Relative Gene Expression Levels Research Articles

Sinomenine hydrochloride improves DSS-induced colitis in mice through inhibition of the Notch signaling pathway

ObjectiveTo study the therapeutic effect of sinomenine hydrochloride (SH) on dextran sodium sulfate (DSS)-induced colitis in mice as an animal model and the changes of Notch signaling pathway in colon tissue of mice after treatment.MethodsTwenty-four mice were randomly divided into control group, model group, SH low-dose group (20 mg/kg) and SH high-dose group (60 mg/kg), with 6 mice in each group. Disease activity index (DAI), colonic mucosal injury index and colonic histopathological score were calculated. The expression levels of related genes, proteins in Notch signaling pathway and inflammatory factors were quantified.ResultsSH can significantly reduce the symptoms of colitis mice, and can significantly reduce the DAI score (Model: 3.44 ± 0.27; SH-20: 2.50 ± 0.18; SH-60: 1.89 ± 0.17; P < 0.001) and histopathological injury degree (Model: 7.67 ± 0.52; SH-20: 5.17 ± 0.75, P < 0.01; SH-60: 3.33 ± 0.52, P < 0.001). SH can down-regulate the expression levels of Notch1, NICD1, Jagged1 and Hes1 proteins in colon tissue of colitis mice (Model: 1.92 ± 0.16, 1.83 ± 0.21, 2.23 ± 0.22, 1.91 ± 0.17; SH-20: 1.56 ± 0.12, 1.39 ± 0.13, 1.58 ± 0.12, 1.38 ± 0.11; SH-60: 1.24 ± 0.09, 1.23 ± 0.10, 1.23 ± 0.11, 1.22 ± 0.09; P < 0.01), and reduce the contents of serum pro-inflammatory cytokines TNF-α, IL-1β and IL-6 (Model: 718.53 ± 81.81, 51.62 ± 2.80, 444.07 ± 67.77; SH-20: 544.72 ± 90.03, 34.10 ± 2.90, 345.43 ± 43.40; SH-60: 434.11 ± 71.75, 29.44 ± 3.70, 236.11 ± 29.35; P < 0.001).ConclusionThe therapeutic effect of SH on DSS-induced colitis in mice may be related to inhibiting the overactivation of Notch signaling pathway.

Prmt1-mediated histone H4R3me2a methylation regulates the proliferation, migration and invasion of laryngeal cancer cells by affecting the expression level of NCOA5

BackgroundLaryngeal cancer is a common head and neck cancer, and its occurrence and development are closely related to a variety of epigenetic modifications. protein arginine methyltransferase 1 (PRMT1) is an important type I protein arginine methyltransferase, which catalyzes the monomethylation and asymmetric dimethylation of arginine and participates in the occurrence and development of a variety of cancers. Current research has found that the expression of PRMT1 is increased in laryngeal carcinoma tissues. Histone modifications play a key role in regulating gene expression and maintaining cellular function. In particular, histone H4 arginine 3 dimethylation (H4R3me2a) has been shown to be associated with the development of a variety of cancers. Nuclear Receptor Coactivator 5 (NCOA5) is an important nuclear receptor coactivator, which regulates gene expression through the interaction between various nuclear receptors and other transcription factors. The present study aimed to investigate how PRMT1-mediated H4R3me2a methylation affects the proliferation, migration and invasion of laryngeal cancer cells and to verify the role of NCOA5 in this process.MethodsThe expression of PRMT1 and NCOA5 was inhibited by siRNA mediated gene knockdown in laryngeal cancer cells. The changes of H4R3me2a protein levels were detected by Western Blotting, and cell proliferation, migration and invasion abilities were evaluated by CCK-8, cell scratch assay, Transwell migration and invasion assay. RT-qPCR was used to detect the mRNA expression levels of related genes. The overexpression experiment of NCOA5 was carried out by constructing overexpression vector to verify its effect.ResultsAfter PRMT1 knockdown, the expression of H4R3me2a in laryngeal cancer cells was significantly decreased, and the cell proliferation, migration and invasion abilities were weakened. Similarly, knockdown of NCOA5 expression also resulted in decreased H4R3me2a levels and attenuated cell proliferation, migration and invasion. Overexpression of NCOA5 partially restored H4R3me2a levels and cell proliferation, migration and invasion abilities.

Lentiviral Injection of Inter-α-Trypsin Inhibitor Heavy Chain 4 Promotes Female Spinal Cord Injury Mice Recuperation by Diminishing Peripheral and Central Inflammation.

Inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) acts as a mediator of inflammation and extracellular matrix stabilization. The current study intended to delve into the impact of ITIH4 on locomotor performance, nerve injury, neuroinflammation, systemic inflammation, and the downstream pathway in spinal cord injury (SCI) mice. Overexpression lentivirus of ITIH4 (LV-ITIH4) and negative control lentivirus (LV-NC) were intravenously injected into adult C57BL/6 mice on 7days before SCI surgery. All mice were euthanized on day 28 after SCI surgery, and their blood samples and spinal cord tissues were collected. Decreased relative gene expression and protein levels of ITIH4 were observed in SCI mice. LV-ITIH4 improved the locomotor performance compared to LV-NC in SCI mice. In spinal cord of SCI mice, LV-ITIH4 reduced apoptosis and increased survival of neurons compared to LV-NC. By comparison with LV-NC, LV-ITIH4 also reduced relative gene expressions of interleukin (IL)-6 and tumor necrosis factor-α in spinal cord of SCI mice. Moreover, LV-ITIH4 reduced microglia M1 polarization compared with LV-NC in spinal cord of SCI mice. In the serum, LV-ITIH4 decreased the protein levels of IL-6 and IL-1β compared to LV-NC in SCI mice. LV-ITIH4 also inhibited the nuclear factor kappa-B (NF-κB) pathway compared to LV-NC in spinal cord of SCI mice. ITIH4 enhances locomotor performance in SCI mice, and it inhibits nerve injury, neuroinflammation, systemic inflammation, and the NF-κB pathway in SCI mice.

DYRK2 controls GSTPI expression through ubiquitination and degradation of Twist1 to reduce chemotherapy resistance caused by EMT in breast cancer.

Breast cancer (BC) poses a significant global health challenge, with chemotherapy resistance, especially to docetaxel, remaining a major obstacle in effective treatment. The molecular mechanisms underlying this resistance are critical for developing targeted therapeutic strategies. This study aims to explore the role of dual-specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2), a member of the DYRK family, in docetaxel resistance in breast cancer cells and investigate its impact on cellular responses, including drug sensitivity and migration. Additionally, potential interactions between DYRK2 and Twist1, associated with epithelial-mesenchymal transition (EMT) and drug resistance, are explored. Docetaxel-resistant breast cancer cells were induced, and the expression levels of DYRK2, Twist1, and related genes were evaluated using real-time PCR and Western blotting. Lentivirus-mediated DYRK2 overexpression was employed to assess its effect on drug sensitivity, migratory ability, and Twist1 expression. The relationship between DYRK2 and Twist1 was examined, focusing on Twist1 ubiquitination. The impact of Twist1 on chemotherapy resistance and its binding to the Glutathione S-transferase Pi 1 (GSTP1) promoter were also investigated. Docetaxel-resistant cells exhibited down-regulated DYRK2 and up-regulated Twist1 expression. DYRK2 overexpression reversed drug resistance, decreased migration, and attenuated Twist1 and GST-π expression. DYRK2 was found to suppress Twist1 expression through ubiquitination, supported by decreased Twist1 phosphorylation and increased ubiquitination after DYRK2 overexpression. Twist1 overexpression counteracted DYRK2-induced drug sensitivity enhancement, promoting GST-π expression, EMT, migration, and proliferation. Twist1 was shown to bind to the GSTP1 promoter, enhancing its transcription. In vivo experiments confirmed DYRK2's ability to suppress chemoresistance in breast cancer cells. DYRK2 plays a pivotal role in overcoming docetaxel resistance in breast cancer cells by suppressing Twist1 expression through ubiquitination, impacting downstream signaling and cellular responses. This study provides valuable insights for developing targeted therapies to improve breast cancer treatment outcomes.

Identification and construction of prognostic clusters and risk-prognosis model based on aging-immune related genes in bladder cancer

