MOD Group Research Articles

Characteristics of Gut Microbiome in the Murine Model of Pancreatic Cancer with Damp-Heat Syndrome.

Murine models of pancreatic cancer with damp-heat syndrome were established based on two methods to explore the differences in the composition of intestinal flora and to seek characteristic genera with potential for model evaluation. In our study, thirty-four C57BL/6J male mice were randomly divided into a control group (Con), a model group (Mod), a classic damp-heat syndrome group (CDHS), and a climate-chamber group (CC). CDHS and CC groups were fed with a high-fat diet and glucose water, while the CDHS group was given 2.4 g/kg alcohol by gavage for 10 days, and the CC group was placed in a climatic chamber with a set temperature of (32 ± 1) °C and humidity of (92 ± 2)% for 10 days. The Mod group, CDHS group, and CC group underwent tumor-building experiments on day 11. Tumorigenicity was then assessed twice a week. After 4 weeks, feces, colon tissue, and tumor tissue were taken from the mice and were tested, and the mice were euthanized afterwards. Mice in the CDHS and CC groups showed symptoms similar to the clinical damp-heat syndrome observed in traditional Chinese medicine (TCM), and exhibited a worse general condition and more rapid tumor growth trend than those in the Mod group. The pathological examination indicated that inflammation was prevalent in the CDHS and CC groups. Both groups had a disrupted intestinal barrier and an overgrowth of pathogenic bacteria such as c_Gammaproteobacteria, o_Enterobacteriales, and g_Bacteroides. Their microbiota composition showed greater diversity. Intestinal flora may have a promising future in the discovery of indicators for evaluating a model of damp-heat syndrome in pancreatic cancer.

Rice Protein Peptides Alleviate Alcoholic Liver Disease via the PPARγ Signaling Pathway: Through Liver Metabolomics and Gut Microbiota Analysis.

Alcoholic liver disease (ALD) is the predominant type of liver disease worldwide, resulting in significant mortality and a high disease burden. ALD damages multiple organs, including the liver, gut, and brain, causing inflammation, oxidative stress, and fat deposition. In this study, we investigated the effects of rice protein peptides (RPP) on ALD in mice with a primary focus on the gut microbiota and liver metabolites. The results showed that administration of RPP significantly alleviated the symptoms of ALD in mice including adiposity, oxidative stress, and inflammation. The KEGG pathway shows that RPP downregulates the liver metabolite of capric acid and the metabolism of fatty acid biosynthesis compared with the MOD group. Mechanistically, RPP downregulated the PPARγ signaling pathway and suppressed the expression of fatty acid biosynthesis genes (FASN, ACC1, ACSL1, and ACSL3). Furthermore, two active peptides (YLPTKQ and PKLPR) with potential therapeutic functions for ALD were screened by Caco-2 cell modeling and molecular docking techniques. In addition, RPP treatment alleviates gut microbiota dysbiosis by reversing the F/B ratio, increasing the relative abundance of Alloprevotella and Alistipes, and upregulating the level of short-chain fatty acids. In conclusion, RPP alleviates ALD steatosis through the PPARγ signaling pathway by YLPTKQ and PKLPR and regulates gut microbiota.

"Evaluation of Silver Diamine Fluoride Modified Atraumatic Restorative Treatment (SMART) on hypomineralized first permanent molar"- a randomized controlled clinical study.

Restoring first permanent molars affected with molar incisor hypomineralization (MIH) is challenging. Focusing on improving the quality of life for children affected by MIH, at least until the complete eruption of first permanent molars to receive full coverage, to decrease the hypersensitivity and to be able to perform proper oral hygiene measures, the purpose of this study was to compare silver modified atraumatic restorative technique (SMART) versus the conventional restoration and fluoride varnish application on moderate to severe hypomineralized molars. The comparison considered the restoration survival, hypersensitivity, and digital surface area changes after one year follow up. Twenty-eight children were selected (20 girls and 8 boys) with at least 2 MIH molars with the same defect severity. The study comprised 2 groups; MOD group (moderate hypomineralized molar severity) and SEV group (Severe hypomineralized molar severity) (n = 28 tooth). Each group was further subdivided into 2 subgroups according to the technique of restoration: SMART subgroup and CONV subgroup (high viscosity glass ionomer restoration and fluoride varnish application) (each = 14 tooth). Evaluation was done in terms of the restoration survival (6 months and 12 months), hypersensitivity at 1 weak, 6 months and 12 months and occlusal surface area changes at 12 months). Professional Fluoride varnish application and home prophylaxis using MI paste were the protocol for each child patient. There was no significant difference between the 4 subgroups, regarding tooth restoration integrity at 6-months vs. 12-months. However, a statistically significant difference in tooth restoration integrity between the 4 subgroups at 12-months (P = .049). Also, the hypersensitivity score, there was a statistically significant difference between the 4-time intervals (P < .001) and a statistically significant difference in surface area changes between the 4 subgroups. Selective removal of carious tissue and SMART restoration, combined with dental home and professional preventive measures every 3 months maintained the integrity of restorations in severely and moderately affected permanent molars up to 1 year. The study protocol was retrospectively registered on Clinical Trials (NCT05931822-05/ 07/2023).

