Indirect Competitive ELISA for the Determination of Total Chromium Content in Food, Feed and Environmental Samples.

Background: This study aimed to prepare monoclonal antibodies (mAbs) with high immunoreactivity, sensitivity, and specificity for the chelate (Cr(III)-EDTA) of trivalent chromium ion (Cr(III)) and ethylenediamine tetraacetic acid (EDTA). Further, the study established an indirect competitive enzyme-linked immunosorbent assay (icELISA) for detecting the total chromium content in food, feed, and environmental samples. Methods: Hapten Cr(III)-iEDTA was synthesized by chelating Cr(III) with isothiocyanatebenzyl-EDTA (iEDTA). Immunogen Cr(III)-iEDTA-BSA formed by chelating Cr(III)-iEDTA with bovine serum albumin (BSA), and coating antigen Cr(III)-iEDTA-OVA formed by chelating Cr(III)-iEDTA with ovalbumin (OVA) were prepared using the isothiocyanate method and identified by ultraviolet spectra (UV) and inductively coupled plasma optical emission spectrometry (ICP-OES). Balb/c mice were immunized with the Cr(III)-iEDTA-BSA, and the anti Cr(III)-EDTA mAb cell lines were screened by cell fusion. The Cr(III)-EDTA mAbs were prepared by induced ascites in vivo, and their immunological characteristics were assessed. Results: The immunogen Cr(III)-iEDTA-BSA was successfully synthesized, and the molecular binding ratio of Cr(III) to BSA was 15.48:1. Three hybridoma cell lines 2A3, 2A11, and 3D9 were screened, among which 2A3 was the best cell line. The 2A3 secreted antibody was stable after six passages, the affinity constant (Ka) was 2.69 × 109 L/mol, its 50% inhibition concentration (IC50) of Cr(III)-EDTA was 8.64 µg/L, and it had no cross-reactivity (CR%) with other heavy metal ion chelates except for a slight CR with Fe(III)-EDTA (1.12%). An icELISA detection method for Cr(III)-EDTA was established, with a limit of detection (LOD) of 1.0 µg/L and a working range of 1.13 to 66.30 µg/L. The average spiked recovery intra-assay rates were 90% to 109.5%, while the average recovery inter-assay rates were 90.4% to 97.2%. The intra-and inter-assay coefficient of variations (CVs) were 11.5% to 12.6% and 11.1% to 12.7%, respectively. The preliminary application of the icELISA and the comparison with ICP-OES showed that the coincidence rate of the two methods was 100%, and the correlation coefficient was 0.987. Conclusions: The study successfully established an icELISA method that meets the requirements for detecting the Cr(III)-EDTA chelate content in food, feed, and environmental samples, based on Cr(III)-EDTA mAb, and carried out its preliminary practical application.

Morphophysiological responses of detached and adhered biofilms of Pseudomonas fluorescens to acidic electrolyzed water.

Pseudomonas spp. have emerged as the main spoilage bacteria, with many strains easily forming biofilms on food-contact surfaces and causing cross-contamination. The efficacy of disinfectants against bacteria is usually tested with planktonic cells; however, the disinfection tolerance of biofilms, especially detached biofilms, remains unknown. Here, we investigated the tolerance responses of detached and adhered biofilms of Pseudomonas fluorescens to acidic electrolyzed water (AEW) by determining tolerance responses by plate counting, comparing them using a Weibull model, and verifying changes in bacterial morphology by scanning electron microscopy. The experimental data and the responses calculated using Weibull a (scale) and b (shape) parameters agreed well (R2 values: 0.974-0.999), and we found that AEW exhibited effective antimicrobial activity against P. fluorescens, with adhered biofilms were more resistant than detached biofilms and planktonic cells. Additionally, AEW increased the bacterial membrane permeability and decreased the membrane potential, intracellular ATP concentrations, and intracellular pH while also triggering the disruption of extracellular polymeric substances. These results demonstrated that the morphophysiological responses of detached and adhered biofilms differed significantly and provided information on disinfectant-resistance strategies potentially beneficial to the development of novel disinfection approaches.

