Structure of the Angiotensin receptor revealed by serial femtosecond crystallography.

Angiotensin II type 1 receptor (AT(1)R) is a G protein-coupled receptor that serves as a primary regulator for blood pressure maintenance. Although several anti-hypertensive drugs have been developed as AT(1)R blockers (ARBs), the structural basis for AT(1)R ligand-binding and regulation has remained elusive, mostly due to the difficulties of growing high-quality crystals for structure determination using synchrotron radiation. By applying the recently developed method of serial femtosecond crystallography at an X-ray free-electron laser, we successfully determined the room-temperature crystal structure of the human AT(1)R in complex with its selective antagonist ZD7155 at 2.9-Å resolution. The AT(1)R-ZD7155 complex structure revealed key structural features of AT(1)R and critical interactions for ZD7155 binding. Docking simulations of the clinically used ARBs into the AT(1)R structure further elucidated both the common and distinct binding modes for these anti-hypertensive drugs. Our results thereby provide fundamental insights into AT(1)R structure-function relationship and structure-based drug design.

Structural insights into angiotensin receptor signaling modulation by balanced and biased agonists.

The peptide hormone angiotensin II regulates blood pressure mainly through the type 1 angiotensin II receptor AT1 R and its downstream signaling proteins Gq and ß-arrestin. AT1 R blockers, clinically used as antihypertensive drugs, inhibit both signaling pathways, whereas AT1 R ß-arrestin-biased agonists have shown great potential for the treatment of acute heart failure. Here, we present a cryo-electron microscopy (cryo-EM) structure of the human AT1 R in complex with a balanced agonist, Sar1 -AngII, and Gq protein at 2.9 Å resolution. This structure, together with extensive functional assays and computational modeling, reveals the molecular mechanisms for AT1 R signaling modulation and suggests that a major hydrogen bond network (MHN) inside the receptor serves as a key regulator of AT1 R signal transduction from the ligand-binding pocket to both Gq and ß-arrestin pathways. Specifically, we found that the MHN mutations N1113.35 A and N2947.45 A induce biased signaling to Gq and ß-arrestin, respectively. These insights should facilitate AT1 R structure-based drug discovery for the treatment of cardiovascular diseases.

Ccq1 restrains Mre11-mediated degradation to distinguish short telomeres from double-strand breaks.

Telomeres protect chromosome ends and are distinguished from DNA double-strand breaks (DSBs) by means of a specialized chromatin composed of DNA repeats bound by a multiprotein complex called shelterin. We investigated the role of telomere-associated proteins in establishing end-protection by studying viable mutants lacking these proteins. Mutants were studied using a Schizosaccharomyces pombe model system that induces cutting of a 'proto-telomere' bearing telomere repeats to rapidly form a new stable chromosomal end, in contrast to the rapid degradation of a control DSB. Cells lacking the telomere-associated proteins Taz1, Rap1, Poz1 or Rif1 formed a chromosome end that was stable. Surprisingly, cells lacking Ccq1, or impaired for recruiting Ccq1 to the telomere, converted the cleaved proto-telomere to a rapidly degraded DSB. Ccq1 recruits telomerase, establishes heterochromatin and affects DNA damage checkpoint activation; however, these functions were separable from protection of the new telomere by Ccq1. In cells lacking Ccq1, telomere degradation was greatly reduced by eliminating the nuclease activity of Mre11 (part of the Mre11-Rad50-Nbs1/Xrs2 DSB processing complex), and higher amounts of nuclease-deficient Mre11 associated with the new telomere. These results demonstrate a novel function for S. pombe Ccq1 to effect end-protection by restraining Mre11-dependent degradation of the DNA end.

Structures of human gastrin-releasing peptide receptors bound to antagonist and agonist for cancer and itch therapy.

