Pinoresinol diglucoside mitigates dexamethasone-induced osteoporosis and chondrodysplasia in zebrafish.

BACKGROUND: The global increase in the aging population has led to a higher incidence of osteoporosis among the elderly. OBJECTIVE: This study aimed to evaluate the protective properties of pinoresinol diglucoside (PDG), an active constituent of Eucommia ulmoides, against dexamethasone-induced osteoporosis and chondrodysplasia. METHODS: A zebrafish model of osteoporosis was established by exposing larval zebrafish to dexamethasone. The impact of PDG on bone mineralization was assessed through alizarin red and calcein staining. Alkaline phosphatase activity was quantified to evaluate osteoblast function. The influence of PDG on chondrogenesis was estimated using alcian blue staining. Fluorescence imaging and motor behavior analysis were employed to assess the protective effect of PDG on the structure and function of dexamethasone-induced skeletal teratogenesis. qPCR determined the expression of osteogenesis and Wnt signaling-related genes. Molecular docking was used to assess the potential interactions between PDG and Wnt receptors. RESULTS: PDG significantly increased bone mineralization and corrected spinal curvature and cartilage malformations in the zebrafish model. Furthermore, PDG enhanced swimming abilities compared to the model group. PDG mitigated dexamethasone-induced skeletal abnormalities in zebrafish by upregulating Wnt signaling, showing potential interaction with Wnt receptors FZD2 and FZD5. CONCLUSION: PDG mitigates dexamethasone-induced osteoporosis and chondrodysplasia by promoting bone formation and activating Wnt signaling.

Rapid Determination of Tetracyclines in Drinking and Environmental Waters Using Fully Automatic Solid-Phase Extraction with Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry.

The abuse and irrational use of tetracyclines (TCs) in human medicine and animal husbandry has become a serious concern, affecting the ecological environment and human health. The aim of this study was to develop a sensitive and selective method using fully automatic solid-phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry for the determination of twelve TCs in water. Four isotope-labeled internal standards for TCs were used to correct matrix effects. Several parameters affecting extraction efficiency were systematically optimized, and the optimum experimental conditions found were 1.0 L water sample with 0.5 g/L Na2EDTA (pH 3.0) extracted and enriched by CNW HLB cartridge and eluted by 4 mL of acetone:methanol (v/v, 1:1). The enrichment factors were up to 798-1059 but only requiring about 60 min per six samples. Under the optimized conditions, the linearity of the method ranged from 0.2 to 100 µg/L for 12 TCs, the detection limits were as low as 0.01-0.15 ng/L, and the recoveries were in the range of 70%-118%, with relative standard deviations less than 15%. The developed method can be successfully utilized for the determination of 12 TCs in pure water, tap water, river water, and mariculture seawater. In summary, three and six TCs were detected in river water and mariculture seawater, respectively, with total concentrations of 0.074-0.520 ng/L (mean 0.248 ng/L) and 0.792-58.369 ng/L (12.629 ng/L), respectively. Tetracycline (TC) and oxytetracycline (OTC) were the dominant TCs in river water, while doxytetracycline (DXC) and OTC were dominant in mariculture seawater.

Genetic variations in ZmSAUR15 contribute to the formation of immature embryo-derived embryonic calluses in maize.

The ability of immature maize (Zea mays) embryos to form embryonic calluses (ECs) is highly genotype dependent, which limits transgenic breeding development in maize. Here, we report the association map-based cloning of ZmSAUR15 using an association panel (AP) consisting of 309 inbred lines with diverse formation abilities for ECs. We demonstrated that ZmSAUR15, which encodes a small auxin-upregulated RNA, acts as a negative effector in maize EC induction. Polymorphisms in the ZmSAUR15 promoter that influence the expression of ZmSAUR15 transcripts modulate the EC induction capacity in maize. ZmSAUR15 is involved in indole-3-acetic acid biosynthesis and cell division in immature embryo-derived callus. The ability of immature embryos to induce EC formation can be improved by the knockout of ZmSAUR15, which consequently increases the callus regeneration efficiency. Our study provides new insights into overcoming the genotypic limitations associated with EC formation and improving genetic transformation in maize.

