1.Expert consensus on the application of nasal cavity filling substances in nasal surgery patients(2025, Shanghai).
Keqing ZHAO ; Shaoqing YU ; Hongquan WEI ; Chenjie YU ; Guangke WANG ; Shijie QIU ; Yanjun WANG ; Hongtao ZHEN ; Yucheng YANG ; Yurong GU ; Tao GUO ; Feng LIU ; Meiping LU ; Bin SUN ; Yanli YANG ; Yuzhu WAN ; Cuida MENG ; Yanan SUN ; Yi ZHAO ; Qun LI ; An LI ; Luo BA ; Linli TIAN ; Guodong YU ; Xin FENG ; Wen LIU ; Yongtuan LI ; Jian WU ; De HUAI ; Dongsheng GU ; Hanqiang LU ; Xinyi SHI ; Huiping YE ; Yan JIANG ; Weitian ZHANG ; Yu XU ; Zhenxiao HUANG ; Huabin LI
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(4):285-291
This consensus will introduce the characteristics of fillers used in the surgical cavities of domestic nasal surgery patients based on relevant literature and expert opinions. It will also provide recommendations for the selection of cavity fillers for different nasal diseases, with chronic sinusitis as a representative example.
Humans
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Nasal Cavity/surgery*
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Nasal Surgical Procedures
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China
;
Consensus
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Sinusitis/surgery*
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Dermal Fillers
2.Pueraria lobata decoction intervenes in neuroinflammatory response and apoptosis in rats with cervical spondylotic myelopathy
Diyou WU ; Jiajun HUANG ; Guangyi TAO ; Yu ZHAO ; Junqing HUANG ; Bin YANG ; Yun XUE
Chinese Journal of Tissue Engineering Research 2025;29(20):4249-4257
BACKGROUND:Inflammation and apoptosis play key roles in the pathological process of cervical spondylotic myelopathy.Previous studies have shown that Pueraria lobata decoction has favorable therapeutic effects on cervical spondylotic myelopathy.OBJECTIVE:To investigate the effects and mechanism of Pueraria lobata decoction in neuroinflammatory response and apoptosis in rats with cervical spondylotic myelopathy.METHODS:Sixty Sprague-Dawley rats were randomly divided into six groups:a normal group,a sham-operated group,a model group,and three groups that received low,medium,and high doses of Pueraria lobata decoction.An animal model of cervical spondylotic myelopathy was constructed through compression of the spinal cord with water-absorbing and expanding material.Gastric administration of Pueraria lobata decoction(4.86,9.72,and 19.44 g/kg)was given in the three Pueraria lobata decoction groups 2 weeks after surgery,and the resting groups were given saline by gavage,once daily for 4 weeks.Motor function evaluation(Basso-Beattie-Bresnahan score)was performed in rats on days 1,7,14,21 and 28 after drug administration.At 4 weeks after drug administration,hematoxylin-eosin staining was used to observe the pathomorphologic changes in spinal cord tissue;immunofluorescence double staining was used for the detection of microglial cell polarization in spinal cord tissue;quantitative fluorescence PCR was used to detect the changes in the expression of interleukin-6 and interleukin-1β mRNA;western blot assay was used to detect the protein expression of p-NF-κB p65,NF-κB p65,NLRP3,ASC,Cleaved Caspase-1,Bax,Bcl-2,Cleaved Caspase-3,NOX4,p-Drp1,Drp1,and Mfn2 in spinal cord tissues;TUNEL assay was used to detect apoptosis in spinal cord tissues;and DHE staining was used to detect reactive oxygen species levels in rat spinal cord tissues.RESULTS AND CONCLUSION:(1)Compared with the normal and sham-operated groups,reduced Basso-Beattie-Bresnahan scores were observed in the model group(P<0.05),spinal cord neurons were crumpled and malformed with vacuolike changes.The Basso-Beattie-Bresnahan scores in the low-,medium-and high-dose Pueraria lobata decoction groups were significantly higher than those in the model group(P<0.05),and spinal cord neuronal damage reduced significantly.(2)Compared with the normal and sham-operated groups,there were elevated levels of Iba-1 and inducible nitric oxide synthase proteins and increased interleukin-6 and interluekin-1β mRNA expression in the spinal cord tissue of rats in the model group(P<0.05).The expression levels of Iba-1,inducible nitric oxide synthase,p-NF-κB,NLRP3,ASC and cleaved caspase-1 proteins as well as interleukin-6 and interleukin-1β mRNAs in the spinal cord tissues of rats in the low-,medium-and high-dose Pueraria lobata decoction groups were reduced compared with those in the model group(P<0.05).