Diosgenin Targets CaMKK2 to Alleviate Type II Diabetic Nephropathy through Improving Autophagy, Mitophagy and Mitochondrial Dynamics.

Diabetic nephropathy (DN) is a worldwide health problem with increasing incidence. Diosgenin (DIO) is a natural active ingredient extracted from Chinese yams (Rhizoma dioscoreae) with potential antioxidant, anti-inflammatory, and antidiabetic effects. However, the protective effect of DIO on DN is still unclear. The present study explored the mitigating effects and underlying mechanisms of DIO on DN in vivo and in vitro. In the current study, the DN rats were induced by a high-fat diet and streptozotocin and then treated with DIO and metformin (Mef, a positive control) for 8 weeks. The high-glucose (HG)-induced HK-2 cells were treated with DIO for 24 h. The results showed that DIO decreased blood glucose, biomarkers of renal damage, and renal pathological changes with an effect comparable to that of Mef, indicating that DIO is potential active substance to relieve DN. Thus, the protective mechanism of DIO on DN was further explored. Mechanistically, DIO improved autophagy and mitophagy via the regulation of the AMPK-mTOR and PINK1-MFN2-Parkin pathways, respectively. Knockdown of CaMKK2 abolished AMPK-mTOR and PINK1-MFN2-Parkin pathways-mediated autophagy and mitophagy. Mitophagy and mitochondrial dynamics are closely linked physiological processes. DIO also improved mitochondrial dynamics through inhibiting fission-associated proteins (DRP1 and p-DRP1) and increasing fusion proteins (MFN1/2 and OPA1). The effects were abolished by CaMKK2 and PINK1 knockdown. In conclusion, DIO ameliorated DN by enhancing autophagy and mitophagy and by improving mitochondrial dynamics in a CaMKK2-dependent manner. PINK1 and MFN2 are proteins that concurrently regulated mitophagy and mitochondrial dynamics.

Diosgenin Inhibits ROS Generation by Modulating NOX4 and Mitochondrial Respiratory Chain and Suppresses Apoptosis in Diabetic Nephropathy.

Diosgenin (DIO) is a dietary steroid sapogenin possessing multiple biological functions, such as the amelioration of diabetes. However, the remission effect of DIO on diabetic nephropathy (DN) underlying oxidative stress and cell apoptosis remains unclear. Here, the effect of DIO on ROS generation and its induced cell apoptosis was studied in vitro and in vivo. Renal proximal tubular epithelial (HK-2) cells were treated with DIO (1, 2, 4 µM) under high glucose (HG, 30 mM) conditions. DN rats were induced by a high-fat diet combined with streptozotocin, followed by administration of DIO for 8 weeks. Our data suggested that DIO relieved the decline of HK-2 cell viability and renal pathological damage in DN rats. DIO also relieved ROS (O2- and H2O2) production. Mechanistically, DIO inhibited the expression of NOX4 and restored mitochondrial respiratory chain (MRC) complex I-V expressions. Further, DIO inhibited mitochondrial apoptosis by ameliorating mitochondrial membrane potential (MtMP) and down-regulating the expressions of CytC, Apaf-1, caspase 3, and caspase 9, while up-regulating Bcl2 expression. Moreover, the ER stress and its associated cell apoptosis were inhibited through decreasing PERK, p-PERK, ATF4, IRE1, p-CHOP, and caspase 12 expressions. Collectively, DIO inhibited ROS production by modulating NOX4 and MRC complexes, which then suppressed apoptosis regulated by mitochondria and ER stress, thereby attenuating DN.

An overview of Fructus Meliae Toosendan: Botany, traditional uses, phytochemistry, pharmacology and toxicology.

Fructus Meliae Toosendan (FMT) is the dried and mature fruit of MeLia toosendan Sieb.et Zucc. It contains a variety of chemical constituents and reported to possess a variety of pharmacological activities. This review aims to provide a thorough and organized summary of botany, traditional uses, chemical ingredients, pharmacological actions, toxicity, quality control, and uses. In this review, we have compiled the data regarding FMT from 1994 to 2022 in the databases: Web of Science, PubMed, Google Scholar, CNKI, and Baidu Scholar. The keywords: "Fructus Meliae Toosendan", "botany", "traditional uses","chemical components", "pharmacological activity", "toxicity", "quality control" and "clinical application" have been used to collected the literature published in the online bibliographic databases. Based on the correlation of these documents and FMT, 126 articles were finally selected as references. This paper provides a reasonable summary of the 190 chemical components of FMT and its pharmacological effects and toxicity. Moreover, this paper also compiled the quality control studies and clinical applications. In the future, more experimental studies on FMT are needed to achieve the purpose of toxicity reducing and efficacy enhancing. This comprehensive review of FMT can provide a reference for subsequent relevant studies.

