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This research aims to design and validate a machine learning model to predict the probability of urinary tract infections within 90 days post-urostomy in bladder cancer patients. Clinical and follow-up information from 317 patients who had urostomy procedures at the First Affiliated Hospital of Shanxi Medical University (May 2018-May 2024) were analyzed. The dataset were partitioned into training and testing sets, and feature selection was executed via the Least Absolute Shrinkage and Selection Operator regression technique. Seven machine learning algorithms were employed: Logistic Regression, K-Nearest Neighbors, LightGBM, Random Forest, XGBoost, Support Vector Machine, and Multi-Layer Perceptron. Performance metrics for the model were assessed using multiple evaluation indicators, including AUC, accuracy, sensitivity, specificity, Positive Predictive Value, Negative Predictive Value, and F1 score. SHapley Additive exPlanations and Local Interpretable Model-agnostic Explanations algorithms were applied for model interpretability. UTIs occurred in 22.08% of patients within 90 days after urostomy. The predictive model pinpointed eight important clinical features. Among the developed models, the SVM model demonstrated the best overall performance with AUC (0.835), accuracy (0.825), precision (0.583), recall (0.778), and F1 score (0.667). This model, designed to assess UTIs risk after urostomy in bladder cancer patients, has been deployed online for healthcare professionals at: https://zqmodel.shinyapps.io/shinydashboard_model/ ., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethical considerations: This study was approved by the Ethics Committee of the First Hospital of Shanxi Medical University (approval number NO.KYLL-2024–091) on April 11, 2024, which granted an exemption from obtaining informed consent due to the study’s nature and objectives., (© 2025. The Author(s).)

Polysaccharides from the dried tuber of Typhonium giganteum Engl. (TGEPs) were obtained by utilizing ultrasonic-assisted extraction (UAE) as the extraction method. The determination of optimal process parameters for the UAE of TGEPs (TGEP-U) was accomplished through the application of response surface methodology (RSM). The structural characteristics, antioxidant and hypoglycemic effects of TGEP-U and TGEPs obtained by hot water extraction (TGEP-H) were then compared. Consequently, the optimum extraction conditions predicted by RSM for TGEP-U were obtained as adding water at a ratio of 31 mL/g and extracted for 32 min under an ultrasound power of 440 W. In the verification experiment, the actual yield of TGEP-U was 7.32 ± 0.18 %. It was found that UAE could increase the yield and the total sugar content of TGEPs. Meanwhile, chemical composition analysis showed that both TGEP-U and TGEP-H were mainly composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, arabinose, and fucose, but the monosaccharide molar ratios were changed by UAE. Analysis of molecular weight (Mw) revealed the presence of three primary constituents within TGEP-U, and four main components in TGEP-H, and UAE reduced the average Mw of TGEPs. No obvious difference was found in the Fourier transform infrared spectroscopy analysis of TGEP-U and TGEP-H. The Congo red and Circular dichroism tests demonstrated that TGEP-U and TGEP-H had non-three helical structure. Scanning electron microscope observation further revealed that the aggregation of functional groups within TGEPs may be influenced by ultrasound, thereby affecting their powder morphology. TGEP-U has slightly poorer thermal stability than TGEP-H, which may be affected by ultrasonic cavitation effects. The results also indicated that TGEP-U had better antioxidant and hypoglycemic activity than TGEP-H. In summary, UAE is an effective method to extract and enhance the activity of TGEPs with enormous research value and potentials., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier B.V. All rights reserved.)

In this review, we comprehensively analyzed the distribution of microplastics （MPs） in major water ecosystems in China and the fate of MPs during the water treatment process. The removal efficiency of MPs with different colors, sizes, shapes, and materials was also discussed. The results showed that the abundance of microplastics in the aquatic environment was geographically variable and closely related to human activities. Fibrous and transparent （white） microplastics were the most common features in China's water ecosystems and water treatment plants, with polypropylene （PP）, polyethylene （PE）, and polystyrene （PS） being the most common polymer types of microplastics. The removal efficiency of MPs varied from different treatment processes significantly. Pre-treatment and primary treatment in wastewater treatment plants （WWTPs） contributed the most to the removal. In the secondary treatment, the sedimentation tank showed more efficiency than the biological treatment processes. Tertiary treatment processes demonstrated remarkable effectiveness in achieving terminal control of MPs, especially membrane technologies. On the contrary, aeration and hydrodynamic effects may have increased the abundance of MPs in WWTPs. In drinking water treatment plants （DWTPs）, coagulation-sedimentation processes were found to be the most effective in removing MPs, followed by filtration and disinfection processes. Further, both pre-treatment and post-treatment steps also made significant contributions to MPs removal.

Rheumatoid arthritis (RA) is a worldwide public health problem. Interventions to delay or prevent the onset of RA have attracted much attention in recent years, and researchers are now exploring various prevention strategies. At present, there is still no unified consensus for RA prevention, but targeting therapeutic windows and implementing interventions for at-risk individuals are extremely important. Due to the limited number of clinical trials on pharmacologic interventions, further studies are needed to explore and establish optimal intervention regimens and effective measures to prevent progression to RA. In this review, we introduce the RA disease process and risk factors, and present research on the use of both Western and Chinese medicine from clinical perspectives regarding RA prevention. Furthermore, we describe several complete and ongoing clinical studies on the use of Chinese herbal formulae for the prevention of RA., (© 2024. The Chinese Journal of Integrated Traditional and Western Medicine Press and Springer-Verlag GmbH Germany, part of Springer Nature.)

