1.Effect and Mechanism of Total Saponins from Panax Japonicus on White Adipose Tissue Browning/Brown Adipose Tissue Activation in High-fat Diet-induced Mice
Shuwen WANG ; Yaqi HU ; Rui WANG ; Yifan ZHANG ; Mengzhen XUE ; Yaqi WANG ; Fangqi XIA ; Leiqi ZHU ; Chengfu YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):71-78
ObjectiveTo investigate the effect and mechanism of total saponins from Panax japonicus (TSPJ) on white adipose tissue (WAT) browning/brown adipose tissue (BAT) activation in C57BL6/J male mice fed on a high-fat diet (HFD). MethodThirty-two C57BL6/J male mice (8-week-old) were randomly divided into a normal group, a model group, a low-dose TSPJ group, and a high-dose TSPJ group. The mice in the low-dose and high-dose TSPJ groups were given TSPJ for four months by gavage at 25, 75 mg·kg-1·d-1, respectively, and those in the other groups were given 0.5% sodium carboxymethyl cellulose (CMC-Na) accordingly. After four months of feeding, all mice were placed at 4 ℃ for acute cold exposure, and the core body temperature was monitored. Subsequently, all mice were sacrificed, and BAT and inguinal WAT (iWAT) were separated rapidly to detect the corresponding indexes. Hematoxylin-eosin (HE) staining was used to observe the morphological changes in each group. The effect of TSPJ on the mRNA expression of uncoupling protein 1 (UCP1), fatty acid-binding protein 4 (FABP4), cytochrome C (CytC), PR domain-containing protein 16 (PRDM16), elongation of very long chain fatty acids protein 3 (ELOVL3), peroxisome proliferator-activated receptor γ (PPARγ), and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) in iWAT and BAT was detected by Real-time polymerase chain reaction (Real-time PCR). Western blot was also used to detect the protein expression of UCP1, PRDM16, PPARγ, and PGC-1α in BAT and iWAT of each group. The effect of TSPJ on UCP1 expression in BAT and iWAT was detected by immunohistochemistry. Result① Compared with the model group, TSPJ could decrease the body weight and proportions of iWAT and BAT in the HFD-induced mice (P<0.05, P<0.01). ② The body temperature of mice in the model group decreased compared with that in the normal group in the acute cold exposure tolerance test (P<0.05). The body temperature in the high-dose TSPJ group increased compared with that in the model group (P<0.01). ③ Compared with the normal group, the model group showed increased adipocyte diameter in iWAT and BAT and decreased number of adipocytes per unit area. Compared with the model group, the TSPJ groups showed significantly reduced cell diameter and increased number of cells per unit area, especially in the high-dose TSPJ group. ④ Compared with the normal group, the model group showed decreased mRNA expression of FABP4, UCP1, CytC, PRDM16, ELOVL3, PGC-1α, and PPARγ in adipose tissues of mice (P<0.05, P<0.01). Compared with the model group, after intervention with TSPJ, the mRNA expression was significantly up-regulated (P<0.05, P<0.01). ⑤ Compared with the normal group, the model group showed decreased protein expression of UCP1, PRDM16, PPARγ, and PGC-1α in adipose tissues of mice (P<0.05, P<0.01). Compared with the model group, after intervention with TSPJ, the protein expression increased significantly (P<0.05, P<0.01). ConclusionTSPJ could induce the browning of iWAT/BAT activation and enhance adaptive thermogenesis in obese mice induced by HFD. The underlying mechanism may be attributed to the activation of the PPARγ/PGC-1α signaling pathway.
2.Protective Effect and Mechanism of Proanthocyanidin B2 Against H2O2-induced Oxidative Damage and Apoptosis of Astrocytes
Shuwen YUAN ; Yiwei DONG ; Jian LIU ; Yajie LIANG ; Jianjun HUANG ; Baoguo XIAO ; Qing WANG ; Cungen MA
Chinese Journal of Modern Applied Pharmacy 2024;41(6):727-735
OBJECTIVE
To investigate the protective effect proanthocyanidin B2(PC-B2) on oxidative damage and apoptosis of mouse astrocytes(AS) induced by hydrogen peroxide(H2O2) and its mechanism.