(3)Compared with the normal and sham-operated groups,the rate of TUNEL-positive cells and the levels of Bax and cleaved caspase-3 proteins were increased in the spinal cord tissues of rats in the model group(P<0.05),while the expression of Bcl-2 protein was reduced(P<0.05).Compared with the model group,the above indexes were significantly improved in the low-,medium-and high-dose Pueraria lobata decoction groups.(4)Compared with the normal and sham-operated groups,the model group exhibited increased levels of reactive oxygen species,along with elevated expression of NOX4 and p-Drp1 proteins(P<0.05)and reduced expression of Mfn2 protein(P<0.05)in the rat spinal crod tissue.Compared with the model group,the low-,medium-,and high-dose Pueraria lobata decoction groups exhibited reduced levels of reactive oxygen species,as well as decreased expression of NOX4 and p-Drp1 proteins(P<0.05)and increased expression of Mfn2 protein(P<0.05)in the rat spinal cord tissue.To conclude,Pueraria lobata decoction inhibits neuroinflammatory responses and neuronal apoptosis in the rat model of cervical spondylotic myelopathy,and the mechanism of action may be related to the regulation of the NOX4/reactive oxygen species/DRP1 signaling pathway.
3.Visualization analysis of research hotspots of artificial intelligence in field of spinal cord nerve injury and repair
Bin YANG ; Guangyi TAO ; Shun YANG ; Junjie XU ; Junqing HUANG
Chinese Journal of Tissue Engineering Research 2025;29(4):761-770
BACKGROUND:In recent years,artificial intelligence has gradually emerged and has been applied in various fields such as spinal cord nerve injury and repair,which has a positive impact on clinical treatment. OBJECTIVE:To study the application progress of artificial intelligence in the diagnosis,treatment,and rehabilitation of spinal cord nerve injury and repair,clarify the research hotspots and shortcomings in this field,and provide suggestions for future research work. METHODS:Relevant literature on artificial intelligence in the field of spinal cord nerve injury and repair was retrieved on the Web of Science core collection database until 2023.CiteSpace 6.1.R6 and VOSviewer 1.6.19 software was used to perform general literature analysis,co-citation of literature,co-citation of journals,double image overlay of journals,keyword clustering,and other visual analysis on the literature data. RESULTS AND CONCLUSION:(1)A total of 1 713 articles were selected,and the annual publication volume in this field showed a fluctuating upward trend,with the United States taking the lead,and Kadone and Hideki being the authors with the highest publication volume.ARCH PHYS MED REHAB was the journal with the highest number of citations.(2)Keyword co-occurrence and cluster analysis showed that after removing keywords similar to the search terms,the main keywords were divided into three main clusters:Exoskeleton and exercise rehabilitation(the largest core hotspot);machine learning and neural plasticity;robotics and rehabilitation training.(3)Keyword burst analysis showed that deep learning and artificial intelligence had become burst terms in the past five years.(4)The results of in-depth analysis of co cited and highly cited literature showed that the hotspots of artificial intelligence in the field of spinal cord nerve injury and repair were mainly focused on powered exoskeletons,gaits,electrical nerve stimulation,intracortical brain-computer interface(IBCI),robots,and polymer biomaterials,and neural stem cell.(5)The research on artificial intelligence in the field of spinal cord nerve injury and repair has shown an upward trend in recent years.The focus of this field had gradually shifted from single treatment methods such as exoskeletons and electrical stimulation to intelligent,precise,and personalized directions.(6)There were some limitations in this field,such as the consequences of missing or imbalanced data,low data accuracy and reproducibility,and ethical issues(such as privacy,research transparency,and clinical reliability).Future research should address the issue of data collection,requiring large sample,high-quality clinical datasets to establish effective artificial intelligence models.At the same time,the research on genomics and other mechanisms in this field is very weak.In the future,various machine learning technologies such as brain chips can be used,and gene editing therapy,single-cell spatial transcriptome and other methods can be used to study the basic mechanisms of regeneration-related gene upregulation and axon growth structural protein production.