Effect of Solid-State Fermentation Products of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans on Growth Performance of Broilers and Prevention of Avian Colibacillosis.

This study investigates the impact of the solid-state fermentation products of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans (LCBs) on the growth characteristics, immune function, intestinal morphology, cecum microbial community, and prevention of avian colibacillosis in broilers. One hundred and twenty Hyland Brown broilers (aged one day) were divided randomly into three groups (four replicates of ten broilers per group). (1) The CON group was fed a basal diet. (2) The MOD group was fed a basal diet. On day 40, APEC strain SX02 (1.1 × 105 CFU/g) was administered to the breasts of chickens in this group. (3) The LCBs group was fed a basal diet supplemented with fermentation products (98.5% basal diet + 0.5% Lactobacillus plantarum and Candida utilis solid-state fermentation products + 1.0% Bacillus coagulans solid-state fermentation products). On day 40, the LCBs group received the same treatment as the MOD group. The experiment lasted 43 days. This study found that the average daily gain (ADG) of the LCBs group was significantly higher than that of the MOD group (p < 0.05), indicating that LCBs can significantly increase the ADG of broilers and improve the feed conversion ratio. Furthermore, compared to the MOD group, the heart bacterial load was significantly reduced in the LCBs group (p < 0.05), and the lesions less severe in the heart, liver, and jejunum were observed (p < 0.05). Additionally, the detection of intestinal flora showed a significant increase in the abundance of beneficial bacteria in the cecum of the LCBs group, while the number of Escherichia coli and Shigella decreased significantly. In conclusion, the solid fermentation of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans can improve the growth performance of broilers while also protecting against avian pathogenic Escherichia coli infection. This demonstrates the potential usefulness of these LCBs in feed production.

Neuroprotection with hypothermic reperfusion and extracorporeal cardiopulmonary resuscitation - A randomized controlled animal trial of prolonged ventricular fibrillation cardiac arrest in rats.

Extracorporeal cardiopulmonary resuscitation (ECPR) facilitates resuscitation with immediate and precise temperature control. This study aimed to determine the optimal reperfusion temperature to minimize neurological damage after ventricular fibrillation cardiac arrest (VFCA). Twenty-four rats were randomized (n = 8 per group) to normothermia (NT = 37°C), mild hypothermia (MH = 33°C) or moderate hypothermia (MOD = 27°C). The rats were subjected to 10 minutes of VFCA, before 15 minutes of ECPR at their respective target temperature. After ECPR weaning, rats in the MOD group were rapidly rewarmed to 33°C, and temperature maintained at 33°C (MH/MOD) or 37°C (NT) for 12 hours before slow rewarming to normothermia (MH/MOD). The primary outcome was 30-day survival with overall performance category (OPC) 1 or 2 (1 = normal, 2 = slight disability, 3 = severe disability, 4 = comatose, 5 = dead). Secondary outcomes included awakening rate (OPC ≤ 3) and neurological deficit score (NDS, from 0 = normal to 100 = brain dead). The survival rate did not differ between reperfusion temperatures (NT = 25%, MH = 63%, MOD = 38%, p = 0.301). MH had the lowest NDS (NT = 4[IQR 3-4], MH = 2[1-2], MOD = 5[3-5], p = 0.044) and highest awakening rate (NT = 25%, MH = 88%, MOD = 75%, p = 0.024). In conclusion, ECPR with 33°C reperfusion did not statistically significantly improve survival after VFCA when compared with 37°C or 27°C reperfusion but was neuroprotective as measured by awakening rate and neurological function.

The hemostatic activity and Mechanistic roles of glucosyloxybenzyl 2-isobutylmalate extract (BSCE) from Bletilla striata (Thunb.) Rchb.f. in Inhibiting pulmonary hemorrhage

