Metabolic systems approaches update molecular insights of clinical phenotypes and cardiovascular risk in patients with homozygous familial hypercholesterolemia.

BACKGROUND: Homozygous familial hypercholesterolemia (HoFH) is an orphan metabolic disease characterized by extremely elevated low-density lipoprotein cholesterol (LDL-C), xanthomas, aortic stenosis, and premature atherosclerotic cardiovascular disease (ASCVD). In addition to LDL-C, studies in experimental models and small clinical populations have suggested that other types of metabolic molecules might also be risk factors responsible for cardiovascular complications in HoFH, but definitive evidence from large-scale human studies is still lacking. Herein, we aimed to comprehensively characterize the metabolic features and risk factors of human HoFH by using metabolic systems strategies. METHODS: Two independent multi-center cohorts with a total of 868 individuals were included in the cross-sectional study. First, comprehensive serum metabolome/lipidome-wide analyses were employed to identify the metabolomic patterns for differentiating HoFH patients (n = 184) from heterozygous FH (HeFH, n = 376) and non-FH (n = 100) subjects in the discovery cohort. Then, the metabolomic patterns were verified in the validation cohort with 48 HoFH patients, 110 HeFH patients, and 50 non-FH individuals. Subsequently, correlation/regression analyses were performed to investigate the associations of clinical/metabolic alterations with typical phenotypes of HoFH. In the prospective study, a total of 84 HoFH patients with available follow-up were enrolled from the discovery cohort. Targeted metabolomics, deep proteomics, and random forest approaches were performed to investigate the ASCVD-associated biomarkers in HoFH patients. RESULTS: Beyond LDL-C, various bioactive metabolites in multiple pathways were discovered and validated for differentiating HoFH from HoFH and non-FH. Our results demonstrated that the inflammation and oxidative stress-related metabolites in the pathways of arachidonic acid and lipoprotein(a) metabolism were independently associated with the prevalence of corneal arcus, xanthomas, and supravalvular/valvular aortic stenosis in HoFH patients. Our results also identified a small marker panel consisting of high-density lipoprotein cholesterol, lipoprotein(a), apolipoprotein A1, and eight proinflammatory and proatherogenic metabolites in the pathways of arachidonic acid, phospholipid, carnitine, and sphingolipid metabolism that exhibited significant performances on predicting first ASCVD events in HoFH patients. CONCLUSIONS: Our findings demonstrate that human HoFH is associated with a variety of metabolic abnormalities and is more complex than previously known. Furthermore, this study provides additional metabolic alterations that hold promise as residual risk factors in HoFH population.

Integrated analysis of microRNA and mRNA expression profiles in homozygous familial hypercholesterolemia patients and validation of atherosclerosis associated critical regulatory network.

Homozygous familial hypercholesterolemia (HoFH) is a rare, life-threatening genetic disorder characterized by an extremely elevated serum level of low-density lipoprotein cholesterol (LDL-C) and accelerated premature atherosclerotic cardiovascular diseases (ASCVD). However, the detailed mechanism of how the pathogenic mutations of HoFH trigger the acceleration of ASCVD is not well understood. Therefore, we performed high-throughput RNA and small RNA sequencing on the peripheral blood RNA samples of six HoFH patients and three healthy controls. The gene and miRNA expression differences were analyzed, and seven miRNAs and six corresponding genes were screened out through regulatory network analysis. Validation through quantitative PCR of genes and miRNAs from 52 HoFH patients and 20 healthy controls revealed that the expression levels of hsa-miR-486-3p, hsa-miR-941, and BIRC5 were significantly upregulated in HoFH, while ID1, PLA2G4C, and CACNA2D2 were downregulated. Spearman correlation analysis found that the levels of ID1, hsa-miR-941, and hsa-miR-486-3p were significantly correlated with additional ASCVD risk factors in HoFH patients. This study represents the first integrated analysis of transcriptome and miRNA expression profiles in patients with HoFH, a rare disease, and as a result, six differentially expressed miRNAs/genes that may be related to atherosclerosis in HoFH are reported. The miRNA-mRNA regulatory network may be the critical regulation mechanism by which ASCVD is accelerated in HoFH.

