Gut microbiota regulates the ALK5/NOX1 axis by altering glutamine metabolism to inhibit ferroptosis of intrahepatic cholangiocarcinoma cells.

Intrahepatic cholangiocarcinoma (ICC) is a kind of hepatobiliary tumor that is increasing in incidence and mortality. The gut microbiota plays a role in the onset and progression of cancer, however, the specific mechanism by which the gut microbiota acts on ICC remains unclear. In this study, feces and plasma from healthy controls and ICC patients were collected for 16S rRNA sequencing or metabolomics analysis. Gut microbiota analysis showed that gut microbiota abundance and biodiversity were altered in ICC patients compared with controls. Plasma metabolism analysis showed that the metabolite glutamine content of the ICC patient was significantly higher than that of the controls. KEGG pathway analysis showed that glutamine plays a vital role in ICC. In addition, the use of antibiotics in ICC animals further confirmed that changes in gut microbiota affect changes in glutamine. Further experiments showed that supplementation with glutamine inhibited ferroptosis and downregulated ALK5 and NOX1 expression in HuCCT1 cells. ALK5 overexpression or NOX1 overexpression increased NOX1, p53, PTGS2, ACSL4, LPCAT3, ROS, MDA and Fe2+ and decreased FTH1, SLC7A11 and GSH. Knockdown of NOX1 suppressed FIN56-induced ferroptosis. In vivo, supplementation with glutamine promoted tumor growth. Overexpression of ALK5 repressed tumor growth and induced ferroptosis in nude mice, which could be reversed by the addition of glutamine. Our results suggested that the gut microbiota altered glutamine metabolism to inhibit ferroptosis in ICC by regulating the ALK5/NOX1 axis.

The Efficacy of Traditional Chinese Herbal Medicine Across Multiple Cardiovascular Diseases: An Umbrella Review of Systematic Reviews of Randomized Controlled Trials.

ABSTRACT: Traditional Chinese herbal medicine (CHM) has been extensively used in cardiovascular disease (CVD) in modern clinical practice, alone or in combination with conventional treatment. However, its efficacy has not been assessed extensively. From inception until August 2023, we systematically searched 5 public literature databases to conduct the umbrella review. The inclusion criterion is systematic reviews of randomized controlled trials investigating the effect of CHM in the contemporary management of CVDs. The quality of the included systematic reviews, the certainty of the evidence, and the potential risk of bias were assessed. Five hundred and thirty-nine systematic reviews, including 346 studies in Chinese and 193 in English, were selected before the quantitative synthesis. The methodological quality was generally moderate, with a median value of 11. The favorable efficacy of CHM was primarily presented on 5 main conditions: coronary artery disease, hypertension, heart failure, restenosis, and angina pectoris. CHM, with or without conventional treatment, showed a consistent beneficial effect in various CVDs. Nevertheless, the magnitude of the effect requires further investigation as the lack of relevant research and the complexity of the clinical practice of CHM.

Visual analysis of acupuncture point selection patterns and related mechanisms in acupuncture for hypertension.

BACKGROUND: Hypertension has become one of the most pathogenic diseases in the world. OBJECTIVE: This paper summarizes and analyzes the acupuncture point combinations and treatment principles of acupuncture for hypertension in a systematic way by means of big data mining. METHODS: The literature for this paper was obtained from CNKI, Wanfang, VIP, SinoMed and PubMed, Embase, Cochrane Library, Web of Science, and Ovid databases. Thedata were collected to obtain combinations of acupoints with strong associations through association rule analysis, complex networks for screening to obtain core acupoint nuclei, and cluster analysis to derive treatment principles. RESULTS: A total of 127 acupuncture prescriptions involving 66 acupoints were included in this study. Tai-chong (LR3), Qu-chi (LI11), Zu-san-li (ST36), Feng-chi (GB20), and He-gu (LI4) were the most commonly used acupoints. The large intestine meridian was the preferred meridian, and most of the extremity acupoints, especially the lower extremities, were selected clinically. The association rule reveals that Qu-chi (LI11) and Zu-san-li (ST36) are the dominant combination acupoints. 3 core association points obtained after complex network analysis, the 1st association, Bai-hui (DU20), Tai-xi (KI3), Gan-shu (BL18), Shen-shu (BL23); The 2nd association, Qu-chi (LI11), He-gu (LI4), San-yin-jiao (SP6), Zu-san-li (ST36), Feng-chi (GB20), Tai-chong (LR3); The 3rd association, Qi-hai (RN6), Guan-yuan (RN4), Zhong-wan (RN12), Zhao-hai (KI6), Tai-yang (EX-HN5), Lie-que (LU7), Yang-ling-quan (GB34), Xing-jian (LR2), Yin-ling-quan (SP9). Cluster analysis yielded the treatment principles of nourishing Yin and submerging Yang, pacifying the liver and submerging Yang, tonifying Qi and Blood, and calming the mind and restoring the pulse, improving clinical outcomes. CONCLUSION: By means of big data mining, we can provide reference for acupuncture point grouping and selection for clinical acupuncture treatment of hypertension.