Cyhalofop-butyl exposure induces the severe hepatotoxicity and immunotoxicity in zebrafish embryos.

Cyhalofop-butyl (CyB) is a highly effective herbicide and is widely used for weed control in paddy fields. Because CyB is easily residual in the aquatic environment, its potential harm to aquatic organisms has attracted much attention and has not been fully understood. In this study, we systematically explored the hepatotoxic and immunotoxic effects of CyB exposure in zebrafish embryos. Firstly, CyB induced a decrease in the survival rate of zebrafish and led to a series of developmental abnormalities. Meanwhile, CyB can significantly reduce the size of zebrafish liver tissue and the number of hepatocytes in a dose-dependent manner. Secondly, the number of macrophages and neutrophils significantly decreased but the antioxidant enzyme activities such as CAT and MDA were greatly elevated upon CyB exposure. Thirdly, RNA-Seq analysis identified 1, 402 differentially expressed genes (DEGs) including 621 up-regulated and 781 down-regulated in zebrafish embryos after CyB exposure. KEGG and GO functional analysis revealed that the metabolic pathways of drug metabolism-cytochrome P450, biosynthesis of antibiotics, and metabolism of xenobiotics, along with oxidation-reduction process, high-density lipoprotein particle and cholesterol transport activity were significantly enriched after CyB exposure. Besides, hierarchical clustering analysis suggested that the genes involved in lipid metabolism, oxidative stress and innate immunity were largely activated in CyB-exposed zebrafish. Moreover, CyB induced zebrafish liver injury and increased hepatocyte apoptosis, which increased the protein expression levels of Bax, TLR4, NF-kB p65 and STAT3 in zebrafish. Finally, specific inhibition of TLR signaling pathway by TLR4 knock-down could significantly reduce the expression of inflammatory cytokines induced by CyB exposure. Taken together, these informations demonstrated that CyB could induce the hepatotoxicity and immunotoxicity in zebrafish embryos, and the expression levels of many genes involved in lipid metabolism and immune inflammation were obtained by RNA-Seq analysis. This study provides valuable information for future elucidating the aquatic toxicity of herbicide in aquatic ecosystems.

Pexidartinib hydrochloride exposure induces developmental toxicity and immunotoxicity in zebrafish embryos via activation of Wnt signaling.

Pexidartinib, a macrophage colony-stimulating factor receptor (CSF-1R) inhibitor, is indicated for the treatment of tendon sheath giant cell tumor (TGCT). However, few studies on the toxicity mechanisms of pexidartinib for embryonic development. In this study, the effects of pexidartinib on embryonic development and immunotoxicity in zebrafish were investigated. Zebrafish embryos at 6 h post fertilization (6 hpf) were exposed to 0, 0.5, 1.0, and 1.5 µM concentrations of pexidartinib, respectively. The results showed that different concentrations of pexidartinib induced the shorter body, decreased heart rate, reduced number of immune cells and increase of apoptotic cells. In addition, we also detected the expression of Wnt signaling pathway and inflammation-related genes, and found that these genes expression were significantly upregulated after pexidartinib treatment. To test the effects of embryonic development and immunotoxicity due to hyperactivation of Wnt signaling after pexidartinib treatment, we used IWR-1, Wnt inhibitor, for rescue. Results show that IWR-1 could not only rescue developmental defects and immune cell number, but also downregulate the high expression of Wnt signaling pathway and inflammation-related caused by pexidartinib. Collectively, our results suggest that pexidartinib induces the developmental toxicity and immunotoxicity in zebrafish embryos through hyperactivation of Wnt signaling, providing a certain reference for the new mechanisms of pexidartinib function.

Reactive oxygen signaling molecule inducible regulation of CRISPR-Cas9 gene editing.