Gastrin releasing peptide receptor (GRPR), a member of the bombesin (BBN) G protein-coupled receptors, is aberrantly overexpressed in several malignant tumors, including those of the breast, prostate, pancreas, lung, and central nervous system. Additionally, it also mediates non-histaminergic itch and pathological itch conditions in mice. Thus, GRPR could be an attractive target for cancer and itch therapy. Here, we report the inactive state crystal structure of human GRPR in complex with the non-peptide antagonist PD176252, as well as two active state cryo-electron microscopy (cryo-EM) structures of GRPR bound to the endogenous peptide agonist gastrin-releasing peptide and the synthetic BBN analog [D-Phe6, ß-Ala11, Phe13, Nle14] Bn (6-14), in complex with Gq heterotrimers. These structures revealed the molecular mechanisms for the ligand binding, receptor activation, and Gq proteins signaling of GRPR, which are expected to accelerate the structure-based design of GRPR antagonists and agonists for the treatments of cancer and pruritus.

SUPPRESSOR OF FRIGIDA 4 cooperates with the histone methylation reader EBS to positively regulate root development.

Maintenance and homeostasis of the quiescent center (QC) in Arabidopsis (Arabidopsis thaliana) root apical meristems are critical for stem cell organization and root development. Despite great progress in relevant research, the molecular mechanisms that determine the root stem cell fate and QC still need further exploration. In Arabidopsis, SUPPRESSOR OF FRIGIDA 4 (SUF4) encodes a C2H2-type zinc finger protein that represses flowering by transcriptional activation of FLOWERING LOCUS C (FLC) through the FRIGIDA (FRI) pathway, and EARLY BOLTING IN SHORT DAYS (EBS) is a bivalent histone reader that prevents premature flowering. Here, we found that SUF4 directly interacts with EBS in vivo and in vitro. Loss of function of SUF4 and/or EBS resulted in disorganization of the QC, aberrant cell division, and stunted root growth. RNA-seq and reverse transcription quantitative real-time polymerase chain reaction analysis revealed that SUF4 and EBS coregulate many root development-related genes. A series of biochemical analyses demonstrated that SUF4 directly binds to the promoter of SCARECROW (SCR), which encodes a key regulator of root development. Chromatin immunoprecipitation assay indicated that both SUF4 and EBS are recruited to the SCR locus in an interdependent manner to promote H3K4me3 levels and suppress H3K27me3 levels, thereby activating the expression of SCR. These findings improve our understanding of the function of SUF4 and EBS and provide insights into the molecular mechanism that couples a transcription factor and a histone methylation reader to modulate QC specification and root development in Arabidopsis.

Drp1 acetylation mediated by CDK5-AMPK-GCN5L1 axis promotes cerebral ischemic injury via facilitating mitochondrial fission.

The aberrant acetylation of mitochondrial proteins is involved in the pathogenesis of multiple diseases including neurodegenerative diseases and cerebral ischemic injury. Previous studies have shown that depletion of mitochondrial NAD+, which is necessary for mitochondrial deacetylase activity, leads to decreased activity of mitochondrial deacetylase and thus causes hyperacetylation of mitochondrial proteins in ischemic brain tissues, which results in altered mitochondrial dynamics. However, it remains largely unknown about how mitochondrial dynamics-related protein Drp1 is acetylated in ischemic neuronal cells and brain tissues. Here, we showed that Drp1 and GCN5L1 expression was up-regulated in OGD-treated neuronal cells and ischemic brain tissues induced by dMCAO, accompanied by the increased mitochondrial fission, mtROS accumulation, and cell apoptosis. Further, we confirmed that ischemia/hypoxia promoted Drp1 interaction with GCN5L1 in neuronal cells and brain tissues. GCN5L1 knockdown attenuated, while its overexpression enhanced Drp1 acetylation and mitochondrial fission, indicating that GCN5L1 plays a crucial role in ischemia/hypoxia-induced mitochondrial fission by acetylating Drp1. Mechanistically, ischemia/hypoxia induced Drp1 phosphorylation by CDK5 upregulation-mediated activation of AMPK in neuronal cells, which in turn facilitated the interaction of GCN5L1 with Drp1, thus enhancing Drp1 acetylation and mitochondrial fission. Accordingly, inhibition of AMPK alleviated ischemia/hypoxia- induced Drp1 acetylation and mitochondrial fission and protected brain tissues from ischemic damage. These findings provide a novel insight into the functional roles of GCN5L1 in regulating Drp1 acetylation and identify a previously unrecognized CDK5-AMPK-GCN5L1 pathway that mediates the acetylation of Drp1 in ischemic brain tissues.

