1.In Vitro and In Vivo Chemical Composition Analysis of Reference Sample of Jinshui Liujunjian Based on UPLC-Q-TOF-MS/MS
Xinyue YANG ; Huiyu LI ; Yaqi LOU ; Xingxing WANG ; Guifang YU ; Chenfeng ZHANG ; Zhenzhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):166-173
ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was used to detect the reference sample solution, plasma, and tissue samples of Jinshui Liujunjian under positive and negative ion modes, respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times, secondary mass spectral fragments, reference standards, and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian, including 47 flavonoids, 5 amino acids, 13 iridoids, 16 triterpenoid saponins, etc., of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components; 50 prototype components were identified in different tissues, among which 13, 10, 7, 21, 11, 6, 14, and 40 prototype components were identified in the heart, liver, spleen, lung, kidney, brain, large intestine, and stomach, respectively. Among them, 7 compounds such as ferulic acid, glycyrrhizic acid, and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian, primarily composed of flavonoids and triterpenoid saponins, along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.
2.In Vitro and In Vivo Chemical Composition Analysis of Reference Sample of Jinshui Liujunjian Based on UPLC-Q-TOF-MS/MS
Xinyue YANG ; Huiyu LI ; Yaqi LOU ; Xingxing WANG ; Guifang YU ; Chenfeng ZHANG ; Zhenzhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):166-173
ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was used to detect the reference sample solution, plasma, and tissue samples of Jinshui Liujunjian under positive and negative ion modes, respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times, secondary mass spectral fragments, reference standards, and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian, including 47 flavonoids, 5 amino acids, 13 iridoids, 16 triterpenoid saponins, etc., of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components; 50 prototype components were identified in different tissues, among which 13, 10, 7, 21, 11, 6, 14, and 40 prototype components were identified in the heart, liver, spleen, lung, kidney, brain, large intestine, and stomach, respectively. Among them, 7 compounds such as ferulic acid, glycyrrhizic acid, and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian, primarily composed of flavonoids and triterpenoid saponins, along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.
3.Value of preoperative ultrasound combined with NLR,Ctn and CEA levels in predicting cervical lymph node metastasis of papillary thyroid carcinoma
Shanshan YAN ; Huiyu ZHANG ; Jun GU ; Kan ZHANG ; Yu ZHAO
The Journal of Practical Medicine 2025;41(18):2859-2864
Objective To investigate the predictive value of preoperative ultrasound in combination with neutrophil-to-lymphocyte ratio(NLR),calcitonin(Ctn),and carcinoembryonic antigen(CEA)for cervical lymph node metastasis in patients with papillary thyroid carcinoma.Methods A total of 103 patients diagnosed with papillary thyroid carcinoma(PTC)who were admitted to the hospital between October 2021 and October 2024 were selected as the case group.Among them,34 patients with cervical lymph node metastasis confirmed by surgical and pathological examination were assigned to the metastatic group,and 69 patients without cervical lymph node metas-tasis were assigned to the non-metastatic group.Additionally,103 patients with benign thyroid nodules admitted during the same period were enrolled as the control group.Clinical data,ultrasonographic features,and serum levels of NLR,Ctn,and CEA were compared between the metastatic and non-metastatic groups.The predictive value of ultrasonographic features and the combined detection of NLR,Ctn,and CEA levels for cervical lymph node metastasis in PTC was evaluated using receiver operating characteristic(ROC)curve analysis.Results Compared to the control group,the case group exhibited a higher proportion of patients with microcalcification and grade 3 blood flow,as well as elevated levels of NLR,Ctn,and CEA(P<0.05).Similarly,compared to the non-metastatic group,the metastatic group showed a higher proportion of patients with microcalcification and grade 3 blood flow,along with increased levels of NLR,Ctn,and CEA(P<0.05).The metastatic group tested positive,whereas the non-metastatic group tested negative.The area under the curve(AUC)for ultrasound features(micro-calcification,blood flow classification)combined with NLR,Ctn,and CEA levels in diagnosing cervical lymph node metastasis in papillary thyroid carcinoma was higher than that of individual indicators(P<0.05).Conclusions Preoperative ultrasound combined with the assessment of NLR,Ctn,and CEA levels demonstrates significant predictive value for cervical lymph node metastasis in papillary thyroid carcinoma.
