1.Ameliorating effect of betaine on oxygen-glucose deprivation injury in rat brain microvascular endothelial cells and its influence in PI3K/AKT pathway
Min CHEN ; Huiyan ZHU ; Jing TAO ; Yipeng XU
Journal of Jilin University(Medicine Edition) 2025;51(1):96-104
Objective:To investigate the effect of betaine in oxygen-glucose deprivation injury of rat brain microvascular endothelial cells(BMECs),and to clarify the regulatory effect of betaine on phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathway.Methods:Five SD rats aged 7 d were selected and the rat BMSEs were obtained.The oxygen-glucose deprivation model of rat BMECs was prepared under hypoxic and hypoglycemic conditions;the experiment was divdided into model group,and low dose,medium dose,and high dose of betaine groups and positive control group,at the same time,blank control group(without modeling)was set up.The BMECs in blank control group and model group were treated with fresh medium,the BMECs in positive control group were given a final concentration of 10 μmnol·L-1 nimodipine,and the BMECs in low,medium and high doses of betaine groups were treated with betaine at final concentration of 0.5,1.0 and 2.0 mmol·L-1,respectively.The survival rates of BMECs in various groups were determined by CCK-8 method at 12,24 and 48 h after culture;the activities of lactate dehydrogenase(LDH)and the levels of adenine ribonucleoside triphosphate(ATP)in the rat BMECs in various groups were determined using kits,and the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1β,and IL-18 in supernatants of the BMECs in various groups were determined by enzyme-linked immunosorbent assay(ELISA);the activities of superoxide dismutase(SOD)and levels of malondialdehyde(MDA)in the BMECs in various groups were determined by kits;the transendothelial resistance(TEER)values of rat BMSCs in various groups were determined by TEER analyzer,and the horseradish peroxidase(HRP)permeabilities of BMECs in various groups were determined by an insertion cell culture apparatus.TUNEL staining was used to determine the apoptotic rates of rat BMECs in vaisous groups,and Western blotting method was used to determine the ratios of phosphory lated PI3K(p-PI3K)/PI3K and phosphorylated AKT(p-AKT)/AKT in the rat BMECs in various groups.Results:Compared with blank control group,the survival rate of BMECs,activity of SOD,and level of ATP,value of TEER,and ratios of p-PI3K/PI3K and p-AKT/AKT of the rat BMECs in model group were significantly decreased(P<0.05),while the activity of LDH,the levels of TNF-α,IL-6,IL-1β,IL-18,and MDA,the apoptotic rate of the BMECs,and HRP permeability were significantly increased(P<0.05).Compared with model group,the survival rates of the BMECs,activities of SOD,and levels of ATP,values of TEER,and ratios of p-PI3K/PI3K and p-AKT/AKT of the BMECs in low,medium,and high doses of betaine groups and positive control group were significantly increased(P<0.05),while the activities of LDH,the levels of TNF-α,IL-6,IL-1β,IL-18,and MDA,the apoptotic rates of BMECs and HRP permeabilities were significantly decreased(P<0.05).Conclusion:Betaine can significantly repair the oxygen-glucose deprivation/reperfusion injury in the rat BMECs,inhibit the oxidative damage and apoptosis of BMECs,and improve the permeability of the cells;its mechanism may be related to the regulation of the PI3K/AKT pathway.
2.α-mangostin inhibits LPS/ATP-induced NLRP3 inflammasome activation in microglia via the NF- κB pathway
Min Chen ; Jing Tao ; Huiyan Zhu
Acta Universitatis Medicinalis Anhui 2024;59(4):575-580
Objective :
To investigate the effects and underlying mechanisms of α-mangostin in a spinal cord inj ury model of microglial cell inflammation .
Methods :
Mouse microglial cell line BV-2 was cultured in vitro , and an in- flammation model was established by co-treatment with lipopolysaccharide and adenosine triphosphate (LPS/ATP) . The CCK-8 assay was used to test the influence of different concentrations (0 , 10 , 20 , 40 , 80 μmol/L) of α-man- gostin on cell proliferation vitality under LPS/ATP stimulation to select an appropriate concentration range of α- mangostin; BV-2 cells were divided into Ctrl group , LPS/ATP group , 40 μmol/L α-mangostin group , and inter- vention groups with different concentrations (10 , 20 , 40 μmol/L) of α-mangostin ( designated as LPS/ATP + 10 μmol/L α-mangostin group , LPS/ATP + 20 μmol/L α-mangostin group , and LPS/ATP + 40 μmol/L α-mangostin group , respectively) . ELISA experiments were conducted to detect the levels of pro-inflammatory cytokines inter- leukin -6/1β/18 (IL-6 , IL-1β, IL-18) and tumor necrosis factor (TNF-α) in the supernatants of each group , and Western blot was used to detect the expression of NLRP3 , ASC , cleaved caspase-1 , IL-1β, and the phosphoryla- tion levels of p65 (p-p65/p65) in the NF- κB pathway , as well as the expression of p65 in the nuclei of BV-2 cells .
