1.Improvement of Colonic Mucosa Inflammatory Response in Mice with Ulcerative Colitis by Xiezhuo Jiedu Recipe Through miRNA-155-5p/JAK2/STAT3 Pathway
Chaodi SUN ; Mengmeng ZHAO ; Xiaomeng LANG ; Jie REN ; Xin KANG ; Jiancong CUI ; Sujie JIA ; Yujing MA ; Yue LIU ; Qiang CHUAI ; Wenjing ZHAI ; Jianping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):174-182
ObjectiveThe differential expression of microRNAs (miRNAs) between the active stage and the remission stage of ulcerative colitis (UC) was analyzed by bioinformatics method, and the regulatory relationship was constructed by screening the differentially expressed genes (DEGs). The mechanism of Xizhuo Jiedu recipe in the treatment of UC was speculated and verified by animal experiments. MethodThe miRNAs data set of colonic mucosa tissue of UC patients was obtained from the gene expression database (GEO), and the most differentially expressed miRNAs were screened by GEO2R, Excel, and other tools as research objects. TargetScan, miRTarbase, miRDB, STRING, TRRUST, and Matescape databases were used to screen key DEGs, predict downstream transcription factors (TFs), gene ontology (GO), and conduct Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The key signaling pathways were selected for animal experiments. In animal experiments, the UC mouse model was prepared by making the mouse freely drink 2.5% dextran sodium sulfate (DSS). Xiezhu Jiedu recipe and mesalazine were given by gavage for seven days, and the inflammatory infiltration of colonic mucosa was observed by hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of miR-155-5p in colon tissue. Immunohistochemistry and Western blot were used to detect the protein expression levels of cytokine signal transduction inhibitor (SOCS1), phosphorylated transcriptional signal transductor and activator 3 (p-STAT3), phosphorylated Janus kinase 2 (p-JAK2), and retinoic acid-associated orphan receptor-γt (ROR-γt). The expression levels of transforming growth factor-β (TGF-β), interleukin-17 (IL-17), interleukin-6 (IL-6), and interleukin-10 (IL-10) in serum were detected by enzyme linked immunosorbent assay (ELISA). ResultThe GSE48957 dataset was screened from the GEO database, and miR-155-5p was selected as the research object from the samples in the active and remission stages. 131 DEGs were screened. The GO/KEGG enrichment analysis was closely related to biological processes such as positive regulation of miRNA transcription and protein phosphorylation, as well as signaling pathways such as stem cell signaling pathway, IL-17 signaling pathway, and helper T cell 17 (Th17) cell differentiation. The Matescape database was used to screen out 10 key DEGs, among which SOCS1 was one of the key DEGs of miR-155-5p. Further screening of the TFS of key DEGs revealed that STAT3 was one of the main TFs of SOCS1. The results of animal experiments showed that Xiezhu Jiedu Recipe could effectively down-regulate the mRNA expression of miR-155-5p and protein expression of p-STAT3, p-JAK2, and ROR-γt in colon tissue of UC mice and the expression of IL-17 and IL-6 in serum of UC mice, up-regulate the protein expression of SOCS1 and the expression of TGF-β and IL-10, increase the level of anti-inflammatory factors, and reduce inflammatory cell infiltration. ConclusionIt is speculated that Xizhuo Jiedu recipe may interfere with SOCS1 by regulating the expression of miR-155-5p in UC mice, inhibit the phosphorylation of STAT3, inhibit the differentiation of CD4+ T cells into Th17 cells, reduce the levels of pro-inflammatory factors (IL-17 and IL-6), and increase the levels of anti-inflammatory factors (TGF-β and IL-10). As a result, the inflammation of colon mucosa in UC mice was alleviated.
