1.Effect of aconite decoction on the activity and polarization of mouse RAW264.7 macrophages
Mingcong SHAO ; Hubo CHEN ; Yidan ZHANG ; Ziyan LI ; Lina WANG
Journal of Pharmaceutical Practice and Service 2025;43(7):329-334
Objective To investigate the effects of Aconite decoction (AD) on the viability and polarization of murine RAW264.7 macrophages induced by lipopolysaccharide (LPS) or interleukin-4 (IL-4). Methods Cytotoxicity of AD was assessed by the CCK-8 assay. RAW264.7 cells were polarized toward M1 phenotype by LPS or M2 phenotype by IL-4, followed by treatment with varying concentrations of AD. Macrophage polarization was analyzed by flow cytometry. Quantitative PCR was performed to measure mRNA expression of polarization-associated markers (IL-6, iNOS, Arg1, and Ym1). ELISA was used to quantify secreted cytokines (TNF-α and IL-10)in the supernatant. Results At non-toxic concentrations, IL-6 and iNOS mRNA levels in LPS-stimulated cells were significantly upregulated while Arg1 and Ym1 expression in IL-4-treated groups were downregulated by AD. Concurrently, TNF-α secretion in LPS-induced M1 polarization was enhanced but IL-10 production in IL-4-induced M2 polarization was suppressed by AD. Conclusion AD could promote macrophage proliferation and viability, augments LPS-driven M1 polarization, and inhibit IL-4-mediated M2 polarization, which provided experimental evidence for the potential application of AD in tumor immunotherapy.
2.Effects of laminarin on non-proliferative diabetic retinopathy in C57BL/6 mice based on transcriptomics analysis
Lei Zhang ; Sumei Zhang ; Zhen Yang ; Weikang Hu ; Hongmei Bai ; Wenjing Zhou ; Zihan Wang ; Mingcong Li ; Shengquan Zhang ; Rongfeng Liao
Acta Universitatis Medicinalis Anhui 2025;60(3):392-398
Objective :
To investigate the effect of laminarin(LAM) on nonproliferative diabetes retinopathy by high throughput sequencing(RNA-seq).
Methods :
The diabetes model was established by intraperitoneal injection of streptozotocin(STZ), and the effect of LAM on diabetic mice was observed.C57BL/6 mice were randomly divided into three groups: Control group, Model group, and LAM group, with 8 mice in each group. After 8 weeks of modeling, the LAM group received a 4-week intraperitoneal injection of LAM treatment. Changes in blood glucose and body weight of the three groups of mice were recorded, HE staining was performed to examine retinal lesions, and RNA-seq was used to identify differentially expressed genes(DEGs) in diabetic retinopathy(DR) under the action of STZ and LAM.
Results :
STZ successfully established the model of DR, and LAM reduced the blood sugar in diabetic mice to a certain extent and improved the pathological morphology of retinal structural looseness in diabetic mice. After RNA-seq analysis of DEGs, it was found that there were a total of 214 DEGs in the retina of the Model group mice compared to the Control group. Enrichment analysis revealed that DR could exacerbate the lesions through the PI3K Akt signaling pathway. There were a total of 42 DEGs in the retina of the Model group and LAM group mice, and enrichment showed that LAM improved the lesions through the neutrophil extracellular trap pathway. Early growth response factor 1(Egr1), FBJ osteosarcoma oncogene(Fos), nuclear receptor subfamily 4A member 1(Nr4a1), and salt-induced kinase 1(Sik1) were regulated by STZ, and LAM significantly regulated their expression, which might be closely related to LAM′s treatment of diabetic retinopathy.
Conclusion
DEGs can exacerbate the severity of diabetic retinopathyviathe PI3K-Akt signaling pathway. LAM can mitigate diabetic retinopathyviathe neutrophil extracellular trap pathway. Egr1, Fos, Nr4a1, and Sik1 are key genes involved in LAM treatment of STZ-induced DR.