BackgroundFaced with the current global ageing situation, advanced age has become a risk factor for bladder carcinogenesis progression and immunotherapy. Exploring the common mechanisms of aging and immune in bladder cancer and finding new prognostic markers and immunotherapeutic targets has become an urgent issue.MethodAging-immune related genes (AIGs) were collected from the public databases MSIGDB, HAGR and ImmPort, and hub AIGs were finally identified in the TCGA-BLCA disease cohort by expression, prognosis, and clinicopathological correlation analysis, and the correlation of hub AIGs with immune microenvironment, immunotherapeutic response, ferroptosis and m6A methylation was verified. Subsequently, prognostic clusters and risk-prognosis models for AIGs was constructed by cluster analysis and multifactorial Cox regression analysis, and the gene mutation and immune infiltration characteristics of the different clusters were explored. Finally, the expression level of related genes was verified by immunohistochemical experiments using patient samples from our medical center.Result145 potential prognostic AIGs were collected in bladder cancer and finally clarified NFKB1 and IL7 with significant expression differences, prognostic value and age correlation. By single gene analysis, hub AIGs were explored to be significantly correlated with immunotherapeutic response, immune microenvironment, ferroptosis and m6A methylation. Subsequently, the risk-prognosis model was constructed with Riskscore = (0.0581)*NFKB1 + (− 0.2285)*IL7. And prognostic clusters based on hub AIGs was performed by cluster analysis, which clarified that the high-risk group was the pro-cancer group, which had a lower mutation rate of hub genes and higher of neutrophil infiltration. Finally, immunohistochemistry of patients confirmed that IL7 and NFKB1 were underexpressed in bladder cancer, and the proliferation and migration ability of tumor cells were significantly decreased after overexpression of these genes.ConclusionThis study is the first to identify NFKB1 and IL7 as hub AIGs in bladder cancer, which provide new prognostic markers and immunotherapeutic targets.

Combination of Vitex pseudo-negundo methanolic-extract with cisplatin can induce antioxidant activity and apoptosis in HeLa and Caski cells.

Cisplatin-based chemotherapy as a common therapeutic regimen for cervical cancer patients, is becoming more and more ineffective due to high resistance. This urges the need for introducing novel metabolics such as botanical drugs with the capacity to increase the cisplatin effectiveness. In that regard, here we investigated the anticancer effects of the Cisplatin-Vitex pseudo-Negundo combination in cervical cancer cell lines. V. pseudo-Negundo fruits were dried and extracted methanolic fraction. The MTT assay was performed to evaluate cytotoxicity of both drugs in CaSki and HeLa cells. Then, apoptosis, ROS production, and cell cycling were assessed by flow cytometry assay in cells treated with V. pseudo-Negundo and Cisplatin and their combination. Also, the rate of cell migration and colony formation were measured, using wound healing and colony formation assay, respectively. Also, the expression level of related genes (CD133, BAX, BCL2, Casp-3/8/9, MMP-3) was evaluated using the RT-PCR method. The obtained results established that the V. pseudo-Negundo plant has medicinal properties to induce apoptotic and antioxidant signals. The combination treatment of methanol extraction and Cisplatin had a cytotoxic effect on cervical cancer cell lines (HeLa and CaSki) compared to monotherapy. Also, combination therapy resulted in an increased apoptosis rate and diminished ROS production in both CaSki and HeLa cell lines. Furthermore, V. pseudo-Negundo and Cisplatin combination therapy leads to cell cycle arrest in the G2-M and G0-G1 phase in HeLa and CaSki cell lines, respectively. Moreover, combination therapy decreased the colony formation and cell motility in both cell lines and upregulated caspases gene expression. The combination of V. pseudo-Negundo with Cisplatin therapy results in a significant anti-cancer and antioxidant effect compared to cisplatin, representing a promising candidate for future clinical investigations.

Physiological function of hydrophobin Hydph16 in cell wall formation in agaricomycete Pleurotus ostreatus

Hydrophobins are small-secreted proteins with both hydrophobic and hydrophilic regions, enabling the mycelium to break through the air-medium interface by reducing the medium surface tension. Over 20 putative hydrophobin-encoding genes have been predicted in the agaricomycete Pleurotus ostreatus. Three hydrophobin-encoding genes, vmh2, vmh3, and hydph16, were predominantly expressed in the vegetative mycelium. Despite these common properties, we have previously demonstrated the distinct functions of Vmh2 and Vmh3 in environmental stress resistance. In this study, we focused on hydph16 and found that Δhydph16 strains had sparser aerial mycelium than control strains. The cell wall thickness of Δhydph16 strains reduced by 40 % compared to that of control strains, but no significant differences were found in the relative chitin and glucan percentages or relative putative cell wall synthesis-related gene expression levels. Furthermore, unlike vmh2 and vmh3, hydph16 deletion did not change the hydrophobicity of the aerial mycelium. This study is the first to report that the lack of hydrophobin can lead to a significant change in aerial hyphae cell wall formation without altering the major cell wall polysaccharide composition. Additionally, this study revealed multiple roles for Hydph16, distinct from those of other highly expressed hydrophobins, Vmh2 and Vmh3. These results suggested that agaricomycetes, including P. ostreatus, have evolved to possess multiple hydrophobins with different functions.

