RNA-seq analysis of small intestine transcriptional changes induced by starvation stress in piglets.

Piglets may experience a variety of stress injuries, but the molecular regulatory mechanisms underlying these injuries are not well understood. In this study, we analysed the ileum of Large White (LW) and Mashen (MS) piglets at different times of starvation using chemical staining and transcriptome analysis. The intestinal barrier of piglets was damaged after starvation stress, but the intestinal antistress ability of MS piglets was stronger than LW piglets. A total of 8021 differentially expressed genes (DEGs) were identified in two breeds. Interestingly, the immune capacity (CHUK, TLR3) of MS piglets increased significantly after short-term starvation stress, while energy metabolism (NAGS, PLA2G12B, AGCG8) was predominant in LW piglets. After long-term starvation stress, the level of energy metabolism (PLIN5, PLA2G12B) was significantly increased in MS piglets. The expression of immune (HLA-DQB1, IGHG4, COL3A1, CD28, LAT) and disease (HSPA1B, MINPPI, ADH1C, GAL3ST1) related genes were significantly increased in two breeds of piglets. These results suggest that short-term stress mainly enhances immunity and energy metabolism in piglets, while long-term starvation produces greater stress on piglets, making it difficult for them to compensate for the damage to their bodies through self-regulation. This information can help improve the stress resistance of piglets through molecular breeding.

Targeting intestinal flora and its metabolism to explore the laxative effects of rhubarb.

Rhubarb, a traditional herb, has been used in clinical practice for hundreds of years to cure constipation, but its mechanism is still not clear enough. Currently, growing evidence suggests that intestinal flora might be a potential target for the treatment of constipation. Thus, the aim of this study was to clarify the laxative effect of rhubarb via systematically analyzing the metagenome and metabolome of the gut microbiota. In this study, the laxative effects of rhubarb were investigated by loperamide-induced constipation in rats. The gut microbiota was determined by high-throughput sequencing of 16S rRNA gene. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry was used for fecal metabolomics analysis. The data showed that rhubarb could significantly shorten gastrointestinal transit time, increase fecal water content and defecation frequency, improve gastrointestinal hormone disruption, and protect the colon mucus layer. Analysis of 16S rRNA gene sequencing indicated that rhubarb could improve the disorder of intestinal microbiota in constipated rats. For example, beneficial bacteria such as Ligilactobacillus, Limosilalactobacillus, and Prevotellaceae UCG-001 were remarkably increased, and pathogens such as Escherichia-Shigella were significantly decreased after rhubarb treatment. Additionally, the fecal metabolic profiles of constipated rats were improved by rhubarb. After rhubarb treatment, metabolites such as chenodeoxycholic acid, cholic acid, prostaglandin F2α, and α-linolenic acid were markedly increased in constipation rats; in contrast, the metabolites such as lithocholic acid, calcidiol, and 10-hydroxystearic acid were notably reduced in constipation rats. Moreover, correlation analysis indicated a close relationship between intestinal flora, fecal metabolites, and biochemical indices associated with constipation. In conclusion, the amelioration of rhubarb in constipation might modulate the intestinal microflora and its metabolism. Moreover, the application of fecal metabolomics could provide a new strategy to uncover the mechanism of herbal medicines.Key points• Rhubarb could significantly improve gut microbiota disorder in constipation rats.• Rhubarb could markedly modulate the fecal metabolite profile of constipated rats.

Comparative pharmacokinetics of the main active components in normal and ulcerative colitis rats after oral administration of Zingiberis Rhizoma-Ginseng Radix et Rhizoma herb pair and its single herb extracts by LC-MS/MS.