2.The influence of spectral CT monochromatic imaging technique on image quality and liver volume measurement in CT portal venography
Yong FAN ; Yijun LIU ; Beibei LI ; Shigeng WANG ; Wei WEI ; Xiaoyu TONG ; Zhiming MA ; Anliang CHEN ; Yujing ZHOU
Journal of Practical Radiology 2024;40(9):1536-1540
Objective To investigate the influence of spectral CT monochromatic imaging technique on image quality and liver vol-ume measurement in computed tomography portal venography(CTPV).Methods A total of 120 patients who underwent contrast-enhanced abdominal CT examination were prospectively selected and randomly divided into group A and group B.The group A(n=60)was scanned with conventional parameters such as 120 kVp,Smart mA mode,and image reconstruction of 60%adaptive statistical iterative reconstruction-Veo(ASIR-V);while the group B(n=60)was with instantaneous switching 80/140 kVp,gemstone spectral imaging(GSI)Assist mode,and image reconstruction of 60%ASIR-V.In group B,six subgroups of images from 75 to 50 keV(with 5 keV interval)were recorded as subgroups B1 to B6.On the axial images,the CT values and standard deviation(SD)values of por-tal vein,liver and erector spinae were measured,and then the contrast-to-noise ratio(CNR)of portal vein and liver were calculated.The Liver Segmentation software was used to segment the liver in groups A and B,the liver volumes by automatic segmentation and manual segmentation(golden standard)were recorded,and then the volume difference rate was calculated.The overall image quality and automatic liver segmentation results were evaluated by two radiologists using a 5-point scale.Results In terms of overall image quality,subgroup B6 achieved the highest score and was superior to group A(P<0.001).In terms of liver segmentation,subgroup B3 had the highest score and was superior to group A(P<0.001).With the decrease of keV,the CT values,SD values and CNR of portal vein and liver in group B were gradually increased(P<0.05),in which subgroup B6 was higher than that in group A(P<0.001).The volume difference rate initially decreased and then increased with the decrease of keV.Except for subgroups B2 and B3,the differences were statistically significant between other subgroups and group A(P<O.001),and the subgroup B3 had the lowest volume difference rate.Conclusion Spectral CT monochromatic ima-ging technique has an influence on CTPV image quality and liver volume measurement.The 50 keV images are the best for displaying portal vein,and the automatic liver segmentation volume of 65 keV images is closest to the real liver volume.
3.Detection of 13 Paralytic shellfish toxins in human whole blood by high-performance liquid chromatography-tandem mass spectrometry
Xiuqin MA ; Qiang JIE ; Yujing LUAN ; Zeyu WANG ; Fanglin WANG
Chinese Journal of Forensic Medicine 2024;39(4):458-461,466
Objective To establish the analysis method of 13 Paralytic shellfish toxins(PSTs)in human whole blood by high-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Methods 13 kinds of PSTs were extracted from the whole blood using acetonitrile with 1%acetate(1︰3,v︰v)solution,and separated by UPLC BEH Amide chromatographic column(100mm×2.1mm,1.7μm).The samples were detected by an electrospray ionization source(ESI)and positive and negative ion multiple reaction monitoring(MRM)mode,and quantified by matrix matching curve external standard method.Results The results showed that 13 kinds of PSTs in blood samples were linear well in the ranges of 1~100 ng/mL,with the correlation coefficient(r2)>0.995.The limit of detection(LODs)of the method were 0.5~2 ng/mL,the limit of quantitation(LOQs)were 1~4 ng/mL,and the recoveries of the method were 65.55%~114.12%.Conclusion The method is highly sensitive,reproducible,and can qualify and quantify thirteen toxins at the same time,which is suitable for the rapid detection of PSTs in whole blood samples.
4.Seroepidemiology analysis of Epstein-Barr virus infection in children in Shenyang from 2022 to 2023
Jingxuan SUN ; Mingyi XU ; Zhongyang LIU ; Ying QI ; Yanping MA ; Qiang RUAN ; Yujing HUANG
Journal of China Medical University 2024;53(10):865-869
Objective To analyze the seroepidemiology characteristics of Epstein-Barr virus(EBV)infection in children in Shenyang.Methods From June 2022 to May 2023,serum was collected from 12 083 children at Shengjing hospital of China Medical University.Serum capsid antigen(VCA)-IgM,VCA-IgG,EBV nuclear antigen-IgG(EBNA-IgG),and early antigen-IgG(EA-IgG)antibodies were detected using the LIAISION chemiluminescence immunoassay.EBV-DNA was detected using real-time PCR.Differences in antibody positivity rates between sexes,ages,and seasons of onset were compared.Results In 12 083 patients,the positive rates of VCA-IgM,VCA-IgG,EBNA-IgG,and EA-IgG were 9.95%(1 202/12 083),50.57%(6 110/12 083),46.03%(5 562/12 083)and 4.93%(596/12 083).The positive rates of VCA-IgM and EA-IgG were lower in male patients(9.09%and 4.44%,respectively)than in female patients(11.10%,5.60%;all P<0.05).The differences in the positive rates for VCA-IgM,VCA-IgG,EBNA-IgG,and EA-IgG antibodies in children of different ages were statistically significant(all P<0.05).Differences in the positive rates of VCA-IgG,EBNA-IgG,and EA-IgG antibodies were statistically significant when compared between seasons(all P<0.05).Fourteen EBV antibody-positive combinations were detected,of which the main combination was VCA-IgG and EBNA-IgG double-positive,with a total of 4 741 cases(39.24%).Of the 3 712 children who underwent EBV-DNA detection testing,3 034(81.73%)were EBV-DNA-negative and 678(18.27%)were EBV-DNA-posi-tive.VCA-IgG and EBNA-IgG double positivity was the most common in EBV-DNA-negative and EBV-DNA-positive children,accounting for 983(26.48%)and 194 cases(5.23%),respectively.Conclusion Both VCA-IgG and EBNA-IgG antibodies are main positive in children with EBV infection in Shenyang.The positive rate of EBV antibodies is lower in boys than in girls.The positive rates of EBV anti-bodies differ in children of different ages and seasons of onset.