Exogenous paclobutrazol effectively mitigates salt stress in ‘Duli’ (Pyrus betulifolia) seedlings by reducing Na+/K+ ratio, improving their osmotic substances, and activating the expression level of SOS and ABA pathway genes

Soil salinization is responsible for reductions in soil utilization by plants. Paclobutrazol (PAC) is an important plant growth regulator that is responsive to multiple biotic and abiotic stresses. In this study, we measured the impact of salt stress and PAC treatments on the growth, physiology, transcriptome, and related genes expression level of ‘Duli’ (Pyrus betulifolia). Our results show that NP treatment (200 mM NaCl + 10 μM PAC) improves this plant's morphological characters, decreases its Na+ content and Na+/K+ ratio from 9.33 % to 22.48 % compared to the NT (200 mM NaCl), increases its antioxidant enzyme activity, and inhibits its active oxygen as well as malondialdehyde content, and enables it to accumulate osmotic adjustment substances (i.e., proline and soluble sugar). NP treatment slows down chlorophyll degradation and regulation of endogenous hormones to safeguard photosynthesis. Further, combining the RNA–seq and q–PCR results reveals that NP increases the expression of salt overly sensitive (SOS) genes (SOS1, SOS2, SOS3), ion transporter (PbANN1, PbAKT), ABA receptor PbPYLs, and transcription factors (PbNAC, PbWRKY). In conclusion, this study's findings demonstrates that exogenous PAC is effective at countering salt stress–induced damage to ‘Duli’ seedlings, which can be applied to improve ‘Duli’ salt tolerance in pear production.

TNF-α Regulated Bidirectional Interaction Between Bone Marrow Mesenchymal Stem Cells and Articular Chondrocytes.

Articular chondrocytes (ACs) secrete a variety of extracellular matrix components to maintain the functions of articular cartilage. Degeneration of ACs leads to the degeneration of articular cartilage and consequently to osteoarthritis. The secretion of bone marrow mesenchymal stem cells (BMSCs) is capable of protecting ACs from degeneration, and thus BMSCs are widely applied to treat osteoarthritis. This study aims to explore whether BMSCs and ACs will affect the functions of each other through their secretions in the context of osteoarthritis. BMSCs and ACs isolated from rabbits were identified using flow cytometry and immunocytochemistry. Conditioned medium of BMSCs and ACs treated with 0, 5, 10, 20, and 40 ng/ml of tumor necrosis factor-alpha (TNF-α) were collected and used to treat ACs and BMSCs, respectively. The viabilities of ACs and BMSCs treated with condition medium were assessed using a Cell Count Kit-8 (CCK-8) kit. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), immunoblotting, and enzyme-linked immunosorbent assay (ELISA) methods were employed to evaluate the relative expression levels of genes and proteins, as well as the cytokine concentrations in the supernatant. Immunofluorescence and flow cytometry results indicated that the purity of isolated cells exceeded 95%. CCK-8 analysis showed that 6 hours of treatment with a conditioned medium did not affect the viability of BMSCs and ACs. However, treatment for 12 hours or longer significantly increased the viability of BMSCs (p < 0.05) and significantly decreased the viability of ACs (p < 0.01). RT-qPCR results demonstrated that the relative expression levels of Runx2 (1.15-3.91), Alp (1.06-2.84), TNF (BMSCs: 0.94-2.54; ACs: 1.03-2.64), IL6 (BMSCs: 0.98-2.78; ACs: 0.96-3.71), IL17A (BMSCs: 1.08-5.91; ACs: 0.90-4.20), and IL10 (BMSCs: 0.93-2.82; ACs: 0.89-2.25) genes in conditioned medium-treated BMSCs and ACs were dose-dependently elevated (p < 0.001) by TNF-α treatment. Immunoblotting analysis revealed that the expression levels of RUNX2 (0.53-0.86) and ALP (0.49-0.85) proteins were also dose-dependently elevated (p < 0.001) by TNF-α treatment. ELISA results showed similar TNF-α dose-dependent increases (p < 0.001) in the supernatant concentrations of pro-inflammatory cytokines TNF-α (BMSCs: 36.90 ± 0.75 to 199.38 pg/ml; ACs: 29.76 to 293.99 pg/ml), interleukin (IL)-6 (BMSCs: 4.96-48.24 pg/ml; ACs: 6.12-38.15 pg/ml), IL-17 (BMSCs: 3.06-28.99 pg/ml; ACs: 3.08-28.51 pg/ml), as well as the anti-inflammatory cytokine IL-10 (BMSCs: 6.34-65.02 pg/ml; ACs: 5.30-34.85 pg/ml). Together, these results indicate a TNF-α-regulated bidirectional interaction between BMSCs and ACs, deepening our understanding of the pathogenesis of osteoarthritis and aiding in its prevention and treatment.