Zingiberis Rhizoma and Ginseng Radix et Rhizoma are usually used together for the treatment of ulcerative colitis in clinical practices. However, their compatibility mechanism remains unclear. In this study, a rapid and sensitive liquid chromatography with tandem mass spectrometry method was developed for simultaneous quantification of ginsenoside Re, ginsenoside Rg1, ginsenoside Rb1, and 6-gingerol in rat plasma after oral administration of Zingiberis Rhizoma-Ginseng Radix et Rhizoma herb pair and its single herb extracts. The calibration curves exhibited good linearity, with correlation coefficients of more than 0.993. The precision deviations of intra- and interday analysis were within 10.66%, and accuracy error ranged from -12.74 to 11.56%. The average recoveries of analytes were higher than 76.60% and the matrix effects were minimal. Thus, the validated method was successfully applied to a pharmacokinetic study of four ingredients in normal and ulcerative colitis rat plasma. The results indicated that the pharmacokinetic parameters of four analytes in normal and model groups showed significant differences. The larger exposure (the mean AUC0-t of ginsenoside Re, ginsenoside Rg1, ginsenoside Rb1, and 6-gingerol were increased by 50.93, 141.90, 3.68, and 37.25%, respectively) and slower elimination (the CLz/F of ginsenoside Re, ginsenoside Rg1, and 6-gingerol were decreased by 52.94, 83.64, and 32.18%, respectively) were observed in ulcerative colitis rats. Furthermore, compared with single herbs, the analytes in rat plasma after oral administration of combined extracts presented relatively high systemic exposure levels with AUC0-t > 2000 h·ng/mL and Cmax > 200 ng/mL. Collectively, the differences of pharmacokinetic characteristics revealed the synergistic effect of Zingiberis Rhizoma-Ginseng Radix et Rhizoma herb pair, which provided a valuable and reliable basis for its clinical application in the treatment of ulcerative colitis.

MicroRNA-212 promotes the recovery function and vascular regeneration of endothelial progenitor cells in mice with ischemic stroke through inactivation of the notch signaling pathway via downregulating MMP9 expression.

Ischemic stroke is a refractory disease caused by cerebral ischemic injury, which results in brain dysfunction. This study intends to investigate the effects of microRNA-212 (miR-212) on the recovery function and vascular regeneration of endothelial progenitor cells (EPCs) by inactivation of the Notch signaling pathway by binding to matrix metallopeptidase 9 (MMP9) in mice with ischemic stroke. According to the results of database retrieval systems and data analysis, MMP9 was predicted as a gene related to ischemic stroke and miR-212 is a potential regulating mRNA of MMP9. All 72 healthy adult C57BL6 mice were selected for middle cerebral artery occlusion (MCAO) establishment. Cerebral infarction was observed under triphenyltetrazolium chloride staining. A series of inhibitors, activators, and siRNAs were introduced to the verified regulatory functions for miR-212 governing MMP9 in ischemic stroke. Cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and tube-forming ability by tubule formation test. Reverse transcription quantitative polymerase chain reaction and Western blot analysis were used to detect the expressions of miR-212, MMP9, Hes-1, and Notch-1. The corresponding results demonstrated that the area of cerebral infarction and the number of neuronal necrosis increased in the MCAO group in contrast to the sham group. Meanwhile, upregulation of miR-212 or downregulation of MMP9 decreases the expressions of MMP9, Hes-1 Notch-1, increases cell proliferation and tube-forming ability and improves the pathological conditions of EPCs. Our study suggests that miR-212 promotes recovery function and vascular regeneration of EPCs through negative regulation of the Notch signaling pathway via downregulating expression of MMP9, thus provides a clinical theoretical basis for ischemic stroke therapy.

MicroRNA-137 protects neurons against ischemia/reperfusion injury through regulation of the Notch signaling pathway.

