1.Value of three-dimensional inversion-recovery with real reconstruction sequence using an ultralong repetition time for endolymphatic hydrops
Menglong ZHAO ; Huaili JIANG ; Shujie ZHANG ; Zhuang LIU ; Kai LIU ; Di WU ; Xinsheng HUANG ; Mengsu ZENG
Chinese Journal of Clinical Medicine 2025;32(2):200-206
Objective To evaluate the value of an optimized three-dimensional inversion-recovery with real reconstruction (3D-real IR) sequence with a longer repetition time (TR, 16 000 ms) based on modulated flip angle technique in refocused imaging with extended echo train (MATRIX) in the endolymphatic hydrops (EH) imaging after intratympanic gadolinium (Gd) administration, and to compare it with a conventional 3D-real IR based on the turbo spin echo (TSE) sequence. Methods From July 2021 to November 2022, twenty-seven patients received both the conventional and optimized 3D-real IR sequences after bilateral intratympanic Gd administration. Images of the two sequences were qualitativly evaluated and compared. Contrast-to-noise ratio (CNR), signal-to-noise ratio (SNR), and area ratio of endolymph against the total lymphatic space from the two sequences were measured and compared. Results 14(25.9%) ears with insufficient contrast for the EH diagnosis on the conventional sequence were clearly displayed on the optimized sequence. Image score, CNR and SNR of the optimized sequence were significantly higher than those of the conventional sequence (P < 0.001). The scanning time of two sequences was similar. The area ratio of endolymph against the total lymphatic space in the cochlear was significantly higher on the conventional 3D-real IR than that on the optimized 3D-real IR (P < 0.001); there was no statistical difference in the vestibule between the two sequences. Conclusions Compared with conventional sequence, optimized 3D-real IR sequence with a longer TR may be better for evaluation of EH after intratympanic Gd administration.
2.Application of three-dimensional fluid-attenuated inversion recovery sequence using artificial intelligence-assisted compressed sensing technique in intravenous gadolinium contrast-enhanced magnetic resonance imaging of inner ear
Kai LIU ; Jian WANG ; Huaili JIANG ; Shujie ZHANG ; Di WU ; Xinsheng HUANG ; Mengsu ZENG ; Menglong ZHAO
Chinese Journal of Clinical Medicine 2025;32(2):212-217
Objective To investigate the value of artificial intelligence-assisted compressed sensing (ACS) technology for intravenous gadolinium contrast-enhanced magnetic resonance imaging of the inner ear using three-dimensional fluid-attenuated inversion recovery (3D-FLAIR) sequence. Methods The patients received gadolinium contrast-enhanced magnetic resonance imaging using ACS and united compressed sensing (uCS) 3D-FLAIR at Zhongshan Hospital, Fudan University from January to November 2024 were prospectively enrolled. The repetition time was 16 000 ms, and acquisition time was 6 min 40 s and 10 min 24 s in ACS 3D-FLAIR and uCS 3D-FLAIR, respectively. The images on the two sequences were evaluated independently by two radiologists. The image quality of the two sequences was subjectively evaluated and compared. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were compared between the two sequences. The grading consistencies using two sequences and between the two doctors were analyzed. Results There was no statistically difference in subjective score of image quality between the two sequences. SNR and CNR of the ACS 3D-FLAIR sequence were significantly higher than those of the uCS 3D-FLAIR sequence (P<0.001). The kappa values of grades of cochlear and vestibular endolymphatic hydrops were 0.942 and 0.888 using two sequences (P<0.001). The kappa values of grades of cochlear and vestibular endolymphatic hydrops using the ACS 3D-FLAIR sequence between the two doctors were 0.784 and 0.831, respectively (P<0.001); the kappa values of grades of cochlear and vestibular endolymphatic hydrops using uCS 3D-FLAIR sequence between the two doctors were 0.725 and 0.756, respectively (P<0.001). Conclusions ACS 3D-FLAIR could provide higher SNR and CNR than uCS 3D-FLAIR, and is more suitable for intravenous gadolinium contrast-enhanced magnetic resonance imaging of the inner ear; the endolymphatic hydrops grades using ACS 3D-FLAIR is similar to use uCS 3D-FLAIR.
