1.Association of parenting style with emotional and behavioral problems in preschool children
LIANG Xi, LI Ruoyu, FANG Jiao, WANG Xiaoyan, WANG Yuhui
Chinese Journal of School Health 2025;46(9):1266-1271
Objective:
To explore the effects of parenting style on emotional and behavioral problems (EBPs) of preschool children and to provide theoretical reference for promoting children s physical and mental health.
Methods:
In October-November 2021, a stratified cluster random sampling method was adopted to select 3 575 preschool children aged 3-6 years from 12 kindergartens in Hefei. Data on parenting styles and children s EBPs were collected through electronic questionnaires. Binary Logistic regression model was used to explore the effects of parenting styles on children s EBPs.
Results:
The detection rates of emotional symptoms, conduct problems, hyperactivity, peer interaction problems, and prosocial behavior problems were 15.5% ( n =554), 23.2% ( n =830), 22.4% ( n =802), 38.2% ( n =1 367), and 29.5% ( n =1 055) in preschool children, respectively. Binary Logistic regression analysis showed that after adjusting for covariates such as child s age, sex, and preterm birth, a high authoritative parenting style reduced the risk of EBPs in children ( OR =0.29-0.78), while a high authoritarian ( OR =1.36-2.15) and high permissive ( OR =1.36-1.68) parenting style in mothers increased the risk of EBPs in children (all P <0.05). Further stratified analysis indicated that among children with fathers exhibiting high authoritative, high authoritarian, or high permissive parenting styles, a high authoritative parenting style in mothers reduced the risk of peer interaction problems and prosocial behavior problems ( OR =0.51, 0.30 ; 0.44, 0.23; 0.51, 0.29, all P <0.05). Among children with fathers exhibiting a high authoritative parenting style, a high authoritarian parenting style in mothers increased the risk of emotional problems ( OR =2.59), and both high authoritarian and high permissive parenting styles in mothers increased the risk of conduct problems ( OR =3.25, 2.09) and hyperactivity problems ( OR =2.35, 2.87) (all P <0.05). Among children with fathers exhibiting high authoritarian or high permissive parenting styles, a high authoritarian parenting style in mothers increased the risk of EBPs ( OR =1.65-2.71, 1.62-2.52, all P <0.05).
Conclusion
Parenting style is an important factor affecting EBPs of preschool children, and appropriate parenting style is beneficial for children s psychological development.
2.Developmental toxicity of Cry1Ab protein in the embryonic stem-cell model
Yuanzhi JIAN ; Fei WANG ; Ning YIN ; Ruoyu ZHOU ; Junbo WANG
Journal of Peking University(Health Sciences) 2024;56(2):213-222
Objective:To evaluate the developmental toxicity of Cry1Ab protein by studying its effects on cell proliferation and differentiation ability using a developmental toxicity assessment model based on embryonic stem-cell.Methods:Cry1Ab protein was tested in seven dose groups(31.25,62.50,125.00,250.00,320.00,1 000.00,and 2 000.00 μg/L)on mouse embryonic stem cells D3(ES-D3)and 3T3 mouse fibroblast cells,with 5-fluorouracil(5-FU)used as the positive control and phos-phate buffer saline(PBS)as the solvent control.Cell viability was detected by CCK-8 assay to calculate the 50%inhibitory concentration(IC50)of the test substance for different cells.Additionally,Cry1 Ab protein was tested in five dose groups(125.00,250.00,320.00,1 000.00,and 2 000.00 μg/L)on ES-D3 cells,with PBS as the solvent control and 5-FU used for model validation.After cell treatment,cardiac differentiation was induced using the embryonic bodies(EBs)culture method.The growth of EBs was observed under a microscope,and their diameters on the third and fifth days were measured.The proportion of EBs differentiating into beating cardiomyocytes was recorded,and the 50%inhibition con-centration of differentiation(ID50)was calculated.Based on a developmental toxicity discrimination func-tion,the developmental toxicity of the test substances was classified.Furthermore,at the end of the cul-ture period,mRNA expression levels of cardiac differentiation-related markers(Oct3/4,GATAA-4,Nkx2.5,and β-MHC)were quantitatively detected using real-time quantitative polymerase chain reaction(qPCR)in the collected EBs samples.Results:The IC50 of 5-FU was determined as 46.37 μg/L in 3T3 cells and 32.67 μg/L in ES-D3 cells,while the ID50 in ES-D3 cells was 21.28 