4.Effect of C1q tumor necrosis factor-related protein 3 on reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells by Sendai virus vector overexpressing Gata4,Mef2c,and Tbx5
Yanbin SONG ; Yunqing ZHANG ; Huiyu LIU ; Junming CHEN
Chinese Journal of Comparative Medicine 2025;35(8):14-28
Objective To investigate the efficiency and mechanism of C1q tumor necrosis factor-related protein 3(CTRP3)on reprogramming of cardiac fibroblasts(CFs)into induced cardiomyocyte-like cells(iCMs)by Sendai virus(SeV)vector overexpressing Gata4,Mef2c and Tbx5(SeVGMT).Methods CFs were divided into Control,NC-Lv,CTRP3-Lv,NC-sh,and CTRP3-sh groups.NC-Lv,CTRP3-Lv,NC-sh,and CTRP3-sh were transfected into CFs using Lipofectamine 3000 reagent for 48 hours.Lipofectamine 3000 reagent was then mixed with SeVGMT and incubated at room temperature for 48 hours,the culture medium was then replaced,and cells were cultured for 21 days.Cell morphology was observed under a microscope at 0,3,7,14,and 21 days.Expression levels of the myocardial-specific proteins α-myosin heavy chain(α-MHC),α-actin,cardiac troponin T(cTnT),connexin 43(Cx43),cardiac muscle α-actin(Actc1),and myosin heavy chain 6(Myh6)were detected at different time points by immunofluorescence,quantitative reverse transcription-polymerase chain reaction,and Western blot,and the proportions of beating cells at different time points were calculated.Results The relative fluorescence intensity and mRNA and protein levels of α-MHC,α-actin,cTnT,Cx43,Actc1,and Myh6 in CFs in each group increased with increasing culture time(P<0.05),with significantly higher expression levels of myocardial-specific proteins at 14 days of culture than at 7 days(P<0.05).The relative fluorescence intensities and mRNA and protein levels of α-MHC,α-actin,cTnT,Cx43,Actc1,and Myh6 in CFs at 3,7,14,and 21 days of culture were significantly increased in the CTRP3-Lv group compared with the NC-Lv group(P<0.05),but were significantly decreased in CFs in the CTRP3-sh group compared with the NC-sh group(P<0.05).Beating cells appeared in CFs in each group at 7 days of culture.The proportion of beating cells in each group increased with increasing culture time(P<0.05),and the proportion was significantly higher at 14 days than at 7 days(P<0.05).The proportion of beating cells among CFs was increased in the CTRP3-Lv group at 7,14,and 21 days of culture compared with the NC-Lv group(P<0.05),while the proportion of beating cells in the CTRP3-sh group was decreased compared with the NC-sh group(P<0.05).Conclusions CTRP3 can enhance SeVGMT reprogramming of CFs into iCMs.
5.Fasudil alleviates beta-amyloid 1-42-induced apoptosis of SH-SY5Y cells
Minfang GUO ; Huiyu ZHANG ; Peijun ZHANG ; Qin SU ; Siwei JIA ; Jiezhong YU
Chinese Journal of Tissue Engineering Research 2025;29(23):4939-4946
BACKGROUND:Fasudil has a regulatory effect on mitochondrial dynamics in the brain of Alzheimer's disease mice and can inhibit neuroinflammation,but whether it can reduce the toxicity of β-amyloid protein by regulating mitophagy-NLRP3 inflammasome pathway remains unclear.OBJECTIVE:To investigate the regulatory effect of fasudil on β-amyloid 1-42-induced apoptosis and mitophagy and NLRP3 inflammasome in human derived neuroblastoma cell line SH-SY5Y cells.METHODS:SH-SY5Y cells were inoculated into the pore plate.After adhesion,cells were divided into three groups for intervention:No drug was added to the control group;20 μmol/L β-amyloid 1-42 was added to the model group,and 20 μmol/L β-amyloid 1-42 and 15 mg/L fasudil were added to the fasudil group at the same time.After 24 hours of intervention,the cell activity was detected by MTT assay and apoptosis was detected by TUNEL staining.The expression of apoptosis-related proteins was detected by qRT-PCR and western blot assay.The expression of mitochondrial autophagy related proteins was detected by immunofluorescence staining and western blot assay.The expression of NLRP3 inflammasome related proteins was detected by immunofluorescence staining and western blot assay.RESULTS AND CONCLUSION:(1)Compared with control group,the cell activity of the model group was decreased and the apoptosis rate was increased(P<0.05).Compared with model group,cell activity in the fasudil group was increased and apoptosis rate was decreased(P<0.05).(2)The results of qRT-PCR and western blot assay showed that compared with the control group,the expression of Bax mRNA and protein was increased in the model group(P<0.05),while the expression of Bcl-2 mRNA and protein was decreased(P<0.05).Compared with the model group,the expression of Bax mRNA and protein was decreased(P<0.05),and the expression of Bcl-2 mRNA and protein was increased(P<0.05)in the fasudil group.(3)The results of immunofluorescence staining and western blot assay showed that compared with the control group,the expressions of PINK1,Parkinson's disease protein and LC3 protein were decreased(P<0.05),while the expression of p62 protein was increased(P<0.05)in the model group.Compared with model group,the expression levels of PINK1,Parkinson's disease protein,and LC3 protein were increased(P<0.05),while the expression of p62 protein was decreased(P<0.05)in the fasudil group.(4)The results of immunofluorescence staining and western blot assay showed that compared with the control group,the expression levels of NLRP3,ASC,Caspase-1,and interleukin1β protein were increased in the model group(P<0.05).Compared with the model group,the expression levels of NLRP3,ASC,Caspase-1,and interleukin1β were decreased in the fasudil group(P<0.05).(5)The results show that fasudil can reduce the apoptosis of SH-SY5Y cells induced by β-amyloid 1-42,and its mechanism may be related to the activation of mitophagy and the inhibition of NLRP3 inflammasome activation.
