1.Clinical efficacy of serum beta-amyloid 42 detection and accuracy verification of test kit
Kun JIANG ; Yue TAO ; Minqi CAI ; Dezhu CHEN ; Yiqiang CHEN ; Siyuan LIU ; Bing BAI
Journal of Clinical Medicine in Practice 2025;29(19):1-5,11
Objective To retrospectively analyze the results of serum beta-amyloid 42(Aβ42)tests conducted in the laboratory and explore potential issues associated with this testing.Methods All the Aβ42 test data collected up to the initiation of this study were compiled,and the positive rates for various diagnoses,including Alzheimer's disease(AD),cognitive decline,and dementia were ana-lyzed.Accuracy of the test kit was preliminarily validated using the kit's own reference,mouse brain tissue,and in vitro cultured cell-based reference samples.Results Data from 546 patients were col-lected,with 166 positive cases(positive rate of 30.4%).Among them,3 out of 6 AD patients tested positive;among 149 patients diagnosed with cognitive decline and 59 dementia patients,there were 29 and 24 positive cases respectively,with positive rates of 19.5%and 40.7%.The kit's detection of its own reference was affected by the serum matrix and failed to effectively detect positive mouse brain tissue and cellular reference samples.The kit's detection was generally unaffected by the amyloid precursor protein(APP)of Aβ,but it did not demonstrate good recovery rates for its own reference samples added to various clinical serum specimens.Conclusion The positive rate of serum Aβ42 test results in laboratory is lower than expected,and the accuracy of the test kit may require further improvement.
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.Application of high-frequency ultrasound-guided"cross-shaped"positioning method in open reduction of rib fractures
Tao JI ; Youtao WU ; Biao XIE ; Shuibo ZHU ; Siyuan ZHAN ; Gang YANG ; Yu ZHANG ; Erping XI
Journal of Clinical Surgery 2025;33(7):741-745
Objective To explore the advantages of high-frequency ultrasound-guided"cross-shaped"positioning method in accurate positioning and surgical effect in open reduction and internal fixation of rib fractures.Methods Retrospective analysis of 97 cases of multiple rib fractures treated surgically in our department from october 2019 to october 2024.Patients were divided into three groups based on different localization methods.The control group designed the surgical incision according to the results of chest three-dimensional reconstruction,the ultrasound group determined the incision based on the control group combined with high-frequency ultrasound,and Based on the method used in the ultrasound group,the"cross-localization"technique was combined to determine the incision in the cross-shaped group.The comparison of surgical-related data included fracture positioning accuracy,incision length of per singer rib,postoperative drainage volume,duration of drainage tube retention,postoperative pain score,operative time,intraoperative blood loss,length of hospital stay and incision infection rate.Additionally,commonly used inflammatory markers(white blood cell count,neutrophil percentage,CRP,IL-6,NF-κB)and pain-related biomarkers(PGE2,5-HT)were also included in the comparative analysis.Results In the control group,ultrasound group,and crossover group,aside from wound infections,the respective accuracies of fracture localization were 68.97%vs 87.50%vs 97.22%,the length of single rib incision was(3.50±1.23)cm vs(2.70±0.62)cm vs(2.45±0.58)cm,the volume of drainage on the day of surgery was(170.55±27.85)ml vs(150.69±24.67)ml vs(120.34±18.45)ml,the duration of drainage tube retention was(4.17±1.12)days vs(3.67±0.95)days vs(3.12±0.65)days,the postoperative pain scores were(6.86±2.15)vs(5.54±1.15)vs(4.89±1.53),the operation times were(75.84±15.62)minutes vs(67.74±11.85)minutes vs(57.35±9.36)minutes,the intraoperative blood loss was(85.78±11.78)ml vs(72.65±8.92)ml vs(62.23±9.63)ml,and the length of hospital stay was(8.42±1.47)days vs(7.12±1.14)days vs(6.56±1.32)days.The crossover group showed superior results in all metrics compared to the other two groups,with statistically significant differences(P<0.05).The inflammatory and pain stress indicators for the control,ultrasound,and crossover groups were as follows:white blood cell counts[(16.39±4.15)× 109/Lvs(13.25±2.45)× 109/L vs(12.02±2.77)× 109/L],neutrophil percentages[(80.14±12.21)vs(72.36±10.34)vs(65.73±8.83)],CRP[(31.86±6.87)mg/L vs(27.72±5.65)mg/L vs(24.69±4.11)mg/L],IL-6[(46.35±11.42)pg/L vs(41.42±8.75)pg/L vs(35.53±9.51)pg/L],NF-B[(22.55±4.98)pg/L vs(17.34±3.62)pg/L vs(15.91±3.84)pg/L],PGE2[(240.37±21.65)ng/L vs(209.45±23.24)ng/L vs(180.21±18.72)ng/L],and 5-HT[(290.62±34.37)ng/L vs(270.85±26.98)ng/L vs(210.62±19.64)ng/L].The crossover group demonstrated statistically significant differences compared to both the control and ultrasound groups(P<0.05).There was no statistically significant difference in incision infection among the three groups(P>0.05).Conclusion The ultrasound-guided"cross"positioning method for rib fracture open reduction and internal fixation can more accurately locate the incision,shorten the incision length and hospitalization time,reduce postoperative inflammatory response and patient pain,and facilitate rapid recovery after surgery.