5.Study on the mechanism of Tianshui Dichang Decoction in the treatment of ulcerative colitis based on network pharmacology and experimental verification
Peitong LI ; Xiaomeng LIU ; Yujing MA ; Ziwei DONG ; Sen YANG ; Qian YANG ; Jianping LIU ; Xiaomeng LANG
International Journal of Traditional Chinese Medicine 2023;45(11):1399-1407
Objective:To explore the potential mechanism of Tianshui Dichang Decoction in the treatment of ulcerative colitis through network pharmacology and experimental verification.Methods:The active components and targets of Tianshui Dichang Decoction were screened by TCMSP. The related targets of ulcerative colitis were screened by OMIM, GeneCard and TTD databases, and the effective component targets of Tianshui Dichang Decoction were intersected with the potential targets of ulcerative colitis. The PPI network was constructed by STRING database to screen the core targets, and the "Chinese materia medica-disease-active components-target" network was constructed by Cytoscape 3.8.0 software. GO function and KEGG pathway enrichment analysis were carried out using Metascape database. 48 mice were divided into control group, model group, mesalazine group (0.3 g/kg) and Tianshui Dichang Decoction low-, medium-, and high-dosage groups (7.5,15 and 30 g/kg) according to random number table method, with 8 mice in each group. Except the control group, the ulcerative colitis model was established in other groups. After 7 days of intervention with corresponding drugs, the disease activity index (DAI) was scored, the pathological changes of colon were observed by HE staining, and the expressions of IL-6, STAT3mRNA and protein in colon tissue were detected by PCR and Western blot methods.Results:Totally 127 active components in Tianshui Dichang Decoction and 560 targets of ulcerative colitis were obtained. 89 intersecting targets of Tianshui Dichang Decoction and ulcerative colitis were obtained, and the core targets included IL6, TNF, IL1B, AKT1, TP53, VEGFA, JUN, PTGS2, CXCL8, CCL2, STAT3, MMP9 and so on. Oxidative stress response, lipopolysaccharide metabolism, bacterial response, signal transduction and other biological processes were mainly involved, mainly through the cancer pathway, IL17, TNF, MAPK and other signal pathways to play a role in the treatment of ulcerative colitis. The results of experimental verification showed that the DAI score, the expressions of IL-6 and STAT3 protein in colon tissue of Tianshui Dichang Decoction medium- and high-dosage groups.decreased ( P<0.05). The levels of IL-6 and STAT3 mRNA in colon tissue decreased in the Tianshui Dichang Decoction low-, medium- and high-dosage groups.groups ( P<0.05). Conclusion:Tianshui Dichang Decoction has a certain therapeutic effect on UC through component-multitarget-signal pathway, and its mechanism is related to regulating IL-6/STAT3 signal pathway and inhibiting intestinal mucosal inflammation.
6.Research advances on conductive hydrogels and their applications in the repair of diabetic wounds
Yang LYU ; Wanli CHU ; Yu CHEN ; Yujing LI ; Xilan MA
Chinese Journal of Burns 2023;39(6):586-590
As a new type of functional wound dressing, conductive hydrogel, shows broad prospects of application in the field of wound repair due to its suitable electrical conductivity, good moisture retention, excellent biocompatibility, and biological effects such as mediating cell migration and proliferation, and promoting angiogenesis and collagen deposition. Combined with the clinical electrical stimulation therapy, the conductive hydrogel primarily showed curative effects of promoting granulation tissue formation, re-epithelialization, and wound healing, providing a new treatment idea for the repair of diabetic wounds. This review summarized the research advances of electronic conductive hydrogels and ionic conductive hydrogels in recent years based on different conductive mechanisms. Meanwhile, the applications of conductive hydrogel in the diabetic wound repair were specifically introduced, and the future development of conductive hydrogel wound dressing was prospected.