Cattle Encephalon Glycoside and Ignotin Ameliorate Palmitoylation of PSD-95 and Enhance Expression of Synaptic Proteins in the Frontal Cortex of a APPswe/PS1dE9 Mouse Model of Alzheimer's Disease.

BACKGROUND: Synaptic abnormalities in synaptic proteins are the initial hallmarks of Alzheimer's disease (AD). The higher level of palmitoylation of synaptic proteins was closely associated with amyloid-ß (Aß) in AD. Cattle encephalon glycoside and ignotin (CEGI) have been shown to act as multitarget neurotrophic agents in APPswe/PS1dE9 (APP/PS1) transgenic AD mice. However, it is not clear whether CEGI can influence Aß deposition or whether it does so by the regulation of protein palmitoylation and expression of synaptic proteins in transgenic AD mice. OBJECTIVE: In this study, we investigated the roles of CEGI in modulating postsynaptic density protein 95 (PSD-95) palmitoylation, Aß pathologies, and expression of synaptic-associated proteins in APP/PS1 mice. METHODS: Five-month-old APP/PS1 mice were treated intraperitoneally with 6.6 mL/kg of CEGI for 6 weeks. At the end of the treatment period, APP/PS1 mice were subjected to Morris water maze to test their cognitive functions. Acyl-biotinyl exchange (ABE) for PSD-95 palmitoylation, immunofluorescent staining for expression of PSD-95, N-methyl-D-aspartic acid receptor subunit 2B (NR2B), and synaptotagmin 1 (SYT1) were assessed in mouse brain sections. RESULTS: CEGI treatment in APP/PS1 mice significantly reduced Aß deposition, relieved memory deficits, and decreased PSD-95 palmitoylation while markedly increasing the expression of PSD-95, NR2B, and SYT1 in the frontal cortex. There was a significant correlation between Aß expression and PSD-95 palmitoylation in APP/PS1 mice. CONCLUSION: Our findings demonstrate that CEGI improved AD-like neuropathology, possibly by inhibiting PSD-95 palmitoylation, improving learning memory, and enhancing expression of synaptic-associated proteins, representing a potential therapy for AD treatment.

Inter-Eye Comparison in Highly Myopic Patients with Unilateral Myopic Traction Maculopathy.

PURPOSE: To evaluate the risk factors for myopic traction maculopathy (MTM) through inter-eye comparisons of asymmetric ocular features in patients with unilateral MTM. MATERIALS AND METHOD: Highly myopic patients with unilateral MTM were enrolled in the study. The results of comprehensive ophthalmologic examinations, color fundus images, and optical coherence tomography (OCT) were reviewed. MTM and myopic atrophic maculopathy was evaluated according to the ATN classification system. The status of the posterior vitreous detachment (PVD), posterior precortical vitreous pocket, vitreoretinal interface abnormalities, posterior staphyloma height (PSH), and the location of the protrusion of the sclera were assessed through OCT. RESULTS: Among the 54 eyes of 27 patients 48.89 ± 12.78 years of age, the affected eyes had worse best-corrected visual acuity, a longer axial length (AXL), greater PSH, and higher rates of posterior staphylomas, vitreovascular traction and epiretinal membranes (ERMs) than the fellow eyes (P < .001; P < .001; P < .001; P = .010; P = .002; P < .001). Thirteen cases present obvious anisometropia with an inter-eye AXL difference of more than 1 mm. In the 14 cases without obvious anisometropia, the eyes with MTM still had longer AXL, greater PSH and a higher ERMs rate than the fellow eyes (P = .039; P = .017; P = .001). Besides, in the 7 cases with an inter-eye AXL difference of less than 1 mm and asymmetrical stages of PVD, 5 cases with greater PVD in the affected eyes. Multiple risk factors coexisted in 96% of cases. CONCLUSIONS: In patients with unilateral MTM, a longer AXL, greater PSH, posterior staphyloma, vitreovascular traction, and ERMs were the main factors contributing to the occurrence of MTM. The process of PVD might involve in MTM development.

