AIM To investigate the effects of Congrong San on neuronal apoptosis and the Bax/Bcl-2/Caspase3 signaling pathway in a rat model of Alzheimer's disease(AD).METHODS A total of 60 2-month-old SD male rats were randomly divided into the blank group,the model group,the memantine hydrochloride group(0.025 g/kg)and low-dose,medium-dose and high-dose Congrong San groups(4.62,9.24,18.48 g/kg).All groups except the control group received stereotactic intracerebral injection of Aβ1-42 to establish AD models.Following the successful modeling,each group received its corresponding intragastric administration once daily for 28 consecutive days.After the administration,the rats had their learning and memory ability detected by the morris water maze test;their hippocampal neuronal morphology observed with HE and Nissl staining;their hippocampal neuronal apoptosis observed with TUNEL staining;and their hippocampal expressions of amyloid precursor protein(APP),β-site APP-cleaving enzyme 1(BACE1),and apoptosis-related proteins Bax,Bcl-2 and Caspase3 detected with Western blot assay.RESULTS Compared with the model group,the groups intervened with medium-dose and high-dose Congrong San exhibited improved learning and memory performance,alleviated hippocampal neuronal damage,increased Nissl body count(P＜0.01),reduced hippocampal apoptosis rate(P＜0.05,P＜0.01),decreased protein expressions of APP,BACE1,Bax and cleaved-Caspase3/Caspase3 ratio(P＜0.05,P＜0.01),and elevated Bcl-2 expression(P＜0.01).CONCLUSION Congrong San mitigates cognitive impairment,hippocampal neuronal damage,and apoptosis in AD rats,probably through inhibition of the Bax/Bcl-2/Caspase3 signaling pathway activation.

This study explores the mechanism of Guben Jiannao Liquid on Alzheimer's disease(AD) by regulating autophagy based on the liver kinase B1(LKB1)/adenosine monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR) pathway. Male SD rats were randomly divided into the blank group, model group, low-dose and high-dose groups of Guben Jiannao Liquid, and rapamycin group, with 10 rats in each group. Except for the blank group, all other groups of rats were injected bilaterally in the hippocampus with β-amyloid(Aβ)_(1-42) to establish the AD model. The low-dose(6.21 g·kg~(-1)) and high-dose(12.42 g·kg~(-1)) groups of Guben Jiannao Liquid and rapamycin group(1 mg·kg~(-1)) were given the corresponding drugs by gavage, and the blank and model groups were given an equal volume of saline by gavage for four weeks. Morris water maze was used to test the learning and memory ability of rats in each group; hematoxylin-eosin(HE) and Nissl staining were used to observe the morphological and quantitative changes of neurons and Nissl bodies in the CA1 region of rat hippocampus; immunohistochemistry was utilized to detect Aβ-positive cell expression in the CA1 region of rat hippocampus; transmission electron microscopy was employed to observe ultrastructural changes in rat hippocampal tissue, and Western blot was used to examine the protein expression levels of LKB1, p-AMPK/AMPK, p-mTOR/mTOR, Beclin1, p62, and LC3-Ⅱ in the hippocampal tissue of the rats. The results showed that compared with those in the blank group, rats in the model group had elevated evasion latency and decreased number of platform transversal and residence time in the platform quadrant. The number of neurons in the hippocampal area was reduced, and the morphology was impaired. The average integral optical density value of Aβ-positive cells was elevated; the expression levels of LKB1, p-AMPK/AMPK, Beclin1, and LC3-Ⅱ were decreased, and the expression levels of p-mTOR/mTOR and p62 were increased. Compared with those in the model group, rats in the low-dose and high-dose groups of Guben Jiannao Liquid had shorter evasion latency, higher number of platform transversal, longer residence time in the platform quadrant, increased number of neurons, decreased expression of Aβ-positive cells and average integral optical density values, and increased number of autophagic lysosomes in hippocampal tissue. The expression levels of LKB1, Beclin1, and LC3-Ⅱ were elevated in the hippocampus of rats in the low-dose group of Guben Jiannao Liquid. The expression levels of LKB1, p-AMPK/AMPK, Beclin1, and LC3-Ⅱ were elevated in the hippocampal tissue of rats in the high-dose group of Guben Jiannao Liquid, and the expression levels of p-mTOR/mTOR and p62 were decreased. The findings suggest that Guben Jiannao Liquid can improve cognitive impairment in AD rats, and its mechanism of action may be related to the activation of the LKB1/AMPK/mTOR signaling pathway and the up-regulation of autophagy level.
