Riboflavin Attenuates Fluoride-Induced Testicular Injury via Interleukin 17A-Mediated Classical Pyroptosis.

Male reproductive toxicity of fluoride is of great concern worldwide, yet the underlying mechanism is unclear. Pyroptosis is a novel mode of inflammatory cell death, and riboflavin with anti-inflammatory properties has the potential to protect against fluoride damage. However, it is unknown whether pyroptosis is involved in fluoride-induced testicular injury and riboflavin intervention. Here, we first found that riboflavin could alleviate fluoride-caused lower sperm quality and damaged testicular morphology by reducing pyroptosis based on a model of ICR mice treated with NaF (100 mg/L) and/or riboflavin supplementation (40 mg/L) via drinking water for 13 weeks. And then, together with the results of in vitro Leydig cell modelsm it was confirmed that the pyroptosis occurs predominantly through classical NLRP3/Caspase-1/GSDMD pathway. Furthermore, our results reveal that interleukin-17A mediates the process of pyroptosis in testes induced by fluoride and riboflavin attenuation according to the results of our established models of riboflavin- and/or fluoride-treated IL-17A knockout mice. The results not only declare a new mechanism by which fluoride induces testicular injury via interleukin 17A-mediated classical pyroptosis but also provide evidence for the potential clinical application of riboflavin as an effective therapy for fluoride toxicity.

Transfer learning with data alignment and optimal transport for EEG based motor imagery classification.

Objective. The non-stationarity of electroencephalogram (EEG) signals and the variability among different subjects present significant challenges in current Brain-Computer Interfaces (BCI) research, which requires a time-consuming specific calibration procedure to address. Transfer Learning (TL) offers a potential solution by leveraging data or models from one or more source domains to facilitate learning in the target domain, so as to address these challenges.Approach. In this paper, a novel Multi-source domain Transfer Learning Fusion (MTLF) framework is proposed to address the calibration problem. Firstly, the method transforms the source domain data with the resting state segment data, in order to decrease the differences between the source domain and the target domain. Subsequently, feature extraction is performed using common spatial pattern. Finally, an improved TL classifier is employed to classify the target samples. Notably, this method does not require the label information of target domain samples, while concurrently reducing the calibration workload.Main results. The proposed MTLF is assessed on Datasets 2a and 2b from the BCI Competition IV. Compared with other algorithms, our method performed relatively the best and achieved mean classification accuracy of 73.69% and 70.83% on Datasets 2a and 2b respectively.Significance.Experimental results demonstrate that the MTLF framework effectively reduces the discrepancy between the source and target domains and acquires better classification performance on two motor imagery datasets.

Melatonin alleviated fluoride-induced impairment of spermatogenesis and sperm maturation process via Interleukin-17A.

Fluoride-induced male reproductive failure is a major environmental and human health concern, but interventions are still lacking. Melatonin (MLT) has potential functions in regulating testicular damage and interleukin-17 (IL-17) production. This study aims to explore whether MLT can mitigate fluoride-induced male reproductive toxicity through IL-17A, and screen the potential targets. So the wild type and IL-17A knockout mice were employed and treated with sodium fluoride (100 mg/L) by drinking water and MLT (10 mg/kg.BW, intraperitoneal injection per two days starting from week 16) for 18 weeks. Bone F- concentrations, grade of dental damage, sperm quality, spermatogenic cells counts, histological morphology of testis and epididymis, and the mRNA expression of spermatogenesis and maturation, classical pyroptosis related and immune factor genes were detected respectively. The results revealed that MLT supplementations alleviated fluoride-induced impairment of spermatogenesis and maturation process, protecting the morphology of testis and epididymis through IL-17A pathway, and Tesk1 and Pten were identified as candidate targets from 29 regulation genes. Taken together, this study demonstrated a new physiological role for MLT in the protection against fluoride-induced reproductive injury and possible regulation mechanisms, which providing a useful therapeutic strategy for male reproductive function failure caused by fluoride or other environmental pollutants.