Alkyl polyglycoside and earthworm (Eisenia fetida) enhance biodegradation of green waste and its use for growing vegetables.

Managing municipal green waste is a challenge to municipalities, partly because of the slow rate of decomposition of green waste during composting due to its high lignin and cellulose contents. Hence, this study evaluated the effect of alkyl polyglycoside (APG), a biosurfactant, and the earthworm Eisenia fetida on the composting process. Addition of APG and E. fetida significantly increased total bacteria, cellulolytic fungi, phosphate solubilizing bacteria and nitrogen fixing bacteria populations, and the activities of cellulase, urease and alkaline phosphatase in composts as compared with the control. The APG and earthworm treatments also increased surface roughness and porosity of the green waste; Compared with control, APG and earthworm addition increased the degradation rate of TOC, lignin and cellulose by 5.9-17.9, 10.3-32.0 and 10.8-18.8%, respectively, and resulted in better compost quality, as was reflected in the neutral pH, higher cation exchange capacity (CEC) and nutrient concentrations (N, P, K, Ca, Mg, Fe, Cu, Zn, Mn). Final germination percentage and growth rate of tomato, eggplant and pepper seedlings were higher (P < 0.05) or similar in all composts produced with the addition of APG and earthworm, while plant growth was lower (P < 0.05) in the compost produced with the control than in peat substrate. The combination of APG+E. fetida enhanced the decomposition of green waste and improved final compost quality the most. Further research is needed to determine the best level of APG addition and optimum earthworm density for composting green waste.

Application of seasonal freeze-thaw to pretreat raw material for accelerating green waste composting.

The recalcitrance of green waste, caused by its high lignocellulose content, is a technical challenge for accelerating green waste composting. However, because lignocellulose degradation in litter (similar to green waste) can be promoted during the freeze-thaw season, and the composting is difficult to implement in this period (due to the low temperature); seasonal freeze-thaw was intended to be used as a pretreatment strategy for the existing technical challenge in the winter of cold regions. In this process, green waste was pretreated with seasonal freeze-thaw to enhance its lignocellulose degradation for subsequent composting. To verify this assumption, two strategies for the pretreatment were used: the green waste was either drenched or immersed in water during the freeze-thaw season, and the effects on subsequent composting were evaluated. The results demonstrated that both strategies can significantly promote the mineralization of TOC (total organic carbon, by 2.73%-8.01% compared with the control, the following comparisons were all based on the control), TN (total nitrogen, by 0.21%-0.52%), and lignocellulose (lignin degradation was promoted by 3.52%-3.73%, cellulose degradation was promoted by 13.23%-14.26%) during composting and that the synthesis of humus was also enhanced (by 19.19%-21.43%). Furthermore, since the loss of NH4+N and NO3-N was significantly less in the drenched treatment than in the immersed treatment (by 9.15% for the loss of NH4+N and 7.66% for the loss of NO3-N), drenching the green waste during the freeze-thaw season might be a better strategy than immersing for nitrogen conservation. An additional advantage of drenching compared to immersing is water conservation.

Bamboo biochar amendment improves the growth and reproduction of Eisenia fetida and the quality of green waste vermicompost.

Vermicomposting is a promising method for reusing urban green waste. However, high lignin content in the green waste could hinder the development of earthworm and microorganisms and the vermicomposting process, resulting in a low-quality vermicompost product. The objective of this study was to evaluate the effect of bamboo biochar addition (at 0%, 3%, and 6% on a dry w/w basis) on the activity of Eisenia fetida and the obtained vermicompost. Biochar addition increased (P < 0.05) earthworm biomass, juvenile and cocoon numbers of Eisenia fetida, as well as the activities of dehydrogenase, cellulase, urease and alkaline phosphatase. Compared to the control, lignin degradation rate was enhanced up to 13.89% by biochar addition. Biochar addition also improved the vermicompost quality in terms of cation exchange capacity (CEC), dissolved organic carbon (DOC) degradation, humification, nitrogen transformation, toxicity to germinating seeds (Brassica rapa L., Chinensis group) and heavy metals concentrations. The 6% bamboo biochar addition rate achieved maturity after 60 days of vermicomposting and resulted in the highest quality vermicompost based on parameters such as CEC, DOC, NH4+-N/NO3--N ratio, germination index and heavy metal concentration. We conclude that 6% biochar addition promoted earthworm growth and the vermicomposting of green waste.