3.2,3,5,4′-tetrahydroxyldiphenylethylene-2-O-glucoside Attenuates Cerebral Ischemia-reperfusion Injury via PINK1/LETM1 Signaling Pathway
Hongyu ZENG ; Kaimei TAN ; Feng QIU ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Chang LEI ; Hongqing ZHAO ; Yuhong WANG ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):145-154
ObjectiveTo investigate the mechanism by which 2,3,5,4'-tetrahydroxyldiphenylethylene-2-O-glucoside (THSG) mitigates cerebral ischemia/reperfusion (CI/R) injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham, model, SAS (40 mg·kg-1), and low-, medium- and high-dose (10, 20, 40 mg·kg-1, respectively) THSG groups, with 10 rats in each group. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring, and cerebral infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 (CCK-8) assay, and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 (PINK1) and leucine zipper/EF-hand-containing transmembrane protein 1 (LETM1) in neurons. Transmission electron microscopy (TEM) was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 (TOMM20), autophagy-associated protein p62, microtubule-associated protein light chain 3 (LC3), cysteinyl aspartate-specific proteinase-9 (Caspase-9), B-cell lymphoma 2-associated protein X (Bax), and cytochrome C (Cyt C). ResultsCompared with the sham group, the model group exhibited increased infarct volume (P<0.01) and neurological deficit scores (P<0.01), neuronal structure was disrupted with reduced Nissl bodies. (P<0.01), mitochondrial swelling/fragmentation, decreased PINK1/LETM1 co-localization (P<0.01), upregulated protein levels of LC3Ⅱ/LC3Ⅰ, TOMM20, Caspase-9, Bax, and Cyt C (P<0.01), downregulated protein level of p62 (P<0.05), weakened PC12 viability (P<0.01), and elevated mitochondrial calcium level (P<0.01). Compared with the model group, THSG and SAS groups showed reduced infarct volumes (P<0.05,P<0.01) and neurological deficit scores (P<0.05,P<0.01), mitigated mitochondrial damage, and increased PINK1/LETM1 co-localization (P<0.01). Medium/high-dose THSG and SAS alleviated the neurological damage, increased Nissl bodies (P<0.05,P<0.01), downregulated the protein levels of p62, TOMM20, Caspase-9, Bax, and Cyt C (P<0.05,P<0.01), and elevated the LC3Ⅱ/LC3Ⅰ level (P<0.05,P<0.01). High-dose THSG enhanced PC12 cell viability (P<0.01), increased PINK1/LETM1 co-localization (P<0.01), and reduced mitochondrial calcium (P<0.01). ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway, reducing the mitochondrial calcium overload, and promoting mitophagy.
4.2,3,5,4′-tetrahydroxyldiphenylethylene-2-O-glucoside Attenuates Cerebral Ischemia-reperfusion Injury via PINK1/LETM1 Signaling Pathway
Hongyu ZENG ; Kaimei TAN ; Feng QIU ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Chang LEI ; Hongqing ZHAO ; Yuhong WANG ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):145-154
ObjectiveTo investigate the mechanism by which 2,3,5,4'-tetrahydroxyldiphenylethylene-2-O-glucoside (THSG) mitigates cerebral ischemia/reperfusion (CI/R) injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham, model, SAS (40 mg·kg-1), and low-, medium- and high-dose (10, 20, 40 mg·kg-1, respectively) THSG groups, with 10 rats in each group. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring, and cerebral infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 (CCK-8) assay, and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 (PINK1) and leucine zipper/EF-hand-containing transmembrane protein 1 (LETM1) in neurons. Transmission electron microscopy (TEM) was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 (TOMM20), autophagy-associated protein p62, microtubule-associated protein light chain 3 (LC3), cysteinyl aspartate-specific proteinase-9 (Caspase-9), B-cell lymphoma 2-associated protein X (Bax), and cytochrome C (Cyt C). ResultsCompared with the sham group, the model group exhibited increased infarct volume (P<0.01) and neurological deficit scores (P<0.01), neuronal structure was disrupted with reduced Nissl bodies. (P<0.01), mitochondrial swelling/fragmentation, decreased PINK1/LETM1 co-localization (P<0.01), upregulated protein levels of LC3Ⅱ/LC3Ⅰ, TOMM20, Caspase-9, Bax, and Cyt C (P<0.01), downregulated protein level of p62 (P<0.05), weakened PC12 viability (P<0.01), and elevated mitochondrial calcium level (P<0.01). Compared with the model group, THSG and SAS groups showed reduced infarct volumes (P<0.05,P<0.01) and neurological deficit scores (P<0.05,P<0.01), mitigated mitochondrial damage, and increased PINK1/LETM1 co-localization (P<0.01). Medium/high-dose THSG and SAS alleviated the neurological damage, increased Nissl bodies (P<0.05,P<0.01), downregulated the protein levels of p62, TOMM20, Caspase-9, Bax, and Cyt C (P<0.05,P<0.01), and elevated the LC3Ⅱ/LC3Ⅰ level (P<0.05,P<0.01). High-dose THSG enhanced PC12 cell viability (P<0.01), increased PINK1/LETM1 co-localization (P<0.01), and reduced mitochondrial calcium (P<0.01). ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway, reducing the mitochondrial calcium overload, and promoting mitophagy.