METHODS
AS were isolated and cultured from neonatal C57BL/6 mice(1−3 d). The optimal concentration of H2O2 and PC-B2 was divided into four groups: normal group, normal+PC-B2 group(100 μg·mL‒1 PC-B2 treated for 24 h), H2O2 model group(200 μmol·L‒1 H2O2 treated for 24 h), PC-B2 group(200 μmol·L‒1 H2O2 and 100 μg·mL‒1 PC-B2 treated for 24 h). The cell viability of each group was detected by CCK-8 method. Cytotoxicity was detected by LDH method. The antioxidant capacity was detected by ABTS and DPPH. The content of MDA and the activity of SOD, CAT and GSH-Px were detected by ELISA kit. Detection of apoptosis in each group was done by TUNEL staining. The mRNA and protein expression levels of Bax, Bcl-2, Caspase-3, Akt/Stat3, p-Akt, p-Stat3 and Nrf2/HO-1 in AS were detected by RT-PCR and Western blotting, respectively.
RESULTS
PC-B2 could significantly enhance cell viability and inhibit AS apoptosis. Compared with the H2O2 model group, PC-B2 intervention could significantly reduce the content of LDH and MDA in AS, and increase the activity of SOD, CAT and GSH-Px. PC-B2 intervention could inhibit the mRNA and protein expression of Bax and Caspase-3, and up-regulate the mRNA and protein expression of Akt/Stat3, Bcl-2, Nrf2/HO-1.
CONCLUSION
PC-B2 can enhance the antioxidant capacity of AS through Akt/Stat3 and Nrf2/HO-1 pathways, therefore reduce H2O2-induced AS oxidative damage and apoptosis.
3.Long non-coding RNA AW112010 improves insulin resistance in adipocytes of aging mice through the miR-204/POU2F2 axis
Rui WANG ; Shuwen WANG ; Yifan ZHANG ; Yaqi HU ; Qi YUAN ; Yuan WEN ; Xiaoling CHEN ; Ting LU ; Ying ZHENG ; Zhiyong LIN ; Mengzhen XUE ; Yaqi WANG ; Fangqi XIA ; Leiqi ZHU ; Chengfu YUAN
Chinese Journal of Endocrinology and Metabolism 2024;40(1):44-52
Objective:To investigate whether long non-coding RNA(lncRNA) AW112010 can improve insulin resistance in aging adipocytes through the miR-204/POU2F2 signaling pathway.Methods:In vivo experiment: C57BL/6 mice were divided into young control group(4 months old) and aging model group(18 months old) based on body weight. The expression levels of AW112010, miR-204-5p, POU2F2, aging related indicators(p16, p21), and insulin signaling pathway genes [insulin receptor(INSR), insulin receptor substrate 1(IRS1), phosphatidylinositol kinase(PI3K), protein kinase B(AKT)] in epididymal adipose tissue were detected using real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting. In vitro experiment: Using adriamycin(ADR) to induce 3T3-L1 aging adipocyte model, β-gal staining was used to observe cellular senescence, and miR-204 inhibitor and miR-204 mimic small interfering RNA were successfully constructed and transfected into 3T3-L1 adipocytes. Results:RT-qPCR and Western blot results showed that compared with the young group, the expression of AW112010 in the adipose tissue of aging mice was increased, while the expression of miR-204-5p was decreased. The expressions of POU2F2, p16, and p21 in the adipose tissue of aging mice were increased, while the expressions of INSR, IRS1, PI3K, GLUT4 mRNA and protein were decreased. The β-gal stainging results showed that the number of 3T3-L1 senescent adipocytes induced by ADR was significantly increased, and the expression levels of AW112010, POU2F2, p16, and p21 in ADR-induced senescent adipocytes were increased compared with the control group, while the expression levels of miR-204-5p, INSR, IRS1, PI3K, GLUT4 were decreased, and remaining glucose in the culture medium was increased. Compared with control, overexpression of miR-204 resulted in decreased expressions of aging indicators p16, p21, and target gene POU2F2 while the expressions of INSR and GLUT4 were increased.Conclusion:Upregulation of lncRNA AW112010 in adipocytes of aging mice may induce insulin resistance by targeting miR-204-5p/POU2F2/IRS1.