3.Effects of LSS function deficiency on intestinal function in NAFLD model mice
Hongmei Bai ; Zhen Yang ; Weikang Hu ; Zihan Wang ; Wenjing Zhou ; Qingya He ; Jian Zhong ; Mingcong Li ; Li Liu ; Chaoyang Zhang ; Sumei Zhang ; Shengquan Zhang
Acta Universitatis Medicinalis Anhui 2025;60(9):1653-1660
Objective:
To investigate the effect of loss of function of lanosterol synthase( LSS) gene on intestinal function in a mouse model of non-alcoholic fatty liver disease( NAFLD) induced by a high-fat diet.
Methods:
LSS gene heterozygous knockout C57 mice ( LSS + / -) were established using the CRISRP / Cas9 system.After being fed a high-fat diet with 60% fat content for 6 months,the fat deposition in liver tissues was detected by HE and Oil red O staining,the morphological changes of small intestine tissue were detected by HE staining.The changes in total cholesterol content in intestinal tissue were detected by kits.The gastrointestinal motility function of mice was detected by phenol red paste.The intestinal permeability was detected by Evans blue staining,and the expression of LSS,tight junction protein ( Claudin) -1,Claudin-5,cluster of differentiation 36 ( CD36) ,and Niemann-Pick type C1-like 1 protein ( NPC1L1) proteins in small intestinal tissues were detected by Western blot.
Results :
The results of HE and Oil red O staining of liver tissues showed that liver fat deposition in LSS gene heterozygous knockout mice was lower than that in wild-type mice in the high-fat diet group.The total cholesterol content in intestinal tis- sue of LSS gene heterozygous knockout mice decreased ( P <0. 01) ,but no morphological differences were ob- served between the two groups of mice by HE staining of intestinal tissues.The gastrointestinal motility function of LSS gene heterozygous knockout mice did not show significant changes.The intestinal permeability of LSS gene het- erozygous knockout mice in the high-fat diet group decreased as detected by Evans blue ( P<0. 05) .The expres- sion levels of Claudin-5 protein in the intestinal tissue of LSS gene heterozygous knockout mice in the high-fat diet group increased ( P <0. 05 ) ,while the expression of LSS protein in the intestinal tissues of LSS heterozygous knockout mice decreased ( P <0. 05) .
Conclusion
In the NAFLD model induced by a high-fat diet,LSS gene heterozygous knockout reduces liver fat deposition induced by a high-fat diet and improves intestinal barrier function by regulating cholesterol metabolism in intestinal tissues and up-regulating the expression of Claudin-5.
4.Changes in behavior and spatial memory of C57BL/6J mice of different ages
Zhen Yang ; Hongmei Bai ; Weikang Hu ; Mingcong Li ; Xiaoli Jiang ; Chaoyang Zhang ; Zihan Wang ; Wenjing Zhou ; Qingya He ; Jian Zhong ; Shengquan Zhang
Acta Universitatis Medicinalis Anhui 2025;60(8):1410-1417
Objective :
To explore the changes in behavior and spatial memory of C57BL/6J female mice of different ages (youth , middle-aged , and elderly) .
Methods:
C57BL/6J female mice were divided into female youth group (YG group) , female middle-aged group ( MG group) and female elderly group ( OG group) according to age. The Morris water maze test measured spatial memory ability , and the open field and elevated cross maze test observed activity level and anxiety level. Western blot was used to determine the protein expressions of CREB , CaMKⅡ(pan) and CaMKⅡ(p) in the hippocampus of the brain tissues of female mice in each group.
Results:
Compared with the YG group , the weight of the MG group and the OG group significantly increased (P < 0. 01 , P < 0. 001) . Compared with the OG group , the third quadrant escape latency and the number of crossings in the YG group and MG group were shortened , and the difference was not statistically significant. Compared with the OG group , there was a statistically significant difference in the exercise speed in the open field of the YG group (P < 0. 01) , there was no significant difference in the movement speed in the open field of the MG group , the number of entries into the central zone significantly increased in the MG group ( P < 0. 05 ) , and there was no significant difference in the number of entries in the YG group (P > 0. 05) . Compared with the OG group , the YG group had a statistically significant difference in the elevated cross maze (P < 0. 05) , the MG group had no statistically signif- icant difference in the elevated cross maze , and the number of closed arm entries in the YG group and MG group significantly increased (P < 0. 001 , P < 0. 01) . Compared with the YG group , the relative expression level of CaMKⅡ(pan) in the OG group was statistically significant ( P < 0. 05 ) , while the relative expression level of CaMKⅡ(pan) in the MG group was not statistically significant ( P > 0. 05) .