Polysaccharides from Pinelliae Rhizoma Praeparatum Cum Alumine (PPA) have various biological activities, but their properties after oral administration are not clear. In this study, the absorption, digestion and fermentation properties of PPA were studied using in vivo fluorescence tracking, in vitro simulated digestion and fecal fermentation experiments. The absorption experiment showed that fluorescence was only observed in the gastrointestinal system, indicating that PPA could not be absorbed. Simulated digestion results showed that there were no significant changes in the molecular weight, Fourier transform infrared spectroscopy (FT-IR) spectrum, monosaccharides and reducing sugar of PPA during the digestion process, showing that the overall structure of PPA was not damaged. However, the carbohydrate gel electrophoresis bands of PPA enzymatic hydrolysates after simulated digestion were significantly changed, indicating that simulated digestion might impact the configuration of PPA. In vitro fermentation showed that PPA could be degraded by microorganisms to produce short chain fatty acids, leading to a decrease in pH value. PPA can promote the proliferation of Bacteroideaceae, Megasphaera, Bacteroideaceae, and Bifidobacteriaceae, and inhibit the growth of Desulfobacteriota and Enterobacteriaceae. The results indicated that PPA could treat diseases by regulating gut microbiota, providing a scientific basis for the application and development of PPA., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Despite significant progress in diagnosis and treatment, asthma remains a serious public health challenge. The conventional therapeutic drugs for asthma often have side effects and unsatisfactory clinical efficacy. Therefore, it is very urgent to develop new drugs to overcome the shortcomings of conventional drugs. Natural polysaccharides provide enormous resources for the development of drugs or health products, and they are receiving a lot of attention from scientists around the world due to their safety, effective anti-inflammatory and immune regulatory properties. Increasing evidence shows that polysaccharides have favorable biological activities in the respiratory disease, including asthma. This review provides an overview of primary literature on the recent advances of polysaccharides from natural resources in the treatment of asthma. The mechanisms and practicability of polysaccharides, including polysaccharides from plants, fungus, bacteria, alga, animals and others are reviewed. Finally, the further research of polysaccharides in the treatment of asthma are discussed. This review can provide a basis for further study of polysaccharides in the treatment of asthma and provides guidance for the development and clinical application of novel asthma treatment drugs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Rheumatoid arthritis (RA) is a refractory autoimmune disease, affecting about 1% of the world's population. RA is divided into seronegative RA and seropositive RA. However, biomarkers for discriminating between seronegative and seropositive RA have not been reported. In this study, we profiled serum miRNAs in seronegative RA patients (N-RA), seropositive RA patients (P-RA) and healthy controls (HC) by small RNA sequencing. Results indicated that compared with HC group, there were one up-regulated and four downregulated miRNAs in N-RA group (fold change ≥ 2 and P value < 0.05); compared with P-RA group, there were two up-regulated and four downregulated miRNAs in N-RA group; compared with HC group, there were three up-regulated and four downregulated miRNAs in P-RA group. Among them, the level of hsa-miR-362-5p in N-RA group was up-regulated compared with that in HC group and P-RA group, and the level of hsa-miR-6855-5p and hsa-miR-187-3p in P-RA group was upregulated compared with that in N-RA group and HC group. Validation by qPCR confirmed that serum hsa-miR-362-5p level was elevated in N-RA group. Subsequently, by analyzing the target genes using RNAhybrid, PITA, Miranda and TargetScan and functions of differential miRNAs utilizing Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), we found that the target genes and molecular pathways regulated by miRNAs in seronegative RA and seropositive RA were roughly the same, and miRNAs in these two diseases may participate in the occurrence and development of diseases by regulating the immune system. In conclusion, this study revealed the profiles of serum miRNAs in seronegative and seropositive RA patients for the first time, providing potential biomarkers and targets for the diagnosis and treatment of seronegative and seropositive RA., Competing Interests: The authors declare that they have no competing interests., (© 2023 He et al.)

The pericarp of Zanthoxylum bungeanum maxim (PZM) is a commonly used spice and herbal medicine in China. In the present study, the structural characteristics of PPZM were investigated by saccharide mapping after enzymatic digestion by using high-performance thin layer chromatography (HPTLC) and polysaccharide analysis by using carbohydrate gel electrophoresis (PACE). The mechanisms of protective effects of PPZM on Aβ 25-35 -induced oxidative damage were explored in PC12 cells. The results showed that PPZM contained 1,4-α-D-galactosidic, 1,4-α-D-galactosiduronic, and (1→4)-β-D-glucosidic linkages. Pretreatment with PPZM significantly increased the cell viability of Aβ 25-35 -injured PC12 cells. Flow cytometry and Hoechst/PI staining indicated that PPZM gradually relieved the apoptosis of the Aβ 25-25 -treated cells. PPZM markedly decreased the ROS level of PC12 cells and suppressed Aβ 25-35 -induced oxidative stress by increasing the SOD level, and decreasing the level of MDA and LDH. The mRNA expressions of caspase-3 and Bax were significantly downregulated, and Bcl-2 expression was upregulated by treatment with PPZM. PPZM significantly increased the mRNA expression of Nrf2 and HO-1 in Aβ 25-35 treated cells. The results indicated that PPZM alleviated apoptosis and oxidative stress induced by Aβ 25-25 through the inhibition of mitochondrial dependent apoptosis and activation of Nrf2/HO-1 pathway. PPZM can be used as a potential protective agent against Aβ 25-25 -induced neurotoxicity.