μg/L.According to the discrimination function results,5-FU was classified as a strong embryotoxic substance.There were no sta-tistically significant differences in cell viability between different concentrations of Cry 1 Ab protein treat-ment groups and the control group in both 3T3 cells and ES-D3 cells(P>0.05).Moreover,there were no statistically significant differences in the diameter of EBs on the third and fifth days,as well as their morphology,between the Cry1Ab protein treatment groups and the control group(P>0.05).The cardi-ac differentiation rate showed no statistically significant differences between different concentrations of Cry1Ab protein treatment groups and the control group(P>0.05).5-FU significantly reduced the mRNA expression levels of β-MHC,Nkx2.5,and GATA-4(P<0.05),showing a dose-dependent trend(P<0.05),while the mRNA expression levels of the pluripotency-associated marker Oct3/4 exhibited an increasing trend(P<0.05).However,there were no statistically significant differences in the mRNA expression levels of mature cardiac marker β-MHC,early cardiac differentiation marker Nkx2.5 and GATA-4,and pluripotency-associated marker Oct3/4 between the Cry1Ab protein treatment groups and the control group(P>0.05).Conclusion:No developmental toxicity of Cry1Ab protein at concen-trations ranging from 31.25 to 2 000.00 μg/L was observed in this experimental model.
3.Application and prospect of robotic-assisted total knee arthroplasty
Ruoyu WANG ; Zhifeng ZHANG ; Jian HUANG
Chinese Journal of Tissue Engineering Research 2024;28(30):4889-4895
BACKGROUND:The basic principle of the design of the robot system used for total knee arthroplasty is to combine three-dimensional surgical planning,early warning of danger areas during surgery,real-time data feedback,robot arm assisted osteotomy and other technologies to achieve precision and personalization of total knee arthroplasty,which is exactly its biggest advantage.Therefore,it has become a hot topic in the field of joint surgery in recent years and attracted much attention. OBJECTIVE:To summarize the development status of robotic-assisted total knee arthroplasty in the field of joint surgery and the comparison of advantages and disadvantages with conventional total knee arthroplasty and to prospect the future development of robotic-assisted total knee arthroplasty. METHODS:Relevant articles were searched from PubMed,CNKI,Wanfang and VIP databases by computer.English key words were"robot OR robotic OR robotics OR robotically OR computer,total knee arthroplasty OR total knee replacement,TKA OR TKR".Chinese key words were"robotic-assisted,computer navigation,total knee arthroplasty".Finally,64 articles were included for review and analysis. RESULTS AND CONCLUSION:(1)The robot system used to assist total knee arthroplasty is divided into active,semi-active and passive according to its degree of freedom.The semi-active system,currently widely used in robotic systems,effectively enhances the accuracy and personalization of total knee arthroplasty.However,its high implementation cost and relatively steep learning curve remain key factors to be balanced when promoting its adoption in joint surgery field.(2)Robotic-assisted total knee arthroplasty can achieve precise osteotomy and correct placement of prosthesis in local three-dimensional space of the knee joint.It has been widely proven that it can provide better accuracy of prosthesis implantation,reduce imaging abnormalities,obtain good soft tissue balance during the operation,and ultimately improve the motion and functional status of the knee joint after the operation.(3)However,the current robotic-assisted system still has objective shortcomings,including the problem of learning curve between different robot devices and operators,additional installation and maintenance costs,and potential complications related to robot surgery.Therefore,whether it can truly benefit the medical system and patients still needs to be proven by long-term research,and the robotic-assisted system also needs to be further improved substantially.(4)Robotic-assisted total knee arthroplasty technique is still in the preliminary research stage in clinical practice and has not been widely applied.To better define the usage of robotic-assisted total knee arthroplasty and enhance its clinical procedural standards and safety,refining these aspects will become a focal point of future research on robotic-assisted total knee arthroplasty.