5.Introduction and enlightenment of the Recommendations and Expert Consensus for Plasm a and Platelet Transfusion Practice in Critically ill Children: from the Transfusion and Anemia Expertise Initiative-Control/Avoidance of Bleeding (TAXI-CAB)
Lu LU ; Jiaohui ZENG ; Hao TANG ; Lan GU ; Junhua ZHANG ; Zhi LIN ; Dan WANG ; Mingyi ZHAO ; Minghua YANG ; Rong HUANG ; Rong GUI
Chinese Journal of Blood Transfusion 2025;38(4):585-594
To guide transfusion practice in critically ill children who often need plasma and platelet transfusions, the Transfusion and Anemia Expertise Initiative-Control/Avoidance of Bleeding (TAXI-CAB) developed Recommendations and Expert Consensus for Plasma and Platelet Transfusion Practice in Critically Ill Children. This guideline addresses 53 recommendations related to plasma and platelet transfusion in critically ill children with 8 kinds of diseases, laboratory testing, selection/treatment of plasma and platelet components, and research priorities. This paper introduces the specific methods and results of the recommendation formation of the guideline.
6.Panax notoginseng saponins regulate differential miRNA expression in osteoclast exosomes and inhibit ferroptosis in osteoblasts
Hongcheng TAO ; Ping ZENG ; Jinfu LIU ; Zhao TIAN ; Qiang DING ; Chaohui LI ; Jianjie WEI ; Hao LI
Chinese Journal of Tissue Engineering Research 2025;29(19):4011-4021
BACKGROUND:Steroid-induced femoral head necrosis is mostly caused by long-term and extensive use of hormones,but its specific pathogenesis is not yet clear and needs further study. OBJECTIVE:To screen out the differential miRNAs in osteoclast exosomes after the intervention of Panax notoginseng saponins,and on this basis,to further construct an osteogenic-related ferroptosis regulatory network to explore the potential mechanism and research direction of steroid-induced osteonecrosis of the femoral head. METHODS:MTT assay was used to detect the toxic effects of different concentrations of dexamethasone and different mass concentrations of Panax notoginseng saponins on Raw264.7 cell line.Tartrate resistant acid phosphatase staining and TUNEL assay were used to detect the effects of Panax notoginseng saponins on osteoclast inhibition and apoptosis.Exosomes were extracted from cultured osteoclasts with Panax notoginseng saponins intervention.Exosomes from different groups were sequenced to identify differentially expressed miRNAs.CytoScape 3.9.1 was used to construct and visualize the regulatory network between differentially expressed miRNAs and mRNAs.Candidate mRNAs were screened by GO analysis and KEGG analysis.Finally,the differential genes related to ferroptosis were screened out,and the regulatory network of ferroptosis-related genes was constructed. RESULTS AND CONCLUSION:(1)The concentration of dexamethasone(0.1 μmol/L)and Panax notoginseng saponins(1 736.85 μg/mL)suitable for intervention of Raw264.7 cells was determined by MTT assay.(2)Panax notoginseng saponins had an inhibitory effect on osteoclasts and could promote their apoptosis.(3)Totally 20 differentially expressed miRNAs were identified from osteoclast-derived exosome samples,and 11 differentially expressed miRNAs related to osteogenesis were predicted by target mRNAs.The regulatory networks of 4 up-regulated differentially expressed miRNAs corresponding to 155 down-regulated candidate mRNAs and 7 down-regulated differentially expressed miRNAs corresponding to 238 up-regulated candidate mRNAs were constructed.(4)Twenty-four genes related to ferroptosis were screened out from the differential genes.Finally,12 networks were constructed(miR-98-5p/PTGS2,miR-23b-3p/PTGS2,miR-425-5p/TFRC,miR-133a-3p/TFRC,miR-185-5p/TFRC,miR-23b-3p/NFE2L2,miR-23b-3p/LAMP2,miR-98-5p/LAMP2,miR-182-5p/LAMP2,miR-182-5p/TLR4,miR-23b-3p/ZFP36,and miR-182-5p/ZFP36).These results indicate that Panax notoginseng saponins may regulate osteoblast ferroptosis by regulating the expression of miRNAs derived from osteoclast exosomes,thus providing a new idea for the study of the mechanism of steroid-induced femoral head necrosis.