Background: Glycine decarboxylase (GLDC), a key metabolic enzyme, participates in the regulation of the glycine metabolic pathway. Differential expression of GLDC is linked to the malignant growth of renal cell carcinoma (RCC) and may regulate tumor progression through other genes. However, the regulatory function of GLDC in RCC is currently unknown. The purpose of this work was to evaluate the roles of GLDC in the invasion, proliferation, and migration of RCC cells and elucidate the processes underlying RCC development., Methods: The expression of GLDC in RCC cell lines and tissues was identified by quantitative reverse transcription polymerase chain reaction (PCR) and western blot. A stably transfected cell line overexpressing GLDC was constructed using a lentiviral vector. Cell proliferation was detected using Cell Counting Kit-8 (CCK8) and EdU experiments, and scratch and transwell assays were used to determine migration and invasion capabilities. Furthermore, differential proteins were identified and obtained using high-performance liquid chromatography (HPLC)-tandem mass spectrometry (MS/MS) analysis. Finally, these differential proteins were analyzed by bioinformatics, including cluster analysis, subcellular localization, domain annotation, annotation of the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG), enrichment analysis, and study of protein-protein interactions., Results: GLDC expression was found to be lower in six RCC cell lines (786-O, A498, Caki-1, 769-P, OSRC-2, and ACHN) than in 293T cells and decreased in kidney cancer tissues compared to neighboring normal tissues. Overexpression of GLDC inhibited the proliferation of RCC cells as well as their migration and invasion abilities. Tandem mass tag analysis showed that 317 and 236 genes were downregulated and upregulated, respectively, when GLDC was overexpressed in A498 cells. Tandem mass tag analysis showed that 317 and 236 genes were downregulated and upregulated, respectively, when GLDC was overexpressed in A498 cells. Volcano plot showed these upregulated and downregulated proteins. Cluster analysis showed that differentially expressed protein screening can represent the effect of biological treatment on samples. Subcellular localization analysis showed differential proteins are mainly distributed in the nucleus, cytoplasm, mitochondria, plasma membrane, extracellular matrix, and lysosome. GO annotation showed many biological processes in the cells were changed, including "positive regulation of histone H3-K4 methylation", "cofactor binding", and "nuclear body". KEGG pathway analysis showed key pathways have all undergone considerable alterations, such as "cell cycle", "glyoxylate and dicarboxylate metabolism", and "threonine, glycine, and serine metabolism". Finally, highly aggregated proteins with the same or similar functions were acquired by analysis of the protein-protein interaction (PPI) network., Conclusions: These studies indicate that GLDC overexpression suppresses the invasion, proliferation, and migration of RCC cells and leads to the upregulation and downregulation of 236 and 317 genes, respectively., Competing Interests: The authors declare no conflict of interest., (Copyright 2022, Chen et al.)

Riboflavin is an essential micronutrient and a precursor of flavin mononucleotide and flavin adenine dinucleotide for maintaining cell homeostasis. Riboflavin deficiency (RD) induces cell apoptosis. Endoplasmic reticulum (ER) stress is considered to induce apoptosis, and C/EBP homologous protein (CHOP) is a key pathway involved in this process. However, whether RD-induced apoptosis is mediated by ER stress and the CHOP pathway remains unclear and needs further investigation. Therefore, the current study presents the effect of RD on ER stress and apoptosis in the human hepatoma cell line (HepG2). Firstly, cells were cultured in a RD medium (4.55 nM riboflavin) and a control (CON) medium (1005 nM riboflavin). We conducted an observation of cell microstructure characterization and determining apoptosis. Subsequently, 4-phenyl butyric acid (4-PBA), an ER stress inhibitor, was used in HepG2 cells to investigate the role of ER stress in RD-induced apoptosis. Finally, CHOP siRNA was transfected into HepG2 cells to validate whether RD triggered ER stress-mediated apoptosis by the CHOP pathway. The results show that RD inhibited cell proliferation and caused ER stress, as well as increased the expression of ER stress markers (CHOP, 78 kDa glucose-regulated protein, activating transcription factor 6) (p < 0.05). Furthermore, RD increased the cell apoptosis rate, enhanced the expression of proapoptotic markers (B-cell lymphoma 2-associated X, Caspase 3), and decreased the expression of the antiapoptotic marker (B-cell lymphoma 2) (p < 0.05). The 4-PBA treatment and CHOP knockdown markedly alleviated RD-induced cell apoptosis. These results demonstrate that RD induces cell apoptosis by triggering ER stress and the CHOP pathway.