4.Patient-derived parotid gland pleomorphic adenoma organoid model and its preliminary histological and functional characterization
Yuxin CHEN ; Ruoyu WANG ; Qiuxu WANG ; Ruiqing LIAN ; Yu'e WANG ; Shanshan LIANG
Chinese Journal of Stomatology 2024;59(6):604-609
Objective:To establish patient-derived organoid models of pleomorphic adenomas (PA) of the parotid gland and preliminarily characterize their histology, related biomarkers and functions.Methods:Fresh tumor tissue specimens were collected from surgical procedures of Oral and Maxillofacial Department. The harvested tissues were processed and cultured in a head and neck tumor organoid culture system to establish organoid models from parotid gland pleomorphic adenomas. The in vitro growth of PA organoids was recorded by light microscopy. The successfully established organoids were passaged and cryopreserved, and the cryopreserved PA organoids were revived and re-cultured to observe their viability and organoid regeneration ability. Histological characterization, as well as characterization and detection of related markers and functional proteins, were performed on the organoids, comparing them with the patient-derived tissues. Results:The constructed organoid model of pleomorphic adenoma exhibited a dense and compact three-dimensional spherical structure. Hematoxylin and eosin staining indicated morphological similarities between the organoid and its tissue of origin. Immunohistochemistry showed positive cytoplasmic staining for Calponin, cytokeratin 7, and epithelial membrane antigen in both the organoid and the source tumor tissue, suggesting consistent histopathological characteristics between the organoid and its tissue of origin. Periodic acid-Schiff staining of the organoid showed positive staining for glycogen, with positive staining located in the interior and periphery of the organoid, indicating that the organoid possessed secretory functions like the salivary gland.Conclusions:This study successfully constructed organoids of pleomorphic adenoma derived from patient samples. This model faithfully replicates the tissue morphology and biomarkers of the source tissue and exhibits biological functions associated with mucus secretion. It serves as a valuable in vitro model for studying the development and progression of salivary gland tumors.
5.Comparison of dermoscopic features of toenail psoriasis and fingernail psoriasis
Shiqi WANG ; Aiping WANG ; Hang LI ; Ruoyu LI
Chinese Journal of Dermatology 2024;57(2):161-165
Objective:To evaluate and compare dermoscopic features of toenail psoriasis and fingernail psoriasis.Methods:Between June 2020 and January 2022, 61 patients with confirmed toenail psoriasis and 80 with confirmed fingernail psoriasis were enrolled from the Department of Dermatology of Peking University First Hospital. Dermoscopy was performed on 139 affected toenails and 158 affected fingernails among the psoriasis patients, and dermoscopic characteristics were analyzed between the two groups by using the chi-square test.Results:The most common dermoscopic feature of nail psoriasis was pitting (223/297, 75.08%), followed by splinter haemorrhages (164/297, 55.22%), subungual hyperkeratosis (133/297, 44.78%), oil drop sign (126/297, 42.42%), complete onycholysis (121/297, 40.74%), linear margin of the proximal onycholysis (107/297, 36.03%) and linear erythema at the margin of the onycholysis (77/297, 25.93%). Compared with the patients with fingernail psoriasis, those with toenail psoriasis more commonly presented with subungual hyperkeratosis (81[58.27%] vs. 52[32.91%], P < 0.001), punctate/blocky haemorrhages (22[15.83%] vs. 11[6.96%], P < 0.05), longitudinal striae (34[24.46%] vs. 10[6.33%], P < 0.001), longitudinal nail splitting (24[17.27%] vs. 9[5.70%], P < 0.01), brown discoloration (14[10.07%] vs. 2[1.27%], P < 0.01), transverse grooves (17[12.23%] vs. 1[0.63%], P < 0.001) and leukonychia (10[7.19%] vs. 1[0.63%], P < 0.01) ; compared with the patients with toenail psoriasis, those with fingernail psoriasis more commonly presented with splinter haemorrhages (100[63.29%] vs. 64[46.04%], P < 0.01), oil drop sign (81[51.27%] vs. 45[32.37%], P < 0.01), linear erythema at the margin of the onycholysis (55[34.81%] vs. 22[15.83%], P < 0.001), partial onycholysis (50[31.65%] vs. 19[13.67%], P < 0.001) and red spots in the lunula (36[22.78%] vs. 12[8.63%], P < 0.01) . Conclusion:The dermoscopic features of toenail psoriasis were quite different from those of fingernail psoriasis, and features such as subungual hyperkeratosis, longitudinal streaks, and brown discoloration were more commonly presented in patients with toenail psoriasis.
