1.Transcriptomics of the Lieber-DeCarli mouse model of alcoholic liver injury
Tianyin RUAN ; Siyuan WANG ; Xutao LI ; Hao ZHANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Acta Laboratorium Animalis Scientia Sinica 2025;33(2):204-215
Objective To investigate the characteristics of liver injury in the Lieber-DeCarli alcoholic liver disease(ALD)mouse model and to analyze its transcriptomic profile.Methods Eighteen male C57BL/6J mice were randomly divided into an alcohol-fed group(n = 10)and a control group(n = 8).The alcohol-fed group received a Lieber-DeCarli ethanol diet,starting with an adaptive one-week phase using incremental concentrations of ethanol(10~57.3 mL/L),followed by 2 weeks of a 57.3 mL/L concentration of 95%ethanol,for a total of 3 weeks.The control group was provided with an isocaloric control diet for 3 weeks.At the end of the study,mice were sacrificed,and serum and liver tissue samples were collected.Serum liver function markers(ALT,AST),hepatic lipids(TC,TG),reduced glutathione(GSH),total superoxide dismutase(T-SOD),and malondialdehyde(MDA)were measured using biochemical assays.The levels of inflammatory cytokines(IL-6,IL-10,TNF-α,TGF-β1)in liver tissue were assessed by ELISA.Histopathological changes in liver tissue were examined using hematoxylin-eosin(HE)and Oil Red O staining.Immunohistochemical staining using the F4/80 antibody was employed to assess changes in macrophage expression.RNA-seq analysis was conducted to identify differentially expressed genes between the two groups of liver tissues,followed by GO and KEGG pathway enrichment analysis.qRT-PCR was used to validate the expression of these differentially expressed genes.Results Compared with the control group,the alcohol-fed mice exhibited a significant decrease in body weight(P<0.01).Serum ALT and AST levels were significantly elevated(P<0.01),while liver tissue levels of TC,TG,and MDA were significantly increased(P<0.05).Conversely,GSH and T-SOD levels were significantly reduced(P<0.05).The levels of inflammatory factors IL-6,TNF-α,and TGF-β1 were increased,which was consistent with the qRT-PCR validation results(P<0.05).Histological examination revealed disrupted hepatic lobular structure,with macrovesicular steatosis,microvesicular steatosis,and ballooning degeneration.Additionally,fat droplets in liver tissue were significantly increased,and macrophage expression was upregulated.Differential gene expression analysis,using a threshold of|log2 FC|>1 and q<0.05,identified 2063 differentially expressed genes,of which 1236 were upregulated and 827 downregulated.Enriched pathways included xenobiotic metabolism via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,steroid hormone biosynthesis,glutathione metabolism,and retinol metabolism.(P<0.05).qRT-PCR validation confirmed the significant upregulation(e.g.,Mmp12,Gstm3,Cyp2a22)and downregulation(e.g.,Serpina1e,Acmsd,Mup3d)of 10 genes from each category,consistent with the transcriptome sequencing results.Conclusions The primary pathological mechanisms underlying alcoholic liver injury involve pathways related to xenobiotic metabolism and act via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,glutathione metabolism,and retinol metabolism.