Effect of Grape Seed Procyanidins Combined with Allicin on Lipid Levels in Hyperlipidemic Rats.

OBJECTIVES: To study the effects of grape seed proanthocyanidins (GSP) combined with allicin on serum lipids level and vascular damage in a rat model of hyperlipidemia. MATERIALS AND METHODS: SD rats(male, 170-220 gn= 40) were randomized into five groups (n = 8/group): modelhigh fat and cholesterol diet; controlnormal diet; model+low-dose (GSP+allicin )(GSP 45mg/kg, allicin 30mg/kg, orally); model+high-dose (GSP+allicin) (GSP180mg/kg, allicin 90mg/kg, orally) and positive control (model+simvastatin (4 mg/kg)). Normal control group was fed conventionally, and remaining four groups were fed high cholesterol and fat food to replicate the high fat model. After 9 weeks, the normal control group continued to receive regular feeding, while the other groups continued to receive high-fat feeding. At the same time, model and normal control groups were given equal volume of physiological saline by gavage, and the other treatment groups began to receive corresponding drugs by gavage once a day. After 4 weeks, serum levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) as well as high-density lipoprotein cholesterol (HDL-C) in rats were determined. And the body weight of rat, total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and malondialdehyde (MDA)in serum were identified. The level of endothelin-1(ET-1) was quantitative analysis by ELISA assay. RESULTS: In comparison to normal controls, the model group displayed a marked rise in body weight, an increment in serum concentrations of LDL-C, TG and TC, as well as a decline in HDL (P<0.01), demonstrating successful model replication; All doses of GSP in combination with allicin resulted in a reduction in TG, LDL-C, and TC and an enhancement in HDL-C in contrast to the model control (all P<0.05). High-dose (GSP+allicin ) decreased MDA, and increased T-AOC and SOD activity(all P<0.01). All doses of GSP combined with allicin decreased ET-1 (all P<0.05). In addition, the protective effect of GSP combined with allicin was dose-dependent. CONCLUSIONS: Studies have shown that GSP combined with allicin can significantly improve blood lipids in hyperlipidemic rats, and this mechanism may be related to antioxidants and reduced endothelial damage.

Clinical and psychological status analysis of children and parents infected with familial aggregation omicron variant in Shanghai in parent-child ward.

Aims: To analyze the clinical characteristics, treatment outcomes and sleep psychological problems of children and parents infected with familial aggregation Omicron variants under a parent-child ward treatment mode to provide a theoretical reference for the diagnosis and comprehensive treatment of Omicron variant strains. Methods: The clinical data of 225 children and 230 adult family members admitted were retrospectively collected and analyzed to investigate their clinical characteristics and response to treatments. Results: The proportion of infected adults and children was the same, and the proportion of children with mild disease was higher than that of adults, but the clinical symptoms were milder. The clinical symptoms of fever, nausea, vomiting and wheezing in children were significantly higher than in adults (P < 0.05). In addition, dry pharynx, pharynx itching and pharyngeal pain were lower than in adults (P < 0.05). The time of turning negative in the moderate group was longer than in the mild group, and the time of turning negative in the unvaccinated group was higher than in the vaccinated group (P < 0.05). The Cycle Threshold Value (Ct value) of Open Reading Frame 1ab (ORF1ab) and Nucleocapsid protein (N) gene of children were higher adults. The increase in the rate of Ct value of ORF1ab and N gene in adults treated with Traditional Chinese Medicine (TCM) was significantly higher than in those who underwent symptomatic treatment (P < 0.01). Based on the Children's Sleep Habits Questionnaire (CSHQ)score, we found varying levels of sleep problems in sleeping habits, latency and anxiety, night awakenings and abnormal sleep at all ages (P < 0.05). In the adult group, those with Self-Rating Scale of Sleep (SRSS) scores ≥3 accounted for more than 50% of adults with insomnia, sleep deprivation, sleep instability and early awakening. The proportion of adults with anxiety and depression was 21.3% and 16.4%. Conclusion: Infections in children and adults during this pandemic were mainly associated with familial aggregation infections, and their clinical symptoms were mainly located in the upper respiratory tract. With comprehensive treatment, children became negative faster, vaccination led to faster recovery, and although some patients experienced sleeping and psychological issues, all patients had good prognoses following comprehensive diagnosis under a parent-child ward treatment mode.