BackgroundHemorrhagic events cause numerous deaths annually worldwide, highlighting the urgent need for effective hemostatic drugs. The glucosyloxybenzyl 2-isobutylmalates Control Extract (BSCE) from the orchid plant Bletilla striata (Thunb.) Rchb.f. has demonstrated significant hemostatic activity in both in vitro and in vivo studies. However, the effect and mechanism of BSCE on non-traumatic bleeding remain unclear. MethodsPulmonary hemorrhage was induced in 40 Sprague-Dawley rats by administering Zingiber officinale Roscoe. for 14 days. These rats were then randomly divided into five groups: model (Mod), positive control (YNBY), and BSCE low, medium, and high-dose groups. An additional 8 rats served as the control group (Con). The BSCE groups received different doses of BSCE for 10 days, while the YNBY group received Yunnan Baiyao suspension. The effects on body weight, food and water intake, red blood cell count (RBC), hemoglobin concentration (HGB), lung tissue pathology, platelet count, coagulation parameters, and fibrinolytic system markers were evaluated. Network pharmacology and molecular docking analyses were also conducted to identify potential targets and pathways involved in BSCE's effects. ResultsBSCE treatment significantly improved body weight, food intake, and water consumption in rats with pulmonary hemorrhage. RBC and HGB levels increased significantly in the BSCE medium and high-dose groups compared to the Mod group (P < 0.05). Pathological examination revealed that BSCE reduced lung tissue hemorrhage and inflammation, with improvements in alveolar structure. BSCE also positively affected platelet count, thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (FIB) levels, and fibrinolytic markers (D-dimer, PAI-1, and t-PA). Network pharmacology and molecular docking identified key targets such as MMPs, CASPs, and pathways including IL-17 and TNF signaling, suggesting BSCE's involvement in hemostasis and anti-inflammatory processes. ConclusionsBSCE exhibits significant hemostatic and protective effects on Z.officinale-induced pulmonary hemorrhage in rats by improving hematological parameters, reducing lung tissue damage, and modulating the coagulation and fibrinolytic systems. The study provides evidence supporting the potential of BSCE as a therapeutic agent for hemorrhagic diseases, with its efficacy linked to multi-target and multi-pathway interactions.

Melatonin alleviates brain injury in copper-laden rats: Underlying benefits for Wilson’s disease

Copper serves as an indispensable cofactor for all living organisms, and its excessive accumulation has been associated with a variety of diseases. Wilson’s disease (WD) serves as an illustrative example of copper toxicity in humans, frequently presenting with liver and/or neuropsychiatric symptoms. The current therapeutic drugs, penicillamine (PA) and zinc gluconate (ZnG), have constraints, and research on their combination efficacy remains insufficient. It has been reported that melatonin (MLT) plays a vital role in binding to transition metals and exhibits strong antioxidant capacity. To investigate the therapeutic efficacy of MLT and combined treatment, rats were randomly divided into the following seven groups: the control (Con) group, copper-laden model rat (Mod) group, PA-treated group, ZnG-treated group, MLT- treated group, PA-ZnG-treated group, and PA-MLT-treated group. Then potential mechanisms and targets were investigated using a combination of metabolomics and network pharmacology and verified by molecular docking and qPCR. The findings revealed that MLT and the combination significantly improved behavior, pathology and copper levels in copper-laden rats. The results of the metabolomics study showed that profoundly altered metabolites were identified, and alanine, aspartate and glutamate metabolism, pyruvate metabolism, citrate cycle (TCA cycle), and glycolysis/gluconeogenesis were explored. In addition, molecular docking showed that MLT had high binding affinity with key targets, and qPCR results revealed that MLT could reverse the mRNA expression of targets GOT2 and PKM2. It was concluded that MLT effectively improves brain injury in copper-laden rats, and this effect was linked with the altered features of the metabolite profiles.

The antidepressant effect of Shexiang Baoxin Pills on myocardial infarction rats with depression may be achieved through the inhibition of the NLRP3 inflammasome pathway.

Patients with myocardial infarction (MI) frequently experience a heightened incidence of depression, thereby increasing the risk of adverse cardiovascular events. Consequently, early detection and intervention in depressive symptoms among patients with MI are imperative. Shexiang Baoxin Pills (SBP), a Chinese patent medicine employed for the treatment of MI, exhibits diverse mechanisms targeting this condition. Nevertheless, its therapeutic efficacy on postmyocardial infarction depressive symptoms remains unclear. The aim of this study is to investigate the effectiveness and mechanism of SBP in managing depression during acute myocardial infarction (AMI). A rat model combining MI and depression was established, and the rats were randomly divided into four groups: the model (MOD) group, SBP group, Fluoxetine (FLX) group, and Sham group. After 28 days of drug intervention, cardiac function was assessed using echocardiography while behavior was evaluated through sucrose preference test (SPT), forced swimming test (FST), and open-field test (OFT). Additionally, levels of inflammatory factors in serum and hippocampus were measured along with NLRP3 inflammasome-related protein expression via Western blotting and immunofluorescence. SBP can enhance cardiac function in rats with AMI and depression, while significantly ameliorating depressive-like behavior. Compared to the Sham group, levels of IL-1β, IL-18, TNF-α, and other inflammatory factors were markedly elevated in the MOD group. However, expressions of these inflammatory factors were reduced to varying degrees following treatment with SBP or FLX. Analysis of NLRP3 inflammasome-related proteins in the hippocampus revealed a significant upregulation of IL-1β, IL-18, NLRP3, ASC, caspase-1, and GSDMD in the MOD group; conversely, these measures were significantly attenuated after SBP intervention. We have observed a significant amelioration in depression-like behavior upon SBP administration during the treatment of AMI, suggesting that this effect may be attributed to the inhibition of NLRP3-mediated pyroptosis. (The main findings are summarized in the graphical abstract in the supplementary file.).