4.Effect of dynamic changes of microglia on oligodendrocytes in animal model of multiple sclerosis
Shuwen YUAN ; Kerong QI ; Qingqing LIU ; Xue CHEN ; Jianchao SI ; Shengchang YANG
Chinese Journal of Neurology 2024;57(6):669-677
Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous syestem (CNS) triggered by an autoimmune mechanism, which is the main cause of neurological disability in young people. In MS, a variety of glial cells participate in the pathogenesis and development of the disease, and oligodendrocytes (OL) and myelin cytes are destroyed by autoimmune mediated inflammation. Impaired OL production of oligodendrocyte progenitor cells (OPC) leads to persistent demyelination, myelin fragment accumulation and axonal injury, with clinical manifestation of CNS disability. Microglia (MG), which is involved in the inflammatory response and removal of myelin debris, plays a pivotal role in OPC differentiation and OL maturation, thereby directly or indirectly influencing the process of myelin regeneration. An interactive mechanism exists between these 2 components. The comprehension of the interrelationship between the 2 factors can provide enhanced insights into the pathogenesis of MS and facilitate the development of novel therapeutic strategies, thereby addressing pivotal scientific challenges encountered in clinical management of MS. Therefore, this review summarizes the dynamic changes of microglia in MS and its classic animal models, explores the internal relationship between MG and OL in this process, and focuses on the beneficial effects of targeted regulation of microglia, thereby promoting OPC differentiation and myelin regeneration, in order to provide a new research direction for MS treatment.
5.Research progress in changes of intestinal flora after spinal cord injury and their effects on spinal neuroinflammation
Haixia CHEN ; Hongru LI ; Jingyi LIU ; Zhifang XU ; Shuwen LIU ; Yuan YANG ; Yang CHEN ; Yu LUO ; Yinjie CUI
Journal of Jilin University(Medicine Edition) 2024;50(6):1751-1756
The gut microbiota is a vast microbial ecosystem,specifically present in the organism and plays an important regulatory role in the body's health or disease state together with its metabolites.After spinal cord injury,the complex pathophysiology at the site of trauma makes axonal regeneration difficult,and the autonomic motor dysfunction induced by spinal cord injury disrupts gastrointestinal function and causes gut microbiota imbalance.The previous clinical outcomes of neurorepair strategies after spinal cord injury have not been ideal.The dysregulated gut microbiota and neuroinflammation after spinal cord injury are closely associated with the prognosis of the patients.The potential mechanisms by which the gut microbiota may influence the neuroinflammation after spinal cord injury may include the activation of gut-associated lymphoid tissue and disruption of the intestinal barrier by the imbalanced microbiota,and gut microbiota and its metabolites such as lipopolysaccharides(LPS),short chain fatty acids(SCFAs),5-hydroxytryptamine(5-HT),and tryptophan,as well as immune cells,inflammatory factors,and neurotransmitters the local inflammatory response in the spinal cord through the circulatory system.This paper revews the studies on the changes in gut microbiota after spinal cord injury and their effects on the spinal cord neuroinflammation,providing new targets and new ideas for improving the neuroinflammation after spinal cord injury.