The effect of anti-inflammatory properties of ferritin light chain on lipopolysaccharide-induced inflammatory response in murine macrophages.

Ferritin light chain (FTL) reduces the free iron concentration by forming ferritin complexes with ferritin heavy chain (FTH). Thus, FTL competes with the Fenton reaction by acting as an antioxidant. In the present study, we determined that FTL influences the lipopolysaccharide (LPS)-induced inflammatory response. FTL protein expression was regulated by LPS stimulation in RAW264.7 cells. To investigate the role of FTL in LPS-activated murine macrophages, we established stable FTL-expressing cells and used shRNA to silence FTL expression in RAW264.7 cells. Overexpression of FTL significantly decreased the LPS-induced production of tumor necrosis factor alpha (TNF-α), interleukin 1ß (IL-1ß), nitric oxide (NO) and prostaglandin E2 (PGE2). Additionally, overexpression of FTL decreased the LPS-induced increase of the intracellular labile iron pool (LIP) and reactive oxygen species (ROS). Moreover, FTL overexpression suppressed the LPS-induced activation of MAPKs and nuclear factor-κB (NF-κB). In contrast, knockdown of FTL by shRNA showed the reverse effects. Therefore, our results indicate that FTL plays an anti-inflammatory role in response to LPS in murine macrophages and may have therapeutic potential for treating inflammatory diseases.

Genetic manipulation of a high-affinity PHR1 target cis-element to improve phosphorous uptake in Oryza sativa L.

Phosphorus (P) is an essential macronutrient for crop development and production. Phosphate starvation response 1 (PHR1) acts as the central regulator for Pi-signaling and Pi-homeostasis in plants by binding to the cis-element PHR1 binding sequence (P1BS; GNATATNC). However, how phosphate starvation-induced gene expression is regulated remains obscure. In this work, we investigated the DNA binding affinity of the PHR1 ortholog OsPHR2 to its downstream target genes in Oryza sativa (rice). We confirmed that a combination of P1BS and P1BS-like motifs are essential for stable binding by OsPHR2. Furthermore, we report that variations in P1BS motif bases affected the binding affinity of OsPHR2 and that the highest affinity motif was GaATATtC (designated the A-T-type P1BS). We also found that a combination of two A-T-type P1BS elements in tandem, namely HA-P1BS, was very efficient for binding of OsPHR2. Using the cis-regulator HA-P1BS, we modified the promoters of Transporter Traffic Facilitator 1 (PHF1), a key factor controlling endoplasmic reticulum-exit of phosphate transporters to the plasma membrane, for efficient uptake of phosphorous in an energetically neutral way. Transgenic plants with the modified promoters showed significantly enhanced tolerance to low phosphate stress in both solution and soil conditions, which provides a new strategy for crop improvement to enhance tolerance of nutrient deficiency.

Comparison of nutritional profile between plant-based meat analogues and real meat: A review focusing on ingredients, nutrient contents, bioavailability, and health impacts.

