Inhibition of urinary bladder cancer cell proliferation by silibinin

Silibinin, a natural compound extracted from milk thistle, has demonstrated antitumor properties in urinary bladder cancer cells; however, the role of TP53 gene in these effects is unclear. In order to better understand the molecular and antiproliferative mechanisms of this compound, urinary bladder cancer cells with different TP53 gene status, RT4 (low-grade tumor, wild TP53 gene), 5637 (high-grade tumor, Grade 2, mutated TP53 gene), and T24 (high-grade tumor, Grade 3, mutated TP53 gene) were treated with several concentrations of silibinin (1, 5, 10, 50, 100, and 150 μM). Cytotoxicity, prooxidant effect, morphological changes, cell migration, cell cycle progression, global methylation profile, and relative expression of HOXB3, c-MYC, PLK1, SMAD4, SRC, HAT, HDAC, and RASSF1A genes were evaluated. The silibinin presented cytotoxic and prooxidant effects in the three cell lines. In mutated TP53 cells, significant interference in cell migration and cell cycle arrest at the G2/M phase was observed. Additionally, silibinin induced global DNA hypomethylation in the highest grade tumor cells. For wild-type TP53 cells, a sub-G1 apoptotic population was present. Furthermore, there was modulation of gene expression responsible for cell growth (SMAD and c-MYC), migration (SRC), cell cycle kinetics (PLK1), angiogenesis (HOXB3), and of genes associated with epigenetic events such as DNA acetylation (HAT) and deacetylation (HDAC). In conclusion, the silibinin inhibited the urinary bladder tumor cell proliferation independently of TP53 status; however, cell cycle effects, gene expression changes, and alteration of cell migration are dependent on TP53 status. © 2020 Wiley Periodicals, Inc.     

膝关节自体软骨细胞移植术后康复的研究进展

目的总结膝关节自体软骨细胞移植术（autologous chondrocyte implantation，ACI）术后康复的研究进展。方法查阅近年来国内外 ACI 术后康复相关的基础及临床研究，并进行总结分析。结果研究表明，ACI 术后移植物成熟分为 4 个阶段：增生期（术后 0～6 周）、过渡期（术后 6～12 周）、重塑期（术后 12～26 周）、成熟期（术后 26 周～2 年）。为了避免早期移植物损伤，适当刺激移植物和骨骼的整合过程，更快恢复关节功能，临床需针对术后不同阶段移植物的生物力学特性设计康复锻炼方案，主要体现在术后负重、运动范围、康复训练方式的过渡和调整。结论膝关节 ACI 术后应科学地逐步增加负重和运动范围，在不同阶段可借助相应的康复锻炼手段。但目前相关研究的证据等级较低，仍需更高质量、更大样本量的随机对照试验，进一步探讨完善康复方案。     

Transurethral instillation with fusion protein MrpH.FimH induces protective innate immune responses against uropathogenic Escherichia coli and Proteus mirabilis

Urinary tract infections (UTIs) are among the most common infections in human. Innate immunity recognizes pathogen‐associated molecular patterns (PAMPs) by Toll‐like receptors (TLRs) to activate responses against pathogens. Recently, we demonstrated that MrpH.FimH fusion protein consisting of MrpH from Proteus mirabilis and FimH from Uropathogenic Escherichia coli (UPEC) results in the higher immunogenicity and protection, as compared with FimH and MrpH alone. In this study, we evaluated the innate immunity and adjuvant properties induced by fusion MrpH.FimH through in vitro and in vivo methods. FimH and MrpH.FimH were able to induce significantly higher IL‐8 and IL‐6 responses than untreated or MrpH alone in cell lines tested. The neutrophil count was significantly higher in the fusion group than other groups. After 6 h, IL‐8 and IL‐6 production reached a peak, with a significant decline at 24 h post‐instillation in both bladder and kidney tissues. Mice instilled with the fusion and challenged with UPEC or P. mirabilis showed a significant decrease in the number of bacteria in bladder and kidney compared to control mice. The results of these studies demonstrate that the use of recombinant fusion protein encoding TLR‐4 ligand represents an effective vaccination strategy that does not require the use of a commercial adjuvant. Furthermore, MrpH.FimH was presented as a promising vaccine candidate against UTIs caused by UPEC and P. mirabilis.     

CYTED Network to develop an immunogen compatible with integrated management strategies for tick control in cattle

INCOGARR is a thematic network recently approved to be financially supported by the Ibero-American Program of Science and Technology for Development (CYTED). The objectives of this Network are the design and evaluation of an efficient and feasible anti-tick vaccine candidate from the technical and economical points of view and also sharing experiences in the immunological control of ticks as part of an Integrated Control Program. The Network consists of seven laboratories and one company from six countries. The first meeting of the Network took place with the representation of each laboratory involved. In the meeting, general and specific objectives and activities of the Network were discussed and it was a very nice example of international collaboration to address an unsolved worldwide topic on tick control in which laboratories with different competencies and expertise join their efforts in a common goal.     

