Comparison of capture-based mtDNA sequencing performance between MGI and illumina sequencing platforms in various sample types.

BACKGROUND: Mitochondrial genome abnormalities can lead to mitochondrial dysfunction, which in turn affects cellular biology and is closely associated with the development of various diseases. The demand for mitochondrial DNA (mtDNA) sequencing has been increasing, and Illumina and MGI are two commonly used sequencing platforms for capture-based mtDNA sequencing. However, there is currently no systematic comparison of mtDNA sequencing performance between these two platforms. To address this gap, we compared the performance of capture-based mtDNA sequencing between Illumina's NovaSeq 6000 and MGI's DNBSEQ-T7 using tissue, peripheral blood mononuclear cell (PBMC), formalin-fixed paraffin-embedded (FFPE) tissue, plasma, and urine samples. RESULTS: Our analysis indicated a high degree of consistency between the two platforms in terms of sequencing quality, GC content, and coverage. In terms of data output, DNBSEQ-T7 showed higher rates of clean data and duplication compared to NovaSeq 6000. Conversely, the amount of mtDNA data obtained by per gigabyte sequencing data was significantly lower in DNBSEQ-T7 compared to NovaSeq 6000. In terms of detection mtDNA copy number, both platforms exhibited good consistency in all sample types. When it comes to detection of mtDNA mutations in tissue, FFPE, and PBMC samples, the two platforms also showed good consistency. However, when detecting mtDNA mutations in plasma and urine samples, significant differenceof themutation number detected was observed between the two platforms. For mtDNA sequencing of plasma and urine samples, a wider range of DNA fragment size distribution was found in NovaSeq 6000 when compared to DNBSEQ-T7. Additionally, two platforms exhibited different characteristics of mtDNA fragment end preference. CONCLUSIONS: In summary, the two platforms generally showed good consistency in capture-based mtDNA sequencing. However, it is necessary to consider the data preferences generated by two sequencing platforms when plasma and urine samples were analyzed.

Cyclophosphamide activates ferroptosis-induced dysfunction of Leydig cells via SMAD2 pathway†.

Cyclophosphamide (CP) is a widely used chemotherapeutic drug and immunosuppressant in the clinic, and the hypoandrogenism caused by CP is receiving more attention. Some studies found that ferroptosis is a new mechanism of cell death closely related to chemotherapeutic drugs and plays a key role in regulating reproductive injuries. The purpose of this study is to explore ferroptosis' role in testicular Leydig cell dysfunction and molecular mechanisms relating to it. In this study, the level of ferroptosis in the mouse model of testicular Leydig cell dysfunction induced by CP was significantly increased and further affected testosterone synthesis. The ferroptosis inhibitors ferrostatin-1 (Fer-1) and iron chelator deferoxamine (DFO) can improve injury induced by CP. The results of immunohistochemistry showed that Fer-1 and DFO could improve the structural disorder of seminiferous tubules and the decrease of the number of Leydig cells in testicular tissue induced by CP. Immunofluorescence and western blot confirmed that Fer-1 and DFO could improve the expression of key enzymes in testosterone synthesis. The activation of SMAD family member 2 (Smad2)/cyclin-dependent kinase inhibitor 1A (Cdkn1a) pathway can improve the ferroptosis of Leydig cells induced by CP and protect the function of Leydig cells. By inhibiting the Smad2/Cdkn1a signal pathway, CP can regulate ferroptosis, resulting in testicular Leydig cell dysfunction. In this study, CP-induced hypoandrogenism is explained theoretically and a potential therapeutic strategy is provided.

High-intensity interval training ameliorates postnatal immune activation-induced mood disorders through KDM6B-regulated glial activation.

Postnatal immune activation (PIA) induces persistent glial activation in the brain and causes various neuropathologies in adults. Exercise training improves stress-related mood disorders; however, the role of exercise in psychiatric disorders induced by early-life immune activation and the association between exercise training and glial activation remain unclear. We compared the effects of different exercise intensities on the PIA model, including high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT). Both HIIT and MICT in adolescent mice inhibited neuroinflammation, remodeled synaptic plasticity, and improved PIA-induced mood disorders in adulthood. Importantly, HIIT was superior to MICT in terms of reducing inflammation and increasing body weight. RNA-seq of prefrontal cortex (PFC) tissues revealed a gene expression pattern, confirming that HIIT was more effective than MICT in improving brain glial cell activation through epigenetic modifications of KDM6B. We investigated the role of KDM6B, a specific histone lysine demethylation enzyme - histone 3 lysine 27 demethylase, in inhibiting glial activation against PIA-induced depression and anxiety by regulating the expression of IL-4 and brain-derived neurotrophic factor (BDNF). Overall, our data support the idea that HIIT improves PIA-induced mood disorders by regulating KDM6B-mediated epigenetic mechanisms and indicate that HIIT might be superior to MICT in improving mood disorders with PIA in mice. Our findings provide new insights into the treatment of anxiety and depression disorders.

