Replication Bypass of the N-(2-Deoxy-d-erythro-pentofuranosyl)-urea DNA Lesion by Human DNA Polymerase η.

Urea lesions in DNA arise from thymine glycol (Tg) or 8-oxo-dG; their genotoxicity is thought to arise in part due to their potential to accommodate the insertion of all four dNTPs during error-prone replication. Replication bypass with human DNA polymerase η (hPol η) confirmed that all four dNTPs were inserted opposite urea lesions but with purines exhibiting greater incorporation efficiency. X-ray crystal structures of ternary replication bypass complexes in the presence of Mg2+ ions with incoming dNTP analogs dAMPnPP, dCMPnPP, dGMPnPP, and dTMPnPP bound opposite urea lesions (hPol η·DNA·dNMPnPP complexes) revealed all were accommodated by hPol η. In each, the Watson-Crick face of the dNMPnPP was paired with the urea lesion, exploiting the ability of the amine and carbonyl groups of the urea to act as H-bond donors or acceptors, respectively. With incoming dAMPnPP or dGMPnPP, the distance between the imino nitrogen of urea and the N9 atoms of incoming dNMPnPP approximated the canonical distance of 9 Å in B-DNA. With incoming dCMPnPP or dTMPnPP, the corresponding distance of about 7 Å was less ideal. Improved base-stacking interactions were also observed with incoming purines vs pyrimidines. Nevertheless, in each instance, the α-phosphate of incoming dNMPnPPs was close to the 3'-hydroxyl group of the primer terminus, consistent with the catalysis of nucleotidyl transfer and the observation that all four nucleotides could be inserted opposite urea lesions. Preferential insertion of purines by hPol η may explain, in part, why the urea-directed spectrum of mutations arising from Tg vs 8-oxo-dG lesions differs.

Polymer Backbone Chemistry Shapes the Alkaline Stability of Metallopolymer Anion-Exchange Membranes.

Anion-exchange membrane fuel cells and water electrolyzers have garnered significant attention in past years due to their potential role in sustainable and affordable energy conversion and storage. However, the chemical stability of the polymeric anion-exchange membranes (AEMs), the key component in these devices, currently limits their lifespan. Recently, metallopolymers have been proposed as chemically stable alternatives to organic cations, using metal centers as ion transporters. In metallopolymer AEMs, various properties such as alkaline stability, water uptake, flexibility, and performance, are determined by both the metal complex and polymer backbone. Herein we present a systematic study investigating the influence of the polymer backbone chemistry on some of these properties, focusing on the alkaline stability of low-oxophilicity gold metallopolymers. Despite the use of a common N-heterocyclic carbene ligand, upon gold metalation using the same reaction conditions, different polymer backbones end up forming different gold complexes. These findings suggest that polymer chemistry affects the metalation reaction in addition to the other properties relevant to AEM performance.

Dietary inclusion of Clostridium butyricum cultures alleviated impacts of high-carbohydrate diets in largemouth bass (Micropterus salmoides).

A 60-d feeding trial was conducted to explore the potential regulatory effects of dietary Clostridium butyricum cultures (CBC) supplementation in high-carbohydrate diet (HCD) on carbohydrate utilisation, antioxidant capacity and intestinal microbiota of largemouth bass. Triplicate groups of largemouth bass (average weight 35·03 ± 0·04 g), with a destiny of twenty-eight individuals per tank, were fed low-carbohydrate diet and HCD supplemented with different concentration of CBC (0 %, 0·25 %, 0·50 % and 1·00 %). The results showed that dietary CBC inclusion alleviated the hepatic glycogen accumulation induced by HCD intake. Additionally, the expression of hepatic ampkα1 and insulin signaling pathway-related genes (ira, irb, irs, p13kr1 and akt1) increased linearly with dietary CBC inclusion, which might be associated with the activation of glycolysis-related genes (gk, pfkl and pk). Meanwhile, the expression of intestinal SCFA transport-related genes (ffar3 and mct1) was significantly increased with dietary CBC inclusion. In addition, the hepatic antioxidant capacity was improved with dietary CBC supplementation, as evidenced by linear decrease in malondialdehyde concentration and expression of keap1, and linear increase in antioxidant enzyme activities (total antioxidative capacity, total superoxide dismutase and catalase) and expression of antioxidant enzyme-related genes (nrf2, sod1, sod2 and cat). The analysis of bacterial 16S rRNA V3-4 region indicated that dietary CBC inclusion significantly reduced the enrichment of Firmicutes and potential pathogenic bacteria genus Mycoplasma but significantly elevated the relative abundance of Fusobacteria and Cetobacterium. In summary, dietary CBC inclusion improved carbohydrate utilization, antioxidant capacity and intestinal microbiota of largemouth bass fed HCD.