4.Shentong Zhuyutang Regulates SIRT1/Nrf2 Pathway to Ameliorate Intervertebral Disc Degeneration in Rats
Jiajun HUANG ; Diyou WU ; Guangyi TAO ; Yu ZHAO ; Junqing HUANG ; Bin YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):29-39
ObjectiveTo study the effect and mechanism of Shentong Zhuyutang in treating intervertebral disc degeneration (IDD) in rats. MethodsIn the cell experiment, male rats were administrated with normal saline or low-, medium-, and high-dose (3.38, 6.75,13.5 g·kg-1, respectively) Shentong Zhuyutang by gavage, respectively, and serum samples were collected after 7 days of continuous administration. Another 10 male rats were selected for the isolation of nucleus pulposus cells. The cell model of IDD was established by treatment with interleukin (IL)-1β. The modeled cells were then treated with Shentong Zhuyutang-containing serum and the ferroptosis inhibitor ferrostatin-1 (Fer-1), respectively, to investigate the effects of Shentong Zhuyutang-containing serum on the proliferation and ferroptosis of nucleus pulposus cells. To study the role of silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) in the regulation of ferroptosis in nucleus pulposus cells by Shentong Zhuyutang-containing serum, this study treated the cells with the SIRT1 inhibitor Ex 527 and the Nrf2 inhibitor ML385, respectively, in addition to the treatment with IL-1β and high-dose Shentong Zhuyutang-containing serum. The cell-counting kit-8 (CCK-8) assay and EdU staining were employed to measure the cell viability and proliferation, respectively. The Fe2+, glutathione (GSH), and malondiadehyde (MDA) levels were measured by colorimetric assay. Western blot was employed to determine the protein levels of glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family 4 (ACSL4), Collagen Ⅱ, Aggrecan, SIRT1, and Nrf2. Immunofluorescence was used detect SIRT1 expression. In the animal experiment, male rats were treated with anulus puncture for the modeling of IDD. Rats were randomly assigned into sham operation, model, Shentong Zhuyutang-containing serum (13.5 g·kg-1), and positive control (nimesulide dispersible tablets, 0.18 mg·kg-1) groups. Rats in the drug intervention groups were administrated with corresponding agents at 1 mL·kg-1, and those in the sham operation and model groups were administrated with equal volumes of normal saline, once daily for 28 consecutive days. At the end of the last administration, the histopathological changes in the intervertebral discs of rats were observed by hematoxylin-eosin staining and scored by the Masuda method. Western blot was employed to determine the protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ in the nucleus pulposus tissue. ResultsCompared with the control group, the IL-1β group of nucleus pulposus cells showed elevated levels of Fe2+, MDA, and ACSL4 (P<0.05), decreased cell viability, lowered GSH level, and down-regulated protein levels of GPX4, Collagen Ⅱ, and Aggrecan (P<0.05). Shentong Zhuyutang-containing serum and Fer-1 reversed the effects of IL-1β on the viability and ferroptosis of nucleus pulposus cells and up-regulated the protein levels of Collagen Ⅱ and Aggrecan in nucleus pulposus cells (P<0.05). Compared with the control group, the IL-1β group showcased down-regulated expression of Sirt1 and Nrf2 in nucleus pulposus cells (P<0.05). Compared with the IL-1β group, the high-dose Shentong Zhuyutang-containing serum+IL-1β group showed up-regulated expression of SIRT1 and Nrf2 in nucleus pulposus cells (P<0.05). Compared with the high-dose Shentong Zhuyutang-containing serum+IL-1β group, the ML385 group showed down-regulated protein levels of Nrf2 and GPX4, lowered GSH level, and elevated Fe2+ and MDA levels (P<0.05). In addition, the Ex 527 group showed down-regulated protein levels of SIRT1, Nrf2, and GPX4 (P<0.05). The results of the animal experiment showed that compared with the sham operation group, the model group had severe degeneration of the intervertebral disc tissue with increased pathological score, up-regulated protein level of ACSL4 (P<0.05), and down-regulated protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ (P<0.05). Compared with the model group, the Shentong Zhuyutang group showed alleviated IDD with declined pathological score, down-regulated protein level of ACSL4 (P<0.05), and up-regulated protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ (P<0.05). ConclusionShentong Zhuyutang may activate the SIRT1/Nrf2 signaling pathway to inhibit the ferroptosis of nucleus pulposus cells, thereby delaying the process of IDD in rats.
