1.Study of Improvement Effects and Mechanism of Saposhnikovia Divaricata Extract on IgE Sensitized RBL-2H3 Cells
Sisi CHEN ; Limei QIAN ; Yanchun CHEN
Journal of Zhejiang Chinese Medical University 2024;48(2):138-146
[Objective]To explore the degranulation effect and mechanism of Saposhnikovia divaricata(SD)extract on rat basophilic leukemia cell line RBL-2H3 cell.[Methods]Methylthialazole tetrazolium(MTT)test was used to select the concentrations in the subsequent experiments based on impact of 5,25,50,100,200,400 μg·mL-1 SD extract on the activity of RBL-2H3 cells.Immunoglobulin E(IgE)induction was used to establish RBL-2H3 cell degranulation model.Blank control group,model group,low dose SD extract group(5 μg·mL-1),medium dose SD extract group(25 μg·mL-1),high dose SD extract group(50 μg·mL-1)and dexamethasone(DXMS)group(100 μg·mL-1)were set up,with intervention for 30 minutes.MTT test was used to detect the effect of low,medium,high-dose SD extract on activity of RBL-2H3 cell degranulation model.Toluidine blue staining was used to observe the morphology of degranulation cells and calculate degranulation rates.Immunofluorescence staining was used to detect expression of F-actin.Enzyme linked immunosorbent assay(ELISA)was used to detect the levels of β-aminohexosidase,histamine,interleukin-4(IL-4),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ).Western blot was used to detect the expressions of phosphatidylinositide-3 kinase(PI3K),phosphorylation-PI3K(p-PI3K),protein kinase B(AKT),phosphorylation-AKT(p-AKT),p38 mitogen activated protein kinase(p38MAPK),phosphorylation-p38MAPK(p-p38MAPK),nuclear factors-κB(NF-κB),phosphorylation-NF-κB(p-NF-κB),extracellular regulated kinases(ERK)and phosphorylation-ERK(p-ERK)protein.[Results]The low,medium,high doses of SD extract(5,25,50 μg·mL-1)had no significant effects on the activity of RBL-2H3 cells(P>0.05).Compared with blank control group,the number of toluidine blue stained cells of model group was decreased,cells shape rounded,degranulation rate was increased,expression of F-actin was decreased,the levels of β-aminohexosidase,histamine,IL-4,IL-6,TNF-α were increased,IFN-γ level was decreased,the expressions of p-PI3K/PI3K,p-AKT/AKT,p-p38MAPK/p38MAPK,p-NF-κB/NF-κB and p-ERK/ERK were increased(P<0.01).Compared with model group,the F-actin expression of low,medium,high doses of SD extract groups and DXMS group was increased,levels of β-aminohexosidase,histamine,IL-4,IL-6 and TNF-α were decreased,IFN-γ level was increased(P<0.01),the expressions of p-PI3K/PI3K,p-AKT/AKT,p-p38MAPK/p38MAPK,p-NF-κB/NF-κB and p-ERK/ERK were decreased(P<0.05,P<0.01);the number of toluidine blue stained cells in medium,high dose SD extract groups and DXMS group was increased with spindle cell shape,degranulation rate was decreased(P<0.01).Compared with low dose SD extract group,degranulation rate of high dose SD extract group and DXMS group was decreased(P<0.01),F-actin expression was increased(P<0.05),p-p38MAPK/p38MAPK expressions were decreased(P<0.01).[Conclusion]SD extract inhibited degranulation of IgE sensitized RBL-2H3 cell and decreased the levels of inflammatory mediators,its mechanism may be related to the inhibition the phosphorylation expression of PI3K/AKT,p38MAPK/NF-KB and ERK.