[Clinical features of children and their family members with family clusters of SARS-CoV-2 Omicron variant infection in Shanghai, China: an analysis of 380 cases]. / 上海380例家庭聚集性感染Omicronå˜å¼‚æ ªçš„å„¿ç«¥åŠå ¶å®¶å±žä¸´åºŠç‰¹å¾åˆ†æž.

OBJECTIVES: To study the clinical features and prognosis of children and their family members with family clusters of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant infection under the admission mode of parent-child ward. METHODS: A retrospective analysis was performed on the medical data of 190 children and 190 family members with SARS-CoV-2 Omicron variant infection who were admitted to Shanghai Sixth People's Hospital, the designated hospital for coronavirus disease 2019 (COVID-19), April 8 to May 10, 2022. RESULTS: Both the child and adult groups were mainly mild COVID-19, and the proportion of mild cases in the child group was higher than that in the adult group (P<0.05). Respiratory symptoms were the main clinical manifestations in both groups. Compared with the adult group, the child group had higher incidence rates of fever, abdominal pain, diarrhea, and wheezing (P<0.05) and lower incidence rates of nasal obstruction, runny nose, cough, dry throat, throat itching, and throat pain (P<0.05). Compared with the child group, the adult group had higher rates of use of Chinese patent drugs, traditional Chinese medicine decoction, recombinant interferon spray, cough-relieving and phlegm-eliminating drugs, and nirmatrelvir/ritonavir tablets (P<0.05). Compared with the adult group, the child group had a lower vaccination rate of SARS-CoV-2 vaccine (30.5% vs 71.1%, P<0.001) and a shorter duration of positive SARS-CoV-2 nucleic acid (P<0.05). The patients with mild COVID-19 had a shorter duration of positive SARS-CoV-2 nucleic acid than those with common COVID-19 in both groups (P<0.05). The patients with underlying diseases had a longer duration of positive SARS-CoV-2 nucleic acid than those without such diseases in both groups (P<0.05). CONCLUSIONS: Both children and adults with family clusters of SARS-CoV-2 Omicron variant infection manifest mainly mild COVID-19. Despite lower vaccination rate of SARS-CoV-2 vaccine in children, they have rapid disease recovery, with a shorter duration of positive SARS-CoV-2 nucleic acid than adults, under the admission mode of parent-child ward.

Multifunctional Doxorubicin@Hollow-Cu9S8 nanoplatforms for Photothermally-Augmented Chemodynamic-Chemo therapy.

Multimodal therapy has attracted increasing interests in tumor treatment due to its high anti-cancer efficacy, and the key is to develop multifunctional nanoagents. The classic multifunctional nanoagents are made up of expensive and complex components, leading to limited practical applications. To solve these problems, we have developed the polyethylene glycol (PEG) coated hollow Cu9S8 nanoparticles (H-Cu9S8/PEG NPs), whose H-Cu9S8 component exhibits the photothermal effect for near-infrared (NIR) photothermal therapy (PTT), the Fenton-like catalytic activity for chemodynamic therapy (CDT), and the drug-loading capacity for chemotherapy. The H-Cu9S8/PEG NPs with a diameter of âˆ¼ 100 nm have been synthesized by sulfurizing cuprous oxide (Cu2O) nanoparticles through "Kirkendall effect", and they exhibit high photothermal conversion efficiency of 40.9%. Meanwhile, the H-Cu9S8/PEG NPs are capable of a Fenton-like reaction, which can be augmented by 2 times under the NIR irradiation. The hollow structure gives the H-Cu9S8/PEG high doxorubicin (DOX) loading capacity (21.1%), and then the DOX release can be further improved by pH and photothermal effect. When the DOX@H-Cu9S8/PEG dispersions are injected into the tumor-bearing mice, the tumor growth can be efficiently inhibited due to the synergistic effect of photothermally-augmented CDT-chemo therapy. Therefore, the DOX@H-Cu9S8/PEG can serve as a multifunctional nanoplatform for photothermally-augmented CDT-chemo treatment of malignant tumors.

Mechanisms of motor symptom improvement by long-term Tai Chi training in Parkinson's disease patients.

