1.Analysis of Mechanism of Astragaloside Ⅳ in Regulating Ferroptosis Through SLC7A11/GPX4 Pathway Against Vascular Smooth Muscle Cell Proliferation
Guoting LI ; Changchao YANG ; Lin LIU ; Weikang LI ; Zixian ZHAO ; Quan SHEN ; Jingshan ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):159-167
ObjectiveTo investigate the effect of astragaloside Ⅳ(AS-Ⅳ) on the proliferation of vascular smooth muscle cells(VSMCs) induced by angiotensin Ⅱ(Ang Ⅱ) based on solute carrier family 7 member 11/glutathione peroxidase 4(SLC7A11/GPX4) pathway. MethodsPrimary rat thoracic aortic VSMCs were cultured by tissue explant method, and the cell types were identified by immunofluorescence. Cell counting kit-8(CCK-8) was used to determine the optimal concentration and time of AS-Ⅳ after Ang Ⅱ stimulation. The experiment was divided into blank group, model group, AS-Ⅳ group(40 μmol·L-1), Erastin group(0.5 μmol·L-1), Erastin+AS-Ⅳ group(0.5 μmol·L-1+40 μmol·L-1). The blank group was cultured in normal medium, the model group was cultured in medium containing Ang Ⅱ(0.1 μmol·L-1), and each administration group was cultured in medium containing Ang Ⅱ(0.1 μmol·L-1) and the corresponding doses of drug. CCK-8 and plate clone formation assay were used to detect the proliferation of cells in each group, Prussian blue staining was used to detect cell iron deposition, the content of reactive oxygen species(ROS) in cells was detected by fluorescence probe method, the content of malondialdehyde(MDA) was detected by thiobarbituric acid(TBA) method, and the protein levels of SLC7A11 and GPX4 in each group were detected by Western blot. ResultsPrimary rat thoracic aortic VSMCs were successfully cultured by tissue explant method, and immunofluorescence detection showed that positive expression of α-smooth muscle actin(α-SMA) and negative expression of vimentin in the cells, identifying them as VSMCs. The optimal concentration and time of AS-Ⅳ determined by CCK-8 were 40 μmol·L-1 and 24 h, respectively. Pharmacodynamic studies showed that compared with the blank group, the cell proliferation in the model group increased, the iron deposition in the cells increased, the contents of ROS and MDA increased, and the expression levels of SLC7A11 and GPX4 proteins decreased(P<0.05, P<0.01). Compared with the model group, the cell proliferation of the AS-Ⅳ group was inhibited, the iron deposition in the cells was decreased, the contents of ROS and MDA were decreased, and the expression levels of SLC7A11 and GPX4 proteins were increased(P<0.05, P<0.01). While in the Erastin group, the cell proliferation was increased, the iron deposition was increased, ROS and MDA contents were increased, and the expression levels of SLC7A11 and GPX4 proteins were decreased(P<0.05, P<0.01). Compared with the AS-Ⅳ group, Erastin+AS-Ⅳ group showed increased cell proliferation, increased iron deposition in cells, increased ROS and MDA contents, and decreased expression of SLC7A11 and GPX4 proteins(P<0.05). Compared with the Erastin group, the cell proliferation in Erastin+AS-Ⅳ group was inhibited, the iron deposition was decreased, the contents of ROS and MDA were decreased, and the expression levels of SLC7A11 and GPX4 proteins were increased(P<0.05, P<0.01). ConclusionAS-Ⅳ can inhibit ferroptosis by regulating the SLC7A11/GPX4 pathway, so as to weaken the proliferation of VSMCs, thus playing a role in the treatment of atherosclerosis.