Animals ; Alzheimer Disease/physiopathology* ; Male ; TOR Serine-Threonine Kinases/genetics* ; Autophagy/drug effects* ; Rats, Sprague-Dawley ; Protein Serine-Threonine Kinases/genetics* ; AMP-Activated Protein Kinases/genetics* ; Rats ; Drugs, Chinese Herbal/administration & dosage* ; Signal Transduction/drug effects* ; AMP-Activated Protein Kinase Kinases ; Humans ; Hippocampus/metabolism*

This study aims to investigate the effect of Congrong San(CRS) on endoplasmic reticulum stress-induced neuroinflammation in the rat model of Aβ_(1-42)-induced Alzheimer's disease(AD). Sixty male Sprague-Dawley rats(2 months old) were randomized into blank(CON), model(MOD), low-dose Congrong San(L-CRS), medium-dose Congrong San(M-CRS), high-dose Congrong San(H-CRS), and memantine hydrochloride(MJG) groups. The Morris water maze test was carried out to examine the learning and memory abilities of rats in each group. Hematoxylin-eosin staining and Nissl staining were employed to observe the morphology and number of CA1 neurons in the hippocampus of rats in each group. The morphology and structure of the endoplasmic reticulum in the hippocampus were observed by transmission electron microscopy. The immunofluorescence assay was employed to detect the expression of 78 kDa glucose-regulated protein(GRP78) in the hippocampus. Western blot was employed to determine the expression of apoptosis-associated speck-like protein containing a CARD(ASC), cysteinyl aspartate-specific proteinase(caspase-1), interleukin-18(IL-18), interleukin-1β(IL-1β), GRP78, and pathway proteins including protein kinase RNA-like endoplasmic reticulum kinase(PERK), phosphorylated PERK(p-PERK), C/EBP homologous protein(CHOP), and NOD-like receptor pyrin domain-containing protein 3(NLRP3) in the rat hippocampus. Compared with the MOD group, the M-CRS and H-CRS groups showed improved learning and memory abilities, reduced neuron losses in the hippocampus, alleviated endoplasmic reticulum stress, inhibited PERK-CHOP-NLRP3 pathway, and lowered levels of IL-1β, IL-6, and tumor necrosis factor-alpha(TNF-α). The results suggest that CRS can alleviate cognitive impairment and hippocampal neuron damage and reduce neuroinflammation in AD rats by alleviating endoplasmic reticulum stress to inhibit the activation of NLRP3 inflammasomes.
