OBJECTIVES: To explore the effects of cannabidiol on endoplasmic reticulum stress and neuronal apoptosis in rats with multiple concussions (MCC). METHODS: SD rats were randomized into sham group, MCC group, 1% tween20 (TW) treatment group, and low-dose (10 mg/kg) and high-dose (40 mg/kg) cannabidiol treatment groups. In all but the sham group, MCC models were established using a metal pendulum percussion device, after which the rats received daily intraperitoneal injections of the corresponding agents for 2 weeks. The expressions of PERK, eIF2α, ATF4, CHOP, TRIB3, p-Akt and pro-caspase-3 in the brain tissue of the rats were detected with qRT-PCR, Western blotting and immunofluorescence staining. The core targets of cannabidiol in treatment of traumatic brain injury (TBI) were identified by network pharmacology analysis, and molecular docking was carried out to simulate the interaction of cannabidiol with the factors related to endoplasmic reticulum stress and apoptosis. RESULTS: Compared with the sham-operated rats, the rat models of MCC showed significantly increased mRNA expressions of PERK, eIF2α and CHOP and protein expressions of PERK, eIF2α, ATF4, CHOP, TRIB3, p-AKT and pro-caspase-3 in the cerebral cortex. CBD treatment, especially at the high dose, obviously increased the expression of p-Akt and lowered the expression levels of the other factors tested in the rat models. Network pharmacology analysis indicated interactions of the core targets of CBD with the factors related to endoplasmic reticulum stress and TBI, and molecular docking study showed a high binding energy of CBD with multiple factors pertaining to endoplasmic reticulum stress and apoptosis. CONCLUSIONS MCC induce endoplasmic reticulum stress and apoptosis in rat brain tissues, for which CBD, especially at a high dose, provides neuroprotective effects by inhibiting endoplasmic reticulum stress and cell apoptosis.
Animals ; Endoplasmic Reticulum Stress/drug effects* ; Apoptosis/drug effects* ; Rats, Sprague-Dawley ; Activating Transcription Factor 4/metabolism* ; Transcription Factor CHOP/metabolism* ; Rats ; Eukaryotic Initiation Factor-2/metabolism* ; Signal Transduction/drug effects* ; eIF-2 Kinase/metabolism* ; Cannabidiol/pharmacology* ; Neurons/metabolism* ; Brain Concussion/metabolism* ; Male ; Molecular Docking Simulation

AIM:To determine if scutellarin(Scu)provides neuroprotection by reducing neuronal apoptosis in rats subjected to middle cerebral artery occlusion(MCAO)via the inhibition of the JAK2/STAT3 signalling pathway.METHODS:Proteins linked to Scu and ischaemic stroke-induced neuronal apoptosis were identified using the Swiss Tar-get Prediction,PharmMapper,OMIM,and GeneCards databases.Intersecting targets were identified through Venn analy-sis.Protein-protein interaction networks were visualised utilising Cytoscape software,and principal targets were identi-fied.Enrichment analyses of GO functions and KEGG pathways were conducted utilising the Metascape database.Molecu-lar docking of Scu with core targets was performed utilising AutoDock Vina.The neuroprotective effects of Scu were as-sessed in MCAO rats using Zea Longa scoring and the suspension test.JAK2/STAT3 phosphorylation levels and apoptosis-related proteins[cleaved caspase-3(C-caspase-3),caspase-3,Bax,and Bcl-2]were assessed using Western blot and im-munofluorescence staining.The JAK2-specific inhibitor AG490 was employed to further investigate the role of the JAK2/STAT3 signaling pathway.RESULTS:Network pharmacology analysis revealed 832 shared targets,with pathways en-riched in tumor-associated pathways,the JAK/STAT signalling pathway,and the HIF-1 signalling pathway.Molecular docking revealed robust binding affinities of Scu with the ten principal targets.Behavioural assessments utilising Zea Lon-ga scoring and the suspension test demonstrated that Scu markedly enhanced neurological recovery in MCAO rats.Western blot and immunofluorescence analyses demonstrated that phosphorylation levels of JAK2 and STAT3,along with the ex-pression of C-caspase-3,Bax,and Bcl-2,were markedly elevated in the MCAO group relative to the sham group(P<0.05).Post Scu treatment,phosphorylation levels of JAK2 and STAT3,along with C-caspase-3 and Bax expression,were markedly diminished,whereas Bcl-2 expression and fluorescence intensity were substantially increased(P<0.05).In the combined AG490 and Scu treatment group(MCAO+Scu+AG490),the phosphorylation levels of JAK2 and STAT3,as well as the expression of C-caspase-3 and Bax,exhibited no significant difference when compared to the Scu-alone group(P>0.05).Bcl-2 expression and fluorescence intensity were markedly reduced in the combined AG490 and Scu treatment group relative to the Scu-alone group(P<0.05).CONCLUSION:Scu seems to provide neuroprotection in ischaemic stroke by reducing neuronal apoptosis through the inhibition of the JAK2/STAT3 signalling pathway.