Prognostic value of coronary flow velocity reserve in patients with homozygous familial hypercholesterolemia.

BACKGROUND: Coronary flow velocity reserve (CFVR) can provide useful quantitative information on the functional status of coronary artery circulation, and an impaired CFVR (< 2.0) was associated with a significant increase in the occurrence of cardiac events. Coronary artery disease (CAD) is the leading cause of death in homozygous familial hypercholesterolemia (HoFH), but the relationship between impaired CFVR and outcome in HoFH has never been discussed before METHODS: To explore the long-term prognostic value of CFVR in patients with HoFH, 39 HoFH patients with CFVR data (mean age with 16.7 years) were enrolled from the Genetic and Imaging of Familial Hypercholesterolemia in Han Nationality Study. All patients were divided into impaired CFVR (CFVR < 2.0, n = 17) and preserved CFVR (CFVR≥2.0, n = 22) group. Follow-up was performed until a major adverse cardiac event (MACE) occurred or up to June 30, 2020 RESULTS: During a median follow-up of 89 months, 16 events were registered, 12 of which were occurred in the impaired CFVR group and four occurred in the preserved CFVR group. The event-free survival rate of impaired CFVR group was significantly lower than that in the preserved CFVR group (29.4% vs 81.8%, P < .001), and CFVR < 2.0 was independently associated with prognosis before and after adjustment for related risk factors (HR 5.197, 95% CI 1.669 to 16.178, P = .004 and HR 5.488, 95% CI 1.470 to 20.496, P = .011, respectively) CONCLUSIONS: an impaired CFVR predicts a worse outcome in HoFH. CFVR shows an independent value in the prediction of long-term outcome in HoFH.

Effect of Pyrolysis Temperature on the Characterisation of Dissolved Organic Matter from Pyroligneous Acid.

Dissolved organic matter (DOM) greatly influences the transformation of nutrients and pollutants in the environment. To investigate the effects of pyrolysis temperatures on the composition and evolution of pyroligneous acid (PA)-derived DOM, DOM solutions extracted from a series of PA derived from eucalyptus at five pyrolysis temperature ranges (240-420 °C) were analysed with Fourier transform infrared spectroscopy, gas chromatography-mass spectroscopy, and fluorescence spectroscopy. Results showed that the dissolved organic carbon content sharply increased (p < 0.05) with an increase in pyrolysis temperature. Analysis of the dissolved organic matter composition showed that humic-acid-like substances (71.34-100%) dominated and other fluorescent components (i.e., fulvic-acid-like, soluble microbial by-products, and proteinlike substances) disappeared at high temperatures (>370 °C). The results of two-dimensional correlation spectroscopic analysis suggested that with increasing pyrolysis temperatures, the humic-acid-like substances became more sensitive than other fluorescent components. This study provides valuable information on the characteristic evolution of PA-derived DOM.

Comparison of long-term outcomes of young patients after a coronary event associated with familial hypercholesterolemia.

OBJECTIVE: Familial hypercholesterolemia (FH) is an important cause of premature coronary artery disease (CAD). Prognosis data are lacking in patients with FH and coronary artery disease particularly in the era of widespread statin use. We compared long-term prognosis between patients with and without FH after a coronary event. METHODS: In this retrospective study, 865 patients younger than 40 years of age with CAD were enrolled. FH was diagnosed based on the Dutch Lipid Clinic Network algorithm. Baseline characteristics, coronary angiographic findings and prognosis during median follow-up of 5 (3-8) years were compared between patients with or without FH. RESULTS: Definite or probable FH was detected in 37 patients (4.3%) and possible FH in 259 patients (29.9%). FH was associated with significantly higher prevalence of multi-vessel lesions (p < 0.001) and higher Gensini score (p = 0.008). In the subset of 706 patients for whom follow-up data were available, 127 (18.0%) suffered major adverse cardiovascular and cerebrovascular events (MACCE). FH was associated with increased risk of MACCE, independently of age, sex, smoking, body mass index, hypertension or diabetes mellitus (HR = 2.30, 95%CI = 1.09 to 4.84, p = 0.028). CONCLUSIONS: FH is an independent risk factor for MACCE in young patients after a coronary event during long-term follow-up. It is necessary to optimize lipid treatment of patients with FH after a coronary event.