BACKGROUND: Tai Chi has been shown to improve motor symptoms in Parkinson's disease (PD), but its long-term effects and the related mechanisms remain to be elucidated. In this study, we investigated the effects of long-term Tai Chi training on motor symptoms in PD and the underlying mechanisms. METHODS: Ninety-five early-stage PD patients were enrolled and randomly divided into Tai Chi (n = 32), brisk walking (n = 31) and no-exercise (n = 32) groups. At baseline, 6 months and 12 months during one-year intervention, all participants underwent motor symptom evaluation by Berg balance scale (BBS), Unified PD rating-scale (UPDRS), Timed Up and Go test (TUG) and 3D gait analysis, functional magnetic resonance imaging (fMRI), plasma cytokine and metabolomics analysis, and blood Huntingtin interaction protein 2 (HIP2) mRNA level analysis. Longitudinal self-changes were calculated using repeated measures ANOVA. GEE (generalized estimating equations) was used to assess factors associated with the longitudinal data of rating scales. Switch rates were used for fMRI analysis. False discovery rate correction was used for multiple correction. RESULTS: Participants in the Tai Chi group had better performance in BBS, UPDRS, TUG and step width. Besides, Tai Chi was advantageous over brisk walking in improving BBS and step width. The improved BBS was correlated with enhanced visual network function and downregulation of interleukin-1ß. The improvements in UPDRS were associated with enhanced default mode network function, decreased L-malic acid and 3-phosphoglyceric acid, and increased adenosine and HIP2 mRNA levels. In addition, arginine biosynthesis, urea cycle, tricarboxylic acid cycle and beta oxidation of very-long-chain fatty acids were also improved by Tai Chi training. CONCLUSIONS: Long-term Tai Chi training improves motor function, especially gait and balance, in PD. The underlying mechanisms may include enhanced brain network function, reduced inflammation, improved amino acid metabolism, energy metabolism and neurotransmitter metabolism, and decreased vulnerability to dopaminergic degeneration. Trial registration This study has been registered at Chinese Clinical Trial Registry (Registration number: ChiCTR2000036036; Registration date: August 22, 2020).

Protective effects of polysaccharides from Atractylodes macrocephalae Koidz. against dextran sulfate sodium induced intestinal mucosal injury on mice.

In the present research, the water-soluble polysaccharides (AMP) from Atractylodes macrocephalae Koidz. were isolated and prepared. The protective effects of AMP on intestinal mucosal barrier injury induced by dextran sulfate sodium (DSS) in mice were investigated. It was found that AMP treatment significantly alleviated the body weight decreases and shorten colon length, and ameliorated colonic damage induced by DSS. Importantly, AMP prevented the over-expression of proinflammatory cytokines TNF-α, IL-1ß and IL-6, and decreased the infiltration of neutrophils in colon. Additionally, AMP could raise expressions of Mucin 2 and tight junction protein Claudin-1. AMP also modulated the intestinal microbiota by enhancing the overall richness and diversity, greatly reducing the proportion of harmful bacteria, for instance, Clostridiumsensu stricto1 and Escherichia Shigella, however, augmenting the ratio of potential beneficial bacteria such as Faecalibaculum and Bifidobacterium. This work offers some important insights on protective effects of polysaccharides AMP against intestinal barrier dysfunction and provides underlying mechanism of health-beneficial properties of these biological macromolecules.

Effects of dietary tryptophan on muscle growth, protein synthesis and antioxidant capacity in hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂.

The present study evaluated effects of dietary supplementation with tryptophan (Trp) on muscle growth, protein synthesis and antioxidant capacity in hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂. Fish were fed six different diets containing 2·6 (control), 3·1, 3·7, 4·2, 4·7 and 5·6 g Trp/kg diet for 56 d, respectively. Results showed that dietary Trp significantly (1) improved muscle protein content, fibre density and frequency of fibre diameter; (2) up-regulated the mRNA levels of PCNA, myf5, MyoD1, MyoG, MRF4, IGF-I, IGF-II, IGF-IR, PIK3Ca, TOR, 4EBP1 and S6K1; (3) increased phosphorylation levels of AKT, TOR and S6K1; (4) decreased contents of MDA and PC, and increased activities of CAT, GST, GR, ASA and AHR; (5) up-regulated mRNA levels of CuZnSOD, CAT, GST, GPx, GCLC and Nrf2, and decreased Keap1 mRNA level; (6) increased nuclear Nrf2 protein level and the intranuclear antioxidant response element-binding ability, and reduced Keap1 protein level. These results indicated that dietary Trp improved muscle growth, protein synthesis as well as antioxidant capacity, which might be partly related to myogenic regulatory factors, IGF/PIK3Ca/AKT/TOR and Keap1/Nrf2 signalling pathways. Finally, based on the quadratic regression analysis of muscle protein and MDA contents, the optimal Trp requirements of hybrid catfish (21·82-39·64 g) were estimated to be 3·94 and 3·93 g Trp/kg diet (9·57 and 9·54 g/kg of dietary protein), respectively.