6.Experimental Study on Regulation of Nrf2/HO-1 by Linalool to Inhibit Hepatic Injury Induced by Aflatoxin B1
Meng WANG ; Chunmiao XUE ; Xin HUANG ; Wenhui LIU ; Ruoyu GAO ; Xuehui BAI ; Guodong HUA ; Baochen ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):89-96
ObjectiveTo investigate the effect of linalool against acute liver injury induced by aflatoxin B1(AFB1) in rats and explore its protective mechanism. MethodTwenty male SPF SD rats were randomly divided into three groups: Control (n=6), AFB1 (n=7), and linalool (n=7) groups. Linalool solution (200 mg·kg-1) was administered preventatively for 14 days, while the control and AFB1 groups intragastrically received an equivalent volume of double distilled water. After preventative administration of linalool, AFB1 solution (1 mg·kg-1, dissolved in saline) was intraperitoneally injected for two consecutive days to induce acute liver injury in rats. Samples were collected and processed 14 days after model establishment. Pathological changes in liver tissue of rats were observed using Hematoxylin-eosin(HE) staining and Masson staining. Biochemical detection was performed to measure the levels of alanine transaminase(ALT), aspartate transaminase(AST), γ-glutamyl transferase(GGT), lactate dehydrogenase(LDH), alkaline phosphatase(ALP), total bilirubin(TBil), direct bilirubin(DBil), indirect bilirubin(IBil), malondialdehyde(MDA), superoxidedismutase(SOD), catalase(CAT) , glutathione(GSH), Fe3+, and Fe2+ in the liver tissue. Western blot was adopted to assess protein expression levels of nuclear factor-erythroid 2-related factor 2(Nrf2) and heme oxygenase-1(HO-1). Molecular docking was performed to verify the binding between linalool and key proteins of the Nrf2/HO-1 signaling pathway. Molecular dynamics techniques were used to confirm the stability and affinity of linalool binding with key proteins of the Nrf2/HO-1 signaling pathway. ResultPathological results showed that compared to that in the AFB1 group, the liver structure in the linalool group tended to be normal, with a significant decrease in blue collagen fibers. The linalool group exhibited significantly reduced levels of ALT, AST, GGT, LDH, ALP, TBil, DBil, and IBil (P<0.01), Fe3+ and Fe2+ content, and oxidative stress marker MDA (P<0.01). The levels of antioxidants SOD, CAT, and GSH significantly increased (P<0.01). Molecular docking showed a molecular docking energy between linalool and Nrf2 and HO-1 targets of -5.495 6 and -5.199 4 kcal·mol-1(1 cal≈4.186 J), respectively. Molecular dynamics results indicated strong affinity in the binding of linalool with Nrf2 and HO-1. Western blot revealed a significant increase in Nrf2 protein expression (P<0.05) and a decrease in HO-1 protein expression (P<0.01) in the linalool group. ConclusionLinalool may protect against AFB1-induced acute liver injury by modulating the Nrf2/HO-1 ferroptosis signaling pathway to inhibit liver cell ferroptosis and regulate hepatic oxidative stress levels.