Intercropping improves the yield by increasing nutrient metabolism capacity and crucial microbial abundance in root of Camellia oleifera in purple soil

Intercropping system influences the endophytic microbial abundance, hormone balance, nutrient metabolism and yield, but the molecular mechanism of yield advantage in Camellia oleifera intercropping with peanut is not clear. In this study, the C. oleifera monoculture (CK) and C. oleifera-peanut intercropping (CP) treatments in purple soil were conducted, and the physicochemical properties, gene expressions, signal pathways and crucial microbial abundances were investigated to reveal the molecular mechanism of the yield advantage of intercropped C. oleifera. The results showed that the intercropping system increased in contents of pigment, carbohydrate, available nitrogen and phosphorus in leaf and root, as well as the abundances of Burkholderia, Ralstonia, Delftia, Pseudoalteromonas and Caulobacter, enhanced the relative expression levels of CoSPS, CoGBE, CoGlgP, CoGBSS/GlgA genes to promote sugar metabolism, decreased the relative expression levels of CoASA, CoTSB, CoPAI, CoTDC and CoCYP71A13 genes for inhibiting IAA biosynthesis and signal transduction, as well as microbial diversity, Fusarium, Albifimbria and Coniosporium abundances in root, ultimately improved the fruit yield of C. oleifera. These findings indicate that intercropping system improves the fruit yield by enhancing the nutrient metabolism capability and crucial microbial abundances in root of C. oleifera in purple soil.

Metabolism Interaction Between Bacillus cereus SESY and Brassica napus Contributes to Enhance Host Selenium Absorption and Accumulation.

The use of beneficial bacteria to enhance selenium absorption in crops has been widely studied. However, it is unclear how the interaction between bacteria and plants affects selenium absorption in crops. Here, pot experiments and Murashige and Skoog medium (MS) experiments were performed. Transcriptomic analyses were used to reveal the interaction between Bacillus cereus SESY and Brassica napus. The results indicated that B. cereus SESY can significantly increase the biomass and selenium content of B. napus. The genes related to the colonization, IAA synthesis, and l-cysteine synthesis and metabolism of B. cereus SESY were significantly stimulated by B. napus through transcriptional regulation. Further verification results showed that l-cysteine increased selenium content in B. napus roots and shoots by 62.9% and 88.4%, respectively. B. cereus SESY and l-cysteine consistently regulated the relative expression level of genes involved in plant hormone, amino acid metabolism, selenium absorption, and Se enzymatic and nonenzymatic metabolic pathway of B. napus. These genes were significantly correlated with selenium content and biomass of B. napus (p < 0.05). Overall, IAA biosynthesis, and l-cysteine biosynthesis and metabolism in B. cereus SESY stimulated by interactions triggered molecular and metabolic responses of B. napus, underpinning host selenium absorption and accumulation.

Changes in Physiological Homeostasis in the Gills of Litopenaeus vannamei Under Carbonate Alkalinity Stress and Recovery Conditions

Carbonate alkalinity (CA) is the major toxic factor that interferes with the survival and growth of shrimp in saline–alkaline water. Gills are the main entry organ for CA toxicity in shrimp. In this study, low-salinity cultured Litopenaeus vannamei were exposed to 5 mmol/L CA stress for 7 days and then recovered for 7 days to explore the physiological changes in the gills under CA stress and recovery conditions at multiple biological levels. The results showed that CA stress increased the activities of antioxidative biochemical indexes (T-AOC, T-SOD, and POD) and the relative expression levels of romo1, nrf2, and gpx genes, while it decreased the relative expression levels of the sod and hsp70 genes. In addition, CA stress also increased the relative expression levels of genes involved in endoplasmic reticulum (ER) stress (bip, ire1, and xbp1), immunity (alf, crus, pen-3, lys, and propo), apoptosis (casp-3), detoxification metabolism (cyp450 and gst), and osmotic adjustment (ca, nka-α, nka-β, vatp, nhe, clc, aqp, tip4, and ccp). Although changes in some of the physiological indexes were reversed after the CA stress was relieved, they still could not effectively recover to the control level. These results reveal that CA stress has a negative impact on physiological homeostasis in the shrimp gills by inducing oxidation and ER stress and by interfering with immunity, apoptosis, detoxification, and osmotic adjustment.

Expression Levels of PF4, ALOX12, ITGA2B, F131A in Pregnant COVID-19 Survivors.