Dietary lipid sources affect growth performance, lipid deposition, antioxidant capacity and inflammatory response of largemouth bass (Micropterus salmoides).

The present study explored the effects of different lipid sources on growth performance, lipid deposition, antioxidant capacity, inflammatory response and disease resistance of largemouth bass (Micropterus salmoides). Four isonitrogenous (crude protein 50.46 %) and isolipidic (crude lipid 11.12 %) diets were formulated to contain 7 % of different oil sources including fish oil (FO) (control), soybean oil (SO), linseed oil (LO) and coconut oil (CO). Largemouth bass with initial body weight of 36.0 ± 0.2 g were randomly distributed into 12 tanks, with 30 fish per tank and 3 tanks per treatment. The fish were fed with the experiment diets twice daily for 8 weeks. The results indicated that the weight gain of largemouth bass fed the FO diet was significantly higher than that of fish fed the LO and CO diets. The liver crude lipid content in FO group was significantly higher than other groups, while the highest liver triglyceride content was showed in SO group and the lowest was detected in LO group. At transcriptional level, expression of lipogenesis related genes (pparγ, srebp1, fas, acc, dgat1 and dgat2) in the SO and CO group were significantly higher than the FO group. However, the expression of lipolysis and fatty acids oxidation related genes (pparα, cpt1, and aco) in vegetable oils groups were significantly higher than the FO group. As to the antioxidant capacity, vegetable oils significantly reduced the malondialdehyde content of largemouth bass. Total antioxidant capacity in the SO and LO groups were significantly increased compared with the FO group. Catalase in the LO group was significantly increased compared with the FO group. Furthermore, the ER stress related genes, such as grp78, atf6α, atf6ß, chop and xbp1 were significantly enhanced in the vegetable oil groups compared with the FO group. The activity of serum lysozyme in vegetable oil groups were significantly higher than in FO group. Additionally, the relative expression of non-specific immune related genes, including tlr2, mapk11, mapk13, mapk14, rela, tgf-ß1, tnfα, 5lox, il-1ß and il10, were all significantly increased in SO and CO groups compared to the other groups. In conclusion, based on the indexes including growth performance, lipid deposition, antioxidant capacity and inflammatory response, SO and LO could be alternative oil sources for largemouth bass.

Understanding the electrochemical properties of Mg-doped Li2MnO3: first-principles calculations.

Non-transition metal doping, especially for Mg, has been gradually employed to optimize the electrochemical performance of Li-rich cathode material Li2MnO3. However, the effects of Mg doping on the electrochemical behavior of Li2MnO3 have not been studied extensively. In this work, we investigate the effect of Mg doping at both the 2b (in the Li/Mn mixed layer) and 4h (in the Li layer) Li sites on the electrochemical properties of Li2MnO3 through first-principles calculations and ab initio molecular dynamics simulations. The local lattice structure, electronic density of states, Bader charge, delithiation voltage, lattice oxygen stability and Li diffusion kinetics are examined. Electronic structure analysis shows that Mg can activate the electrochemical activity of surrounding Mn by charge transfer, making Mn participate in charge compensation at the initial delithiation stage. Mg doping can also cause an increase in the average oxygen vacancy formation energy and hence depress the oxygen release during the delithiation process. Molecular dynamics simulations show that the diffusion kinetics of Li ions in Mg2b-Li2MnO3 is enhanced with respect to the undoped one, whereas Mg doped at the 4h site cannot improve the diffusion kinetics of Li ions. Further studies found that Mg doped at the 2b site results in a decrease in the energy barrier for the intra-layer diffusion and an increase in the energy barrier for the inter-layer diffusion of the nearby Li vacancies.

Temporal dynamic patterns of the ventromedial prefrontal cortex underlie the association between rumination and depression.