7.Unveiling and Validating a Modified Method to Assess Cervical Sagittal Alignment as an Effective Substitute for Conventional C2-7 Cobb Angle
Wenpeng LI ; Qiwei WANG ; Qiancheng ZHAO ; Ziliang ZENG ; Xumin HU ; Xin LV ; Liangbin GAO
Clinics in Orthopedic Surgery 2025;17(1):130-137
Background:
Developing and validating a modified parameter, the SYS-G angle (the angle between the lower endplate of the C2 and the upper endplate of C7 vertebrae), as a feasible substitute for the C2–C7 Cobb method in assessing cervical sagittal alignment and exploring its reference range through a large-scale retrospective study.
Methods:
The visibility of the C6, C7 upper, and C7 lower endplates was graded and compared. Baseline data such as height, weight, body mass index (BMI), age, and sex were analyzed for their impact on the visibility of the C7 lower endplate. Values of C2-6 Cobb angle, SYS-G angle, and C2-7 Cobb angle were measured. The intra- and interobserver reliability, differences, and efficacy of evaluation on cervical lordosis of the parameters were compared, and the correlations among the parameters were analyzed. Furthermore, reference ranges for the SYS-G angle were established based on lateral cervical spine x-rays of 825 asymptomatic Chinese adults across different age groups and sexes.
Results:
The visibility of the C7 lower endplates was significantly reduced compared to the C6 lower and C7 upper endplates.Age, weight, BMI, and male sex were identified as factors negatively influencing the visibility of the C7 lower endplate. Both intraobserver and interobserver reliability demonstrated excellence for all tested parameters. The linear regression model unveiled a stronger association of the SYS-G angle with the C2-7 Cobb angle compared to the C2-6 Cobb angle. Furthermore, the SYS-G angle exhibited excellent efficacy in evaluating cervical lordosis. Age displayed a positive correlation with the SYS-G angle, and across every age bracket from 20 to 69 years, men exhibited a higher mean SYS-G angle compared to women.
Conclusions
The visibility of the C7 lower endplate diminishes with increasing age, weight, BMI, and male sex. In cases where the C7 lower endplate is unclear, the SYS-G angle emerges as a reliable method for estimating cervical sagittal morphology. Reference ranges for the SYS-G angle were established across various age groups and sexes among asymptomatic Chinese adults, offering a valuable resource to guide therapeutic interventions for cervical spine disorders and deformities.
8.Unveiling and Validating a Modified Method to Assess Cervical Sagittal Alignment as an Effective Substitute for Conventional C2-7 Cobb Angle
Wenpeng LI ; Qiwei WANG ; Qiancheng ZHAO ; Ziliang ZENG ; Xumin HU ; Xin LV ; Liangbin GAO
Clinics in Orthopedic Surgery 2025;17(1):130-137
Background:
Developing and validating a modified parameter, the SYS-G angle (the angle between the lower endplate of the C2 and the upper endplate of C7 vertebrae), as a feasible substitute for the C2–C7 Cobb method in assessing cervical sagittal alignment and exploring its reference range through a large-scale retrospective study.
Methods:
The visibility of the C6, C7 upper, and C7 lower endplates was graded and compared. Baseline data such as height, weight, body mass index (BMI), age, and sex were analyzed for their impact on the visibility of the C7 lower endplate. Values of C2-6 Cobb angle, SYS-G angle, and C2-7 Cobb angle were measured. The intra- and interobserver reliability, differences, and efficacy of evaluation on cervical lordosis of the parameters were compared, and the correlations among the parameters were analyzed. Furthermore, reference ranges for the SYS-G angle were established based on lateral cervical spine x-rays of 825 asymptomatic Chinese adults across different age groups and sexes.