Self-Oxidated Hybrid Conductive Network Enables Efficient Electrochemical Lithium Extraction Under High-Altitude Environment.

The electrochemical deintercalation method has been considered as an effective way to address the demand for lithium resources due to its environmental friendliness, high selectivity, and high efficiency. However, the performance of electrochemical lithium extraction is closely dependent on the electrode material and needs to be compatible under plateau environments with high-altitude and low-temperature. Herein, an in situ self-oxidation method is conducted to construct a hybrid conductive network on the surface of LiFePO4 (LFP-HN). The introduction of a hybrid conductive network enhanced the interfacial electron/lithium-ion transfer. In addition, structural stability is strengthened through suppressing the intercalation of impurity cations. Consequently, the LFP-HN delivered extremely high lithium extraction capacity (27.42 mg g-1), low energy consumption (4.91 Wh mol-1), and superior purity (91.05%) in Baqiancuo real brine (4788 m, -10 °C). What's more, LFP-HN-based large-scale prototypes are constructed and operated at Baqiancuo, which is calculated to extract 25 kg Lithium Carbonate Equivalent per cycle (4.55 h, 100 pairs of plates). Based on the excellent performance, the modification strategy developed in this work can be a promising solution for industrial lithium extraction under high-altitude environment.

In Situ Construction of Bimetallic Selenides Heterogeneous Interface on Oxidation-Stable Ti3C2Tx MXene Toward Lithium Storage with Ultrafast Charge Transfer Kinetics.

Ti3C2Tx (MXene) is widely acknowledged as an excellent substrate for constructing heterogeneous structures with transition metal chalcogenides (TMCs) for boosting the electrochemical performance of lithium-ion storage. However, conventional synthesis strategies inevitably lead to poor electrochemical charge transfer due to Ti3C2Tx-derived TiO2 at the heterogeneous interface between Ti3C2Tx and TMCs. Here, an innovative in situ selenization strategy is proposed to replace the originally generated TiO2 on Ti3C2Tx with metallic TiSe2 interphase, clearing the bottleneck of slow charge transfer barrier caused by MXene oxidation. The construction of bimetallic selenide formed by CoSe2 and TiSe2 generates intrinsic electric fields to guide the fast ion diffusion kinetics in a heterogeneous interface. Additionally, the CoSe2/TiSe2/Ti3C2Tx heterogeneous structure with enhanced structural stability and improved rate performance is confirmed by both experiments and theoretical calculations. The engineered heterogeneous structure exhibits an ultra-high pseudocapacitance contribution (73.1% at 0.1 mV s-1), rendering it well-suited to offset the kinetics differences between double-layer materials. The assembled lithium-ion capacitor based on CoSe2/TiSe2/Ti3C2Tx possesses a high energy density and an ultralong life span (89.5% after 10 000 times at 2 A g-1). This devised strategy provides a feasible solution for utilizing the performance advantages of MXene substrates in lithium storage with ultrafast charge transfer kinetics.

Prognostic significance of tubulointerstitial macrophage density in MPO-ANCA-associated glomerulonephritis: implications for renal outcomes.