Recombinant HDL (Milano) protects endotoxin-challenged rats from multiple organ injury and dysfunction.

Endotoxemia, the systemic inflammatory host response to infection, leads to severe septic shock and multiple organ injury and dysfunction syndrome (MOPS), which cause mortality. Apolipoprotein A-IMilano (apoAIM), a naturally occurring cysteine mutant of apoAI with dimers as its effective form, showed an enhanced cardiovascular protective activity compared with wild-type apoAI (apoAIwt). To investigate the role of recombinant high-density lipoprotein (rHDL) reconstituted with apoAIM (rHDLM) on endotoxemia and MOPS, we examined the anti-inflammatory, anti-oxidant, and protective effects of this cysteine mutant against organ injury in endotoxin-challenged rat models compared with rHDLwt. In the present study, we demonstrated for the first time that pretreatment with rHDLM significantly attenuated liver and renal dysfunction and histopathological features of lung injury in endotoxin-challenged endotoxemia rats. Administration of rHDLM to endotoxemia rats dramatically suppressed proinflammatory cytokines and adhesion molecule increase in tumor necrosis factor α, interleukin 1ß, interleukin 6, and intercellular adhesion molecule 1. In addition, rHDLM pretreatment inhibited lipid peroxidation and enhanced total antioxidant capacity in vivo. In comparison with rHDLwt, rHDLM showed enhanced capacity on anti-inflammatory and anti-oxidant functions. In summary, administration of rHDLM protected endotoxin-challenged endotoxemia and MOPS through enhanced anti-inflammatory and anti-oxidant properties.

Interleukin-6/signal transducer and activator of transcription 3 (STAT3) pathway is essential for macrophage infiltration and myoblast proliferation during muscle regeneration.

Inflammation and microenvironment play a crucial role in muscle regeneration. IL (interleukin)-6, as a multifunctional cytokine is involved in the processes. However, the causative effect of IL-6 in muscle regeneration remains unclear. In a mouse model of cardiotoxin-induced muscle injury/regeneration, infiltrated monocytes/macrophages produce a high level of IL-6 started on 1 day (24 h) after injury. In IL-6 knock-out (-/-) mice, the muscle regeneration procedure was impaired along with decreased myogenic determination factor (MyoD) and myogenin mRNA level and increased interstitial fibrosis. The IL-6(-/-) mice exhibited less macrophage infiltration, lower inflammatory cytokine (IL-1ß, inducible NO synthase, Transforming growth factor (TGF)-ß1, and IL-10) and chemokine (CCL2, CCL3, and CCL5) expression, and inhibited myoblast proliferation. In vitro, IL-6 deficiency or Signal Transducer and Activator of Transcription 3 (STAT3) knockdown in activated macrophage attenuated the expression of CCL2, CCL3, but not CCL5, which resulted in less macrophage migration. Moreover, inflammatory macrophages promoted myoblast proliferation in an IL-6-dependent manner. Finally, adoptive transfer IL-6(+/+) BM cells into IL-6(-/-) mice rescued the impaired regeneration with improved MyoD and myogenin expression. Taken together, IL-6 expression and the activated STAT3 signaling pathway in monocytes/macrophages is a critical mediator of macrophage migration and myoblast proliferation during muscle regeneration.

[Involvement of chemokine CXCL16 in myocardial infarction and its influence on phagocytic activity of macrophage in vitro].

