro Study of Human Immune Responses to Hyaluronic Acid Hydrogels, Recombinant Spidroins and Human Neural Progenitor Cells of Relevance to Spinal Cord Injury Repair.

Scaffolds of recombinant spider silk protein (spidroin) and hyaluronic acid (HA) hydrogel hold promise in combination with cell therapy for spinal cord injury. However, little is known concerning the human immune response to these biomaterials and grafted human neural stem/progenitor cells (hNPCs). Here, we analyzed short- and long-term in vitro activation of immune cells in human peripheral blood mononuclear cells (hPBMCs) cultured with/without recombinant spidroins, HA hydrogels, and/or allogeneic hNPCs to assess potential host-donor interactions. Viability, proliferation and phenotype of hPBMCs were analyzed using NucleoCounter and flow cytometry. hPBMC viability was confirmed after exposure to the different biomaterials. Short-term (15 h) co-cultures of hPBMCs with spidroins, but not with HA hydrogel, resulted in a significant increase in the proportion of activated CD69+ CD4+ T cells, CD8+ T cells, B cells and NK cells, which likely was caused by residual endotoxins from the Escherichia coli expression system. The observed spidroin-induced hPBMC activation was not altered by hNPCs. It is resource-effective to evaluate human compatibility of novel biomaterials early in development of the production process to, when necessary, make alterations to minimize rejection risk. Here, we present a method to evaluate biomaterials and hPBMC compatibility in conjunction with allogeneic human cells.

lncRNA-HEIM Facilitated Liver Fibrosis by Up-Regulating TGF-ß Expression in Long-Term Outcome of Chronic Hepatitis B.

Background: Chronic liver fibrosis is an inevitable stage for the development of patients with chronic hepatitis B (CHB). However, anti-fibrotic therapies have been unsuccessful so far. The biological functions and molecular mechanisms of long non-coding RNAs (lncRNAs) in the host immune system during chronic hepatitis B virus (HBV) infection, especially in fibrosis, are still largely unknown. Method: The total RNA of peripheral blood mononuclear cells (PBMCs) from asymptomatic carriers (ASCs) or CHB receiving at least 8 years of anti-viral treatments was analyzed using Arraystar microarray and validated via quantitative real-time PCR (qRT-PCR). Correlation analysis was conducted based on correlation coefficients, Clusterprofile, and RNA Interactome Database (RAID). The functions of lncRNA in monocytes were determined via loss-of-function RNAi or gain-of-function lentivirus assays. The expression levels of mRNAs or proteins were evaluated using qRT-PCR, western blotting assay, or enzyme linked immunosorbent assays (ELISA). Results: A total of 1,042 mRNA transcripts (630 up-regulated and 412 down-regulated) were identified being differentially expressed between ASC and CHB patients. Through enrichment analysis we focused on the transforming growth factor beta (TGF-ß) signaling pathway and validated their expression in a larger cohort. Moreover, we found that lncRNA ENST00000519726 (lncRNA-HEIM) was highly expressed in monocytes and further up-regulated upon HBV infection. LncRNA-HEIM played an important role in CHB patients with long-term antiviral treatments, and its elevated expression was remarkably correlated with the TGF-ß signaling pathway, especially with the two members namely TGF-ß and SMAD4. Furthermore, altering the endogenous lncRNA-HEIM level in monocytes significantly affected the production of TGF-ß, as well as the fibrosis of hepatic stellate cells by affecting the expression of collagen I and α-smooth muscle actin (α-SMA). Conclusion: These findings not only added knowledge to the understanding of the roles of which lncRNA-HEIM played in the activation of HSCs in CHB patients with long-term medication, but also provided a promising therapeutic target in the future treatment for liver fibrosis.

Correction to: Human ex vivo spinal cord slice culture as a useful model of neural development, lesion, and allogeneic neural cell therapy.

An amendment to this paper has been published and can be accessed via the original article.