OBJECTIVE: This study aimed to evaluate the density of tubulointerstitial macrophages with renal outcomes in patients with myeloperoxidase (MPO)-ANCA associated glomerulonephritis (MPO-ANCA GN). METHODS: This study analyzed patients with MPO-ANCA GN who had renal biopsies at Jinling Hospital. It looked at the density of CD68+ macrophages in the tubulointerstitium and examined correlations with serum creatinine levels, urinary protein levels, treatment regimen, and renal histologic class. The study used KM curves to show the impact of these factors on renal prognosis and conducted multivariate analyses with Cox proportional hazards regression models. RESULTS: A total of 172 patients with MPO-ANCA GN (median age: 50 y, 43.6% male) were included. Stratification of the cohort into tertiles was based on tubulointerstitial macrophage density. Significant differences in serum creatinine levels, induction treatment regimen, the rates of ESKD, and renal histologic class were observed between the three groups. Correlation analysis showed that induction treatment regimen and renal histologic class were correlated with tubulointerstitial macrophage density. Kaplan-Meier curves illustrated patients with a lower presence of CD68+ macrophages in the tubulointerstitium experienced significantly better renal survival compared with those with a higher presence. The higher levels of CD68+ macrophage infiltration were significantly associated with adverse renal outcomes. This association persisted after adjusting for potential confounders including baseline serum creatinine, histopathological class, and induction therapy modalities. CONCLUSIONS: The results of our study provide insight into the prognostic significance of macrophage infiltration in the tubulointerstitium in MPO-ANCA GN.

Comparison of the Value of Four Objective Nutritional Indices in Assessing the Long-Term Prognosis of Elderly Patients with Heart Failure with Preserved Ejection Fraction.

Background: The long-term prognosis of heart failure with preserved ejection fraction (HFpEF) is influenced by malnutrition. Currently, there's a deficit in objective and comprehensive nutritional assessment methods to evaluate the nutritional status and predicting the long-term outcomes of HFpEF patients. Methods: Our retrospective study included two hundred and eighteen elderly HFpEF patients admitted to the cardiovascular ward at the Air Force Medical Centre from January 2016 to December 2021. Based on follow-up outcomes, patients were categorized into all-cause death (99 cases) and Survival (119 cases) groups. We compared general data, laboratory results, and nutritional indexes between groups. Differences in subgroups based on Triglyceride-Total Cholesterol-Body Weight Index (TCBI), Geriatric Nutritional Risk Index (GNRI), Prognostic Nutritional Index (PNI), and Controlled Nutrition Score (CONUT) were analyzed using Kaplan-Meier survival curves and log-rank test. COX regression was used to identify all-cause mortality risk factors, and the predictive accuracy of the four nutritional indices was assessed using receiver operating characteristic (ROC) curves and Delong test analysis. Results: A total of 101 (45.41%) HFpEF patients experienced all-cause mortality during 59.02 ± 1.79 months of follow-up. The all-cause mortality group exhibited lower GNRI and PNI levels, and higher CONUT levels than the Survival group (p < 0.05). Kaplan-Meier analysis revealed lower cumulative survival in the low GNRI ( ≤ 96.50) and low PNI ( ≤ 43.75) groups, but higher in the low CONUT ( ≤ 2) group, compared to their respective medium and high-value groups. Multifactorial COX regression identified low PNI ( ≤ 43.75) as an independent all-cause mortality risk factor in elderly HFpEF patients. ROC and Delong's test indicated PNI (area under the curve [AUC] = 0.698, 95% confidence interval [CI] 0.629-0.768) as a more effective predictor of all-cause mortality than TCBI (AUC = 0.533, 95% CI 0.456-0.610) and CONUT (AUC = 0.621, 95% CI 0.547-0.695; p < 0.05). However, there was no significant difference compared to GNRI (AUC = 0.663, 95% CI 0.590-0.735; p > 0.05). Conclusions: In elderly HFpEF patients a PNI ≤ 43.75 was identified as an independent risk factor for all-cause mortality. Moreover, PNI demonstrates superior prognostic performance in predicting all-cause mortality in elderly patients with HFpEF when compared to TCBI, GNRI, and COUNT.

Large-Scale Production and Integrated Application of Micro-Supercapacitors.