2.Transcriptomics of the Lieber-DeCarli mouse model of alcoholic liver injury
Tianyin RUAN ; Siyuan WANG ; Xutao LI ; Hao ZHANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Acta Laboratorium Animalis Scientia Sinica 2025;33(2):204-215
Objective To investigate the characteristics of liver injury in the Lieber-DeCarli alcoholic liver disease(ALD)mouse model and to analyze its transcriptomic profile.Methods Eighteen male C57BL/6J mice were randomly divided into an alcohol-fed group(n = 10)and a control group(n = 8).The alcohol-fed group received a Lieber-DeCarli ethanol diet,starting with an adaptive one-week phase using incremental concentrations of ethanol(10~57.3 mL/L),followed by 2 weeks of a 57.3 mL/L concentration of 95%ethanol,for a total of 3 weeks.The control group was provided with an isocaloric control diet for 3 weeks.At the end of the study,mice were sacrificed,and serum and liver tissue samples were collected.Serum liver function markers(ALT,AST),hepatic lipids(TC,TG),reduced glutathione(GSH),total superoxide dismutase(T-SOD),and malondialdehyde(MDA)were measured using biochemical assays.The levels of inflammatory cytokines(IL-6,IL-10,TNF-α,TGF-β1)in liver tissue were assessed by ELISA.Histopathological changes in liver tissue were examined using hematoxylin-eosin(HE)and Oil Red O staining.Immunohistochemical staining using the F4/80 antibody was employed to assess changes in macrophage expression.RNA-seq analysis was conducted to identify differentially expressed genes between the two groups of liver tissues,followed by GO and KEGG pathway enrichment analysis.qRT-PCR was used to validate the expression of these differentially expressed genes.Results Compared with the control group,the alcohol-fed mice exhibited a significant decrease in body weight(P<0.01).Serum ALT and AST levels were significantly elevated(P<0.01),while liver tissue levels of TC,TG,and MDA were significantly increased(P<0.05).Conversely,GSH and T-SOD levels were significantly reduced(P<0.05).The levels of inflammatory factors IL-6,TNF-α,and TGF-β1 were increased,which was consistent with the qRT-PCR validation results(P<0.05).Histological examination revealed disrupted hepatic lobular structure,with macrovesicular steatosis,microvesicular steatosis,and ballooning degeneration.Additionally,fat droplets in liver tissue were significantly increased,and macrophage expression was upregulated.Differential gene expression analysis,using a threshold of|log2 FC|>1 and q<0.05,identified 2063 differentially expressed genes,of which 1236 were upregulated and 827 downregulated.Enriched pathways included xenobiotic metabolism via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,steroid hormone biosynthesis,glutathione metabolism,and retinol metabolism.(P<0.05).qRT-PCR validation confirmed the significant upregulation(e.g.,Mmp12,Gstm3,Cyp2a22)and downregulation(e.g.,Serpina1e,Acmsd,Mup3d)of 10 genes from each category,consistent with the transcriptome sequencing results.Conclusions The primary pathological mechanisms underlying alcoholic liver injury involve pathways related to xenobiotic metabolism and act via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,glutathione metabolism,and retinol metabolism.
3.Association of sleep status with ambulatory blood pressure monitoring indicators in community-dwelling hypertensive patients
Hongmei ZHANG ; Lanping CAI ; Yajuan WANG ; Ling CHEN ; Yanyan ZHOU ; Haiyan LENG ; Tiemei RUAN ; Xiaoying TANG ; Yu FENG ; Xue BAI ; Puyang ZHENG
Chinese Journal of General Practitioners 2024;23(12):1262-1269
Objective:To analyze the association between sleep status and ambulatory blood pressure monitoring indicators in community-dwelling hypertensive patients.Methods:It was a cross sentional study. Hypertensive patients who underwent 24-hour ambulatory blood pressure monitoring from May 2021 to April 2023 in Shanghai Xinzhuang Town were enrolled. The demographic information and sleep status of patients were obtained from the questionnaire. A TM-2430 blood pressure monitor was used to measure 24-hour ambulatory blood pressure, and the relevant indicators, including blood pressure level and blood pressure coefficient of variation were documented. The association between sleep status and blood pressure indicators was analyzed with multivariate linear regression model.Results:A total 1 135 patients aged (65.07±12.61) years were enrolled, and 473 (41.67%) of whom were males. The sleep time was<7 hours in 76 cases, 7- 8 hours in 219 cases and >8 hours in 840 cases; the bedtime was earlier than 22∶00 in 415 cases, between 22∶00 and 23∶00 in 474 cases and later than 23∶00 in 246 cases; the wake-up time was before 6∶00 in 230 cases, between 6∶00 and 7∶00 in 521 cases and after 7∶00 in 384 cases. Multivariate linear regression analysis showed that after controlling for gender and age, the sleep time was negatively associated with diurnal, noctumal and 24-hour diastolic blood pressure levels (all P<0.05), and positively associated with diurnal and noctumal systolic blood pressure coefficient of variation, noctumal diastolic blood pressure coefficient of variation, and 24-hour systolic blood pressure coefficient of variation (all P<0.05).The bedtime was positively associated with diurnal, noctumal and 24-hour diastolic blood pressure (all P<0.05), diastolic blood pressure (all P<0.05); and negatively associated with diurnal systolic blood pressure coefficient of variation, diurnal diastolic blood pressure coefficient of variation, noctumal systolic blood pressure coefficient of variation, 24-hour systolic blood pressure coefficient of variation, and 24-hour diastolic blood pressure coefficient of variation (all P<0.05). The wake-up time was positively associated with diurnal systolic blood pressure, diurnal, noctumal and 24-hour diastolic blood pressure (all P<0.05), and positively associated with diurmal systolic blood pressure and diastolic blood pressure (both P<0.05). Conclusion:Sleep status is closely associated with ambulatory blood pressure monitoring indicators in community-dwelling hypertensive patients.