2.Serum metabolomics-based study on the mechanism of action of bergapten in the treatment of liver fibrosis
Huixing WU ; Zhenhua ZHANG ; Changrui LONG ; Guifen GUO ; Yanyu WANG ; Yanchun CHEN ; Juxiong FU ; Shijian XIANG ; Benjie ZHOU ; Chengyu LU
China Pharmacy 2024;35(13):1570-1575
OBJECTIVE To study the effects of bergapten in the treatment of liver fibrosis and its mechanism based on serum metabolomics. METHODS Forty mice were divided into normal control group (0.5% carboxymethyl cellulose sodium solution), model group (0.5% carboxymethyl cellulose sodium solution), and BP low-dose and high-dose groups (50, 100 mg/kg), with 10 mice in each group. Except for the normal control group, the other three groups were all treated with carbon tetrachloride to induce liver fibrosis model; they were given relevant medicine/solution intragastrically, once a day, for consecutive 8 weeks. After the last medication, the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected, and liver pathological changes were observed; the expressions of α-smooth muscle actin (α-SMA) and Collagen Ⅰ were detected in liver tissue; the serum of the mice was collected for metabolomics analysis. RESULTS Compared with the model group, serum levels of ALT and AST and protein expressions of α-SMA and Collagen Ⅰ in liver tissue were decreased significantly in BP high-dose and low-dose groups (P<0.05), while liver fibrosis was improved significantly. Meanwhile, metabolomics analyses showed that there were a total of 175 serum differential metabolites in the BP high-dose group and model group, of which 18 substances were upregulated and 157 substances were downregulated; the main metabolic pathways involved in bergapten intervention were pyrimidine metabolism, butanoate metabolism, fatty acid synthesis, tyrosine metabolism, β-alanine metabolism, nicotinic acid and nicotinamide metabolism, glutathione metabolism, etc. CONCLUSIONS BP is effective in the treatment of liver fibrosis by regulating pyrimidine metabolism, butanoate metabolism, glutathione metabolism and so on in rats with liver fibrosis.
3.Correlation between individualized coronary volumetric perivascular characterization index and coronary heart disease
Yanchun ZHANG ; Li CHENG ; Yue DAI ; Lixiang XIE ; Hao WANG ; Huan LIU ; Guangyi CHEN
Journal of Practical Radiology 2024;40(9):1444-1448
Objective To explore the correlation between coronary fat attenuation index of perivascular adipose tissue(FAIPVAT),as well as volumetric perivascular characterization index(VPCI),and coronary heart disease(CHD).Methods A total of 112 patients who underwent coronary computed tomography angiography(CCTA)and coronary angiography(CAG)examinations within 2 weeks were selected.The patients were divided into CHD group and control group according to the degree of coronary artery stenosis,and were divided into calcified plaque group,non-calcified plaque group and mixed plaque group according to the nature of plaque.The correlation between FAIPVAT,VPCI and CHD were evaluated.Results Of the 112 patients,71 patients in the CHD group and 41 patients in the control group.There were significant differences in gender,age,FAIPVAT and VPCI between the two groups.FAIPVAT and VPCI were positively correlated with CHD(r=0.445,P<0.01;r=0.669,P<0.01).FAIPVAT and VPCI were analyzed by receiver operating characteristic(ROC)curve,the area under the curve(AUC)of FAIPVAT was 0.770,the cut-off value was-81.659 HU,the sensitiv-ity was 0.592,and the specificity was 0.878,while the AUC of VPCI was 0.892,the cut-off value was 8.056,the sensitivity was 0.887,and the specificity was 0.805.There were significant differences in FAIPVAT and VPCI between non-calcified plaque group and calci-fied plaque group.FAIPVAT and VPCI of mixed plaque group and calcified plaque group were significantly different.Conclusion FAIPVAT and VPCI have some certain correlation with CHD.FAIPVAT and VPCI can accurately evaluate the risk level of CHD and coronary artery inflammation,and the value of individualized VPCI is higher.
4.Clinical effectiveness of dynamic joint mobilization combined with core stability training for nonspecific low back pain treatment
Lingling ZHU ; Yanchun CHEN ; Liang FANG ; Chengyan CAI ; Ting LIU
The Journal of Practical Medicine 2024;40(18):2602-2606
Objective To observe the clinical efficacy of dynamic joint mobilization combined with core stability training in the treatment of nonspecific low back pain.Methods 60 patients with nonspecific low back pain were randomly assigned into either the treatment group or control group.Each group had 30 patients.The treatment group received a therapeutic regimen combining dynamic joint mobilization and core stability training,while the control group only received core stability training.Both groups were evaluated for therapeutic effectiveness using the Visual Analogue Scale(VAS)for pain,the Oswestry Disability Index(ODI),and the range of motion(ROM)of the lumbar spine before treatment,and at the 1st,3rd,and 6th weeks after treatment.At the conclusion of the treatment,a thorough assessment of the overall therapeutic efficacy was performed.Results At the 1st,3rd,and 6th weeks post-treatment,both groups showed statistically significant differences in VAS scores,ODI scores,and ROM scores over time(P<0.05).The improvements in these indices were significantly greater in the treatment group compared to the control group(P<0.05).The treatment group had considerably higher therapeutic effective-ness compared to the control group(P<0.05).Conclusions Dynamic joint mobilization combined with core stabil-ity training is effective in treating nonspecific low back pain.It can help with pain relief,lumbar and back function restoration,and lumbar and back mobility improvement.This approach is worthy of clinical application and promotion.