The time course of psychological stress as revealed by event-related potentials

Psychological stress is common in everyday life and is believed to affect emotion, cognition and health. Previous brain imaging studies have been able to identify the brain regions involved in the stress response. However, our understanding of the temporal neurological response to psychological stress is limited. The present work aims to investigate the time course of psychological stress induced by a mental arithmetic task, utilizing event-related potentials (ERPs). The elicitation of stress was verified by self-reports of stress and increases in salivary cortisol levels. The subjective and physiological data showed that the stress-elicitation paradigm successfully induced a mild-to-moderate level of psychological stress. The electrophysiological data showed that the amplitude of occipital N1 was more negative in the control task than in the stress task, and the latency of frontal P2 was shorter in the stress task than in the control task. Our results provide electrophysiological evidence that psychological stress occurs primarily at the early stage of cognitive processing.     

Is Subjective Outcome Evaluation Related to Objective Outcome Evaluation? Insights from a Longitudinal Study in Hong Kong

Study ObjectivesEmpirical studies investigating the inter-relationships among subjective outcomes and objective outcomes are sparse in the scientific literature. This study examined the relationship between these 2 forms of outcome evaluation data in Chinese adolescents, utilizing longitudinal data.DesignIn Wave 3 and Wave 4 of a randomized group trial, 2,784 Chinese secondary school students responded to the Chinese Positive Youth Development Scale (CPYDS) assessing objective outcome at pretest (Wave 3) and posttest (Wave 4). Students in the experimental schools also responded to a measure of client satisfaction assessing the subjective outcome of a positive youth development program at posttest (Wave 4).ResultsSubjective outcome evaluation measures (eg, process, program, and effectiveness) at Wave 3 were significantly correlated with CPYDS scores at Wave 4 and difference scores. Subjective outcome evaluation measures also predicted changes in the objective outcome evaluation measures scores across time.ConclusionsThe present findings replicated the previous findings that subjective outcome and objective outcome evaluation findings were intimately related, and subjective outcome measures predicted posttest measures and their changes over time. The present study suggests that subjective outcome evaluation findings may yield useful information regarding program effectiveness in youth work settings.     

Localization of TRPV1 and P2X3 in unmyelinated and myelinated vagal afferents in the rat

The vagus nerve is dominated by afferent fibers that convey sensory information from the viscera to the brain. Most vagal afferents are unmyelinated, slow-conducting C-fibers, while a smaller portion are myelinated, fast-conducting A-fibers. Vagal afferents terminate in the nucleus tractus solitarius (NTS) in the dorsal brainstem and regulate autonomic and respiratory reflexes, as well as ascending pathways throughout the brain. Vagal afferents form glutamatergic excitatory synapses with postsynaptic NTS neurons that are modulated by a variety of channels. The organization of vagal afferents with regard to fiber type and channels is not well understood. In the present study, we used tract tracing methods to identify distinct populations of vagal afferents to determine if key channels are selectively localized to specific groups of afferent fibers. Vagal afferents were labeled with isolectin B4 (IB4) or cholera toxin B (CTb) to detect unmyelinated and myelinated afferents, respectively. We find that TRPV1 channels are preferentially found in unmyelinated vagal afferents identified with IB4, with almost half of all IB4 fibers showing co-localization with TRPV1. These results agree with prior electrophysiological findings. In contrast, we found that the ATP-sensitive channel P2X3 is found in a subset of both myelinated and unmyelinated vagal afferent fibers. Specifically, 18% of IB4 and 23% of CTb afferents contained P2X3. The majority of CTb-ir vagal afferents contained neither channel. Since neither channel was found in all vagal afferents, there are likely further degrees of heterogeneity in the modulation of vagal afferent sensory input to the NTS beyond fiber type.     

Hepatoprotective and antioxidant potential of Asphodeline lutea (L.) Rchb. roots extract in experimental models in vitro/in vivo

The aim of this study was to investigate the effect of Asphodeline lutea (L.) Rchb. dry root extract (ALE) administered alone and against carbon tetrachloride (CCl₄)-induced liver injury in vitro/in vivo. The dried roots of A. lutea were extracted with 70% ethanol and was characterized with HPLC-UV. Hepatoprotective potential was investigated by in vivo/in vitro assays in Wistar rats as well as antioxidant properties. At concentrations ranging from 10 to 200 μg/mL of ALE significant cytotoxic effects on isolated hepatocytes were found. ALE showed some toxicity in Wistar rats discerned by increased ALT (Alanine transaminase), ALP (Alkaline phosphatase) activities and MDA (malondialdehyde) quantity, decreased GSH (reduced glutathione) levels without affecting the activity of the antioxidant enzymes (GPx (Gluthatione peroxidase), GR (Glutathione reductase) and GST (Glutathione-S-transferase activity)). The antioxidant and hepatoprotective potential of ALE was also observed in vitro/in vivo against CCl₄-induced liver injury, where ALE normalizes all the examined parameters perturbated by CCl₄ administration. In addition, ALE preserved the decreased cytochrome P450 level and EMND (Ethylmorphine-N-Demethylase) activity without affecting AH (Aniline 4-Hydroxylase) activity. ALE is rich in anthraquinones, naphthalenes and caffeic acid. The pro-oxidant effects of ALE could be due to naphthalene and anthraquinone bioactivation pathways involving toxic metabolites.