Mechanism of tea polyphenols improving the sarcopenia in the aged type 2 diabetes model rats via mitochondrial quality control

To explore whether tea polyphenols(TP) improve sarcopenia in the aged type 2 diabetes(T2DM)model rats via mitochondrial quality control(MQC). A total of 55 2-month-old male SD rats were randomly divided into the control group(n=10), the aged model group(aged, n=10) and the aging T2DM model group(n=35). The aging T2DM model group rats were fed with high-sugar and high-fat diet and intraperitoneally injected with 50 mg/kg D-galactose daily. After 4 weeks, the aging T2DM model group rats were given a single intraperitoneal injection of 30 mg/kg streptozotocin(STZ). After STZ injection for 2 weeks, fasting blood glucose(FBG) ≥ 16.7 mmol/L was defined as successful T2DM model. When the model was successfully induced, the 30 model rats were randomly divided into aged T2DM group(Mod), 300 mg/kg TP teatment group(TP) and 3 mg/kg rosiglitazone treatment group(RSG) according to FBG, with 10 rats in each group. Each group was treated with 50 mg/kg D-galactose to induce senescence and fed with high glucose and fat for 8 weeks. Western blot was used to detect the expression of P53 protein in gastnemius muscle tissue of the model group at the end of the experiment, which was higher than that of the control group, indicating that the aging T2DM model was successfully established. FBG was detected by the blood glucose meter, gastnemius muscle relative weights was calculated, the microstructure of mitochondria of gastnemius muscle was observed by transmission electron microscope(TEM), the expression of mitochondrial biosynthesis-related proteins PGC-1α, mitochondrial dynamics-related proteins(OPA1, DRP1) and mitochondrial autophagy-related proteins(P62, LC3) in gastnemius muscle were detected by western blot. Compared with the control group, the level of FBG and the expression of P53 in the Mod group were increased(P&lt;0.01). The gastnemius muscle relative weights, the expression level of PGC-1α, OPA1 and the ratio of LC3II/LC3I were decreased(P&lt;0.01). The expression level of P62 and DRP1 were significantly increased(P&lt;0.01). The number of mitochondria decreased, the volume decreased and a large number of vacuolization, and there were no obvious autophagolysosomes and fission and fusion. After 8 weeks, compared with the Mod group, the number of mitochondria in the gastrocnemius of TP and RSG groups, vacuolization, fission and fusion were improved, and the autophagolysosomes was significantly increased. The expression levels of P53, DRP1 and P62, the level of FBG in the TP group were significantly decreased(P&lt;0.01, P&lt;0.05). The expression levels of OPA1 and PGC-1α, the ratios of LC3II/LC3I and gastnemius muscle relative weights were significantly increased(P&lt;0.05, P&lt;0.01). TP can improve the sarcopenia in the aged T2DM model rats, and its mechanism is related to the regulation of mitochondrial quality control.

Management of Cowlick in Female Hairline Correction Surgery.

To investigate the results of two grafting methods to manage CLs in the FHL of women. The present study was performed to investigate the results of two grafting methods to manage cowlicks (CLs) in the frontal hairline of women. Patients were categorized into two groups based on the grafting method: the matching with the original hair direction (MOD) group and the matching with the CL hair direction (MCD) group. Satisfaction ratings were compared between the two groups. Among 80 patients, 77 had a single CL and 3 had a double CL. In the MOD group, 7 of 12 (58.3%) patients underwent a touch-up procedure for density enhancement with an average of 123 follicular units; the average patient and physician satisfaction ratings were 4.84 and 4.60, respectively. In the MCD group, 7 of 68 (10.3%) patients underwent a touch-up procedure for density enhancement around the CL; the average patient and physician satisfaction ratings were 4.64 and 4.50, respectively. In female hairline correction surgery, the surgeon must choose between MOD and MCD by considering the location, shape, and pattern of the CL; the facial shape; and the patient's preference. Delicate graft placement is required to obtain natural-looking results. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors. www.springer.com/00266 .

Whole Transcriptome Sequencing of Peripheral Blood Identifies the Alzheimer's Disease-Related circRNA-miRNA-lncRNA Pathway.