2.Mechanism of Modified Tianwang Buxindan on Skin of Sleep-deprived Mice Through PI3K/Akt/Nrf2 Signaling Pathway
Juanping CHEN ; Yuan PENG ; Xuemin HONG ; Li YANG ; Bo XU ; Chong ZHANG ; Xuelin GUO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):120-128
ObjectiveTo observe the effect of modified Tianwang Buxindan (MTBD) on the skin of sleep-deprived (SD) mice and investigate its mechanism. MethodSixty 2-month-old female Kunming mice were randomly divided into a blank group, a model group, a vitamin C (VC, 0.08 g·kg-1), and MTBD low-, medium-, and high-dose groups (6.5, 12.5, 25 g·kg-1). Except for the blank group, the other groups were subjected to SD mouse model induction (using multiple platform water environment method for 18 hours of sleep deprivation daily from 15:00 to next day 9:00), continuously for 14 days, and caffeine (CAF, 7.5 mg·kg-1) was injected intraperitoneally from the 2nd week onwards, continuously for 7 days. While modeling, the blank group and the model group were administered with normal saline (0.01 mL·g-1), and the other groups received corresponding drugs for treatment. On the day of the experiment, general observations were recorded (such as body weight, spirit, fur, and skin). After sampling, skin tissue pathological changes were observed under an optical microscope using hematoxylin-eosin (HE) and Masson staining methods. Skin thickness and skin moisture content were measured. Biochemical assay kits were used to detect skin hydroxyproline (HYP) content, skin and serum superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-1β levels in mice. Western blot was used to detect skin tissue type Ⅰ collagen (ColⅠ), type Ⅲ collagen (ColⅢ), phosphatidylinositol 3-kinase (PI3K), phosphorylated (p)-PI3K, protein kinase B (Akt), p-Akt, nuclear factor E2-related factor 2 (Nrf2), heme oxygenase (HO)-1, and nuclear factor (NF)-κB protein expression. ResultCompared with the blank group, the model group showed varying degrees of changes. In general, signs of aging such as reduced body weight (P<0.01), listlessness, dull fur color, and formation of wrinkles on the skin appeared. Tissue specimen testing revealed skin thinning, flattening of the dermoepidermal junction (DEJ), and reduced collagen fibers under the optical microscope. Skin thickness and moisture content decreased, skin tissue HYP content significantly decreased (P<0.01), skin and serum SOD activity significantly decreased (P<0.01), and MDA content significantly increased (P<0.01). Serum IL-6, TNF-α, and IL-1β levels significantly increased (P<0.01). Skin ColⅠ, ColⅢ, p-PI3K/PI3K, p-Akt/Akt, Nrf2, and HO-1 protein expression significantly decreased (P<0.05, P<0.01), and NF-κB expression increased (P<0.01). Compared with the model group, the VC group and the MTBD low-dose group showed increased skin moisture content, HYP content, SOD activity, and ColⅠ, ColⅢ, p-PI3K/PI3K protein expression (P<0.05, P<0.01), and decreased serum MDA content (P<0.05). In addition, a decrease in serum IL-6 and IL-1β levels was detected in the MTBD low-dose group (P<0.05), while the above indicators in the MTBD medium- and high-dose groups improved (P<0.05, P<0.01). ConclusionSleep deprivation accelerates the aging process of the skin in SD model mice. MTBD can improve this phenomenon, exerting anti-inflammatory and antioxidant effects, and its mechanism of action may be related to the activation of the PI3K/Akt/Nrf2 signaling pathway.
3.Clinical blood dispatching between hospitals in Shijiazhuang
Wentao XIE ; Yintu MA ; Li CHEN ; Jia LIU ; Jingshan LIU ; Lihua LI
Chinese Journal of Blood Transfusion 2023;36(11):1015-1018
【Objective】 To analyze the blood dispatching of hospitals in Shijiazhuang area, in order to provide reference for blood dispatching policy of hospitals and ensure clinical blood supply. 【Methods】 The hospital blood dispatching data from 2018 to 2021 were queried through the blood center information management system (SHINOW9.5) and TMIS system, and the blood dispatching volume, frequency, variety and blood type distribution of hospitals at all levels were analyzed by SPSS statistical software. 【Results】 From 2018 to 2021, there were 1 196 times of blood dispatching in hospitals with a total volume of 4 648.5 U, showing a downward and then upward trend two years before and after the outbreak of COVID-19. As the main blood dispatching product, erythrocyte dispatching was 3 988 U, accounting for 85.79% of the total dispatching volume, with a dispatching rate of 2.99‰. The dispatching volume of red blood cells in secondary hospitals accounted for 86.13%.The dispatching rate of erythrocyte in secondary and tertiary hospitals was 1.03% and 0.06%, respectively, with significant difference(P<0.05). The dispatching rates of type A and AB red blood cells were 3.08‰ and 4.97‰, which were significantly different from those of type B and O red blood cells. 【Conclusion】 Blood dispatching has become an effective way to avoid blood waste and ensure emergency blood use in secondary and tertiary hospitals. In the actual operation, health administration department should strengthen supervision and hospitals should improve the blood dispatching process to ensure blood safety.