COVID-19 is viral illness caused by SARS-CoV-2. The immediate complications of COVID-19 are well defined and associated with increased mortality. A global effort is required to determine its effects on implantation, fetal growth and labor. Post COVID-19 recovery period presents a further challenge regarding service provision, prevention, and management. To assess the expression of Platelet Factor 4 (PF4), Arachidonate 12-lipoxygenase (ALOX 12), Integrin alpha-IIb (ITGA2B) & Coagulation Factor XIII A Chain F13A1 in post-acute COVID-19 survivors pregnant women. Prospective case control study, conducted on 400 pregnant women. Case group consists of 200 singleton pregnancies who had recovered from COVID-19 since 4-6 weeks before conception. Control group consists of 200 singleton pregnancies with no history for COVID-19. Expression levels of ALOX12, PF4, ITGA2B, and F13A1genes were determined using quantitative reverse transcription polymerase chain reaction method (qRT-PCR). Expression levels of ALOX12, ITGA2B, and F13A1, were significantly higher in the patients group (3.82±9.6, 6.63±8.45, and 8.9±9.1, respectively) (p < 0.05) compared to those in the control group (1.0±6.0, 1.0±8.1, and 0.6±7.6, respectively). No significant difference in PF4 expression between patients and control group (p = 0.3). Results obtained from enrichment analysis have also supported the above findings. Relative expression levels of these candidate genes could be distinguished between post-acute COVID-19 survivors' pregnant women and control group, significant relative gene expression of ALOX12, ITGA2B, and F13A1 may be associated with an increased risk of placenta-mediated adverse pregnancy outcomes.

Integrated physiological, energy metabolism, and metabonomic responses indicate the stress response in the hepatopancreas of Litopenaeus vannamei to nitrite stress

Nitrite is a toxic substance found in rearing water that affects shrimp health. The hepatopancreas is an important digestive, immune, and metabolic organ in the shrimp. In this study, shrimps (Litopenaeus vannamei) were separately exposed to 1 and 5 mg/L nitrite stress for 48 h, and the toxicity of nitrite in the hepatopancreas was explored by integrating histology, physiological indicators, energy metabolism, and metabolomics. Nitrite stress induced morphological changes and stress responses in the hepatopancreas. Specifically, physiology-related indices, such as the relative gene expression levels of antioxidants (ROMO1, Nrf2, GPx), endoplasmic reticulum stress (Bip, IRE1 and XBP1), and immune genes (ALF, Pen-3, Lys) were decreased, whereas the gene expression of apoptosis (Casp-3), detoxification (CYP450), and glutamic oxaloacetic transaminase (GOT) activity were increased. The activities of osmotic adjustment-related enzymes (NKA, CMA, and ATPase) also decreased. Energy metabolism-related indices, such as pyruvate and hepatic glycogen contents, increased, whereas glucose, lactic acid, triglyceride, and ATP contents and ATPase activity decreased, and the relative gene expression levels of carbohydrate metabolism (PDH, HK, and LDH) and electron-transport chain genes (CytC, COI and CCO) decreased, and the expressions of lipid metabolism (AMPK, SREBP, and FAS), tricarboxylic acid cycle (MDH, CS, IDH and FH) genes were also disturbed. The metabolic pattern of the hepatopancreas was affected by nitrite stress. Glycine, serine, and threonine metabolism were highly affected, and more functional amino acids varied in the 5 mg/L nitrite stress group. These results reveal the toxic effects of nitrite stress on the stress response, physiology, energy metabolism, and metabolite homeostasis in the hepatopancreas of shrimp. Several potential metabolite biomarker candidates were identified for toxicological evaluation.

Lactobacillus johnsonii CCFM1376 improves hypercholesterolemia in mice by regulating the composition of bile acids