In order to fully understand the nutritional heterogeneity of plant-based meat analogues and real meat, this review summarized their similarities and differences in terms of ingredients, nutrient contents, bioavailability and health impacts. Plant-based meat analogues have some similarities to real meat. However, plant-based meat analogues are lower in protein, cholesterol and VB12 but higher in dietary fiber, carbohydrates, sugar, salt and various food additives than real meat. Moreover, some nutrients in plant-based meat analogues, such as protein and iron, are less bioavailable. There is insufficient evidence that plant-based meat analogues are healthier, which may be related to the specific attributes of these products such as formulation and degree of processing. As things stand, it is necessary to provide comprehensive nutrition information on plant-based meat products so that consumers can make informed choices based on their nutritional needs.

Revealing extracellular protein profile and excavating spoilage-related proteases of Aeromonas salmonicida based on multi-omics investigation.

Aeromonas is a ubiquitous aquatic bacteria, and it is a significant factor contributing to meat spoilage during processing and consumption. The abilities of Aeromonas salmonicida 29 and 57, which exhibit spoilage heterogeneity, to secrete protease, lipase, hemolysin, gelatinase, amylase, and lecithinase were confirmed by plate method. A total of 3948 proteins were identified by ITRAQ in extracellular secretions of A. salmonicida, and 16 proteases were found to be potentially related to spoilage ability. The complete genome sequence of A. salmonicida 57 consists of one circular chromosome and three plasmids, while A. salmonicida 29 consists of one circular chromosome, without a plasmid. Transcriptomic analysis revealed a significant number of DEGs were up-regulated in A. salmonicida 29, which were mainly enriched in metabolic pathways (e.g., amino acid metabolism, carbohydrate metabolism), indicating that A. salmonicida 29 had better potential to decompose and utilize nutrients in meat. Six protease genes (2 pepB, hap, pepA, ftsI, and pepD) were excavated by combined ITRAQ with transcriptome analysis, which potentially contribute to bacterial spoilage ability and exhibit universality among other dominant spoilage bacteria. This investigation provides new insights and evidence for elucidating metabolic and spoilage phenotypic differences and provides candidate genes and strategies for future prevention and control technology development.

A trans-acting sRNA SaaS targeting hilD, cheA and csgA to inhibit biofilm formation of S. Enteritidis.

INTRODUCTION: Salmonella Enteritidis has brought great harm to public health, animal production and food safety worldwide. The biofilm formed by Salmonella Enteritidis plays a critical role in microbial cross-contamination. Small non-coding RNAs (sRNAs) have been demonstrated to be responsible for regulating the formation of biofilm. The sRNA SaaS has been identified previously, that promotes pathogenicity by regulating invasion and virulence factors. However, whether the SaaS is implicated in regulating biofilm formation in abiotic surfaces remains unclear. OBJECTIVES: This study aimed to clarify the effect of SaaS in Salmonella Enteritidis and explore the modulatory mechanism on the biofilm formation. METHODS: Motility characteristics and total biomass of biofilm of test strains were investigated by the phenotypes in three soft agar plates and crystal violet staining in polystyrene microplates. Studies of microscopic structure and extracellular polymeric substances (EPS) of biofilm on solid surfaces were carried out using confocal laser scanning microscope (CLSM) and Raman spectra. Transcriptomics and proteomics were applied to analyze the changes of gene expression and EPS component. The RNA-protein pull-down and promoter-reporter ß-galactosidase activity assays were employed to analyze RNA binding proteins and identify target mRNAs, respectively. RESULTS: SaaS inhibits biofilm formation by repressing the adhesion potential and the secretion of EPS components. Integration of transcriptomics and proteomics analysis revealed that SaaS strengthened the expression of the flagellar synthesis system and downregulated the expression of curli amyloid fibers. Furthermore, RNA-protein pull-down interactome datasets indicated that SaaS binds to Hfq (an RNA molecular chaperone protein, known as a host factor for phage Qbeta RNA replication) uniquely among 193 candidate proteins, and promoter-reporter ß-galactosidase activity assay confirmed target mRNAs including hilD, cheA, and csgA. CONCLUSION: SaaS inhibits the properties of bacterial mobility, perturbs the secretion of EPS, and contributes to the inhibition of biofilm formation by interacting with target mRNA (hilD, cheA, and csgA) through the Hfq-mediated pathway.

