Open Field Experiment Research Articles

Protective effect and mechanism of Zuogui Jiangtang Jieyu Formula on damage to hippocampal synaptic microenvironment in rats with diabetes-related depression based on microglia-neuron crosstalk signal CD300f/TLR4

This study aims to reveal the protective effect and mechanism of Zuogui Jiangtang Jieyu Formula on the damage to hippo-campal synaptic microenvironment in rats with diabetes-related depression(DD) via regulating microglia immune receptor molecule-like family member f(CD300f)/Toll-like receptor 4(TLR4) signal. Firstly, the model of DD rats was established by a two-week high-fat diet+STZ injection+chronic mild and unpredictable stress plus isolation for 28 days. The rats were randomly divided into normal group, model group, CD300f blocker(CLM1, 2 μg·kg~(-1)) group, CD300f agonist(Fcγ, 5 μg·kg~(-1)) group, positive drug(0.18 g·kg~(-1) metformin+1.8 mg·kg~(-1) fluoxetine) group, and high-dose and low-dose(20.52 and 10.26 g·kg~(-1)) Zuogui Jiangtang Jieyu Formula groups. Depression-like behavior of rats was evaluated by open field and forced swimming experiments. The levels of blood glucose and insulin were detected by biochemical analysis. The levels of tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β), indoleamine 2, 3-dioxygenase(IDO), 5-hydroxytryptamine(5-HT), and dopamine(DA) in the hippocampus were detected by enzyme-linked immunosorbent assay. The changes in the synaptic ultrastructure in hippocampal neurons of rats were observed by transmission electron microscopy. The protein expressions of CD300f, TLR4, synaptophysin(SYN), and postsynaptic density protein 95(PSD-95) in microglial cells of the hippocampus were detected by immunofluorescence and Western blot. The results indicated that compared with that in the normal group, the total movement distance in open field experiments was reduced in the model group, and the immobility time in forced swimming experiments increased, with an elevated insulin level in serum, as well as TNF-α, IL-1β, and IDO levels in the hippocampus. The 5-HT and DA levels in the hippocampus were reduced. In addition, the CD300f expression was down-regulated in microglial cells of the hippocampus, and the TLR4 expression was up-regulated. Moreover, the expression of synapse-related proteins SYN and PSD-95 in hippocampal neurons decreased, and the synaptic ultrastructure of hippocampal neurons was significantly damaged. Compared with the model group, the CD300f blocker and agonist aggravated and alleviated the above abnormal changes, respectively. High-dose and low-dose Zuogui Jiangtang Jieyu Formula could significantly improve the above depression-like beha-vior in rats, inhibit the abnormal increase of TNF-α, IL-1β, and IDO and the decrease of 5-HT and DA, effectively increase the expression of CD300f in microglial cells, and decrease the expression of TLR4. They could up-regulate the protein expression of presyna-ptic membrane SYN and postsynaptic membrane PSD-95 in hippocampal neurons and finally improve the damage to the hippocampal synaptic microenvironment. In conclusion, this research confirmed that Zuogui Jiangtang Jieyu Formula effectively alleviated the depression-like behavior and inhibited inflammatory activation of microglial cells in the hippocampus of rats with DD, and the mechanism might be related to the regulation of CD300f/TLR4 signal to alleviate the damage to hippocampal synaptic microenvironment.

The Neuroprotective and Anxiolytic Effects of Magnesium Sulfate on Retinal Dopaminergic Neurons in 6-OHDA-Induced Parkinsonian Rats: A Pilot Study.

This study investigates the protective effects of magnesium sulfate on dopamine neurons in the retinas of rats with 6-hydroxydopamine (6-OHDA)-induced Parkinson's disease (PD). Rapidly progressing cognitive decline often precedes or coincides with the motor symptoms associated with PD. PD patients also frequently exhibit visual function abnormalities. However, the specific mechanisms underlying visual dysfunction in PD patients are not yet fully understood. Therefore, this study aims to investigate whether magnesium homeostasis affects dopaminergic neurons in the retina of PD rats. Thirty-six rats were divided into four groups: (1) control, (2) control with magnesium sulfate (control/MgSO4), (3) Parkinson's disease (PD), and (4) Parkinson's disease with magnesium sulfate (PD/MgSO4). The apomorphine-induced (APO) rotation test assessed the success of the PD models. The open-field experiment measured the rats' anxiety levels. Tyrosine hydroxylase (TH) and glutamate levels, indicators of dopamine neuron survival, were detected using immunofluorescence staining. Protein levels of solute carrier family 41 A1 (SCL41A1), magnesium transporter 1 (MagT1), and cyclin M2 (CNNM2) in the retina were analyzed using Western blot. Results showed that, compared to the PD group, rats in the PD/MgSO4 group had improved psychological states and motor performance at two and four weeks post-surgery. The PD/MgSO4 group also exhibited significantly higher TH fluorescence intensity in the left retinas and lower glutamate fluorescence intensity than the PD group. Additional experiments indicated that the protein levels of SLC41A1, MagT1, and CNNM2 were generally higher in the retinas of the PD/MgSO4 group, along with an increase in retinal magnesium ion content. This suggests that magnesium sulfate may reduce glutamate levels and protect dopamine neurons in the retina. Thus, magnesium sulfate might have therapeutic potential for visual functional impairments in PD patients.