4.Transcriptomics-based analysis of the effect of compound Dancao granules on hepatocyte apoptosis in mice with non-alcoholic steatohepatitis
Xutao LI ; Siyuan WANG ; Tianyin RUAN ; Hao ZHANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Acta Laboratorium Animalis Scientia Sinica 2024;32(12):1543-1555
Objective Transcriptome sequencing technology(RNA-seq)was used to analyze the mechanism of compound Dancao granules as an intervention for high-fat feed combined with carbon tetrachloride(CCl4)-induced non-alcoholic steatohepatitis.Methods 45 male C57BL/6J mice were split into two groups at random:normal control group,model control group,obeccholic acid group 10 mg/(kg·d),and compound Dancao granules low-and high-dose groups 3.74 g/(kg·d)and 7.48 g/(kg·d),with 9 mice in each group.Normal diet was made available to the control group,and the mice in the model group were given a high-fat diet combined with the subcutaneous injection of CC14,with 100%CC14 solution(4 mL/kg)in the first application,and 40%CC14-olive oil solution(2 mL/kg)in the second application,twice a week for a total of 6 weeks.Each drug group was administered the respective drug from week 3 for a total of 4 weeks.12 h after the last administration,the serum and liver tissues of mice in each group were collected,and a biochemical kit was used to detect serum liver function.Hematoxylin-eosin(HE),sirius scarlet,and oil red O staining were used to examine histopathological changes to the liver.The levels of IL-6,IL-10,TNF-α and TGF-β in mice liver were detected via ELISA,and the expression of α-SMA was observed by immunohistochemistry.Differential gene expression was analyzed by RNA-seq and functional enrichment analysis.To verify the differential expression of mRNA,quantitative reverse transcription PCR(qRT-PCR)was used.TDT-mediated dUTP nick-end labeling(TUNEL)staining was employed to identify apoptosis.Results The model control groups had significantly higher levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),and triglycerides(TG)than normal control group(P<0.01).Additionally,there was obvious inflammatory cell infiltration in the liver tissue,collagen deposition in the sink and interlobule areas,and a significant increase in lipid droplet area(P<0.01).The levels of IL-6 and TNF-α in liver tissue were significantly increased(P<0.01),the levels of IL-10 and TGF-β were decreased(P<0.01),and the expression of α-SMA was significantly increased(P<0.01).The levels of TC,TG,ALT,and AST were significantly lower in groups that received compound Dancao granules and obeccholic acid than the model control group(P<0.01),and inflammatory cell infiltration,collagen deposition,and fat accumulation in the sink and interlobule areas were improved(P<0.01).The levels of IL-6 and TNF-α in liver tissue were significantly decreased(P<0.01),the levels of IL-10 and TGF-β were increased(P<0.05,P<0.01),and the expression of α-SMA was significantly decreased(P<0.01).RNA-seq sequencing result showed that 2819 genes in the normal control group were differentially expressed compared with the model control group,with 543 up-regulated and 2276 down-regulated genes.In a comparison of the model control group and compound Dancao granules group,240 genes were differentially expressed,including 206 up-regulated genes and 34 down-regulated genes.There were 221 genes with overlapping expression in the 2 groups and functional enrichment highlighted cell cycle(Cdt1,Plk1,Bub1b,Ttk,Knl1,Esco2,Cdc6,Ndc80,Cdc25b,Sgo1,Ccnb2,Espl1,Ccne1,Mcm4,Mcm5,Fbxo5,Bub1,Mcm2),apoptosis(Caspase3,Bax,P53,Apaf1,Bak,Caspase8),the P53 signaling pathway(P53,Ccnb2,Apaf1,Bak,Bax,Gtse1,Caspase3,Ccne1),arachidonic acid metabolism(Hpgds,Cyp2c54,Cyp2b10,Tbxas1,Cyp2c50),galactose metabolism(Hk3,Gla,Hk2,Akr1b7)and other signaling pathway genes.RNA-seq sequencing analysis showed that compound Danicao granules mainly regulated the apoptosis signaling