As a major contributor to the development of depression, rumination has proven linked with aberrant default-mode network (DMN) activity. However, it remains unclear how the spontaneous spatial and temporal activity of DMN underlie the association between rumination and depression. To illustrate this issue, behavioral measures and resting-state functional magnetic resonance images were connected in 2 independent samples (NSample1 = 100, NSample2 = 95). Fractional amplitude of low-frequency fluctuations (fALFF) and regional homogeneity (ReHo) were used to assess spatial characteristic patterns, while voxel-wise functional concordance (across time windows) (VC) and Hurst exponent (HE) were used to assess temporal dynamic patterns of brain activity. Results from both samples consistently show that temporal dynamics but not spatial patterns of DMN are associated with rumination. Specifically, rumination is positively correlated with HE and VC (but not fALFF and ReHo) values, reflecting more consistent and regular temporal dynamic patterns in DMN. Moreover, subregion analyses indicate that temporal dynamics of the ventromedial prefrontal cortex (VMPFC) reliably predict rumination scores. Furthermore, mediation analyses show that HE and VC of VMPFC mediate the association between rumination and depression. These findings shed light on neural mechanisms of individual differences in rumination and corresponding risk for depression.

A robust ultra-performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of 10 components in glutathione cycle.

Glutathione (GSH) is an important antioxidant that is generated and degraded via the GSH cycle. Quantification of the main components in the GSH cycle is necessary to evaluate the process of GSH. In this study, a robust ultra-performance liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of 10 components (GSH; γ-glutamylcysteine; cysteinyl-glycine; n-acetylcysteine; homocysteine; cysteine; cystine; methionine; glutamate; pyroglutamic acid) in GSH cycle was developed. The approach was optimized in terms of derivative, chromatographic, and spectrometric conditions as well as sample preparation. The unstable thiol groups of GSH, γ-glutamylcysteine, cysteinyl-glycine, n-acetylcysteine, cysteine, and homocysteine were derivatized by n-ethylmaleimide. The derivatized and underivatized analytes were separated on an amino column with gradient elution. The method was further validated in terms of selectivity (no interference), linearity (R2 > 0.99), precision (% relative standard deviation [RSD%] range from 0.57 to 10.33), accuracy (% relative error [RE%] range from -3.42 to 10.92), stability (RSD% < 5.68, RE% range from -2.54 to 4.40), recovery (RSD% range from 1.87 to 7.87) and matrix effect (RSD% < 5.42). The validated method was applied to compare the components in the GSH cycle between normal and oxidative stress cells, which would be helpful in clarifying the effect of oxidative stress on the GSH cycle.

The presence of a sinus tract is associated with reinfection after two-stage revision surgery for prosthetic hip joint infection: a case-control study.

BACKGROUND: Reinfection rates after two-stage revision (TSR) for prosthetic joint infection (PJI) range from 7.9 to 14%. Many factors, including sinus tracts, are associated with reinfection after this procedure. This study aimed to delineate whether the presence of sinus tract could increase reinfection rate after TSR and to investigate other potential risk factors for reinfection after TSR. METHODS: We conducted a case-control study by retrospectively reviewing patients who underwent TSR for prosthetic hip joint infection from 2002 to 2022. The case group included patients who developed reinfection after TSR, while the control group consisted of patients who did not experience reinfection. PJI and reinfection after TSR were defined based on Delphi-based international consensus criteria. Patient demographics, past medical history, clinical manifestations, laboratory results, interval between stages, microbiological culture results were collected. Univariate analyses were utilized to assess the effect of sinus tract on reinfection and to identify other risk factors for reinfection after TSR. RESULTS: Six patients with reinfection after TSR were included as the case group and 32 patients without reinfection were in the control group. Significant difference was observed in percentage of patients with sinus tracts between the two groups (67% in the case group versus 19% in the control group, p = 0.031, OR = 8.7). Significant difference was also found in percentage of patients with positive cultures of synovial fluid and synovium harvested during the first-stage revision between the two groups (100% in the case group versus 50% in the control group, p = 0.030). Additionally, patients in the case group had a significantly higher C-reactive protein (CRP) level prior to the second stage revision than that of patients in the control group (8.80 mg/L versus 2.36 mg/L, p = 0.005), despite normal CRP levels in all patients. CONCLUSIONS: Our study revealed that the presence of sinus tracts could significantly increase risk of postoperative reinfection after TSR. Positive cultures during the first stage revision and elevated CRP level prior to the second stage revision could also increase the risk of reinfection after TSR. Further studies with a larger sample size are required. TRIAL REGISTRATION: Retrospectively registered.