The Huayu-Qiangshen-Tongbi (HQT) decoction, a Chinese medical formula, has been identified to show a potent therapeutic effect on rheumatoid arthritis (RA). However, the specific molecular mechanism of HQT in RA has not been well studied. In the present study, LPS-treated human rheumatoid fibroblast-like synoviocyte (FLS) MH7A cells and collagen-induced arthritis (CIA) mice were utilized as in vitro and in vivo models. Our results demonstrated that HQT could efficiently inhibit RA-induced inflammation by reducing the production of cytokines including tumor necrosis factor alpha (TNF- α ), interleukin-1 beta (IL-1 β ), and interleukin-6 (IL-6). Moreover, HQT significantly upregulated the expression of miR-125b. Besides, analysis of bioinformatics suggested casein kinase 2 (CK2) was a potential target of miR-125b. Luciferase reporter assay was performed and revealed that miR-125b suppressed CK2 expression in MH7A cells. Furthermore, miR-125b inhibited LPS-induced NF-kappa-B (NF- κ B) activation, which is a downstream target of CK2. In addition, the NF- κ B inhibitor ammonium pyrrolidinedithiocarbamate (PDTC) and NF-kappa-B inhibitor alpha (IkB- α ) enhanced the inhibitory effect of miR-125b on the expression of TNF- α , IL-1 β , and IL-6. Taken together, our study revealed that HQT could attenuate RA through upregulating miR-125b to suppress NF- κ B-induced inflammation by targeting CK2. The findings of this study should facilitate investigating the mechanism of HQT on RA and discovering novel therapeutic targets for RA., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Xiu-min Chen et al.)

Covalent organic framework nanospheres (COF NSs) have garnered special attention due to their uniform sphere morphology, adjustable particle size, and mesoporous microenvironment. However, methods to control an optimal particle size scale while achieving solution dispersibility and specific surface properties remain underdeveloped, which precludes many of the biomedical applications. Here, we propose and develop a general strategy to access simultaneous size control and surface functionalization of uniform spherical COF NSs in a single step using aspartic acid (d-/l-Asp) that plays center roles in an acid catalyst, hydrophilicity, size-controllable synthesis, and chiral enantiomer. In this study, for the first time, we have employed a surface chemistry engineering study to create a variety of nanoscale spherical COFs and subsequently measure parameters to evaluate the effectiveness of Asp in the regulation of the particle size. Moreover, the potential utilization of the d/l-enantiomeric Asp-COF NSs in preventing β-amyloid (Aβ) aggregation is investigated by analyzing their interactions with Aβ amyloids using a multitechnique experimental approach. To our knowledge, our strategy is the first synthesis of hydrophilic COF NSs with an optimal length scale and a chiral-selective targeting surface, which are crucial for the inhibition of Aβ fibrillation for Alzheimer's disease prevention.

Fusarium oxysporum f. sp. momordica was used as the target pathogenic fungus to screen actinomycetes that were isolated from rhizosphere soil of Momordica charantia by confrontation culture and antifungal tests of fermentation filtrate. The candidate strain 0250 had broad antifungal activity. According to cultural characteristics, physiological and biochemical properties, as well as average nucleotide identity analysis of the strains with similar homology, the strain 0250 was identified as Streptomyces rhizosphaericus . Its effects on growth promotion and control of bitter gourd wilt were evaluated in both greenhouse and field. The results showed that the plate inhibition percentage of S. rhizosphaericus strain 0250 against F. oxysporum f. sp. momordica was 69.2%, while the plate inhibition percentage against 17 plant pathogenic fungi reached 64.3%-85.6%. The suspension treatment of the strain could promote the growth and development of roots and stems and improve production of bitter gourd in pots and field. The control efficacy of Fusarium wilt of bitter gourd was 66.9% and 61.5%, respectively. When soils were treated with the strain 0250 suspension in advance and inoculated with the fungal pathogen, the inhibition percentage on the soil F. oxysporum reached 62.1%. The activity of phenylalanine ammonia-lyase, peroxidase and β-1,3-glucanase as well as root activity were significantly improved in bitter gourd seedlings. In summary, strain 0250 is an actinomycetes resource with biocontrol potential to Fusarium wilt of bitter gourd.

Talaromyces purpurogenus strain Q2 is a kind of beneficial microbe originally separated from the rhizosphere of healthy cucumber. In this study, we evaluated the biocontrol potential of strain Q2 against four soil-borne diseases by plate confrontation culture, potting in the greenhouse. We further estimated the control efficacy of strain Q2 combined metam-sodium fumigation against Fusarium wilt of bitter gourd in the field. The mechanism of strain Q2 controlling bitter gourd wilt and regulating soil microbial community was examined by plate dilution culture, high throughput sequencing and quantitative PCR. The results showed that strain Q2 could efficiently reduce disease incidence of Fusarium wilt of bitter gourd, potato stem canker, black shank of tobacco and black root rot of tobacco in the green house. Its biocontrol efficacy on black shank of tobacco and bitter gourd wilt was 75.3% and 63.4%, respectively. Biocontrol efficacy of strain Q2 on bitter gourd wilt was 51.0% in the artificial disease nursery inoculated pathogen of bitter gourd wilt, while the control efficacy of strain Q2 combined with soil fumigation technology was more than 80% in the same experiment condition. Strain Q2 application and soil fumigation altered soil microbial community composition and recovery trend. Metam-sodium fumigation significantly reduced the abundances of Fusarium oxysporum and causal agent of bitter gourd wilt. Strain Q2 further suppressed the efficient recovery trend of the pathogen. After application of strain Q2, Penicillium was enriched in soil, as well as the beneficial microbes involved in the suppression of F. oxysporum, such as Bacillus and Gaiella . Overall, after soil fumigation, biocontrol efficacy of strain Q2 on soil-borne diseases such as Fusarium wilt could attribute to the formation of beneficial microbial communities in soil and inhibition of strain Q2 on growth and development of F. oxysporum .

