1.Differences in deltamethrin resistance and kdr gene mutation in Culex tritaeniorhynchus population in and outside the Yellow Sea wetland
Xiao-er ZHANG ; Zhi-ming WU ; Ye TIAN ; Qian CUI ; Yu-qian JI ; Huan WANG ; Shu-juan YANG ; Yi-chao ZHAO ; Yu WANG ; Hua-yu YIN ; Yu DING ; Guo-jin YAN ; Min-sen ZHAO ; Shou-gang ZHANG ; Bing-dong SONG ; Hong-na CHEN ; Jian GAO ; Wei-fang YANG ; Yu-fu ZHANG ; Hui LIU ; Hong-liang CHU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):101-107
Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.
2.Transcranial Focused Ultrasound Stimulation of the Primary Somatosensory Cortex Evokes Hand Tactile Perception: The Effect of Sonication Duration
Xu-Xu CHEN ; Huan-Jun XI ; Zhen LIANG ; Bo WANG ; Xu-Dong ZHAO
Progress in Biochemistry and Biophysics 2026;53(7):1942-1952
ObjectiveTranscranial focused ultrasound (tFUS) has emerged as a promising noninvasive neuromodulation technique capable of modulating neural activity with high spatial specificity. However, whether tFUS stimulation of the primary somatosensory cortex (S1) can reliably evoke conscious tactile perception without external peripheral sensory input remains unclear. Therefore, this study examined the feasibility of inducing hand-related tactile perception through targeted S1 stimulation and further investigated how sonication duration (SD), a key temporal parameter, influences the consistency of this perceptual response. The findings may provide empirical evidence for optimizing tFUS parameters in human somatosensory modulation. MethodsForty-eight healthy adults participated in the study and were assigned to two experiments. Experiment 1 included 24 participants and was designed to preliminarily examine whether S1-targeted tFUS could evoke hand-related tactile sensations. This experiment comprised two sub-experiments, Experiment 1a and Experiment 1b, with 12 participants in each. Specifically, Experiment 1a used a randomized active stimulation protocol targeting the bilateral S1 to assess the initial feasibility of inducing contralateral tactile perception, whereas Experiment 1b employed an alternating hemispheric design incorporating both active and sham stimulation to verify the specificity of the tFUS-induced sensations. Experiment 2 enrolled another 24 participants and used a real-sham dual-probe design to control for spatial tactile cues that might arise from probe-scalp contact. Five SD levels, including 200, 400, 600, 800, and 1 000 ms, were systematically tested to evaluate how stimulation duration influenced the behavioral response. The primary behavioral outcome was the perceptual consistency rate, quantified according to the accuracy of contralateral tactile localization. ResultsIn Experiment 1, 91.7% of participants reported reliable hand-related tactile sensations during active tFUS stimulation, including numbness, tingling, or similar sensory experiences. Contralateral localization accuracy under active stimulation was significantly higher than the0.50 chance level (P<0.05). In contrast, localization performance under sham stimulation did not differ from chance, suggesting that the observed tactile responses were mainly induced by active S1-targeted tFUS stimulation rather than nonspecific stimulation cues. In Experiment 2, perceptual consistency rates significantly exceeded the chance level under the 400 and 1 000 ms SD conditions(P<0.05), whereas no significant effect was observed at 200, 600, or 800 ms. Pairwise comparisons further showed that the 400 and 1 000 ms conditions produced significantly higher perceptual consistency rates than the 200 ms condition (P<0.05). ConclusiontFUS stimulation of S1 can elicit distinct hand-related tactile sensations without peripheral sensory input, demonstrating the feasibility of using targeted cortical ultrasound stimulation to induce conscious somatosensory perception. The reliability of this perceptual effect was modulated by sonication duration, indicating that temporal stimulation parameters play an important role in shaping both the occurrence and consistency of behavioral responses. Among the tested conditions, 400 ms appeared to provide a favorable temporal window for generating stable tactile perception, whereas excessively short stimulation may be insufficient to produce consistent responses. These findings highlight the sensitivity of somatosensory networks to ultrasonic modulation and provide useful parameter guidance for the application of tFUS in noninvasive sensory enhancement and cortical functional mapping.