Results :
Compared with the Ctrl group, cell proliferation vitality in the LPS/ATP group was significantly reduced (P < 0. 05) , but low concentrations (10 , 20 , 40 μmol/L) of α-mangostin significantly improved the inhibi- tory effect of LPS/ATP on microglial cell proliferation vitality (P < 0. 05) , while a high concentration (80 μmol/ L) of α-mangostin exacerbated the damage to microglial cells caused by LPS/ATP (P < 0. 05) . C ompared with the Ctrl group , the levels of inflammatory factors IL-6 , IL-1β, IL-18 , TNF-α, and the expression of NLRP3 , ASC , cleaved caspase-1 , IL-1β, and the p-p65/p65 ratio in the 40 μmol/L α-mangostin group , as well as the expression of p65 protein in the nuclei , showed no significant changes ( P > 0 . 05) , whereas these significantly increased in the LPS/ATP group (P < 0. 05) . Compared with the LPS/ATP group , the levels of IL-6 , IL-1β, IL-18 , TNF-α, and the expression of NLRP3 , ASC , cleaved caspase-1 , IL-1β, and the p-p65/p65 ratio in the intervention groups , as well as the expression of p65 protein in the nuclei , decreased in a concentration-dependent manner with increasing α-mangostin concentration , with the most significant reduction ob served in the LPS/ATP + 40 μmol/L α- mangostin group (P < 0. 01) .
Conclusion
α-mangostin can inhibit the neuroinflammatory response mediated by NLRP3 inflammasome activation in BV-2 cells through the NF- κB pathway .
3.Influence of infection frequency and vaccination on virus mutation of SARS-CoV-2
Guo XU ; Huan FAN ; Jianguang FU ; Huiyan YU ; Fei DENG ; Zhuhan DONG ; Shihan ZHANG ; Fengcai ZHU ; Changjun BAO ; Liguo ZHU
Chinese Journal of Experimental and Clinical Virology 2024;38(5):481-488
Objective:To analyze the effects of SARS-CoV-2 infection and vaccination on virus mutation.Methods:The whole genome sequencing sequences of 2 659 local SARS-CoV-2 specimens from Jiangsu Province in 2023 were selected for analysis, and relevant information such as demographic and clinical characteristics were collected, and the effects of infection and vaccination on the genome-wide mutation rate and S gene′s selective pressure of the virus were analyzed by univariate and multivariate linear regression models.Results:The average age of these infected patients was 55.0 (31.0, 74.0) years, 1 150 cases (43.2%) in the age group of ≥60 years, 1 367 cases (51.4%) were males, 2 044 cases (76.9%) had a history of COVID-19 vaccination, and 1 629 cases (61.3%) had the first-time infection. The clinical symptoms of the infected patients were mainly mild, with a total of 2434 cases (91.5%), and 29 cases (1.1%) with severe symptoms or more. The average substitution rate of SARS-CoV-2 was 9.69 (9.38, 9.98)×10 -4 subs/site/year, and the dN/dS value of the S gene was 6.08 (5.56, 8.66), which was significantly greater than that of 1 ( P<0.001), indicating positive selection. The result of univariate and multivariate linear regression model analysis showed that the SARS-CoV-2 substitution rate was higher in those with vaccination history and reinfection, aged 20-30 years, ≥60 years, and the SARS-CoV-2 substitution rate was lower in males with moderate clinical symptoms and severe disease and above. Those with a history of vaccination and reinfection, aged 50-60 years old, ≥60 years old have smaller S gene dN/dS. Conclusions:Under the immune pressure exerted by vaccination and infection, the genome-wide mutation of SARS-COV-2 accelerated, but the non-synonymous mutation rate of the S gene decreased. The mechanism causing these phenomena needs further study.