Resveratrol by elevating the SIRT1 BDNF, GDNF and PSD95 levels reduce heroin addiction related behaviors

ObjectiveTo study the effects of resveratrol on heroin addiction-related behaviors and to preliminarily explore the possible intervention mechanism of resveratrol in heroin dependence. MethodsThe effects of resveratrol on heroin withdrawal symptoms were observed by naloxone; The effect of resveratrol on heroin reward memory acquisition was detected by CPP paradigm; The effect of resveratrol on the mental excitability of heroin was tested by open field experiment; The effect of resveratrol on heroin spatial learning and memory was tested by water maze test. Western blot was used to detect Sirtuin 1 (SIRT1) Expression of brain-derived neurotrophic factor (BDNF), glial cell derived neurotrophic factor (GDNF), and postsynaptic density protein (PSD95). ResultsThe behavioral results showed that the withdrawal behavior of the resveratrol intervention group was reduced compared with the heroin chronic dependence group (P<0.05), and the shift score of the conditioned place preference test of the resveratrol intervention group was reduced compared with the heroin chronic dependence group (P<0.05) The spatial learning and memory ability of the water maze in the resveratrol intervention group was improved compared with the heroin chronic dependence group (P<0.05), and the mental excitability of the resveratrol intervention group was lower than that of the heroin chronic dependence group (P<0.05), but higher than that of the saline group (P<0.05); SIRT1 The expression levels of BDNF, GDNF and PSD95 protein were significantly increased (P<0.05). ConclusionThe behavioral results of this study suggest that resveratrol can be used as a potential drug to treat heroin dependence. At the same time, SIRT1 The expression of BDNF, GDNF, and PSD95 increased; SIRT1, BDNF, GDNF, and PSD95 play an essential role in heroin addiction.

Degradation of Biodegradable Nonwoven Mulches in the Winter Period.

An open field experiment from November 2022 to May 2023 in Croatia, which is characterized by a continental humid climate, evaluated nonwoven mulches made from viscose, jute, and hemp fibres blended with PLA fibres. The blends of viscose and jute fibres (90:10, 80:20, and 70:30 ratios) were produced using mechanical web formation on cards with needle punching for bonding webs. Additionally, hemp fibres were blended with PLA fibres in a ratio of 80:20. Winter conditions caused significant structural changes in the mulches, including shrinkage, increased mass per unit area, thickness, and reduced air permeability. The amount of PLA fibre in the nonwoven mulch blends significantly affected nonwoven fabric structure change during exposure to winter conditions. After 180 days, the breaking force of all mulches increased by 30% to 277%. The soil beneath jute and hemp mulches maintained higher temperatures and moisture levels compared to viscose mulches. Soil organic carbon content varied with fibre type and was higher under jute and hemp mulches. K2O content was significantly higher in soils covered by mulches. All mulches effectively suppressed weeds. The experiment results showed that the newly produced nonwoven mulches could replace the conventional agro foil. Results also suggest that choosing biodegradable nonwoven mulches produced from fibres obtained from natural and renewable sources can influence soil fertility and the availability of nutrients, ultimately affecting plant growth and agricultural productivity.

Pubertal maternal presence reduces anxiety and increases adult neurogenesis in Kunming mice offspring

Puberty is a critical period of emotional development and neuroplasticity. However, most studies have focused on early development, with limited research on puberty, particularly the parental presence. In this study, four groups were established, and pubertal maternal presence (PMP) was assessed until postnatal days 21 (PD21), 28 (PD28), 35 (PD35), and 42 (PD42), respectively. The social interaction and anxiety behaviors, as well as the expression of oxytocin (OT) in the paraventricular nucleus (PVN) and supraoptic nucleus (SON), and the number of new generated neurons and the expression of estrogen receptor alpha (ERα) in the dentate gyrus (DG) were assessed. The results suggest that there is a lot of physical contact between the mother and offspring from 21 to 42 days of age, which reduces anxiety in both female and male offspring in adulthood; for example, the PMP increased the amount of time mice spent in the center area in the open field experiment and in the bright area in the light-dark box experiment. PMP increased OT expression in the PVN and SON and the number of newly generated neurons in the DG. However, there was a sexual difference in ERα, with ERα increasing in females but decreasing in males. In conclusion, PMP reduces the anxiety of offspring in adulthood, increases OT in the PVN and SON, and adult neurogenesis; ERα in the DG may be involved in this process.

Tangeretin alleviates inflammation and oxidative response induced by spinal cord injury by activating the Sesn2/Keap1/Nrf2 pathway.

Spinal cord injury (SCI) is a severe disabling disease that is characterized by inflammation and oxidative reactions. Tangeretin has been shown to possess significant antioxidant and anti-inflammatory activities. The Keap1/Nrf2 pathway, downstream of the Sesn2 gene, is involved in regulating the inflammation and oxidative response. The main objective of this study was to investigate the effect of tangeretin on SCI and its possible mechanism through cell and animal models. A T9 clamp injury was used for the mouse model and the LPS-induced stimulation of BV-2 cells was used for the cell model. The improvement of motor function after SCI was assessed by open field, swimming, and footprint experiments. The morphological characteristics of mouse spinal cord tissue and the levels of INOS, Sesn2, TNF-α, Keap1, Nrf2, IL-10, and reactive oxygen species (ROS) in vivo and in vitro were measured by several methods including western blotting, qPCR, immunofluorescence, HE, and Nissl staining. In vivo data showed that tangeretin can improve motor function recovery and reduce neuron loss and injury size in mice with SCI. Simultaneously, the in vitro findings suggested that treatment of BV-2 cells with tangeretin after LPS stimulation reduced the production of inflammatory factors and ROS, and could convert BV-2 cells from the M1 to the M2 type. Furthermore, Sesn2 knockout suppressed Keap1/Nrf2, inflammatory factors, ROS levels, and the M1 to M2 transition. Tangeretin can alleviate the inflammation and oxidative response induced by SCI by activating the Sesn2/Keap1/Nrf2 pathway.