AIM:To determine if scutellarin(Scu)provides neuroprotection by reducing neuronal apoptosis in rats subjected to middle cerebral artery occlusion(MCAO)via the inhibition of the JAK2/STAT3 signalling pathway.METHODS:Proteins linked to Scu and ischaemic stroke-induced neuronal apoptosis were identified using the Swiss Tar-get Prediction,PharmMapper,OMIM,and GeneCards databases.Intersecting targets were identified through Venn analy-sis.Protein-protein interaction networks were visualised utilising Cytoscape software,and principal targets were identi-fied.Enrichment analyses of GO functions and KEGG pathways were conducted utilising the Metascape database.Molecu-lar docking of Scu with core targets was performed utilising AutoDock Vina.The neuroprotective effects of Scu were as-sessed in MCAO rats using Zea Longa scoring and the suspension test.JAK2/STAT3 phosphorylation levels and apoptosis-related proteins[cleaved caspase-3(C-caspase-3),caspase-3,Bax,and Bcl-2]were assessed using Western blot and im-munofluorescence staining.The JAK2-specific inhibitor AG490 was employed to further investigate the role of the JAK2/STAT3 signaling pathway.RESULTS:Network pharmacology analysis revealed 832 shared targets,with pathways en-riched in tumor-associated pathways,the JAK/STAT signalling pathway,and the HIF-1 signalling pathway.Molecular docking revealed robust binding affinities of Scu with the ten principal targets.Behavioural assessments utilising Zea Lon-ga scoring and the suspension test demonstrated that Scu markedly enhanced neurological recovery in MCAO rats.Western blot and immunofluorescence analyses demonstrated that phosphorylation levels of JAK2 and STAT3,along with the ex-pression of C-caspase-3,Bax,and Bcl-2,were markedly elevated in the MCAO group relative to the sham group(P<0.05).Post Scu treatment,phosphorylation levels of JAK2 and STAT3,along with C-caspase-3 and Bax expression,were markedly diminished,whereas Bcl-2 expression and fluorescence intensity were substantially increased(P<0.05).In the combined AG490 and Scu treatment group(MCAO+Scu+AG490),the phosphorylation levels of JAK2 and STAT3,as well as the expression of C-caspase-3 and Bax,exhibited no significant difference when compared to the Scu-alone group(P>0.05).Bcl-2 expression and fluorescence intensity were markedly reduced in the combined AG490 and Scu treatment group relative to the Scu-alone group(P<0.05).CONCLUSION:Scu seems to provide neuroprotection in ischaemic stroke by reducing neuronal apoptosis through the inhibition of the JAK2/STAT3 signalling pathway.