OBJECTIVE: To explore whether chemokine CXCL16 is up-regulated after myocardial infarction and promotes the phagocytic activity of macrophage in vitro. METHODS: Forty wild-type mice were randomly separated into 2 groups (n = 20 each). Group A had the ligation of left anterior descending coronary artery while group B underwent a sham operation. Electrocardiogram was used to assess whether the operation was successful or not. Three days after surgery, 10 animals of each group were sacrificed and the serum level of CXCL16 was detected by enzyme-linked immunosorbent assay (ELISA). Twenty-eight days after surgery, cardiac function of the remaining mice was measured by small animal ultrasound. Then the animals were sacrificed. Hematoxylin and eosin (HE) staining and immunohistochemical staining of cardiac paraffin section were used to observe the inflammation and detect the expression of CXCL16 in cardiac tissue after myocardial infarction. To explore the function of CXCL16 in vitro, primary murine monocytes were separated from bone marrow, cultured to differentiate into macrophages and transfected with adenovirus vectors over-expressing CXCL16 or control adenovirus vectors. After stimulation by debris of cardiac cells, the phagocytic uptake by macrophages was evaluated by flow cytometry. RESULTS: The model of myocardial infarction was successfully established. ELISA showed that the serum level of CXCL16 was elevated 3 days after myocardial infarction [(1 079 ± 176) vs (611 ± 37) pg/ml, P = 0.032]. HE and immunohistochemical staining demonstrated that the infiltration of macrophages increased during an early stage of myocardial infarction and decreased at the late stage. The CXCL16 level was up-regulated 3 days after myocardial infarction and returned to normal level at Day 28. Furthermore, macrophages transfected with adenovirus over-expressing CXCL16 showed stronger phagocytic activity compared with control (17.11% ± 0.87% vs 7.91% ± 0.71%, P < 0.01). CONCLUSION: CXCL16 is up-regulated after myocardial infarction in mice. And an in vitro over-expression of CXCL16 promotes the macrophage phagocytosis of cardiac debris.

Rhodiosin from Rhodiola crenulata effectively alleviate postprandial hyperglycemia by inhibiting both the activity and production of α­glucosidase.

BACKGROUND: Effective control of postprandial blood glucose (PBG) level is essential for the prevention and treatment of diabetes and its complications. Several flavonoids have attracted much attention due to their significant PBG-lowering effects. However, there is still a certain gap in the in vivo hypoglycemic activity of most flavonoids compared to first-line drugs available on the market, and are still lack of the PBG-lowering effects of 8-hydroxyflavones and their structure-activity relationship. PURPOSE: Evaluate hypoglycemic effects of 8-hydroxyflavones from Rhodiola crenulata in vitro and in vivo, especially comparatively analyze the relationship between hypoglycemic effects and flavonoid configuration and reveal the possible mechanism of 8-hydroxyflavones in lowering hyperglycemia. METHODS: Starch, maltose, sucrose, and glucose tolerance tests in both diabetic and normal mice were used to evaluate and compare the hypoglycemic effects of 8-hydroxyflavones rhodiosin (RHS), rhodionin (RHN), and herbacetin (HBT). Molecular docking, enzyme kinetics, and immunofluorescence analysis were used to research the possible hypoglycemic mechanisms of 8-hydroxyflavones. RESULTS: RHS (5 and 10 mg/kg) could efficiently decrease PBG levels in both normal and diabetes mice. Moreover, RHS, RHN, and HBT all had significant PBG-lowering effects in transgenic diabetes mice, and the effects were equivalent to or stronger than acarbose. Further mechanism research indicated that 8-hydroxyflavones achieved PBG-lowering effects by inhibiting both the activity and production of glycosidase. Notably, we have innovatively discovered that inhibiting the expression of glycosidases rather than just their activities may be a new target for hypoglycemic drugs. CONCLUSION: We have firstly comprehensively and systematically clarified PBG-lowering effects of 8-hydroxyflavones from Rhodiola crenulata, and revealed their structure-activity relationships and hypoglycemic mechanisms. The study demonstrated that the substitution of 8-hydroxy groups in flavonoids could significantly enhance their hypoglycemic effects, which were equivalent to or stronger than commercially available drug acarbose. 8-Hydroxyflavones could be used as therapeutic or health drugs with significant potential to reduce postprandial hyperglycemia.

Differences in the uptake and translocation of differentially charged microplastics by the taproot and lateral root of mangroves.