Effects of transcranial magneto-acoustic stimulation on cognitive function and neural signal transmission in the hippocampal CA1 region of mice

As a new means of brain neuroregulation and research, transcranial magneto-acoustic stimulation (TMAS) uses the coupling effect of ultrasound and a static magnetic field to regulate neural activity in the corresponding brain areas. Calcium ions can promote the secretion of neurotransmitters and play a key role in the transmission of neural signals in brain cognition. In this study, to explore the effects of TMAS on cognitive function and neural signaling in the CA1 region of the hippocampus, TMAS was applied to male 2-month-old C57 mice with a magnetic field strength of 0.3 T and ultrasound intensity of 2.6 W/cm2. First, the efficiency of neural signaling in the CA1 region of the mouse hippocampus was detected by fiber photometry. Second, the effects of TMAS on cognitive function in mice were investigated through multiple behavioral experiments, including spatial learning and memory ability, anxiety and desire for novelty. The experimental results showed that TMAS could improve cognitive function in mice, and the efficiency of neural signaling in the CA1 area of the hippocampus was significantly increased during stimulation and maintained for one week after stimulation. In addition, the neural signaling efficiency in the CA1 area of the hippocampus increased in the open field (OF) experiment and recovered after one week, the neural signaling efficiency in the new object exploration (NOE) experiment was significantly enhanced, and the intensity slowed after one week. In conclusion, TMAS enhances cognitive performance and promotes neural signaling in the CA1 region of the mouse hippocampus.

Effects of water extract of the spleen-brain-related mineral drug Shehanshi on mouse sleep

ObjectiveTo investigate the effects of water extract of the spleen-brain-related mineral drug Shehanshi on mouse sleep. MethodsShehanshi water extract was subjected to component analysis via X-ray photoelectron spectroscopy. Then, the effects of low-dose (50 mg kg-1) and high-dose (100 mg kg-1) Shehanshi water extract on mouse sleep were evaluated through behavioral tests such as pentobarbital sodium subthreshold and above-threshold sleep experiments and autonomic activity experiments. Furthermore, transcriptome sequencing and nontarget metabolomics analysis were performed on the spleen and brain tissues of the mice. ResultsThe Shehanshi water extract contains a total of 30 elements and can reduce sleep latency, increase sleep time, and increase the sleep rate of mice. In the open field experiment, the movement distance of the mice decreased, and the central residence time and rest time increased. Immunoinfiltration analysis and immunohistochemical verification of spleen tissue showed that compared with those in the control group, the immune abundance of neutrophils in the administration groups increased (P < 0.05). Transcriptome data analysis revealed that the Atp1b2 gene was located at the intersection of the spleen and brain and was positively correlated with neutrophil expression but negatively correlated with the expression of the metabolite oleic acid in brain tissue. Immunohistochemical results showed that Atp2a3 protein expression decreased and Plcg1 protein expression increased in the high-dose group, and the difference was statistically significant (P < 0.05). The Atp1b2 protein level in the spleen tissue was positively correlated with that in the brain tissue of the mice (R = 0.829, P = 0.038). Western blotting revealed that Atp1b2 protein levels in the brain and spleen increased significantly in the high-dose group (P < 0.05). ConclusionThe mechanism by which Shehanshi water extract influences sleep may be associated with the expression of genes related to the spleen-brain axis and calcium signaling pathways in brain tissues.

Chilling at grain filling stage reduced rice grain protein content: An experimental and modeling study

Climate change has increased the trend in the intensity of global extreme weather events, including chilling. Nitrogen is one of the most essential nutrients for rice growth and development. Chilling will limit the uptake and translocation of nitrogen and the formation of grain protein in rice plants. The experiment conducted in the climate chamber with chilling stress applied at the grain filling stage revealed different effects on protein concentration in aboveground organs and caused asymmetry in grain protein contents (GPC) and grain protein yield (GPY). Chilling stress applied during the grain filling stage reduced the accumulation rate of rice grain protein by inhibiting the nitrogen uptake and the translocation from vegetative organs to grains, which resulted in a 35 % decrease in grain nitrogen accumulation and a 92 % increase in vegetative organs during grain filling. Consequently, the average nitrogen harvest index is reduced by 12 %. Nitrogen accumulation was severely affected when cooling degree days (CDD) ≥ 70 °C days. As the chilling intensity increased, the decrease of GPY was more significant than that of GPC. Moreover, we improved the grain temperature-nitrogen uptake function under chilling stress based on the relationship between CDD and the reduction in rice grains. By comparing the improved function and the modules in the existing crop model using the datasets from open field and artificial control experiments, we demonstrated that the current research on quantifying rice nitrogen uptake at extreme temperature stress needed further improvement. The effects of environmental stress on grain nitrogen accumulation are complex. Future studies should pay more attention to the ability of extreme temperature stress to affect nitrogen accumulation in various rice organs.