Association Between Peritoneal Glucose Absorption, Lipid Metabolism, and Cardiovascular Disease Risk in Nondiabetic Patients on Peritoneal Dialysis.

BACKGROUND: Excessive sugar intake increases the energy metabolic burden and the risk of cardiovascular disease (CVD). Patients on peritoneal dialysis absorb much more glucose than the World Health Organization recommends, but the link to CVD is unclear. OBJECTIVE: To identify the association between peritoneal glucose absorption, lipid metabolism, and CVD. METHODS: We applied generalized additive mixed effects and mixed effects Cox proportional hazard models to evaluate the impact of peritoneal glucose absorption on lipid profiles and CVD risk. We performed subgroup analyses by using protein intake (normalized protein nitrogen appearance [nPNA] and normalized protein catabolic rate [nPCR] were used to assess protein intake) and high-sensitivity C-reactive protein (hs-CRP). RESULTS: After multivariable adjustment, peritoneal glucose absorption per 10 g/d increase was associated with an increase in cholesterol of 0.145 (95% confidence interval [CI]: 0.086-0.204) mmol/L. No link with the total risk of CVD was observed; however, protein intake and hs-CRP levels affected the relationship between glucose absorption and CVD risk. Patients with values for nPNA and nPCR <1.0 g/(kg·d) were associated with a lower risk of CVD (hazard ratio [HR] 95% CI: 0.68 (0.46-0.98)) with glucose absorption per 10 g/d increase. While patients with hs-CRP levels ≥3 mg/d or values for nPNA or nPCR ≥1.0 g/(kg·d) were associated with a higher risk of CVD (HR 95% CI: 1.32 (1.07-1.63); 1.31 (1.02-1.68)) for glucose absorption per 10 g/d increase. CONCLUSIONS: Our study found a positive correlation between peritoneal glucose absorption and lipid profiles. Increased glucose absorption was associated with a lower risk of CVD in lower protein intake patients and a higher risk of CVD in higher hs-CRP or protein intake levels in patients on peritoneal dialysis.

Melatonin improves mouse oocyte quality from 2-ethylhexyl diphenyl phosphate-induced toxicity by enhancing mitochondrial function.

2-Ethylhexyl diphenyl phosphate (EHDPP) is a representative organophosphorus flame retardant (OPFR) that has garnered attention due to its widespread use and potential adverse effects. EHDPP exhibits cytotoxicity, genotoxicity, developmental toxicity, and endocrine disruption. However, the toxicity of EHDPP in mammalian oocytes and the underlying mechanisms remain poorly understood. Melatonin is a natural free radical scavenger that has demonstrated cytoprotective properties. In this study, we investigated the effect of EHDPP on mouse oocytes in vitro culture system and evaluated the rescue effect of melatonin on oocytes exposed to EHDPP. Our results indicated that EHDPP disrupted oocyte maturation, resulting in the majority of oocytes arrested at the metaphase I (MI) stage, accompanied by cytoskeletal damage and elevated levels of reactive oxygen species (ROS). Nevertheless, melatonin supplementation partially rescued EHDPP-induced mouse oocyte maturation impairment. Results of single-cell RNA sequencing (scRNA-seq) analysis elucidated potential mechanisms underlying these protective effects. According to the results of scRNA-seq, we conducted further tests and found that EHDPP primarily disrupts mitochondrial distribution and function, kinetochore-microtubule (K-MT) attachment, DNA damage, apoptosis, and histone modification, which were rescued upon the supplementation of melatonin. This study reveals the mechanisms of EHDPP on female reproduction and indicates the efficacy of melatonin as a therapeutic intervention for EHDPP-induced defects in mouse oocytes.

Nicotinamide mononucleotide maintains cytoskeletal stability and fortifies mitochondrial function to mitigate oocyte damage induced by Triocresyl phosphate.