Chronic neuropathic pain poses a significant health problem worldwide, for which effective treatment is lacking. The current work aimed to investigate the potential analgesic effect of isoliquiritin, a flavonoid from Glycyrrhiza uralensis, against neuropathic pain and elucidate mechanisms. Male C57BL/6J mice were subjected to chronic constriction injury (CCI) by loose ligation of their sciatic nerves. Following CCI surgery, the neuropathic mice developed pain-like behaviors, as shown by thermal (heat) hyperalgesia in the Hargreaves test and tactile allodynia in the von Frey test. Repetitive treatment of CCI mice with isoliquiritin (p.o., twice per day for two weeks) ameliorated behavioral hyperalgesia to thermal (heat) stimuli and allodynia to tactile stimuli in a dose-dependent fashion (5, 15 and 45 mg/kg). The isoliquiritin-triggered analgesia seems serotonergically dependent, since its antihyperalgesic and antiallodynic actions were totally abolished by chemical depletion of spinal serotonin by p-chlorophenylalanine, whereas potentiated by 5-HTP (a precursor of 5-HT). Consistently, isoliquiritin-treated neuropathic mice showed escalated levels of spinal monoamines especially 5-HT, with depressed monoamine oxidase activity. Moreover, isoliquiritin-evoked antihyperalgesia and antiallodynia were preferentially counteracted by the 5-HT 1A receptor antagonist WAY-100635 delivered systematically or spinally. Of notable benefit, isoliquiritin was able to correct co-morbid behavioral symptoms of depression and anxiety evoked by neuropathic pain. Collectively, these findings demonstrate, for the first time, the therapeutic efficacy of isoliquiritin on neuropathic hypersensitivity, and this effect is dependent on the spinal serotonergic system and 5-HT 1A receptors., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Chronic neuropathic pain poses a significant health problem, for which effective therapy is lacking. The current work aimed to investigate the potential antinociceptive efficacy of isorhynchophylline, an oxindole alkaloid, against neuropathic pain and elucidate mechanisms. Male C57BL/6J mice were subjected to chronic constriction injury (CCI) by loose ligation of their sciatic nerves. Following CCI surgery, the neuropathic mice developed pain-like behaviors, as shown by thermal hyperalgesia in the Hargreaves test and tactile allodynia in the von Frey test. Repetitive treatment of CCI mice with isorhynchophylline (p.o., twice per day for two weeks) ameliorated behavioral hyperalgesia and allodynia in a dose-dependent fashion (5, 15, and 45 mg/kg). The isorhynchophylline-triggered antinociception seems serotonergically dependent, since its antinociceptive actions on neuropathic hyperalgesia and allodynia were totally abolished by chemical depletion of spinal serotonin by PCPA, whereas potentiated by 5-HTP (a precursor of 5-HT). Consistently, isorhynchophylline-treated neuropathic mice showed escalated levels of spinal monoamines especially 5-HT, with depressed monoamine oxidase activity. Moreover, the isorhynchophylline-evoked antinociception was preferentially counteracted by co-administration of 5-HT 1A receptor antagonist WAY-100635. In vitro , isorhynchophylline (0.1-10 nM) increased the Emax (stimulation of [ 35 S] GTPγS binding) of 8-OH-DPAT, a 5-HT 1A agonist. Of notable benefit, isorhynchophylline was able to correct co-morbidly behavioral symptoms of depression and anxiety evoked by neuropathic pain. Collectively, these findings confirm, for the first time, the disease-modifying efficacy of isorhynchophylline on neuropathic hypersensitivity, and this effect is dependent on spinal serotonergic system and 5-HT 1A receptors., (Copyright © 2020 Gao, Zhao, Wang, Yu, Wu and Zhao.)