Background Workers engaged in benzene-exposed or benzene-containing solvent-exposed operations in China are predominantly subjected to a low concentration of benzene series compounds, and prolonged exposure to low concentrations of benzene, toluene, and xylene (BTX) may have implications for blood pressure. Objective To investigate the influence of low-concentration BTX exposure on the blood pressure of workers, aiming to provide a basis for enterprises to devise associated health management strategies to mitigate the occurrence of hypertension among workers exposed to low concentrations of BTX. Methods Using a cross-sectional design, 884 workers from a petroleum refining enterprise in Hainan who participated in an occupational health examination in 2022 were selected as the study population, and were divided into an exposure group of 649 workers and a control group of 235 workers based on their reporting of BTX exposure or not. Data on workplace BTX concentrations and health examinations of the study subjects were collected and questionnaires were administered. In addition, S-phenylmercapturic acid (S-PMA), hippuric acid (HA), and methyl hippuric acid (MHA, including the three isomers 2-MHA, 3-MHA, and 4-MHA) were measured in the urine of the workers using high-performance liquid chromatography-tandem triple quadrupole mass spectrometry to assess internal BTX burden. The effects of low-concentration BTX exposure on blood pressure were analyzed. Results In 2022, the concentrations of benzene, toluene, and xylene of all monitoring points did not exceeded the national limits by either time-weighted average (TWA) or short-term exposure limit (STEL), indicating low-concentration BTX exposure. Regarding the internal burden of BTX, the concentrations of benzene metabolite S-PMA, toluene metabolite HA, and xylene metabolites 3-MHA and 4-MHA in the urine samples in the exposure group were higher than those in the control group (P < 0.05). The correlation analysis showed a positive correlation between urinary S-PMA concentration and diastolic blood pressure in the workers (r=0.265, P < 0.05). Differences in systolic and diastolic blood pressure distributions were statistically significant among workers grouped by sex, age, work years, educational levels, monthly income, body mass index (BMI), alcohol use, dietary oil, and types of residential address (P < 0.05). Significant differences in systolic blood pressure distribution were observed among workers by smoking status and levels of labor intensity (P < 0.05). Compared with the control group, the workers in the exposure group exhibited a significant increase in diastolic blood pressure (P < 0.05). The results of multiple linear regression showed that age, sex, and BMI had statistically significant effects on systolic blood pressure (P < 0.05), while age, work years, and BMI had statistically significant effects on diastolic blood pressure (P < 0.05). The systolic blood pressure of age > 35 years, male, overweight and obese workers was significantly higher than that of age ≤ 35 years, female, and underweight workers, and the diastolic blood pressure of age > 35 years, work years > 5 years, and obese workers was higher than age ≤35 years, ≤5 years of service, and underweight workers. Low-concentration BTX exposure was one of the main influencing factors for elevated diastolic blood pressure, and the exposed workers showed a 1.337 mmHg increase in diastolic blood pressure compared to the control group (P < 0.05). Conclusion Low-concentration BTX exposure, work years > 5 years, and obesity may elevate blood pressure among petroleum refininig workers. Regular blood pressure monitoring and enhanced health interventions for this occupational group are warranted.

Objective To analyze the current status and influencing factors of elevated blood pressure among employees in a large petroleum refining enterprise in Hainan Province. Methods A total of 940 workers from a petroleum refining enterprise in Hainan Province was selected as the study subjects using the convenience sampling method. The results of their health status survey, occupational medical examination, and occupational stress measurement were collected. Results The detection rate of elevated blood pressure in the study subjects was 23.9% (225/940), with the detection rate of normal blood pressure and hypertension of 17.7% (166/940) and 6.3% (59/940), respectively. The detection rate of occupational stress was 28.8% (271/940). The result of multivariate logistic regression analysis showed that workers aged 30 -<40, 40 -<50, and ≥50 years had a higher risk of elevated blood pressure than those aged <30 years after controlling for confounding factors such as gender, residential address, length of service, education level, personal monthly income, smoking status, physical exercise, salt intake, oil intake, occupational stress, and high temperature exposure (all P<0.05). Workers in the body mass index (BMI) overweight group and obese group had a higher risk of elevated blood pressure than those in the normal group (all P<0.05). The risk of elevated blood pressure was higher in workers who drinks than those who did not (P<0.05). Workers exposed to noise levels of 85-90 dB(A) had a lower risk of elevated blood pressure compared to those exposed to noise levels >90 dB(A) (P<0.05). Conclusion Age, BMI, drinking status, and noise exposure levels are independent influencing factors for elevated blood pressure among workers in this petroleum refining enterprise. Blood pressure management should be strengthened for workers aged ≥30 years, overweight, obesity, alcohol consumption and with noise exposure intensity > 90 dB(A).