Cerebellar ferroptosis of hypertension mice following lead exposure

Exploring the changes in cerebellar ferroptosis in hypertensive mice after lead exposure. Twenty-five healthy C57 male mice were selected to construct a hypertensive model by intraperitoneal injection of angiotensin II(Ang II) at a concentration of 0.05 mg/kg for 7 consecutive days. After a systolic blood pressure of 140 mmHg, 20 hypertensive mice were randomly divided into a hypertensive control group and a hypertensive lead exposure group. Twenty C57 mice with normal blood pressure were randomly divided into a blood pressure normal control group and a blood pressure normal lead exposure group. The mice in the normal blood pressure control group and the hypertensive control group drank water freely. Mice in the lead exposure group with normal blood pressure and the lead exposure group with hypertension drank lead acetate water containing 250 mg/L. Ang II was injected intraperitoneally every two days in the hypertensive control group and hypertensive lead exposed group mice. Each group of mice was poisoned for 12 weeks. Using open field experiments and balance beam experiments to detect motor dysfunction in mice. Using a reagent kit to detect the levels of divalent iron(Fe~(2+)), malondialdehyde(MDA), and glutathione(GSH) in the cerebellum of different groups of mice. Western blot was used to determine the protein expression of member 11 of the solute carrier family 7(SLC7A11), glutathione peroxidase 4(GPX4), nuclear receptor coactivator 4(NCOA4), microtubule associated protein 1 light chain 3B(LC3B), and ferritin heavy chain 1(FTH1) in mouse cerebellar tissue. The result of the open field experiment showed that the activity distance(1013.04 cm) of mice in the hypertensive lead exposure group was significantly lower than that of the hypertensive control group(1351.18 cm) and the lead exposure group with normal blood pressure(1287.35 cm). And the lead exposure group with hypertension also extended the time through the balance beam, which was 29.40 seconds(P&lt;0.05). In addition, the Fe~(2+)content in the cerebellum of mice in the hypertensive lead exposure group was 3.33 μmol/g prot, which was 1.54 times that of the hypertensive control group and 1.14 times that of the lead exposure group with normal blood pressure. The MDA content was 4.71 nmol/mg prot, higher than that of the hypertensive control group and the lead exposure group with normal blood pressure. The GSH content was 5.36 μmol/g prot, lower than that of the hypertensive control group and the lead exposure group with normal blood pressure(P&lt;0.05). Western blot result showed that compared with the hypertensive control group and the lead exposure group with normal blood pressure, the protein expression of SLC7A11 and GPX4 in the hypertensive lead exposure group was significantly reduced(P&lt;0.05). In addition, compared with the control group with normal blood pressure, the expression of NCOA4 and LC3B proteins in the cerebellum of mice in the hypertension control group and lead exposure group with normal blood pressure increased, while the expression of FTH1 protein decreased(P&lt;0.05). The expression of NCOA4 and LC3B proteins in the hypertensive lead exposure group was higher than that in the hypertensive control group and the lead exposure group with normal blood pressure, while the expression of FTH1 protein decreased(P&lt;0.05). Lead exposure can exacerbate iron death in the cerebellar tissue of hypertensive mice, and iron autophagy may be involved in its occurrence and development.

Modeling Fragile X Syndrome: Characterizing Fmr1 Gene Knockout Mice across Genotype, Behavior, and Morphology

ABSTRACT Objective: Fragile X syndrome (FXS) is a hereditary condition resulting from dynamic mutations in the Fmr1 gene, leading to reduced or absent fragile X mental retardation protein (FMRP). Although molecular genetic diagnostics for FXS have advanced, there is currently a lack of effective clinical treatment. Hence, identifying a suitable FXS animal model is crucial for FXS therapeutic studies. This study primarily aims to evaluate whether Fmr1 mice display aberrant behaviors similar to those observed in human FXS patients, aiming to validate their feasibility as a model for FXS. Research Design and Methods: Polymerase chain reaction and immunoblotting were employed to genotype Fmr1 knockout (KO) mice and wild-type (WT) mice. Using hematoxylin and eosin staining and Golgi staining, testicular and dendritic spine morphology in both animal groups was observed. The animals were subjected to audiogenic seizure (AGS) observation, evaluating their learning and memory levels using the water maze and light avoidance methods, while their exploratory behavior capacity was assessed using the open-field and autonomous activity experiments. Anxiety behavior was evaluated using the elevated plus maze. Results: WT mice produced a 500 bp DNA fragment, while KO mice yielded an 800 bp fragment. KO mice exhibited a lack of FMRP expression, notably increased dendritic spine length and density, and presented with macroorchidism. KO mice showed increased susceptibility to AGS. In the Morris water maze experiment, KO mice displayed significantly increased escape latency and platform crossings. The spatial search experiment revealed that KO mice spent significantly less time in the target quadrant compared to WT mice. The light-avoidance and platform experiments indicated a significant reduction in latency for KO mice but an increase in errors. In the open-field experiment, KO mice spent significantly more time and made more entries into the central zone compared to WT mice. KO mice exhibited increased total travel distance and speed. The elevated plus maze experiment showed that KO mice spent significantly more time, made more entries, and traveled further in the open area compared to WT mice. The autonomous activity experiment revealed that KO mice had significantly more activity counts than WT mice but fewer standing counts. Conclusions: Findings from genotype, protein expression spectrum, morphology, and behavioral results suggest that Fmr1 gene KO mice demonstrate phenotypes similar to those observed in human FXS, making them a potential model for future FXS research. These phenotypes are associated with the absence of FMRP expression.