A novel Heterotrophic Ammonia and Nitrate Bio-removal Over Nitrite (Hanbon) process, combining Short Nitrate Reduction (SNR) with Anaerobic Ammonia Oxidation (Anammox), was developed in a lab-scale continuous up-flow reactor. The substrate effects were investigated to characterize the performance of Hanbon process, and the corresponding microflora information was also revealed. Our results showed that the optimal substrate ratio of NH 4 + -N:NO 3 - -N:COD for the Hanbon process was 0.65:1:2.2. The volumetric nitrogen removal rate was up to 9.0 ± 0.1 kgN·m -3 ·d -1 at high influent substrate concentrations of NH 4 + -N 375 mg L -1 , NO 3 - -N 750 mg L -1 and COD 1875 mg L -1 , which was superior to the reported values of analogous processes. Moreover, the effluent total nitrogen concentration was able to meet the strict discharge standard (less than 10 mg L -1 ) at low influent substrate concentration of NH 4 + -N 26 mg L -1 , NO 3 - -N 40 mg·L -1 and COD 88 mg L -1 . Illumina-based 16S rRNA gene sequencing results showed that Halomonas campisalis and Candidatus Kuenenia stuttgartiensis were the dominant bacteria in the SNR section and Anammox section at high substrate concentration condition. However, Halomonas campaniensis and Candidatus Brocadia brasiliensis were raised significantly at low substrate concentration condition. Hanbon process provided in the present work was flexible of treating wastewater with various nitrogen concentrations, deserving further development., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Cucurbitaceae wilt of different hosts could be caused by Fusarium oxysporum. To clear the genetic diversity and phylogenetic relationship of F. oxysporum isolates from different Cucurbitaceae hosts and regions, genomic DNA of ninety-five strains of F. oxysporum isolated from different hosts and different regions were amplified by sequence-related amplified polymorphism (SRAP) molecular markers. All strains could be uniquely distinguished with 238 polymorphism bands which generated 100% of polymorphic ratio with 19 selective primer pairs. On average, each primer pairs amplified 12.5 loci and 12.5 polymorphic loci. A total of 166 bands of F. oxysporum f. sp. momordicae (FOM) were amplified, among which 145 polymorphic bands generated 87.4% of polymorphic ratio. On average, each primer combination amplified 8.7 loci and 7.7 polymorphic loci. This indicated that genetic variation of F. oxysporum was extensive. The genetic similarity coefficient of F. oxysporum was 0.68-0.99, and the average indexes of Nei's genetic diversity and Shannon were 0.2390 and 0.3718, respectively. Ninety-five strains of F. oxysporum were divided into four formae speciales from bitter gourd, cucumber, watermelon and melon, when the genetic similarity coefficient was 0.74. In SRAP dendrogram all the FOM isolates were gathered into one phylogenetic branch with the genetic similarity coefficient ranging from 0.78 to 0.99. The average indexes of Nei's genetic diversity and Shannon indices were 0.1811 and 0.2750, respectively. This indicated that genetic diversity of FOM was abundant, and the classification of phylogenetic group was related to geographic origin to some extent.

Decorin (DCN), a component of the extracellular matrix (ECM), participates in ECM assembly and influences cell proliferation and apoptosis in many mammalian tissues and cells. However, expression and function of DCN in the ovary remain unclear. This study cloned the full-length cDNA of goat DCN obtained from the ovary of an adult goat. Sequence analysis revealed that the putative DCN protein shared a highly conserved amino acid sequence with known mammalian homologs. The tissue distribution of DCN mRNA expression was evaluated by real-time PCR, and the results showed that DCN was widely expressed in the tissues of adult goat. Immunohistochemistry results suggested that DCN protein existed in the granulosa cells and oocytes from all types of follicles and theca cells of antral follicles. Moreover, hCG-induced DCN mRNA expression was significantly reduced by the inhibitors of protein kinase A, PI3K, or p38 kinase (P < 0.05), which are key mediators involved in hCG-induced DCN expression. Overexpression of DCN significantly increased apoptosis and blocked cell cycle progression in cultured granulosa cells (P < 0.05). Western blot analysis also showed that overexpression of DCN upregulated the expression levels of p21 protein (P < 0.05), whereas no effects were observed on the expression of Bax and Bcl-2 and on Bcl-2/Bax ratio (P > 0.05). These findings suggested that DCN regulates the apoptosis and cell cycle of granulosa cells., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Brain-derived neurotropic factor (BDNF) and its high-affinity receptor, tyrosine kinase receptor B, have been assumed to be involved in female reproduction and have recently shown to play an essential role in follicle activation and oocyte maturation. In this study, we analyzed the expression of miR-10b and BDNF in the ovary and discovered that the expression of miR-10b was higher in monotocous goat ovaries than in polytocous goat ovaries, whereas the expression pattern of BDNF in ovary was opposite. Moreover, human chorionic gonadotropin induced rapid and transient expression of BDNF messenger RNA and protein. In contrast, human chorionic gonadotropin upregulated miR-10b expression in a time-dependent manner. The BDNF gene was identified as a direct target of miR-10b using a dual-luciferase reporter assay. Transfection of granulosa cells with miR-10b decreased BDNF messenger RNA and protein levels. MiR-10b overexpression inhibited cell proliferation, whereas BDNF promoted cell proliferation. However, a combined treatment with miR-10b and BDNF promoted cell proliferation, indicating that the reintroduction of BDNF reversed the suppressive effect of miR-10b. These results demonstrate that miR-10b downregulates BDNF expression in granulosa cells by directly targeting the 3' untranslated regions and plays an important role in inhibiting granulosa cell proliferation by targeting BDNF., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Gray horses are born colored, and they then gradually lose their hair pigmentation. Tremendous progress has been made in identifying the genes responsible for graying with age in horses in recent years. Results show that gray coat color in horses is caused by a 4.6-kb duplication in intron 6 of the syntaxin 17 gene (STX17), which constitutes a cis-acting-regulatory mutation. However, little is known about the gray phenotype in native Chinese horses. This study was conducted to explore the frequency distribution of the gray mutation in native Chinese horse breeds. A total of 489 samples from 14 native Chinese horse breeds were genotyped for the STX17 duplication using a simplified conventional PCR-based method. The results show that the gray mutation was present in 12 native Chinese horse breeds, except the Balikun and Guanzhong breeds. The Chakouyi and Hequ breeds had the highest frequency of the gray mutation (P(G) = 0.367 and P(G) = 0.274, respectively). There was no significant geographical difference in the distribution of gray coat color across native Chinese horse breeds. Our results suggest that gray is a common coat color in Chinese horses.