4.Arterial stiffness in subclinical atherosclerosis quantified with ultrafast pulse wave velocity measurements: a comparison with a healthy population using propensity score matching
Xuezhong JIANG ; Weiming GE ; Hui HUANG ; Yating LI ; Xiaojing LIU ; Huiyan PANG ; Rui HE ; Hui WANG ; Zhengqiu ZHU ; Ping HE ; Yinping WANG ; Xuehui MA ; Airong REN ; Bixiao SHEN ; Meijuan WANG
Ultrasonography 2024;43(4):263-271
Purpose:
This study aimed to evaluate changes in ultrafast pulse wave velocity (ufPWV) in individuals with arterial stiffness and subclinical atherosclerosis (subAS), and to provide cutoff values.
Methods:
This retrospective study recruited 231 participants, including 67 patients with subAS. The pulse wave velocity was measured at the beginning and end of systole (PWV-BS and PWVES, respectively) using ultrafast ultrasonography to assess arterial stiffness. The right and left common carotid arteries were measured separately, and laboratory metabolic parameters were also collected. Participants were balanced between groups using propensity score matching (PSM) at a 1:1 ratio, adjusting for age, sex, and waist-to-hip ratio as potential confounders. Cutoff values of ufPWV for monitoring subAS were determined via receiver operating characteristic (ROC) curve analysis.
Results:
PWV-ES, unlike PWV-BS, was higher in the subAS subgroup than in the subAS-free group after PSM (all P<0.05). For each 1 m/s increase in left, right, and bilateral mean PWV-ES, the risk of subAS increased by 23% (95% confidence interval [CI], 1.04 to 1.46), 26% (95% CI, 1.07 to 1.52), and 38% (95% CI, 1.12 to 1.72), respectively. According to ROC analyses, predictive potential was found for left PWV-ES (cutoff value=7.910 m/s, P=0.002), right PWV-ES (cutoff value=6.615 m/s, P=0.003), and bilateral mean PWV-ES (cutoff value=7.415 m/s, P<0.001), but not for PWV-BS (all P>0.05).
Conclusion
PWV-ES measured using ultrafast ultrasonography was significantly higher in individuals with subAS than in those without. Specific PWV-ES cutoff values showed potential for predicting an increased risk of subAS.
5.Establishing integrated system-based point-of-care testing method for genotyping of respiratory syncytial virus
Fei DENG ; Liguo ZHU ; Ke XU ; Xian QI ; Huiyan YU ; Shenjiao WANG ; Changjun BAO
Chinese Journal of Microbiology and Immunology 2024;44(8):660-664
Objective:To develop an integrated point-of-care testing (POCT) reagent for genotying respiratory syncytial virus (RSV) and evaluate its performance.Methods:Specific primers and probes were designed based on the conserved sequences of the genomes of RSV A and B as well as ON1 and BA9 genotypes. The PCR reaction system and conditions were optimized. The vitrification technology of reagents and multiplex detection platform were integrated to develop the RSV genotyping POCT reagent. The sensitivity, specificity, reproducibility, and clinical performance of the product were then evaluated.Results:The sensitivity of the developed integrated RSV genotyping POCT reagent reached 500 copies/ml. It exhibited good specificity with no cross-reaction with clinically similar pathogens. The coefficient of variation of Ct values for both inter-batch and intra-batch reproducibility was less than 5%, indicating good reproducibility. In testing 53 clinical samples, the detection results showed high consistency and concordance with the reference reagent, with a positive concordance rate of up to 98.11%.Conclusions:The developed integrated RSV genotyping POCT reagent incorporates nucleic acid extraction, purification, and detection into a single process, achieving a "sample in, result out" workflow. It is simple to operate and provides accurate, reliable, and stable detection results. This product can be used for the genotyping of RSV A and B in POCT, offering support for the prevention, control, and diagnosis of RSV.
6.Arterial stiffness in subclinical atherosclerosis quantified with ultrafast pulse wave velocity measurements: a comparison with a healthy population using propensity score matching
Xuezhong JIANG ; Weiming GE ; Hui HUANG ; Yating LI ; Xiaojing LIU ; Huiyan PANG ; Rui HE ; Hui WANG ; Zhengqiu ZHU ; Ping HE ; Yinping WANG ; Xuehui MA ; Airong REN ; Bixiao SHEN ; Meijuan WANG
Ultrasonography 2024;43(4):263-271
Purpose:
This study aimed to evaluate changes in ultrafast pulse wave velocity (ufPWV) in individuals with arterial stiffness and subclinical atherosclerosis (subAS), and to provide cutoff values.