Lentinan Combined with (125)I Brachytherapy for Recurrent Ovarian Cancer.

OBJECTIVE: To investigate the clinical value of lentinan combined with (125)I brachytherapy in the treatment of recurrent ovarian cancer. METHODS: A total of 160 patients with recurrent ovarian cancer admitted at Jiaozhou Central Hospital from June 2009 to October 2015 were enrolled in this study and randomly divided into observation group (80 cases) and control group (80 cases). The control group received chemotherapy. Observation group (80 cases) was treated with lentinan combined with (125)I brachytherapy on the basis of control group, and the efficacy, adverse reactions, and Karnofsky Performance Scale (KPS) and quality of life scale (QOL) scores of the two groups were analyzed and compared. RESULTS: After treatment, the levels of CA125, CA199, and CA724 in the 2 groups were markedly lower than those before treatment, and the observation group was lower than the control group (P < 0.05). After treatment, the proportion of CD4+/CD8+ cells and helper T cells and NK cells in the control group remarkably depleted, while the proportion of CD4+/CD8+ cells, NK cells, and B cells in the observation group increased significantly compared to that before treatment, and the level of IgA, IgG, and IgM in the control group decreased, while that in the observation group showed no conspicuous difference compared with that before chemotherapy (P > 0.05). The effective rate of observation group (85%) was higher than that of control group (75%) (P < 0.05). The overall survival of patients in the control group was (16.2 ± 2.04) months and that of the observation group was (24.8 ± 1.8) months. KPS and QOL scores in both groups were enormously higher than those before treatment, and the observation group was higher than the control group (P < 0.05). The incidence of hemoglobin reduction, leukopenia, aglobulia, granulocytopenia, nausea and vomiting, hepatorenal toxicity, and neurovirulence in the observation group was significantly lower than that in the control group. CONCLUSION: Lentinan combined with (125)I brachytherapy is effective in treating recurrent ovarian cancer, with mild adverse reactions and good tolerance.

Corydalis Rhizoma as a model for herb-derived trace metabolites exploration: A cross-mapping strategy involving multiple doses and samples.

Deciphering the metabolites of multiple components in herbal medicine has far-reaching significance for revealing pharmacodynamic ingredients. However, most chemical components of herbal medicine are secondary metabolites with low content whose in vivo metabolites are close to trace amounts, making it difficult to achieve comprehensive detection and identification. In this paper, an efficient strategy was proposed: herb-derived metabolites were predicted according to the structural characteristics and metabolic reactions of chemical constituents in Corydalis Rhizoma and chemical structure screening tables for metabolites were conducted. The fragmentation patterns were summarized from representative standards combining with specific cleavage behaviors to deduce structures of metabolites. Ion abundance plays an important role in compound identification, and high ion abundance can improve identification accuracy. The types of metabolites in different biological samples were very similar, but their ion abundance might be different. Therefore, for trace metabolites in biological samples, we used the following two methods to process: metabolites of high dose herbal extract were analyzed to characterize those of clinical dose herbal extracts in the same biological samples; cross-mapping of different biological samples was applied to identify trace metabolites based on the fact that a metabolite has different ion abundance in different biological samples. Compared with not using this strategy, 44 more metabolites of clinical dose herbal extract were detected. This study improved the depth, breadth, and accuracy of current methods for herb-derived metabolites characterization.

Synthesis of one-for-all type Cu5FeS4 nanocrystals with improved near infrared photothermal and Fenton effects for simultaneous imaging and therapy of tumor.