A Prospective Study of Intraocular Pressure Spike and Failure After Gonioscopy-Assisted Transluminal Trabeculotomy in Juvenile Open-Angle Glaucoma: A Prospective Study of GATT in JOAG.

PURPOSE: To identify risk factors for surgical failure after gonioscopy-assisted transluminal trabeculotomy (GATT) in juvenile open-angle glaucoma (JOAG). DESIGN: Prospective, interventional case series. METHODS: GATT was the initial surgery in 70 eyes of 70 patients with JOAG. Surgical success was defined as a postoperative intraocular pressure (IOP) of ≤21 mm Hg with at least a 20% reduction from preoperative IOP, with or without the use of antiglaucoma medication (qualified and complete success, respectively) at each postoperative visit. IOP spike was defined as IOP >30 mm Hg and an increase of at least 10 mm Hg from IOP before the spike, and then reduced to ≤21 mm Hg. RESULTS: The median age at the time of surgery was 19.3 years (range, 4.9-37.5 years) with a visual field mean deviation of -17.4 ± 10.6 dB. Mean IOP decreased from 31.3 ± 9.5 mm Hg preoperatively to 15.8 ± 2.7 at 12 months postoperatively. The complete and qualified success rates were 74.3% and 91.4%, respectively. An IOP spike occurred in 52 eyes (74%), with a median spike duration of 3.5 days (range, 1-21 days). Longer duration of IOP spike (P = .009) and older age at the time of surgery (P = .025) were both associated with worse surgical outcomes. Advanced disease was associated with prolonged IOP spike (P = .007). CONCLUSIONS: GATT provided excellent outcomes in patients with severe JOAG. Older age and longer duration of postoperative IOP spike are risk factors for failure. Severe cases are more likely to have longer durations of IOP spike. Frequent IOP monitoring during the early postoperative period is needed to detect IOP spikes in these patients.

Simultaneous Determination and Comprehensive Evaluation of Mineral Elements in Yellow Croaker Ear-Stone on Sale in China.

Yellow Croaker Ear-stone or Yunaoshi, is actually two kinds of fish otolith in China and has received increased attention in recent years as important folk medicine. For better understanding of this crude drug, a chaotic market circulation status investigation was carried out and seventeen samples with different varieties or producing areas were collected. In this study, pharmacodynamic components of nineteen varieties mineral elements of the seventeen samples were simultaneously determined by Inductively coupled plasma mass spectrometry (ICP-MS) method. The detected elements were categorized into the beneficial (Na, Mg, Ca, K, Fe, Mn, Zn, Sr, B) and unbeneficial elements (Cu, As, Cd, Hg, Al, Pb, Co, Ba, Cr and Ni) kinds and their concentrations were quantified. Then the principal component analysis (PCA) and hierarchical clustering analysis (HCA) were further applied to launch an exploratory analysis for Yunaoshi samples. The results showed that samples 1-3, 15-8, 15-3 ranked the top three from the perspective of beneficial elements and samples 1-3, 1-4, 15-2 ranked the top three based on the unbeneficial elements sides. Combined with HCA results, all samples can be used as the substitutes for Yunaoshi except for samples 1-3, 1-4 and 15-2 only judging from the perspective of mineral elements concentrations. In conclusion, simultaneous determination of mineral elements accompanied with PCA and HCA can not only provide pharmacogenetic reference for the medicinal material of Yunaoshi, but also establish a feasibility for exploring new crude resources or substitutes to this medicine.

High-Quality GaSe Single Crystal Grown by the Bridgman Method.

A high-quality GaSe single crystal was grown by the Bridgman method. The X-ray rocking curve for the studied GaSe sample is symmetric and the Full Width at Half Maximum (FWHM) is only 46 arcs, which is the smallest value ever reported for GaSe crystals. The IR-transmittance is about 66% in the range from 500 to 4000 cm-1. The photoluminescence spectrum at 9.2 K shows a symmetric and sharp excition peak in 2.1046 eV. The results indicate that the as-grown GaSe crystal is of high crystalline quality. The as-grown ε -GaSe crystal has a p-type conductance with the resistivity of 10³ Ω/cm, and the Hall mobility is ~25 cm² V-1 s-1. Few-layer GaSe crystals were prepared through mechanical exfoliation from this high-quality crystal sample. Few-layer GaSe-based photodetectors were fabricated, which exhibit an on/off ratio of 104, a field-effect differential mobility of 0.4 cm² V-1 s-1, and have a fast response time less than 60 ms under light illumination.