4.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.
5.Application of high-frequency ultrasound-guided"cross-shaped"positioning method in open reduction of rib fractures
Tao JI ; Youtao WU ; Biao XIE ; Shuibo ZHU ; Siyuan ZHAN ; Gang YANG ; Yu ZHANG ; Erping XI
Journal of Clinical Surgery 2025;33(7):741-745
Objective To explore the advantages of high-frequency ultrasound-guided"cross-shaped"positioning method in accurate positioning and surgical effect in open reduction and internal fixation of rib fractures.Methods Retrospective analysis of 97 cases of multiple rib fractures treated surgically in our department from october 2019 to october 2024.Patients were divided into three groups based on different localization methods.The control group designed the surgical incision according to the results of chest three-dimensional reconstruction,the ultrasound group determined the incision based on the control group combined with high-frequency ultrasound,and Based on the method used in the ultrasound group,the"cross-localization"technique was combined to determine the incision in the cross-shaped group.The comparison of surgical-related data included fracture positioning accuracy,incision length of per singer rib,postoperative drainage volume,duration of drainage tube retention,postoperative pain score,operative time,intraoperative blood loss,length of hospital stay and incision infection rate.Additionally,commonly used inflammatory markers(white blood cell count,neutrophil percentage,CRP,IL-6,NF-κB)and pain-related biomarkers(PGE2,5-HT)were also included in the comparative analysis.Results In the control group,ultrasound group,and crossover group,aside from wound infections,the respective accuracies of fracture localization were 68.97%vs 87.50%vs 97.22%,the length of single rib incision was(3.50±1.23)cm vs(2.70±0.62)cm vs(2.45±0.58)cm,the volume of drainage on the day of surgery was(170.55±27.85)ml vs(150.69±24.67)ml vs(120.34±18.45)ml,the duration of drainage tube retention was(4.17±1.12)days vs(3.67±0.95)days vs(3.12±0.65)days,the postoperative pain scores were(6.86±2.15)vs(5.54±1.15)vs(4.89±1.53),the operation times were(75.84±15.62)minutes vs(67.74±11.85)minutes vs(57.35±9.36)minutes,the intraoperative blood loss was(85.78±11.78)ml vs(72.65±8.92)ml vs(62.23±9.63)ml,and the length of hospital stay was(8.42±1.47)days vs(7.12±1.14)days vs(6.56±1.32)days.The crossover group showed superior results in all metrics compared to the other two groups,with statistically significant differences(P<0.05).The inflammatory and pain stress indicators for the control,ultrasound,and crossover groups were as follows:white blood cell counts[(16.39±4.15)× 109/Lvs(13.25±2.45)× 109/L vs(12.02±2.77)× 109/L],neutrophil percentages[(80.14±12.21)vs(72.36±10.34)vs(65.73±8.83)],CRP[(31.86±6.87)mg/L vs(27.72±5.65)mg/L vs(24.69±4.11)mg/L],IL-6[(46.35±11.42)pg/L vs(41.42±8.75)pg/L vs(35.53±9.51)pg/L],NF-B[(22.55±4.98)pg/L vs(17.34±3.62)pg/L vs(15.91±3.84)pg/L],PGE2[(240.37±21.65)ng/L vs(209.45±23.24)ng/L vs(180.21±18.72)ng/L],and 5-HT[(290.62±34.37)ng/L vs(270.85±26.98)ng/L vs(210.62±19.64)ng/L].The crossover group demonstrated statistically significant differences compared to both the control and ultrasound groups(P<0.05).There was no statistically significant difference in incision infection among the three groups(P>0.05).Conclusion The ultrasound-guided"cross"positioning method for rib fracture open reduction and internal fixation can more accurately locate the incision,shorten the incision length and hospitalization time,reduce postoperative inflammatory response and patient pain,and facilitate rapid recovery after surgery.
