Identification of core genes, critical signaling pathways, and potential drugs for countering BPA-induced hippocampal neurotoxicity in male mice.

Although the neurotoxicity of the common chemical bisphenol A (BPA) to the mouse hippocampus has been often reported, the mechanism underlying BPA-induced depression-like behavior in mice remains unclear. We evaluated BPA's role in inducing depressive-like behavior by exposing male mice to different BPA concentrations (0, 0.01, 0.1, and 1 µg/mL) and using the forced swimming test (FST) and tail suspension test (TST). We aimed to identify critical gene and anti-BPA-neurotoxicity compounds using RNA sequencing combined with bioinformatics analysis. Our results showed that 1 µg/mL BPA exposure increased mouse immobility during the FST and TST. Based on BPA-induced hippocampal transcriptome changes, we identified NADH: ubiquinone oxidoreductase subunit AB1 (Ndufab1) as a critical and potential therapeutic target gene, and Ndufab1 mRNA and protein levels were downregulated in the BPA-exposed groups. Furthermore, molecular docking identified phenelzine as a compound that could counteract BPA-related neurotoxicity. Conclusively, our analyses confirmed that BPA triggers depressive behavior in male mice by downregulating Ndufab1 expression and suggested that phenelzine might reduce BPA-induced neurotoxicity.

[Exploration of an online and offline mixed teaching mode for "Protein Engineering" based on BOPPPS+flipped classroom].

Protein Engineering is a core compulsory course of biotechnology major, which is the first-class undergraduate major being constructed in Shanxi Province. In view of the problems of single teaching mode of Protein Engineering, such as insufficient students' participation, short teaching time, and expensive experiment cost, the course team carried out the reform and practice of teaching mode for this course, and put forward a new teaching strategy. Under the guidance of the "Golden Course" standard for advancement, innovation and challenge, the course team developed the materials for massive open online courses (MOOC), and carried out the online and offline mixed teaching of Protein Engineering based on BOPPPS+flipped classroom by using the Chao-Xing Fan-Ya network teaching platform. Through this, a comprehensive, systematic and dynamic new teaching system of Protein Engineering was developed. Using the teaching mode based on BOPPPS+flipped classroom, the offline classroom teaching was combined with students' online self-study and homework completion, chapter test and discussion, and this mixed teaching mode was fully integrated into the flipped classroom. After three rounds of teaching practice, the course team had developed a complete, reproducible, scientific and reasonable online and offline mixed teaching mode, which included course materials preparation, exploring experiment guidance, classroom discussion design and course performance evaluation. The online and offline mixed teaching mode of Protein Engineering based on BOPPPS+flipped classroom was helpful for students to improve their autonomous learning ability, to be deeply engaged in the whole teaching process, and to develop a comprehensive and profound understanding of Protein Engineering. This teaching mode improved the teaching quality of Protein Engineering, and facilitated students to learn other follow-up professional courses. Moreover, it provides a reference for the course teaching reform.

[Mechanism of nephrotoxicity induced by chronic exposure of bisphenol A in mice based on oxidative stress and cell apoptosis].

Bisphenol A (BPA) is widely used to produce epoxy resin and polycarbonate plastic products. In severe cases, these plastics may release BPA, which then infiltrates into the environment. Various concentrations of BPA have been found in most biological fluid. Its presence has been well shown to be closely related to many chronic diseases, including chronic kidney disease (CKD). However, little is known regarding the adverse effects of BPA exposure and its succedent cellular events on CKD. Hence, in the current in vivo study, we aimed to assess the effects of chronic exposure to BPA on animal nephrotoxicity through investigating oxidative stress and apoptosis. Upon exposure to BPA at 0.01, 0.1, and 1 mg/L via drinking water for four weeks, the mated and pregnant females were continuously exposed to BPA until weaning. Subsequently, three weeks old F1-male neonates were also orally challenged with the same three doses of BPA for ten weeks. The results showed that the kidneys of 0.1 and 1 mg/L BPA-treated mice were seriously damaged; the contents of serum renal function indexes and lipid peroxidation products were significantly increased, including urea nitrogen, creatinine, uric acid, and thiobarbituric acid reactive substances; the morphological structure of mouse kidneys was impaired; the expressions of antioxidant-related genes at mRNA and protein levels from mouse kidneys were markedly diminished, including glutathione-S-transferase, superoxide dismutase, and catalase; the expressions of genes and proteins related to apoptosis index (ratio of Bax/Bcl-1 and Caspase-3) were significantly enhanced. The data manifested that cumulative oxidative stress and apoptosis might play an essential role in the animal nephrotoxicity induced by chronic exposure to BPA.