7.Investigation on Preventive Effect of Total Saponins of Notoginseng Radix et Rhizoma on Aspirin-induced Small Intestine Injury Based on Serum Metabolomics
Wenhui LIU ; Guodong HUA ; Baochen ZHU ; Ruoyu GAO ; Xin HUANG ; Meng WANG ; Zheng LIU ; Jiaojiao CHENG ; Zhibin SONG ; Jingui WANG ; Chunmiao XUE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(22):196-203
ObjectiveMetabolomics was utilized to investigate the preventive effect of notoginseng total saponins(NTS) on aspirin(acetyl salicylic acid, ASA)-induced small bowel injury in rats. MethodFifty male SD rats were randomly divided into normal and model groups, NTS high-dose and low-dose groups(62.5, 31.25 mg·kg-1), and positive drug group(omeprazole 2.08 mg·kg-1+rebamipide 31.25 mg·kg-1), with 10 rats in each group. Except for the normal group, rats in other groups were given ASA enteric-coated pellets 10.41 mg·kg-1 daily to establish a small intestine injury model. On this basis, each medication group was gavaged daily with the corresponding dose of drug, and the normal group and the model group were gavaged with an equal amount of drinking water. Changes in body mass and fecal characteristics of rats were recorded and scored during the period. After 14 weeks of administration, small intestinal tissues of each group were taken for hematoxylin-eosin(HE) staining, scanning electron microscopy to observe the damage, and the apparent damage of small intestine was scored. Serum from rats in the normal group, the model group, and the NTS high-dose group was taken and analyzed for metabolomics by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS), and the data were processed by multivariate statistical analysis, the potential biomarkers were screened by variable importance in the projection(VIP) value≥1.0, fold change(FC)≥1.5 or ≤0.6 and t-test P<0.05, and pathway enrichment analysis of differential metabolites was performed in conjunction with Human Metabolome Database(HMDB) and Kyoto Encyclopedia of Genes and Genomes(KEGG). ResultAfter 14 weeks of administration, the average body mass gain of the model group was lower than that of the normal group, and the NTS high-dose group was close to that of the normal group. Compared with the normal group, the fecal character score of rats in the model group was significantly increased(P<0.05), and compared with the model group, the scores of the positive drug group and the NTS high-dose group were reduced, but the difference was not statistically significant. HE staining and scanning electron microscopy results showed that NTS could significantly improve ASA-induced small intestinal injury, compared with the normal group, the small bowel injury score of the model group was significantly increased(P<0.01), compared with the model group, the small bowel injury scores of the NTS low and high dose groups were significantly reduced(P<0.05, P<0.01). Serum metabolomics screened a total of 75 differential metabolites between the normal group and the model group, of which 55 were up-regulated and 20 were down-regulated, 76 differential metabolites between the model group and the NTS groups, of which 14 were up-regulated and 62 were down-regulated. NTS could modulate three differential metabolites(salicylic acid, 3-hydroxybenzoic acid and 4-hydroxybenzoic acid), which were involved in 3 metabolic pathways, namely, the bile secretion, the biosynthesis of folic acid, and the biosynthesis of phenylalanine, tyrosine and tryptophan. ConclusionNTS can prevent ASA-induced small bowel injury, and the underlying mechanism may be related to the regulation of bile secretion and amino acid metabolic pathways in rats.
8.Effective Components and Mechanism of Cistanches Herba Aqueous Extract Against Diabetic Nephropathy Based on the Spectral-effect Relationship
Tianhua LIU ; Chong MA ; Baicai WANG ; Ruoyu GENG ; Yao ZHAO ; Junping HU ; Jianhua YANG
Chinese Journal of Modern Applied Pharmacy 2024;41(10):1313-1323
OBJECTIVE
To screen out the pharmacodynamic substances of Cistanches Herba aqueous extracts, explore the basis of the pharmacodynamic substances and their mechanism of action in the treatment of diabetic nephropathy(DN), based on the spectral relationship between the mass spectral peak areas of different elution sites of Cistanches Herba aqueous extracts and their anti-DN effects.
METHODS
UPLC-Orbitrap-MS/MS technique was used to characterise the chromatographic peaks; MTT method was used to detect the effects of different elution sites of Cistanches Herba aqueous extract on the proliferation of high glucose and high fat HK-2 cells; grey correlation analysis and partial least squares method were used to analyse the spectral relationship between mass spectrometry peak area and anti-DN activity. MTT method was used to determine the anti-DN activities of the individual components of Cistanches Herba aqueous extract; biochemical kit and ELISA were used to determine the levels of oxidative indicators(SOD, GSH-P) and inflammatory factors(IL-1β, TNF-α, TGF-β); Western blotting was used to detect the expression of apoptosis-related proteins.