5.Microbiome study of deep invasive endometriosis lesions
Ping HUANG ; Kangyun LAN ; Yanchun LIANG ; Qing CHEN ; Ying JIN ; Guangyuan CHEN ; Gang NIU
The Journal of Practical Medicine 2024;40(21):3023-3030
Objective To investigate the microbiome composition of deep invasive endometriosis lesions,offering novel insights into its pathogenesis,diagnosis,and treatment strategies.Methods From May 2021 to May 2022,we collected samples of normal endometrium(normal group,n=10),endometrium from patients with deep infiltrating endometriosis(DIE endometrium group,n=14),and lesions from these patients(DIE lesion group,n=10)for 16s rRNA sequencing analysis.We employed the bacterial community diversity algorithm(alpha diversity and beta diversity),principal coordinate analysis(PCoA)distance matrix algorithms(Bray-Curtis and Unifrac matrices),as well as the biological identification algorithm(LeFSe)to investigate microbiome differences between groups and identify differentially abundant bacteria and enriched KEGG functions.Results The diversity of the three groups did not show any significant difference,while the β diversity exhibited a statistically significant distinc-tion(P=0.005 5).The focus group had a relatively low abundance of Firmicutes at the phylum level and a relatively increased abundance of Fusobacteria.At the genus level,there was an increased relative abundance of Enterococcus and Prevotella.LEFse analysis suggested that Proteobacteria and Gammaproteobacteria were the main dominant in the lesion group.Notably,Gamma proteobacteria emerged as a key differential species among the three groups'microbial flora with statistical significance(P=0.021 2).Conclusion Compared to the normal group and DIE group,there was a significant increase in the abundance of Fusobacteria,Proteobacteria,Gammaproteobacteria,Enterococcus,and Prevotella.These findings offer novel therapeutic targets for the prevention and intervention of deep infiltrating endometriosis.
6.Caries experience and its correlation with caries activity of 4-year-old children in Miyun District of Beijing
Xinxin CHEN ; Zhe TANG ; Yanchun QIAO ; Wensheng RONG
Journal of Peking University(Health Sciences) 2024;56(5):833-838
Objective:To investigate the prevalence of dental caries of 4-year-old children in Miyun District of Beijing by international caries detection and assessment system(ICDAS),to detect the caries activity Cariostat value and to analyze the correlation between the Cariostat value and dental caries.Methods:Totally 815 children aged 4 years in 7 kindergartens in Miyun District of Beijing were recrui-ted.The clinical examination of all children was conducted by one examiner using ICDAS.The oral de-birs and plaques were collected by one doctor who recorded the Cariostat scores.The results of clinical examination were compared between genders.At the same time,the prevalence of dental caries,the mean d3-6ft/d3_6fs and d1-6ft/d1-6fs among high Cariostat scores group(2.0-3.0),medium Cariostat scores group(1.5)and low Cariostat scores group(0-1.0)were compared.The distributions of incipi-ent caries in different Cariostat scores groups were compared among children with incipient caries only.Results:All the children had incipient caries,and 78.3%of the children had cavitated caries with ICDAS score of 3 or above.The mean d1-2t scores were 9.76±3.65,the mean d3-6ft scores was 4.64±4.43 and the mean d1-6ft scores were 14.41±3.42.The incipient caries with ICDAS score of 1-2 were widely distributed,accounting for 67.7%of the total numbers of caries.There was no significant diffe-rence in caries prevalence and caries experience between genders(P>0.05).The proportion of children with high Cariostat scores in boys(43.6%)was higher than that in girls(33%)and the difference was statistically significant(P<0.05).With the increase of Cariostat scores,the prevalence of cavitated caries,the mean d3-6ft/d3-6fs and d1-6ft/d,_6fs scores in children was on the increase and the difference among the three groups was statistically significant(P<0.05).For children with incipient caries only,the distribution of incipient caries in different Cariostat scores groups was no significant difference(P>0.05).Conclusion:ICDAS can detect early enamel demineralization of deciduous teeth in children.The prevalence of dental caries among 815 4-year-old children in Miyun District of Beijing is more serious,and incipient caries is widely distributed in children.Cariostat value reflects the status of cavi-tated caries and has no correlation with the distribution of incipient caries.Therefore,the combined ap-plication of ICDAS and Cariostat caries activity detection method is helpful for the detection of incipient caries and screening of caries high-risk children,which has great significance for the comprehensive ma-nagement of caries in children and the formulation of early preventive measures.