Establishment and evaluation of a premenstrual syndrome rat model induced by progesterone-withdrawal and emotional-stimulation

Premenstrual syndrome(PMS) lacks a highly consistent and feasible animal model that aligns with diagnostic and therapeutic standards in both traditional Chinese medicine(TCM) and western medicine, resulting in a lack of reliable experimental carriers for studying its pathogenesis and pharmacological effects. This study aims to systematically analyze the biological implications of PMS from the perspective of the &quot;disease-syndrome-symptom&quot; correlation and establish preparation and evaluation methods for an improved animal model of this disease. Firstly, clinical symptom gene sets related to the Qi stagnation syndromes due to liver depression and blood stasis in PMS in both modern medicine and TCM diagnostic standards were collected through GeneCards, DisGeNET, Mala-Cards, and the System of Foundational Diagnostic Association(SoFDA) database, as well as published literature. Based on the interaction information between genes, a &quot;disease-syndrome-symptom&quot; correlation network of PMS was established. Based on data mining results, an improved rat model of PMS was prepared by combining chronic restraint stress with the classical progesterone-withdrawal mo-del to simulate emotional depression caused by external environmental stimuli during the clinical onset process, inducing pathological damage from both physiological and emotional dimensions. The evaluation of the improved model before and after modification included open field experiment scores, organ indices, ovarian pathological changes, serum levels of estradiol(E_2), follicle-stimulating hormone/luteinizing hormone(FSH/LH), 5-hydroxytryptamine(5-HT), dopamine(DA), norepinephrine(NE), as well as coagulation parameters and hemorheology indexes. By calculating the degree, betweenness, and closeness centrality of nodes in the &quot;disease-syndrome-symptom&quot; correlation network, 163 core genes with topological importance were identified. Further biological function mining results indicated that core genes in PMS mainly participated in the regulation of the &quot;nervous-endocrine-immune&quot; system and pathways related to circulatory disorders. Mapping analysis of clinical phenotype symptom gene sets suggested significant correlations between core genes in PMS and depressive symptoms and pain symptoms caused by blood stasis. Compared with the simple progesterone withdrawal model, rats subjected to combined injections and restraint stress showed more significant abnormalities in open field experiment scores, ovarian tissue pathology, serum neurotransmitter levels of 5-HT and DA, as well as serum hormone levels of E_2 and FSH/LH. The modified modeling conditions exacerbated the pathological changes in blood rheology, coagulation function, and red blood cell morphology in model rats, confirming that the improved rat model could characterize the &quot;nervous-endocrine-immune&quot; system disorder and circulatory system disorders in the occurrence and progression of PMS, consistent with the clinical diagnostic and therapeutic standards of both TCM and western medicine. The establishment of the improved rat model of PMS can provide a reliable experimental carrier for elucidating the pathogenesis of PMS and discovering and evaluating therapeutic drugs. It also provides references for objectively reflecting the clinical characteristics of PMS in TCM and western medicine and precision treatment.

Polygonatum polysaccharide ameliorates D-galactose-induced cognitive dysfunction in aging rats by inhibiting ferroptosis through activation of Nrf2

ContextAging is a major risk factor for various neurodegenerative diseases, and ferroptosis has been identified as an important mode of cell death during accelerated aging. As the main component of the edible plant YuZhu in China, Polygonatum polysaccharide (POP) is an important natural compound with anti-aging properties. ObjectiveTo evaluate the anti-aging effects of POP and the underlying molecular mechanisms involved and to evaluate the overall anti-aging effects of POP on cognitive impairment due to accelerated aging. Materials and methodsA D-galactose (D-gal)-induced accelerated aging rat model was established to evaluate the anti-aging effects of POP and the underlying molecular mechanisms involved. In turn, Morris water maze and open field experiments were used to evaluate the anti-aging effects of POP on cognitive impairment due to accelerated aging. ResultsThe mechanism by which POP affects nuclear factor E2-related factor 2 (Nrf2), an essential transcription factor, was confirmed. POP significantly improved d-gal-induced cognitive dysfunction in treated model rats, which exhibited reduced pathological changes in the hippocampus, reduced latency of the water maze platform, and increased exploration time in the central area in the open field experiment compared to those of untreated model rats. Furthermore, POP intervention downregulated ferroptosis-related proteins and upregulated Nrf2 expression, and selective inhibition of Nrf2 eliminated the ability of POP to reduce ferroptosis. ConclusionsPOP is a natural ingredient with therapeutic potential due to its ability to alleviate aging by activating Nrf2, inhibiting ferroptosis, and alleviating cognitive dysfunction.

