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.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.
3.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.
4.Eligibility of C-BIOPRED severe asthma cohort for type-2 biologic therapies.
Zhenan DENG ; Meiling JIN ; Changxing OU ; Wei JIANG ; Jianping ZHAO ; Xiaoxia LIU ; Shenghua SUN ; Huaping TANG ; Bei HE ; Shaoxi CAI ; Ping CHEN ; Penghui WU ; Yujing LIU ; Jian KANG ; Yunhui ZHANG ; Mao HUANG ; Jinfu XU ; Kewu HUANG ; Qiang LI ; Xiangyan ZHANG ; Xiuhua FU ; Changzheng WANG ; Huahao SHEN ; Lei ZHU ; Guochao SHI ; Zhongmin QIU ; Zhongguang WEN ; Xiaoyang WEI ; Wei GU ; Chunhua WEI ; Guangfa WANG ; Ping CHEN ; Lixin XIE ; Jiangtao LIN ; Yuling TANG ; Zhihai HAN ; Kian Fan CHUNG ; Qingling ZHANG ; Nanshan ZHONG
Chinese Medical Journal 2023;136(2):230-232
5.Introduction to revision of Technical Specification for Occupational Health Surveillance
Chen YU ; Dehong LI ; Daoyuan SUN ; Zubing WANG ; Chaoqiang JIANG ; Xunmiao ZHANG ; Yongjian YAN ; Weiming YUAN ; Yiqun XUAN ; Xin QIAO ; Yujing XIA ; Qiuhong ZHU ; Qiang HOU ; Hong WANG ; Yiwen JIANG ; Xuetao ZHANG ; Fang QI ; Xiangpei LÜ ; Huanqiang WANG
China Occupational Medicine 2023;50(2):209-216
To revise GBZ 188 Technical Specification for Occupational Health Surveillance based on national laws, regulations, standards, specifications and legal documents of occupational disease, and combination with the actual situation in China. The main modifications are as follows: the occupational health surveillance for workers exposed to toluene (xylene may implement by reference), bromopropane, methyl iodide, ethylene oxide, chloroacetic acid, indium and its compounds, coal tar, coal tarasphalt, asphalt, β-naphthylamine, dust of metal and its compounds(tin, iron, antimony, barium and its compounds), hard metal dust, erionite dust, low temperature, laser, tick-borne encephalitis virus, Borrelia burgdorferi, and human immunodeficiency virus, for scraper or grind operators, and underground workers using squatting or kneeling position, crawling position, side-lying position, or shoulder position for a long period of time are included. The emergency health screening for workers exposed to arsenic, fluorine and its inorganic compounds, and acrylamide are included. The occupational medical examination (OME) for workers exposed to amino and nitro compounds of benzene, phosgene, monomethylamine, organic fluorine and dimethyl sulfate has been adjusted and made mandatory, with corresponding assessments required upon leaving the job. The special occupational health surveillance for workers exposed to mycobacterium tuberculosis and hepatitis virus is removed. The OME conclusion of reexamination is removed, and standardize recheck/additional inspection requirements. The optional items in OME performed before, during and after leaving post are removed, but the optional items in emergency medical examination are retained. Additional OME items are added. The Guideline for OME Summary Reports is added as informative appendix, and so on. The revised GBZ 188 Technical Specification for Occupational Health Surveillance is more scientific and practical.
6.Study on the Dynamic Change of Main Alkaloids Content in Aconitum Sinomontanum Nakai Processing Based on the Simulated Processing Technology
JIA Chunyan ; QIANG Yujing ; YANG Xicang ; MIAO Xiaolou ; LI Yun
Chinese Journal of Modern Applied Pharmacy 2023;40(15):2041-2047
OBJECTIVE To explore the dynamic changes of main alkaloids in the processing process of Aconitum sinomontanum Nakai prepared by liquorice juice. METHODS Aconitum sinomontanum Nakai with or without liquorice juice were steaming for 0, 0.5, 1, 2, 3, 4, 5 h, respectively, and then dried. HPLC was used to determine the content of lappaconitine, ranaconitine and new components in different processed products of Aconitum sinomontanum Nakai. The simulated processing was carried out with the reference substance of lappaconitine and ranaconitine. The new components were separated by preparative liquid and analyzed by NMR and UPLC-QTOF-MS. RESULTS With the extension of steaming time, the content of lappaconitine and ranaconitine decreased significantly during the steaming Aconitum sinomontanum Nakai with or without liquorice juice, and two new peaks(peak 1 and peak 2) were produced and their content increased observably, this indicated that the new compounds were not related to licorice juice, only related to steaming process. Compared with the steaming process without liquorice juice, the contents of toxic and effective components lappaconitine and ranaconitine decreased moderately steamed with liquorice juice, indicating that the addition of liquorice juice steaming was more reasonable; The results of the simulated processing showed that the contents of lappaconitine and ranaconitine gradually decreased in the processing. After isolation and identification find the compound 1(peak 2) was 5-hydroxymethylfurfural. CONCLUSION The main alkaloids components of Aconitum sinomontanum Nakai are changed greatly after processing, which may be closely related to the “reduction of toxicity and increase of efficiency” in the steaming process of adding liquorice juice.
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.Molecular mechanism underlying the inhibitory effect of propofol on lipopolysaccharide-induced pyroptosis of mouse bone marrow-derived macrophages.