Animals ; Endoplasmic Reticulum Stress/drug effects* ; Male ; Alzheimer Disease/psychology* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Rats, Sprague-Dawley ; Rats ; Inflammasomes/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Cognitive Dysfunction/metabolism* ; Disease Models, Animal ; Hippocampus/drug effects* ; Humans ; Neuroinflammatory Diseases/drug therapy*

Aim To explore the mechanism of Cong Rong San on AD model rats based on protein kinase R-like endoplasmic reticulum kinase(PERK)-eukaryotic initiation factor 2α(eIF2α)-nuclear factor kappa B(NF-κB)signaling pathway.Methods Sixty mice were randomly divided into normal group,model group,Cong Rong San groups(4.62,9.24,18.48 g·kg-1)and donepezil group,with 10 mice in each group.All groups of rats received bilateral hippocampal injections of Aβ1-42 to establish the AD model,except the normal group.After the intragastric administration,the Morris water maze behavior test was performed for rats to test-ed the learning and memory abilities.Nissl staining was detected the quantity and Nissl bodies of nerve cells.To detect the nuclear translocation of NF-κB by immu-nofluorescence.To observe the ultrastructure of endo-plasmic reticulum by Transmission electron microsco-py.ELISA for Aβ1-42 and inflammatory cytokines quantification.Western blot was used to detect the ex-pression level of protein in the hippocampus in PERK-eIF2α-NF-κB signaling pathway.Results The morris water maze results showed that Cong Rong San im-proved the escape latency time,increased the number of platform crossings,and prolonged the time spent in the target quadrant in AD rats.(P＜0.05 or P＜0.01).Nissl staining shows the neuronal cells are ar-ranged neatly,nucleus are present and the number of Nissl bodies was numerous and the number of neurons was increased in various doses of Cong Rong San.Im-munofluorescence showed that the expression of NF-κB in the nucleus of rats was decreased(P＜0.05 or P＜0.01).The shape of endoplasmic reticulum was neat,no significantly expanded,and the structure was normal in various doses of Cong Rong San.The levels of Aβ1-42,IL-1,TNF-α and the ratio of p-PERK/PERK,p-eIF2α/eIF2α,p-NF-κB p65/NF-κB p65 in hippo-campus of Cong Rong San group was significantly de-creased in ELISA and Western blot test(P＜0.05 or P＜0.01).Conclusion Cong Rong San can alleviates the immune inflammatory response of neuronal cells in the ERS state for improve the learning and memory a-bility of AD rats,the mechanism of action may through restraint the activation of PERK-eIF2α-NF-κB signa-ling pathway.

Aim To investigate the effects of Congrong San(CRS)on learning and memory ability,hippocam-pal neuronal injury,and ferroptosis in rats with Alzhei-mer's disease(AD)and to explore the related mecha-nisms.Methods AD rat models were established and divided into Sham,Model,CRS low-dose,CRS medium-dose,CRS high-dose,and memantine groups.After treatment,Morris water maze,HE and Nissl staining,transmission electron microscopy,immunofluorescence staining,Western blot,and kit assays were performed to assess learning and memory ability,hippocampal neuro-nal injury,ferroptosis-related indicatorsand glucose reg-ulated protein 78 ku(GRP78)-(proteinkinaseR-li-keERkinase)PERK-(activating transcription factor 4)ATF4 pathway protein expression.Results Com-pared with the model group,rats in the CRS medium-and high-dose groups and the memantine group showed significant improvement in learning and memory abili-ty,reduced hippocampal neuronal injury,increased number of Nissl bodies,and ameliorated endoplasmic reticulum swelling and mitochondrial damage.In addi-tion,the expressions of GRP78,p-PERK/PERK,and ATF4 were downregulated,while GPX4 expression was upregulated in the CRS medium-and high-dose groups and the memantine group.Moreover,MDA content de-creased,and SOD and GSH-PX levels increased in these groups.Conclusions CRS can improve the learning and memory ability in AD rats,reduce hipp-ocampal neuronal injury and ferroptosis,and its mecha-nism may be related to the inhibition of the GRP78-PERK-ATF4 pathway,enhancement of GPX4 expres-sion,and reduction of oxidative stress levels,providing a new approach for the clinical treatment of AD.