3.Progress on M cells-targeted oral vaccine delivery systems
Anze LIU ; Yingxin DONG ; Huan WANG ; Ying LU
Journal of Pharmaceutical Practice and Service 2026;44(8):389-394
Oral vaccines, owing to their non-invasive nature, ease of administration, and ability to elicit both mucosal and systemic immune responses, represent a pivotal direction in vaccine development. Microfold cells (M cells), a specialized subset of immune-surveillance epithelial cells, though sparsely distributed, possess exceptional capacity for antigen uptake and transcytosis, rendering them a critical target for overcoming the bottlenecks of oral vaccine delivery. The morphological structure, antigen uptake mechanisms, and surface receptor characteristics of intestinal M cells were summarized in this paper. The multifaceted challenges currently impeding M cell-targeted oral delivery and recent advances in targeting strategies based on receptor–ligand specific recognition were analyzed. The design and application of delivery carriers tailored for M cell targeting were reviewed, which aimed to provide insights and guidance for future research and development of M cell-targeted mucosal delivery systems in the intestine.
4.Mechanism of Shengxian Yixin Granule (升陷益心颗粒) in the Treatment of Heart Failure with Preserved Ejection Fraction Based on Macrophage-Mediated Inflammatory Response
Zhengwei DONG ; Rui YU ; Qiaozhi LI ; Huan ZHAO ; Jingjing WEI ; Genhao FAN ; Xinlu WANG ; He WANG ; Yongxia WANG ; Mingjun ZHU
Journal of Traditional Chinese Medicine 2026;67(19):2086-2094
ObjectiveTo investigate the effect and mechanism of Shengxian Yixin Granule (升陷益心颗粒, SYG) in treating heart failure with preserved ejection fraction (HFpEF). MethodsExperiment 1: Forty male C57BL/6 mice were randomly divided into control group, model group, low-dose [25 mg/(kg·d)], medium-dose [50 mg/(kg·d)], and high-dose [(75 mg/(kg·d)] SYG groups, with 8 mice per group. Except for the control group, HFpEF was induced by a high-fat diet combined with Nω-nitro-L-arginine methyl ester in drinking water for 16 weeks. After 10 weeks of modeling, treatment groups received corresponding doses of SYG by gavage, while the control group and model group received equal volume of normal saline, for 6 weeks. After 6 weeks of administration, the body weight, blood pressure including systolic blood pressure (SBP) and diastolic blood pressure (DBP), and heart weight-to-tibia length ratio (HW/TL) were recorded. Echocardiography was used to evaluate left ventricular ejection fraction (LVEF) and the ratio of early to late diastolic transmitral flow velocity (E/A). Plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) and C-reactive protein (CRP) levels were measured by ELISA. The expression level of brain natriuretic peptide (BNP) in heart tissue was detected by qRT-PCR. Blood lipid levels, including low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), and total cholesterol (TC), were determined by a biochemical analyzer. Splenic proportions of M1 macrophages, M2 macrophages, T helper 17 (Th17) cells, and regulatory T (Treg) cells were analyzed by flow cytometry. Experiment 2: RAW 264.7 cells were stimulated with lipopolysaccharide (LPS) to establish an M1 macrophage polarization model, and then treated with SYG-containing serum for 24 h. The cells were divided into four groups, including blank serum group, high-dose SYG-containing serum group, LPS group, and LPS plus high-dose SYG-containing serum group. Combined with asignal transducer and activator of transcription 3 (Stat3) inhibitor for 24 h, the cells were