Objective:Investigating the effect of scutellarin on the expression of silent information regulator 1/nucle-ar factor κB(SIRT1/NF-κB)signaling pathway in microglia of cerebral ischemic rat.Methods:Male SD rats were de-vided into sham group,middle cerebral artery occlusion group(MCAO group),MCAO+scutellarin treatment group(MCAO+S group)randomly.Western Blot and immunofluorescence were used to detect the expression of SIRT1 and NF-κB(p65)and phosphorylation of NF-κB(p-p65)of ischemic cortex of rats.Results:Results of Western Blot and immunofluorescence showed that scutellarin pretreatment significantly increased the expression of SIRT1 and reduced phosphorylation of NF-κB p65(P＜0.05).Conclusion:Scutellarin could regulate SIRT1/NF-κB signaling pathway in microglia of cerebral ischemic rat.

Objective:To investigate the effects of cannabidiol(CBD)on the NOD-like receptor protein 3(NLRP3)inflammasome in the brains of rats with multiple cerebral concussions(MCC).Methods:Rats were subjec-ted to the MCC model and divided into Sham,MCC,vehicle(MCC+TW),CBD-L(10 mg/kg),and CBD-H(40 mg/kg)groups.Immunofluorescence double staining was used to observe changes in NLRP3 and microglial cells in the brain,and Western Blot was performed to detect the expression changes of the NLRP3 inflammasome.Results:A sig-nificant increase in lectin-positive microglial cells of the cortex with enlarged cell bodies and elevated immunofluores-cence intensity of NLRP3 in the activated microglial cells was revealed by immunofluorescence double staining following MCC(P＜0.05).The immunofluorescence intensity of NLRP3 in the activated microglial cells was downregulated by the administration of CBD,with a more pronounced effect observed in the CBD-H group compared to the CBD-L group(P＜0.05).The expression of NLRP3,caspase-1,and apoptosis-associated speck-like protein(ASC)in the cortex,hippocampus,and basal ganglia of rats following MCC was significantly increased,as shown by Western Blot analysis(P＜0.05),and cortical areas are more elevated.The expression of these proteins in different brain regions was reduced by CBD-10 and CBD-40 intervention(P＜0.05).Conclusion:Cannabidiol can reduce the inflammatory response of multiple cerebral concussions rats through NLRP3 inflammasome and protect nerve tissue.