We report development of a controllable gene editing tool that boronated gRNA, simply generated in situ, could regulate binding of gRNA molecules with either Cas9 endonuclease or target genes, thus serving as a modulator that can control CRISPR-Cas9 gene editing. Subsequent treatment with H2O2 facilitates the restoration of gene editing ability of the boronated gRNA to the level of using untreated gRNA. This is one of the few cases using small molecule to regulate CRISPR-Cas9 gene editing, which is a complement to the light approach, displaying great application potential. We develop a controllable gene editing tools based on the CRISPR-Cas9 gene editing system. This tool can be regulated by oxidative small molecule, i.e., H2O2. Compared with the light method, the application scope of our CRISPR-Cas9 systems have been widened with the small-molecule-triggered approaches, preventing the potential damage of cells or organism caused by UV light. In addition, the gain-of-function tools are expanding the gene code expansion for mechanistic studies of target enzymes since it provides a positive route to evaluate the activity of a given enzyme in dynamic and inversible regulation of targeting cellular processes.

Decoding selective auditory attention with EEG using a transformer model.

The human auditory system extracts valid information in noisy environments while ignoring other distractions, relying primarily on auditory attention. Studies have shown that the cerebral cortex responds differently to the sound source locations and that auditory attention is time-varying. In this work, we proposed a data-driven encoder-decoder architecture model for auditory attention detection (AAD), denoted as AAD-transformer. The model contains temporal self-attention and channel attention modules and could reconstruct the speech envelope by dynamically assigning weights according to the temporal self-attention and channel attention mechanisms of electroencephalogram (EEG). In addition, the model is conducted based on data-driven without additional preprocessing steps. The proposed model was validated using a binaural listening dataset, in which the speech stimulus was Mandarin, and compared with other models. The results showed that the decoding accuracy of the AAD-transformer in the 0.15-second decoding time window was 76.35%, which was much higher than the accuracy of the linear model using temporal response function in the 3-second decoding time window (increased by 16.27%). This work provides a novel auditory attention detection method, and the data-driven characteristic makes it convenient for neural-steered hearing devices, especially those who speak tonal languages.

An Automated Solid-Phase Extraction-UPLC-MS/MS Method for Simultaneous Determination of Sulfonamide Antimicrobials in Environmental Water.

The large-scale use of sulfonamide antimicrobials in human and veterinary medicine has seriously endangered the ecological environment and human health. The objective of this study was to develop and validate a simple and robust method for the simultaneous determination of seventeen sulfonamides in water using ultra-high performance liquid chromatography-tandem mass spectrometry coupled with fully automated solid-phase extraction. Seventeen isotope-labeled internal standards for sulfonamides were used to correct matrix effects. Several parameters affecting extraction efficiency were systematically optimized, and the enrichment factors were up to 982-1033 and only requiring about 60 min per six samples. Under the optimized conditions, this method manifested good linearity (0.05-100 µg/L), high sensitivity (detection limits: 0.01-0.05 ng/L), and satisfactory recoveries (79-118%) with acceptable relative standard deviations (0.3-14.5%, n = 5). The developed method can be successfully utilized for the determination of 17 sulfonamides in pure water, tap water, river water, and seawater. In total, six and seven sulfonamides were detected in river water and seawater, respectively, with a total concentration of 8.157-29.676 ng/L and 1.683-36.955 ng/L, respectively, and sulfamethoxazole was the predominant congener.

B16F10 Cell Membrane-Based Nanovesicles for Melanoma Therapy Are Superior to Hyaluronic Acid-Modified Nanocarriers.

Some cancer cell membrane (CCM)-derived nanovesicles show strong homing effects and are used for targeted cancer therapy. By co-constructing the B16F10 cell membrane with a PEGylated phospholipid membrane, a new nanocarrier with a composite nanocrown structure was developed, which can evade immune recognition and actively target homologous melanoma. The nanocrowns have an encapsulation efficiency of more than 90% for paclitaxel and showed no significant difference (p > 0.05) from the PEGylated phospholipid membrane vesicles. Compared with the hyaluronic acid-modified PEGylated phospholipid membrane vesicles, the biomimetic nanocrowns enhanced the escape of nanovesicles from reticuloendothelial cells in vitro and extended the circulation time in vivo; moreover, the nanocrowns showed superior melanoma-targeted drug delivery capability and improved anticancer effects of paclitaxel as demonstrated by the inhibition of B16F10 cell proliferation and induction of apoptosis by interfering with microtubule formation. In contrast, the modification of hyaluronic acid did not increase the targeting capacity or antitumor effects of the nanocrowns, confirming that the superior targeting capacity was mediated by the exposed homologous CCMs rather than by hyaluronic acid. Our results demonstrate the potential of using biomimetic nanocrowns for active melanoma-targeted therapy.