Integrating multi-index determination coupled with hierarchical cluster analysis to evaluate the quality consistency of PVE30, an anti-HSV "glycoprotein" macromolecule of Prunellae Spica.

INTRODUCTION: Prunellae Spica (PS), derived from the dried fruit spikes of Prunella vulgaris L., is a traditional Chinese medicinal herb. Our previous studies found that PVE30, a water-extracting ethanol-precipitating "glycoprotein" macromolecule of PS, was a potential anti-herpes simplex virus (HSV) candidate. However, due to the complex structure and diverse bioactivity of the "glycoprotein", ensuring its quality consistency across different batches of PVE30 becomes particularly challenging. This poses a significant hurdle for new drug development based on PVE30. OBJECTIVE: Our study aimed to integrate multi-index determination coupled with hierarchical cluster analysis (HCA) to holistically profile the quality consistency of "glycoprotein" in PVE30. METHODS: High-performance gel permeation chromatography with refractive index detector (HPGPC-RID) was used to characterise the molecular weight (Mw) distribution, HPLC-PDA was used to quantitatively analyse the composed monosaccharides and amino acids, and UV-VIS was used to quantify the contents of polysaccharides and proteins. Qualitative and quantitative consistency was analysed for each single index in 16 batches of PVE30, and a 16 × 38 data matrix, coupled with HCA, was used to evaluate the holistic quality consistency of PVE30. RESULTS: The newly developed and validated methods were exclusive, linear, precise, accurate, and stable enough to quantify multi-indexes in PVE30. Single-index analysis revealed that 16 batches of PVE30 were qualitatively consistent in Mw distribution, polysaccharides and proteins, and the composition of composed monosaccharides and amino acids but quantitatively inconsistent in the relative contents of some "glycoprotein" macromolecules, as well as the composed monosaccharides/amino acids. HCA showed that the holistic quality of PVE30 was inconsistent, the inconsistency was uncorrelated with the regions where PS was commercially collected, and the contents of 17 amino acids and 2 monosaccharides contributed most to the holistic quality inconsistency. CONCLUSION: Multi-index determination coupled with HCA was successful in evaluating the quality consistency of PVE30, and the significant difference in quantitative indices was not caused by the origin of PS. The cultivating basis should be confirmed for PVE30-based new drug development.

Integrating UPLC-QTOF-MS/MS and UPLC-DAD to evaluate the influence of sulfur-fumigated Paeoniae Radix Alba on the overall quality of three Si-Wu-Tang formulations.

INTRODUCTION: Sulfur-fumigation of Paeoniae Radix Alba (PRA) could induce the chemical transformation of its bioactive component paeoniflorin into a sulfur-containing derivative paeoniflorin sulfite, and thus alter the quality, bioactivities, pharmacokinetics, and toxicities of PRA. However, how sulfur-fumigated PRA (S-PRA) affects the quality of PRA-containing complex preparations has not been intensively evaluated. OBJECTIVES: We intend to evaluate the influence of S-PRA on the overall quality of three kinds of Si-Wu-Tang (SWT) formulations, i.e., decoction (SWT-D), granule (SWT-G), and mixture (SWT-M). MATERIAL AND METHODS: An UPLC-DAD multi-components quantification method was used to compare the transfer rates of paeoniflorin sulfite and other 10 bioactive components between S-PRA-containing and NS-PRA-containing SWT formulations. An UPLC-QTOF-MS/MS-based target metabolomics approach was applied to explore the differential sulfur-containing derivatives in S-PRA-containing SWT formulations. RESULTS: The transfer rates of paeoniflorin sulfite in three S-PRA-containing SWT formulations were all higher than 100%. Moreover, S-PRA also increased the transfer rate of 5-hydroxymethylfurfural, 1,2,3,4,6-O-pentagalloylglucose, whereas decreased that of paeoniflorin, albiflorin, and ferulic acid in three SWT formulations. Six pinane monoterpene glucoside sulfites originally identified in S-PRA, were also detectable in three S-PRA-containing SWT formulations. In addition, seven phenolic acid sulfites including (3Z)-6-sulfite-ligustilide, (3E)-6-sulfite-ligustilide, 6,8-disulfite-ligustilide, ferulic acid sulfite, neochlorogenic acid sulfite, chlorogenic acid sulfite, and angelicide sulfite (or isomer) were newly identified in these three S-PRA-containing formulations. CONCLUSION: S-PRA could differentially affect the transfer rate of paeoniflorin sulfite and other bioactive components during the preparation of three SWT formulations and subsequently the overall quality thereof.