Traditional Chinese Medicine JianPiHuaTan formula improving quality of life and survival in patients with colorectal cancer through RAS/RAF downstream signaling pathways.

Objective: JianPiHuaTan Formula (JPHTF), a traditional Chinese medicine (TCM), has been utilized as an adjunctive therapy for colorectal cancer (CRC). The study aims to evaluate the potential clinical benefits of JPHTF and its effectiveness in inhibiting tumor growth. Methods: 300 stage II/III CRC patients and 412 advanced CRC patients were enrolled to verify the clinical value of JPHTF in CRC treatment. Furthermore, CRC patient-derived xenograft (PDX) mice were utilized to investigate the regulatory mechanisms of JPHTF. Results: JPHTF significantly improved abdominal distension, shortness of breath, drowsiness, loss of appetite, sleep, and tiredness in stage II/III CRC patients, thereby improving their quality of life. Simultaneously, JPHTF served as a supportive therapy in extending the overall survival (OS) of stage IV CRC patients with RAS/RAF mutations undergoing chemotherapy. Additionally, JPHTF effectively impeded tumor progression in CRC PDX models with RAS mutation, accompanied by a reduction in tumor cell content in the JPHTF group. Transcriptomic analysis revealed the involvement of the Hippo and Hedgehog signaling pathways in JPHTF-mediated CRC inhibition. Furthermore, mice in the JPHTF group exhibited increased immune cell infiltration. Conclusion: These findings suggested that JPHTF may inhibits tumor growth in CRC with RAS mutation by modulating RAS/RAF downstream signaling pathways, specifically the Hippo and Hedgehog signaling, leading to increased immune cell infiltration.

Influence paradigms of soil moisture on land surface energy partitioning under different climatic conditions.

Soil moisture (SM) directly controls the land surface energy partition which plays an important role in the formation of extreme weather events. However, its dependence on specific climatic conditions is not thoroughly understood due to the complexity of soil moisture effects. Here, we examine the relationship between SM and surface energy partitioning under different climate conditions, and identify the influence paradigms of soil moisture on surface energy partition. We find that temperature changes can explicitly determine the impact paradigm of different physical processes, i.e. evapotranspiration, soil freezing and thawing, and such influence paradigms are also affected by atmospheric aridity (VPD). Globally, there are five paradigms that effects on surface energy partitioning, including the warm-wet paradigm (WW), transitional paradigm (TP), warm-dry paradigm (WD), cool-wet paradigm (CW) and cold paradigm (CP). Since 1981, the global area proportion for TP is observed to increase pronouncedly. We also find that the critical SM threshold exhibits regional variations and the global average is 0.45 m3/m3. The identified paradigms and their long-term change trends provide new insights into the global intensification of land-atmosphere interaction, which has important implications for global warming and the formation of heatwaves.

Multiparametric bifurcations of an epidemiological model with strong Allee effect.

In this paper we completely study bifurcations of an epidemic model with five parameters introduced by Hilker et al. (Am Nat 173:72-88, 2009), which describes the joint interplay of a strong Allee effect and infectious diseases in a single population. Existence of multiple positive equilibria and all kinds of bifurcation are examined as well as related dynamical behavior. It is shown that the model undergoes a series of bifurcations such as saddle-node bifurcation, pitchfork bifurcation, Bogdanov-Takens bifurcation, degenerate Hopf bifurcation of codimension two and degenerate elliptic type Bogdanov-Takens bifurcation of codimension three. Respective bifurcation surfaces in five-dimensional parameter spaces and related dynamical behavior are obtained. These theoretical conclusions confirm their numerical simulations and conjectures by Hilker et al., and reveal some new bifurcation phenomena which are not observed in Hilker et al. (Am Nat 173:72-88, 2009). The rich and complicated dynamics exhibit that the model is very sensitive to parameter perturbations, which has important implications for disease control of endangered species.