Bisphenol a exposure decreases learning ability through the suppression of mitochondrial oxidative phosphorylation in the hippocampus of male mice.

Bisphenol-A (BPA) is ubiquitous in the environment. Many studies have demonstrated that BPA triggers central nervous system dysfunction in animals, however, the molecular mechanisms of BPA neurotoxicity are unclear. Hence, this study aimed to evaluate the neurotoxic effects of male mouse hippocampus under BPA exposure (0 mg/L, control group (C); 0.01 mg/L, low dose (L); 0.1 mg/L, medium dose (M); and 1 mg/L, high dose (H)). Nine differentially expressed genes (DEG) related to oxidative phosphorylation (OXPHOs) were screened out and located through an interaction network and co-enrichment analysis. Overall, BPA exposure disrupted the regular cell arrangement in the hippocampus, reduced learning abilities and the ratio of testosterone (T)/estradiol (E2), inhibited gene expressions concerned OXPHOs, especially reduced protein expression level of the Ndufb10 related to learning ability and the activities of complex I, III, IV, and Ⅴ, which ultimately caused abnormal ATP synthesis of hippocampal mitochondria and led to shrinking rapidly myocardial energy supply in the brain (hippocampus). These results indicated that the destruction of mitochondrial OXPHOs might be one of the mechanisms of BPA-induced learning ability decline in mice. In conclusion, our results provide a creative understanding and research direction for the molecular alterations and signal pathways of BPA-induced neurotoxicity.

Chronic exposure of bisphenol-A impairs cognitive function and disrupts hippocampal insulin signaling pathway in male mice.

Bisphenol-A (BPA), a well-known estrogenic endocrine disruptor, is generally applied to turn out plastic consumer products. Available data have manifested that exposure to BPA can trigger insulin resistance. Hence, the purpose of the actual study was to consider the impacts of BPA exposure on cognitive function and insulin signaling pathway in the hippocampus of male offspring mice. For this purpose, the pregnant female mice were treated either vehicle (0.1% ethanol) or BPA (0.01, 0.1, and 1 µg/mL) via drinking water from day 1 of gestation until delactation (D1-PND21, newborn exposure). Afterward, the three-week-old male offspring mice took orally with the same doses of BPA for nine weeks (PND84). The behavioral tests, blood sugar level, histological observation, transcriptome sequencing, glucose transporter 4 (GLUT4), and hippocampal insulin signaling pathway were checked for the male offspring mice at 13 weeks of age (PND91). Our data indicated that BPA exposure impaired cognitive function, disrupted the hippocampal regular cell arrangement, increased blood glucose levels, disturbed the insulin signaling pathway including phosphorylated insulin receptor substrate1 (p-IRS1), protein kinase B (p-AKT), and glycogen synthase kinase 3ß (p-GSK3ß). At the same time, the mRNA and protein expressions of GLUT4 were markedly down-regulated in the BPA-exposed groups. To sum up, it has been suggested from these results that BPA has detrimental effects on the insulin signaling pathway, which might subsequently be conducive to the impairment of cognitive function in the adult male offspring mice. Therefore, BPA exposure might in part be an element of risk for the long-term neurodegeneration in male offspring mice.