Rationality of the ethanol precipitation process in modern preparation production of Zishui-Qinggan decoction evaluated by integrating UPLC-QTOF-MS/MS-based chemical profiling/serum pharmacochemistry and network pharmacology.

INTRODUCTION: Zishui-Qinggan decoction (ZQD) is a classical traditional Chinese medicine formula (TCMF) for alleviating menopausal symptoms (MPS) induced by endocrine therapy in breast cancer patients. In the production of TCMF modern preparations, ethanol precipitation (EP) is a commonly but not fully verified refining process. OBJECTIVES: Chemical profiling/serum pharmacochemistry and network pharmacology approaches were integrated for exploring the rationality of the EP process in the production of ZQD modern preparations. MATERIAL AND METHODS: Ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was applied to identify the chemical profiles and absorbed components of ZQD. Network pharmacology was used to identify targets and pathways related to MPS-relieving efficacy. RESULTS: The chemicals of ZQDs without/with EP process (referred to as ZQD-W and ZQD-W-P, respectively) were qualitatively similar with 89 and 87 components identified, respectively, but their relative contents were different; 51 components were detectable in the serum of rats orally administered with ZQD-W, whereas only 19 were detected in that administered with ZQD-W-P. Key targets, such as AKT1, and pathways, such as the PI3K-Akt signalling pathway, affected by ZQD-W and ZQD-W-P were similar, while the neuroactive ligand-receptor interaction pathway among others and the MAPK signalling pathway among others were specific pathways affected by ZQD-W and ZQD-W-P, respectively. The specifically absorbed components of ZQD-W could combine its specific key targets. CONCLUSION: The EP process quantitatively altered the chemical profiles of ZQD, subsequently affected the absorbed components of ZQD, and then affected the key targets and pathways of ZQD for relieving MPS. The EP process might result in variation of the MPS-relieving efficacy of ZQD, which deserves further in vivo verification.

Inositol Inclusion Affects Growth, Body Composition, Antioxidant Performance, and Lipid Metabolism of Largemouth Bass (Micropterus salmoides).

The present study explored the effects of inositol on growth performance, body composition, antioxidant performance, and lipid metabolism of largemouth bass (Micropterus salmoides). Six isonitrogenous and isolipidic diets containing 0 mg/kg (G1, control), 125 mg/kg (G2), 250 mg/kg (G3), 375 mg/kg (G4), 500 mg/kg (G5), and 625 mg/kg (G6) inositol were prepared and fed to cultured fish (initial weight: 110 ± 1 g) for 8 weeks in recirculating the aquaculture systems. The results indicated that compared with G1 group, the weight gain rate (WGR), specific growth rate (SGR), and feed efficiency rate (FER) in the G3 group were significantly higher. The crude lipid content of the whole fish and the liver of cultured fish was significantly reduced with increasing dietary inositol inclusion. However, no significant effects on moisture, crude protein, and ash contents of fish were observed among the different groups. Dietary inositol supplementation significantly increased muscular crude protein. However, muscular total lipid contents were decreased when the inclusion level was higher than 250 mg/kg (G3-G6 groups). As dietary inositol supplemental level increased, serum triglyceride (TG), and cholesterol (TC) contents showed an increasing trend and reached the maximum value in the G3 group. Additionally, serum low-density lipoprotein cholesterol (LDL-C) in G2, G3, G4, and G5 groups was significantly upregulated by increasing inositol. While, there was no significant change in serum high-density lipoprotein cholesterol (HDL-C) among the treatments. Inositol inclusion also significantly reduced the serum alkaline phosphatase (AKP), glutamic-pyruvic transaminase (ALT), and glutamic-oxaloacetic transaminase (AST) activities as well as serum malondialdehyde (MDA) content but significantly increased serum catalase (CAT), superoxide dismutase (SOD) activities, and total antioxidant capacity (T-AOC). Compared with the control group, the activities of hepatic total lipase (TL) and lipoprotein lipase (LPL) were significantly elevated in the G3, G4, and G5 groups. Above all, dietary inositol supplementation could improve growth performance and antioxidant capacity, and reduce the liver fat content of largemouth bass, and the optimal supplementation level of inositol in feed is estimated to be 250.31-267.27 mg/kg.