6.Application of wearable visual training system based on extended reality glasses in patients after macular hole surgery
Jing YUAN ; Xingchang WANG ; Xiquan SUN ; Huiguang JIAO ; Qian WANG ; Jianxiong YU ; Biyue TU ; Xixi YAN ; Zhen ZHAO ; Yanxia TONG ; Shuwen ZHANG
Chinese Journal of Experimental Ophthalmology 2024;42(12):1142-1147
Objective:To evaluate the short-term rehabilitation effect of wearable visual training devices based on extended reality (XR) glasses in patients after macular hole surgery.Methods:A self-controlled study was conducted.Eleven patients with monocular low vision after macular hole surgery were recruited at Renmin Hospital of Wuhan University from October 2022 to March 2024.All patients underwent biofeedback training for 3 months using the independently developed visual rehabilitation training glasses LOOKBON T10.The LogMAR best corrected visual acuity (BCVA), retinal sensitivity, effective fixation rate, fixation stability, reading speed, vertical metamorphopsia (MV), horizontal metamorphopsia (MH), and Chinese version of the visual-related quality of life assessment form (CVRQoL-25) were compared before and after training.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University (No.WDRY2024-K263).Written informed consent was obtained from each subject.Results:After training, the patients' BCVA, retinal sensitivity, effective fixation rate, fixation stability, and reading speed were 0.69±0.19, (21.61±2.75)db, (92.43±4.06)%, (93.09±4.31)%, and (104.82±21.85) characters/minute, respectively, which were significantly improved compared to 0.85±0.28, (17.71±3.17)db, (31.83±19.05)%, (32.35±19.12)%, and (69.64±20.17) characters/minute before training ( t=5.253, -5.987, -11.561, -12.003, -11.682; all at P<0.001).After training, MV and MH were (0.29±0.20)° and (0.21±0.24)°, respectively, which were significantly reduced compared to pre-training (0.44±0.24)° and (0.43±0.41)° ( t=9.238, 4.068; both at P<0.01).After training, the CVRQoL-25 score was 1 193.18±229.43, which was significantly higher than pre-training 947.73±203.86 ( t=-11.687, P<0.001). Conclusions:The application of wearable visual training equipment based on XR glasses can effectively improve the visual function of patients with poor visual function recovery after macular hole surgery, and enhance their quality of life.
7.Factors influencing pulmonary infection in elderly neurocritical patients and their predictive values: a data analysis for consecutive four-year
Jia YUAN ; Ying LIU ; Di LIU ; Difen WANG ; Feng SHEN ; Xu LIU ; Shuwen LI ; Dehua HE
Chinese Critical Care Medicine 2023;35(1):66-70
Objective:To analyze the factors influencing pulmonary infections in elderly neurocritical patients in the intensive care unit (ICU) and to explore the predictive value of risk factors for pulmonary infections.Methods:The clinical data of 713 elderly neurocritical patients [age ≥ 65 years, Glasgow coma score (GCS) ≤ 12 points] admitted to the department of critical care medicine of the Affiliated Hospital of Guizhou Medical University from 1 January 2016 to 31 December 2019 were retrospectively analyzed. According to whether or not they had HAP, the elderly neurocritical patients were divided into hospital-acquired pneumonia (HAP) group and non-HAP group. The differences in baseline data, medication and treatment, and outcome indicators between the two groups were compared. Logistic regression analysis was used to analyze the factors influencing the occurrence of pulmonary infection.The receiver operator characteristic curve (ROC curve) was plotted for risk factors and a predictive model was constructed to evaluate the predictive value for pulmonary infection.Results:A total of 341 patients were enrolled in the analysis, including 164 non-HAP patients and 177 HAP patients. The incidence of HAP was 51.91%. According to univariate analysis, compared with the non-HAP group, mechanical ventilation time, the length of ICU stay and total hospitalization in the HAP group were significantly longer [mechanical ventilation time (hours): 171.00 (95.00, 273.00) vs. 60.17 (24.50, 120.75), the length of ICU stay (hours): 263.50 (160.00, 409.00) vs. 114.00 (77.05, 187.50), total hospitalization (days): 29.00 (13.50, 39.50) vs. 27.00 (11.00, 29.50), all P < 0.01], the proportion of open airway, diabetes, proton pump inhibitor (PPI), sedative, blood transfusion, glucocorticoids, and GCS ≤ 8 points were significantly increased than those in HAP group [open airway: 95.5% vs. 71.3%, diabetes: 42.9% vs. 21.3%, PPI: 76.3% vs. 63.4%, sedative: 93.8% vs. 78.7%, blood transfusion: 57.1% vs. 29.9%, glucocorticoids: 19.2% vs. 4.3%, GCS ≤ 8 points: 83.6% vs. 57.9%, all P < 0.05], prealbumin (PA) and lymphocyte count (LYM) decreased significantly [PA (g/L): 125.28±47.46 vs. 