Methods:
This retrospective study recruited 231 participants, including 67 patients with subAS. The pulse wave velocity was measured at the beginning and end of systole (PWV-BS and PWVES, respectively) using ultrafast ultrasonography to assess arterial stiffness. The right and left common carotid arteries were measured separately, and laboratory metabolic parameters were also collected. Participants were balanced between groups using propensity score matching (PSM) at a 1:1 ratio, adjusting for age, sex, and waist-to-hip ratio as potential confounders. Cutoff values of ufPWV for monitoring subAS were determined via receiver operating characteristic (ROC) curve analysis.
Results:
PWV-ES, unlike PWV-BS, was higher in the subAS subgroup than in the subAS-free group after PSM (all P<0.05). For each 1 m/s increase in left, right, and bilateral mean PWV-ES, the risk of subAS increased by 23% (95% confidence interval [CI], 1.04 to 1.46), 26% (95% CI, 1.07 to 1.52), and 38% (95% CI, 1.12 to 1.72), respectively. According to ROC analyses, predictive potential was found for left PWV-ES (cutoff value=7.910 m/s, P=0.002), right PWV-ES (cutoff value=6.615 m/s, P=0.003), and bilateral mean PWV-ES (cutoff value=7.415 m/s, P<0.001), but not for PWV-BS (all P>0.05).
Conclusion
PWV-ES measured using ultrafast ultrasonography was significantly higher in individuals with subAS than in those without. Specific PWV-ES cutoff values showed potential for predicting an increased risk of subAS.
7.Arterial stiffness in subclinical atherosclerosis quantified with ultrafast pulse wave velocity measurements: a comparison with a healthy population using propensity score matching
Xuezhong JIANG ; Weiming GE ; Hui HUANG ; Yating LI ; Xiaojing LIU ; Huiyan PANG ; Rui HE ; Hui WANG ; Zhengqiu ZHU ; Ping HE ; Yinping WANG ; Xuehui MA ; Airong REN ; Bixiao SHEN ; Meijuan WANG
Ultrasonography 2024;43(4):263-271
Purpose:
This study aimed to evaluate changes in ultrafast pulse wave velocity (ufPWV) in individuals with arterial stiffness and subclinical atherosclerosis (subAS), and to provide cutoff values.
Methods:
This retrospective study recruited 231 participants, including 67 patients with subAS. The pulse wave velocity was measured at the beginning and end of systole (PWV-BS and PWVES, respectively) using ultrafast ultrasonography to assess arterial stiffness. The right and left common carotid arteries were measured separately, and laboratory metabolic parameters were also collected. Participants were balanced between groups using propensity score matching (PSM) at a 1:1 ratio, adjusting for age, sex, and waist-to-hip ratio as potential confounders. Cutoff values of ufPWV for monitoring subAS were determined via receiver operating characteristic (ROC) curve analysis.
Results:
PWV-ES, unlike PWV-BS, was higher in the subAS subgroup than in the subAS-free group after PSM (all P<0.05). For each 1 m/s increase in left, right, and bilateral mean PWV-ES, the risk of subAS increased by 23% (95% confidence interval [CI], 1.04 to 1.46), 26% (95% CI, 1.07 to 1.52), and 38% (95% CI, 1.12 to 1.72), respectively. According to ROC analyses, predictive potential was found for left PWV-ES (cutoff value=7.910 m/s, P=0.002), right PWV-ES (cutoff value=6.615 m/s, P=0.003), and bilateral mean PWV-ES (cutoff value=7.415 m/s, P<0.001), but not for PWV-BS (all P>0.05).
Conclusion
PWV-ES measured using ultrafast ultrasonography was significantly higher in individuals with subAS than in those without. Specific PWV-ES cutoff values showed potential for predicting an increased risk of subAS.
8.Arterial stiffness in subclinical atherosclerosis quantified with ultrafast pulse wave velocity measurements: a comparison with a healthy population using propensity score matching
Xuezhong JIANG ; Weiming GE ; Hui HUANG ; Yating LI ; Xiaojing LIU ; Huiyan PANG ; Rui HE ; Hui WANG ; Zhengqiu ZHU ; Ping HE ; Yinping WANG ; Xuehui MA ; Airong REN ; Bixiao SHEN ; Meijuan WANG
Ultrasonography 2024;43(4):263-271
Purpose:
This study aimed to evaluate changes in ultrafast pulse wave velocity (ufPWV) in individuals with arterial stiffness and subclinical atherosclerosis (subAS), and to provide cutoff values.