Micro-supercapacitors, emerging as promising micro-energy storage devices, have attracted significant attention due to their unique features. This comprehensive review focuses on two key aspects: the scalable fabrication of MSCs and their diverse applications. The review begins by elucidating the energy storage mechanisms and guiding principles for designing high-performance devices. It subsequently explores recent advancements in scalable fabrication techniques for electrode materials and micro-nano fabrication technologies for micro-devices. The discussion encompasses critical application domains, including multifunctional MSCs, energy storage integration, integrated power generation, and integrated applications. Despite notable progress, there are still some challenges such as large-scale production of electrode material, well-controlled fabrication technology, and scalable integrated manufacture. The summary concludes by emphasizing the need for future research to enhance micro-supercapacitor performance, reduce production costs, achieve large-scale production, and explore synergies with other energy storage technologies. This collective effort aims to propel MSCs from laboratory innovation to market viability, providing robust energy storage solutions for MEMS and portable electronics.

A novel dual-target Septin9 methylation assay for improved detection of early-stage colorectal cancer and high-grade intraepithelial neoplasia.

BACKGROUND: Colorectal cancer (CRC) ranks as the third most common malignancies in the world, and periodic examination of the patient is advantageous in reducing the mortality of CRC. The first blood-based Septin9 gene methylation assay which recognized by the US FDA for CRC examination was Epi proColon. However, this assay was not broadly applied in the current clinical guideline because of its relatively lower sensitivity in the detection of early-stage CRC. METHODS: This study aimed at developing a new multiplex Septin9 methylation assay (ColonUSK) which simultaneously evaluates two CpG-rich subregions in the promoter of the Septin9 gene and an internal control in a single reaction. ColonUSK proved increased sensitivity, with a detection limit as low as 12pg of the positive DNA compared with the Septin9 assay targeting one CpG-rich subregion. 1366 subjects were prospectively recruited from four comprehensive hospitals in China in an opportunistic screening study for assessing its value in CRC detection. Blind testing was developed to evaluate ColonUSK in comparison with clinical examination using clinical gold standard such as colonoscopy. RESULTS: The assay demonstrates clinical sensitivity for diagnosing colorectal cancer (CRC) and advanced adenoma at rates of 77.34% and 25.26%, respectively. Furthermore, ColonUSK exhibits a high degree of specificity for non-CRC cases (95.95%) clinically. Significantly, the detection rate of cases in high-grade intraepithelial neoplasia increased to 54.29%. The value for the assay in the Kappa test was 0.76, showing a high degree of consistency between ColonUSK and clinical gold standard. CONCLUSIONS: ColonUSK indicated moderate diagnostic value and could become a non-invasive detection way for CRC. The implementation of the ColonUSK assay has the capacity to markedly enhance CRC screening practices.

Transcatheter aortic valve implantation for severe aortic regurgitation using the J-Valve system: A midterm follow-up study.

BACKGROUND: Transcatheter aortic valve implantation (TAVI) is a well-established intervention for severe aortic valve stenosis. However, its application for severe aortic regurgitation (AR) is still under evaluation. This study aims to present the 3-year follow-up outcomes of the J-Valve system in managing severe AR. AIMS: The aim of this study was to evaluate the mid-term efficacy and durability of the J-Valve system in the treatment of severe AR and to provide new information on this intervention. METHODS: In this retrospective, single-center study, we evaluated the prognostic outcomes of patients with AR, who underwent treatment with the J-Valve system at Nanjing Drum Tower Hospital. Consecutive patients who were treated with the J-Valve were included in the analysis. The study focused on the echocardiographic follow-up to assess the effectiveness and durability of the J-Valve system in managing AR. RESULTS: From January 2018 to December 2022, 36 high-risk AR patients treated with the J-Valve system had a procedural success rate of 97.2%, with one case requiring open-heart surgery due to valve displacement. Significant improvements were observed in left ventricular diameter (from 63.50 [58.75-69.50] mm to 56.50 [53.00-60.50] mm, p < 0.001) and left atrial diameter (from 44.00 [40.00-45.25] mm to 39.00 [36.75-41.00] mm, p = 0.003) postsurgery. All patients completed the 1-year follow-up, with an overall mortality rate of 2 out of 36 (5.6%). Among the surviving patients, there was one case of III° atrioventricular block and one case of stroke, both occurring within 90 days postsurgery. After a 3-year follow-up, 15.0% of patients had mild or moderate valvular regurgitation, with no cases of moderate or severe paravalvular leak. Additionally, 89.5% of patients were classified as New York Heart Association class I or II, showing significantly enhanced cardiac function. CONCLUSION: The J-Valve system has shown positive therapeutic outcomes in treating AR, with notable effectiveness in managing the condition and significant improvements in heart failure symptoms and cardiac remodeling. However, due to the limited sample size and partial follow-up data, it is important to emphasize the need for further research with comprehensive long-term follow-up, to fully validate these results.