Existence results for a coupled system of nonlinear fractional multi-point boundary value problems at resonance.

By using the coincidence degree theory, we present an existence result for a coupled system of nonlinear fractional differential equations with multi-point boundary conditions at resonance.

Genetic variation in eight Chinese cattle breeds based on the analysis of microsatellite markers.

Genetic variability and genetic relationships were investigated among eight Chinese cattle breeds using 12 microsatellite markers. Three hundred and fifty-two alleles were detected and the average number of alleles per locus ranged from 8.33+/-1.67 in the Jiaxian breed to 21.33+/-5.60 in the Qinchuan breed with a mean value of 13.91. The total number of alleles per microsatellite ranged from 21 (INRA005, HEL1) to 40 (HEL13), with a mean of 29.33 per locus. The fixation indices at the 12 loci in the eight breeds were very low with a mean of 0.006. A principal components analysis and the construction of a neighbor joining tree showed that these eight Chinese cattle breeds cluster into three groups i.e. the Yanbian and Chinese Holstein, the Nanyang and Jiaxian, and the four remaining breeds.This clustering agrees with the origin and geographical distributions of these Chinese breeds.

Identification of microsatellites in cattle unigenes.

To identify EST-SSR molecular markers, 41,986 cattle UniGene sequences from NCBI were mined for analyzing SSRs. A total of 1,831 SSRs were identified from 1,666 ESTs, which represented an average density of 19.88 kb per SSR. The frequency of EST-SSRs was 4.0%. The dinucleotide repeat motif was the most abundant SSR, accounting for 54%, followed by 22%, 13%, 7% and 4%, respectively, for tri-, hexa-, penta- and tetra-nucleotide repeats. Depending upon the length of the repeat unit, the length of microsatellites varied from 14 to 86 bp. Among the di- and tri-nucleotide repeats, AC/TG (57%) and AGC (12%) were the most abundant type. Annotation of EST-SSRs was also carried out. Three hundred primer pairs were randomly designed using Prime Premier 5.0 program and Oligo 5.0 for further experimental validation.

Study on population genetic characteristics of Qinchuan cows using microsatellite markers.

To evaluate the genetic polymorphisms and to search for available molecular markers for Qinchuan cattle, 90 Qinchuan cows were genotyped with 12 microsatellite markers. A total of 247 alleles were detected, with the number of alleles ranging from 13 (INRA005) to 33 (HEL13), giving a mean number of 21 alleles per locus. The total and mean effective allele number were 142.6229 and 11.8852, respectively. Mean sampling variance of the allele frequency was 2.6036 x 10(-4). Allele size ranges of the 12 microsatellite loci were different. The observed heterozygosity and expected heterozygosity were from 0.7842 (INRA005) to 0.9775 (BM315) and 0.7952 (BM315) to 0.9446 (HEL13), respectively. Mean observed heterozygosity and mean expected heterozygosity were 0.9117 and 0.9047, respectively. Polymorphism information content values were from 0.7653 (INRA005) to 0.9420 (HEL13), and mean polymorphism information content of the 12 microsatellite loci was 0.8965. All the 12 microsatellite loci were highly polymorphic, which showed that there were rich genetic polymorphisms at these detected microsatellite loci in Qinchuan cows. At the 12 microsatellite loci, the mean fixation index was -0.0076, reflecting that the degree of heterozygote defect at these loci was not high and deviations from Hardy-Weinberg equilibrium were not significant.

Molecular cloning of bovine FABGL gene and its effects on bovine bioeconomic traits.