Objective: To analyze the characteristics of influenza virus detection in an influenza outbreak in schools, so as to provide a strategic basis for the treatment of influenza outbreaks in schools. Methods: A total of 1 702 samples were collected from 52 school influenza outbreaks reported in Linyi City in 2021-2022. The samples were divided into 3 types according to different symptoms during the management of the epidemic [group A:influenzalike illness (ILI) group; group B:mild illness group; group C:close contacts group]. Rt-PCR was used to detect influenza virus nucleic acid in the collected samples. The detection rate of influenza virus in the outbreaks was analyzed by χ2 test. Results: In total, 1 071 samples (62.93%) tested positive for influenza virus nucleic acid. Among them, 610 out of 726 samples (84.02%) were detected in group A, while 331 out of 634 samples (52.21%) were detected in group B. In group C, 130 out of 342 samples (38.01%) tested positive. The differences were statistically significant (χ2=260.71, P<0.01). In group A, males had a detection rate of 80.83% for influenza virus nucleic acid, compared to 91.36% for females. For group B, the rates were 53.31% for males and 50.87% for females. In group C, males had a rate of 30.72%, while females had a rate of 43.92%. Statistical significance for gender differences was observed only in groups A and C (χ2=12.67, 6.25, P<0.05). According to the days of onset, the detection rates of influenza virus nucleic acid among patients with onset 0-6 days were 56.30%, 74.49%, 89.35%, 86.23%, 69.67%, 62.75%, 34.33%, respectively, and the difference was statistically significant (χ2=128.27, P<0.01). Conclusions Mild cases and close contacts are likely key factors contributing to the prolonged emergence of new cases within classrooms during school influenza outbreaks. The progression of influenza symptoms is related to the risk of transmission.

Objective:To investigate the effect of panax notoginseng saponins(PNS)on the expression of tumor necrosis factor-α(TNF-α)in lipopolysaccharide(LPS)-induced activated BV2 microglia through p38 mitogen-activa-ted protein kinase(p38 MAPK)pathway.Methods:BV2 microglia were divided into control group,LPS activated group and LPS+panax notoginseng saponins intervention group(LPS+PNS).The CCK-8 method was used to detect the viability of BV2 microglia and determine the optimal drug intervention concentration.Western Blot and immunofluo-rescence were used to detect the expression of p38 MAPK and TNF-α and the phosphorylation level of p38 MAPK(p-p38 MAPK)in BV2 microglia.Results:Compared with the blank control group,there was no significant difference in the cell viability of BV2 microglia,and finally 100 mg/L was selected as the drug intervention concentration.Western Blot and immunofluorescence results indicated that after LPS activation,the expression of TNF-α and the phosphoryla-tion level of p38 MAPK in BV2 microglia were significantly increased(P＜0.05).After PNS intervention,compared with LPS-activated group,the expression of TNF-α and the phosphorylation level of p38 MAPK were significantly decreased(P＜0.05).After treatment with p38 MAPK pathway inhibitor(SB203580),there was no significant differ-ence in the expression levels of p-p38 MAPK and TNF-α in PNS combined with SB203580 group(LPS+PNS+I)com-pared with LPS+PNS group(P＞0.05).In addition,the changes of p38 MAPK in each group were not statistically sig-nificant(P＞0.05).Conclusion:PNS may inhibit the expression of inflammatory factor TNF-α secreted by activated BV2 microglia through p38 MAPK pathway.

Objective:To explore the impact of scutellarin on the levels of phosphatidylinositol 3-kinase(P13K)and phosphorylated P13K(p-PI3K)in activated microglia.Methods:A rat cerebral ischemia model was made by middle cerebral artery occlusion(MCAO)method.SD rats were randomly divided into three groups:sham operation group(Sham),cerebral ischemia group(MCAO),and cerebral ischemia with scutellarin group(MCAO+S).A model of ischemia-hypoxia injury of BV2 microglia was established by glucose-oxygen deprivation(OGD)and randomly assigned as control group(Control),OGD group and OGD+scutellarin group(OGD+S).Changes in PI3K and p-PI3K expres-sion in microglia were assessed using double immunofluorescence staining and Western Blot.Results:Immunofluores-cence staining and Western Blot analyses revealed a significant increase in p-PI3K levels in activated microglia both in vivo and in vitro(P＜0.05).Furthermore,treatment with scutellarin led to a further elevation in p-PI3K(P＜0.05).However,there were no significant alterations observed in PI3 K expression among the groups(P＞0.05).Conclusion:Scutellarin may play a positive role in the treatment of cerebral ischemia by up-regulating the phosphorylation level of P13K in activated microglia.