Non-small cell lung cancer patient with a rare UGP2-ALK fusion protein responded well to alectinib: a case report.

Several rare anaplastic lymphoma kinase (ALK) fusions have been identified in patients with non-small cell lung cancer (NSCLC); however, their treatment is not currently uniform. alectinib has been commonly used to treat rare ALK fusions in patients with NSCLC. This is the first study to report the occurrence of a uridine diphosphate-glucose pyrophosphorylase 2 (UGP2)-ALK fusion in a patient with NSCLC. The patient, who was hospitalized because of shortness of breath lasting 20 days, showed hydrothorax of the left lung under a computerized tomography chest scan. Pathological histology revealed lung adenocarcinoma in the patient. The UGP2-ALK mutation was found by next-generation sequencing. Subsequently, the patient was administered alectinib, and thereafter, the tumor lesion was observed to gradually shrink over the follow-up period. Progression-free survival reached 10 months as of the follow-up date, with no adverse events detected. This case report provides valuable insights into the clinical management of NSCLC patients with UGP2-ALK fusions. Moreover, alectinib is confirmed to be an appropriate therapeutic agent for such patients.

The clinicopathological features and renal prognostic factors in pure membranous lupus nephritis-a large series cohort study from China.

BACKGROUND: Membranous lupus nephritis (MLN) is a subepithelial immune deposition or its morphological sequelae with or without mesangial changes. Previous studies on the prognosis of MLN have shown relatively small sample sizes and short follow-up periods. METHODS: Our study was a retrospective analysis of biopsy-proven MLN patients from January 2010 to January 2020 at Jinling Hospital in China. The clinical manifestations, pathological features, and renal outcomes of MLN patients were collected. The endpoint was defined as end-stage kidney disease (eGFR≤15 mL/min·1.73 m2 or need for renal replacement therapy) or a doubling of serum creatinine or an eGFR decline of more than 40%. We used Cox regression to analyze the risk factors for renal outcome and Kaplan-Meier curves were used to analyze renal survival rate. RESULTS: In the total of 2884 lupus patients, we screened 535 MLN patients. 456 MLN patients were recruited with an average age of 34 ± 12 years, 87.8% for female patients and 62.1% patients of nephrotic syndrome with proteinuria of 2.67 g/24h. After follow-up of 78 (IQR, 47.3-113.0) months, 37 (8.1%) patients reached the renal endpoint. The 5-year and 10-year renal survival rates were 95.8% and 89.4%, respectively. 370 patients (81.1%) achieved complete remission, 43 patients (9.4%) had partial remission, and only 43 had no response. 34.4% MLN experienced a relapse. The Cox regression showed the risk factors that affect the renal prognosis include male, hypertension history, anemia, high uric acid, acute kidney injury, and interstitial fibrosis in the renal pathology. CONCLUSIONS: MLN mostly manifested as nephrotic syndrome, with few renal dysfunctions. Although MLN had a high relapse rate, most patients had a response to immunosuppressants and had a good renal outcome.

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.

Evaluation of four novel non-grain protein sources completely replacing soybean meal on growth performance, serum biochemistry, amino acid transport and intestinal health of grass carp (Ctenopharyngodon idella) at different water temperatures.