MicroRNAs (miRNAs) have emerged as novel regulators in various pathological processes including ischemic stroke. However, the precise role of miRNAs in ischemic stroke remains largely unknown. In this study, we investigated the role of miR-137 in the regulation of neuronal ischemia/reperfusion injury with oxygen-glucose deprivation and reoxygenation (OGD/R), a model of global brain ischemia. The results showed that miR-137 was significantly downregulated in neurons subjected to OGD/R treatment: OGD/R-induced cell injury was markedly inhibited by miR-137 overexpression and exacerbated by miR-137 suppression. Moreover, Notch1 was predicted as a target gene of miR-137 and verified by dual-luciferase reporter assay, real-time quantitative polymerase chain reaction, and western blot analysis. Through targeting of Notch1, miR-137 regulated the Notch signaling pathway. The blockade of the Notch signaling pathway reversed the effect of miR-137 suppression, whereas overexpression of the Notch intracellular domain abrogated the effect of miR-137 overexpression on OGD/R-induced cell injury. Overall, our study suggests that miR-137 regulated the Notch signaling pathway by targeting Notch1 to protect neurons from OGD/R-induced cell injury, providing a novel insight into understanding the molecular basis of ischemia stroke and a potential therapeutic target.

Prevalence, Risk Factors, Outcomes, and Treatment of Obstructive Sleep Apnea in Patients with Cerebrovascular Disease: A Systematic Review.

BACKGROUND: Obstructive sleep apnea (OSA) is known to increase the risk of cerebrovascular disease (CVD), and patients with CVD have high incidence of OSA. The study aimed to systematically evaluate the prevalence of OSA in patients with CVD. MATERIALS AND METHODS: Medline, Embase, Science Citation Index, Wanfang, CNKI, and Wiley Online Library were thoroughly searched to identify relevant studies. Random-effects models were used to calculate the pooled rate estimates. Meta-regression and subgroup analysis were performed to explore potential sources of heterogeneity. RESULTS: Thirty-seven studies with 3242 patients were analyzed. The prevalence of OSA (apnea hypopnea index [AHI] >10) ranged from 34.5% to 92.3%, the random-effects pooled prevalence was 61.9%. Furthermore, the prevalence of sleep disordered breathing (SDB) with AHI greater than 5 was 70.4%, with AHI greater than 20 was 39.5%, and with AHI greater than 30 was 30.1%. Only 8.3% of the SDB was primarily central apnea. Seventeen studies reported risk factors for OSA, 6 of which used multivariate analyses to extract risk factors. In univariate meta-regression analysis, male had higher prevalence than female (P = .041). OSA was associated with increased length of hospitalization in 2 studies, and 1 long-term study reported severe sleep apnea was associated with poor functional outcome. Among the 5 studies on treatment, 3 indicated that early treatment with CPAP was effective; the remaining studies did not find benefit from CPAP treatment and reported the CPAP acceptance was poor. CONCLUSIONS: There is high prevalence of OSA in patients with CVD (61.9%). Therefore, accurate diagnosis and treatment to OSA is very important so as to prevent CVD.

Maternally Expressed Gene 3 (MEG3) Enhances PC12 Cell Hypoxia Injury by Targeting MiR-147.

BACKGROUND/AIMS: Cerebral ischemia often leads to breakdown of blood-brain barrier (BBB) and vasogenic edema. It remains to be established whether MEG3 is responsible for the hypoxic damage in neural cells. This study aimed to investigate the role of MEG3 in the hypoxia-induced injuries of PC12 cells. METHODS: The PC12 cells were seeded and cultured under hypoxia and normoxia culture conditions. The cell viability determined by trypan blue exclusion, apoptosis using propidium iodide (PI) and fluorescein isothiocynate (FITC)-conjugated Annexin V staining, cell-migration using a modified two-chamber migration assay with a pore size of 8 µM and invasion using 24-well Millicell Hanging Cell Culture inserts with 8 µM PET membranes. RESULTS: Cell viability, relative migration and relative invasion decreased significantly in PC12 cells injured due to hypoxia as compared to control cells. An increase in apoptosis was also observed. The expression of MEG3 was up-regulated in hypoxia-injured PC12 cells. MEG3 overexpression enhanced hypoxia injuries, while MEG3 suppression attenuated the injuries. Meanwhile, MEG3 negatively regulated miR-147 expression. In addition, we found that the expression of Sox2 was increased in PC12 cells after hypoxia and miR-147 negatively regulated Sox2 expression through targets its 3'-UTR. Interesting, Sox2 activated NF-κB pathway and Wnt/ß-catenin pathway in PC12 cells. CONCLUSION: Considering the observations in our study, we can conclude that MEG3 aggravated the hypoxial injury in PC12 cells by down-regulating miR-147 gene and miR-147 further negatively regulated Sox2 expression.