RESULTS
UPLC-Orbitrap-MS/MS technique speculated and identified 72 common compounds; In Cistanches Herba aqueous extracts, water, 20% ethanol, and 40% ethanol-eluted sites differentially increased the proliferation rate of HK-2 cells in a high-sugar, high-fat environment; Partial least squares and grey correlation analyses showed that the constituents of the aqueous extracts of Cistanches Herba with greater anti-DN contributions were 8-epideoxymatricinic acid, geniposidic acid, pinoresinol, betaine and syringin, et al. MTT assay reveals that 8-epi-deoxystrychnic acid and geniposidic acid had significant proliferative effects on HK-2 cells in a high glucose and high fat environment; Biochemical kit and ELISA showed that 8-epideoxystrychnic acid and geniposidic acid were able to up-regulate the activity of SOD and GSH-Px, and at the same time they had an inhibitory effect on the expression of inflammatory factors IL-1β, TNF-α and TGF-β. Western blotting results showed that 8-epideoxystrychnic acid and geniposidic acid were able to down-regulate and up-regulate the markers of dermal mesenchymal transition: α-SMA, Collagen Ⅰ and E-cadherin, and they could exert anti-DN effects by inhibiting the PI3K-Akt pathway.
CONCLUSION
The two compounds, 8-epideoxystrychnic acid and geniposidic acid, which are screened by the spectroeffective relationship of anti-DN among the many chemical constituents contained in Cistanches Herba, can affect oxidative stress, inflammation and epithelial-mesenchymal transformation of renal tubular cells by inhibiting the PI3K-Akt signalling pathway, and improve renal pathology, degree of fibrosis and renal function, which will be useful for the in-depth study of aqueous extracts of Cistanches Herba in the treatment of DN.
9.Effect and mechanism of ammonium pyrrolidine dithiocarbamate on neuroinflammation in rats with traumatic brain injury
Jiafa YANG ; Zhaofeng LU ; Yaqiong WANG ; Ruoyu LU ; Hairong LI ; Mengjia LIU
Journal of Xinxiang Medical College 2024;41(7):631-639
Objective To investigate the effect and mechanism of ammonium pyrrolidine dithiocarbamate(PDTC)on neuroinflammatory injury in the penumbra of traumatic brain injury(TBI)in rats.Methods Sixty Sprague Dawley rats were divided into the PDTC group,TBI group,sham operation group and control group according to the random number table method,with 15 rats in each group.Rats in the PDTC group were intraperitoneally injected with PDTC(100 mg·kg-1)at 15 minutes before surgery;while the rats in the TBI group,sham operation group,and control group were intraperitoneally injected with the same volume of double distilled water.After the cranial window of rats in the TBI group and PDTC group was created,a 2.5 g steel rod with an inner diameter of 6.0 mm was dropped freely from a height of 75 cm through a transparent polyvinyl chloride tube with an inner diameter of 7.0 mm to impact the dura mater and induce right parietal lobe contusion and laceration to establish the TBI model;rats in the sham operation group were sealed with bone wax after the cranial window creation,without any impact applied;rats in the control group were raised under normal conditions.The modified neurological severity score(mNSS)was used to evaluate the degree of neurobehavioral damage in rats in each group at 1,4 and 7 days after modeling.At 2 days after modeling,5 rats in each group were decapitated,and brain tissues were taken for hematoxylin & eosin(HE)staining,and morphological changes of the brain tissues were observed under an optical microscope.The expressions of β-amyloid precursor protein(β-APP)and glial fibrillary acidic protein(GFAP)in the brain tissues of rats in each group were detected by immunohistochemical staining.At 24 hours after modeling,5 rats in each group were decapitated,and the right injured penumbra tissues were obtained;the expressions of nuclear transcription factor-κB(NF-κB)P65,phosphorylated NF-κB P65,inhibitor of NF-κB(IκB),phosphorylated IKB,NOD-like receptor protein 3(NLRP3)and caspase-1 protein in the right injured penumbra tissue of rats in each group were detected by Western blot,and the expressions of NF-κB P65,IκB,NLRP3 and caspase-1 mRNA in the right injured penumbra tissue of rats in each group were determined by real-time quantitative polymerase chain reaction.Results At 1,4,and 7 days after modeling,the mNSS scores of rats in the TBI group were