Triocresyl phosphate (TOCP) was commonly used as flame retardant, plasticizer, lubricant, and jet fuel additive. Studies have shown adverse effects of TOCP on the reproductive system. However, the potential harm brought by TOCP, especially to mammalian female reproductive cells, remains a mystery. In this study, we employed an in vitro model for the first time to investigate the effects of TOCP on the maturation process of mouse oocytes. TOCP exposure hampered the meiotic division process, as evidenced by a reduction in the extrusion of the first polar body from oocytes. Subsequent research revealed the disruption of the oocyte cell cytoskeleton induced by TOCP, resulting in abnormalities in spindle organization, chromosome alignment, and actin filament distribution. This disturbance further extended to the rearrangement of organelles within oocytes, particularly affecting the mitochondria. Importantly, after TOCP treatment, mitochondrial function in oocytes was impaired, leading to oxidative stress, DNA damage, cell apoptosis, and subsequent changes of epigenetic modifications. Supplementation with nicotinamide mononucleotide (NMN) alleviated the harmful effects of TOCP. NMN exerted its mitigating effects through two fundamental mechanisms. On one hand, NMN conferred stability to the cell cytoskeleton, thereby supporting nuclear maturation. On the other hand, NMN enhanced mitochondrial function within oocytes, reducing the excess reactive oxygen species (ROS), restoring meiotic division abnormalities caused by TOCP, preventing oocyte DNA damage, and suppressing epigenetic changes. These findings not only enhance our understanding of the molecular basis of TOCP induced oocyte damage but also offer a promising avenue for the potential application of NMN in optimizing reproductive treatment strategies.

Generation of a Hydrophobic Protrusion on Nanoparticles to Improve the Membrane-Anchoring Ability and Cellular Internalization.

Controlling the nanoparticle-cell membrane interaction to achieve easy and fast membrane anchoring and cellular internalization is of great importance in a variety of biomedical applications. Here we report a simple and versatile strategy to maneuver the nanoparticle-cell membrane interaction by creating a tunable hydrophobic protrusion on Janus particles through swelling-induced symmetry breaking. When the Janus particle contacts cell membrane, the protrusion will induce membrane wrapping, leading the particles to docking to the membrane, followed by drawing the whole particles into the cell. The efficiencies of both membrane anchoring and cellular internalization can be promoted by optimizing the size of the protrusion. In vitro, the Janus particles can quickly anchor to the cell membrane in 1 h and be internalized within 24 h, regardless of the types of cells involved. In vivo, the Janus particles can effectively anchor to the brain and skin tissues to provide a high retention in these tissues after intracerebroventricular, intrahippocampal, or subcutaneous injection. This strategy involving the creation of a hydrophobic protrusion on Janus particles to tune the cell-membrane interaction holds great potential in nanoparticle-based biomedical applications.

Mitochondrial DNA haplogroup M7: A predictor of poor prognosis for colorectal cancer patients in Chinese population.

Haplogroups and single-nucleotide polymorphisms (SNP) of mitochondrial DNA (mtDNA) were associated with the prognosis of many types of cancer patients. However, whether mtDNA haplogroups contribute to clinical outcomes of colorectal cancer (CRC) in Chinese population remains to be determined. In this study, mtDNA of tissue samples from 445 CRC patients from Northwestern China was sequenced to evaluate the association between haplogroup and prognosis. The mtDNA sequencing data of 1015 CRC patients from Southern China were collected for validation. We found patients with mtDNA haplogroup M7 had a significantly higher death risk when compared with patients with other haplogroups in both Northwestern (Hazard ratio [HR] = 3.093, 95% CI = 1.768-5.411, p < 0.001) and Southern (HR = 1.607, 95% CI = 1.050-2.459, p = 0.029) China. Then, a haplogroup M7-based mtSNP classifier was selected by using LASSO Cox regression analysis. A nomogram comprising the mtSNP classifier and clinicopathological variables was developed to predict the prognosis of CRC patients (area under the curve [AUC] 0.735, 95% CI = 0.679-0.791). Furthermore, patients with high- and low-risk scores calculated by the haplogroup M7-based mtSNP classifier exhibited significantly different overall survival (OS) and recurrence-free survival (RFS) (all p < 0.001). Finally, RNA-seq and immunohistochemical analyses indicated the poor prognosis of patients with haplogroup M7 may be related to mitochondrial dysfunction and immune abnormalities in CRC tissues. In conclusion, the haplogroup M7 and haplogroup M7-based mtSNP classifier seems to be a practical and reliable prognostic predictor for CRC patients, which provides a potential tool of clinical decision-making for patients with haplogroup M7 in Chinese population.