The interception of microplastics (MPs) by mangrove roots plays an indispensable role in reducing the environmental risks of MPs. However, there remains limited research on the fate of the intercepted MPs. Hereby, the uptake and subsequent translocation of 0.2 µm and 2 µm PS MPs with different coating charge by the typical salt-secreting mangrove plants (Aegiceras corniculatum) were investigated. Compared to amino-functionalized PS with positive charge (PS-NH2), the visualized results indicated that the efficient uptake of carboxy-functionalized PS with negative charge (PS-COOH) was more dependent on taproots. But for the lateral roots, it only allowed the entry of PS-NH2 instead of PS-COOH. The specific uptake pathways of PS-NH2 on the lateral roots could attribute to the release of H+ and organic acids by root hairs, as well as the relative higher Zeta potential. After entering the Aegiceras corniculatum roots, the translocation of PS MPs was restricted by their particle sizes. Furthermore, the release of PS MPs from Aegiceras corniculatum leaf surfaces through the salt glands and stomata was observed. And the decline in the photochemical efficiency of leaves under PS MPs exposure also indirectly proved the foliar emission of PS MPs. Our study improved the understanding of the environmental behaviors and risks of the retained MPs in mangroves.

Health assessment based on exposure to microplastics in tropical agricultural soil.

Microplastic (MP) pollution of agricultural soils has caused global alarm over its widespread distribution and potential risks to terrestrial ecosystems and human health. This study assessed human health based on exposure to soil MPs through a comprehensive investigation of the factors influencing their occurrence and spatial distribution on Hainan Island, South China. The results showed that the abundance of soil MPs was 1128.6 ± 391.5 items·kg-1, whereas the normalized abundance of MPs based on using a power-law function was 19,261.4 items·kg-1. Regarding the extent of population exposure to agricultural soil MPs, the average daily exposure dose (pADD) model revealed that using mass as an indicator to assess the health risks associated with MP intake is more reliable than using abundance. However, abundance-based exposure assessments are also relevant because MPs with smaller particle sizes are more harmful to human health. Moreover, for adults, the normalized pADD values based on abundance and mass were 1.68E-02 item MPs·kg BW-1·d-1 and 7.23E-02 mg MPs·kg BW-1·d-1, respectively. Although the multidimensionality of MPs should be further aligned and quantified, the preliminary findings of this study contribute to the development of human health risk assessment frameworks for soil MPs.

Extract of Silphium perfoliatum L. improve lipid accumulation in NAFLD mice by regulating AMPK/FXR signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Globally, the incidence rate and number of patients with nonalcoholic fatty liver disease are increasing, which has become one of the greatest threats to human health. However, there is still no effective therapy and medicine so far. Silphium perfoliatum L. is a perennial herb native to North America, which is used to improve physical fitness and treat liver and spleen related diseases in the traditional medicinal herbs of Indian tribes. This herb is rich in chlorogenic acids, which have the functions of reducing blood lipids, losing weight and protecting liver. However, the effect of these compounds on nonalcoholic fatty liver disease remains unclear. AIM OF THE STUDY: Clarify the therapeutic effects and mechanism of the extract (CY-10) rich in chlorogenic acid and its analogues from Silphium perfoliatum L. on non-alcoholic fatty liver disease, and to determine the active compounds. MATERIALS AND METHODS: A free fatty acid-induced steatosis model of HepG2 cells was established to evaluate the in vitro activity of CY-10 in promoting lipid metabolism. Further, a high-fat diet-induced NAFLD model in C57BL/6 mice was established to detect the effects of CY-10 on various physiological and biochemical indexes in mice, and to elucidate the in vivo effects of the extract on regulating lipid metabolism, anti-inflammation and hepatoprotection, and nontarget lipid metabolomics was performed to analyze differential metabolites of fatty acids in the liver. Subsequently, western blotting and immunohistochemistry were used to analyze the target of the extract and elucidate its mechanism of action. Finally, the active compounds in CY-10 were elucidated through in vitro activity screening. RESULTS: The results indicated that CY-10 significantly attenuated lipid droplet deposition in HepG2 cells. The results of in vivo experiments showed that CY-10 significantly reduce HFD-induced mouse body weight and organ index, improve biochemical indexes, oxidation levels and inflammatory responses in the liver and serum, thereby protecting the liver tissue. It can promote the metabolism of unsaturated fatty acids in the liver and reduce the generation of saturated fatty acids. Furthermore, it is clarified that CY-10 can promote lipid metabolism balance by regulating AMPK/FXR/SREPB-1c/PPAR-γ signal pathway. Ultimately, the main active compound was proved to be cryptochlorogenic acid, which has a strong promoting effect on the metabolism of fatty acids in cells. Impressively, the activities of CY-10 and cryptochlorogenic acid were stronger than simvastatin in vitro and in vivo. CONCLUSION: For the first time, it is clarified that the extract rich in chlorogenic acids and its analogues in Silphium perfoliatum L. have good therapeutic effects on non-alcoholic fatty liver disease. It is confirmed that cryptochlorogenic acid is the main active compound and has good potential for medicine.