SaaS sRNA promotes Salmonella intestinal invasion via modulating MAPK inflammatory pathway.

Salmonella Enteritidis is a foodborne enteric pathogen that infects humans and animals, utilizing complex survival strategies. Bacterial small RNA (sRNA) plays an important role in these strategies. However, the virulence regulatory network of S. Enteritidis remains largely incomplete and knowledge of gut virulence mechanisms of sRNAs is limited. Here, we characterized the function of a previously identified Salmonella adhesive-associated sRNA (SaaS) in the intestinal pathogenesis of S. Enteritidis. We found that SaaS promoted bacterial colonization in both cecum and colon of a BALB/c mouse model; it was preferentially expressed in colon. Moreover, our results showed that SaaS enhanced damage to mucosal barrier by affecting expressions of antimicrobial products, decreasing the number of goblet cells, suppressing mucin gene expression, and eventually reducing thickness of mucus layer; it further breached below physical barrier by strengthening invasion into epithelial cells in Caco-2 cell model as well as decreasing tight junction expressions. High throughput 16S rRNA gene sequencing revealed that SaaS also altered gut homeostasis by depleting beneficial gut microbiota while increasing harmful ones. Furthermore, by employing ELISA and western blot analysis, we demonstrated that SaaS regulated intestinal inflammation through sequential activation P38-JNK-ERK MAPK signaling pathway, which enabled immune escape at primary infection stage but strengthened pathogenesis at later stage, respectively. These findings suggest that SaaS plays an essential role in the virulence of S. Enteritidis and reveals its biological role in intestinal pathogenesis.

Plant-based meat analogues enhance the gastrointestinal motility function and appetite of mice by specific volatile compounds and peptides.

Eating behavior is critical for maintaining energy homeostasis. Previous studies have found that plant-based meat analogues increased diet intake in mice compared with animal meat under a free feeding mode, however the reasons were unclear. To explore the underlying mechanisms of plant-based meat analogues increasing diet intake, mice were fed animal or plant-based pork and beef analogue diets, respectively. Biochemical and histological analyses were performed to evaluate appetite-regulating hormones and gastrointestinal motility function. Peptiomics and GC-IMS were applied to identify key substances. We found that the intake of plant-based meat analogues significantly enhanced the gastrointestinal motility function of mice. The long-term intake (68 days) of plant-based meat analogues significantly increased the muscle layer thickness of the duodenum and jejunum of mice; the activity of gastrointestinal cells of Cajal were also promoted by upregulating the expression of c-kit related signals as compared to animal meat; plant-based meat analogues intake markedly enhanced the signal intensity of the intestinal neurotransmitter 5-hydroxytryptamine (5-HT) by upregulating the expression of 5-HT synthase and receptors but downregulating its transporter and catabolic enzyme in the intestine. Moreover, plant-based meat analogues intake significantly increased levels of appetite-stimulating factors in the peripheral or hypothalamus but reduced levels of appetite-suppressing factors compared with animal meat. Specific volatile compounds were significantly associated with appetite regulating factors. Among them, 7 substances such as linalool have a potential promoting effect on food intake. Besides, different digestive peptides in gastrointestinal tract may affect eating behavior mainly through the neuroactive ligand-receptor interaction pathway, exerting hormone-like effects or influencing endocrine cell secretion. These findings preliminarily clarified the mechanism of plant-based meat analogues promoting diet intake and provided a theoretical basis for a reasonable diet.

A Small RNA, SaaS, Promotes Salmonella Pathogenicity by Regulating Invasion, Intracellular Growth, and Virulence Factors.