Methods:
This retrospective study recruited 231 participants, including 67 patients with subAS. The pulse wave velocity was measured at the beginning and end of systole (PWV-BS and PWVES, respectively) using ultrafast ultrasonography to assess arterial stiffness. The right and left common carotid arteries were measured separately, and laboratory metabolic parameters were also collected. Participants were balanced between groups using propensity score matching (PSM) at a 1:1 ratio, adjusting for age, sex, and waist-to-hip ratio as potential confounders. Cutoff values of ufPWV for monitoring subAS were determined via receiver operating characteristic (ROC) curve analysis.
Results:
PWV-ES, unlike PWV-BS, was higher in the subAS subgroup than in the subAS-free group after PSM (all P<0.05). For each 1 m/s increase in left, right, and bilateral mean PWV-ES, the risk of subAS increased by 23% (95% confidence interval [CI], 1.04 to 1.46), 26% (95% CI, 1.07 to 1.52), and 38% (95% CI, 1.12 to 1.72), respectively. According to ROC analyses, predictive potential was found for left PWV-ES (cutoff value=7.910 m/s, P=0.002), right PWV-ES (cutoff value=6.615 m/s, P=0.003), and bilateral mean PWV-ES (cutoff value=7.415 m/s, P<0.001), but not for PWV-BS (all P>0.05).
Conclusion
PWV-ES measured using ultrafast ultrasonography was significantly higher in individuals with subAS than in those without. Specific PWV-ES cutoff values showed potential for predicting an increased risk of subAS.
9.Arterial stiffness in subclinical atherosclerosis quantified with ultrafast pulse wave velocity measurements: a comparison with a healthy population using propensity score matching
Xuezhong JIANG ; Weiming GE ; Hui HUANG ; Yating LI ; Xiaojing LIU ; Huiyan PANG ; Rui HE ; Hui WANG ; Zhengqiu ZHU ; Ping HE ; Yinping WANG ; Xuehui MA ; Airong REN ; Bixiao SHEN ; Meijuan WANG
Ultrasonography 2024;43(4):263-271
Purpose:
This study aimed to evaluate changes in ultrafast pulse wave velocity (ufPWV) in individuals with arterial stiffness and subclinical atherosclerosis (subAS), and to provide cutoff values.
Methods:
This retrospective study recruited 231 participants, including 67 patients with subAS. The pulse wave velocity was measured at the beginning and end of systole (PWV-BS and PWVES, respectively) using ultrafast ultrasonography to assess arterial stiffness. The right and left common carotid arteries were measured separately, and laboratory metabolic parameters were also collected. Participants were balanced between groups using propensity score matching (PSM) at a 1:1 ratio, adjusting for age, sex, and waist-to-hip ratio as potential confounders. Cutoff values of ufPWV for monitoring subAS were determined via receiver operating characteristic (ROC) curve analysis.
Results:
PWV-ES, unlike PWV-BS, was higher in the subAS subgroup than in the subAS-free group after PSM (all P<0.05). For each 1 m/s increase in left, right, and bilateral mean PWV-ES, the risk of subAS increased by 23% (95% confidence interval [CI], 1.04 to 1.46), 26% (95% CI, 1.07 to 1.52), and 38% (95% CI, 1.12 to 1.72), respectively. According to ROC analyses, predictive potential was found for left PWV-ES (cutoff value=7.910 m/s, P=0.002), right PWV-ES (cutoff value=6.615 m/s, P=0.003), and bilateral mean PWV-ES (cutoff value=7.415 m/s, P<0.001), but not for PWV-BS (all P>0.05).
Conclusion
PWV-ES measured using ultrafast ultrasonography was significantly higher in individuals with subAS than in those without. Specific PWV-ES cutoff values showed potential for predicting an increased risk of subAS.
10.Photohardening therapy for idiopathic actinic dermatoses
Huiyan DENG ; Jiaoquan CHEN ; Huaping LI ; Quan CHEN ; Sanquan ZHANG ; Huilan ZHU
Chinese Journal of Dermatology 2023;56(12):1091-1095
Photohardening therapy, also known as photodesensitization therapy, refers to the phototherapy and photochemotherapy of idiopathic actinic dermatoses, and its goal is to improve the patients′ tolerance to sunlight and prevent disease flares. Its mechanisms of action involve a variety of cellular and inflammatory factors. This therapy is suitable for all idiopathic actinic dermatoses, with definite efficacy and good safety. However, the treatment specificity usually leads to poor compliance. The development of UVA1 rush hardening and home phototherapy is expected to solve this problem.


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