Potential Protective Effect of Riboflavin Against Pathological Changes in the Main Organs of Male Mice Induced by Fluoride Exposure.

Long-term exposure to excessive fluorine could cause damage to various tissues and organs in human and animals. However, there is no effective antidote to prevent and cure fluorosis except for avoiding fluoride intake. As an essential nutrient, riboflavin (VB2) has been identified to relieve oxidative stress and inflammation in animal tissues caused by other toxic substances, whether it can alleviate the damage caused by fluoride is unknown. For this, 32 ICR male mice were allocated to four groups of eight each. They were treated with 0 (distilled water), 100 mg/L sodium fluoride (NaF), 40 mg/L VB2, and their combination (100 mg/L NaF plus 40 mg/L VB2) via the drinking water for 90 consecutive days, respectively. The content of bone fluoride and the histomorphology of the main organs including liver, kidney, cerebral cortex, epididymis, small intestine, and colon were evaluated and pathologically scored. The results found that fluoride caused the pathological changes in liver, kidney, cerebral cortex, epididymis, small intestine, and colon at varying degrees, while riboflavin supplementation reduced significantly the accumulation of fluoride in bone, alleviated the morphological damage to cerebral cortex, epididymis, ileum, and colon. This study provides new clues for deeply exploring the mechanism of riboflavin intervention in fluorosis.

[Transport characteristics of Shuxiong prescription across Caco-2 cell monolayer model].

Shuxiong prescription (Notoginseng Radix et Rhizoma, Chuanxiong Rhizome and Carthami Flos) has the function of activating blood circulation to dissipate blood stasis, activating meridians to stop pain. This paper was mainly aimed to discuss the transport characteristics of Shuxiong prescription across Caco-2 cell monolayer. Safe concentration range of Shuxiong prescription against Caco-2 cell monolayer model was determined by MTT assay. The mechanism of Shuxiong prescription bidirectional transport was investigated by Caco-2 cell monolayer model. The apparent permeability coefficient Papp of digoxin was determined by high performance liquid chromatography (HPLC). The test results showed that the Papp of extract from Notoginseng Radix et Rhizoma, Chuanxiong Rhizome, Carthami Flos, Chuanxiong Rhizome+Carthami Flos and Shuxiong prescription transport from apical (AP) side to basolateral (BL) side was (3.12±0.73)×10⁻6, (2.58±0.41)×10⁻6, (4.97±0.64)×10⁻6, (4.63±0.57)×10⁻6, (5.79±0.68)×10⁻6 cm·s⁻¹, respectively, indicating that the transport of digoxin across Caco-2 cell monolayer model was active absorption, and the P-gp protein took part in the process. Chuanxiong Rhizome could significantly decrease the transport of digoxin from BL→AP(P<0.01) and increase its transport from AP→BL(P<0.05) significantiy. After the addition of Shuxiong prescription, the transport of digoxin from BL→AP was significantly inhibited(P<0.01). The results suggested that the extract of safflower had no effect on P-gp transport, nor on the independence diffusion of digoxin. The transport of digoxin could be degraded by the extract of Chuanxiong Rhizome and the extract of Shuxiong prescription from BL→AP(P<0.01), significantly; pseudo-ginseng had no effect on the independence diffusion of digoxin; the extract of safflower+Chuanxiong Rhizome had the same experimental result as Chuanxiong Rhizome extract.

Analysis of the roles of dietary protein and calcium in fluoride-induced changes in T-lymphocyte subsets in rat.