A new method for calculating interplanar spacing to distinguish between similar phases in EBSD.

Commercial electron backscatter diffraction (EBSD) systems generally use interplanar angle matching for pattern indexing, and thus, they are unable to distinguish between some similar phases with close interplanar angles, such as Al and Si. The interplanar spacing is more diagnostic but generally difficult to apply in pattern indexing because it lacks precision. In this study, we proposed an efficient approach for accurately measuring interplanar spacing by correcting the reciprocal-lattice vector (RLV). The phase discrimination of Al and Si was performed by interplanar spacing matching. The Kikuchi bands were identified automatically by the self-developed method using pattern rotation combined with grey gradient recognition without the help of human eyes. The reliable RLV relationship was extracted by accurately drawing reciprocal-lattice vectors. The lengths of RLVs were corrected, and then the RLVs were used for evaluating lattice spacing. The results of five Kikuchi patterns with different clarity showed that this new method reduced the average error of interplanar spacings by 50.611% and achieved an average accuracy of 1.644% for lattice spacing calculation. The method could distinguish structures with a difference in lattice spacing of at least 3.3%. This method was also effective for fuzzy patterns and partially missing Kikuchi bands and might be used as a new strategy for improving the calculation accuracy of lattice spacing for fuzzy patterns. The method did not have additional requirements concerning the number of detected Kikuchi bands and poles. The accuracy of lattice spacing could be effectively improved by correcting the RLVs based on routine pattern recognition. This method might be used as an auxiliary approach to differentiate between similar phases and is well-adapted to the existing commercial EBSD system.

Cyclic Peptides from the Opportunistic Pathogen Basidiobolus meristosporus.

Three new cyclic peptides, meristosporins A, B, and C (1-3), one of which features an unusual amino acid, were isolated from the opportunistic pathogen Basidiobolus meristosporus and identified by 1D, 2D NMR, MS/MS, and Marfey's analysis. The biosynthetic pathway of the hexapeptide meristosporin A (1) was deduced based on nonribosomal peptide synthetase gene clusters analysis. Compounds 1 and 2 showed cytotoxicity to RAW264.7 and 293T cells, respectively. These compounds may be involved in the fungal injury caused to human cells.

Shear banding in monodisperse polymer melt.

We performed a series of molecular dynamics simulations on monodisperse polymer melts to investigate the formation of shear banding. Under high shear rates, shear banding occurs, which is intimately accompanied by the entanglement heterogeneity. Interestingly, the same linear relationship between the end-to-end distance Ree and entanglement density Z is observed at homogeneous flow before the onset of shear banding and at the shear banding state, where Ree ∼ ln(Wi0.87)-ξ0Z is proposed as the criterion to describe the dynamic force balance of the molecular chain in flow with a high rate. Deviating from this relation leads to a force imbalance and results in the emergence of shear banding. We establish a scaling relation between the disentanglement rate Vd and the Weissenberg number Wi as Vd∼Wi0.87 for stable flow in homogeneous shear and shear banding states. The formation of shear banding prevents chains from further stretching and disentanglement. The transition from homogeneous shear to shear banding partially dissipates the increased free energy from shear and reduces the free energy of the system.

Dynamic Metal Nanoclusters: A Review on Accurate Crystal Structures.

Dynamic metal nanoclusters have garnered widespread attention due to their unique properties and potential applications in various fields. Researchers have been dedicated to developing new synthesis methods and strategies to control the morphologies, compositions, and structures of metal nanoclusters. Through optimized synthesis methods, it is possible to prepare clusters with precise sizes and shapes, providing a solid foundation for subsequent research. Accurate determination of their crystal structures is crucial for understanding their behavior and designing custom functional materials. Dynamic metal nanoclusters also demonstrate potential applications in catalysis and optoelectronics. By manipulating the sizes, compositions, and surface structures of the clusters, efficient catalysts and optoelectronic materials can be designed and synthesized for various chemical reactions and energy conversion processes. This review summarizes the research progress in the synthesis methods, crystal structure characterization, and potential applications of dynamic metal nanoclusters. Various nanoclusters composed of different metal elements are introduced, and their potential applications in catalysis, optics, electronics, and energy storage are discussed. Additionally, the important role of dynamic metal nanoclusters in materials science and nanotechnology is explored, along with an overview of the future directions and challenges in this field.