The Distribution, Drug Susceptibility, and Dynamic Trends of Pseudomonas aeruginosa Infection in a Tertiary Hospital in China During 2016‒2022.

Background: Drug-resistant Pseudomonas aeruginosa infections rapidly increased and contributed to life-threatening nosocomial infections; however, the distribution, species, drug susceptibility and dynamic trends of P. aeruginosa infection in China remained unclear. This study was conducted to better understand the epidemiological data of increased P. aeruginosa infections from 2016 to 2022 in a hospital in China. Methods: This study involved 3301 patients infected with P. aeruginosa, diagnosed using a nosocomial infection surveillance system in a tertiary hospital between 2016 and 2022. The P. aeruginosa infections from 2016 to 2022 were assessed according to the hospital department and species, and the drug susceptibility was evaluated using 16 antimicrobial agents. Results: The P. aeruginosa infection prevalence in the hospital department was: Neurosurgery (14.30%), Emergency (13.30%), and Critical Care Medicine (11.69%). Samples for P. aeruginosa infection identification were from sputum (72.52%) and other secreta (9.91%). The P. aeruginosa infections demonstrated a greater sensitivity to amikacin (AMK, 91.82%), tobramycin (TOB, 82.79%), and gentamycin (GEN, 82.01%); however, P. aeruginosa infection demonstrated greater resistance to ticarcillin (22.57%), levofloxacin (21.63%), and ciprofloxacin (18.00%). Conclusion: The P. aeruginosa infections were commonly observed in the Neurosurgery, Emergency, and Critical Care Medicine departments and demonstrated greater sensitivity to AMK, TOB, and GEN than the other drugs.

Investigation of the material basis of Xiexin Tang to alleviate type 2 diabetes mellitus based on spectrum-effect analysis by UPLC-Q-TOF/MS.

Xiexin Tang (XXT) is a classic prescription for treating diabetes in clinical practices for thousands of years in China, which has been also proved by a large number of modern pharmacological studies. However, due to its complex composition, the bioactive ingredients of XXT is still unclear. In present researches, spectrum-effect relationship analysis is widely used to explore the material basis of traditional medical herbs, so this method was adopted in this study. Firstly, the extract of XXT was separated and enriched into 5 fractions by macroporous adsorption resin. Then, UPLC-Q-TOF/MS method was used for qualitative identification of components in each eluting part, and efficacy of each fraction was assessed by the T2DM rat model. Based on grey relational analysis and pearson bivariate correlation analysis, it was found that the components such as berberine, gallic acid, catechin, epicatechin, acteoside, berberastine and 1-O-galloyl-ß-D-glucose might be the main effective basis of XXT to improve T2DM.

Zingiber officinale and Panax ginseng ameliorate ulcerative colitis in mice via modulating gut microbiota and its metabolites.

Zingiber officinale and Panax ginseng, as well-known traditional Chinese medicines, have been used together to clinically treat ulcerative colitis with synergistic effects for thousands of years. However, their compatibility mechanism remains unclear. In this study, the shift of gut microbiome and fecal metabolic profiles were monitored by 16S rRNA sequencing technology and ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry analysis, respectively, which aimed to reveal the synergistic mechanism of Zingiber officinale and Panax ginseng on the amelioration of ulcerative colitis. The results showed that the relative abundance of beneficial bacteria (such as Muribaculaceae_norank, Lachnospiraceae NK4A136 group and Akkermansia) was significantly increased and the abundance of pathogenic bacteria (such as Bacteroides, Parabacteroides and Desulfovibrio) was markedly decreased after the intervention of Zingiber officinale-Panax ginseng herb pair. And a total of 16 differential metabolites related to ulcerative colitis were identified by the metabolomics analysis, which were majorly associated with the metabolic pathways, including arachidonic acid metabolism, tryptophan metabolism, and steroid biosynthesis. Based on these findings, it was suggested that the regulation of the gut microbiota-metabolite axis might be a potential target for the synergistic mechanism of Zingiber officinale-Panax ginseng herb pair in the treatment of ulcerative colitis. Furthermore, the integrated analysis of microbiome and metabolomics used in this study could also serve as a useful template for exploring the mechanism of other drugs.