Explorations about the correlation between biological changes of meninges in periodontitis mice and cognitive impairment via single-cell RNA sequencing

Objective: To clarify the potential correlation between biological changes of meninges in periodontitis mice and cognitive impairment by analyzing the biological changes of meninges in periodontitis mice using single-cell RNA sequencing. Methods: Thirty C57BL/6 mice were divided into two groups by using random number table method (15 mice in each group). Mice in the control group were locally administered 2% carboxyl methyl cellulose (CMC) without Porphyromonas gingivalis (Pg) on both buccal sides. A mixture of Pg W83 and 2% CMC was applied on both buccal sides in the experimental group mice three times a week, lasting for 16 weeks in total. The absorption of alveolar bone, locomotor activity and cognitive function, the activation of microglia and astrocytes in the cortex were observed and assessed. The mRNA expression levels of Occludin in meninges and brain were detected in two groups. Single-cell RNA sequencing data of meninges were processed by uniform manifold approximation and projection (UMAP). Differential genes expressions of endothelial cells were processed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. In addition, real-time fluorescence quantitative PCR (RT-qPCR) was used to verify the expressions of transcription activating factor 3 (Atf3) and apolpoprotein L domain-containing 1 (Apold 1). Results: Methylene blue staining found the distances of buccal and palatal cement-enamel junction-alveolar bone crest in experimental mice [(185.60±17.60), (206.90±13.37) μm] increased significantly compared with the control group [(135.33±9.57), (163.05±14.98) μm] (t=5.02, P=0.002; t=4.37, P=0.005). Open field experiment showed the total distance and average speed of mice in the experimental group [(971.88±164.57) cm, (3.25±0.55) cm/s] were not statistically significant compared with the control group [(914.24±278.81) cm, (3.05±0.93) cm/s] (t=0.65, P=0.525; t=0.65, P=0.520). The recognition index of the experimental group [(48.02±16.92) %] was lower than the control group [(66.27±17.90) %] (t=2.40, P=0.027) by novel object recognition tests. Compared with the control group [(63.56±11.88) %], the alternation of experimental group [(50.99±14.17) %] was significantly decreased in Y maze tests (t=2.33, P=0.030). Immunohistochemistry results showed microglia and astrocytes were activated in the cortex of experimental mice. Compared with the control group (1.02±0.25, 1.04±0.31), the relative mRNA expressions of Occludin decreased significantly in the meninges and brain of periodontitis mice, respectively (0.61±0.10, 0.64±0.20) (t=3.47, P=0.010; t=2.66, P=0.024). By single-cell RNA sequencing, meninges cells were divided into 11 types, such as endothelial cells, fibroblasts, immune cells and so on. Endothelial cells were the main cell types in meninges [the control group: 26.47% (1 589/6 004), the experimental group: 26.26% (807/3 073)]. Compared with the control group [5.56% (334/6 004)], the percentage of granulocytes increased in the periodontitis mice [11.65% (358/3 073)]. Using clustering analysis to further focus on endothelial cells, GO enrichment analysis revealed differential genes were mainly related to angiogenesis, cell adhesion, apoptosis and so on. KEGG enrichment analysis revealed that differential genes were related to signaling pathways of interleukin-17, relaxin and so on. The relative mRNA expressions of Atf3 and Apold1 in meninges of periodontitis mice (0.42±0.24, 0.54±0.27) were significantly lower than the control group (1.03±0.26, 1.02±0.23) (t=3.88, P=0.005; t=3.02, P=0.017). Conclusions: The mice chronically infected with Pg W83 occurred memory impairment, neuroinflammation and changes of barrier function. In the meninges of periodontitis mice, there were infiltration of immune cells and down-regulation expressions of Atf3 and Apold1 by single-cell RNA sequencing. Meningeal immunity and changes of barrier function may play an important role in the cognitive impairment caused by periodontitis.

Regulating effect of Qifu Yin on intestinal microbiota in mice with memory impairment induced by scopolamine hydrobromide