Human ex vivo spinal cord slice culture as a useful model of neural development, lesion, and allogeneic neural cell therapy.

BACKGROUND: There are multiple promising treatment strategies for central nervous system trauma and disease. However, to develop clinically potent and safe treatments, models of human-specific conditions are needed to complement in vitro and in vivo animal model-based studies. METHODS: We established human brain stem and spinal cord (cross- and longitudinal sections) organotypic cultures (hOCs) from first trimester tissues after informed consent by donor and ethical approval by the Regional Human Ethics Committee, Stockholm (lately referred to as Swedish Ethical Review Authority), and The National Board of Health and Welfare, Sweden. We evaluated the stability of hOCs with a semi-quantitative hOC score, immunohistochemistry, flow cytometry, Ca2+ signaling, and electrophysiological analysis. We also applied experimental allogeneic human neural cell therapy after injury in the ex vivo spinal cord slices. RESULTS: The spinal cord hOCs presented relatively stable features during 7-21 days in vitro (DIV) (except a slightly increased cell proliferation and activated glial response). After contusion injury performed at 7 DIV, a significant reduction of the hOC score, increase of the activated caspase-3+ cell population, and activated microglial populations at 14 days postinjury compared to sham controls were observed. Such elevation in the activated caspase-3+ population and activated microglial population was not observed after allogeneic human neural cell therapy. CONCLUSIONS: We conclude that human spinal cord slice cultures have potential for future structural and functional studies of human spinal cord development, injury, and treatment strategies.

Clinical characteristics and factors influencing the prognosis of Behçet's disease complicated with vascular involvement.

Background: Vascular Behçet's disease (VBD) might involve all sizes of arterial and venous vessels. Major vascular involvement caused the primary death in Behçet's syndrome (BS). We aimed to investigate the clinical characteristics and factors influencing the prognosis of VBD. Patients and methods: A retrospective analysis of the prospectively collected data of the Shanghai BS database from October 2012 to October 2018 was conducted. Patients who were diagnosed with BS and merged with venous thrombosis, arterial aneurysms, and arterial stenosis/occlusions were enrolled. Results: There were 47 patients with vascular involvement among 836 BS patients, 38 males and 9 females. The numbers of patients with venous thrombosis, arterial aneurysm, and arterial stenosis/occlusion were 25 (53.2 %), 21 (44.7 %), and 12 (25.5 %), respectively. Nearly half of the venous thromboses were located in limbs (n = 22, 46.8 %). Arterial aneurysm was the main form of arterial lesion. Most of the patients (93.6 %) were treated with corticosteroids and immunosuppressants. Late onset of BS or with arterial involvement had lower treatment response. Therapy with biological agents had significantly better results than that in the group without biological treatment (94.1 % vs. 80 %, P = 0.005). Conclusions: VBD showed a male preponderance and more than half of the patients presented with venous thrombosis. Late onset and arterial involvement were associated with poor prognosis. Therapy with biological agents is a viable alternative treatment to improve the prognosis.

Poor prognostic factors in patients with newly diagnosed intestinal Adamantiades-Behçet's disease in the Shanghai Adamantiades-Behçet's disease database: a prospective cohort study.