Aim: Strains with high bile salt hydrolase (BSH) activity have the potential to regulate cholesterol metabolism. This study aimed to assess the alleviating effect of Lactobacillus johnsonii (L. johnsonii ) CCFM1376, a strain with high BSH activity, on mice with hypercholesterolemia and explore the mechanism of its effect through the modulation of bile acid metabolism. Methods: The BSH activity was measured using the ninhydrin method. C57BL/6J mice were given a high-cholesterol diet to induce hypercholesterolemia with simultaneous gavage of L. johnsonii CCFM1376 for 8 weeks. The biochemical parameters in the serum and liver of hypercholesterolemic mice were measured to assess the alleviating effect of L. johnsonii CCFM1376 on hypercholesterolemia. Bile acid content in the mouse liver, serum, distal ileum contents, and feces was determined using liquid chromatograph mass spectrometer (LC-MS). RNA was extracted from mouse ileum and liver, and the expression levels of relative genes implicated in bile acid metabolism were measured by quantitative real-time PCR (qPCR). Results: Compared to the model group, the group treated with L. johnsonii CCFM1376 exhibited significantly reduced levels of serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C), along with a significant increase in high density lipoproteins cholesterol (HDL-C) level. Moreover, hepatic levels of TC and LDL-C in the CCFM1376 group also decreased significantly. Furthermore, the content and amount of unconjugated bile acids in the hepatic-enteric circulation of the L. johnsonii CCFM1376 group significantly increased, and the total bile acid content in the feces also significantly increased. In the L. johnsonii CCFM1376 group, the relative expression levels of ileal farnesoid X receptor (FXR) and fibroblast growth factor 15 (FGF15) were downregulated, while the relative expression level of CYP7A1 was upregulated. Conclusion: These results indicated L. johnsonii CCFM1376 improves hypercholesterolemia in mice by regulating the composition of bile acids. This provides a reference for probiotic strategy to regulate cholesterol metabolism.

Limosilactobacillus Fermentun ZS09 Can Improve Antibiotic-Induced Motor Dysfunction in Mice by Regulating the Brain-Gut Functions.

When consumed in appropriate quantities, probiotics, which are live microorganisms, are good for health. In this study, a mouse model of antibiotic-induced dyskinesia was established using a sterile mixed antibiotic solution to investigate the preventive impact of Limosilactobacillus fermentum ZS09 (LFZS09) on this condition in mice. Following modeling, alterations in the serum and brain tissue of mice were assessed for motor measures such as running and swimming, malondialdehyde (MDA), superoxide dismutase, reduced glutathione, interleukin (IL-6, IL-10), and tumor necrosis factor (TNF)-α. The mouse cecum was used to evaluate the relative mRNA expression levels of the intestinal barrier genes, namely occludin-1, zona occludens-1 (ZO-1), and claudin-1. The relative mRNA expression levels of cAMP-response element binding protein (CREB), extracellular signal-regulated kinase (ERK), and brain-derived neurotrophic factor (BDNF) genes in the mouse brain tissue were also evaluated. Compared with the model group, LFZS09 considerably increased the swimming and running duration of mice, significantly decreased the levels of the inflammatory factors TNF-α and IL-6 and increased SOD expression in the mouse brain, and decreased MDA accumulation in the mouse brain and serum. Furthermore, LFZS09 upregulated occludin-1 gene expression in the cecal tissue to maintain the intestinal barrier, which in turn maintained the normal physiological function of the body. LFZS09 also enhanced the effect of BDNF and increased the expression of BDNF metabolic pathway-related genes, namely CREB, ERK1/2, and BDNF, in the mouse brain tissue. LFZS09 increased the number of Lactobacillus and Bifidobacterium in the gut of mice with motor dysregulation, and decreased the number of Enterococcus and Clostridium perfringens. The findings indicate that LFZS09 regulates antibiotic-induced motor impairment in mice, thereby offering a theoretical foundation for future studies and probiotic or parabiotics production aimed at augmenting motor function.

Exogenous 5-aminolevulinic acid promotes carotenoid accumulation in tomato fruits by regulating ethylene biosynthesis and signaling.

5-Aminolevulinic acid (ALA) can not only improve fruit yield and quality, but also increase the lycopene content in tomato fruits. Furthermore, ALA has been shown to promote system-2 ethylene production in tomato fruits. However, the specific interactions between ALA and ethylene during fruit ripening remain unclear. In this study, we treated tomato fruits with ALA, 1-aminocyclopropane-1-carboxylic acid (ACC), aminooxyacetic acid (AOA) + AgNO3, and AOA + AgNO3 + ALA and analyzed ethylene emissions, carotenoid contents, and the relative gene expression levels related to fruit ripening, carotenoid contents, ethylene synthesis, and signal transduction. The ALA treatment significantly enhanced ethylene bursts and carotenoid accumulation, and significantly upregulated the expression of ethylene and carotenoid-related genes, such as SlACS2, SlACS4, SlACO1, SlPSY1, and SlPDS. We also observed that the gene expression levels associated with carotenoid synthesis were downregulated in fruits treated with a combination of ethylene inhibitors (AOA + AgNO3). However, there was a significant upregulation in the gene expression levels associated with carotenoid synthesis and an increase in carotenoid content when fruits were treated with AOA + AgNO3 + ALA. After silencing SlACO1 expression, the total carotenoid content and SlPSY1 expression decreased significantly, while this effect was reversed after exogenous application of ALA. These results indicated that ALA promotes carotenoid accumulation in tomato fruits by promoting ethylene biosynthesis. In conclusion, our results highlighted the role of ALA in promoting carotenoid accumulation and ripening in tomato fruits by regulating ethylene synthesis, thereby providing a novel strategy for improving fruit quality.