The roles of dietary protein (Pr) and calcium (Ca) levels on the changes in T-lymphocyte subsets induced by excessive fluoride (F) intake were assessed using rats that were malnourished for 120 days as a model. The CD4+ and CD8+ T-lymphocytes in the spleen tissue were determined by flow cytometry and immunofluorescence assay. The percentages of CD3+ , CD4+ , and CD8+ T-lymphocytes were reduced in the spleen of rats exposed to excessive F, and malnutrition aggravated these changes in the T-lymphocytes. In addition, the mRNA expression levels of IL-1ß, IL-2, IL-6, TNF-α, and IFN-γ in the spleen were downregulated significantly. We also reported herein the increased apoptosis ratio following caspase-9 and caspase-3 upregulation in the spleen of rats exposed to excessive amount of F. Light and transmisison electron microscopy revealed the irregularly arranged lymphocytes, few lymph nodules and the apoptotic characteristic of lymphocytes, which are caused by the increased expression of caspase. In addition, Pr and Ca supplementation reversed the morphologic and T-lymphocytic changes in spleen under malnutrition. Taken together, our results revealed an endogenous caspase-mediated mechanism of regulating the apoptosis of the T-lymphocyte subsets, as well as the immune-related cytokine secretion, which reduces the immune function in F-induced rats. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1587-1595, 2017.

Choline supplementation alleviates fluoride-induced testicular toxicity by restoring the NGF and MEK expression in mice.

Fluoride is known to cause male reproductive toxicity, and the elucidation of its underlying mechanisms is an ongoing research focus in reproductive toxicology and epidemiology. Choline, an essential nutrient, has been extensively studied for its benefits in nervous system yet was rarely discussed for its prospective effect in male reproductive system. This study aims to explore the potential protective role of choline against NaF-induced male reproductive toxicity via MAPK pathway. The male mice were administrated by 150mg/L NaF in drinking water, 5.75g/kg choline in diet, and their combination respectively from maternal gestation to postnatal 15weeks. The results showed that fluoride exposure reduced body weight growth, lowered sperm count and survival percentages, altered testicular histology, down-regulated the mRNA expressions of NGF, Ras, Raf, and MEK genes in testes, as well as significantly decreased the expressions of both NGF and phosphor-MEK proteins in testes. Examination of data from choline-treated mice revealed that choline supplementation ameliorated these fluoride-induced changes. Taken together, our findings suggest that choline supplementation alleviates fluoride-induced testicular toxicity by restoring the NGF and phosphor-MEK expression. The suitable dosage and supplementation periods of choline await further exploration.

Comparison Analysis: Granger Causality and New Causality and Their Applications to Motor Imagery.

In this paper we first point out a fatal drawback that the widely used Granger causality (GC) needs to estimate the autoregressive model, which is equivalent to taking a series of backward recursive operations which are infeasible in many irreversible chemical reaction models. Thus, new causality (NC) proposed by Hu et al. (2011) is theoretically shown to be more sensitive to reveal true causality than GC. We then apply GC and NC to motor imagery (MI) which is an important mental process in cognitive neuroscience and psychology and has received growing attention for a long time. We study causality flow during MI using scalp electroencephalograms from nine subjects in Brain-computer interface competition IV held in 2008. We are interested in three regions: Cz (central area of the cerebral cortex), C3 (left area of the cerebral cortex), and C4 (right area of the cerebral cortex) which are considered to be optimal locations for recognizing MI states in the literature. Our results show that: 1) there is strong directional connectivity from Cz to C3/C4 during left- and right-hand MIs based on GC and NC; 2) during left-hand MI, there is directional connectivity from C4 to C3 based on GC and NC; 3) during right-hand MI, there is strong directional connectivity from C3 to C4 which is much clearly revealed by NC than by GC, i.e., NC largely improves the classification rate; and 4) NC is demonstrated to be much more sensitive to reveal causal influence between different brain regions than GC.

Regulatory Effect of Catalpol on Th1/Th2 cells in Mice with Bone Loss Induced by Estrogen Deficiency.