Salmonella enterica serovar Enteritidis is a common foodborne pathogen that infects both humans and animals. The S. Enteritidis virulence regulation network remains largely incomplete, and knowledge regarding the specific virulence phenotype of small RNAs (sRNAs) is limited. Here, we investigated the role of a previously identified sRNA, Salmonella adhesive-associated sRNA (SaaS), in the virulence phenotype of S. Enteritidis by constructing mutant (ΔsaaS) and complemented (ΔsaaS/psaaS) strains. SaaS did not affect S. Enteritidis; it was activated in the simulated intestinal environment (SIE), regulating the expression of virulence target genes. We discovered that it directly binds ssaV mRNA. Caco-2 and RAW 264.7 cell assays revealed that SaaS promoted S. Enteritidis invasion and damage to epithelial cells while suppressing macrophage overgrowth and destruction. Furthermore, a BALB/c mouse model demonstrated that the deletion of SaaS significantly reduced mortality and attenuated the deterioration of pathophysiology, bacterial dissemination into systemic circulation, and systemic inflammation. Our findings indicate that SaaS is required for S. Enteritidis virulence and further highlight its biological role in bacterial pathogenesis. IMPORTANCE Salmonella is a zoonotic pathogen with high virulence worldwide, and sRNAs have recently been discovered to play important roles. We explored the biological characteristics of the sRNA SaaS and developed two cell infection models and a mouse infection model. SaaS is an SIE-responsive sRNA that regulates the expression of virulence-targeted genes. Additionally, it differentially mediates invasion and intracellular growth for survival and infection of the epithelium and macrophages. We further found that SaaS enhanced bacterial virulence by promoting lethality, colonization, and inflammatory response. These findings provide a better understanding of the critical role of sRNA in bacterial virulence.

[Effects of huannao yicong formula extract on behavior and ultrastructure of hippocampus mitochondria of APP transgenic mice of different months].

OBJECTIVE: To study the effects of Huannao Yicong formula (HNYCF) extract on behavior and ultrastructure of mitochondria in hippocampus CA1 area of APP transgenic mice of different months, and explore its partial mechanism in treating Alzheimer's disease (AD) through the perspective of energy metabolism. METHOD: One hundred and twenty APP695V717I transgenic mice of 3-month old were divided randomly into model group, Donepezil group (0.65 x 10(-3) g x kg(-1) x d(-1)), HNYCF extract large dose group (2.8 g x kg(-1) x d(-1)) and HNYCF extract small dose group (1.4 g x kg(-1) x d(-1)), and 30 mice in each group. Another 30 C57BL/6J mice with the same age and background were used as normal control group. All animals were administered once daily by gavage with the corresponding drug or distilled water. The course of intervention was 4 and 6 months. Behavioral changes were observed by Morris water maze test and step down test. Ultrastructure of mitochondria in hippocampus CA1 area was observed by transmission electron microscope. RESULT: At the age of 7 and 9 month, the number of times of passing through platform, swimming time and path length of model group increased significantly (P < 0.05, P < 0.01) in Morris water maze test, and the latent period decreased (P < 0.01) in step down test compared with normal group, and it would get worse with the development of disease course. HNYCF extract could increase the number of times of passing through platform, swimming time and path length (P < 0.05, P < 0.01) in Morris water maze test, prolong latent period in step down test of different age. At the age of 7 and 9 month, mitochondrial of hippocampus CA1 area was disrupted and dissolved. Most ridge structure arranged in a mess, and some ridge showed expanding, matrix loosing and swollen appearance, and it would get worse with the development of disease course. HNYCF extract could improve ultrastructure of mitochondria in hippocampus CA1 area, and increase its quality. CONCLUSION: Learning and memory ability decreased in APP transgenic mice model, and the quantity of neural mitochondria in hippocampus CA1 area with structure disrupting, and it would get worse with the development of disease course. HNYCF extract could improve the learning and memory ability of APP transgenic mice model, its mechanism might relate with improving ultrastructure of mitochondria in hippocampus, and increasing its quantity.

Effect of Physical Exercise Intervention Based on Improved Neural Network on College Students' Mental Health.