Previous studies on transcriptional profiles suggested dysregulation of multiple RNA species in Alzheimer's disease. However, despite recent investigations revealing various aspects of circular RNA (circRNA)-associated competing endogenous RNA (ceRNA) networks in Alzheimer's Disease (AD) pathogenesis, few genome-wide studies have explored circRNA-associated profiles in AD patients exhibiting varying degrees of cognitive loss. To investigate the potential pathogenesis-related molecular biological changes in the various stages of AD progression. Whole transcriptome sequencing was performed on the peripheral blood of 7 normal cognition (NC) subjects, 8 patients with mild cognitive impairment, 8 AD patients with mild dementia (miD), and 7 AD patients with moderate dementia (moD). Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were conducted to predict the potential functions of the maternal genes of microRNAs (miRNAs), circRNAs and long non-coding RNAs (lncRNAs). The construction of ceRNA network was performed between the NC group and each diseased group based on the differently expressed RNAs. In total, 3568 messenger RNAs (mRNAs), 142 miRNAs, 990 lncRNAs, and 183 circRNAs were identified as significantly differentially expressed across the four groups. GO and KEGG enrichment analysis revealed the significant roles of GTPase activity and the MAPK signaling pathway in AD pathogenesis. A circRNA-miRNA-lncRNA ceRNA pathway, characterized by the downregulated hsa-miR-7-5p and upregulated hsa_circ_0001170, was identified based on the differentially expressed RNAs between the NC group and the moD group. The study suggests that circRNAs may be independent of mRNAs in AD pathogenesis and holds promise as potential biomarkers for AD clinical manifestations and pathological changes.

Short-duration aerobic high-intensity intervals versus moderate exercise training intensity in patients with peripheral artery disease: study protocol for a randomised controlled trial (the Angiof-HIIT Study)

IntroductionSupervised exercise training is among the first-line therapies for patients with peripheral artery disease (PAD). Current recommendations for exercise include guidance focusing on claudication pain, programme and session duration, and...

Study on miRNA expression and regulatory network of Amygdalus mongolica oil and amygdalin in the treatment of pulmonary fibrosis

IntroductionAmygdalus mongolica (Maxim.) Ricker is a traditional Chinese medicine known for its ability to relieve cough and expectorate phlegm and is commonly used to treat dry throat, dry cough, and bronchitis. A. mongolica oil and amygdalin were the active components isolated from the seed kernel seeds of A. mongolica. The main objective of this study was to investigate the effects of A. mongolica oil and amygdalin on bleomycin-induced pulmonary fibrosis (PF) in rats by integrating pharmacodynamic, transcriptomics, and bioinformatics, to determine the changes of miRNAs in both against PF, and to explore the underlying mechanisms. MethodsA rat model of PF was induced by administering bleomycin through the trachea. The rats were randomized into several groups: a control group (CON), an untreated model group (MOD), an active control group treated with pirfenidone (PFD), and groups receiving different doses of A. mongolica oil (5 g/kg [OIL-L], 10 g/kg [OIL-M], and 15 g/kg [OIL-H]), or amygdalin (20 mg/kg [AMY-L], 40 mg/kg [AMY-M], and 80 mg/kg [AMY-H]). After 28 days, samples were collected for subsequent pharmacodynamic and transcriptomic analyses. ResultsThe results demonstrated that A. mongolica oil and amygdalin effectively reduced the abnormally elevated levels of IL-1β, IL-6, HA, LN and COL-IV in PF rats and alleviated bleomycin-induced pulmonary fibrotic changes. Furthermore, compared to the MOD group, the expression levels of 27 miRNAs were up-regulated and 23 miRNAs were down-regulated in the OIL 10 g/kg group. Among the up-regulated miRNAs, two novel differentially expressed miRNAs (DE-miRNAs) were identified. In the AMY 20 mg/kg group, 12 miRNAs were up-regulated and 10 were down-regulated when compared to the MOD group. These DE-miRNAs were found to be enriched in pathways such as the PI3K-Akt signalling pathway, MAPK signalling pathway, and calcium signalling pathway. The microRNAs miR-301b-3p, miR-667-5p, miR-363-3p, and miR-540-3p may be associated with both the anti-PF mechanism of A. mongolica oil and the anti-PF mechanism of amygdalin. DiscussionNumerous studies have demonstrated the significance of various pathways, including PI3K-Akt, MAPK, and calcium signalling pathways, in the mechanism of PF and the development of anti-PF medications. Meanwhile, multiple miRNAs, including miR-301b-3p, miR-667-5p, miR-363-3p, and miR-540-3p, have been found to be associated with lung disease and fibrosis. ConclusionsThe present experiments confirmed the effectiveness of A. mongolica oil and amygdalin in reducing inflammation and fibrosis in bleomycin-induced PF rats, with particularly favorable outcomes observed in the A. mongolica oil 10 g/kg versus amygdalin 20 mg/kg. Meanwhile, miRNAs miR-301b-3p, miR-667-5p, miR-363-3p and miR-540-3p were found to be potentially associated with both A. mongolica oil and amygdalin anti-PF mechanisms. Based on our findings, we suggest that these miRNAs might play key roles in the anti-PF mechanism of A. mongolica. The individual miRNAs require further investigation.

Huayu Qutan Recipe promotes lipophagy and cholesterol efflux through the mTORC1/TFEB/ABCA1-SCARB1 signal axis.