pathway,and qRT-PCR confirmed that the mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in the liver tissue of the model control group was increased compared with that of the normal control group(P<0.01).Compared with the model control group,the compound Dancao granules group showed decreased mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in liver tissue(P<0.01).TUNEL staining showed that the number of cells showing nuclear shrinkage and apoptotic bodies decreased in the compound Dancao granule administration group.Conclusions Compound Dancao granules had a significant protective effect against non-alcoholic steatohepatitis induced by high-fat feed combined with CCl4,and its mechanism might be connected to the control of genes linked to apoptosis.
5.Transcriptomics-based analysis of the effect of compound Dancao granules on hepatocyte apoptosis in mice with non-alcoholic steatohepatitis
Xutao LI ; Siyuan WANG ; Tianyin RUAN ; Hao ZHANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Acta Laboratorium Animalis Scientia Sinica 2024;32(12):1543-1555
Objective Transcriptome sequencing technology(RNA-seq)was used to analyze the mechanism of compound Dancao granules as an intervention for high-fat feed combined with carbon tetrachloride(CCl4)-induced non-alcoholic steatohepatitis.Methods 45 male C57BL/6J mice were split into two groups at random:normal control group,model control group,obeccholic acid group 10 mg/(kg·d),and compound Dancao granules low-and high-dose groups 3.74 g/(kg·d)and 7.48 g/(kg·d),with 9 mice in each group.Normal diet was made available to the control group,and the mice in the model group were given a high-fat diet combined with the subcutaneous injection of CC14,with 100%CC14 solution(4 mL/kg)in the first application,and 40%CC14-olive oil solution(2 mL/kg)in the second application,twice a week for a total of 6 weeks.Each drug group was administered the respective drug from week 3 for a total of 4 weeks.12 h after the last administration,the serum and liver tissues of mice in each group were collected,and a biochemical kit was used to detect serum liver function.Hematoxylin-eosin(HE),sirius scarlet,and oil red O staining were used to examine histopathological changes to the liver.The levels of IL-6,IL-10,TNF-α and TGF-β in mice liver were detected via ELISA,and the expression of α-SMA was observed by immunohistochemistry.Differential gene expression was analyzed by RNA-seq and functional enrichment analysis.To verify the differential expression of mRNA,quantitative reverse transcription PCR(qRT-PCR)was used.TDT-mediated dUTP nick-end labeling(TUNEL)staining was employed to identify apoptosis.Results The model control groups had significantly higher levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),and triglycerides(TG)than normal control group(P<0.01).Additionally,there was obvious inflammatory cell infiltration in the liver tissue,collagen deposition in the sink and interlobule areas,and a significant increase in lipid droplet area(P<0.01).The levels of IL-6 and TNF-α in liver tissue were significantly increased(P<0.01),the levels of IL-10 and TGF-β were decreased(P<0.01),and the expression of α-SMA was significantly increased(P<0.01).The levels of TC,TG,ALT,and AST were significantly lower in groups that received compound Dancao granules and obeccholic acid than the model control group(P<0.01),and inflammatory cell infiltration,collagen deposition,and fat accumulation in the sink and interlobule areas were improved(P<0.01).The levels of IL-6 and TNF-α in liver tissue were significantly decreased(P<0.01),the levels of IL-10 and TGF-β were increased(P<0.05,P<0.01),and the expression of α-SMA was significantly decreased(P<0.01).RNA-seq sequencing result showed that 2819 genes in the normal control group were differentially expressed compared with the model control group,with 543 up-regulated and 2276 down-regulated