7.Clinicopathological diagnosis and treatment of undifferentiated(embryonal) sarcoma of the liver
Wei GUO ; Qing HU ; Kun WANG ; Zhihong CHEN ; Rongguang WEI ; Hong YANG ; Yanchun LI
Chinese Journal of General Surgery 2023;38(1):7-11
Objective:To investigate the clinicopathological features, differential diagnosis, treatment and prognosis of undifferentiated embryonal sarcoma of the liver (UESL).Methods:Five UESL cases operated on at Hunan Provincial People's Hospital from 2014 to 2021 were retrospectively analyzed. H&E and immunohistochemical staining were done for pathological observation.Results:The 5 UESL patients(two boys,three girls) were 0.5 to 15 years old, all underwent radical surgical resection. In 3 cases tumors located in right liver, 1 in left liver, 1 in both lobes. Radiographically and visually, the tumor is a large cystic solid mass, microscopically composed of myxoid stroma and undifferentiated stromal cells, with pleomorphic tumor giant cells and characteristic eosinophilic bodies. All 5 patients are now alive after surgical resection: 1 patient achieved disease-free survival of more than 91 months after surgery alone. Two patients had recurrence after surgery and received surgical resection plus chemotherapy or chemotherapy alone. They achieved survival of more than 35 and 16 months, respectively. Two patients were treated with chemotherapy or chemotherapy plus radiotherapy after surgery and survived more than 49 and 31 months without recurrence, respectively.Conclusions:UESL is a rare and highly malignant mesenchymal tumor with characteristic pathologic morphology. Radical resection is the key to the treatment for UESL, and chemotherapy and radiotherapy should be carried out after surgery.
8.Clinical genetic analysis of an infant with 1q21.3-qter duplication and Xpter-p11 deletion caused by maternal balanced chromosomal translocation.
Lin LIN ; Chen ZHAO ; Yanchun LV ; Xiangyu ZHAO ; Lin LI
Chinese Journal of Medical Genetics 2022;39(5):514-517
OBJECTIVE:
To investigate the clinical phenotype and genetic diagnosis of an infant featuring multiple hair and hyperbilirubinemia.
METHODS:
Conventional G-banding analysis, chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH) for the patient were conducted, G-banding analyses of peripheral blood for the infant's parents were also performed.
RESULTS:
We investigated an infant who carries a unbalanced, maternally inherited karyotype 46, X, der (X) t (X;1) (p11.22; q21.3) in which CMA and FISH analyses disclosed a 1q21.3q44 duplication of 93.03 Mb and Xp22.33p11.22 deletion of 54.53 Mb.
CONCLUSION
The phenotypes of this infant can probably be attributed to the 1q21.3q44 duplication and Xp22.33p11.22 deletion, which were maternally inherited.