PROBLEM: Estradiol (E2 ) deficiency can cause bone loss and the skew of Th1/Th2 cells. However, the correlation between the Th1/Th2 cells and the bone loss induced by estrogen deficiency remains unclear. Our aim was to investigate the role of Th1/Th2 in bone loss induced by estrogen deficiency and elucidated the therapeutical effect of catalpol in this condition. METHOD OF STUDY: Young, sham-operated (Sham), ovariectomized (Ovx), and naturally aged mice, treated with catalpol at different doses or control vehicle, were used in this study as indicated in each experiment. ELISA assay, dual-energy X-ray absorptiometry, and flow cytometry were used to analyze E2 , C-terminal telopeptides of type I collagen (CTx-I), bone mineral density (BMD), and Th1/Th2 subsets, respectively. The mRNA and protein expressions of specific transcription factors for Th1/Th2 cells (T-bet and GATA-3) were analyzed using real-time quantitative PCR and Western blot, respectively. RESULTS: Bone mineral density and E2 levels positively correlated with the proportion of Th2 subset while negatively correlated with that of Th1 subset and the ratio of Th1/Th2. Catalpol alleviated bone loss effectively by regulating Th1/Th2 polarization. Catalpol promoted the expression of Th2-specific transcription factors while inhibited that associated with Th1. CONCLUSION: Th1/Th2 skew is involved in bone loss induced by estrogen deficiency. Catalpol alleviates bone loss effectively by regulating Th1/Th2 paradigm.

[Effect of different compatibility of tetramethylpyrazine on its pharmacokinetics in vivo].

OBJECTIVE: To study the pharmacokinetics of single administration and different compatibility of tetramethylpyrazine (TMP) in rats. METHOD: Thirty two Sprague-Dawley rats were randomly divided into 4 groups: the TMP (30 mg x kg(-1)) group, the TMP+FA (30 mg x kg(-1) + 50 mg x kg(-1)) group, the TMP+TET (30 mg x kg(-1) mg x kg(-1)) group and the TMP+FA+TET (30 mg x kg(-1) + 50 mg x kg(-1) + 20 mg x kg(-1)) group. After the oral administration, their blood samples were collected to detect plasmas concentrations by HPLC method and to calculate pharmacokinetic parameters DAS 2.0 program. RESULT: In compatibility with FA, AUC(0-t), Cmax and Tmax showed no significant difference with the single administration of TMP, but t(1/2) and MRT,, were obviously longer than the single administration. In compatibility with TET and FA + TET, AUC (0-t), Cmax and Tmax showed significant increase than the single administration of TMP, whereas t(1/2) and MRT0, did not notably vary from the single administration. CONCLUSION: FA can prolong TMP's action time in rats, and TET can accelerate TMP's absorption in rats.

[Comparative study on pharmacokinetics of tetramethylpyrazine, ferulic acid and their compatibility].

OBJECTIVE: To study the pharmacokinetics of tetramethylpyrazine (TMP), ferulic acid and their compatibility. METHOD: Twenty-four Sprague-Dawley rats were randomly divided into 3 groups: TMP 20 mg x kg(-1), ferulic acid 20 mg x kg(-1) and TMP 20 mg x kg(-1) + ferulic acid 20 mg x kg(-1). All the rats were given intragastric administration then blood samples were obtained from fossa orbitalis at several time points. All the plasmas concentrations were analyzed by HPLC method and the data were treated by DAS 2.0 program. RESULT: The main pharmacokinetics parameters of TMP 20 mg x kg(-1) group, ferulic acid 20 mg x kg(-1) and TMP 20 mg x kg(-1) + ferulic acid 20 mg x kg(-1) were as follows: t(max) 0.5 h, t1/2 0.856 h,MRT 1.321 h, AUC 5.112 microg x h(-1) x L(-1), C(max) 2.834 microg x L(-1); t(max) 0.083 h, t1/2 1.024 h, MRT 1.324 h, AUC 1.581 microg x h(-1) x L(-1), C(max) 1.492 microg x L(-1); t(max) 0.583 h, t1/2 37.901 h, MRT 3.798 h, AUC 4.097 microg x h(-1) x L(-1), C(max)1.571 microg x L(-1); t(max) 0.6 h, t1/2 7.860 h, MRT 2.894 h, AUC 1.984 microg x h(-1) x L(-1), C(max) 1.03 microg x L(-1), respectively. CONCLUSION: The experiments suggested that the compatibility of TMP and ferulic acid had interaction in pharmacokinetics; all the t1/2 and MRT were prolonged and had the effect of lente liberates.