divided into a lipopolysaccharide group, a high-dose SYG-containing serum group, an inhibitor group, and high-dose SYG-containing serum plus inhibitor group. The expression levels of inducible nitric oxide synthase (iNOS) and interleukin-1β (IL-1β) in each group were detected by qRT-PCR. The protein levels of iNOS, IL-1β, Stat3, and phosphorylated Stat3 (p-Stat3) were detected by Western Blotting. ResultsExperiment 1: Compared to the control group, the model group showed significantly decreased E/A ratio, and increased cardiac BNP expression, plasma NT-proBNP,HW/TL ratio, body weight, TC, LDL-C, SBP, and DBP, as well as shortened exhaustion distance and exhaustion time (P<0.05 or P<0.01). Compared to the model group, the E/A ratio increased in the low-, medium-, and high-dose SYG groups; cardiac BNP expression decreased in the low- and high-dose groups, and plasma NT-proBNP levels decreased in the high-dose group; the HW/TL ratio and body weight decreased, while exhaustion distance and exhaustion time increased in the medium- and high-dose groups; TG levels decreased in the low- and medium-dose groups, and LDL-C levels decreased in the medium- and high-dose groups; SBP and DBP decreased in the low-, medium-, and high-dose groups (P<0.05 or P<0.01). Compared to the low-dose SYG group, body weight decreased, and both exhaustion distance and exhaustion time increased in the medium- and high-dose groups (P<0.05 or P<0.01). Compared to the medium-dose SYG group, the exhaustion time increased in the high-dose SYG group (P<0.01). Experiment 2: In the LPS‑induced M1 macrophage polarization model, compared to the blank serum group, the LPS group showed significantly increased mRNA and protein expression of iNOS and IL‑1β, as well as the p‑Stat3/Stat3 ratio (P<0.01). Compared to the LPS group, the high‑dose SYG-containing serum plus LPS group showed significantly decreased mRNA and protein expression of iNOS and IL‑1β, as well as the p‑Stat3/Stat3 ratio (P<0.05 or P<0.01). Compared to the LPS group, the inhibitor group also showed significantly reduced these parameters (P<0.05 or P<0.01). ConclusionSYG can improve cardiac function, ventricular remodeling, exercise capacity, and lipid metabolism in HFpEF mice, while inhibiting inflammatory responses. Its mechanism may be related to the regulation of the Stat3 signaling pathway and inhibition of M1 macrophage polarization.
5.Mechanism of Shengxian Yixin Granule (升陷益心颗粒) in the Treatment of Heart Failure with Preserved Ejection Fraction Based on Macrophage-Mediated Inflammatory Response
Zhengwei DONG ; Rui YU ; Qiaozhi LI ; Huan ZHAO ; Jingjing WEI ; Genhao FAN ; Xinlu WANG ; He WANG ; Yongxia WANG ; Mingjun ZHU
Journal of Traditional Chinese Medicine 2026;67(19):2086-2094
ObjectiveTo investigate the effect and mechanism of Shengxian Yixin Granule (升陷益心颗粒, SYG) in treating heart failure with preserved ejection fraction (HFpEF). MethodsExperiment 1: Forty male C57BL/6 mice were randomly divided into control group, model group, low-dose [25 mg/(kg·d)], medium-dose [50 mg/(kg·d)], and high-dose [(75 mg/(kg·d)] SYG groups, with 8 mice per group. Except for the control group, HFpEF was induced by a high-fat diet combined with Nω-nitro-L-arginine methyl ester in drinking water for 16 weeks. After 10 weeks of modeling, treatment groups received corresponding doses of SYG by gavage, while the control group and model group received equal volume of normal saline, for 6 weeks. After 6 weeks of administration, the body weight, blood pressure including systolic blood pressure (SBP) and diastolic blood pressure (DBP), and heart weight-to-tibia length ratio (HW/TL) were recorded. Echocardiography was used to evaluate left ventricular ejection fraction (LVEF) and the ratio of early to late diastolic transmitral flow velocity (E/A). Plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) and C-reactive protein (CRP) levels were measured by ELISA. The expression level of brain natriuretic peptide (BNP) in heart tissue was detected by qRT-PCR. Blood lipid levels, including low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), and total cholesterol (TC), were determined by a biochemical analyzer. Splenic proportions of M1 macrophages, M2 macrophages, T helper 17 (Th17) cells, and regulatory T (Treg) cells were analyzed by flow cytometry. Experiment 2: RAW 264.7 cells were stimulated with lipopolysaccharide (LPS) to establish an M1 macrophage polarization model, and then treated with SYG-containing serum for 24 h. The cells were divided into four groups, including blank serum group, high-dose SYG-containing serum group, LPS group, and LPS plus high-dose SYG-containing serum group. Combined with asignal transducer and activator of transcription 3 (Stat3) inhibitor for 24 h, the cells were divided into a lipopolysaccharide group, a high-dose SYG-containing serum group, an inhibitor group, and high-dose SYG-containing serum plus inhibitor group. The expression levels of inducible nitric oxide synthase (iNOS) and interleukin-1β (IL-1β) in each group were detected by qRT-PCR. The protein levels of iNOS, IL-1β, Stat3, and phosphorylated Stat3 (p-Stat3) were detected by Western Blotting. ResultsExperiment 1: Compared to the control group, the model group showed significantly decreased E/A ratio, and increased cardiac BNP expression, plasma NT-proBNP,HW/TL ratio, body weight, TC, LDL-C, SBP, and DBP, as well as shortened exhaustion distance and exhaustion time (P<0.05 or P<0.01). Compared to the model group, the E/A ratio increased in the low-, medium-, and high-dose SYG groups; cardiac BNP expression decreased in the low- and high-dose groups, and plasma NT-proBNP levels decreased in the high-dose group; the HW/TL ratio and body weight decreased, while exhaustion distance and exhaustion time increased in the medium- and high-dose groups; TG levels decreased in the low- and medium-dose groups, and LDL-C levels decreased in the medium- and high-dose groups; SBP and DBP decreased in the low-, medium-, and high-dose groups (P<0.05 or P<0.01). Compared to the low-dose SYG group, body weight decreased, and both exhaustion distance and exhaustion time increased in the medium- and high-dose groups (P<0.05 or P<0.01). Compared to the medium-dose SYG group, the exhaustion time increased in the high-dose SYG group (P<0.01). Experiment 2: In the LPS‑induced M1 macrophage polarization model, compared to the blank serum group, the LPS group showed significantly increased mRNA and protein expression of iNOS and IL‑1β, as well as the p‑Stat3/Stat3 ratio (P<0.01). Compared to the LPS group, the high‑dose SYG-containing serum plus LPS group showed significantly decreased mRNA and protein expression of iNOS and IL‑1β, as well as the p‑Stat3/Stat3 ratio (P<0.05 or P<0.01). Compared to the LPS group, the inhibitor group also showed significantly reduced these parameters (P<0.05 or P<0.01). ConclusionSYG can improve cardiac function, ventricular remodeling, exercise capacity, and lipid metabolism in HFpEF mice, while inhibiting inflammatory responses. Its mechanism may be related to the regulation of the Stat3 signaling pathway and inhibition of M1 macrophage polarization.