Characterization of alkaloids and phenolics in Nitraria roborowskii Kom. fruit by UHPLC-triple-TOF-MS/MS and its sucrase and maltase inhibitory effects.

Nitraria roborowskii Kom (NRK), with high economic and ecological value, is mainly distributed in the Qaidam Basin, China. However, research on its chemical components and bioactivities is still rare. In this study, its chemical constituents (52) including 10 ß-carboline alkaloids, nine cyclic peptides, three indole alkaloids, five pyrrole alkaloids, eight phenolic acids and 17 flavonoids were identified tentatively using UPLC-triple-TOF-MS/MS. Notablely, one new ß-carboline alkaloid and five new cyclic peptides were confirmed using MS/MS fragmentation pathways. In addition, experiments in vitro indicated that NRK-C had strong maltase and sucrase inhibitory activities (IC50 of 0.202 and 0.103 mg/mL, respectively). Polysaccharide tolerance experiments confirmed NRK-C (400 mg/kg) was associated with decreased postprandial blood glucose (PBG) in diabetic mice. These results suggested that NRK fruit might be used as a functional ingredient in food products.

Angiotension II directly bind P2X7 receptor to induce myocardial ferroptosis and remodeling by activating human antigen R.

Continuous remodeling of the heart can result in adverse events such as reduced myocardial function and heart failure. Available evidence indicates that ferroptosis is a key process in the emergence of cardiac disease. P2 family purinergic receptor P2X7 receptor (P2X7R) activation plays a crucial role in numerous aspects of cardiovascular disease. The aim of this study was to elucidate any potential interactions between P2X7R and ferroptosis in cardiac remodeling stimulated by angiotensin II (Ang II), and P2X7R knockout mice were utilized to explore the role of P2X7R and elucidate its underlying mechanism through molecular biological methods. Ferroptosis is involved in cardiac remodeling, and P2X7R deficiency significantly alleviates cardiac dysfunction, remodeling, and ferroptosis induced by Ang II. Mechanistically, Ang II interacts with P2X7R directly, and LYS-66 and MET-212 in the in the ATP binding pocket form a binding complex with Ang II. P2X7R blockade influences HuR-targeted GPX4 and HO-1 mRNA stability by affecting the shuttling of HuR from the nucleus to the cytoplasm and its expression. These results suggest that focusing on P2X7R could be a possible therapeutic approach for the management of hypertensive heart failure.

Pregnancy-associated plasma protein-A as a marker of culprit lesion instability in unstable angina patients: an intravascular ultrasound study.

BACKGROUND: We evaluated the relationship between pregnancy-associated plasma protein-A (PAPP-A) and coronary plaque instability as assessed by intravascular ultrasound (IVUS). METHODS: We performed greyscale IVUS analysis in culprit lesions of 93 patients with unstable angina (UA) and 72 with stable angina (SA). A sandwich enzyme-linked immunosorbent assay technique was used to assay circulating PAPP-A. RESULTS: Patients with UA had higher PAPP-A levels than those with SA 10.8 mIU/l [interquartile range (IQR) 8.3-14.4] vs. 5.4 (IQR 2.9-9.8) mIU/l, p < 0.001]. Lesions in patients with higher PAPP-A levels were associated with larger plaque burden than lesions in patients with lower PAPP-A levels. IVUS attenuated plaque, positive remodeling and plaque rupture. Thrombus and angiographic Ambrose type-II eccentric lesions or multiple irregularities were more common in patients with higher PAPP-A levels than in those with lower PAPP-A levels. They were also more common in patients with UA and higher PAPP-A levels than in patients with (1) SA and higher PAPP-A levels, (2) UA and lower PAPP-A levels or (3) SA and lower PAPP-A levels. CONCLUSIONS: Higher PAPP-A levels were associated with coronary plaque instability in vivo and unstable symptoms in patients with coronary artery disease.