PI3KR1 and AKT1 in largemouth bass (Micropterus salmoides): molecular cloning, characterization, and its involvement in the alleviation of hepatic glycogen deposition caused by insulin inclusion in vitro.

In this study, the full-length cDNA sequences of the phosphatidylinositol-3-kinase p85 alpha (PI3KR1) and serine/threonine kinase 1 (AKT1) genes in largemouth bass (Micropterus salmoides) were obtained using the rapid amplification of cDNA ends (RACE) method. Sequence analysis revealed that the cloned sequences of PI3KR1 and AKT1 are 4170 bp and 3672 bp in length, with open reading frames (ORFs) of 1389 bp and 1422 bp encoding 462 and 473 amino acids, respectively. Sequence alignment and evolutionary tree analysis indicated their close relationship to other teleosts, especially those with similar feeding habits. Tissue distribution demonstrated widespread distribution of both genes in various tissues, with the highest abundance in the liver. Further results found that the upregulation of the expression of p-PI3KR1, p-AKT1, p-FoxO1, and GLUT2 proteins by insulin, while suppressing the expression of the total FoxO1 protein, effectively triggers a significant activation of the PI3KR1-AKT1 insulin signaling pathway. Meanwhile, the mRNA levels of the key glycolytic genes, including glucokinase (gk), pyruvate kinase (pk), and phosphofructokinase liver type (pfkl), have been enhanced evidently. In contrast, the expression of gluconeogenic genes such as phosphoenolpyruvate carboxykinase (pepck), glucose-6-phosphatase catalytic subunit (g6pc), and fructose-1,6-bisphosphatase-1 (fbp1) has been notably down-regulated. In addition, insulin treatment promoted the phosphorylation of glycogen phosphorylase (PYGL) and the dephosphorylation of glycogen synthase (GS), and the glycogen content in the insulin-treated group was remarkably reduced compared to the control group. Overall, our study indicates that the activation of PI3KR1-AKT1 insulin signaling pathway represses the hepatic glycogen deposition via the regulation of glycolysis and gluconeogenesis, which provides some new insights into nutritional strategy to effectively regulate the glucose metabolism in carnivorous fish.

The preferential utilization of hepatic glycogen as energy substrates in largemouth bass (Micropterus salmoides) under short-term starvation.

To elucidate the underlying mechanism of the energy metabolism in largemouth bass (Micropterus salmoides), cultured fish (initial body weight: 77.57 ± 0.75 g) in the present study were starved for 0 h, 12 h, 24 h, 48 h, 96 h and 192 h, respectively. The proximate composition analysis showed that short-term starvation induced a significant up-regulation in crude protein proportion in hepatic of cultured fish (P < 0.05). However, short-term starvation significantly decreased the hepatosomatic index and the viscerosomatic index of cultured fish (P < 0.05). The exact hepatic glycogen content in the group starved for 92 h presented remarkable decrease (P < 0.05). Meanwhile, compared with the weight change of lipid and protein (mg) in hepatic (y = 0.0007x2 - 0.2827x + 49.402; y = 0.0013x2 - 0.5666x + 165.31), the decreasing trend of weight in glycogen (mg) was more pronounced (y = 0.0032x2 - 1.817x + 326.52), which suggested the preferential utilization of hepatic glycogen as energy substrates under short-term starvation. Gene expression analysis revealed that the starvation down-regulated the expression of insulin-like growth factor 1 and genes of TOR pathway, such as target of rapamycin (tor) and ribosomal protein S6 (s6) (P < 0.05). In addition, the starvation significantly enhanced expression of lipolysis-related genes, including hormone-sensitive lipase (hsl) and carnitine palmitoyl transferase I (cpt1), but down-regulated lipogenesis as indicated by the inhibited expression of fatty acids synthase (fas), acetyl-CoA carboxylase 1 (acc1) and acetyl-CoA carboxylase 2 (acc2) (P < 0.05). Starvation of 24 h up-regulated the expression of glycolysis genes, glucokinase (gk), phosphofructokinase liver type (pfkl) and pyruvate kinase (pk), and then their expression returned to the normal level. Meanwhile, the expression of gluconeogenesis genes, such as glucose-6-phosphatase catalytic subunit (g6pc), fructose-1,6-bisphosphatase-1 (fbp1) and phosphoenolpyruvate carboxy kinase (pepck), was significantly inhibited with the short-term starvation (P < 0.05). In conclusion, short-term starvation induced an overall decline in growth performance, but it could deplete the hepatic glycogen accumulation and mobilize glycogen for energy effectively.