4.Path analysis of quality of life in stroke patients based on structural equation model
Shiheng TANG ; Jingshan LIN ; Yiyang WANG ; Xiaoxuan QIAO ; Yuxia LI
Chinese Journal of Nursing 2023;58(22):2747-2753
Objective To construct a structural equation model to analyze the effect of various factors on the quality of life in stroke patients.Methods In this cross-sectional study,a total of 657 stroke patients from 2 ter-tiary A hospitals in Shanghai were enrolled as investigation subjects via convenience sampling methodology.General information questionnaire,Sense of Coherence Scale-13,Perceived Social Support Scale,Medical Coping Modes Questionnaire,National Institutes of Health Stroke Scale,Self-Esteem Scale and Stroke-Specific Quality of Life were utilized as the investigating tools;Mplus 7.4 was used to construct a structural equation model.Results The aver age score of quality of life in stroke patients was(178.30±30.30);neurological status,self-esteem were negatively cor-related with quality of life(P<0.05);sense of coherence,social support,coping style were positively correlated with quality of life(P<0.05);sense of coherence,taking a confrontational approach for coping,yielding and neurological status had a direct impact on quality of life(path coefficients were 0.471,0.169、0.135,-0.089,P<0.05);social sup-port,self-esteem,sense of coherence and neurological status had an indirect impact on quality of life(path coefficients were 0.232,0.108,0.058,-0.058,P<0.001).Conclusion The quality of life in stroke patients is influenced by multi-ple factors interactively,and sense of coherence is an important factor affecting the quality of life;adopting a confron-tational approach as coping styles,good social support and high self-esteem can improve the level of quality of life.
5.Value of soluble triggering receptor expression on myeloid cells-1 level of alveolar fluid in early diagnosis of ventilator-associated pneumonia: a Meta-analysis
Jiayu TAN ; Xiang HUANG ; Minzhi WU ; Bo YANG ; Zhengchao LUO ; Jingshan LI ; Yuanfang ZHONG ; Zhenpeng LIANG
Chinese Critical Care Medicine 2020;32(9):1067-1073
Objective:Serum soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) is a useful biomarker of bacterial infection. However, the diagnostic value of sTREM-1 of alveolar fluid in pulmonary infection is still unclear. This article aimed to explore the value of sTREM-1 of alveolar fluid in the early diagnosis of ventilator-associated pneumonia (VAP) by systematic review of relevant literatures.Methods:CNKI, Wanfang, VIP, PubMed/Medline and Embase databases were retrieved. Articles on diagnosis of VAP by sTREM-1 before June 30, 2019 were collected. QUADAS-2 scale provided by Cochrane Collaboration Network was used to evaluate the quality of diagnostic experiments. RevMan 5.3 and Stata 13.0 software were used to complete Meta-analysis. The levels of sTREM-1 between VAP and non-VAP patients were analyzed by Meta-analysis, and then diagnostic test Meta-analysis was conducted. Heterogeneity, sensitivity and publication bias were analyzed.Results:A total of 24 articles were enrolled. QUADAS-2 scale indicated that the selected literature had low bias and high clinical adaptability. ① In Meta-analysis of sTREM-1 level in alveolar fluid, 20 articles were selected and found to have high heterogeneity ( I2 = 94.4%, P = 0.000). The random effects models were used for Meta-analysis. It was indicated that the sTREM-1 level in alveolar fluid of VAP group was significantly higher than that of non-VAP group with significant difference [standardized mean difference ( SMD) was 1.47, 95% confidence interval (95% CI) was 1.00-1.95, Z = 6.14, P = 0.000]. By subgroup analysis and Meta-regression analysis, no source of heterogeneity was found. Sensitivity analysis suggested that the results of this Meta-analysis were robust and credible, and Begg funnel plot analysis showed that there was no significant publication bias ( Z = 1.46, P = 0.143). ② A total of 18 articles were included in the Meta-analysis of diagnostic experiments. Deek funnel plot showed publication bias ( P = 0.012). The combined sensitivity was 0.87 (95% CI was 0.81-0.91), specificity was 0.80 (95% CI was 0.73-0.86), and diagnostic odds ratio ( DOR) was 26 (95% CI was 13-50). Subgroup analysis of three different sources of alveolar fluid (bronchoalveolar lavage fluid, endotracheal aspiration fluid and exhaled ventilator condensate) showed that STREM-1 had a certain value in early diagnosis of VAP. The I2 of combined DOR was 35.4%, and I2 of sensitivity was 79.46%, I2 of specificity was 77.61%, suggesting heterogeneity in the selected literature. Subgroup analysis found that nationality, subject design, sample source, sample size and diagnostic "gold criteria" were related to heterogeneity, but not age. The area under synthetic receiver operating characteristic (SROC) curve (AUC) was 0.90 (95% CI was 0.87-0.92). Conclusions:The detection of sTREM-1 level in alveolar fluid can be used for the early diagnosis of VAP with high sensitivity and specificity. If combined with other biomarkers, it may have more diagnostic value.