Ethnopharmacological relevanceQifu Yin (QFY) originates from “Jingyue Quanshu · Volume 51 · New Fang Bazhen · Buzhen” a work by Zhang Jingyue, a distinguished Chinese medical practitioner from the Ming Dynasty. QFY is composed of Ginseng Radix et Rhizoma, Rehmanniae Radix Praeparata, Angelicae Sinensis Radix, Atractylodis Macrocephalae Rhizoma, Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle, Ziziphi Spinosae Semen, and Polygalae Radix. QFY is frequently employed to address memory loss and cognitive impairment stemming from vascular dementia, Alzheimer's disease (AD), and related conditions. Our findings indicate that QFY can mitigate nerve cell damage. Moreover, the study explores the impact of QFY on the calcium ion pathway and sphingolipid metabolism in mice with myocardial infarction, presenting a novel perspective on QFY's mechanism in ameliorating myocardial infarction through lipidomics. While this research provides an experimental foundation for the clinical application of QFY, a comprehensive and in-depth analysis of its improvement mechanism remains imperative. Aim of the studyTo clarify the regulatory mechanism of QFY on intestinal microecology in mice with memory impairment (MI). Material and methodsThe memory impairment mouse model was established by intraperitoneal injection of scopolamine hydrobromide. Kunming (KM) mice were randomly divided into blank group, Ginkgo tablet group (0.276 g/kg), QFY high, medium and low dose groups (17.2 g/kg, 8.6 g/kg, 4.3 g/kg). The effect on memory ability was evaluated by open field and step-down behavioral experiments. The morphological changes of nerve cells in the hippocampus of mice were observed by pathological method. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT) and glutathione peroxidase (GSH-Px) in the brain tissue of mice were detected. The expression levels of CREB, Brain-Derived Neurotrophic Factor (BDNF) and Recombinant Amyloid Precursor Protein (APP) in the hippocampus of mice were determined using immunohistochemistry. The expression of N-methyl-D-aspartate receptor (NMDAR) and cAMP response element binding protein (CREB) related factors in the serum of mice was analyzed by ELISA. The levels of apoptosis signal-regulating kinase-1 (ASK1) and c-Jun N-terminal kinase (JNK) mRNA in the hippocampus were detected by quantitative real-time fluorescence polymerase chain reaction (qPCR). The intestinal feces of mice were collected, and the 16 S rDNA technology was used to detect the changes in intestinal microbiota microecological structure of feces in each group. ResultsBehavioral experiments showed that the high-dose QFY group exhibited a significant increase in exercise time (P<0.05) and a decrease in diagonal time (P<0.05) compared to the model group. The medium-dose group of QFY showed a reduction in diagonal time (P<0.05). Additionally, the latency time significantly increased in the medium and high-dose groups of QFY (P<0.01). The number of errors in the low, medium and high dose groups was significantly decreased (P<0.05, P<0.01, P<0.01). The nerve cells in the CA1 and CA3 regions of QFY-treated mice demonstrated close arrangement and clear structure. Furthermore, the content of SOD significantly increased (P<0.01) and the content of MDA significantly decreased (P<0.05) in the low and high-dose QFY groups. The content of CAT in the medium-dose group significantly increased (P < 0.05). Immunohistochemical analysis showed a significant reduction in the number of APP expression particles in the CA1 and CA3 regions of all QFY groups. Moreover, BDNF expression significantly increased in the medium and high-dose groups, while CREB expression significantly increased in the low and medium-dose groups of QFY within the CA1 and CA3 regions. Serum analysis revealed significant increases in CREB content in the low, medium, and high dose groups of QFY (P<0.01, P<0.05, P<0.05), and decreases in NMDAR content across all QFY dose groups (P<0.01). PCR analysis showed a significant decrease in the contents of ASK1 and JNK in the medium-dose group (P<0.01). Microecological analysis of intestinal microbiota demonstrated a significant restoration trend in the relative abundance of Fusobacteria, Planctomycetes, and Verrucomicrobia (P<0.01 or P<0.05) at the phylum level in the QFY groups. At the genus level, Akkermansia, Paramuribaculum, Herminiimonas, Erysipelatoclostridium and other genera in the QFY groups showed a significant trend of relative abundance restoration (P<0.01 or P<0.05). ConclusionQFY can improve the memory of MI animals induced by scopolamine hydrobromide by restoring the homeostasis of intestinal microbiota and regulating related indexes in serum and brain tissue.

NMT2 alleviates depression-like behavior in a rat model of chronic unpredictable stress: An integrated proteomic and phosphoproteomic analysis

Proteomics has been widely used to investigate multiple diseases. Combining the analyses of proteomics with phosphoproteomics can be used to further explain the pathological mechanisms of depression. In this study, depression-like behavior was induced in a rat model of chronic unpredictable mild stress (CUMS). We subsequently conducted the sucrose preference test, open field experiment, and forced swimming test to assess depressive-like behavior. Proteomic and phosphoproteomic sequencing of the hippocampal tissues from depressive-like behavior and normal rats were analyzed to identify differentially expressed proteins (DEPs) and differentially phosphorylated proteins (DPPs). Differentially expressed phosphorylated proteins (DEPPs) were obtained by intersecting the DEPs and DPPs, and functional enrichment analysis, as well as ingenuity pathway analysis (IPA), were subsequently performed. The study also investigated correlations among the DEPPs and used qRT-PCR to quantify the expression levels of key genes. Five DEPPs were identified, Gys1, Nmt2, Lrp1, Bin1, and Atp1a1, which were found to activate the synaptogenesis signaling pathway, induce mitochondrial dysfunction, and activate the phosphoinositide biosynthesis and degradation pathways. The qRT-PCR results confirmed the proteomic findings for Gys1, Nmt2, Lrp1, and Atp1a1. Importantly, inhibiting Nmt2 was found to alleviate depression-like behavior and alleviate neuronal apoptosis in the hippocampus of CUMS rats. In conclusion, we identified five DEPPs associated with the synaptogenesis signaling pathway, mitochondrial dysfunction, and phosphoinositide biosynthesis and degradation in depression. Furthermore, NMT2 may be a potential target for the treatment or diagnosis of depression. Our findings provide novel insights into the molecular mechanisms of depression.

Assessment of Drought Tolerance of Two Cultivars of Indian Mustard (Brassica juncea) Using Morphological and Physiological Parameters