Conclusion
With the increase of age , the weight of C57BL/6J female mice gradually increased , the activity level and desire to explore gradually de- creased , the spatial memory ability also declined , and the anxiety level and anxiety-like behavior increased. This study helps to reveal the effect of age on the activity level and cognitive function of females , and provides a refer- ence for studying cognitive and memory decline in older females.
5.Lysine Succinylation:A New Perspective in the Treatment of Cardiovascular Diseases
Yang MENG ; Yuyang MA ; Mingcong CHEN ; Yang ZHANG ; Mingyi ZHAO
Acta Academiae Medicinae Sinicae 2024;46(3):435-443
With the continuous development of identification technologies such as mass spectrometry,omics,and antibody technology,post-translational modification(PTM)has demonstrated increasing potential in medical research.PTM as a novel chemical modification method provides new perspectives for the research on dis-eases.Succinylation as a novel modification has aroused the interest of more and more researchers.The available studies about succinylation mainly focus on a desuccinylase named sirtuin 5.This enzyme plays a key role in mod-ification and has been preliminarily explored in cardiovascular studies.This paper summarizes the influencing fac-tors and regulatory roles of succinylation and the links between succinylation and other PTMs and reviews the re-search progress of PTMs in the cardiovascular field,aiming to deepen the understanding about the role of this modification and give new insights to the research in this field.
6.The study of molecular mechanism of regulation of IL-10 on proliferation and differentiation of HaCaT cells
Xueli Yin ; Bo Jia ; Li Liu ; Mingcong Li ; Jun Zhang ; Zhen Yang ; Hongmei Bai ; Weikang Hu ; Sumei Zhang ; Shengquan Zhang
Acta Universitatis Medicinalis Anhui 2023;58(6):890-895
Objective:
To investigate the effects of interleukin ( IL) -10 on the proliferation of HaCaT cells and CaCl2 induced expression of differentiation markers and its possible molecular mechanisms.
Methods:
HaCaT cells were treated with various concentrations of IL-10 (0,3,10,30 ng / ml) for different time (0,24,48,72 h) ,cell proliferation was measured using MTS,and cell cycle was determined by flow cytometry.HaCaT cells were pretreated with IL-10 (final concentration 10 ng / ml) for 1 h,then incubated with or without CaCl2 (final concentration 1. 2 mmol / L) for 24,48,72 h ,Western blot was performed to detect the effect of IL-10 on the expression of HaCaT keratinocyte differentiation markers.After pretreatment of HaCaT cells with PD98059,an inhibitor of mitogen-activated kinase-ERK1 /2,and LY294002,an inhibitor of phosphatidylinositol kinase-serine / threonine kinase (PI3K-AKT) ,the total RNA and proteins were extracted separately,real time quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to examine the influence of IL-10 on the expression of differentiation markers (Keratin1,Keratin5,Involucrin) .
Results :
MTS results revealed that IL-10 (30 ng / ml and lower doses) did not alter the proliferation of HaCaT cells in 72 h.Flow cytometry analysis demonstrated that IL-10 had no significant influence on cell cycle progression.The results of Western blot showed that IL-10 upregulated the expression of differentiation markers Involucrin,while there was no significant effect on Keratin1 and Keratin5 .Mechanism research analysis demonstrated that IL-10 could activate ERK1 /2 and AKT ,increase their phosphorylation levels ; RT-qPCR and Western blot results showed that PD98059 and LY294002 partially blocked IL-10 induced Involucrin expression.
Conclusion
At a particular concentration range,IL-10 has little effect on HaCaT proliferation ,but it partially upregulates the expression of differentiation marker Involucrin via the MAPKs-ERK1 /2 and PI3K-AKT pathways.