6.Directional breeding of high oil content peanut variety Yuhua 9 by in vitro mutagenesis and screening.
Jingshan WANG ; Yanan JIANG ; Xiubo YIN ; Yanjun YI ; Jian ZHAO ; Puxiang SHI ; Songjian LI ; Shanlin YU
Chinese Journal of Biotechnology 2019;35(7):1277-1285
Leaf water potential of peanut subjected to drought stress is positively related to the oil content of peanut kernels. The aim of this study was to directly screen the high oil mutants of peanut and create the new peanut varieties using hydroxyproline as water potential regulator. In vitro mutagenesis was carried out with the embryonic leaflets of peanut variety Huayu 20 as explants and pingyangmycin as a mutagen added into the somatic embryo formation medium. The formed somatic embryos were successively transferred to somatic embryo germination and selection medium containing 6 mmol/L hydroxyproline (at -2.079 MPa water potential ) to induce regeneration and directionally screen high oil content mutants. After that, these plantlets were grafted and transplanted to the experimental field and 132 high oil mutants with oil content over 55% were obtained from the offspring of regenerated plants. Finally, among them, the oil contents of 27 lines were higher than 58% and of 2 lines were higher than 60%. A new peanut variety Yuhua 9 with high yield and oil content was bred from the regenerated plant progenies combining the pedigree breeding method. The yield was 14.0% higher than that of the control cultivar in the testing new peanut varieties of Liaoning province, and also it has passed the national registration of non-major crop varieties. Yuhua 9 with an oil content of 61.05%, which was 11.55 percentage points higher than that of the parent Huayu 20, was the peanut cultivar with the highest oil content in the world. The result showed that it was an effective way for directional breeding of high oil peanut varieties by means of the three-step technique including in vitro mutagenesis, directional screening by reducing water potential in medium and pedigree selection of regenerated plant progenies.
Arachis
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Droughts
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Germination
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Mutagenesis
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Plant Breeding
7.Breeding peanut variety Yuhua 7 by fast neutron irradiation and tissue culture.
Xia WANG ; Luxiang LIU ; Lixian QIAO ; Jiongming SUI ; Defeng JIANG ; Guan LI ; Linshu ZHAO ; Jingshan WANG
Chinese Journal of Biotechnology 2019;35(2):270-280
Creating new germplasms and breeding new cultivars in peanut by radiation mutagenesis and tissue culture were conducted in this study, aiming to develop new breeding method of peanut. Mature seeds from Luhua 11, the most commonly grown peanut cultivar in Northern China, were treated by fast neutron irradiation. Then the embryo leaflets were separated from the irradiated seeds and inoculated on the media, and the regenerated seedlings were obtained through somatic embryogenesis pathway. The regenerated seedlings were grafted, acclimated and then transplanted into field and the selfed pods were harvested from 83 regenerated plants. The progenies were selected by the pedigree method, and 107 mutants were obtained from the progenies of the 83 regenerated plants. Different mutants showed obvious variation in many agronomic traits, including main stem height, branch number, pod shape and size, seed coat color, inner seed coat color, oil content and protein content etc. Yuhua 7, a new peanut variety with low oil content, early maturity and waterlogging tolerance was obtained. The yield of Yuhua 7 was over 14% higher than that of the mutagenic parent Luhua 11, and the oil content of kernels was 47.0%, lower than that of parent Luhua 11 with 52.1% oil content. Yuhua 7 had passed peanut variety regional multi-location trials in Liaoning Province in 2016 and its average yield was 13.8% higher than that of the control variety Baisha 1017. It had also passed national peanut variety registration, and the registration ID is "GPD peanut (2018) 370105". The results show that irradiation mutagenesis combined with tissue culture is an effective method for creating new germplasm and breeding new varieties of peanut.