Identification of a new mutant allele of ZmYSL2 that regulates kernel development and nutritional quality in maize.

The discovery and characterization of the opaque endosperm gene provide ideas and resources for the production and application of maize. We found an o213 mutant whose phenotype was opaque and shrunken endosperm with semi-dwarf plant height. The protein, lipid, and starch contents in the o213 endosperm were significantly decreased, while the free amino acid content in the o213 endosperm significantly increased. The aspartic acid, asparagine, and lysine contents were raised in the o213 endosperm by 6.5-, 8.5-, and 1.7-fold, respectively. Genetic analysis showed that this o213 mutant is a recessive single-gene mutation. The position mapping indicated that o213 is located in a 468-kb region that contains 11 protein-encoding genes on the long arm of chromosome 5. The coding sequence analysis of candidate genes between the WT and o213 showed that ZmYSL2 had only a single-base substitution (A-G) in the fifth exon, which caused methionine substitution to valine. Sequence analysis and the allelic test showed that o213 is a new mutant allele of ZmYSL2. The qRT-PCR results indicated that o213 is highly expressed in the stalks and anthers. Subcellular localization studies showed that o213 is a membrane transporter. In the variation analysis of o213, the amplification of 65 inbred lines in GWAS showed that this 3-bp deletion of the first exon of o213 was found only in temperate inbred lines, implying that the gene was artificially affected in the selection process. Our results suggest that o213 is an important endosperm development gene and may serve as a genetic resource. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-022-01278-9.

Effects of supplementing with Humulus scandens on the growth performance and gut microbiota in piglets.

AIMS: This study was conducted to evaluate the effects of feeding Humulus scandens (Hu) on growth performance and gut microbiota in piglets. METHODS AND RESULTS: A total of 120 piglets were allocated to four dietary treatments (1) CON, basal diet; (2) T1, basal diet + 2.0% Hu; (3) T2, basal diet + 2.8% Hu and (4) T3, basal diet + 3.6% Hu. The results showed that dietary H. scandens supplementation increased the final body weight and average daily gain. Furthermore, H. scandens supplementation in T1 groups increased the content of total protein, globulin and IgG in serum and the apparent digestibility of crude protein. Gut microbiota analysis showed that H. scandens treatment in T1 groups increased the abundances of Lactobacillus, Ruminococcaceae, Enterococcus and Pseudomonas in cecum content. CONCLUSIONS: These findings suggested that dietary H. scandens supplementation improved the growth performance, immunological function and nutrient apparent digestibility as well as modulating the gut microbiota in piglets. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributed to developing new feed resources and might provide an alternative strategy for growth promotion in piglets.

Development and evaluation of the Cantonese matrix sentence test.

OBJECTIVE: To develop the Cantonese matrix (YUEmatrix) test according to the international standard procedure and examine possible different outcomes in another tonal language. DESIGN: A 50-word Cantonese base-matrix was established. Word-specific speech recognition functions, speech recognition thresholds (SRT), and slopes were obtained. The speech material was homogenised in intelligibility by applying level corrections up to ± 3 dB. Subsequently, the YUEmatrix test was evaluated in five aspects: training effect, test-list equivalence, test-retest reliability, establishment of reference data for normal-hearing Cantonese-speakers, and comparison with the Cantonese-Hearing-In-Noise-Test. STUDY SAMPLE: Overall, 64 normal-hearing native Cantonese-speaking listeners. RESULTS: SRT measurements with adaptive procedures resulted in a reference SRT of -9.7 ± 0.7 dB SNR for open-set and -11.1 ± 1.2 dB SNR for the closed-set response format. Fixed SNR measurements suggested a test-specific speech intelligibility function slope of 15.5 ± 0.7%/dB. Seventeen 10-sentences base test lists were confirmed to be equivalent with respect to speech intelligibility. Training effect was not observed after two measurements of 20-sentences lists. CONCLUSIONS: The YUEmatrix yields comparable results to matrix tests in other languages including Mandarin. Level adjustments to homogenise sentences appear to be less effective for tonal languages than for most other languages developed so far.