CuS materials exhibit excellent near infrared (NIR) photoabsorption and photothermal effect, but they are lack of magnetic resonance imaging (MRI) ability. Fe-based nanomaterials possess MRI capacity, but they usually exhibit poor NIR photoabsorption. In order to solve the above problems, we synthesize three kinds of CuxFeySz samples, including FeS2, CuFeS2 and Cu5FeS4 nanomaterials. With the Cu/Fe ratios increase from 0/1.0 to 1.0/1.0 and 5.0/1.0, the localized surface plasmon resonances (LSPRs) characteristic peaks shift to longer wavelength, and the photothermal transduction efficiencies go up from 24.4% to 36.6% and 45.9%. Thus, Cu5FeS4 is found to be the most excellent sample. Especially, Cu5FeS4 exhibits photothermal-enhanced Fenton effect, which can produce hydroxyl radical (·OH) under a wide pH range (e.g., pH = 5.4-7.4) to realize the chemodynamic effect. In addition, Cu5FeS4 can be employed as an efficient MRI contrast agent. When Cu5FeS4 dispersion is intravenously injected into the mouse, the tumor can be detected by MRI as well as thermal imaging, and eliminated through photothermal-enhanced chemodynamic effect. Therefore, Cu5FeS4 can be used as an efficient "one-for-all" type agent for MRI-guided photothermal-enhanced chemodynamic therapy of tumor.

Cell membrane camouflaged bismuth nanoparticles for targeted photothermal therapy of homotypic tumors.

Bi nanoparticles (NPs) have been demonstrated as effective all-in-one type theranostic agent for imaging-guided photothermal therapy, but their applications have been limited by relatively low biocompatibility and target accumulation capacity. To address this issue, we report the camouflage of Bi NPs (size: ~42 ± 2 nm) by using the mouse colon cancer CT26 cells membrane (CT26 CCM). The camouflaging process confers the efficient coating of CCM shell layer with thickness of ~8 ± 2 nm on Bi NPs cores, which can be confirmed by TEM image, zeta potential and protein gel electrophoresis tests. Simultaneously, CCM shell has no side effects on the photoabsorption/photothermal effect. Importantly, Bi@CCM NPs retain significant features of CCM, including good biocompatibility and homologous targeting ability. When Bi@CCM dispersion was intravenously (i.v.) injected into mice, they exhibited higher blood circulation half-life (11.5 h, ~2.9 times) and accumulation amount (4.7 ± 0.56% ID/g, ~2.3 times) in homotypic CT26 tumor compared to those (4.0 h in blood and 2.03 ± 0.60% ID/g in tumor) from uncoated Bi NPs. After 808 nm laser irradiation, CT26 cancer cells could be effectively ablated after the photothermal therapy of high-accumulated Bi@CCM NPs, and then the tumor tends to be eradicated after 12 days. Thus, Bi NPs camouflaged with CT26 CCM have great potential for the targeted photothermal therapy of homotypic tumors.

Isoleucine improved growth performance, and intestinal immunological and physical barrier function of hybrid catfish Pelteobagrus vachelli × Leiocassis longirostris.

This study was performed to determine effects of dietary isoleucine (Ile) on growth performance, and intestinal immunological and physical barrier function of hybrid catfish Pelteobagrus vachelli × Leiocassis longirostris. Six hundred and thirty fish (33.11 ± 0.09 g) were randomly divided into seven experimental groups with three replicates each, and respectively fed seven diets with 5.0, 7.5, 10.0, 12.5, 15.0, 17.5, and 20.0 g Ile kg-1 diets for 8 weeks. The results showed improvement of growth performance, feed intake, feed utilization, relative gut length (RGL), and intestinal fold height and width by dietary Ile (P < 0.05). Meanwhile, dietary Ile (12.5 g kg-1 diet) improved the activities of lysozyme (LZM), acid phosphatase, alkaline phosphatase and the contents of complement 3 (C3), C4, and immunoglobulin M (IgM) (P < 0.05). The c-type-lectin, c-LZM, g-LZM, and hepcidin mRNA expressions in the intestine were up-regulated in fish fed diets with 10.0-20.0 g Ile kg-1 diet (P < 0.05). Dietary Ile (10.0-12.5 g Ile kg-1 diet) increased intestinal ß-defensin mRNA expression partially in association with Sirt1/ERK/90RSK signaling pathway. Dietary Ile (12.5-15.0 g Ile kg-1 diet) decreased oxidative damage and improved antioxidant ability by increasing activities and expressions of superoxide dismutase, glutathione peroxidase, and glutathione reductase, glutathione-S-transferase (P < 0.05). The occludin, ZO-1, ZO-2, claudin3, and claudin 7 mRNA expressions in the intestine were up-regulated in fish fed diets with 10.0 and 12.5 g Ile kg-1 diet (P < 0.05), whereas the myosin light chain kinase gene expression was decreased in fish fed diets with 7.5-17.5 g Ile kg-1 diet. Dietary Ile (10-12.5 g Ile kg-1 diet) decreased apoptotic responses by reducing the expression of caspase3 and caspase 9 via the AKT/TOR signaling pathway. Based on the quadratic regression analysis of PWG, the dietary Ile requirement of hybrid catfish was estimated to be 12.43 g Ile kg-1 diet, corresponding to 32.05 g Ile kg-1 dietary protein. Collectively, dietary Ile improved growth performance and immunological and physical barrier function of intestine in hybrid catfish.