The gut microbiome: an important role in neurodegenerative diseases and their therapeutic advances.

There are complex interactions between the gut and the brain. With increasing research on the relationship between gut microbiota and brain function, accumulated clinical and preclinical evidence suggests that gut microbiota is intimately involved in the pathogenesis of neurodegenerative diseases (NDs). Increasingly studies are beginning to focus on the association between gut microbiota and central nervous system (CNS) degenerative pathologies to find potential therapies for these refractory diseases. In this review, we summarize the changes in the gut microbiota in Alzheimer's disease, Parkinson's disease, multiple sclerosis, and amyotrophic lateral sclerosis and contribute to our understanding of the function of the gut microbiota in NDs and its possible involvement in the pathogenesis. We subsequently discuss therapeutic approaches targeting gut microbial abnormalities in these diseases, including antibiotics, diet, probiotics, and fecal microbiota transplantation (FMT). Furthermore, we summarize some completed and ongoing clinical trials of interventions with gut microbes for NDs, which may provide new ideas for studying NDs.

Iron and nitrogen co-modified multi-walled carbon nanotubes for efficient electrocatalytic oxygen reduction.

Multi-walled carbon nanotubes (MWCNTs) with one-dimensional nanostructure are an ideal support for oxygen reduction reaction (ORR) catalysts thanks to their intrinsic outstanding electrical conductivity and high specific surface area. Iron and nitrogen doping could alter the local electronic structure and therefore enhance the ORR activity of MWCNTs, but the preparation process always includes complicated growth conditions and post-treatment. Herein, an iron and nitrogen co-modified multi-walled carbon nanotubes (Fe-N-MWCNTs) with hierarchical nanostructure is engineered and synthesized via a simple two-step pyrolysis approach. Large specific surface area, low resistivity, and intensified charge density near the Fermi level synergistically endow the Fe-N-MWCNTs with outstanding ORR activity. The optimal Fe-N-MWCNTs exhibit a higher onset potential value of 0.92 V (versus RHE) and half-wave potential (E1/2) of 0.85 V (versus RHE) in 0.1 M KOH medium, which exceeds the benchmark Pt/C electrocatalyst (E1/2= 0.84 V). This strategy of modifying MWCNTs support by a simple calcination process provides a feasible method to prepare cost-efficient ORR electrocatalysts.

Artificially Intelligent Three-Dimensionally-Printed Patient-Specific Instrument Improves Total Hip Arthroplasty Accuracy.

BACKGROUND: Patient-specific instrumentation (PSI) has the potential to improve the accuracy of implant positioning in total hip arthroplasty (THA). This prospective clinical study aimed to develop artificial intelligence to increase PSI production efficiency and assess accuracy, clinical outcomes, and learning curves. METHODS: A convolutional neural network was applied to automatically process computer tomography images. PSI size and position were designed to guide the acetabular preparation and femoral neck resection. Thirty patients who underwent PSI-assisted THAs were matched to thirty patients who underwent free-hand THAs, and the component positions, as well as radiographic and clinical outcomes were analyzed. RESULTS: PSI-assisted THA was significantly more accurate than free-hand THA at achieving the target component position. The mean absolute errors of cup inclination (P = .004) and anteversion (P < .001) were significantly smaller in the PSI group with fewer outliers. Calcar length (P = .002) and neck length (P = .026) were also more accurate in the PSI group. The leg length discrepancy was significantly lower in the PSI group (P = .002). There were no significant differences in operation time, blood loss, leg length discrepancy, or cup position among the first, second, and last 10 cases. CONCLUSION: PSI-assisted THA offered more accurate component positions and better radiographic outcomes than free-hand THA. There was no evidence of a learning curve. Our findings suggest that PSI is a convenient and practical option to help surgeons achieve accurate surgical outcomes.

Acetabular Diameter Assessment and Three-Dimensional Simulation for Acetabular Reconstruction in Dysplastic Hips.