5.Establishment and evaluation of a lipopolysaccharide-induced acute respiratory distress syndrome model in minipigs
Chuang-Ye WANG ; Ran WANG ; Jian ZHANG ; Ling-Xiao QIU ; Bin QING ; Heng YOU ; Jin-Cheng LIU ; Bin WANG ; Nan-Bo WANG ; Jia-Yu LI ; Xing LIU ; Shuang WANG ; Jin HU ; Jian WEN ; Quan LI ; Xiao-Ou HUANG ; Kun ZHAO ; Shuang-Lin LIU ; Gang LIU ; Mei-Ju WANG ; Qing XIANG ; Hong-Mei WU ; Xiao-Rong SUN ; Tao GU ; Dong ZHANG ; Qi LI ; Zhi XU
Medical Journal of Chinese People's Liberation Army 2025;50(9):1154-1161
Objective To establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.
6.Association of Body Mass Index with All-Cause Mortality and Cause-Specific Mortality in Rural China: 10-Year Follow-up of a Population-Based Multicenter Prospective Study.
Juan Juan HUANG ; Yuan Zhi DI ; Ling Yu SHEN ; Jian Guo LIANG ; Jiang DU ; Xue Fang CAO ; Wei Tao DUAN ; Ai Wei HE ; Jun LIANG ; Li Mei ZHU ; Zi Sen LIU ; Fang LIU ; Shu Min YANG ; Zu Hui XU ; Cheng CHEN ; Bin ZHANG ; Jiao Xia YAN ; Yan Chun LIANG ; Rong LIU ; Tao ZHU ; Hong Zhi LI ; Fei SHEN ; Bo Xuan FENG ; Yi Jun HE ; Zi Han LI ; Ya Qi ZHAO ; Tong Lei GUO ; Li Qiong BAI ; Wei LU ; Qi JIN ; Lei GAO ; He Nan XIN
Biomedical and Environmental Sciences 2025;38(10):1179-1193
OBJECTIVE:
This study aimed to explore the association between body mass index (BMI) and mortality based on the 10-year population-based multicenter prospective study.
METHODS:
A general population-based multicenter prospective study was conducted at four sites in rural China between 2013 and 2023. Multivariate Cox proportional hazards models and restricted cubic spline analyses were used to assess the association between BMI and mortality. Stratified analyses were performed based on the individual characteristics of the participants.
RESULTS:
Overall, 19,107 participants with a sum of 163,095 person-years were included and 1,910 participants died. The underweight (< 18.5 kg/m 2) presented an increase in all-cause mortality (adjusted hazards ratio [ aHR] = 2.00, 95% confidence interval [ CI]: 1.66-2.41), while overweight (≥ 24.0 to < 28.0 kg/m 2) and obesity (≥ 28.0 kg/m 2) presented a decrease with an aHR of 0.61 (95% CI: 0.52-0.73) and 0.51 (95% CI: 0.37-0.70), respectively. Overweight ( aHR = 0.76, 95% CI: 0.67-0.86) and mild obesity ( aHR = 0.72, 95% CI: 0.59-0.87) had a positive impact on mortality in people older than 60 years. All-cause mortality decreased rapidly until reaching a BMI of 25.7 kg/m 2 ( aHR = 0.95, 95% CI: 0.92-0.98) and increased slightly above that value, indicating a U-shaped association. The beneficial impact of being overweight on mortality was robust in most subgroups and sensitivity analyses.