The Bioavailability of Glycyrrhizinic Acid Was Enhanced by Probiotic Lactobacillus rhamnosus R0011 Supplementation in Liver Fibrosis Rats.

Glycyrrhizinic acid (GL) is clinically applied to treat liver injury, and the bioavailability of orally administered GL is closely related to the gut microbiota. Therefore, the dysbiosis of gut flora in liver injury could significantly influence GL bioavailability. Still, less is known about the impact of probiotic supplementation on the bio-absorption process of oral medication, especially under a pathological state. Herein, probiotic L. rhamnosus R0011 (R0011) with a high viability in the harsh gastrointestinal environment was selected, and the effect of R0011 on the GL bioavailability in rats was investigated. Four groups of rats (n = 6 per group) were included: the normal group (N group), the normal group supplemented with R0011 (NLGG group), CCl4-induced chronic liver injury model (M group), and the model group supplemented with R0011 (MLGG group). Our results showed that liver injury was successfully induced in the M and MLGG groups via an intraperitoneal injection of 50% (v/v) CCl4 solution. Healthy rats supplemented with R0011 could increase the bioavailability of GL by 1.4-fold compared with the normal group by plasma pharmacokinetic analysis. Moreover, the GL bioavailability of MLGG group was significantly increased by 4.5-fold compared with the model group. R0011 directly improved gut microbial glucuronidase and downregulated the host intestinal drug transporter gene expression of multidrug resistance protein 2 (MRP2). More critically, R0011 restored the gut microbiota composition and regulated the metabolic function, significantly enhancing the microbial tryptophan metabolic pathway compared with the pathological state, which may indirectly promote the bioavailability of GL. Overall, these data may provide possible strategies by which to address the low bioavailability of traditional medicine through probiotic intervention.

MicroRNA-410 inhibition of the TIMP2-dependent MAPK pathway confers neuroprotection against oxidative stress-induced apoptosis after ischemic stroke in mice.

Ischemic stroke (IS) is an acute cerebral event characterized by a high incidence rate, high disability rate as well as a high mortality. More recently, accumulative literature has provided evidence highlighting the role played by microRNAs (miRs) in the development of neurons. Hence, the aim of the present study was to investigate the neuroprotective role of miR-410 in IS. Microarray-based gene expression profiling of AMI was conducted in order to identify differentially expressed genes (DEGs) and the corresponding miRs regulating these genes. IS models were established to assess neurology on a scoring basis. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity and malondialdehyde (MDA) were all subsequently assessed. The functional role of miR-410 in IS was determined based on ectopic expression, knockdown and reporter assay experiments in hippocampal neurons. The expressions of microRNA-410, TIMP2, ERK, p38MAPK, JNK were all examined accordingly. The survival rate was assessed by MTT assay, and cell cycle and apoptosis by flow cytometry. After the loss of hippocampal neurons, infarct size as well as oxidative stress injury had been detected, microarray technology revealed that TIMP2 was differentially expressed in IS and that miR-410 regulated TIMP2. Initial observations revealed elevated levels of TIMP2 expression and MDA activity, in addition to evidence obtained indicated that the MAPK pathway had been activated along with decreased SOD, GSH-Px activity and miR-410 expression in IS mice. Ectopic expression of miR-410 was observed to inactivate the MAPK pathway, TIMP2 expression and hippocampal neuron apoptosis, while elevated hippocampal neuron survival rates and cell cycle entry were detected. Furthermore, TIMP2 as a direct target gene of miR-410, was determined to be negatively regulated by miR-410, while the MAPK pathway was found to be inhibited following TIMP2 knockdown. Our results revealed that the overexpression of miR-410 could ameliorate hippocampal neuron loss, reduce infarct size and oxidative stress injury in IS mice. Taken together, the key evidence of the current study elucidated the distinct nature of the inhibitory effect on IS as a result of overexpressed miR-410 whereby the conferral of neuroprotection was observed in oxidative stress-induced apoptosis post IS through the TIMP2-dependent repression of the MAPK pathway in mice.