Aim To investigate the effects of Congrong San(CRS)on learning and memory ability,hippocam-pal neuronal injury,and ferroptosis in rats with Alzhei-mer's disease(AD)and to explore the related mecha-nisms.Methods AD rat models were established and divided into Sham,Model,CRS low-dose,CRS medium-dose,CRS high-dose,and memantine groups.After treatment,Morris water maze,HE and Nissl staining,transmission electron microscopy,immunofluorescence staining,Western blot,and kit assays were performed to assess learning and memory ability,hippocampal neuro-nal injury,ferroptosis-related indicatorsand glucose reg-ulated protein 78 ku(GRP78)-(proteinkinaseR-li-keERkinase)PERK-(activating transcription factor 4)ATF4 pathway protein expression.Results Com-pared with the model group,rats in the CRS medium-and high-dose groups and the memantine group showed significant improvement in learning and memory abili-ty,reduced hippocampal neuronal injury,increased number of Nissl bodies,and ameliorated endoplasmic reticulum swelling and mitochondrial damage.In addi-tion,the expressions of GRP78,p-PERK/PERK,and ATF4 were downregulated,while GPX4 expression was upregulated in the CRS medium-and high-dose groups and the memantine group.Moreover,MDA content de-creased,and SOD and GSH-PX levels increased in these groups.Conclusions CRS can improve the learning and memory ability in AD rats,reduce hipp-ocampal neuronal injury and ferroptosis,and its mecha-nism may be related to the inhibition of the GRP78-PERK-ATF4 pathway,enhancement of GPX4 expres-sion,and reduction of oxidative stress levels,providing a new approach for the clinical treatment of AD.

AIM To investigate the effects of Congrong San on neuronal apoptosis and the Bax/Bcl-2/Caspase3 signaling pathway in a rat model of Alzheimer's disease(AD).METHODS A total of 60 2-month-old SD male rats were randomly divided into the blank group,the model group,the memantine hydrochloride group(0.025 g/kg)and low-dose,medium-dose and high-dose Congrong San groups(4.62,9.24,18.48 g/kg).All groups except the control group received stereotactic intracerebral injection of Aβ1-42 to establish AD models.Following the successful modeling,each group received its corresponding intragastric administration once daily for 28 consecutive days.After the administration,the rats had their learning and memory ability detected by the morris water maze test;their hippocampal neuronal morphology observed with HE and Nissl staining;their hippocampal neuronal apoptosis observed with TUNEL staining;and their hippocampal expressions of amyloid precursor protein(APP),β-site APP-cleaving enzyme 1(BACE1),and apoptosis-related proteins Bax,Bcl-2 and Caspase3 detected with Western blot assay.RESULTS Compared with the model group,the groups intervened with medium-dose and high-dose Congrong San exhibited improved learning and memory performance,alleviated hippocampal neuronal damage,increased Nissl body count(P＜0.01),reduced hippocampal apoptosis rate(P＜0.05,P＜0.01),decreased protein expressions of APP,BACE1,Bax and cleaved-Caspase3/Caspase3 ratio(P＜0.05,P＜0.01),and elevated Bcl-2 expression(P＜0.01).CONCLUSION Congrong San mitigates cognitive impairment,hippocampal neuronal damage,and apoptosis in AD rats,probably through inhibition of the Bax/Bcl-2/Caspase3 signaling pathway activation.

Aim To explore the mechanism of Cong Rong San on AD model rats based on protein kinase R-like endoplasmic reticulum kinase(PERK)-eukaryotic initiation factor 2α(eIF2α)-nuclear factor kappa B(NF-κB)signaling pathway.Methods Sixty mice were randomly divided into normal group,model group,Cong Rong San groups(4.62,9.24,18.48 g·kg-1)and donepezil group,with 10 mice in each group.All groups of rats received bilateral hippocampal injections of Aβ1-42 to establish the AD model,except the normal group.After the intragastric administration,the Morris water maze behavior test was performed for rats to test-ed the learning and memory abilities.Nissl staining was detected the quantity and Nissl bodies of nerve cells.To detect the nuclear translocation of NF-κB by immu-nofluorescence.To observe the ultrastructure of endo-plasmic reticulum by Transmission electron microsco-py.ELISA for Aβ1-42 and inflammatory cytokines quantification.Western blot was used to detect the ex-pression level of protein in the hippocampus in PERK-eIF2α-NF-κB signaling pathway.Results The morris water maze results showed that Cong Rong San im-proved the escape latency time,increased the number of platform crossings,and prolonged the time spent in the target quadrant in AD rats.