CONCLUSION
This study provides additional evidence that overweight and mild obesity may be inversely related to the risk of death in individuals older than 60 years. Therefore, it is essential to consider age differences when formulating health and weight management strategies.
Humans
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Body Mass Index
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China/epidemiology*
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Male
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Female
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Middle Aged
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Prospective Studies
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Rural Population/statistics & numerical data*
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Aged
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Follow-Up Studies
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Adult
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Mortality
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Cause of Death
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Obesity/mortality*
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Overweight/mortality*
7.The effect of glutamine-enriched parenteral nutrition on short-term outcomes after surgery in patients with colorectal cancer
Ji-Wei WANG ; Yong HUANG ; Tao ZHANG ; Mai-Jian WANG ; Xin-Bin ZHENG ; Ming XIE
Parenteral & Enteral Nutrition 2025;32(1):7-14
Objective:To evaluate the effects of glutamine-enriched parenteral nutrition on short-term postoperative outcomes compared with conventional nutritional support in colorectal cancer patients with enteral nutrition intolerance after surgery.Method:A retrospective cohort study was conducted to collect clinical data from colorectal cancer patients who underwent radical resection at the Department of Gastrointestinal Surgery of Zunyi Medical University Affiliated Hospital from January 2019 to December 2021.The differences in postoperative complication rates,perioperative nutritional indicators,and inflammatory factors between the two groups were analyzed.SPSS 29.0 software was used for statistical analysis.Result:Based on whether glutamine was added to parenteral nutrition,178 patients were divided into a control group(conventional nutritional therapy,n=120)and an observation group(glutamine enhanced nutritional therapy,n=58).The incidence of postoperative complications(Clavien Dindo grade≥III)in the control and observation groups was 14.17%(17/120)and 3.45%(2/58),respectively,with a statistically significant difference(P=0.030).The observation group recovered faster than the control group in terms of time to first expectoration,defecation and intake of liquid diet after surgery,and had a shorter hospital stay after surgery(P<0.05).However,there was no significant difference in the 30-day readmission rate between the two groups(P=0.393).There was no statistically significant difference in the changes in total protein,albumin,pre-albumin,neutrophil/lymphocyte ratio and endogenous creatinine clearance rate between the two groups of patients after surgery(P>0.05).There were also significant differences in the changes in lymphocyte count,white blood cell count,neutrophil percentage,alanine aminotransferase,aspartate aminotransferase,total bilirubin,and urea nitrogen levels between the two groups of patients after surgery(P<0.05).Conclusion:Compared with regular nutritional support,postoperative parenteral glutamine supplementation can reduce the incidence of postoperative complications in colorectal cancer patients,promote recovery of bowel function,shorten postoperative hospital stay,improve patient immune function and reduce inflammatory levels.
8.Regulation of type Ⅰ interferon secretion via the RIG-Ⅰ signaling pathway after PRV infection of mouse trigeminal ganglion cells
Zhengbo LIAO ; Deyuan TANG ; Zhiyong ZENG ; Bin WANG ; Tao HUANG ; Xu CHEN ; Shen-glin YUAN ; Song HE ; Piao ZHOU ; Yinming MAO
Chinese Journal of Veterinary Science 2025;45(2):255-265
This study investigates the effects of pseudorabies virus(PRV)infection on the antiviral immune signaling pathways and type Ⅰ interferon factors in mouse trigeminal ganglion(TG)cells.In this experiment,primary TG cells were infected with PRV at a multiplicity of infection(MOI)of 1,while mice were infected via a drop-nose method using 106,29 TCID50 of PRV.Real-time fluorescence quantitative PCR(qPCR),Western blot and ELISA were used to assess gene tran-scription,protein expression,and the secretion of IFN-α and IFN-β.The results indicated that PRV infection of mouse TG primary cells led to alterations in the gene and protein expression of RIG-Ⅰ,MAVS,and IRF3,as well as the phosphorylation of IRF3 and IKBα both in vivo and ex vivo.ELISA results showed that PRV infection could regulate the secretion of IFN-α and IFN-β in mouse primary TG cells and mouse TGs.The results of RIG-Ⅰ signaling pathway-related proteins and the secretion of IFN-a and IFN-β were analyzed using Western blot after using siRNA to interfere with RIG-Ⅰ expression in TG cells.The results showed that siRIG-Ⅰ successfully inter-fered with RIG-Ⅰ protein expression in TG cells and caused changes in the expression of down-stream proteins such as MAVS and IRF3,and also regulated the secretion of IFN-α and IFN-β in TG cells.Furthermore,the results indicated that PRV infection induced the expression of RIG-Ⅰ in mouse TG progenitor cells,regulating the antiviral immune response of type Ⅰ interferon factors in TG cells through the RIG-Ⅰ-MAVS-IRF3 signaling axis.Notably,PRV inhibited the expression of IRF3 in TG cells while significantly upregulating the expression of IFN-β during the later stages of infection,which may be an important factor in the important reason for the rapid mortality ob-served in mice during the late stages of PRV infection.This experiment elucidates part of the anti-viral immune mechanism mediated by the RIG-Ⅰ-MAVS-IRF3 signaling pathway in regulating type Ⅰ interferon factor after PRV infection of mouse TG cells,as well as the discovery of differ-ent trends of IRF3 protein changes in vivo and ex vivo,laying the groundwork for future in-depth studies.