Tissue inhibitor of metalloproteinase 1 (TIMP1) belongs to a group of endogenous inhibitors that control the activity of matrix metalloproteinases and other metalloproteinases. TIMP1 is ubiquitously expressed and implicated in many physiological and pathologic processes. In this study, the full-length complementary DNA of goat (Capra hircus) Timp1 was cloned from adult goat ovary for the first time to better understand the regulatory role of TIMP1. The putative TIMP1 protein shared a high amino acid sequence identity with other species. Real-time polymerase chain reaction results showed that Timp1 was widely expressed in adult goat tissues, and messenger RNA expression was higher in the ovary than in other tissues; meanwhile, increasing expression of Timp1 was also discovered during the process of follicle growth and corpus luteum. We then investigated Timp1 expression patterns in different types of ovarian follicular cells from goats. In small or large antral follicles, Timp1 expression was higher (P < 0.05) in theca cells than in granulosa cells, cumulus cells, and oocytes. Increasing expression of Timp1 in theca and granulosa cells was observed as the variation of the follicle size. Immunohistochemical analyses further revealed the presence of the TIMP1 proteins in follicles at all antral stages of development. The most intense staining for TIMP1 was observed in the theca cells and granulosa cells of large antral follicles and corpus luteum. Timp1 was highly (P < 0.05) induced in granulosa cells in vitro after treatment with the luteinizing hormone agonist, human chorionic gonadotropin. Treatments with forskolin, phorbol 12-myristate 13-acetate, or phorbol 12-myristate 13-acetate + forskolin could also stimulate Timp1 messenger RNA expression. The effects of human chorionic gonadotropin were reduced (P < 0.05) by the inhibitors of protein kinase A, protein kinase C, MAPK kinase, or p38 kinase, indicating that Timp1 expression could be adjusted by luteinizing hormone-initiated activation of these signaling mediators. Our results suggested that TIMP1 may be involved in regulating ovarian follicle development and ovulation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Microbial biocontrol agents (BCAs) are generally used for controlling plant diseases via antagonistic mechanisms including competition, antibiosis, parasitism, and cross-protection. Some BCAs can even promote plant growth, and provide induced systemic resistance (ISR), i. e., induce the plants to have resistance against pathogens including phytopathogenic fungi, bacteria and virus, and in some cases, pest insects and nematodes. ISR is characterized by non-specific, wide spectrum and systemic. It is phenotypically similar to the systemic acquired resistance (SAR) induced by the infection of pathogens, and with the same efficiency but without hypersensitive response (HR) and visible symptoms in plant as SAR, which is helpful to open a new way to develop and improve safer and environmentally friendly strategies for plant protection. In this paper, the research advances on ISR mediated by biocontrol fungi and bacteria, elicitors or determinants, and signaling transduction pathways were summarized, with more emphasis on the biochemical and cytological mechanisms of host plant defense reaction induced by free-living and endophytic BCAs. The potential application of ISR in biocontrol of plant diseases was also discussed.

A retinol-binding protein (RBP), synthesized and secreted by ovine allantois in vitro, was purified from culture medium. The protein consisted of three isoelectric variants (pI 5.3-6.1) of identical molecular masses of about 23,000 Da as determined by two-dimensional PAGE under reducing conditions. Thirty-one of the first 34 N-terminal amino acids of the purified protein were sequenced and shown to have complete homology with bovine placental and bovine plasma RBP. The ultraviolet absorption spectrum and fluorescence excitation and emission spectra of the purified ovine placental RBP indicated the presence of bound retinol. Metabolic labeling studies demonstrated that the protein was synthesized by placental membranes. Using antiserum to bovine placental RBP, ovine placental RBP was immunolocalized in trophectoderm of 13-day-old blastocysts and trophectodermal cells of the chorion, endodermal cells lining the allantois, and ectodermal cells lining the amnion of 23-, 45-, and 53-day-old conceptuses. Results from this study suggest that ovine placental membrane epithelia synthesize and secrete RBP. Transport, storage, and metabolism of retinol mediated by placental RBP may be essential for normal embryonic development during pregnancy.