Corydalis Rhizoma as a model for herb-derived trace metabolites exploration:A cross-mapping strategy involving multiple doses and samples / 药物分析学报

Deciphering the metabolites of multiple components in herbal medicine has far-reaching significance for revealing pharmacodynamic ingredients.However,most chemical components of herbal medicine are secondary metabolites with low content whose in vivo metabolites are close to trace amounts,making it difficult to achieve comprehensive detection and identification.In this paper,an efficient strategy was proposed:herb-derived metabolites were predicted according to the structural characteristics and metabolic reactions of chemical constituents in Corydalis Rhizoma and chemical structure screening tables for metabolites were conducted.The fragmentation patterns were summarized from represen-tative standards combining with specific cleavage behaviors to deduce structures of metabolites.Ion abundance plays an important role in compound identification,and high ion abundance can improve identification accuracy.The types of metabolites in different biological samples were very similar,but their ion abundance might be different.Therefore,for trace metabolites in biological samples,we used the following two methods to process:metabolites of high dose herbal extract were analyzed to char-acterize those of clinical dose herbal extracts in the same biological samples;cross-mapping of different biological samples was applied to identify trace metabolites based on the fact that a metabolite has different ion abundance in different biological samples.Compared with not using this strategy,44 more metabolites of clinical dose herbal extract were detected.This study improved the depth,breadth,and accuracy of current methods for herb-derived metabolites characterization.

Osthole: A Traditional Chinese Medicine for Ocular Anti-Angiogenic Therapy.

PURPOSE: Osthole is an agent isolated from Cnidium monnieri (L.) Cusson and has been used to treat several disorders. Corneal neovascularization is a sight-threatening condition associated with several inflammatory or infectious ocular disorders. In this study, we investigated the anti-angiogenic effects of osthole on corneal neovascularization and the underlying mechanism. METHODS: MTT assay, HE staining, and calcein-AM/propidium iodide staining was conducted to detect the toxicity of osthole in vitro and in vivo. Corneal neovascularization of ICR mice was induced by alkali burn and observed by a slit lamp microscopy on day 7 after alkali injury. EdU assay, Ki67 immunofluorescence assay, Transwell migration assay, and Matrigel assay were conducted to investigate the role of osthole in endothelial angiogenic effects in vitro. Western blots were conducted to investigate the anti-angiogenic mechanism of osthole in corneal neovascularization. RESULTS: Administration of osthole ranging from 0.05 to 25 µM had no detectable cytotoxicity or tissue toxicity in vivo and in vitro. Topical administration of osthole inhibited corneal neovascularization induced by alkali burn. Osthole decreased the proliferation, migration, and tube-formation of endothelial cells induced by VEGF. Osthole inhibited endothelial angiogenic functions through blocking the phosphorylation of ERK1/2, JNK, and p38. CONCLUSION: Our study provides evidence that osthole is a promising drug for the treatment of corneal neovascularization.

Effect of dietary threonine on growth performance and muscle growth, protein synthesis and antioxidant-related signalling pathways of hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂.

The experiment was conducted to investigate the effects of dietary threonine (Thr) on growth performance and muscle growth, protein synthesis and antioxidant-related signalling pathways of hybrid catfish Pelteobagrus vachelli♀ × Leiocassis longirostris♂. A total of 1200 fish (14·19 (se 0·13) g) were randomly distributed into six groups with four replicates each, fed six diets with graded level of Thr (9·5, 11·5, 13·5, 15·4, 17·4 and 19·3 g/kg diets) for 56 d. Results showed (P < 0·05) that dietary Thr (1) increased percentage weight gain, specific growth rate, feed efficiency and protein efficiency ratio; (2) up-regulated growth hormone (GH), insulin-like growth factor 1 (IGF-1), proliferating cell nuclear antigen, myogenic regulation factors (MyoD, Myf5, MyoG and Mrf4) and myosin heavy chain (MyHC) mRNA levels; (3) increased muscle protein content via regulating the protein kinase B/target of rapamycin signalling pathway and (4) decreased malondialdehyde and protein carbonyl contents, increased catalase, glutathione-S-transferase, glutathione reductase and GSH activities, up-regulated mRNA levels of antioxidant enzymes related to NFE2-related factor 2 and γ-glutamylcysteine ligase catalytic subunit. These results suggest that Thr has a potential role to improve muscle growth and protein synthesis, which might be due to the regulation of GH-IGF system, muscle growth-related gene, antioxidative capacity and protein synthesis-related signalling pathways. Based on the quadratic regression analysis of specific growth rate, the Thr requirement of hybrid catfish (14·19-25·77 g) was estimated to be 13·77 g/kg of the diet (33·40 g/kg of dietary protein).