Apparent mineralocorticoid excess: comprehensive overview of molecular genetics.

Apparent mineralocorticoid excess is an autosomal recessive form of monogenic disease characterized by juvenile resistant low-renin hypertension, marked hypokalemic alkalosis, low aldosterone levels, and high ratios of cortisol to cortisone metabolites. It is caused by defects in the HSD11B2 gene, encoding the enzyme 11ß-hydroxysteroid dehydrogenase type 2 (11ß-HSD2), which is primarily involved in the peripheral conversion of cortisol to cortisone. To date, over 50 deleterious HSD11B2 mutations have been identified worldwide. Multiple molecular mechanisms function in the lowering of 11ß-HSD2 activity, including damaging protein stability, lowered affinity for the substrate and cofactor, and disrupting the dimer interface. Genetic polymorphism, environmental factors as well as epigenetic modifications may also offer an implicit explanation for the molecular pathogenesis of AME. A precise diagnosis depends on genetic testing, which allows for early and specific management to avoid the morbidity and mortality from target organ damage. In this review, we provide insights into the molecular genetics of classic and non-classic apparent mineralocorticoid excess and aim to offer a comprehensive overview of this monogenic disease.

Complete genome sequence of a novel victorivirus infecting cicada flower (Cordyceps chanhua).

Cicada flower, Cordyceps chanhua, is a precious edible and medicinal mushroom with uses in both medicine and food in China. In this study, Cordyceps chanhua strain RCEF5995 was found to be coinfected by a previously characterized alternavirus, Cordyceps chanhua alternavirus 1 (CcAV1), and a novel victorivirus, tentatively named "Cordyceps chanhua victorivirus 1" (CcV1). Molecular characterization of CcV1 showed that its complete genome is 5,232 nucleotides long with a GC content of 57.5%. Sequence analysis indicated that CcV1 contains two overlapping open reading frames (ORFs), ORF1 and ORF2, encoding a putative coat protein (CP) of 742 amino acids (aa) and a putative RNA-dependent RNA polymerase (RdRp) of 836 aa, respectively. The termination codon of the CP ORF overlaps with the initiation codon of the RdRp ORF at the tetranucleotide sequence AUGA. Homolog searches, sequence comparisons, and phylogenetic analysis based on deduced amino acid sequences of RdRp indicated that CcV1 is a new member of the genus Victorivirus, family Totiviridae.

Mechanobiological responses of astrocytes in optic nerve head due to biaxial stretch.

BACKGROUND: Elevated intraocular pressure (IOP) is the main risk factor for glaucoma, which might cause the activation of astrocytes in optic nerve head. To determine the effect of mechanical stretch on the astrocytes, we investigated the changes in cell phenotype, proteins of interest and signaling pathways under biaxial stretch. METHOD: The cultured astrocytes in rat optic nerve head were stretched biaxially by 10 and 17% for 24 h, respectively. Then, we detected the morphology, proliferation and apoptosis of the stretched cells, and performed proteomics analysis. Protein expression was analyzed by Isobaric tags for relative and absolute quantification (iTRAQ) mass spectrometry. Proteins of interest and signaling pathways were screened using Gene Ontology enrichment analysis and pathway enrichment analysis, and the results were verified by western blot and the gene-chip data from Gene Expression Omnibus (GEO) database. RESULT: The results showed that rearrangement of the actin cytoskeleton in response to stimulation by mechanical stress and proliferation rate of astrocytes decreased under 10 and 17% stretch condition, while there was no significant difference on the apoptosis rate of astrocytes in both groups. In the iTRAQ quantitative experiment, there were 141 differential proteins in the 10% stretch group and 140 differential proteins in the 17% stretch group. These proteins include low-density lipoprotein receptor-related protein (LRP6), caspase recruitment domain family, member 10 (CARD10), thrombospondin 1 (THBS1) and tetraspanin (CD81). The western blot results of LRP6, THBS1 and CD81 were consistent with that of iTRAQ experiment. ANTXR2 and CARD10 were both differentially expressed in the mass spectrometry results and GEO database. We also screened out the signaling pathways associated with astrocyte activation, including Wnt/ß-catenin pathway, NF-κB signaling pathway, PI3K-Akt signaling pathway, MAPK signaling pathway, Jak-STAT signaling pathway, ECM-receptor interaction, and transforming growth factor-ß (TGF-ß) signaling pathway. CONCLUSION: Mechanical stimulation can induce changes in cell phenotype, some proteins and signaling pathways, which might be associated with astrocyte activation. These proteins and signaling pathways may help us have a better understanding on the activation of astrocytes and the role astrocyte activation played in glaucomatous optic neuropathy.