7.Influential factors of acute kidney injury after acute myocardial infarction and its effect on adverse cardiovascular events during hospitalization
Mingcong YAN ; Chunlin YIN ; Yanli ZHANG
Chinese Journal of Postgraduates of Medicine 2022;45(1):18-24
Objective:To observe the incidence and risk factors of acute kidney injury (AKI) in patients with acute myocardial infarction (AMI), and explore the effect of AKI on the adverse cardiovascular events during hospitalization.Methods:The clinical data of 1 286 first-episode patients with AMI from December 2014 to December 2017 in Xuanwu Hospital, Capital Medical University were retrospectively analyzed. The basic information, laboratory test results and used drug were collected, the incidences of AKI and adverse cardiovascular events during hospitalization were recorded.Results:Among 1 286 patients with AMI, 113 cases had AKI, the incidence of AKI was 8.79%. There were statistical differences in gender composition, age, Killip classification>1 grade, hypertension, diabetes, smoking history, left ventricular ejection fraction (LVEF)<56%, baseline estimated glomerular filtration rate (eGFR)<71.5 ml/(min·1.73 m 2), global registry of acute coronary events score (GRACE score) ≥176 scores, hemoglobin<128 g/L, interleukin-6 (IL-6) ≥35.74 ng/L, brain natriuretic peptide (BNP) ≥981 ng/L, thrombolysis in myocardial infarction score (TIMI score) ≥5 scores, high-sensitivity C-reactive protein (hs-CRP) ≥8.44 mg/L, serum creatinine at admission ≥90 μmol/L, heart rate ≥75 times/min and contrast agent dosage/eGFR ≥1.92 between patients with AKI and patients without AKI ( P<0.01 or<0.05). Multivariate Logistic regression analysis result showed that diabetes, LVEF<56%, baseline eGFR< 71.5 ml/(min·1.73 m 2) and hs-CRP ≥8.44 mg/L were the independent risk factors of AKI in patients with AMI ( OR = 2.99, 0.38, 0.30 and 2.48; 95% CI 1.31 to 6.84, 0.16 to 0.88, 0.12 to 0.78 and 1.07 to 5.75; P = 0.010, 0.024, 0.013 and 0.035). The hospital mortality, incidence of adverse cardiovascular events during hospitalization and length of hospital stay in patients with AKI were significantly higher than those in patients without AKI: 11.50% (13/113) vs. 2.39% (28/1 173), 22.12% (25/113) vs. 8.40% (99/1 173) and (12±8) d vs. (10±6) d, and there were statistical differences ( P<0.01). Multivariate Logistic regression analysis result showed that AKI was the independent risk factors of death and adverse cardiovascular events during hospitalization in patients with AMI ( OR = 5.32 and 3.08, 95% CI 2.67 to 10.59 and 1.89 to 5.03, P<0.01). Conclusions:The incidence of AKI is high in patients with AMI, and previous diabetes history, LVEF<56%, eGFR<71.5 ml/(min·1.73 m 2), hs-CRP≥8.44 mg/L are the independent risk factors of AKI in patients with AMI. The occurrence of AKI after AMI can increase incidence of adverse cardiovascular events during hospitalization and hospital mortality, and prolong the hospital stay.
8.Screening of differential genes and pathways in C57BL/6J mice of different genders based on transcriptome sequencing
Yuhao Li ; Yanling Li ; Xueli Yin ; Xiaomei Sun ; Jun Zhang ; Mingcong Li ; Li Liu ; Sumei Zhang ; Shengquan Zhang
Acta Universitatis Medicinalis Anhui 2022;57(5):742-747
Objective:
To screen the differential genes and pathways of female and male C57 BL/6 J mice by transcriptome sequencing, and to lay a foundation for further exploring the molecular mechanism of behavioral differences between male and female mice.
Methods:
8-week-old female and male mice of the C57 BL/6 J strain were completely isolated from the mouse hippocampus, and total RNA was extracted and reverse transcribed to construct a cDNA library, and 50 single-ended mice were generated on the BGIseq500 platform(BGl-Shenzhen, China) The base reads were transcriptome sequenced. After sequencing, based on GO and KEGG databases, combined with the phyper function in R language to screen, correct and enrich the data, calculatePvalue, and then perform FDR correction onPvalue to obtainQvalue and analyze different annotated genes based on the hypergeometric test method. The expression status was analyzed, and the differential genes and pathways in the hippocampus of mice of different genders were screened out.