This study aims to investigate the mechanism of the anti-atherosclerosis effect of Huayu Qutan Recipe (HYQT) on the inhibition of foam cell formation. Invivo, the mice were randomly divided into three groups: CTRL group, MOD group and HYQT group. The HYQT group received HYQT oral administration twice a day (20.54 g/kg/d), and the plaque formation in ApoE-/- mice was observed using haematoxylin-eosin (HE) staining and oil red O (ORO) staining. The co-localization of aortic macrophages and lipid droplets (LDs) was examined using fluorescent labelling of CD11b and BODIPY fluorescence probe. Invitro, RAW 264.7 cells were exposed to 50 μg/mL ox-LDL for 48 h and then treated with HYQT for 24 h. The accumulation of LDs was evaluated using ORO and BODIPY. Cell viability was assessed using the CCK-8 assay. The co-localization of LC3b and BODIPY was detected via immunofluorescence and fluorescence probe. LysoTracker Red and BODIPY 493/503 were used as markers for lysosomes and LDs, respectively. Autophagosome formation were observed via transmission electron microscopy. The levels of LC3A/B II/LC3A/B I, p-mTOR/mTOR, p-4EBP1/4EBP1, p-P70S6K/P70S6K and TFEB protein level were examined via western blotting, while SQSTM1/p62, Beclin1, ABCA1, ABCG1 and SCARB1 were examined via qRT-PCR and western blotting. The nuclear translocation of TFEB was detected using immunofluorescence. The components of HYQT medicated serum were determined using Q-Orbitrap high-resolution MS analysis. Molecular docking was employed to identify the components of HYQT medicated serum responsible for the mTOR signalling pathway. The mechanism of taurine was illustrated. HYQT has a remarkable effect on atherosclerotic plaque formation and blood lipid level in ApoE-/- mice. HYQT decreased the co-localization of CD11b and BODIPY. HYQT (10% medicated serum) reduced the LDs accumulation in RAW 264.7 cells. HYQT and RAPA (rapamycin, a mTOR inhibitor) could promote cholesterol efflux, while chloroquine (CQ, an autophagy inhibitor) weakened the effect of HYQT. Moreover, MHY1485 (a mTOR agonist) also mitigated the effects of HYQT by reduced cholesterol efflux. qRT-PCR and WB results suggested that HYQT improved the expression of the proteins ABCA1, ABCG1 and SCARB1.HYQT regulates ABCA1 and SCARB1 protein depending on the mTORC1/TFEB signalling pathway. However, the activation of ABCG1 does not depend on this pathway. Q-Orbitrap high-resolution MS analysis results demonstrated that seven core compounds have good binding ability to the mTOR protein. Taurine may play an important role in the mechanism regulation. HYQT may reduce cardiovascular risk by promoting cholesterol efflux and degrading macrophage-derived foam cell formation. It has been observed that HYQT and ox-LDL regulate lipophagy through the mTOR/TFEB signalling pathway, rather than the mTOR/4EBP1/P70S6K pathway. Additionally, HYQT is found to regulate cholesterol efflux through the mTORC1/TFEB/ABCA1-SCARB1 signal axis, while taurine plays a significant role in lipophagy.

Integrated UHPLC-MS untargeted metabolomics and gut microbe metabolism pathway-targeted metabolomics to reveal the prevention mechanism of Gushudan on kidney-yang-deficiency-syndrome rats

Gushudan (GSD) was a traditional Chinese prescription with the remarkable effect of kidney-tonifying and bone-strengthening. However, the potential prevention mechanisms of the GSD on kidney-yang-deficiency-syndrome (KYDS) and its regulation on gut microbe metabolism still need to be further systematically investigated. This study established untargeted urinary metabolomics based on RP/HILIC-UHPLC-Q-Orbitrap HRMS and combined with multivariate statistical analysis to discover differential metabolites and key metabolic pathways. And the gut microbe metabolism pathway-targeted metabolomic based on HILIC-UHPLC-MS/MS was developed and validated to simultaneously determine 15 gut microbe-mediated metabolites in urine samples from the control group (CON), KYDS model group (MOD), GSD-treatment group (GSD) and positive group (POS). The results showed that a total of 36 differential metabolites were discovered in untargeted metabolomics. These differential metabolites included proline, cytosine, butyric acid and nicotinic acid, which were primarily involved in the gut microbe metabolism, amino acid metabolism, energy metabolism and nucleotide metabolism. And GSD played a role in preventing KYDS by regulating these metabolic pathways. The targeted metabolomics found that the levels of 10 gut microbe-mediated metabolites had significant differences in different groups. Among them, compared with the CON group, the levels of lysine, tryptophan, phenylacetylglycine and hippuric acid were increased in the MOD group, while the levels of threonine, leucine, dimethylamine, trimethylamine, succinic acid and butyric acid were decreased, which verified the disorders of gut microbe metabolism in the KYDS rats and GSD had a significant regulatory effect on this disorder. As well as by comparing analysis, it was found that the experimental results were consistent with previous metabolomics and microbiomics of fecal samples. Therefore, this integrated strategy of untargeted and targeted metabolomics not only elucidated the potential prevention mechanism of GSD on KYDS, but also provided a scientific basis for GSD preventing KYDS via the “gut-kidney” axis.