7.Clinical characteristics of human cytomegalovirus pneumonia in children and the diagnosis value of detecting human cytomegalovirus DNA in bronchoalveolar lavage fluid
Xiangjing MENG ; Mengqing LU ; Yang LI ; Yujing HUANG ; Ying QI ; Qiang RUAN ; Yanping MA
Chinese Pediatric Emergency Medicine 2022;29(4):271-276
Objective:To investigate the clinical features, therapy and prognosis of human cytomegalovirus(HCMV)pneumonia in pediatric patients, and to analyze the diagnosis value of detecting HCMV DNA in bronchoalveolar lavage fluid(BALF)by real-time PCR.Methods:The clinical characteristics of 58 pediatric inpatients who were HCMV DNA positive in BALF were retrospectively reviewed.All the patients were from Shengjing Hospital of China Medical University from January 2015 to December 2019.Clinical, radiologic, laboratory and microbiologic data was collected for each patient.The study cohort was divided into HCMV productive infection and latent infection consisting of 22 and 36 patients respectively, based on the HCMV active infection in lung or not.Receiver operating characteristic(ROC)curve was used to assess utility of detecting HCMV DNA in BALF and establish a threshold for diagnosis.Results:(1)Compared with patients in latent infection group, the children in productive infection group had a lower age of onset( P<0.05), a higher proportion of male( P<0.05), and more prolonged hospitalization stay( P<0.05). Pulmonary rales, hypoxemia and higher AST, CK, LDH in serum were easier to detect in productive infection group( P<0.05). Higher HCMV DNA copies in BALF was also detected( P<0.01). Patients in productive infection group had significantly more exposure to additional oxygen treatment or mechanical ventilation and systemic hormone therapy( P<0.05), while with poorer outcomes( P<0.05). (2) ROC curve analysis showed that the AUC for HCMV DNA in BALF in diagnosis of HCMV pneumonia was 0.708 with a threshold of 8.83×10 3 copies/mL, a sensitivity of 77.27%, and a specificity of 58.33%. Conclusion:Those who are diagnosed HCMV pneumonia have a lower age of onset with higher male proportion.These children suffered severer clinical signs.The patients with HCMV DNA copies higher than 8.83×10 3 copies/mL in BALF would be more likely to be diagnosed as HCMV pneumonia.
8.Local recurrence pattern of pT 1-2N 1 breast cancer after modified radical mastectomy—a pooled-analysis of 5442 patients from 12 centers
Xinyuan GUO ; Yujing ZHANG ; Na ZHANG ; Yu TANG ; Xuran ZHAO ; Hao JING ; Hui FANG ; Ge WEN ; Jing CHENG ; Mei SHI ; Qishuai GUO ; Hongfen WU ; Xiaohu WANG ; Changying MA ; Yexiong LI ; Hongmei WANG ; Min LIU ; Shulian WANG
Chinese Journal of Radiation Oncology 2022;31(3):248-252
Objective:To analyze locoregional recurrence (LRR) pattern of patients with pT 1-2N 1 breast cancer after modified radical mastectomy, with and without adjuvant radiotherapy (RT). Methods:A total of 5442 eligible patients with breast cancer from 12 Chinese centers were included. The LRR sites and the effect of RT at different sites on recurrence in patients with and without RT were analyzed. The Kaplan-Meier method was used to calculate the cumulative LRR rate, and the difference was compared by the log-rank test.Results:With a median follow-up time of 63.8 months for the entire cohort, 395 patients developed LRR. The chest wall and supraclavicular fossa were the most common LRR sites, regardless of RT or molecular subtypes. The 5-year chest wall recurrence rates for patients with and without chest wall irradiation were 2.5% and 3.8%( P=0.003); the 5-year supraclavicular lymph nodal recurrence rates for patients with and without supraclavicular fossa irradiation were 1.3% and 4.1%( P<0.001); the 5-year axillary recurrence-free rates for patients with and without axillary irradiation were 0.8% and 1.5%( HR=0.31, 95% CI: 0.04-2.23, P=0.219); and the 5-year internal mammary nodal recurrence-free rates for patients with and without internal mammary nodal irradiation were 0.8% and 1.5%( HR=0.45, 95% CI: 0.11-1.90, P=0.268). Conclusions:The chest wall and supraclavicular fossa are the most common LRR sites of patients with pT 1-2N 1 breast cancer after modified radical mastectomy, which is not affected by adjuvant RT or molecular subtypes. The chest wall and supraclavicular fossa irradiation significantly reduce the risk of recurrence in the corresponding area. However, axillary and internal mammary nodal irradiation has no impact on the risk of recurrence in the corresponding area.