To investigate the effects of non-grain protein source and water temperature on growth and feed utilization differences of grass carp, the effects of different protein sources on the growth performance, serum biochemistry, digestive enzymes, amino acid transport and intestinal health of grass carp were studied at 24 °C, 28 °C and 32 °C. In this study, a total of 1350 grass carp (Ctenopharyngodon idella) (initial weight 5.00 ± 0.02 g) were selected, and Clostridium autoethanogenum protein (CAP), Tenebrio molitor meal (TMM), cottonseed protein concentrate (CPC) and Chlorella powder (CHP) were used as a single protein source to completely replace soybean meal for 56 days. The results showed that the final body weight (FBW), weight gain rate (WGR), specific growth rate (SGR) and protein efficiency ratio (PER) of grass carp increased significantly with the increasing temperature (P < 0.001). The CHP and SBM groups showed no significant differences in FBW, WGR, SGR and PER (P > 0.05), which were higher than the CAP, TMM and CPC groups (P < 0.001). The alanine transaminase (ALT), aspartate aminotransferase (AST), total protein (TP) and triglyceride (TG) concentrations of grass carp at 32 °C were significantly lower than those at 24 °C and 28 °C (P < 0.001). The acid phosphatase (ACP) activity decreased significantly with the increase of temperature (P = 0.001). The amylase (AMS) activity of the TMM, CPC and CHP groups was significantly lower than that of the SBM and CAP groups (P < 0.001), and the ACP and lipase (LPS) activities in the TMM group were significantly lower than those in the SBM group (P < 0.001). In addition, the interaction between temperatures and protein sources significantly affected the gene expression levels of amino acid transport including solute carrier family 1 member 3 (SLC1A3), solute carrier family 7 member 1 (SLC7A1), solute carrier family 7 member 5 (SLC7A5), solute carrier family 15 member 1b (SLC15A1b), solute carrier family 7 member 7 (SLC7A7), target of rapamycin (TOR), 4E binding protein 1 (4E-BP1) and ribosomal protein S6 kinase 1 (S6K1), intestinal inflammatory including tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-8 (IL-8), interleukin-10 (IL-10) and tight junction proteins (occludin, claudin1, claudin3, claudin7 and claudin11) (P ≤ 0.001). Collectively, our results indicated that CHP could be a potential protein source in the case of complete replacement of soybean meal in grass carp.

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.

Construction of novel π-bridged fluorescent probes for Fe2+ monitoring in living cells and foods.

Fe2+ plays a crucial role in biological systems as an essential trace element in the body. Iron is absorbed by the body through food and Fe2+ is also an important component of living cells; therefore, quantitative testing of Fe2+ in food and in living cells is of great importance. This paper presents the development of a novel π-bridge EDOT-based N-oxide turn-on fluorescent probe, designated as FeE, for the detection of Fe2+. The probe has been shown to exhibit excellent sensitivity, a favorable linear relationship between fluorescence signal intensity and Fe2+ concentration, and an effective immunity to interference. The probe has low cytotoxicity and has been successfully used to detect Fe2+ in cells using laser confocal fluorescence microscopy. It is also possible to determine the presence of Fe2+ in animal blood, spinach, apple juice, red wine, mineral water and metal cans.

Exploration of mitochondrial autophagy related genes in the diagnosis model construction and molecular marker mining of Alzheimer's disease based on multi-omics integration.

Key features of Alzheimer's disease include neuronal loss, accumulation of beta-amyloid plaques, and formation of neurofibrillary tangles. These changes are due in part to abnormal protein metabolism, particularly the accumulation of amyloid beta. Mitochondria are the energy production centers within cells and are also the main source of oxidative stress. In AD, mitochondrial function is impaired, leading to increased oxidative stress and the production of more reactive oxidative substances, further damaging cells. Mitophagy is an important mechanism for maintaining mitochondrial health, helping to clear damaged mitochondria, prevent the spread of oxidative stress, and reduce abnormal protein aggregation. To this end, this article conducts an integrated analysis based on DNA methylation and transcriptome data of AD. After taking the intersection of the genes where the differential methylation sites are located and the differential genes, machine learning methods were used to build an AD diagnostic model. This article screened five diagnostic genes ATG12, CSNK2A2, CSNK2B, MFN1 and PGAM5 and conducted experimental verification. The diagnostic genes discovered and the diagnostic model constructed in this article can provide reference for the development of clinical diagnostic models for AD.