Anwulignan Alleviates the Acute Liver Injury Induced by Acetaminophen in Mice

Background Drug-induced liver injury (DILI) is extremely common in clinical practice and needs to be focused. Anwulignan is a unique lignan monomer of Schisandra Sphenanthera Fructus and is used as the main active ingredient of Wuzhi tablet, which is used for liver protection in clinical practice. In this study, an acute liver injury mouse model induced by acetaminophen and paracetamol (APAP) was used to observe its effects. Materials and Methods An acute liver injury mouse model was established by intraperitoneally injecting APAP. Mice were divided into blank control (CON) group, APAP model (MOD) group, Anwulignan CON group, and Anwulignan MOD group. Anwulignan (6 mg/kg) or sodium carboxymethylcellulose (10 mL/kg) was intragastrically given to mice once a day, successively for 14 days. On day 14, 250 mg/kg APAP solution (10 mL/kg) or saline was injected intraperitoneally, and 12 h later, the mice were killed. alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the serum of mice were tested. The liver index of mice was calculated, and the pathology of liver injury was evaluated by HE staining in mice. The activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) as well as the content of malondialdehyde (MDA) in the liver tissue were tested. The levels of kelch-like ECH-associated protein 1 (Keap-1), nuclear factor erythroid-2-related factor 2 (Nrf2), heme oxygenase 1 (HO-1), BCL2 associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), caspase-3 and tumor suppressor protein (P53) mRNAs in the liver tissues of mice were detected by reverse transcription-PCR (RT-PCR), and the expression levels of Keap-1, Nrf2, HO-1, Bax, Bcl-2, caspase-3 and P53 proteins in the liver tissue of mice were detected by Western blot. Results Anwulignan can alleviate the acute liver injury of mice. The down-regulation of the cyclophosphamide (CYP2E1) expression in the liver tissue of mice to reduce the hepatotoxicity of APAP, and the regulation of Nrf2-ARE signaling pathway-related gene expressions to play an antioxidant role and apoptosis-related gene expressions to inhibit the liver cell apoptosis may be involved in the mechanism through which Anwulignan can exert its effect. Conclusion Anwulignan can alleviate the liver injury induced by APAP in mice, and this study may provide evidence for researching and developing some drugs and nutraceuticals used for preventing and treating liver injury.

Alterations of the gut microbiota and metabolites by ShenZhu TiaoPi granule alleviates hyperglycemia in GK rats.

ShenZhu TiaoPi granule (STG) is a compound prescription that is used in Chinese medicine for the treatment of type 2 diabetes mellitus (T2DM). Previous studies have indicated a hypoglycaemic effect, but the underlying mechanism remains unclear. Goto-Kakizaki (GK) rats were used to establish an in vivo T2DM model (Mod). The metformin (Met) and STG treatment time was 12 weeks. Fasting blood glucose (FBG) and insulin levels and the area under the glucose curve (GAUC) were measured. Intestinal pathology and permeability were observed. Microbial diversity analysis and metabolomics were used to investigate the underlying mechanisms. Compared with the Con group, the T2DM Mod group presented significant differences in weight, FBG, GAUC, and homeostasis model assessment-insulin resistance (HOMA-IR) indices (p < 0.01). Met and STG improved these indicators (p < 0.01). The pathological morphology and zonula occludens 1 protein levels in the intestines of the Mod group of rats were altered, leading to increases in the lipopolysaccharide (LPS) and interleukin-1β (IL-1β) levels. In the Met and STG groups, the intestinal conditions improved, and the LPS and IL-1β levels significantly decreased (p < 0.01). Changes in the gut microbiota and metabolites occurred in the Mod group. In the STG group, the abundance of Intestinimonas increased, and the abundance of Eubacterium coprostanoligenes decreased significantly (p < 0.05). Moreover, STG also altered 2-deoxyglucose, beta-muricholic acid and dioxolithocholic acid production. In addition, the main metabolic pathways affected by STG were bile acid biosynthesis and cholesterol metabolism. Intestinimonas, D-maltose_and_alpha-lactose may be potential biomarkers for the effects of STG. STG alleviates hyperglycaemia via the gut microbiota and metabolites in GK rats.

Effects and mechanisms of Salvia miltiorrhiza Bunge extract on myocardial cell apoptosis in rat heart failure model.