Results:
Through the comparison of male and female differences, 325 differential genes were screened, including 233 up-regulated genes and 92 down-regulated genes. The functions of these differential genes were mainly enriched in long-term potentiation(LTP), calcium signal pathway, nicotine addiction and other processes. There were 362 junctions and 1 703 interaction edges in the female-male differential gene interaction network, and 10 core genes selected: lysine demethylase 5 D(Kdm5 d), cyclin-dependent kinase 5(Cdkl5), Cell output mediator(Cle1), dopamine receptor 6 a3(Slc6 a3), cassette transcription factor(Fox), precursor mRNA processing factor 4 B(Prpf4 b), glycine receptor 4(Glur4), calcium ion receptor 2(Camk2), DNA and RNA binding protein family(Son), serotonin receptor 5 b(Htr5 b).
Conclusion
The selected differential genes and signal pathways may lay the foundation for explaining the molecular mechanism of the differences in behavior between male and female mice.
9.Thinking on Integrating Health Ethics into Health Education and Prospect of Its Practice Path
Jingwen LI ; Mingcong TANG ; Xi ZHANG ; Yu CHEN ; Siyang YE ; Shuangmiao WANG
Chinese Medical Ethics 2022;35(12):1399-1403
Since the reform and opening-up, China’s health care has made great progress, and the level of national health literacy has steadily improved. However, there is still a disconnect between the health literacy and healthy behavior of Chinese residents, and traditional health education has little effect on behavior change. Based on the limitations of current traditional health education on improving health level of the whole people, this paper explored more effective education methods, deeply discussed how to integrate health ethics into health education to achieve the purpose of effectively promoting individual health behaviors. At the same time, this paper systematically expounded how the theory of behavioral economics provides theoretical support for the rationality and feasibility of health ethics education to promote healthy behavior, further explained the internal psychological mechanism of health ethics education affecting people’s healthy behavior, and provided feasible solutions for how to integrate health ethics into the new model of health education in practical application. To sum up, the integration of health ethics into health education is conducive to disseminating health concepts, improving health literacy, as well as promoting health behaviors, and then promoting the effective implementation of individual health and Healthy China.
10.Effects of lanosterol synthase inhibitor on differentiation and apoptosis of KCs
Li Liu ; Yanan Gu ; Mingcong Li ; Yixuan Huang ; Shengquan Zhang
Acta Universitatis Medicinalis Anhui 2022;57(2):173-177
Objective :
To investigate the effects of RO or incubation with cholesterol ( CH) on the differentiation and apoptosis of KCs.
Methods :
Ca2 + ( 1. 8 mmol/L) was added to KCs for 1 day after co - culture of RO alone or combined with CH in KCs for different time. The expression of Involucrin (INV) and Loricrin in KCs was analyzed by Western blot. After co⁃culture of RO alone or combination with CH in KCs for different time , the apoptotic changes of KCs cells were detected by flow cytometry and the expression of apoptotic proteins Bax and Bcl⁃2 were verified by Western blot.
Results :
RO down⁃regulated the expression of Ca2 + induced differentiation marker protein INV , but weakly inhibited the expression of Loricrin , while CH showed no antagonistic effect on RO. RO induced
apoptosis of KCs cells in a time dependent manner. CH could antagonize the apoptotic effect of RO on KCs ; the expression of Bcl⁃2 and Bax was inhibited when RO was applied alone , CH could partially antagonize the inhibition effect of RO on Bcl⁃2 expression , but had no significant effect on Bax; however, RO reduced the ratio of Bcl⁃2/Bax in a time⁃dependent manner, and CH partially weakened the effect of RO on the ratio of Bcl⁃2/Bax.
Conclusion
RO may inhibit KCs differentiation and induce KCs cell apoptosis by down⁃regulating the expression of Loricrin and Bcl⁃2.


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