158.57±54.12, LYM (×10 9/L): 0.79 (0.52, 1.23) vs. 1.05 (0.66, 1.57), both P < 0.01]. Logistic regression analysis showed that open airway, diabetes, blood transfusion, glucocorticoids and GCS ≤ 8 points were independent risk factors for pulmonary infection in elderly neurocritical patients [open airway: odds ratio ( OR) = 6.522, 95% confidence interval (95% CI) was 2.369-17.961; diabetes: OR = 3.917, 95% CI was 2.099-7.309; blood transfusion: OR = 2.730, 95% CI was 1.526-4.883; glucocorticoids: OR = 6.609, 95% CI was 2.273-19.215; GCS ≤ 8 points: OR = 4.191, 95% CI was 2.198-7.991, all P < 0.01], and LYM, PA were the protective factors for pulmonary infection in elderly neurocritical patients (LYM: OR = 0.508, 95% CI was 0.345-0.748; PA: OR = 0.988, 95% CI was 0.982-0.994, both P < 0.01). ROC curve analysis showed that the area under the ROC curve (AUC) for predicting HAP using the above risk factors was 0.812 (95% CI was 0.767-0.857, P < 0.001), with a sensitivity of 72.3% and a specificity of 78.7%. Conclusions:Open airway, diabetes, glucocorticoids, blood transfusion, GCS ≤ 8 points are independent risk factors for pulmonary infection in elderly neurocritical patients. The prediction model constructed by the above mentioned risk factors has certain predictive value for the occurrence of pulmonary infection in elderly neurocritical patients.
8.Changes of Intestinal Microbiota in Type 2 Diabetic Diarrhea Patients with Different Syndromes Based on High-throughput Sequencing
Yujin WANG ; Shuwen DOU ; Yuan ZHOU ; Qingying WANG ; Conge TAN ; Xiangdong WANG ; Wenwen XING ; Ying YAN ; Yanjin SU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(2):125-132
ObjectiveTo investigate the changes of intestinal microbiota in type 2 diabetic diarrhea (T2DD) patients with dampness-heat syndrome and spleen-kidney deficiency syndrome. MethodT2DD patients who were admitted to the Department of Endocrinology Ⅰ of the Affiliated Hospital of Shaanxi University of Chinese Medicine from March 2020 to May 2021 were selected, including 14 patients with type 2 diabetes mellitus (T2DM), 12 T2DD patients with dampness-heat syndrome, and 13 T2DD patients with spleen-kidney deficiency syndrome. Twelve healthy subjects receiving medical examination were selected as control group. Their body mass index (BMI), fasting plasma glucose (FPG), 2 h postprandial blood glucose (2 h PBG) and glycosylated hemoglobin (HbA1c) were compared. Fecal samples were collected for DNA extraction to build a database. High-throughput 16S rDNA sequencing was used to compare the composition of intestinal microbiota and the differential bacteria among the four groups. ResultCompared with the conditions in control group, the levels of FPG, 2 h PBG and HbA1c in the other groups were increased (P<0.05). Alpha diversity showed no significant difference in species richness, evenness and diversity of intestinal microbiota among the groups. Beta diversity indicated that intestinal microbiota tended to be consistent in each group, and there was no marked difference between groups. The top 5 phylum by relative abundance were Bacteroidetes,Proteobacteria, Firmicutes, Actinobacteria, and Fusobacteria, among which,Bacteroidetes, Proteobacteria and Firmicutes were dominant. Compared with the control group, the three diabetic groups had elevated relative abundance of Bacteroidetes while decreased relative abundance of Firmicutes. The relative abundance of Actinomycetes in spleen-kidney deficiency T2DD group was significantly higher than that in the other groups, and the relative abundance of Firmicutes and Fusobacteria in the dampness-heat T2DD group was significantly lower than that in the other groups. At the genus level, the top 10 bacteria by relative abundance were Phocaeicola, Bacteroides, Pseudescherichia, Prevotella, Bifidobacterium, Faecalibacterium, Fusobacterium, Roseburia, Citrobacter, and Cetobacterium. LEfSe analysis revealed that the relative abundance of Prevotella, Mediterraneibacter, Parabacteroides, and Fusicatenibacter in diabetic patients was remarkably higher than that in healthy patients. Bacteroides and Sutterella might be the characteristic microbiota of T2DD patients with dampness-heat syndrome, while Faecalibacterium, Limosilactobacillus, Eubacterium, Gemmiger, Enterocloster, Alistipes, Parasutterella and Oscillibacter might be the characteristic microbiota of T2DD patients with spleen-kidney deficiency syndrome. ConclusionBacteroides and Parasutterella might be the characteristic microbiota of T2DD patients with dampness-heat syndrome and spleen-kidney deficiency syndrome, respectively. This paper provided reference for studying the mechanism, diagnosis and treatment of modern traditional Chinese medicine for T2DD of dampness-heat type and spleen-kidney deficiency type.