Detrimental Effects of Sodium Fluoride on the Expression of Insulin Receptor in the Olfactory Bulb and Hippocampus of Male Mice.

Fluoride exposure is associated with lowered cognitive function ability, intelligence quotient, and mental decline, especially in children. The brain insulin receptor (IR) signaling system is related to neuronal plasticity and consequent cognitive ability. In our previous study, NaF exposure decreased IR expressions in olfactory bulb (OB) and hippocampus after Y-maze test in male mice. In order to further explore whether the Y-maze test affected IR gene and protein expression levels in the OB and hippocampus under the NaF exposure, healthy male mice were randomly allotted into four groups and challenged with 0, 50, 100, and 150 mg/L NaF for three continuous months. The results showed that femur fluorine content of the NaF-exposed groups increased significantly in a dose-dependent manner. NaF significantly decreased brain protein content and organ coefficient of the treated male mice. The protein and mRNA expression levels of the IR were significantly decreased in the OB and hippocampus of the NaF-treated mice. Interestingly, indicators (brain protein content and organ coefficient) measured in the present study were significantly lower than our previous study indicators (mice tested Y-maze test), especially the expression levels of IR protein and mRNA in the same concentration groups. Taken together, these results indicated that Y-maze test could promote the expression levels of IR protein and mRNA in the OB and hippocampus, while NaF had a stronger inhibitory effect, which resulted in adverse effects on the expression levels of IR in the OB and hippocampus of male mice.

Bisphenol A-induced apoptosis, oxidative stress and DNA damage in cultured rhesus monkey embryo renal epithelial Marc-145 cells.

Bisphenol A (BPA) is widely used in the production of epoxy resins and polycarbonate plastics. Under harsh situations, these plastics likely desorb BPA, which then can seep into the environment. Various concentrations of BPA have been detected in most biological fluid. However, there is paucity of information on the detrimental effects of BPA and its subsequent cellular events in chronic kidney disease (CKD). Hence, in this in vitro study, we aimed to investigate the effects of BPA on renal epithelial cell activation, apoptosis, and DNA damage. Rhesus monkey embryo renal epithelial Marc-145 cells were exposed to 0, 10-1, 10-2, 10-3, 10-4, 10-5, and 10-6 M of BPA. Alterations in intracellular apoptosis, oxidative stress, and DNA damage were evaluated. The results showed that BPA decreased cell viability, superoxide dismutase (SOD) activity and glutathione (GSH) level, with concomitant increases in apoptosis related indices, lactate dehydrogenase (LDH) activity, reactive oxygen species (ROS) generation, thiobarbituric acid reactive substances (TBARS) content, and the rate of comet Marc-145 cells with a dose-dependent manner. The data indicated that increased oxidative stress, apoptosis and DNA damage in epithelial Marc-145 cells might play a pivotal role in the mechanism of BPA-induced nephrotoxicity.

Fluoride exposure decreased learning ability and the expressions of the insulin receptor in male mouse hippocampus and olfactory bulb.

Fluoride is one of the common environmental pollutants. Internal exposure to fluoride is related to the lowered cognitive function and intelligence, particularly for children. Determination of protein content in brain tissue is a means to reflect the functional development of the central nervous system. Insulin and insulin receptor (IR) signaling systems are associated with cognitive ability. The present research focused on the assessment of the expressions of IR protein and mRNA in hippocampus and olfactory bulb (OB), as well as learning and memory ability of male Kunming mice. Mice were exposed to 50, 100, and 150 mg/L NaF for 90 continuous days. The results showed that learning and memory abilities as well as protein content of male mice brain was significantly decreased by fluoride. Fluoride could inhibit the protein and mRNA expressions of the IR in the hippocampus and OB of mice. IRs mainly distributed in the olfactory nerve layer of the outermost layer of the OB, and most distributed in the hippocampal cornu ammon 3 (CA3) region, followed by the dentate gyrus (DG) and cornu ammon 1 (CA1) regions. These findings suggested that inhibition of the IR protein and mRNA expressions in the hippocampus and OB by fluoride might in part affect learning and memory ability in male mice.