BACKGROUND: Adamantiades-Behçet's Disease (ABD) is an immunological recurrent systemic vasculitis with a chronic course. We investigated the predictors of long-term flare-ups, poor outcomes and event-free survival in Chinese non-surgical patients with intestinal ABD. METHODS: This was a prospective cohort study of 109 intestinal ABD patients seen in our institution between October 2012 and January 2019 who met the international criteria for ABD and had intestinal ulcers confirmed on colonoscopy. Predictors of relapses and poor outcomes, event-free survival were calculated using logistic regression models and Cox proportional hazard regression models, respectively. RESULTS: Sixty-six intestinal ABD patients (60.55%) had ileocecal ulcers; 19 patients (17.43%) presented with colorectum ulcers; 24 patients (22.02%) showed both ileocecal and colorectum ulcers. 7 patients (6.42%) experienced at least 1 flare-up of intestinal ulcers. 38 patients (34.86%) complained of non-healing intestinal ulcers. In multivariate analysis, location of intestinal ulcers (ileocecal and colorectum) (odd ratio (OR) 7.498 [95% confidence interval [95% CI] 1.844-30.480]), erythrocyte sedimentation rate (ESR) > 24 mm/h (OR 5.966 [95% CI 1.734-20.528]), treatment with infliximab (IFX) (OR 0.130 [95% CI 0.024-0.715]), and poor compliance (OR 11.730 [95% CI 2.341-58.781]) were independently correlated with a poor outcome. After a median follow-up of 28 months, 45 intestinal ABD patients (41.28%) underwent adverse events. Factors independently associated with shorter event-free survival were early onset of ABD (< 7 years) (hazard ratio (HR) 2.431 [95% CI 1.240-4.764]) and poor compliance (HR 3.058 [95% CI 1.612-5.800]). CONCLUSION: Distribution of intestinal ulcers (ileocecal and colorectum), ESR > 24 mm/h, treatment without IFX, and poor compliance were independent risk factors for poor outcomes in non-surgical intestinal ABD patients.

High expression of APC is an unfavorable prognostic biomarker in T4 gastric cancer patients.

BACKGROUND: Adenoma polyposis coli (APC) mutation is associated with tumorigenesis via the Wnt signaling pathway. AIM: To investigate the clinical features and mechanism of APC expression in gastric cancer (GC). METHODS: Based on APC expression profile, the related genome-wide mRNA expression, microRNA (miRNA) expression, and methylation profile in GC, the relationship between APC and GC, as well as the prognostic significance of APC were systematically analyzed by multi-dimensional methods. RESULTS: We found that high expression of APC (APC high) was significantly associated with adverse outcomes of T4 GC patients. Genome-wide gene expression analysis revealed that varying APC expression levels in GC were associated with some important oncogenes, and corresponding cellular functional pathways. Genome-wide miRNA expression analysis indicated that most of miRNAs associated with high APC expression were downregulated. The mRNA-miRNA regulatory network analysis revealed that down-regulated miRNAs affected their inhibitory effect on tumor genes. Genome-wide methylation profiles associated with APC expression showed that there was differential methylation between the APC high and APC low groups. The number of hypermethylation sites was larger than that of hypomethylation sites, and most of hypermethylation sites were enriched in CpG islands. CONCLUSION: Our research demonstrated that high APC expression is an unfavorable prognostic factor for T4 GC patients and may be used as a novel biomarker for pathogenesis research, diagnosis, and treatment of GC.

Evidence-based analysis on the toxicity of disinfection byproducts in vivo and in vitro for disinfection selection.

Disinfection is a key step in drinking water treatment process to prevent water-borne infections. However, reactions between chlorine, one of the most common disinfectants, and natural organic matter (NOM) often lead to the formation of hazardous disinfection byproducts (DBPs). However, the cytotoxicity of some DBPs is still poorly understood. Such knowledge is critical for proper selection of disinfection processes. We investigated the effects of DBPs on mouse acute liver injury. The exacerbation of liver damage increased with the DBPs concentrations, likely due to the increased hepatic macrophages. Haloacetonitriles (HANs) and haloketones (HKs) are more toxic to Human Hepatocellular (Hep3B) cells than trihalomethanes (THMs). Cytotoxicity of DBPs were governed by the halogen type (brominated DBPs > chlorinated DBPs) and the numbers of halogen atoms per molecule. Then, we used the pilot-scale WDS to study the best conditions for reducing the formation of DBPs. The result showed that the formation of DBPs followed the order: stainless-steel (SS) > ductile iron (DI) > polyethylene (PE) pipe. Higher flowrate promoted the formation of DBPs in all three pipes. The results suggest that the formation of DBPs in chlorine disinfection can be reduced by using PE pipes and low flow rate in water distribution systems (WDS).