6.Influencing factors for recompensation in patients with decompensated hepatitis C cirrhosis
Danqing XU ; Huan MU ; Yingyuan ZHANG ; Lixian CHANG ; Yuanzhen WANG ; Weikun LI ; Zhijian DONG ; Lihua ZHANG ; Yijing CHENG ; Li LIU
Journal of Clinical Hepatology 2025;41(2):269-276
ObjectiveTo investigate the influencing factors for recompensation in patients with decompensated hepatitis C cirrhosis, and to establish a predictive model. MethodsA total of 217 patients who were diagnosed with decompensated hepatitis C cirrhosis and were admitted to The Third People’s Hospital of Kunming l from January, 2019 to December, 2022 were enrolled, among whom 63 patients who were readmitted within at least 1 year and had no portal hypertension-related complications were enrolled as recompensation group, and 154 patients without recompensation were enrolled as control group. Related clinical data were collected, and univariate and multivariate analyses were performed for the factors that may affect the occurrence of recompensation. The independent-samples t test was used for comparison of normally distributed measurement data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed measurement data between two groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups. A binary Logistic regression analysis was used to investigate the influencing factors for recompensation in patients with decompensated hepatitis C cirrhosis, and the receiver operating characteristic (ROC) curve was used to assess the predictive performance of the model. ResultsAmong the 217 patients with decompensated hepatitis C cirrhosis, 63 (29.03%) had recompensation. There were significant differences between the recompensation group and the control group in HIV history (χ2=4.566, P=0.034), history of partial splenic embolism (χ2=6.687, P=0.014), Child-Pugh classification (χ2=11.978, P=0.003), grade of ascites (χ2=14.229, P<0.001), albumin (t=4.063, P<0.001), prealbumin (Z=-3.077, P=0.002), high-density lipoprotein (t=2.854, P=0.011), high-sensitivity C-reactive protein (Z=-2.447, P=0.014), prothrombin time (Z=-2.441, P=0.015), carcinoembryonic antigen (Z=-2.113, P=0.035), alpha-fetoprotein (AFP) (Z=-2.063, P=0.039), CA125 (Z=-2.270, P=0.023), TT3 (Z=-3.304, P<0.001), TT4 (Z=-2.221, P=0.026), CD45+ (Z=-2.278, P=0.023), interleukin-5 (Z=-2.845, P=0.004), tumor necrosis factor-α (Z=-2.176, P=0.030), and portal vein width (Z=-5.283, P=0.005). The multivariate analysis showed that history of partial splenic embolism (odds ratio [OR]=3.064, P=0.049), HIV history (OR=0.195, P=0.027), a small amount of ascites (OR=3.390, P=0.017), AFP (OR=1.003, P=0.004), and portal vein width (OR=0.600, P<0.001) were independent influencing factors for the occurrence of recompensation in patients with decompensated hepatitis C cirrhosis. The ROC curve analysis showed that HIV history, grade of ascites, history of partial splenic embolism, AFP, portal vein width, and the combined predictive model of these indices had an area under the ROC curve of 0.556, 0.641, 0.560, 0.589, 0.745, and 0.817, respectively. ConclusionFor patients with decompensated hepatitis C cirrhosis, those with a history of partial splenic embolism, a small amount of ascites, and an increase in AFP level are more likely to experience recompensation, while those with a history of HIV and an increase in portal vein width are less likely to experience recompensation.
7.Anti-frostbite effect of miglitol on cold-exposed mice through UCP1-mediated thermogenic activation
Xiang LI ; Hongyuan LU ; Mingyu ZHANG ; Huan GAO ; Dong YAO ; Zihua XU
Journal of Pharmaceutical Practice and Service 2025;43(1):1-5
Objective To investigate the effect and mechanism of miglitol on regulating the energy metabolism of brown adipocytes by activating UCP1 and preventing cold injury in mice after cold exposure. Methods Primary brown adipocytes were induced into mature adipocytes, the effect of miglitol on the viability of brown adipocytes was investigated by MTT method, the lipid droplet consumption level of cells after drug administration was investigated by Oil Red O staining technology, and the level of UCP1, a key protein of thermogenesis in brown adipocytes, was detected by Western blotting. The activity of anti-frostbite was investigated in cold exposure at 4 ℃ and −20 ℃. KM mice, which were randomly divided into control group, cold exposure group, miglitol group and all-trans retinoic acid group, and after 7 days of repeated administration, the body surface temperature of mice was detected by infrared thermal imaging system, the anal temperature change was detected by anal thermometer, and the expression levels of UCP1 and PGC1-α in adipose tissue were detected by immunoblotting. Results Compared with the control group, the lipid droplet consumption and UCP1 expression levels in brown adipocytes in the miglitol group were significantly increased. The levels of body surface temperature and rectal temperature increased significantly after cold exposure, and the levels of UCP1 and PGC1α in the brown adipose tissue of mice increased significantly, which indicated that the miglitol could activate the critical proteins UCP1 and PGC1α of the thermogenesis pathway, increase the thermogenesis of mice after cold exposure, and thus improve the effect of cold injury for toe swelling. Conclusion Miglitol could play a role in improving cold injury and body temperature in mice by increasing the level of UCP1 and PGC1α, which are key targets of the thermogenesis pathway to promote the thermogenesis of brown fat.