Chromosome Banding
;
Chromosome Deletion
;
Genetic Testing
;
Humans
;
In Situ Hybridization, Fluorescence
;
Karyotyping
;
Translocation, Genetic
9.Evaluation on measurement uncertainty of correction factors of fluconazole impurities determined by HPLC standard curve method
Ting XIAO ; Bufang MA ; Chen WANG ; Shangchen YAO ; Yanchun FENG ; Baoming NING
Journal of China Pharmaceutical University 2022;53(3):306-313
In this paper, the uncertainties of correction factors of fluconazole impurities determined by HPLC standard curve method were evaluated, and the main common factors affecting the accuracy of standard curve method were found, so as to improve the accuracy of the method.In this study, the corresponding fitting lines of fluconazole and its impurities A, B, C, D, F and I were established respectively, and the ratio of the slope of fitting lines of each impurity and its corresponding principal component was calculated as the correction factor of the impurity.Then on the basis of GUM method, the uncertainty of each impurity correction factor determined by standard curve method was evaluated according to the established uncertainty evaluation scheme of correction factor determination process.The correction factor and uncertainty of fluconazole impurities A, B, C, D, F and I were 1.068 ± 0.046, 0.102 ± 0.005, 0.0582 ± 0.0031, 1.382 ± 0.121, 0.802 ± 0.067 and 1.383 ± 0.119, respectively, and the coverage factor k was 2.Finally, the contribution rate of each uncertainty component was calculated.In the relative combined standard uncertainties urel(f) of fluconazole impurities A, B, C, D, F and I correction factors, the sum of contribution rate of slope uncertainty urel(K) of the linear equation of principal component and its impurity is more than 85%; in the slope uncertainties urel(K) of linear equation, the contribution rates of uncertainties of solution concentration in 8 of 12 data groups are more than 80%, and the contribution rates of uncertainties introduced by reference substance content in solution concentration are about 80%.It can be seen that the preparation of linear solution concentration is the most influential factor in the determination of impurity correction factor by standard curve method, followed by the linear fitting process.In the preparation process of linear solution concentration, the purity of reference substance is the most influential factor, followed by weighing and pipetting times.The conclusion can help the experimenters to better formulate experimental plans and ensure the accuracy of the results when doing similar work.
10.Study on regulatory mechanism of compatibility of ginseng and gecko dispensing granule on neuroendocrine immune network in model rats with kidney yang deficiency
Bei ZHOU ; Qiaofeng ZHU ; Yudan CHEN ; Liting FENG ; Shuling LIU ; Yanchun WU
China Pharmacy 2022;33(11):1313-1319
OBJECTIVE To explore the regulatory mechanism of compatibility of ginseng and gecko dispensing granule on kidney yang deficiency model rats. METHODS Male SD rats were randomly divided into normal group (no modeling ,no administration),model group (modeling,no administration ),Jinkui shenqi pill group (modeling,dose of 2.33 g/kg),ginseng group(modeling,dose of 0.53 g/kg),gecko group (modeling,dose of 0.21 g/kg)and compatibility group (modeling,ginseng 0.53 g/kg and gecko 0.21 g/kg). The body mass and anal temperature of rats were measured at different time points ;the serum levels of cAMP ,cGMP,CRH,ACTH,CORT,T,T3,T4,E2,IgG and IgM were measured ;the pathomorphological changes of adrenal gland ,thyroid gland and testis were observed ;mRNA expression of CRH ,thyroid stimulating hormone releasing hormone (TRH)and gonadotropin releasing hormone (GnRH)in hypothalamus were detected. RESULTS Compared with model group ,the anal temperature ,the levels of cAMP ,CRH,ACTH,CORT,T3,T and cAMP/cGMP ,T/E2 in serum and mRNA expressions of TRH and GnRH in hypothalamus were significantly increased in the compatibility group (P<0.05 or P<0.01);the levels of cGMP,E2 and IgG in serum and mRNA expression of CRH in hypothalamus decreased significantly (P<0.05 or P<0.01); the pathological injuries of adrenal gland ,thyroid gland and testis were all improved. Compared with ginseng or gecko dispensing granules alone ,the anal temperature and T/E 2 of rats in the compatibility group increased significantly ,and mRNA expression of CRH in hypothalamus decreasedsignificantly (P<0.05 or P<0.01). CONCLUSIONS Thecompatibility of ginseng and gecko dispensing granule has a synergistic regulatory effect on kidney yang deficiency model rats , the mechanism of which may be associated with hypothalamus-pituitary-adrenal axis , hypothalamus-pituitary-thyroid axis , hypothalamus-pituitary-gonad axis and neuroendocrine immune network formed by immune function. Compatible drugs are better than single drugs.

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