Physical exercise refers to various physical exercises carried out through certain means and methods. Physical exercise can not only achieve the purpose of strengthening the body and health but also make people face challenges in physical and mental sports, and the resulting psychological satisfaction and excitement make exercisers happy physically and mentally. Physical activity has a certain appeal that makes it one of the most effective ways for modern people to alleviate mental illnesses. The human brain's reaction time is linked to its thinking speed and intelligence. Regular physical activity can increase human brain cell reaction time and completely utilize human brain capacity, which is beneficial to the healthy development of human attention, memory, imagination, and thinking ability. Scientific and reasonable physical exercise is also an important means to improve people's intelligence levels. Everyone's physical health level is closely related to their mental health. A healthy mind comes from a healthy body. A large number of scientific studies have confirmed that reasonably arranging the contents and methods of physical exercise according to their own health level and psychological state cannot only enhance the physique of college students but also cultivate their excellent personality. Firstly, this paper summarizes the influence of physical exercise on mental health. The ways of employing physical exercise to improve and improve the mental health of college students are presented in this study. Then, this paper proposes a physical exercise intervention based on an improved neural network (NN), which has an impact on the mental health level of college students, and the effectiveness of this model is verified by simulation experiments.

Reconsidering Meat Intake and Human Health: A Review of Current Research.

Meat consumption is gradually increasing and its impact on health has attracted widespread attention, resulting in epidemiological studies proposing a reduction in meat and processed meat intake. This review briefly summarizes recent advances in understanding the effects of meat or processed meat on human health, as well as the underlying mechanisms. Meat consumption varies widely among individuals, populations, and regions, with higher consumption in developed countries than in developing countries. However, increasing meat consumption may not be the main cause of increasing incidence of chronic disease, since the development of chronic disease is a complex physiological process that involves many factors, including excessive total energy intake and changes in food digestion processes, gut microbiota composition, and liver metabolism. In comparison, unhealthy dietary habits and a sedentary lifestyle with decreasing energy expenditure are factors more worthy of reflection. Meat and meat products provide high-value protein and many key essential micronutrients. In short, as long as excessive intake and overprocessing of meats are avoided, meat remains an indispensable source of nutrition for human health.

Predictive Models for Knee Pain in Middle-Aged and Elderly Individuals Based on Machine Learning Methods.

Aim: This study used machine learning methods to develop a prediction model for knee pain in middle-aged and elderly individuals. Methods: A total of 5386 individuals above 45 years old were obtained from the National Health and Nutrition Examination Survey. Participants were randomly divided into a training set and a test set at a 7 : 3 ratio. The training set was used to create a prediction model, whereas the test set was used to validate the proposed model. We constructed multiple predictive models based on three machine learning methods: logistic regression, random forest, and Extreme Gradient Boosting. The model performance was evaluated by areas under the receiver (AUC), sensitivity, specificity, positive predictive value, and negative predictive value. Additionally, we created a simplified nomogram based on logistic regression for better clinical application. Results: About 31.4% (1690) individuals were with self-reported knee pain. The logistic regression showed that female gender (odds ratio [OR] = 1.28), pain elsewhere (OR = 4.64), and body mass index (OR = 1.05) were significantly associated with increased risk of knee pain. In the test set, the logistic regression (AUC = 0.71) showed similar but slightly higher accuracy than the random forest (AUC = 0.70), while the performance of the Extreme Gradient Boosting model was less reliable (AUC = 0.59). Based on mean decrease accuracy, the most important first five predictions were pain elsewhere, waist circumference, body mass index, age, and gender. Additionally, the most important first five predictions with the highest mean decrease Gini index were pain elsewhere, body mass index, waist circumference, triglycerides, and age. The nomogram model showed good discrimination ability with an AUC of 0.75 (0.73-0.77), a sensitivity of 0.72, specificity of 0.71, a positive predictive value of 0.45, and a negative predictive value of 0.88. Conclusion: This study proposed a convenient nomogram tool to evaluate the risk of knee pain for the middle-aged and elderly US population in primary care. All the input variables can be easily obtained in a clinical setting, and no additional radiologic assessments were required.