8.Manufacture and mechanical property on zirconia abutments with a titanium base in dental implant restoration
Huan WANG ; Jing LU ; Ying LI ; Maohua MENG ; Jiayu SHU ; Yuncai LUO ; Wenjie LI ; Qiang DONG
Chinese Journal of Tissue Engineering Research 2025;29(10):2171-2177
BACKGROUND:With the development of computer-aided design and computer-aided manufacturing technology,zirconia abutments with a titanium base are widely used in clinic due to its good application advantages,but there are still some problems and a lack of consensus design standards. OBJECTIVE:To review the fabrication methods of Ti-base zirconia abutment,and the effect of abutment connection,emergence design,abutment angle,and bonding on mechanical properties of Ti-base zirconia abutment. METHODS:Relevant literature published from 2010 to 2023 was searched in CNKI and PubMed databases with the search terms"zirconia abutment,titanium base"in Chinese and English,respectively.The search time limit was extended for some classical literature.The relevant literature was obtained through inclusion and exclusion criteria,and 57 eligible documents were included for review. RESULTS AND CONCLUSION:It is recommended that clinicians try to select antirotational titanium bases or rotational titanium bases with a Morse taper connection.Implants should be placed in the correct axial angulation of not more than 15° or with an inclination to the palatal side when using angled zirconia abutments.When a≥30° labial inclination is followed for implant placement,the bite force must be decreased effectively to reduce the risk of mechanical and biological complications of implants,abutments,and prostheses.Ti-base zirconia abutments with a higher gingival height should be selected,and its restorative angle should not exceed 40°.Multilink Hybrid Abutment could be the first choice for extraoral bonding of zirconia abutment to titanium bases.
9.Efficacy and mechanism of Guizhi Tongluo Tablets in alleviating atherosclerosis by inhibiting CD72hi macrophages.
Xing-Ling HE ; Si-Jing LI ; Zi-Ru LI ; Dong-Hua LIU ; Xiao-Jiao ZHANG ; Huan HE ; Xiao-Ming DONG ; Wen-Jie LONG ; Wei-Wei ZHANG ; Hui-Li LIAO ; Lu LU ; Zhong-Qi YANG ; Shi-Hao NI
China Journal of Chinese Materia Medica 2025;50(5):1298-1309
This study investigates the effect and underlying mechanism of Guizhi Tongluo Tablets(GZTL) in treating atherosclerosis(AS) in a mouse model. Apolipoprotein E-knockout(ApoE~(-/-)) mice were randomly assigned to the following groups: model, high-, medium-, and low-dose GZTL, and atorvastatin(ATV), and age-matched C57BL/6J mice were selected as the control group. ApoE~(-/-) mice in other groups except the control group were fed with a high-fat diet for the modeling of AS and administrated with corresponding drugs via gavage for 8 weeks. General conditions, signs of blood stasis, and body mass of mice were monitored. Aortic plaques and their stability were assessed by hematoxylin-eosin, Masson, and oil red O staining. Serum levels of total cholesterol(TC), triglycerides(TG), and low-density lipoprotein cholesterol(LDL-C) were measured by biochemical assays, and those of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6) were determined via enzyme-linked immunosorbent assay. Apoptosis was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL). Single-cell RNA sequencing(scRNA-seq) was employed to analyze the differential expression of CD72hi macrophages(CD72hi-Mφ) in the aortas of AS patients and mice. The immunofluorescence assay was employed to visualize CD72hi-Mφ expression in mouse aortic plaques, and real-time fluorescence quantitative PCR was utilized to determine the mRNA levels of IL-1β, TNF-α, and IL-6 in the aorta. The results demonstrated that compared with the control group, the model group exhibited significant increases in body mass, aortic plaque area proportion, necrotic core area proportion, and lipid deposition, a notable decrease in collagen fiber content, and an increase in apoptosis. Additionally, the model group showcased elevated serum levels of TC, TG, LDL-C, IL-1β, TNF-α, and