9.Pueraria lobata decoction intervenes in neuroinflammatory response and apoptosis in rats with cervical spondylotic myelopathy
Diyou WU ; Jiajun HUANG ; Guangyi TAO ; Yu ZHAO ; Junqing HUANG ; Bin YANG ; Yun XUE
Chinese Journal of Tissue Engineering Research 2025;29(20):4249-4257
BACKGROUND:Inflammation and apoptosis play key roles in the pathological process of cervical spondylotic myelopathy.Previous studies have shown that Pueraria lobata decoction has favorable therapeutic effects on cervical spondylotic myelopathy.OBJECTIVE:To investigate the effects and mechanism of Pueraria lobata decoction in neuroinflammatory response and apoptosis in rats with cervical spondylotic myelopathy.METHODS:Sixty Sprague-Dawley rats were randomly divided into six groups:a normal group,a sham-operated group,a model group,and three groups that received low,medium,and high doses of Pueraria lobata decoction.An animal model of cervical spondylotic myelopathy was constructed through compression of the spinal cord with water-absorbing and expanding material.Gastric administration of Pueraria lobata decoction(4.86,9.72,and 19.44 g/kg)was given in the three Pueraria lobata decoction groups 2 weeks after surgery,and the resting groups were given saline by gavage,once daily for 4 weeks.Motor function evaluation(Basso-Beattie-Bresnahan score)was performed in rats on days 1,7,14,21 and 28 after drug administration.At 4 weeks after drug administration,hematoxylin-eosin staining was used to observe the pathomorphologic changes in spinal cord tissue;immunofluorescence double staining was used for the detection of microglial cell polarization in spinal cord tissue;quantitative fluorescence PCR was used to detect the changes in the expression of interleukin-6 and interleukin-1β mRNA;western blot assay was used to detect the protein expression of p-NF-κB p65,NF-κB p65,NLRP3,ASC,Cleaved Caspase-1,Bax,Bcl-2,Cleaved Caspase-3,NOX4,p-Drp1,Drp1,and Mfn2 in spinal cord tissues;TUNEL assay was used to detect apoptosis in spinal cord tissues;and DHE staining was used to detect reactive oxygen species levels in rat spinal cord tissues.RESULTS AND CONCLUSION:(1)Compared with the normal and sham-operated groups,reduced Basso-Beattie-Bresnahan scores were observed in the model group(P<0.05),spinal cord neurons were crumpled and malformed with vacuolike changes.The Basso-Beattie-Bresnahan scores in the low-,medium-and high-dose Pueraria lobata decoction groups were significantly higher than those in the model group(P<0.05),and spinal cord neuronal damage reduced significantly.(2)Compared with the normal and sham-operated groups,there were elevated levels of Iba-1 and inducible nitric oxide synthase proteins and increased interleukin-6 and interluekin-1β mRNA expression in the spinal cord tissue of rats in the model group(P<0.05).The expression levels of Iba-1,inducible nitric oxide synthase,p-NF-κB,NLRP3,ASC and cleaved caspase-1 proteins as well as interleukin-6 and interleukin-1β mRNAs in the spinal cord tissues of rats in the low-,medium-and high-dose Pueraria lobata decoction groups were reduced compared with those in the model group(P<0.05).(3)Compared with the normal and sham-operated groups,the rate of TUNEL-positive cells and the levels of Bax and cleaved caspase-3 proteins were increased in the spinal cord tissues of rats in the model group(P<0.05),while the expression of Bcl-2 protein was reduced(P<0.05).Compared with the model group,the above indexes were significantly improved in the low-,medium-and high-dose Pueraria lobata decoction groups.