Results:
The visibility of the C7 lower endplates was significantly reduced compared to the C6 lower and C7 upper endplates.Age, weight, BMI, and male sex were identified as factors negatively influencing the visibility of the C7 lower endplate. Both intraobserver and interobserver reliability demonstrated excellence for all tested parameters. The linear regression model unveiled a stronger association of the SYS-G angle with the C2-7 Cobb angle compared to the C2-6 Cobb angle. Furthermore, the SYS-G angle exhibited excellent efficacy in evaluating cervical lordosis. Age displayed a positive correlation with the SYS-G angle, and across every age bracket from 20 to 69 years, men exhibited a higher mean SYS-G angle compared to women.
Conclusions
The visibility of the C7 lower endplate diminishes with increasing age, weight, BMI, and male sex. In cases where the C7 lower endplate is unclear, the SYS-G angle emerges as a reliable method for estimating cervical sagittal morphology. Reference ranges for the SYS-G angle were established across various age groups and sexes among asymptomatic Chinese adults, offering a valuable resource to guide therapeutic interventions for cervical spine disorders and deformities.
9.Value of different noninvasive diagnostic models in the diagnosis of esophageal and gastric varices with significant portal hypertension in compensated hepatitis B cirrhosis
Cheng LIU ; Jiayi ZENG ; Mengbing FANG ; Zhiheng CHEN ; Bei GUI ; Fengming ZHAO ; Jingkai YUAN ; Chaozhen ZHANG ; Meijie SHI ; Yubao XIE ; Xiaoling CHI ; Huanming XIAO
Journal of Clinical Hepatology 2025;41(2):263-268
ObjectiveTo investigate the value of different noninvasive diagnostic models in the diagnosis of esophageal and gastric varices since there is a high risk of esophageal and gastric varices in patients with compensated hepatitis B cirrhosis and significant portal hypertension, and to provide a basis for the early diagnosis of esophageal and gastric varices. MethodsA total of 108 patients with significant portal hypertension due to compensated hepatitis B cirrhosis who attended Guangdong Provincial Hospital of Traditional Chinese Medicine from November 2017 to November 2023 were enrolled, and according to the presence or absence of esophageal and gastric varices under gastroscopy, they were divided into esophageal and gastric varices group (GOV group) and non-esophageal and gastric varices group (NGOV group). Related data were collected, including age, sex, imaging findings, and laboratory markers. The chi-square test was used for comparison of categorical data between groups; the least significant difference t-test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups. The receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic value of five scoring models, i.e., fibrosis-4 (FIB-4), LOK index, LPRI, aspartate aminotransferase-to-platelet ratio index (APRI), and aspartate aminotransferase/alanine aminotransferase ratio (AAR). The binary logistic regression method was used to establish a combined model, and the area under the ROC curve (AUC) was compared between the combined model and each scoring model used alone. The Delong test was used to compare the AUC value between any two noninvasive diagnostic models. ResultsThere were 55 patients in the GOV group and 53 patients in the NGOV group. Compared with the NGOV group, the GOV group had a significantly higher age (52.64±1.44 years vs 47.96±1.68 years, t=0.453, P<0.05) and significantly lower levels of alanine aminotransferase [42.00 (24.00 — 17.00) U/L vs 82.00 (46.00 — 271.00) U/L, Z=-3.065, P<0.05], aspartate aminotransferase [44.00 (32.00 — 96.00) U/L vs 62.00 (42.50 — 154.50) U/L,Z=-2.351, P<0.05], and platelet count [100.00 (69.00 — 120.00)×109/L vs 119.00 (108.50 — 140.50)×109/L, Z=-3.667, P<0.05]. The ROC curve analysis showed that FIB-4, LOK index, LPRI, and AAR used alone had an accuracy of 0.667, 0.681, 0.730, and 0.639, respectively, in the diagnosis of esophageal and gastric varices (all P<0.05), and the positive diagnostic rates of GOV were 69.97%, 65.28%, 67.33%, and 58.86%, respectively, with no significant differences in AUC values (all P>0.05), while APRI used alone had no diagnostic value (P>0.05). A combined model (LAF) was established based on the binary logistic regression analysis and had an AUC of 0.805 and a positive diagnostic rate of GOV of 75.80%, with a significantly higher AUC than FIB-4, LOK index, LPRI, and AAR used alone (Z=-2.773,-2.479,-2.206, and-2.672, all P<0.05). ConclusionFIB-4, LOK index, LPRI, and AAR have a similar diagnostic value for esophageal and gastric varices in patients with compensated hepatitis B cirrhosis and significant portal hypertension, and APRI alone has no diagnostic value. The combined model LAF had the best diagnostic efficacy, which provides a certain reference for clinical promotion and application.