6.Discovery of A New Prognostic Molecular Marker NKX2-3 for Acute Myeloid Leukemia
Wandi WANG ; Tao CHANG ; Siyuan JIANG ; Qi HOU ; Zhenyi JIN ; Xiuli WU
Journal of Sun Yat-sen University(Medical Sciences) 2024;45(1):63-68
ObjectiveTo analyze the expression of molecular marker affecting the prognosis of acute myeloid leukemia (AML) patients from bioinformatics database, thus providing an experimental basis for further exploration of a novel molecular marker for the prognosis of AML. MethodsThe prognostic data of 179 AML patients from The Cancer Genome Atlas (TCGA) database were examined for differential gene analysis and survival analysis. The bone marrow samples of 74 healthy individuals (HI) and 542 de novo AML patients in the dataset GSE13159 downloaded from the Gene Expression Omnibus (GEO) database were analyzed to detect the difference in the expression levels of differential target genes. Peripheral blood and bone marrow samples were collected from 18 de novo AML patients and 20 age- and gender-matched healthy controls, and real-time fluorescent quantitative PCR was used to validate the expression levels of the differential genes in the AML patients. ResultsBioinformatics data analysis showed that the optimal cut-off value of Homo sapiens NK2 homeobox 3 (NKX2-3) calculated by R language was 0.051. Survival analysis revealed a statistically poorer overall survival in de novo AML patients with high NKX2-3 expression than in those with low NKX2-3 expression (P = 0.0036). NKX2-3 was highly expressed in patients with de novo AML than in HI and the difference was statistically significant (P < 0.001). Real-time fluorescence quantitative PCR verified the expression levels of the NKX2-3 gene in AML patients and confirmed that compared with those in HI, in the de novo AML patients, NKX2-3-1 and NKX2-3-2 were highly expressed and were significantly correlated (P = 0.000, P = 0.000). ConclusionNKX2-3 is highly expressed in de novo AML patients, and the AML patients with high NKX2-3 expression have poor overal survival. NKX2-3 may be closely related to the clinical outcome and prognosis of AML.
7.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.
8.Advances in preoperative predictive indicators for microvascular invasion in hepatocellular carcinoma
Siqiao SHAN ; Siyuan WANG ; Dongliang YANG ; Nan JIANG ; Mingyu LIN ; Tao ZHANG ; Xueli YUAN ; Shuo JIN ; Jianping ZENG
Chinese Journal of Hepatobiliary Surgery 2024;30(9):705-709
Hepatocellular carcinoma (HCC) is characterized by high postoperative recurrence and mortality rates. In recent years, researchers have identified a significant correlation between microvascular invasion (MVI) and early postoperative recurrence and metastasis of HCC, making it a focal point of HCC research. Accurate preoperative prediction of MVI occurrence and the implementation of relevant interventions (such as expanded resection) could provide substantial benefits to patients. This study analyzes global research over the past decade on MVI predictive indicators based on tumor biological characteristics, genetic measurements, imaging examinations, and tumor markers. The aim is to use these predictive indicators to objectively forecast the occurrence of MVI, thereby aiding in preoperative individual assessments and enhancing treatment plans.
9.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.
10.Discussion on the Treatment of Malignant Tumors by the Method of Counteracting Toxin with Toxin
Siyuan XU ; Liu LI ; Yueyang LAI ; Lihuiping TAO ; Junyi WANG ; Xiaoman WEI ; Dongdong SUN ; Haibo CHENG
Journal of Traditional Chinese Medicine 2023;64(18):1847-1852
Cancer toxin is the key pathogenesis of malignant tumors. The basic principle of cancer treatment is “dispelling pathogen and resolving toxins, reinforcing healthy qi and reinforcing the foundation”. As one of the “eight methods of anticancer and detoxification”, the counteracting toxin with toxin therapy is a commonly used clinical treatment of malignant tumors. This paper discussed the method of counteracting toxin with toxin and its application in the prevention and treatment of malignant tumors from the aspects of history tracing, academic connotation, application principles and clinical application. Toxic Chinese medicinals with anticancer function are required to eliminate cancer toxins based on the principles of excessive cancer toxicity and plentiful healthy qi, as well as in accordance with the various stages and classifications of tumors, thereby improving the theoretical connotation of the method of counteracting toxin with toxin, and promoting the popularization and application of the pathogenesis theory of cancer toxin in the prevention and treatment of malignant tumors.

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