genes.In a comparison of the model control group and compound Dancao granules group,240 genes were differentially expressed,including 206 up-regulated genes and 34 down-regulated genes.There were 221 genes with overlapping expression in the 2 groups and functional enrichment highlighted cell cycle(Cdt1,Plk1,Bub1b,Ttk,Knl1,Esco2,Cdc6,Ndc80,Cdc25b,Sgo1,Ccnb2,Espl1,Ccne1,Mcm4,Mcm5,Fbxo5,Bub1,Mcm2),apoptosis(Caspase3,Bax,P53,Apaf1,Bak,Caspase8),the P53 signaling pathway(P53,Ccnb2,Apaf1,Bak,Bax,Gtse1,Caspase3,Ccne1),arachidonic acid metabolism(Hpgds,Cyp2c54,Cyp2b10,Tbxas1,Cyp2c50),galactose metabolism(Hk3,Gla,Hk2,Akr1b7)and other signaling pathway genes.RNA-seq sequencing analysis showed that compound Danicao granules mainly regulated the apoptosis signaling pathway,and qRT-PCR confirmed that the mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in the liver tissue of the model control group was increased compared with that of the normal control group(P<0.01).Compared with the model control group,the compound Dancao granules group showed decreased mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in liver tissue(P<0.01).TUNEL staining showed that the number of cells showing nuclear shrinkage and apoptotic bodies decreased in the compound Dancao granule administration group.Conclusions Compound Dancao granules had a significant protective effect against non-alcoholic steatohepatitis induced by high-fat feed combined with CCl4,and its mechanism might be connected to the control of genes linked to apoptosis.
6. Protection management and procedures of nuclear medicine imaging during novel coronavirus (2019-nCov) infection epidemic period
Xiaoli LAN ; Xun SUN ; Chunxia QIN ; Weiwei RUAN ; Jia HU ; Jing LIN ; Fan HU ; Ting WANG ; Xiaotian XIA ; Yongxue ZHANG ; Rui AN ; Zairong GAO ; Yanyan WU ; Lijuan XIONG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2020;40(2):105-107
At the end of December 2019, acute respiratory infectious diseases caused by a new type of coronavirus were prevalent in Wuhan and other cities of China. Different from radiology examinations, the protocols of nuclear medical imaging examinations are complicated, in which more workplaces and staff are needed, resulting more complex management of patients and higher protection requirements. Combined with the characteristics of SPECT and PET imaging procedures, this paper puts forward some suggestions on the protective process of imaging examinations for patients with confirmed or suspected novel coronavirus infection. The main purpose is to protect medical staff from virus infection, effectively reduce the risk of virus transmission during the examination process, and ensure the medical quality and safety.
7. Protection management and procedures of nuclear medicine imaging during novel coronavirus (2019-nCov) infection epidemic period
Xiaoli LAN ; Xun SUN ; Chunxia QIN ; Weiwei RUAN ; Jia HU ; Jing LIN ; Fan HU ; Ting WANG ; Xiaotian XIA ; Yongxue ZHANG ; Rui AN ; Zairong GAO ; Yanyan WU ; Lijuan XIONG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2020;40(0):E001-E001
At the end of December 2019, acute respiratory infectious diseases caused by a new type of coronavirus were prevalent in Wuhan and other cities of China. Different from radiology examinations, the protocols of nuclear medical imaging examinations are complicated, in which more workplaces and staff are needed, resulting more complex management of patients and higher protection requirements. Combined with the characteristics of SPECT and PET imaging procedures, this paper puts forward some suggestions on the protective process of imaging examinations for patients with confirmed or suspected noval coronavirus infec- tion. The main purpose is to protect medical staff from virus infection, effectively reduce the risk of virus transmission during the examination process, and ensure the medical quality and safety.