[Bone marrow mesenchymal stem cell-derived exosomes protect TM3 Leydig cells from cyclophosphamide-induced injury].

OBJECTIVE: To investigate the effect of exosomes derived from mouse bone marrow mesenchymal stem cells (BMSC) on the injury of TM3 Leydig cells induced by cyclophosphamide (CP). METHODS: The exosomes from BMSCs were extracted by ultrahigh speed centrifugation, and their particle size and morphology observed under the electron microscope, and their typical marker proteins examined by Western blot. The uptake of exosomes by TM3 Leydig cells was observed by co-culturing the exosomes with the TM3 cells. The viability and apoptosis rate of the TM3 cells in the normal control, CP-induction and CP+exosomes groups were detected using the CCK-8 method and flow cytometry respectively. ELISA was used to measure the testosterone (T) level in the cell supernatant, and Western blot adopted to determine the expression level of the steroidogenic acute regulatory (StAR) protein, a key enzyme related to T synthesis. RESULTS: The viability of the TM3 Leydig cells was markedly decreased and the apoptosis rate of the cells remarkably increased in the CP-induction group compared with that in the normal control, but both significantly restored after co-culture with exosomes (P < 0.01 and P < 0.05). The T level in the supernatant and the expression of the StAR protein in the cells were lower in the CP-induction than in the normal control group, but both dramatically increased in the CP+exosomes group (P < 0.01). CONCLUSION: Exosomes from BMSCs and protect TM3 Leydig cells from cyclophosphamide-induced injury and restore the level of testosterone secreted by the TM3 cells to a certain extent.

Polysaccharides extracted from balanophora polyandra Griff (BPP) ameliorate renal Fibrosis and EMT via inhibiting the Hedgehog pathway.

Renal interstitial fibrosis (RIF) is a crucial pathological change leading to chronic kidney disease (CKD). Currently, no effective medicines have been available for treating it. In our research, we examined the effects of polysaccharides extracted from Balanophora polyandra Griff (BPPs) on kidney fibrosis and epithelial to mesenchymal transition (EMT) in vivo and in vitro, aiming to explore the underlying mechanisms. By using the mice with unilateral urethral obstruction (UUO) as experimental subjects, we examined the medicinal values of BPPs on alleviating RIF. The effects of BPPs were evaluated by examining the histological staining and relative mRNA and protein expression levels of the related genes. The possible underlying mechanisms were further explored with human normal renal proximal tubular epithelia (HK-2 cells) as in vitro model. In UUO mice, BPP treatment could significantly alleviate interstitial fibrosis through reducing the components (Collagens I, III and IV) of extracellular matrix (ECM), and reducing the activation of fibroblasts producing these components, as revealed by inhibiting the hallmarks (fibronectin and α-SMA) of fibroblast activation. Furthermore, BPP administration increased the expression levels of matrix metalloproteinases (MMPs) and declined those of tissue inhibitors of metalloproteinases (TIMPs). BPPs markedly ameliorated EMT in both the kidneys of UUO mice and TGF-ß1 treated HK-2 cells. Moreover, BPP treatment decreased the expression levels of several transcriptional factors involved in regulating E-cadherin expression, including snail, twist and ZEB1. Additionally, the Hedgehog pathway was found to be closely correlated with renal fibrosis and EMT. Altogether, our results clearly demonstrated that BPP treatment effectively inhibited the Hedgehog pathway both in renal tissues of UUO mice and TGF-ß1-treated HK-2 cells. Thus, BPPs ameliorated RIF and EMT in vivo and in vitro via suppressing Hedgehog signalling pathway.

Identifying rotational symmetry axes in Kikuchi patterns by reciprocal vectors.