This work aimed to investigate the effects of Tanshinone IIA (Tan IIA) on myocardial cell (MC) apoptosis in a rat model of heart failure (HF). Tan IIA was extracted from Salvia miltiorrhiza Bunge (SMB) using an ethanol reflux method. Fifty rats were randomly divided into five groups: sham (no treatment), mod (HF model establishment), low dose (LD: 0.1 mL/kg Tan IIA), medium dose (MD: 0.3 mL/kg Tan IIA), and high dose (HD: 0.5 mL/kg Tan IIA), with 10 rats in each group. The effects of different doses of Tan IIA on cardiac function, MC apoptosis, and the levels of proteins associated with the PI3K/Akt/mTOR signaling pathway were compared. Mod group showed a significant decrease in systolic arterial pressure, mean arterial pressure, heart rate, left ventricular systolic pressure, left ventricular ejection fraction, left ventricular fractional shortening, and the levels of p-PI3K, p-Akt, and p-mTOR proteins versus sham group (p < 0.05). Additionally, the left ventricular end-diastolic diameter (LVIDd), end-systolic diameter, diastolic pressure, and MC apoptosis were significantly increased (p < 0.05). LD, MD, and HD groups exhibited significant improvements across various indicators of cardiac function and MC apoptosis versus mod group (p < 0.05). Tan IIA may improve cardiac function and inhibit MC apoptosis in rats with HF by modulating the PI3K/Akt/mTOR signaling pathway.

Impact of Access Cavity Design on Fracture Resistance of Endodontically Treated Maxillary First Premolar: In Vitro.

This study was designed to investigate the impact of access cavity designs on fracture resistance of endodontically treated maxillary first premolars. The study sample consisted of 72 intact maxillary first premolars, randomly divided into six groups (n = 12). A standardized proximal cavity preparation was prepared for all samples using standard bur. Groups I: control group with only standard proximal cavity and no endodontic access, group II: Truss access cavity, group III: Separated access to buccal and palatal canals without removal of dentine in between, group IV: Access to buccal and palatal canals with removal of dentine in between, group V: Traditional access cavity, group VI: Mesio-occlusal-distal cavity (MOD). For groups I and VI, only composite restoration was used to restore the proximal cavity, while for groups II- V, the access was prepared and endodontic treatment was performed on all teeth, then composite restoration was placed. The root canals were instrumented using nickel-titanium files, irrigated with sodium hypochlorite, and filled with AH plus sealer and gutta-percha using warm vertical condensation. All samples were then placed in an acrylic mold and underwent thermal aging for 10,000 cycles between 5 and 55°C. The samples were fixed in a universal testing machine with the long axis of the roots positioned at 20° to a load applied at a crosshead speed of 1 mm/min using a stainless steel semi-spherical indenter (Ø = 3 mm) until fracture occurred to determine the fracture resistance force in Newton. The normality test (Shapiro-Wilk) showed that data are normally distributed. Group II exhibited the highest mean fracture resistance, and group VI was the least likely to resist the fracture. No statistically significant differences between tested groups (p-value = 0.237). The MOD group showed a more unfavorable mode of fracture compared to other groups. No significant difference in fracture resistance between conservative and traditional access cavities. The missing marginal ridges, such as in MOD cavities played an important role in decreasing the fracture resistance of endodontically treated teeth.

Exploring the effects and mechanism of peony pollen in treating benign prostatic hyperplasia

Paeonia suffruticosa is widely cultivated globally due to its medicinal and ornamental value. Peony pollen (PP) is commonly used in Chinese folk medicine to make tea to treat benign prostatic hyperplasia (BPH), but its molecular mechanism against BPH is yet to be comprehended. The objective of this research was to experimentally verify the effect of PP in the treatment of BPH and to preliminarily reveal its mechanism of action on BPH using network pharmacology methods. The results revealed that PP could decrease prostate volume and prostate index, serum testosterone (T), dihydrotestosterone (DHT), and estradiol (E2) levels. Moreover, it could improve prostate tissue structure in BPH model animals as well. Additionally, database searches and disease target matching revealed 81 compounds in PP. Of these, 3, 7, 8, 2′-tetrahydroxyflavone, Chrysin, Wogonin, Limocitrin, and Sexangularetin were the top five compounds associated with the therapeutic effects of BPH. Furthermore, 177 therapeutic targets for BPH were retrieved from databases of Swiss Target, DisGeNET, Drugbank, Genecards, OMIM, TTD, and Uniprot. In contrast, core targets AKT1, EGFR, IL6, TNF, and VEGFA were obtained by PPI network diagram. Molecular docking also showed that the main efficacy components and potential core targets in PP had good binding capacity. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomics (KEGG) analysis established that the effect of PP in BPH therapy was mainly through regulating the expression levels of protein kinase B on phosphatidylinositol 3-kinase and phosphatidylinositol 3-kinase-protein kinase B pathways. Additionally, Western blot experiments also exhibited a significant elevation in the activated PI3K and AKT proteins in the model (Mod) group relative to the control (Con) group, and the expression of these activated proteins was significantly reduced after PP administration. In summary, this research provides a scientific basis for employing PP to treat BPH, preliminarily reveals its mechanism of action and potential targets, and lays the foundation for further research and development.