Vitamin D Alleviates Rotavirus Infection through a Microrna-155-5p Mediated Regulation of the TBK1/IRF3 Signaling Pathway In Vivo and In Vitro.

(1) Background: Vitamin D (VD) plays a vital role in anti-viral innate immunity. However, the role of VD in anti-rotavirus and its mechanism is still unclear. The present study was performed to investigate whether VD alleviates rotavirus (RV) infection through a microRNA-155-5p (miR-155-5p)-mediated regulation of TANK-binding kinase 1 (TBK1)/interferon regulatory factors 3 (IRF3) signaling pathway in vivo and in vitro. (2) Methods: The efficacy of VD treatment was evaluated in DLY pig and IPEC-J2. Dual-luciferase reporter activity assay was performed to verify the role of miR-155-5p in 1α,25-dihydroxy-VD3 (1,25D3) mediating the regulation of the TBK1/IRF3 signaling pathway. (3) Results: A 5000 IU·kg-1 dietary VD3 supplementation attenuated RV-induced the decrease of the villus height and crypt depth (p < 0.05), and up-regulated TBK1, IRF3, and IFN-ß mRNA expressions in the jejunum (p < 0.05). Incubation with 1,25D3 significantly decreased the RV mRNA expression and the RV antigen concentration, and increased the TBK1 mRNA and protein levels, and the phosphoprotein IRF3 (p-IRF3) level (p < 0.05). The expression of miR-155-5p was up-regulated in response to an RV infection in vivo and in vitro (p < 0.05). 1,25D3 significantly repressed the up-regulation of miR-155-5p in vivo and in vitro (p < 0.05). Overexpression of miR-155-5p remarkably suppressed the mRNA and protein levels of TBK1 and p-IRF3 (p < 0.01), while the inhibition of miR-155-5p had an opposite effect. Luciferase activity assays confirmed that miR-155-5p regulated RV replication by directly targeting TBK1, and miR-155-5p suppressed the TBK1 protein level (p < 0.01). (4) Conclusions: These results indicate that miR-155-5p is involved in 1,25D3 mediating the regulation of the TBK1/IRF3 signaling pathway by directly targeting TBK1.

The underlying mechanisms of Jie-Du-Hua-Yu granule for protecting rat liver failure.

OBJECTIVES: Jie-Du-Hua-Yu (JDHY) granule is a combination of six traditional Chinese medicines with known therapeutic effect in treating acute liver failure (ALF). The aim of this study was to investigate the amelioration efficacy of JDHY in lipopolysaccharide/D-galactosamine (LPS/D-GalN)-induced ALF in rat and explore the possible molecular mechanism underlying the therapeutic efficacy. MATERIALS AND METHODS: The efficacy of JDHY was determined by assessing hepatic pathology and function in LPS and D-GalN challenged Wistar rat. We also evaluated the effect of JDHY on LPS-induced Kupffer cells by measuring inflammatory cytokines and determining the phenotypic function. By means of bioinformatics analysis of liver tissue and validation in Kupffer cells, we identified possible pathways involved in the pharmacologic action of mechanism of JDHY. RESULTS: JDHY could attenuate LPS-induced liver injury in rat by inhibiting apoptosis and increasing hepatic activity. In vitro study showed that JDHY could decrease the production of proinflammatory cytokines (tumor necrosis factor-α, IL6, and interferon-γ), increase anti-inflammatory cytokines (IL10, IL13), and promote cell survival and proliferation, possibly due to inhibition of IκB/nuclear factor-κB (NF-κB) signaling pathway and expression of CD14 and CXCL2, which was consistent with the findings from bioinformatics analysis. CONCLUSION: Our results revealed that JDHY protected against LPS-induced liver damage both in vitro and in vivo, by inhibiting the NF-κB-mediated inflammatory pathway, indicating its potential function to treat liver diseases.