Characterization of Cytoskeletal Profilin Genes in Plasticity Elongation of Mesocotyl and Coleoptile of Maize Under Diverse Abiotic Stresses.

The plasticity elongation of mesocotyl (MES) and coleoptile (COL) largely determines the morphology of maize seedlings under abiotic stresses. The profilin (PRF) proteins play a pivotal role in cytoskeleton dynamics and plant development via regulating actin polymerization. However, little is known about whether and how the expression of the ZmPRF gene family regulates MES and COL elongation in maize under adverse abiotic stresses. Here, a total of eight ZmPRF gene members were identified in the maize genome. They were mainly located in the cytoplasm, chloroplast, and mitochondrion, and clearly divided into four classes, based on phylogenetic analysis. Segmental duplication was the main driver for the expansion of ZmPRF genes. Ka/Ks analysis indicated that most ZmPRF genes were intensely purified and selected. Promoter cis-element analysis suggested their potential roles in response to growth and development, stress adaption, hormone response, and light response. The protein-protein interaction network and two independent RNA-sequencing analyses revealed that eight ZmPRF genes and their thirty-seven interacting genes showed varied expression patterns in MES and COL of three maize genotypes under different sowing depths, 24-epibrassinolide application, and light spectral-quality treatments, of which ZmPRF3.3 was a potential core conserved gene for breeding application. Moreover, the quantitative real-time PCR (qRT-PCR) verified that the relative expression levels of most ZmPRF genes in MES and COL under above treatments were significantly correlated with the plasticity elongation of MES and COL in maize. Therefore, these results perform a comprehensive overview of the ZmPRF family and will provide valuable information for the validation of the function of ZmPRF genes in maize development under diverse abiotic stress.

Chronic toxic effects of chloroxylenol exposure on Rana chensinensis: Insights from endochondral ossfication

Chloroxylenol (para‑chloro-meta-xylenol, PCMX), is a widely used antimicrobial agent and can remain in the aquatic environment. Although toxicity studies related to PCMX on the aquatic animals like zebrafish and Brachionus koreanus have been reported, there are few reports in the ecological risk of amphibians. In this study, the toxicity of different concentration (143, 14.3, 1.43 μg/L) of PCMX treatments on the endochondral ossification and body condition of Rana chensiensis tadpoles was investigated at environmentally relevant concentrations during metamorphosis. The chronic exposure of PCMX decreased bone length and ossification of limbs, caused changes of thyroid gland structure and ossification related gene expression levels. Besides, we found that R. chensiensis developed rheumatoid arthritis. Therefore, these results provided valuable evidence that the ecological risk of PCMX that will negatively affect the body condition, thyroid hormones homeostasis and skeletal development of R. chensiensis tadpoles.

Morpho-molecular evaluation for developmental competence of oocytes retrieved through transvaginal ovum pick-up from FSH-stimulated Tharparkar donor cows (Bos indicus).

The study was conducted on indigenous Tharparkar cow (Bos indicus) to evaluate FSH stimulation on follicular attributes, oocyte recovery and morpho-molecular developmental competence parameters concerning oocyte quality. A total of 20 OPU sessions were performed, which included 10 sessions in each FSH stimulated at the dose of 130 µg divided into four sub-doses and non-stimulated. Findings on the size of follicles having ≥6 mm showed a significantly higher, however an opposite trend was observed in the case of smaller sized follicle (<6 mm) between stimulated and non-stimulated respectively. The stimulated cows had a significantly higher number as well as the percentage of oocytes of Grade A, having a diameter ≥120 µm and BCB+VE as compared to the non-stimulated cows. The relative mRNA expression profile of GDF9, BMP15, PCNA and BCL-2 genes was higher and BAX was lower in the FSH-stimulated cow. These results indicated that FSH stimulation before OPU in Bos indicus cows has a significant impact on follicle size, oocyte yield, recovery, and their quality with respect to COC's, diameter and BCB+VE oocytes. Further, a significant increase in the relative mRNA expression levels of GDF9, BMP15 and PCNA genes in the FSH-stimulated group suggests that FSH plays a key role in modulating the expression of these important candidate genes and thus influencing oocyte quality. The higher mRNA expression of BCL-2 genes and concomitantly lower expression of BAX gene in FSH Stimulated cows indicates the protective role of these genes and preventing programmed cell death and thus promoting cell survival, quality and embryo development.