9.Cooperation-based sperm clusters mediate sperm oviduct entry and fertilization.
Yongcun QU ; Qi CHEN ; Shanshan GUO ; Chiyuan MA ; Yonggang LU ; Junchao SHI ; Shichao LIU ; Tong ZHOU ; Taichi NODA ; Jingjing QIAN ; Liwen ZHANG ; Xili ZHU ; Xiaohua LEI ; Yujing CAO ; Wei LI ; Wei LI ; Nicolas PLACHTA ; Martin M MATZUK ; Masahito IKAWA ; Enkui DUAN ; Ying ZHANG ; Hongmei WANG
Protein & Cell 2021;12(10):810-817
Animals
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Antigens, Surface/genetics*
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Cell Communication/genetics*
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Copulation/physiology*
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Fallopian Tubes/metabolism*
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Female
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Fertilization/genetics*
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GPI-Linked Proteins/genetics*
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Gene Expression Regulation
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Genes, Reporter
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Green Fluorescent Proteins/metabolism*
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Litter Size
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Luminescent Proteins/metabolism*
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Male
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Mitochondria/metabolism*
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Reproduction/genetics*
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Signal Transduction
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Sperm Count
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Sperm Motility/genetics*
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Spermatozoa/metabolism*
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Uterus/metabolism*
10.Study on in vitro and in vivo Genotoxicity of Emodin- 8-O-β-D-glucoside
Hairuo WEN ; Yujing YAN ; Jie SONG ; Rui AO ; Shuangcheng MA ; Qi WANG
China Pharmacy 2020;31(1):18-23
ABSTRACT OBJECTIVE:To evaluate the in vitro and in vivo genotoxicity of emodin-8-O-β-D-glucoside(EG),and to compare the difference of in vitro cell test and in vivo test of rats. METHODS:2D and 3D hepatocyte models were established by in vitro two-dimensional(2D)and three-dimensional(3D)cell culture. After modeling,2D and 3D hepatocyte were divided into blank control group(0.5% DMSO),mitomycin C group(positive control,0.1 μg/mL),EG low-dose,medium-dose and high-dose groups(10,50,200 μg/mL),respectively. The micronucleus ratio and tail DNA% of HepaRG cells were detected. SD rats were divided into blank control group(0.5% sodium carboxymethyl cellulose),ethyl methanesulfonate group(positive control,200 mg/kg),EG low-dose,medium-dose and high-dose groups(100,300,1 000 mg/kg),with 6 rats in each group. They were given medicine intragastrically for consecutive 15 d,once a day. 15 days later,the micronucleus formation rate of bone marrow polychromatic erythrocytes and hepatocytes,the tail DNA% and tail distance of peripheral blood lymphocytes and hepatocytes were measured. RESULTS:In the in vitro 2D HepaRG hepatocyte model,compared with blank control group,the micronucleus formation rate and tail DNA% of HepaRG cell were increased significantly in mitomycin C group (P<0.01). There was no statistical significance in micronucleus formation rate and tail DNA% of HepaRG cell among EG groups(P>0.05). In 3D HepaRG cell model, compared with blank control group, micronucleus formation rate and tail DNA% of HepaRG cell were increased significantly in mitomycin C group (P<0.01 or P<0.001), while tail DNA% of HepaRG cell wasincreased significantly in EG high-dose group(P<0.01). In the in vivo test,compared with blank control group,the micronucleus formation rate of bone marrow polychromatic erythrocytes and hepatocytes,the tail DNA% and tail distance of peripheral blood lymphocytes and hepatocytes were all increased significantly in ethyl methanesulfonate group(P<0.01). Tail DNA% of peripheral blood lymphocytes was increased significantly in EG high-dose group (P<0.01). There was no statistical significance in the micronucleus formation rate of bone marrow polychromatic erythrocytes and hepatocytes,the tail DNA% and tail distance of hepatocytes among EG groups(P>0.05);with the increase of dose,there was an increasing trend. CONCLUSIONS:The results of this study suggest that in 2D cell model,EG not lead to chromosome breakage and DNA damage,but the long-term administration and repeated administration in vivo of 3D cell model show that EG has a certain risk of DNA damage,so the evaluation results of 3D HepaRG cell model are more similar to those of rats in vivo. KEYWORDS Emodin-8-O-β-D-glucoside;Genotoxicity;Two-dimensional culture;Three-dimensional culture;Rat;Micronucleus test

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