IL-6, alongside marked upregulations in the mRNA levels of IL-1β, TNF-α, and IL-6 in the aorta. In comparison with the model group, the GZTL groups and the ATV group showed a reduction in body mass, and the medium-and high-dose GZTL groups and the ATV group demonstrated reductions in aortic plaque area proportion, necrotic core area proportion, and lipid deposition, an increase in collagen fiber content, and a decrease in apoptosis. Furthermore, the treatment goups showcased lowered serum levels of TC, TG, LDL-C, IL-1β, TNF-α, and IL-6. The data of scRNA-seq revealed significantly elevated CD72hi-Mφ signaling in carotid plaques of AS patients compared with that in the normal arterial tissue. Animal experiments confirmed that CD72hi-Mφ expression, along with several pro-inflammatory cytokines, was significantly upregulated in the aortas of AS mice, which were downregulated by GZTL treatment. In conclusion, GZTL may alleviate AS by inhibiting CD72hi-Mφ activity.
Animals
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Drugs, Chinese Herbal/administration & dosage*
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Atherosclerosis/immunology*
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Mice
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Mice, Inbred C57BL
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Macrophages/immunology*
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Male
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Humans
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Apolipoproteins E/genetics*
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Tablets
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Tumor Necrosis Factor-alpha/genetics*
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Apoptosis/drug effects*
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Interleukin-1beta/genetics*
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Interleukin-6/genetics*
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Disease Models, Animal
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Mice, Knockout
10.miR-21-5p Targetes STAT3 Reduce the OGD/R-induced Neuronal Injury
Yanping LI ; Qingyun LI ; Rong HUO ; Xiaolin DONG ; Hongmei LI ; Huan WEI ; Yi ZENG
Journal of Kunming Medical University 2025;46(5):12-20
Objective To investigate the potential mechanism of miR-21-5p in alleviating cerebral ischemia-reperfusion injury by targeting STAT3.Methods The HT22 cells were induced by OGD/R to construct a cell model of cerebral ischemia reperfusion injury.The expression of miR-21-5p was detected by RT-qPCR.The CCK-8 assay,TUNEL staining and flow cytometry were respectively used to detect the cell viability and apoptosis.ELISA assay was used to determine the contents of inflammatory factors IL-6,IL-10 and TNF-α in the cell supernatant.Western blot was used to detect the expression levels of p-STAT3/STAT3,Cleaved-Caspase-3,Bax and Bcl-2 proteins.The TargetScan database was used to predict the binding sites of miR-21-5p and STAT3.The dual-luciferase reporter gene assay was used to verify the targeting relationship between miR-21-5p and STAT3.Results The relative expression level of miR-21-5p was down-regulated in HT22 cells which induced by OGD/R(P<0.001).The cell viability(P<0.0001)was decreased and the apoptosis rate(P<0.001)was increased in OGD/R induced-HT22 cells.The contents of pro-inflammatory factors IL-6(P<0.001)and TNF-α(P<0.001)was increased,while the content of anti-inflammatory factor IL-10(P<0.001)decreased.After transfection with miR-21-5p mimic,cell viability was enhanced,apoptosis rate was reduced and neuroinflammation was inhibited.MiR-21-5p could target and bind to STAT3.After miR-21-5p inhibitor transfection,cell viability decreased,apoptosis was promoted,and neuroinflammation occurred;STAT3 inhibitor Stattic could reverse the effect of miR-21-5p inhibitor.Conclusion MiR-21-5p could specifically bind to STAT3 and reduce the neuroinflammation and apoptosis of OGD/R induced-HT22 cells.


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