Xuexia JI ; Yuanbo GUO ; Qianqi QIU ; Zhipeng WANG ; Yan WANG ; Jinquan JI ; Qiang SUN ; Yujing CAI ; Guobin ZHOU
Journal of Southern Medical University 2020;40(4):525-530
OBJECTIVE:
To investigate the molecular mechanism underlying the inhibitory effect of propofol on pyroptosis of macrophages.
METHODS:
Macrophages derived from bone marrow were extracted and divided into three groups: control group, LPS+ATP group and propofol+LPS+ATP group. The control group was not given any treatment; LPS+ATP group was given LPS 1 μg/mL stimulation for 4 h, then ATP 4 mM stimulation for 1 h; Propofol+LPS+ATP group was given propofol+LPS 1 μg/mL stimulation for 4 h, then ATP stimulation for 1 h. After treatment, the supernatant and cells of cell culture were collected. the cell activity was detected by CCK8 and flow cytometry. The inflammatory cytokines IL-1βand IL-18 were detected by Elisa. Western blot was used to detect the expression of caspase-1 protein and TLR4 on cell membran Immunohistochemical fluorescence was used to detect apoptosis of cells.
RESULTS:
LPS+ATP significantly decreased the viability of the macrophages and increased the cellular production of IL-1β and IL-18, activation of caspase-1 protein and the expression of TLR-4 on the cell membrane ( < 0.05). Treatment with propofol obviously reversed the changes induced by LPS+ATP.
CONCLUSIONS
LPS+ATP can induce pyroptosis of mouse bone marrow-derived macrophages, and propofol effectively inhibits such cell death, suggesting that propofol anesthesia is beneficial during operation and helps to regulate the immune function of in patients with sepsis.
Animals
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Caspase 1
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Lipopolysaccharides
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Macrophages
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Mice
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Propofol
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Pyroptosis
9. Analysis of human papillomavirus infection characteristics and genotypes in Shenyang area of Liaoning province
Te QI ; Yujing HUANG ; Chao LIU ; Bo WANG ; Hao WU ; Ye MIAO ; Qiang RUAN
Chinese Journal of Experimental and Clinical Virology 2019;33(4):389-393
Objective:
To investigate the distribution of human papillomavirus (HPV) infection characteristics and genotypes in Shenyang area of Liaoning province.
Methods:
HPV genes were detected in cervical exfoliated cells from 55, 548 patients by amplification and diversion hybridization.
Results:
A total of 9, 566 patients were positive for HPV infection with a positive rate of 17.22%. Additionally, the positive rate of high risk HPV infection was 14.57% and the positive rate of single genotype HPV infection was 13.63%. Totally, 12, 360 HPV viruses were detected. Among them, 10, 879 HPV viruses were classified into high risk genotypes (10, 879 out of 12, 360, 88.02%). The genotypes in women with ages less than 20 were 16/11/6/51/58/52 genotypes; the susceptible HPV genotypes in other women were 16/58/52/53/39/51/81 genotypes.
Conclusions
HPV infections in Shenyang are mainly infections with high risk viruses and single infection. The infection rate and genotype distribution of HPV are different in different age groups. More suitable HPV vaccine prophylaxis can be taken according to the epidemic characteristics of HPV in this area.
10.Protective effect and mechanism of glycyrrhizic acid on the hippocampal neuron injury in epileptic rat models
Qiang WEI ; Jie CHENG ; Yujing HU ; Yanzhu BIAN ; Changzheng DONG
Chinese Journal of Comparative Medicine 2018;28(6):53-58
Objective To explore the protective effect and mechanism of glycyrrhizic acid on hippocampal neuron injury in epileptic rat models. Methods The epileptic rat models were established by lithium and pilocarpine kindling. The successful models were randomly divided into epilepsy group and glycyrrhizic acid group. In addition, the rats without any treatment were used as the normal control group, with 12 rats in each group. Hippocampal neuron injury and apoptosis were detected by Nissl and TUNEL staining. Mitochondrial membrane potential in the hippocampal neurons of rats was detected by JC-1 method. The activity of aspartic acid protein hydrolase 3 (caspase 3) and caspase 9 was detected by colorimetric assay. The expressions of cleaved-caspase 3, 9, B lymphocyte tumor-2 (Bcl-2), Bcl-2 related X protein (Bax), cytochrome C (CytC) and apoptotic protease-activating factor (Apaf-1) protein in the hippocampal tissues of rats were detected by western blot assay. Results Compared with the normal group, the number of neurons was reduced (P<0. 01), the number of TUNEL-positive cells was increased (P< 0. 01), mitochondrial membrane potential was decreased (P< 0. 01), caspase 3 and 9 activity was increased (P< 0. 01), the expressions of Bcl-2 and mitochondrial CytC were down-regulated (P< 0. 01), the expressions of Bax, cytoplasm CytC and Apaf-1 were up-regulated (P< 0. 01) in the epilepsy group. Compared with the epilepsy group, the number of neurons was increased (P < 0. 01), the number of TUNEL-positive cells was reduced (P< 0. 01), mitochondrial membrane potential was increased (P< 0. 01), caspase 3 and 9 activity was decreased (P < 0. 01 ), the expressions of Bcl-2 and mitochondrial CytC were up-regulated (P <0. 01), the expressions of Bax, cytoplasm CytC and Apaf-1 were down-regulated (P < 0. 01) in the glycyrrhizic acid group. Conclusions These results suggest that glycyrrhizic acid can inhibit the hippocampal neuron injury in epileptic rats by blocking the mitochondrial pathway.


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