(4)Compared with the normal and sham-operated groups,the model group exhibited increased levels of reactive oxygen species,along with elevated expression of NOX4 and p-Drp1 proteins(P<0.05)and reduced expression of Mfn2 protein(P<0.05)in the rat spinal crod tissue.Compared with the model group,the low-,medium-,and high-dose Pueraria lobata decoction groups exhibited reduced levels of reactive oxygen species,as well as decreased expression of NOX4 and p-Drp1 proteins(P<0.05)and increased expression of Mfn2 protein(P<0.05)in the rat spinal cord tissue.To conclude,Pueraria lobata decoction inhibits neuroinflammatory responses and neuronal apoptosis in the rat model of cervical spondylotic myelopathy,and the mechanism of action may be related to the regulation of the NOX4/reactive oxygen species/DRP1 signaling pathway.
10.Regulation of type Ⅰ interferon secretion via the RIG-Ⅰ signaling pathway after PRV infection of mouse trigeminal ganglion cells
Zhengbo LIAO ; Deyuan TANG ; Zhiyong ZENG ; Bin WANG ; Tao HUANG ; Xu CHEN ; Shen-glin YUAN ; Song HE ; Piao ZHOU ; Yinming MAO
Chinese Journal of Veterinary Science 2025;45(2):255-265
This study investigates the effects of pseudorabies virus(PRV)infection on the antiviral immune signaling pathways and type Ⅰ interferon factors in mouse trigeminal ganglion(TG)cells.In this experiment,primary TG cells were infected with PRV at a multiplicity of infection(MOI)of 1,while mice were infected via a drop-nose method using 106,29 TCID50 of PRV.Real-time fluorescence quantitative PCR(qPCR),Western blot and ELISA were used to assess gene tran-scription,protein expression,and the secretion of IFN-α and IFN-β.The results indicated that PRV infection of mouse TG primary cells led to alterations in the gene and protein expression of RIG-Ⅰ,MAVS,and IRF3,as well as the phosphorylation of IRF3 and IKBα both in vivo and ex vivo.ELISA results showed that PRV infection could regulate the secretion of IFN-α and IFN-β in mouse primary TG cells and mouse TGs.The results of RIG-Ⅰ signaling pathway-related proteins and the secretion of IFN-a and IFN-β were analyzed using Western blot after using siRNA to interfere with RIG-Ⅰ expression in TG cells.The results showed that siRIG-Ⅰ successfully inter-fered with RIG-Ⅰ protein expression in TG cells and caused changes in the expression of down-stream proteins such as MAVS and IRF3,and also regulated the secretion of IFN-α and IFN-β in TG cells.Furthermore,the results indicated that PRV infection induced the expression of RIG-Ⅰ in mouse TG progenitor cells,regulating the antiviral immune response of type Ⅰ interferon factors in TG cells through the RIG-Ⅰ-MAVS-IRF3 signaling axis.Notably,PRV inhibited the expression of IRF3 in TG cells while significantly upregulating the expression of IFN-β during the later stages of infection,which may be an important factor in the important reason for the rapid mortality ob-served in mice during the late stages of PRV infection.This experiment elucidates part of the anti-viral immune mechanism mediated by the RIG-Ⅰ-MAVS-IRF3 signaling pathway in regulating type Ⅰ interferon factor after PRV infection of mouse TG cells,as well as the discovery of differ-ent trends of IRF3 protein changes in vivo and ex vivo,laying the groundwork for future in-depth studies.

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