An unrecognized entry pathway of submicrometre plastics into crop root: The split of hole in protective layer.

Threats to food safety caused by the continuous accumulation of plastic particles in the terrestrial environment is currently a worldwide concern. To date, descriptions of how plastic particles pass the external biological barrier of crop root have been vague. Here, we demonstrated that submicrometre polystyrene particles passed unimpededly the external biological barrier of maize through the split of holes in the protective layer. We identified plastic particles induced the apical epidermal cells of root tips become round, thereby expanding the intercellular space. It further pulled apart the protective layer between the epidermal cells, and eventually formed the entry pathway for plastic particles. The enhancement of oxidative stress level induced by plastic particles was the main reason for the deformation of the apical epidermal cells (increased roundness values: 15.5%), comparing to the control. Our findings further indicated that the presence of cadmium was conducive to the "holes" formation. Our results highlighted the critical insights into the fracture mechanisms of plastic particles for the external biological barriers of crop roots, providing a strong incentive to access the risk of plastic particles in agriculture security.

Metabolomic Approach to Screening Homozygotes in Chinese Patients with Severe Familial Hypercholesterolemia.

Homozygous familial hypercholesterolemia (HoFH) is a rare inborn-errors-of-metabolism disorder characterized by devastatingly elevated low-density lipoprotein cholesterol (LDL-C) and premature cardiovascular disease. The gold standard for screening and diagnosing HoFH is genetic testing. In China, it is expensive and is always recommended for the most likely HoFH subjects with aggressive LDL-C phenotype. However, the LDL-C levels of HoFH patients and a substantial proportion of heterozygous FH (HeFH) patients overlapped considerably. Here, we performed a cost-effective metabolomic profiling on genetically diagnosed HoFH (n = 69) and HeFH patients (n = 101) with overlapping LDL-C levels, aiming to discovery a unique metabolic pattern for screening homozygotes in patients with severe FH. We demonstrated a differential serum metabolome profile in HoFH patients compared to HeFH patients. Twenty-one metabolomic alterations showed independent capability in differentiating HoFH from severe HeFH. The combined model based on seven identified metabolites yielded a corrected diagnosis in 91.3% of HoFH cases with an area under the curve value of 0.939. Collectively, this study demonstrated that metabolomic profiling serves as a useful and economical approach to preselecting homozygotes in FH patients with severe hypercholesterolemia and may help clinicians to conduct selective genetic confirmation testing and familial cascade screening.

Enrichment of Wheat Bread with Platycodon grandiflorus Root (PGR) Flour: Rheological Properties and Microstructure of Dough and Physicochemical Characterization of Bread.

Platycodon grandiflorus (Jacq.) A.DC. root (PGR) flour is well known for its medical and edible values. In order to develop nutritionally fortified products, breads were prepared using wheat flour, partially replaced with PGR flour. The rheological properties and microstructure of dough and the physicochemical characterization of bread were investigated. Results showed that lower level of PGR addition (3 and 6 g/100 g) would improve the baking performance of breads, while the higher level of PGR addition (9 g/100 g) led to smaller specific volume (3.78 mL/g), increased hardness (7.5 ± 1.35 N), and unpalatable mouthfeel (21.8% of resilience and 92.6% of springiness) since its negative effect on the viscoelasticity and microstructure of dough. Moreover, sensory evaluation analysis also showed that the PGR3 and PGR6 breads exhibited a similar flavor to the control bread, but the 9 g/100 g addition of PGR provided bread with an unpleasant odor through its richer volatile components. As expected, the phenolic content and antioxidant capacity of bread increased significantly (p < 0.05) as PGR flour was added to the bread formulation. The total phenolic content (TPC) ranged from 14.23 to 22.36 g GAE/g; thus, DPPH• and ABTS•+ scavenging capacity increased from 10.44 and 10.06 µg Trolox/g to 14.69 and 15.12 µg Trolox/g, respectively. Therefore, our findings emphasized the feasibility of PGR flour partially replacing wheat flour in bread-making systems.