Removal of Copper(II) from Aqueous Environment Using Silk Sericin-Derived Carbon.

Sericin is a by-product of the silk industry. Its recycling contributes to environmental protection and the sustainable development of the cocoon silk industry. In this paper, on the basis of realizing sericin enrichment in solution, the Cu(II) adsorption capacities of sericin-derived carbon (SC), prepared at different pyrolysis temperatures, were studied. SC was characterized using scanning electron microscopy (SEM) and the zeta potential. The effects of the initial concentration of Cu(II), pH, adsorption temperature, and contact time on the adsorption process were evaluated, followed by an investigation of the mechanism of Cu(II) adsorption by SC. The results showed that SC has a porous structure that provides sites for Cu(II) adsorption. The maximum adsorption capacity of Cu(II) onto SC1050, 17.97 mg/g, was obtained at an adsorption temperature of 35 °C and a pH of 5.5. In addition, the pseudo-second-order kinetic model and Langmuir isotherm model correctly described the adsorption process of Cu(II) onto SC1050. Therefore, SC can act as a potential adsorbent for removing Cu(II) from water. This study helps promote the effective use of cocoon silk resources.

Silk Sericin Enrichment through Electrodeposition and Carbonous Materials for the Removal of Methylene Blue from Aqueous Solution.

The recycling and reuse of biomass waste for the preparation of carbon-based adsorbents is a sustainable development strategy that has a positive environmental impact. It is well known that a large amount of silk sericin (SS) is dissolved in the wastewater from the silk industry. Utilizing the SS instead of discharging it into the environment without further treatment would reduce environmental and ecological problems. However, effective enrichment of the SS from the aqueous solution is a challenge. Here, with the help of carboxymethyl chitosan (CMCS), which can form a gel structure under low voltage, an SS/CMCS hydrogel with SS as the major component was prepared via electrodeposition at a 3 V direct-current (DC) voltage for five minutes. Following a carbonization process, an SS-based adsorbent with good performance for the removal of methylene blue (MB) from an aqueous solution was prepared. Our results reveal that the SS/CMCS hydrogel maintains a porous architecture before and after carbonization. Such structure provides abundant adsorption sites facilitating the adsorption of MB molecules, with a maximum adsorptive capacity of 231.79 mg/g. In addition, it suggests that the adsorption is an exothermic process, has a good fit with the Langmuir model, and follows the intra-particle diffusion model. The presented work provides an economical and feasible path for the treatment of wastewater from dyeing and printing.

Incidence and clinical features of acute multiple small cerebellar infarction.

OBJECTIVE: To investigate the clinical and imaging features and to identify possible etiology of acute multiple small cerebellar infarction (MSCI). METHODS: We retrospectively enrolled 220 patients with acute cerebellar infarction, divided them into MSCI and large cerebellar infarction (LCI) groups, according to the quantity and size of lesions confirmed by MRI analysis. Clinical and imaging features were compared between the two groups to explore the possible etiology and pathogenesis. RESULTS: Among 220 patients, 90 patients presented MSCI symptoms. The proportions of extracerebellar lesions (P = 0.001) and bilateral infarction (P = 0.001) in the MSCI group were higher than those in the LCI group. No significant differences were found in terms of age, gender, and common vascular risk factors between the two groups. The proportions of vertigo and headache in the MSCI group were significantly lower than those in the LCI group (P < 0.000 and 0.034, respectively), and limb weakness was significantly higher (P = 0.039) in the MSCI patients. Moreover, the proportions of nystagmus and ataxia in the MSCI group were significantly lower than those in the LCI group (P < 0.043 and 0.003, respectively). The MSCI group had higher proportions of ACA and MCA stenosis, while the proportion of posterior circulation stenosis was similar between the two groups. Infarctions involving the posterior inferior cerebellar (PICA) region and mixed territories were far more frequent than those involving the anterior inferior cerebellar artery (AICA) region and superior cerebellar artery (SCA) territory (P < 0.05). Large-artery atherosclerosis and multiple plus undetermined etiology were the main etiological factors of MSCI. CONCLUSION: In patients with acute cerebellar infarction, 30% of patients presented with MSCI. MSCI and LCI showed similar vascular risk factors and vascular stenosis in the posterior circulation system. Patients with MSCI should pay more attention to evaluating anterior circulation vessels' anatomy. Large-artery atherosclerosis was the main pathogenesis of acute MSCI. Assessment of cerebral vessels might be critically required in patients with MSCI complicated atrial fibrillation.