BACKGROUND: This study aimed to investigate the relationship between acetabular width, three-dimensional (3D) simulation, and surgical results in total hip arthroplasty patients who have developmental dysplasia of the hip (DDH). METHODS: This retrospective study included 216 DDH cases. Inner and outer acetabular width (OAW) was measured at the plane passing through the center of acetabular fossa. 3D simulation and 2D standard templating were performed. The actual cup size and the use of augments during surgery were recorded. Association among the indices and their distribution in different types of DDH were analyzed. RESULTS: A difference of 13 to 14 millimeters (mm) was found between the inner acetabular width and actual cup size used in type II, III, and IV cases, while the difference was 0.2 to 3.6 mm for OAW. The accuracy of 2D templating and 3D simulation in predicting cup size was comparable in Crowe type I (86.5 versus 76%, P = .075), type II (72.7 versus 51.5%, P = .127), and type III (93.3 versus 66.7%, P = .169). The 3D simulation was significantly more accurate in Crowe type IV (89.1% versus 60.9%, P = .001). Augments and bone grafts were significantly more commonly used in type II (25%) than in the other types (0 to 6.5%). CONCLUSION: OAW more accurately predicted actual cup size than inner acetabular width. The supero-lateral acetabular bone defects in type II cases require additional attention. Compared with 2D templating, 3D simulation is more accurate in predicting actual cup size in dysplastic hips with severe deformity and may be recommended in these selected cases, especially for Crowe IV patients.

Quality consistency evaluation of commercial Prunellae Spica by integrating determination of secondary metabolites and saccharides.

INTRODUCTION: Prunellae Spica (PS) is a commonly used medicinal herb in China. Secondary metabolites and saccharides are major bioactive components of PS. However, holistic quality consistency of commercial PS is ambiguous due to lack of comprehensive evaluation methods and reliable quality control markers. OBJECTIVES: Integrating multiple chromatographic and chemometric methods to comprehensively evaluate the holistic quality of PS. MATERIAL AND METHODS: Ultrahigh-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-TQ-MS/MS) was applied to quantify 12 secondary metabolites of PS. High-performance liquid chromatography coupled with photodiode array/evaporative light scattering detection (HPLC-PDA/ELSD) and high-performance gel permeation chromatography (HPGPC) methods were used to characterise the saccharides. Multivariate statistical analysis was adopted to evaluate the quality consistency of commercial PS and explore the potential quality control markers. RESULTS: The contents of secondary metabolites and saccharides were significantly different among commercial PS. All samples could be classified into three groups with ferulic acid, protocatechualdehyde, gallic acid, ursolic acid/oleanolic acid, sucrose, p-coumaric acid, chlorogenic acid as the major contributing components responsible for the difference. The content of rosmarinic acid was correlated with that of betulinic acid, hyperposide, chlorogenic acid, rutin, caffeic acid, p-coumaric acid and glucose, whereas polysaccharides, ferulic acid, protocatechualdehyde and ursolic acid/oleanolic acid, quercetin, sucrose and majority monosaccharides were not. CONCLUSION: The holistic quality of commercial PS was inconsistent. Together with rosmarinic acid, ferulic acid, protocatechualdehyde, ursolic acid/oleanolic acid, polysaccharides and sucrose might be recommended as potential quality control markers for the holistic quality control of PS.

Integrating serum pharmacochemistry and network pharmacology to identify chemical markers for quality control of Apocyni Veneti Folium.

INTRODUCTION: Apocyni Veneti Folium (AVF) is a commonly used traditional Chinese medicinal herb for the treatment of hypertension. Chemical markers are crucial for the quality control of herbal medicines; however, the therapeutic components of AVF remain to be well elucidated. OBJECTIVES: This study was intended to integrate serum pharmacochemistry and network pharmacology to identify chemical markers of AVF and establish an efficacy-related quality control method of AVF. MATERIAL AND METHODS: Ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was applied to identify the absorbed AVF constituents in rat serum. Network pharmacology was further used to identify anti-hypertension-related chemical markers. Subsequently, a quantitative method was established using UPLC with diode array detection (DAD) and applied for quality evaluation of commercial AVF samples. RESULTS: Thirteen prototype constituents were unequivocally or tentatively characterized in serum samples, among which quercetin, kaempferol, hyperoside, isoquercitrin, chlorogenic acid, cryptochlorogenic acid, and neochlorogenic acid were identified as dominant chemicals related to anti-hypertensive efficacy. The quantitative data showed that the total contents of seven marker components even showed 2-fold variation among 14 batches of commercial AVF samples with RSD values ranging from 12.15% to 75.61%. Hierarchical cluster analysis and heatmap analysis showed that 14 batches of commercial AVF samples could be divided into three main groups. CONCLUSION: The chemical markers obtained from this study could be applicable for efficacy-related quality control of AVF.