Hypoglycemic effect of Nitraria tangutorum fruit by inhibiting glycosidase and regulating IRS1/PI3K/AKT signalling pathway and its active ingredient identification by UPLC-MS.

The hypoglycemic effect of NTB-40 (40% ethanol fraction of Nitraria tangutorum fruit) in type I/II diabetic mice and its underlying mechanism and active ingredient structure were investigated. The postprandial blood glucose (PBG) lowering effect of NTB-40 treatment was confirmed by maltose, starch, and sucrose tolerance tests in alloxan-induced DM mice and sucrase and maltase inhibitory activities in vitro. More importantly, long-term dosing experiments in high-fat diet-STZ-induced diabetic mice further demonstrated that NTB-40 intervention could improve glycolipid metabolism disorder and insulin resistance (IR) by maintaining glucose homeostasis (FBG, OGTT, ITT, FINS, and HOMA-IR) and lipid homeostasis (TC, TG, HDL-C, LDL-C, and FFA), reducing inflammation (IL-6, IL-1ß, and TNF-α) and oxidative stress (SOD and MDA), ameliorating the liver's histological structural abnormalities, and modulating the IRS1/PI3K/AKT signaling pathway and downstream targets (FOXO1, GSK3ß, GLUT4) for decreasing hepatic gluconeogenesis and promoting glycogen synthesis and glucose uptake. All these results indicated that NTB-40 had an anti-diabetic effect by modulating the IRS1/PI3K/AKT signaling pathway and inhibiting α-glucosidase activity. Finally, the main chemical components of NTB-40, including phenolic acids, flavonoids, and alkaloids, were assigned by UPLC-Triple-TOF MS/MS.

Inhibitory activities and rules of plant gallotannins with different numbers of galloyl moieties on sucrase, maltase and α-amylase in vitro and in vivo.

BACKGROUND: α-Glucosidase inhibitors could effectively reduce postprandial blood glucose (PBG) levels and control the occurrence of complications of diabetes. Gallotannins (GTs) in plants have attracted much attention due to their significant α-glucosidase inhibitory activities in vitro. However, there is still a lack of systematic comparative studies to further elucidate inhibitory activities in vivo and in vitro of these compounds against α-glucosidase, especially for mammalian sucrase and maltase, and analyze their structure-activity relationship. PURPOSE: Determine the in vitro and in vivo inhibitory activities of five GTs with different number of galloyl moieties (GMs) on sucrase, maltase and α-amylase, and elucidate the relationship between α-glucosidase inhibitory activities and the number and connection mode of GMs. METHODS: Molecular docking and dynamics were used to study the binding mode and binding ability of five GTs against sucrase, maltase and α-amylase. Then, the inhibitory activities and inhibitory mechanisms of these compounds on sucrase, maltase and α-amylase in vitro were studied using inhibitory assay and enzyme inhibition kinetics. Further, the hypoglycemic effects in vivo of these compounds were demonstrated by three polysaccharides tolerance experiments on diabetes model mice. RESULTS: The results of molecular docking showed that these compounds could bind to enzymes through hydrogen bonds, hydrophobic interactions, etc. In addition, the α-glucosidase inhibition comparative studies in vitro and in vivo demonstrated that the inhibitory activities of these compounds on all three sucrase, maltase and α-amylase were ranked as TA ≈ PGG > TeGG > TGG > 1GG, and their inhibitory activities increases with the increase in the number of GMs. Moreover, the hypoglycemic effects of 1,2,3,4,6-pentagalloylglucose (PGG) and tannic acid (TA) in vitro and in vivo were also confirmed to be equivalent to or even stronger than that of acarbose. CONCLUSION: α-Glucosidase inhibitory activities in vitro and in vivo of GTs were positively correlated with the number of GTs, and the more the number, the stronger the activity. However, PGG with five GTs and TA with ten GTs showed almost identical α-glucosidase inhibitory activities, possibly due to the reduced binding force with the enzyme caused by spatial hindrance.