The complete CDS sequence of the bovine FABGL gene was determined by homology cloning approach combined with RT-PCR and 3'- and 5'-RACE. The results of sequence analysis and bioinformatics study showed that this cDNA contained 994 nucleotides, with a 780 bp open reading frame (ORF) flanked by a 16 bp 5'-UTR (incompletely) and a 198 bp 3'-UTR. The deduced amino acid sequence (260 AA) shows 88% identity with the corresponding sequence in humans. Two single nucleotide substitutions, one located in intron 5 (I5) at position 1 065 bp (Y = C/T) (GenBank: DQ409814) and the other in intron 8 (I8) at position 1 792 bp (R = A/G), were detected using the PCR-SSCP method. Analysis of the allele frequencies of the two polymorphic sites in three different cattle breeds (Angus, Hereford, and Simmental) with different genotypes showed large differences: in locus I8, cattle with the GG genotype showed higher beef performance index (BPI) (4.283 +/- 0.475 kg/cm) in comparison with cattle with the AA genotype (4.008 +/- 0.465 kg/cm) (P = 0.01). Regarding the ribeye area, cattle with the GG genotype showed significantly higher ribeye area (73.380 +/- 13.005 cm(2)) compared with cattle with the AA genotype (67.744 +/- 12.777 cm(2)) (p = 0.05). In locus I5, some associations for the average daily gain (ADG) were found at the significance level of 0.01 between three different genotypes (CC, CT, TT): cattle with the TT genotype showed the highest ADG (0.652 +/- 0.330 kg/d), whereas cattle with the CC genotype showed the lowest ADG value (0.421 +/- 0.178 kg/d).

[The effects of the GH, IGF-I and IGF-IBP3 gene on growth and development traits of Nanyang cattle in different growth period].

This study was conducted to identify polymorphisms of the Nangyang cattle's growth hormone (GH) and insulin-like growth factor-I (IGF-I) and insulin-like growth factor-I binding protein 3(IGF-IBP3) gene by the single nucleotide Polymorphisms and Polymerase chain reaction-based restriction fragment length polymorphism and to study association of polymorphisms identified in these genes with growth traits of the birth, 6 months, 12 months, 18 months, 24 months and 36 months. Results from the analysis showed a significant association of the BB genotype in the promoter of GH gene (GH-P5) with higher body length and body height during from 6 month to 18 month. From 24 month to 36 month, the IGF-IBP3 locus has a dominance modulation and control effect on backbody in Nanyang Cattle, and the BB genotype with higher Rump width than AA.

[Analysis on effect of microsatellite markers on milk production traits in Simmental].

12 microsatellite markers from 4 chromosomes were chosen to analyze the effect on 5 milk production traits (including fat, lactoprotein, lactose, dry matter content and somatic cell number) of 150 cows from a 6-male-line Simmental breeding core group. Results showed that the 12 microsatellite markers were highly polymorphic and their PIC (polymorphism information contents) were over 0.60, and the marker with the highest PIC (0.85) was ILST093. The heterozygosities were among 0.64 and 0.86. Effects of 12 microsatellite loci on milk production traits were analyzed. Results showed that ILST093 had a significant effect on somatic cell number (P<0.05). Effects of BMS711 on fat ratio and of BMS1905 on lactose both reached a significant level (P<0.05), BM6438 on milk production traits had no significant effect.

[Polymorphisms of insulin-like growth factor binding protein 3 gene and its associations with several carcass traits in Qinchuan cattle].

DNA samples from 60 Qinchuan cattle (Bos taurus) were analyzed with PCR-RFLPs and sequencing for insulin-like growth factor binding protein 3 (IGFBP3) gene. Fragments of 651 bp were amplified with two primers and the products of PCR were digested with restriction endonuclease HaeIII. The produced fragments showed three genotypes, namely AA,AB and BB after electrophoresis. Frequencies of the genotype AA,AB,BB and allele A,B were 0.7,0.28,0.02,and 0.84,0.16,respectively. Sequence analysis showed that a transversion of C-->A at 299 nt resulted in loss of the cleaved site of restriction endonuclease HaeIII and produced this polymorphism. This polymorphic locus of IGFBP3 gene was at Hardy-Weinberg equilibrium (P>0.05). The genotypes of AA,AB,BB slightly affected several slaughter and carcass traits of Qinchuan cattle. Dressing percentage,net meat percentage, striplion percentage, tenderloin percentage, ribeye percentage and tender shoulder percentage were decreased with the genotypes of AA,AB and BB in Qinchuan cattle, but it was not significant (P>0.05). Average ribeye area in individuals of AA genotype was significantly higher than that in individuals of BB genotype (P<0.05), and beef fat content in individuals of genotype AB and BB was significantly higher than that in individuals of AA genotype (P<0.01).