9.Association between platelet/lymphocyte ratio and frequent peritoneal dialysis-associated peritonitis in peritoneal dialysis patients
Jing YUAN ; Yuqi YANG ; Lu LIU ; Fangfang YU ; Shuwen QIE ; Li YANG ; Yan ZHA
Chinese Journal of Nephrology 2021;37(4):327-332
Objective:To explore the association between platelet/lymphocyte ratio (PLR) and frequent peritoneal dialysis (PD) - associated peritonitis (PDAP) in PD patients.Methods:The data of PD patients with PDAP from Guizhou Provincial People's Hospital between January 2015 and June 2019 were analyzed retrospectively. The patients were divided into mono group (only once PDAP occurred in one year) and frequent group (2 or more PDAP occurred in one year) according to the frequency of PDAP. The demographic data including gender, age, height and weight, the clinical data including blood pressure, duration of PD, causes of peritonitis, the laboratory data at the first time of PDAP and the prognosis of PDAP were compared between two groups. Logistic regression analysis method was applied to analyze the relationship between PLR and frequent PDAP. The predictive power of PLR was evaluated by receiver operating characteristic curve (ROC).Results:A total of 78 PD patients with PDAP were enrolled, including 53 males and 25 females, with average age of 45.2 years. The total person-year was 765.1 person-years and the incidence of peritonitis was 0.10 case/person-year during the median follow-up of 16 months. All patients were divided into two groups: 53 patients in mono group and 25 patients in frequent group. Compared with mono group, the patients in frequent group had lower body mass index, longer dialysis duration, higher systolic blood pressure level, higher PLR level, lower uric acid level, and higher rate of drug-resistant bacteria in peritoneal effusion (all P<0.05). The extubation rate of the frequent group was 44.0%(11/25), which was significantly higher than that [15.1%(8/53)] of mono group ( P<0.05). Multivariate logistic regression analysis showed that higher PLR level was an independent related factor for frequent PDAP( OR=1.006, 95% CI 1.002-1.010, P=0.003), and the area under the ROC curve of PLR was 0.783(95% CI 0.663-0.904, P<0.001). Conclusions:High PLR level is an independent related factor of frequent PDAP for PD patients, and PLR can be a potential predictor of frequent PDAP.
10.Genetic analyzing for a Chinese intellectual disability pedigree with ARX gene mutation
Lina CUI ; Shuwen XIN ; Aolong YUAN ; Jingshang LYU ; Rongxiang LI ; Shaozhe YANG ; Jing XU ; Xuna XING ; Yaya YANG ; Jiajia YANG ; Xiuhong FU
Chinese Journal of Laboratory Medicine 2021;44(3):228-232
Objective:To investigate the molecular mechanism of the disease based on the clinical characterization and genetic mutation analysis in a family with intellectual disability.Methods:The proband with intellectual disability was diagnosed at Luohe Central Hospital in December 2019. Peripheral blood samples were collected from four family members. Whole exome sequencing (WES) was used to screen the pathological mutations. Then the PCR and Sanger sequencing were used to verify the selected mutations and combine the relevant database to analyze variation loci.Results:We infer that the ARX c.1162 A>G was co-segregated with the phenotype of the family based on the results of WES. The results of sanger sequencing and WES are consistent. The mother of the proband is the carrier of the mutation. There is no mutation frequency reported in the healthy population. The mutation of the ARX c.1162A>G is harmful inferred by a variety of bioinformatics software. Combined with the phenotypic analysis of OMIM database, we infer the phenotype caused by the mutation is consistent with the patients in the family.Conclusion:The mutation of the ARX c.1162 A>G may be the cause of the intellectual disability in the family affected. And the variant has not been reported in China.


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