Aims: Drought stress may affect the Morphology, biochemistry, growth, and physiology of Brassica juncea (Indian mustard), a low-cost, oil-yielding, and economically significant plant. In this paper assessing various growth, morphological, and physiological parameters under different irrigation regimes could offer some significant information for the selection of a suitable and appropriate genotype for breeding. Study Design: Two commonly grown cultivars (RH-725 and RH-749) of mustard in Haryana under four irrigation regimes were evaluated. The applied Irrigation regimes were: control (double irrigation: once at the 50% flowering and another at the 50% fruiting stages), early irrigation (at 50% flowering only), late irrigation (at 50% fruiting only), and stress (no irrigation). The plants were exposed to short-term water shortages during the vegetative and reproductive growth stages. Drought stress in both stages had a negative effect on the morphological and physiological parameters of mustard. Place and Duration of Study: An open field experiment was performed at the Nursery of Kurukshetra University, Kurukshetra, Haryana, India, located at the latitude of 290–95° north and longitude of 760–82° east, with a height of 258.4 m above sea level. Crop sown under field conditions from October to March for two consecutive years (2018-2019, 2019-2020) at a temperature of 30-32 °C (days) and 15-25 °C (nights). Methodology: Measured adaptability using 15 morphological, 4 physiological, and 7 different growth parameters. PCA data revealed that physiological and growth parameters are more sensitive than morphological parameters in distinguishing the control and drought treatments. Alterations in physiological parameters occurred at all three levels of water stress. Growth analysis and yield parameters decreased with the severity of stress. Drought-stress treatments gradually reduced the Morphological and physiological traits of mustard. Results: The study revealed a relationship between Brassica species' adaptation to drought stress and the rapidity, severity, and duration of the drought event. This highlights the importance of considering these factors while selecting genotypes for breeding programmes aimed at improving drought tolerance in B. juncea. Overall, the research provides valuable information for selecting suitable and appropriate genotypes for breeding drought-tolerant varieties of B. juncea. By understanding the specific physiological (relative water content, leaf osmotic potential, membrane stability index, electrolytic index) and growth parameters (plant height, root length, fresh and dry weight of leaves and roots, leaf area etc.) that are more sensitive to drought stress. Furthermore, this study revealed a relationship between Brassica species' adaptation to drought stress and the rapidity, severity, and duration of the drought event. Based on PCA values, the cultivar RH-749 performed better both morphologically and physiologically under all stress conditions compared to RH-725. This suggests that RH-749 is more adapted and tolerant to drought stress, making it a potential candidate for breeding and cultivation in water-limited environments.

Inhibitory Effect of Evodiamine on Psoriasis Lesions and Itching in Mice.

This study seeks to investigate the effect of evodiamine on psoriasis and psoriatic pruritus. Imiquimod-induced psoriasiform dermatitis in mice was used as a model, and evodiamine was topically applied for seven days. The mice were observed daily for skin damage on the back, clinical score and their scratching behavior was recorded. Blood samples were collected on the final day of the experiment, and the serum levels of pruritus-associated inflammatory cytokines tumor necrosis factor (TNF) -α, interleukin (IL) -23, and IL-17A were measured using enzyme-linked immunosorbent assay. Histopathological changes were observed in Hematoxylin and Eosin-stained skin specimens. The expression levels of transient receptor potential vanilloid (TRPV) 1, TRPV3, TRPV4, and the pruritus-related mediators Substance P (SP), nerve growth factor (NGF), and calcitonin gene-related peptide (CGRP) in the skin lesions were analyzed using Western blot and qRT-PCR. The effect of evodiamine on the exploratory behavior, motor, and coordination abilities of mice was assessed using open field, suspension, and Rota-Rod experiments. Molecular docking was utilized to verify the binding of evodiamine to the residues of TRPV1, TRPV3, and TRPV4. Evodiamine reduced pruritus and inhibited inflammation by decreasing the levels of inflammatory mediators TNF-α, IL-23, and IL-17A in the serum of Imiquimod-induced mice and attenuated the mRNA and protein expression levels of SP, NGF, CGRP, TRPV1, TRPV3, and TRPV4 in the skin. Evodiamine is an effective treatment for psoriasis and pruritus, due to its ability to inhibit immune inflammation and pruritic mediators.

AI-Enabled Animal Behavior Analysis with High Usability: A Case Study on Open-Field Experiments

In recent years, with the rapid development of medicine, pathology, toxicology, and neuroscience technology, animal behavior research has become essential in modern life science research. However, the current mainstream commercial animal behavior recognition tools only provide a single behavior recognition method, limiting the expansion of algorithms and how researchers interact with experimental data. To address this issue, we propose an AI-enabled, highly usable platform for analyzing experimental animal behavior, which aims to provide better flexibility, scalability, and interactivity to make the platform more usable. Researchers can flexibly select or extend different behavior recognition algorithms for automated recognition of animal behaviors or experience more convenient human-computer interaction through natural language descriptions only. A case study at a medical laboratory where the platform was used to evaluate behavioral differences between sick and healthy animals demonstrated the high usability of the platform.

Harmonia axyridis (Boyer de Fonscolombe) (Hemiptera: Coccoidea) as a potential biological control agent of the invasive soft scale, Sphaerolecanium prunastri (Boyer de Fonscolombe) (Hemiptera: Coccoidea) in native wild apricot forests

BackgroundThe globose scale (GS), Sphaerolecanium prunastri (Boyer de Fonscolombe) (Hemiptera: Coccoidea), has invaded wild apricot forests in their native range in Central Eurasia threatening the ancestral germplasm resource. Biological control efficacy of the harlequin ladybird, Harmonia axyridis (Pallas, 1773) (Coleoptera: Coccinellidae) against the globose scale was assessed in laboratory and field experiments.ResultsIn the laboratory, Harmonia axyridis has a high feeding capacity on GS with numbers consumed daily increasing with temperature (15, 20, 25, 30, 35 °C), reaching an upper asymptote of 160–200 scales per day. In field cage experiments, efficacy of biological control (EBC) against first instar (49–99%) and second instar nymphs (20–80%) increased with GS density. When ants were present, control efficiency was reduced by 10–15%. In open-field experiments without cages, EBC was comparatively lower regardless of duration and how H. axyridis were released whether as adults, eggs cards or a mixture of adults and eggs cards.ConclusionsIn the long term, biological control with this ladybird predator could be considered as part of an IPM program package that includes banning or delaying mowing grass and understory plants in the forests that offer pollen and nectar for natural enemies.