10.Effects of Electroacupuncture at "Fengchi" (GB 20), "Waiguan" (TE 5), and "Yanglingquan" (GB 34) on Nociceptive Sensitization and PKC/TRPV1 Pathway in the Trigeminal Ganglion of Chronic Migraine Model Rats
Yixiang ZENG ; Runze TU ; Shucong ZHAO ; Yang YANG ; Haojia WEN ; Zhuozhong HE ; Shengli ZHOU ; Lei TAN ; Ke HE ; Lei FU
Journal of Traditional Chinese Medicine 2025;66(3):283-289
ObjectiveTo explore the possible mechanisms of electroacupuncture at Fengchi (GB 20), Waiguan (TE 5), and Yanglingquan (GB 34) in treating chronic migraine from the perspective of nociceptive sensitization. MethodsForty SPF-grade SD rats were randomly divided into blank group, model group, electroacupuncture group, electroacupuncture + agonist group, and inhibitor group, with 8 rats in each group. Except for the blank group, rats were injected intraperitoneally with nitroglycerin to establish a chronic migraine rat model. After successful modeling, the electroacupuncture group received electroacupuncture at bilateral "Fengchi" (GB 20), "Waiguan" (TE 5), and "Yanglingquan" (GB 34) for 30 minutes each session. The electroacupuncture + agonist group received the same electroacupuncture treatment and additional injection of protein kinase C (PKC) agonist Phorbol 12-myristate 13-acetate (1.0 ng/μl, 25 μl) via the infraorbital foramen. The inhibitor group received PKC inhibitor Chelerythrine Chloride (1.0 ng/μl, 10 μl) via the infraorbital foramen. The blank group, model group, and inhibitor group underwent restraint for 30 minutes without other interventions. All groups were continuously intervened for 5 days. After the intervention, the nociceptive thresholds (mechanical and thermal pain) of the periorbital area and hind paw were measured. The expression levels of transient receptor potential vanillic acid subtype 1 (TRPV1), phosphorylated TRPV1 (p-TRPV1), PKC proteins, Trpv1, Pkc mRNA, and the average fluorescence intensity of transient receptor potential vanillic acid subtype 1 (TRPV1) and PKC in the trigeminal ganglion were detected using Western Blot, real-time fluorescence quantitative PCR, and immunofluorescence methods. ResultsCompared with the blank group, the mechanical and thermal pain thresholds of the periorbital area and hind paw were reduced in the model group, and the protein levels of TRPV1, PKC, p-TRPV1, as well as the mRNA expression of Trpv1 and Pkc, and the average fluorescence intensity of TRPV1 and PKC in the trigeminal ganglion significantly increased (P<0.05 or P<0.01). Compared with the model group, the electroacupuncture group exhibited increased mechanical and thermal pain thresholds in the periorbital and hind paw areas, and decreased protein levels of TRPV1, PKC, p-TRPV1, mRNA expression of Trpv1 and Pkc, and average fluorescence intensity of TRPV1. In the electroacupuncture + agonist group, the average fluorescence intensity of TRPV1 in the trigeminal ganglion decreased. The inhibitor group exhibited increased mechanical pain thresholds in the periorbital area and thermal pain thresholds in the hind paw, along with decreased protein levels of TRPV1, PKC, p-TRPV1, and the average fluorescence intensity of TRPV1 and PKC (P<0.05 or P<0.01). Compared with the electroacupuncture group, the electroacupuncture + agonist group showed an increase in the protein levels of TRPV1, PKC, p-TRPV1, and the mRNA expression of Trpv1 (P<0.05 or P<0.01). ConclusionElectroacupuncture at the "Fengchi" (GB 20), "Waiguan" (TE 5), and "Yanglingquan" (GB 34) acupoints can increase the mechanical and thermal pain thresholds in chronic migraine rats and alleviate nociceptive sensitization. The mechanism may be related to the inhibition of PKC/TRPV1 pathway.

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