Occurrence of Free-Form and Conjugated Bisphenol Analogues in Marine Organisms.

Upon exposure, most bisphenol analogues (BPs) are rapidly metabolized to BP conjugates in organisms. Monitoring studies on BPs in aquatic organisms have been mainly focused on free-form BPs. However, the relative amount of conjugated BPs in organisms is still not well known, especially in marine organisms. In this study, we collected marine organisms (13 species; n = 74), as well as seawater (n = 15), from East China Sea, and analyzed them for bisphenol A (BPA) and its eight analogues. In seawater, BPA was the predominant BP (mean ± standard deviation (SD), 18 ± 9.7 ng/L), followed by bisphenol S (BPS; 3.7 ± 2.8 ng/L), bisphenol F (BPF; 0.31 ± 0.17 ng/L), and bisphenol AF (BPAF; 0.24 ± 0.15 ng/L). The whole body of each marine organism was used for BP analysis in this study. BPA (mean 3.8 ng/g, range 1.2-7.7 ng/g) and BPS (1.5 ng/g, 0.19-6.1 ng/g) were still the predominant BPs in marine organisms without hydrolysis. After enzymatic hydrolysis treatment, mean concentrations of BPs increased 1.8 (BPS)-3.7 (BPA) times in marine organisms. Correspondingly, on average, 74, 52, 49, 48, and 45% of BPA, bisphenol Z, BPF, BPS, and BPAF were present in conjugated forms, respectively, in marine organisms. Moreover, bioaccumulation and biomagnification of BPs were evaluated using pooled marine organism samples. We first found that the mean log BAF values of BPs increased 0.091 (BPAF) to 0.31 (BPA) times in marine organisms after hydrolysis. Overall, this study first determines the amount of BPs present in conjugated forms in marine organisms, which are helpful for better understanding the occurrence of BPs in organisms.

Determination of polyhalogenated carbazoles in waters at low nanogram-per-liter concentrations with solid-phase disk extraction.

Polyhalogenated carbazoles, a class of emerging contaminants with persistence and dioxin-like toxicity, have received increasing attention in recent years. In this study, a simple, rapid, sensitive, and high throughput method based on solid-phase disk extraction and gas chromatography-mass spectrometry was described for the determination of polyhalogenated carbazoles in low nanogram-per-liter range in water samples. The proposed solid-phase disk extraction method was initially optimized, and the optimum experimental conditions found were 1 L water sample (pH 6-9) extracted and enriched by Empore 3-stn octadecyl disk at flow rate of 5 to 50 mL/min and eluted by 5 mL of acetone and 3 × 10 mL methylene dichloride. The linearity of the method ranged from 0.2 to 50 ng/L for carbazole and 11 polyhalogenated carbazoles, with correlation coefficients ranging from 0.9951 to 0.9996. The limits of detection were in the low nanogram per liter level, ranging from 0.018 to 0.12 ng/L. Finally, the optimized method was applied for determining trace levels of carbazole and 11 polyhalogenated carbazoles in tap water and seawater samples with good recovery of 86.6-112.8%. Carbazole and 3-7 polyhalogenated carbazoles were detected, and 3,6-dichlorocarbazole was the predominant congener both in tap water and seawater.

Synthesis and evaluation of antimicrobial and anticancer activities of 3-phenyl-1-phenylsulfonyl pyrazoles containing an aminoguanidine moiety.