(P＜0.05 or P＜0.01).Nissl staining shows the neuronal cells are ar-ranged neatly,nucleus are present and the number of Nissl bodies was numerous and the number of neurons was increased in various doses of Cong Rong San.Im-munofluorescence showed that the expression of NF-κB in the nucleus of rats was decreased(P＜0.05 or P＜0.01).The shape of endoplasmic reticulum was neat,no significantly expanded,and the structure was normal in various doses of Cong Rong San.The levels of Aβ1-42,IL-1,TNF-α and the ratio of p-PERK/PERK,p-eIF2α/eIF2α,p-NF-κB p65/NF-κB p65 in hippo-campus of Cong Rong San group was significantly de-creased in ELISA and Western blot test(P＜0.05 or P＜0.01).Conclusion Cong Rong San can alleviates the immune inflammatory response of neuronal cells in the ERS state for improve the learning and memory a-bility of AD rats,the mechanism of action may through restraint the activation of PERK-eIF2α-NF-κB signa-ling pathway.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in healthcare facilities in major regions of China in 2023.Methods Clinical isolates collected from 73 hospitals across China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2023 Clinical & Laboratory Standards Institute (CLSI) breakpoints.Results A total of 445199 clinical isolates were collected in 2023,of which 29.0％ were gram-positive and 71.0％ were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species (excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi) (MRSA,MRSE and MRCNS) was 29.6％,81.9％ and 78.5％,respectively.Methicillin-resistant strains showed significantly higher resistance rates to most antimicrobial agents than methicillin-susceptible strains (MSSA,MSSE and MSCNS).Overall,92.9％ of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 91.4％ of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis had significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 93.1％ in the isolates from children and and 95.9％ in the isolates from adults.The resistance rate to carbapenems was lower than 15.0％ for most Enterobacterales species except for Klebsiella,22.5％ and 23.6％ of which were resistant to imipenem and meropenem,respectively .Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.6％ to 10.0％.The resistance rate to imipenem and meropenem was 21.9％ and 17.4％ for Pseudomonas aeruginosa,respectively,and 67.5％ and 68.1％ for Acinetobacter baumannii,respectively.Conclusions Increasing resistance to the commonly used antimicrobial agents is still observed in clinical bacterial isolates.However,the prevalence of important crabapenem-resistant organisms such as crabapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a slightly decreasing trend.This finding suggests that strengthening bacterial resistance surveillance and multidisciplinary linkage are important for preventing the occurrence and development of bacterial resistance.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in healthcare facilities in major regions of China in 2023.Methods Clinical isolates collected from 73 hospitals across China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2023 Clinical & Laboratory Standards Institute (CLSI) breakpoints.Results A total of 445199 clinical isolates were collected in 2023,of which 29.0％ were gram-positive and 71.0％ were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species (excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi) (MRSA,MRSE and MRCNS) was 29.6％,81.9％ and 78.5％,respectively.Methicillin-resistant strains showed significantly higher resistance rates to most antimicrobial agents than methicillin-susceptible strains (MSSA,MSSE and MSCNS).Overall,92.9％ of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 91.4％ of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis had significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 93.1％ in the isolates from children and and 95.9％ in the isolates from adults.The resistance rate to carbapenems was lower than 15.0％ for most Enterobacterales species except for Klebsiella,22.5％ and 23.6％ of which were resistant to imipenem and meropenem,respectively .Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.6％ to 10.0％.The resistance rate to imipenem and meropenem was 21.9％ and 17.4％ for Pseudomonas aeruginosa,respectively,and 67.5％ and 68.1％ for Acinetobacter baumannii,respectively.Conclusions Increasing resistance to the commonly used antimicrobial agents is still observed in clinical bacterial isolates.However,the prevalence of important crabapenem-resistant organisms such as crabapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a slightly decreasing trend.This finding suggests that strengthening bacterial resistance surveillance and multidisciplinary linkage are important for preventing the occurrence and development of bacterial resistance.