Arachis
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Breeding
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China
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Fast Neutrons
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Plant Breeding
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Seeds
8.Combination of berberine and ciprofloxacin reduces multi-resistant Salmonella strain biofilm formation by depressing mRNA expressions of luxS, rpoE, and ompR
Chenxi SHI ; Minmin LI ; Ishfaq MUHAMMAD ; Xin MA ; Yicong CHANG ; Rui LI ; Changwen LI ; Jingshan HE ; Fangping LIU
Journal of Veterinary Science 2018;19(6):808-816
Bacterial biofilms have been demonstrated to be closely related to clinical infections and contribute to drug resistance. Berberine, which is the main component of Coptis chinensis, has been reported to have efficient antibacterial activity. This study aimed to investigate the potential effect of a combination of berberine with ciprofloxacin (CIP) to inhibit Salmonella biofilm formation and its effect on expressions of related genes (rpoE, luxS, and ompR). The fractional inhibitory concentration (FIC) index of the combination of berberine with CIP is 0.75 showing a synergistic antibacterial effect. The biofilm's adhesion rate and growth curve showed that the multi-resistant Salmonella strain had the potential to form a biofilm relative to that of strain CVCC528, and the antibiofilm effects were in a dose-dependent manner. Biofilm microstructures were rarely observed at 1/2 × MIC/FIC concentrations (MIC, minimal inhibition concentration), and the combination had a stronger antibiofilm effect than each of the antimicrobial agents used alone at 1/4 × FIC concentration. LuxS, rpoE, and ompR mRNA expressions were significantly repressed (p < 0.01) at 1/2 × MIC/FIC concentrations, and the berberine and CIP combination repressed mRNA expressions more strongly at the 1/4 × FIC concentration. The results indicate that the combination of berberine and CIP has a synergistic effect and is effective in inhibiting Salmonella biofilm formation via repression of luxS, rpoE, and ompR mRNA expressions.
Anti-Infective Agents
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Berberine
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Biofilms
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Ciprofloxacin
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Coptis
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Drug Combinations
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Drug Resistance
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Drug Resistance, Multiple
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Repression, Psychology
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RNA, Messenger
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Salmonella
9.Breeding of peanut variety Yuhua 4 by in vitro mutagenesis.
Guan LI ; Xia WANG ; Xiubo YIN ; Xiaohui HU ; Jing CHEN ; Lixian QIAO ; Jiongming SUI ; Jingshan WANG
Chinese Journal of Biotechnology 2017;33(5):766-774
The embryonic leaflets of peanut (Arachis hypogaea) variety Huayu 20 were used as explants and pingyangmycin as a mutagen to induce somatic embryos. Four weeks after the inoculation, the survived explants were transferred to somatic embryo germination medium containing screening reagent hydroxyproline, and finally 15 regenerated plants were obtained. Pedigree breeding method was used during the following selection breeding, and three lines with significantly increased yield and 23 lines with high oil content were obtained from these mutant offsprings. The line with both high yield and high oil content has passed peanut variety multi-location in Anhui province and was named "Yuhua 4". Its yield was 16.63% higher than that of the control variety Baisha 1016, ranking the first in all the testing varieties. Yuhua 4 showed the characteristics of early maturity, small pod and high oil content. The oil content of kernels was 56.10%, higher than that of original parent Huayu 20 with 49.50% oil content, tested by the Ministry of Agriculture of Oil and Products Quality Supervision, Inspection and Test Center (Wuhan), and the yield was 15% higher than that of Huayu 20. It was concluded that in vitro mutagenesis and target screening was an effective way on creating new germplasm and breeding new variety in peanut.
10.Roscovitine inhibits inflammatory hyperplasia of carotid artery intima in rats via suppressing nuclear factor-κB activation
Jingshan ZHAO ; Jiahuan SUN ; Kun YU ; Yu LIU ; Chao WANG ; Aiying LI
Chinese Journal of Pathophysiology 2017;33(2):233-238
AIM:To study the effect of roscovitine on the inflammatory hyperplasia of carotid artery intima in rats and the related mechanisms .METHODS: SD rats ( n=60 ) were randomly divided into 3 groups including control group, model group and treatment group .The rat model was established by trypsin digestion injury .The rats in control group were given sham operation .The rats in treatment group were administered with 0.5 mL roscovitine (2 g/L) slow-re-leasing gelatin.The rats in each group were fed normally for 4 weeks, then killed to take out carotid arteries for further ob-servations .The effects of roscovitine on the inflammatory hyperplasia of carotid artery intima and the related mechanism via nuclear factor-κB ( NF-κB) in the rats were detected by Western blot .RESUITS:Roscovitine inhibited the activation of NF-κB and the expression of inflammatory factors cyclooxygenase-2 (COX-2), vascular cell adhesion molecule-1 (VCAM-1),TNF-αand IL-6 via blocking the phosphorylation activation of NF-κB and inhibiting the degradation of IκB-α.CON-CLUSION:Roscovitine inhibits inflammatory hyperplasia of carotid artery intima in the rats via suppressing NF-κB activa-tion.

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