A series of 3-phenyl-1-phenylsulfonyl pyrazoles containing an aminoguanidine moiety was designed, synthesized, and evaluated for their antimicrobial and anticancer activities. The majority of the target compounds showed broad-spectrum antimicrobial activity against the tested strains, with minimum inhibitory concentration (MIC) values ranging from 2 to 64 µg/ml. Compound 5k, showing the most potent antimicrobial activity against Bacillus subtilis CMCC 63501 and multidrug-resistant Staphylococcus aureus ATCC 43300 with an MIC value of 2 µg/ml, was the most promising one in this series. It was also effective for S. aureus ATCC 33591 and multidrug-resistant Escherichia coli ATCC BAA-196 at higher concentrations. The bactericidal time-kill kinetics test illustrated that compound 5k had rapid bactericidal potential. Docking results exhibited that compound 5k showed various kinds of binding to the FabH receptor, reflecting that 5k could bind with the active site well. All compounds showed excellent activity against the investigated cancer cells, with IC50 values ranging from 1.90 to 54.53 µM. Among them, compound 5f showed prominent cytotoxicity with IC50 = 1.90 µM against A549 cells, while exhibiting lower inhibitory activity against 293T cells (IC50 = 41.72 µM), indicating that it has the potential for a good therapeutic index as an anticancer drug.

DNA methylation and histone acetylation are involved in Wnt10b expression during the secondary hair follicle cycle in Angora rabbits.

Secondary hair follicles (SHFs) in the Angora rabbit exhibit classic cyclic hair development, but the multiple molecular signals involved in hair cycling are yet to be explored in detail. In the present study, we investigated the expression pattern, methylation and histone H3 acetylation status of Wnt10b, as a molecular signal participating in hair cycling, during the SHF cycle in the Angora rabbit. Expression of Wnt10b at the anagen phase was significantly higher than that at both the telogen and catagen phases, suggesting that Wnt10b might serve as a critical activator during cyclic transition of SHFs. Methylation frequency of the fifth CpG site (CpG5-175 bp) in CpG islands at the anagen phase was lower than that at both the catagen and telogen phases. The methylation status of the CpG5 site was negatively correlated with Wnt10b expression. This indicated that the methylation of CpG5 might participate in Wnt10b transcriptional suppression in SHFs. Furthermore, histone H3 acetylation status in the regions-256~-11 bp and 98 ~ 361 bp were significantly lower at both the catagen and telogen phases than at the anagen phase. The histone H3 acetylation level was significantly positively correlated with Wnt10b expression. This confirmed that histone acetylation was likely involved in upregulating Wnt10b transcription in SHFs. Additionally, potential binding to the transcription factors ZF57 and HDBP was predicted within the CpG5 site. In conclusion, our findings reveal the epigenetic mechanism of Wnt10b transcription and provide a new insight into epigenetic regulation during the SHF cycle in the Angora rabbit.

A comparison of methods for effective differentiation of the frozen-thawed 3T3-L1 cells.

The differentiation efficiency of 3T3-L1 preadipocytes is an essential factor affecting studies on cellular mechanisms associated with obesity, diabetes, and related disorders. Differentiation of 3T3-L1 cells is commonly induced by an adipogenic cocktail containing insulin, dexamethasone (DEX), and 3-isobutyl-1-methylxanthine (IBMX). However, 3T3-L1 cells after freezing and thawing for many times always have a low differentiation efficiency. To solve this problem, we compared the differentiation efficiency of six commonly used adipogenic cocktails and protocols published in 2017. On this basis, we further compared 18 adipogenic cocktails with 2 µM rosiglitazone added and/or with a prolonged treatment with IBMX. The results revealed that the adipogenic cocktail containing 0.5 mM IBMX, 1 µM DEX, and 10 µg/mL insulin was the most effective for the frozen-thawed 3T3-L1 cells differentiation. Rosiglitazone, and IBMX under a prolonged treatment, could improve the differentiation efficiency of the frozen-thawed 3T3-L1 cells. However, the effect was closely related to concentrations of agents in the adipogenic cocktails.