An immunogold staining method was used in combination with epipolarization microscopic detection to demonstrate the presence of bovine placental retinol-binding protein in bovine extraembryonic membranes. Amnion, chorion and allantois were fixed in Bouin fixation fluid and embedded in polyethylene glycol 1500. Sections (5 mm) were cut and transferred onto Digene silanated slides and immunostained using rabbit antiserum raised against bovine placental retinol-binding protein followed by goat anti-rabbit IgG labeled with 1 nm gold. Gold particles after silver enhancement were viewed and photographed under epipolarization microscopy. Epithelial cells of all three membranes (i.e. amniotic ectoderm, chorionic trophectoderm, and allantoic endoderm) were immunoreactive, while mesodermal cells, collagen, and blood cells were not. These data, together with our previous observation that these three placental membranes synthesize and secrete retinol-binding protein, indicate that epithelial cells lining the amnion, chorion and allantois are the major sources of this protein. The presence of retinol-binding protein in placental membranes and their fluids may be indicative of an important role for retinol in placental differentiation and development.

Two high-affinity mAbs were prepared against Torpedo dystrophin, an electric organ protein that is closely similar to human dystrophin, the gene product of the Duchenne muscular dystrophy locus. The antibodies were used to localize dystrophin relative to acetylcholine receptors (AChR) in electric organ and in skeletal muscle, and to show identity between Torpedo dystrophin and the previously described 270/300-kD Torpedo postsynaptic protein. Dystrophin was found in both AChR-rich and AChR-poor regions of the innervated face of the electroplaque. Immunogold experiments showed that AChR and dystrophin were closely intermingled in the AChR domains. In contrast, dystrophin appeared to be absent from many or all AChR-rich domains of the rat neuromuscular junction and of AChR clusters in cultured muscle (Xenopus laevis). It was present, however, in the immediately surrounding membrane (deep regions of the junctional folds, membrane domains interdigitating with and surrounding AChR domains within clusters). These results suggest that dystrophin may have a role in organization of AChR in electric tissue. Dystrophin is not, however, an obligatory component of AChR domains in muscle and, at the neuromuscular junction, its roles may be more related to organization of the junctional folds.

Polyethylene glycol (PEG) is an excellent embedding medium for immunohistochemical studies. It provides structural preservation superior to frozen sections and increased sensitivity of antigen detection compared with paraffin sections. One limitation of PEG embedment is that PEG sections are difficult to handle and adhere poorly to glass slides. Here we present a simple and effective method for embedding tissues in PEG and transferring the resultant sections onto silanated glass slides. In addition, a method for silver enhanced colloidal gold immunostaining was combined with common dye staining to demonstrate the excellent structure preservation and sensitive antigen detection. Bovine chorionic membrane was fixed with Bouin's fixative, embedded in polyethylene glycol (PEG) 1500, cut into 5-microns sections, flattened over agarose blocks (10 x 10 x 2 mm3), and blotted onto Digene silanated slides. Slides were then washed in PBS, which removed the PEG and agarose blocks. Tissue sections were immunocytochemically stained with dilute antiserum raised in a rabbit against purified bovine placental retinol binding protein (bpRBP). Sections were washed and incubated with 1-nm colloidal gold-labeled goat anti-rabbit IgG. The immunogold particles were enhanced by silver staining (IGSS). Specimens were observed and photographed with an Olympus epipolarization microscope. The new method offered excellent morphological preservation of cell structure and the epipolarization microscopy provided high sensitivity for detection of specific immunogold-silver particles.

A procedure for fixing and immunostaining whole cells from primary cultures of ovine and bovine uterine gland fragments was used to identify keratin in intermediate filaments of epithelial cells to distinguish them from stromal cells. Colloidal gold encapsulated agarose-gelatin microbeads were coated with different proteins and used to investigate uptake by epithelial and stromal cells in culture. Microbeads were taken up by stromal cells and by epithelial cells on the outskirts of colonies. These cells formed ridges where they contacted and grew above stromal cells. Electron microscopy demonstrated that the microbeads had been internalized and appeared to be nontoxic. Individual cells could harbor more than 90 microbeads within their cytoplasm for at least seven to ten days with no apparent harm. Some cells with microbeads were seen to divide.

Agarose-gelatin microspherules about 0.5 micron or larger are prepared with emulsification of 4% agarose-gelatin sol containing 0.2 M N-octylglucoside in an organic phase composed of cyclohexane, egg lecithin, Span 80, and ethanol, followed by extraction of lipophilic components with cyclohexane and ether. Colloidal gold particles are then introduced into microspherules using gold chloride reacting at room temperature with tannic acid in a specified concentration range. After they have been coated with bovine serum albumin or mouse IgG, colloidal gold-labeled microspherules can be readily phagocytized by mouse L-cells and P388 cells after incubation for several hours. In addition to their use as a novel marker for phagocytosis, we discuss other potential uses for these colloidal gold-labeled microspherules.

The formation of acetylcholine receptor (AChR) clusters can be induced by basic polypeptide-coated latex beads in cultured Xenopus muscle cells. Here we investigated the development of acetylcholinesterase (AChE) at the bead-induced AChR clusters. AChE activity began to appear at the clusters after 1 day of bead-muscle coculture and was present at all of the bead-induced clusters within 4-7 days. Electron microscopy revealed that AChE reaction products were discretely localized within the cleft and the membrane invaginations at the bead-muscle contacts. Thus, the beads can mimic the nerve in inducing a local accumulation of both the AChRs and AChE, suggesting that the development of both specializations can be effected by a common stimulus.