[Analysis of exogenous gene and protein digestion and absorption of SD rats (Rattus norvegicus) fed roundup ready soybean meal].

Metabolism and deposition of exogenous gene and protein from transgenic glyphosate herbicide-tolerant soybean meal in SD rats were studied in the experiment. The transgenic soybean GTS40-3-2 meal and its non-transgenic counterpart (parent A5403) were fed to the generation and the second generation Sprague-Dawley (SD) rats (Rattus norvegicus). The study added the genetically modified (GM) soybean meal and its non-transgenic control soybean meal (parent A5403) in a ratio of 20% respectively to the feeds. By using qualitative, quantitative PCR and ELISA methods to detect transgenic soybean residues of metabolism ingredients in rats, the safety and influence of GM soybean were evaluated. The results showed that the intestinal fecal and cecum contents of rats were detected with residues of GM ingredients, intestinal flora and organs were not found related genes and protein. These results indicated that transgenic glyphosate herbicide-tolerant soybean GTS40-3-2 meal was as safe as its non-GM soybean meal in long-term feeding study.

[Detection of transgenic components in animal feeds on Shanxi markets].

To assess the presence of genetically modified (GM) maize and soybean in a range of commercialized feed in Shanxi province of China in 2015, improved hexadecyltrimethy ammonium bromide (CTAB) method was used to extract DNA. The screening of packed feeds was carried out by qualitative PCR. Then positive feeds were unpacked and detected by the CaMV 35S promoter, NOS terminator, zSSIIb, Lectin and CryIA (b) genes. The identified maize and soybean events were confirmed by event-specific MON810 and GTS40-3-2. Results showed that 83.3% of the feeds was tested positive for GMOs, in which positive rates of maize, soybean, pig and layer feeds were 6.67%, 100%, 93.3% and 73.3%, respectively. The results of real-time PCR were consistent with qualitative PCR. These results indicated that commercialized GM feed had a wide positive product scope in Shanxi province of China. Further studies are necessary to study effects of feeding livestock and poultry with feed containing GM ingredients on animals and their products.

Effects of low-level laser irradiation on rat skeletal muscle injury after eccentric exercise.

BACKGROUND AND OBJECTIVE: The effect of photobiomodulation on delayed onset muscle soreness remains unknown. This study represents the first investigation of this treatment using an animal model. METHODS: Seventy-two Sprague-Dawley rats were randomly divided into five groups: sedentary control group, exercise control group and three exercise-plus-laser groups. Downhill running was used to induce muscle injury in the gastrocnemius muscle. He-Ne laser irradiations were administered to the injured muscles immediately and at 18 and 42 h after exercise in the three exercise-plus-laser groups at 12, 28, and 43 J/cm2, respectively. Histological examination and serum creatine kinase (CK), muscle superoxide dismutase (SOD) and malondialdehyde (MDA) analyses were done at 24 and 48 h after exercise. RESULTS: The exercise control group exhibited a marked inflammation in the gastrocnemius muscle and significant elevations in serum CK activity and muscle MDA level after downhill running. He-Ne laser irradiation at 43 J/cm2 inhibited muscle inflammation, significantly enhanced muscle SOD activity and significantly reduced serum CK activity and muscle MDA level at both 24 and 48 h after exercise, whereas the irradiation at 12 or 28 J/cm2 slightly inhibited muscle inflammation and significantly reduced serum CK activity at 48 h after exercise only (P<0.05). CONCLUSIONS: Low-level He-Ne laser therapy could exert therapeutic effects on eccentric exercise-induced rat muscle injury through enhancing muscle anti-oxidative capacity and reducing the inflammatory reaction. The photobiomodulation was dose-dependent, and the 43 J/cm2 dose was the most efficient among the doses used.