Impact of acute and chronic stress hormone on male albino rat brain.

The present investigation aimed to evaluate the acute and chronic effect of stress (stress hormone) in male albino rat brain. Nor-epinephrine was used for the treatment and saline used for the control. Nor-epinephrine was dissolved in the saline and administered orally to the rats. Following nor-epinephrine administration, the brain was removed surgically at 6 h, 12 h and 45 days. Alanine tansaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) were significantly altered in the rats. Lipid peroxidation was measured as malondialdehyde (MDA), showed altered lipid peroxidation. Hematological markers such as packed cell volume (PCV), white blood cells (WBC), neutrophil, lymphocytes and hemoglobin were significantly altered compared to controls. Altered serum biochemical and hematological markers, lipid peroxidation and enzyme activities leads to adverse effect in the cellular metabolism and physiological activities of rats.

Primary application of PPE68 of Mycobacterium tuberculosis.

PPE68 protein is absent from BCG and the attenuated strains of Mycobacterium tuberculosis (MTB). In this study, the shuttle plasmid pBudCE4.1/PPE68/OriM was constructed and transformed into BCG to obtain PPE68 recombination BCG (PPE68-rBCG), and BALB/c mice were immunized with PPE68-rBCG to evaluate the immunological characterization of PPE68-rBCG. The level of lgG2a, IFN-γ, IL-12 and IL-4 in serum of immunized mice were detected, the proliferation response of spleen lymphocyte were measured, the frequency of CD4(+), CD8(+) and CD4(+)/CD8(+) were determined, and the spleen and lung tissue were prepared for pathological analysis. PPE68-rBCG was constructed successfully and could induce powerful Th1 immune response in mice. Besides, we took the purified recombination PPE68 (rPPE68) protein as diagnostic antigen to detect pulmonary tuberculosis patients (n=252) and extrapulmonary tuberculosis patients (n=66). We also used anti-PPE68 polyclonal antibody as coating antibody to detect specific antigen in the same serum samples. Our data provide an experimental basis for potential application of rPPE68 in the diagnosis of tuberculosis, especially for extrapulmonary tuberculosis.

Sorption characteristics and mechanisms of oxyanions and oxyhalides having different molecular properties on Mg/Al layered double hydroxide nanoparticles.

The sorption ability of fast-coprecipitated and hydrothermally-treated Mg/Al layered double hydroxide nanoparticles (FCHT-LDH) for various oxyhalides and oxyanions was evaluated. Interactions of oxyhalide such as monovalent bromate or oxyanions such as divalent chromate and divalent vanadate with FCHT-LDH were investigated using a combination of macroscopic (batch sorption/desorption studies and electrophoretic mobility (EM) measurements) and microscopic techniques (CHNS/O, XRD, FTIR, XPS, and EXAFS analyses). The sorption studies on various oxyanions and oxyhalides suggested that their sorption characteristics on FCHT-LDH were largely governed by their ionic potentials and molecular structures. Oxyanions which have ionic potentials higher than 7 nm(-1) were found to be more readily sorbed by FCHT-LDH than oxyhalides with ionic potentials lower than 5 nm(-1). The results obtained also demonstrated that trigonal pyramid oxyhalides showed a lower degree of specificity for FCHT-LDH than the tetrahedral coordinated oxyanions. From the macroscopic and microscopic studies, monovalent oxyhalide sorption on FCHT-LDH was postulated to occur mainly via anion exchange mechanism with subsequent formation of outer-sphere surface complexes. For polyvalent oxyanion sorption on FCHT-LDH, the mechanisms were possibly associated with both anion exchange and ligand exchange reactions, resulting in the coexistence of outer-sphere and inner-sphere surface complexes.