Symmetry analysis of the Kikuchi pattern is helpful to determine the crystal structure, and can significantly reduce the screening range of phase identification, thereby improving the accuracy and reliability of phase identification in electron backscatter diffraction (EBSD). Accurately identifying the symmetry axis from the Kikuchi pattern is the primary task of symmetry analysis. In this study, a new method was proposed to identify symmetry axes in Kikuchi patterns with the aid of reciprocal vectors. Taking the Kikuchi patterns of single-crystal silicon as a typical example, a method for drawing reciprocal vectors after strict projection correction is introduced. The complex task of identifying the symmetry axis is transformed into an intuitive judgment of the geometric relationship between reciprocal vectors, thus greatly simplifying the process. This method successfully elucidated information on six Kikuchi poles in three single-crystal silicon Kikuchi patterns, including 3-fold axes, 4-fold axes and asymmetric axes. The method can also distinguish between a 3-fold axis and an analogous 3-fold axis despite their only slight differences.

Symmetry analysis of the Kikuchi pattern is helpful to determine the crystal structure, and can significantly reduce the screening range of phase identification, thereby improving the accuracy and reliability of phase identification in electron backscatter diffraction (EBSD). Accurately identifying the symmetry axis from the Kikuchi pattern is the primary task of symmetry analysis. In our study, a new method was proposed to identify symmetry axes in Kikuchi patterns with the aid of reciprocal vectors. Taking the Kikuchi patterns of single-crystal silicon as a typical example, a method for drawing reciprocal vectors after strict projection correction is introduced. The complex task of identifying the symmetry axis is transformed into an intuitive judgment of the geometric relationship between reciprocal vectors, thus greatly simplifying the process. This method successfully elucidated information on six Kikuchi poles in three single-crystal silicon Kikuchi patterns, including 3-fold axes, 4-fold axes and asymmetric axes. The method can also distinguish between a 3-fold axis and an analogous 3-fold axis despite their only slight differences. It is indicated that the reciprocal vectors after projection correction can reflect the symmetry information well, making the identification of the symmetry axis more intuitive, which is helpful for symmetry analysis of the Kikuchi pattern, and lays an important foundation for phase identification using crystal symmetry.

Method for acquiring accurate coordinates of the source point in electron backscatter diffraction.

An electron backscatter diffraction device is an important accessory for a scanning electron microscope and can provide crystal structure orientation and phase content data through analysis of electron backscatter diffraction patterns. The acquisition of these data depends on pattern indexing, including interplanar angle calculation and crystal plane indexation. The coordinates of the source point are key points for interplanar angle calculation, and they vary with the movement of the incident beam. In this study, we first combined the grey gradient calculation with screen moving method to achieve accurate positioning of source point and obtained coordinates of source point with sub-pixel precision. The errors of three coordinates were 0.07%, 0.06% and 0.04%, respectively. By using this coordinate of source point to conduct interplanar angle calculation the maximum error was 0.53°, which was a good proof of the accuracy of source point positioning. Then we established the relationship between source point coordinates variation and incident beam movement. Coordinates can be given out based on the displacement of beam directly. And to illustrate the accuracy, interplanar angle calculation was performed and the maximum error was 0.81°. This means that the relationship between variation of source point coordinates and beam movement is highly accurate.

Moisture-sensitive metal-organic framework constructed from cobalt and 4-(4-pyridyl) benzoic acid for dispersive solid-phase extraction of polycyclic aromatic hydrocarbons in apple.

A moisture-sensitive metal-organic framework CoII (pybz)2 ·2DMF was synthesized and applied as the adsorbent of dispersive solid-phase extraction. The structure changed after water treatment due to the fact that two chelate carboxylate groups on the skeleton were transformed to monodentate because of the coordination of water molecules. The material showed good adsorption for fluorene, phenanthrene, fluoranthene, and pyrene in water because of the π-π conjugation and π-complexation effects. Coupled with high-performance liquid chromatography, a dispersive solid-phase extraction method of determining the content of fluorene, phenanthrene, fluoranthene, and pyrene in apple samples was established after optimizing the extraction conditions. Methanol containing 4% acetic acid was used as the effective eluent. The linearities were 0.5-1000 µg/kg for fluorene, phenanthrene and 5-1000 µg/kg for fluoranthene, pyrene. The limits of detection were 0.06-0.6 µg/kg, and the recoveries were 94.4-116.4%. The method has a high sensitivity for the determination of four polycyclic aromatic hydrocarbons in apple samples.