signifi-cantly higher than those in the PDTC group,control group and sham operation group.The mNSS scores of rats in the PDTC group were significantly higher than those in the control group and sham operation group(P<0.05);there was no statistically significant difference in mNSS scores between the sham operation group and the control group(P>0.05).The neurons and neurogliocyte of rats in the control group and the sham operation group exhibited normal morphology,without swelling and wide-ning of intercellular space.Diffuse hemorrhagic changes were observed in the brain tissues of rats in the TBI group,with different morphologies of neuronal cell body,unclear cell membrane and cytoplasm,pyknosis of cell nuclei,often triangular shape,disappearance of normal structure and nucleoli,and diffuse white blood cells and red blood cells filling the field of vision.The lesion surrounding area of rats in the PDTC group showed ischemic changes,with mild shrinkage of neuronal volume,a uniform light red color,karyopyknosis,nuclear-cytoplasmic dissociation,disappearance of normal structure and nucleoli,and localization of neuroinflammation.There was no significant expression of β-APP and GFAP in the cerebral cortex of rats in the control group and the sham operation group,while the accumulation of β-APP and GFAP in neuronal serosae and/or axons was observed in the brain tissues of rats in the TBI group and the PDTC group.Compared with the TBI group,a decrease in the number and the expression intensity of β-APP and GFAP-positive stained neuronal cells in the cerebral cortex of rats was observed in the PDTC group.The relative expression of NF-κB P65 protein in the brain tissues of rats in the sham operation group,TBI group and PDTC group was significantly higher than that in the control group,and the relative expression of NF-κB P65 protein in the brain tissues of rats in the PDTC group was significantly higher than that in the TBI group(P<0.05).The relative expression of phosphorylated NF-κB P65 protein in the brain tissues of rats in the PDTC group and the sham operation group was significantly lower than that in the control group,the relative expression of phosphorylated NF-κB P65 protein in the brain tissues of rats in the TBI group was significantly higher than that in the control group and the sham operation group,and the relative expression of phosphorylated NF-κB P65 protein in the brain tissues of rats in the PDTC group was significantly lower than that in both TBI group and sham operation group(P<0.05).There was no significant difference in the relative expression of IκB protein in the brain tissues of rats between the sham operation group and the control group(P>0.05);the relative expression of IκB protein in the brain tissues of rats in the TBI group was significantly higher than that in the control group,and the relative expression of IκB protein in the brain tissues of rats in the PDTC group was significantly lower than that in the control group,sham operation group,and TBI group(P<0.05).The relative expression of phosphorylated IκB protein in the brain tissues of rats in the TBI group was significantly lower than that in the control group and the sham operation group,and the relative expression of phosphorylated IκB protein in the brain tissues of rats in the PDTC group was significantly higher than that in the TBI group(P<0.05).The relative expression of NLRP3 protein in the brain tissues of rats in the sham opera-tion group was significantly higher than that in the control group,the relative expression of NLRP3 protein in the brain tissues of rats in the TBI group and the PDTC group was significantly lower than that in the sham operation group and the control group,and the relative expression of NLRP3 protein in the brain tissues of rats in the TBI group was significantly lower than that in the PDTC group(P<0.05).The relative expression of caspase-1 protein in the brain tissues of rats in the sham opera-tion group,PDTC group,and TBI group was significantly higher than that in the control group,and the relative expression of caspase-1 protein in the brain tissues of rats in the PDTC group was significantly lower than that in the TBI group(P<0.05).The relative expression of NF-κB P65 mRNA in the brain tissues of rats in the PDTC group,TBI group,and sham operation group was significantly higher than that in the control group,the relative expression