8.Experience and needs of symptom management care for parents of children with cancer during chemotherapy: a qualitative study
Yanyan LIU ; Nanping SHEN ; Jing SUN ; Haishan RUAN ; Yiming LIU
Chinese Journal of Modern Nursing 2020;26(35):4914-4918
Objective:To explore the experience and needs of symptom management care of parents of cancer children during chemotherapy so as to provide a reference for improving the level of symptom management.Methods:Between November 2018 and April 2019, phenomenological research method was used to conduct a semi-structured interview with 14 parents of cancer children in the stage of chemotherapy from two hospitals. Colaizzi seven step analysis was used to the data analysis.Results:The care experience of symptoms and adverse reactions of parents of children with cancer during chemotherapy was refined into two themes, namely the process of learning and growth, and the transformation of psychological adaptation. The main needs of care were information needs, communication needs and psychological support needs.Conclusions:Parents of children with cancer are learning and growing up in the process of treatment and symptom care, and there are many needs. Medical and nursing staff should work closely with the children and their parents to meet the parents' needs for knowledge, communication and psychological support for symptom care, improve the care experience of parents and alleviate the pain of children.
9.Association study of gamma-aminobutyric acid type A receptor genes cluster with autism in Chinese Han population
Zhiliu WU ; Jun XIONG ; Linyan WANG ; Tianlan LU ; Yanyan RUAN ; Weihua YUE ; Dai ZHANG ; Lifang WANG
Chinese Mental Health Journal 2018;32(1):83-88
Objective:To investigate the genetic association of single nucleotide polymorphisms (SNPs) in gamma-aminobutyric acid type A (GABAA) receptor genes cluster on chromosome 15q12 with autism in Chinese Han population.Methods:Totally 502 autism trios of Chinese Han ethnicity (including 502 autism individuals and 1004 healthy biological parents) were selected.All children met the autism diagnosis of Diagnostic and Statistical Manual of Mental Disorders,Fourth edition (DSM-Ⅳ).Genotyping for 15 selected tag SNPs in three GABAA receptor genes (GABRB3,GABRA5,and GABRG3) was performed using Agena Bioscience MassARRAY platform.The family-based association test for 15 tag SNPs was performed to compare the transmitted frequency of al leles of heterozygous genotypes from parents to offspring in autism trios.Results:The C allele of rs7180500 in GABRG3 and the A allele of rs4906902 in GABRB3 exhibited the preferential transmission from parents to affected offspring (Z =3.573,P <0.001;Z =3.141,P =0.002),and the association was significant after Bonferroni correction.Conclusion:It suggests that GABRG3 and GABRB3 which located in chromosome 15q12 might be susceptibility genes in Chinese Han population.
10."Research on the new model and scheme under the background of ""dual-track to one"" training mode on the acupuncture standardization training of Chinese medicine master's degree"
Can DONG ; Desong KONG ; Yanyan XU ; Zhilan HUANG ; Jinjin PAN ; Xiaosu ZHANG ; Zhizhong RUAN
International Journal of Traditional Chinese Medicine 2017;39(3):254-257
Under the background of dual-track training mode, the current clinical program of professional master's graduate education lacks complete and unified content and requirements. We set out a unified teaching scheme of subspecialty, in the main subject acupuncture. We identified the teaching management and innovated the teaching and examination methods in the scheme. Our research gradually formed a multidisciplinary talents training system, as the foundation of developing the standardization training. This method could help to cultivate qualified graduates as the high quality talents for the national medicine development.

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