of NF-κB P65 mRNA in the brain tissues of rats in the PDTC group and TBI group was significantly higher than that in the sham operation group,and the relative expres-sion of NF-κB P65 mRNA in the brain tissues of rats in the PDTC group was significantly lower than that in the TBI group(P<0.05).The relative expression of IκB mRNA in the brain tissues of rats in the PDTC group and TBI group were signifi-cantly higher than that in the control group,and the expression of IκB mRNA in the brain tissues of rats in the sham operation group was significantly lower than that in the control group(P<0.05).The relative expression of IκB mRNA in the brain tis-sues of rats in the PDTC group and TBI group was significantly higher than that in the sham operation group,and the relative expression of IκB mRNA in the brain tissues of rats in the PDTC group was significantly lower than that in the TBI group(P<0.05).The relative expression of NLRP3 mRNA in the brain tissues of rats in the PDTC group and TBI group was significantly higher than that in the control group,the relative expression of NLRP3 mRNA in the brain tissues of rats in the sham operation group was significantly lower than that in the control group,the relative expression of NLRP3 mRNA in the brain tissues of rats in the PDTC group and TBI group was significantly higher than that in the sham operation group,and the relative expression of NLRP3 mRNA in the brain tissues of rats in the PDTC group was significantly lower than that in the TBI group(P<0.05).The relative expression of caspase-1 mRNA in the brain tissues of rats in the sham operation group,TBI group,and PDTC group was significantly lower than that in the control group,the relative expression of caspase-1 mRNA in the brain tissues of rats in the TBI group and PDTC group was significantly higher than that in the sham operation group(P<0.05).Conclusion PDTC can effectively improve neural functional deficit score and reduce neuroinflammatory injury in TBI rats,the mechanism of which may be related to regulating mRNA and protein expression of NF-κB/NLRP3 axis-related inflammatory injury indicators and regulating downstream inflammatory factors.
10.Isopimpinelline mitigates the oxidative stress and inflammation triggered by pineal gland damage through inhibiting NF-κB signaling pathway
Qiming ZHONG ; Liguo TONG ; Huajie ZHAO ; Ruoyu WANG ; Meiqing SONG ; Chongzhi BAI ; Haijie JI ; Mali FENG
Immunological Journal 2024;40(3):234-241
This study was designed to investigate the potential protective effect of isopimpinelline against para-chlorophenylalanine(PCPA)-induced pineal gland damage in rats.Forty male Sprague-Dawley(SD)rats were divided into four groups(n=10 each):a normal group,a model group,a melatonin-treated group(10 mg/kg),and an isopimpinelline-treated group(1.5 mg/kg).All groups,except for the normal,received intraperitoneal injection of PCPA(450 mg/kg)to induce pineal gland damage.Subsequent treatments were administered orally for 7 days.Sleep latency and duration were evaluated on the sixth day using the pentobarbital sodium sleep synergy test.After the treatment period,serum melatonin levels and pineal gland inflammation markers were assessed alongside oxidative and antioxidative parameters.Histological examinations of the pineal gland were conducted,and the expression of proteins related to the Nrf2 and NF-κB signaling pathways were quantified.Data showed that isopimpinelline alleviates the structural damage in the pineal gland of model rats,significantly elevated serum melatonin levels,and markedly improved sleep latency and duration(P<0.05).Isopimpinelline activated the Nrf2 signaling pathway by inhibiting Keap1 expression,which facilitated the nuclear translocation of Nrf2 and upregulated the antioxidant proteins NQO1 and HO-1,thereby mitigating oxidative stress in the pineal gland(P<0.05).Furthermore,isopimpinelline significantly reduced the levels of pro-inflammatory cytokines IL-2,TNF-α and IL-6.Isopimpinelline also suppressed the NF-κB signaling pathway,reducing the expression of NF-κB p65,IKKβ,and p-IKKβ proteins,as well as the nuclear translocation of NF-κB p65(P<0.05),thereby providing anti-inflammatory benefits.In conclusion,isopimpinelline could protect pineal gland from damage by activating the Nrf2 signaling pathway and inhibiting the NF-κB pathway.


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