6.Intraoperative ultrasound for microsurgical resection of primary supratentorial glioblastoma
Fangxian LI ; Huiyu MA ; Wei ZHANG ; Wen HE
Chinese Journal of Medical Imaging Technology 2025;41(4):583-586
Objective To observe the value of intraoperative ultrasound(IOUS)for microsurgical resection of primary supratentorial glioblastoma.Methods Totally 130 cases of central nervous system WHO grade 4 primary supratentorial glioblastoma confirmed by postoperative pathology were retrospectively enrolled and divided into IOUS group and control group(each n=65)based on whether IOUS was used during operation.The general information,resection related indicators and postoperative complications were compared between groups,and the application value of IOUS was analyzed.Results In IOUS group,the tumor depth was greater,while the intraoperative bleeding,duration of hospitalization and operation-duration were all less than those in control group(all P<0.05).Postoperative complications occurred in 28 cases of IOUS group and 29 cases of control group,and the incidence of subdural effusion in IOUS group was lower than that in control group(P<0.05).Conclusion IOUS could effectively reduce intraoperative bleeding,duration of hospitalization,operation-duration and incidence of postoperative subdural effusion in microsurgical resection of primary supratentorial glioblastoma.
7.Intraoperative ultrasound radiomics for predicting isocitrate dehydrogenase 1(IDH1)mutation of high-grade glioma
Huiyu MA ; Wei ZHANG ; Chao HOU ; Linggang CHENG ; Wenkai ZHANG ; Lizhi YANG ; Wen HE
Chinese Journal of Medical Imaging Technology 2025;41(4):569-572
Objective To investigate the value of intraoperative ultrasound radiomics for predicting isocitrate dehydrogenase 1(IDH1)mutation of high-grade glioma.Methods Ninety-five patients with high-grade glioma(WHO grade Ⅲ and Ⅳ)who underwent craniotomy glioma resection and ultrasound assisted tumor localization during operation and then confirmed by pathology were retrospectively enrolled.The patients were divided into training set(n=66,including 24 IDH1 mutation type and 42 IDH1 wild type)and validation set(n=29,including 11 IDH1 mutation type and 18 IDH1 wild type)at the ratio of 7∶3.Based on intraoperative ultrasound,radiomics features were extracted,the best ones were screened,and a radiomics model was established for predicting IDH1 mutation of high-grade glioma using random forest algorithm.Receiver operating characteristic(ROC)curve was plotted,the area under the curve(AUC)was calculated to evaluate the predictive efficacy of the model,and decision curve analysis(DCA)was used to evaluate the clinical value of the model.Results A total of 851 radiomics features were extracted based on intraoperative ultrasound,and finally 5 best ones were screened out to construct a radiomics model.The AUC of the radiomics model for predicting IDH1 mutation of high-grade glioma in training and validation sets was 0.902 and 0.707,respectively,with no significant difference(P=0.097).DCA maps showed that the clinical net benefit of the radiomics model was high.Conclusion Intraoperative ultrasound radiomics could effectively predict IDH1 mutation of high-grade glioma.
8.Value of preoperative ultrasound combined with NLR,Ctn and CEA levels in predicting cervical lymph node metastasis of papillary thyroid carcinoma
Shanshan YAN ; Huiyu ZHANG ; Jun GU ; Kan ZHANG ; Yu ZHAO
The Journal of Practical Medicine 2025;41(18):2859-2864
Objective To investigate the predictive value of preoperative ultrasound in combination with neutrophil-to-lymphocyte ratio(NLR),calcitonin(Ctn),and carcinoembryonic antigen(CEA)for cervical lymph node metastasis in patients with papillary thyroid carcinoma.Methods A total of 103 patients diagnosed with papillary thyroid carcinoma(PTC)who were admitted to the hospital between October 2021 and October 2024 were selected as the case group.Among them,34 patients with cervical lymph node metastasis confirmed by surgical and pathological examination were assigned to the metastatic group,and 69 patients without cervical lymph node metas-tasis were assigned to the non-metastatic group.Additionally,103 patients with benign thyroid nodules admitted during the same period were enrolled as the control group.Clinical data,ultrasonographic features,and serum levels of NLR,Ctn,and CEA were compared between the metastatic and non-metastatic groups.The predictive value of ultrasonographic features and the combined detection of NLR,Ctn,and CEA levels for cervical lymph node metastasis in PTC was evaluated using receiver operating characteristic(ROC)curve analysis.Results Compared to the control group,the case group exhibited a higher proportion of patients with microcalcification and grade 3 blood flow,as well as elevated levels of NLR,Ctn,and CEA(P<0.05).Similarly,compared to the non-metastatic group,the metastatic group showed a higher proportion of patients with microcalcification and grade 3 blood flow,along with increased levels of NLR,Ctn,and CEA(P<0.05).The metastatic group tested positive,whereas the non-metastatic group tested negative.The area under the curve(AUC)for ultrasound features(micro-calcification,blood flow classification)combined with NLR,Ctn,and CEA levels in diagnosing cervical lymph node metastasis in papillary thyroid carcinoma was higher than that of individual indicators(P<0.05).Conclusions Preoperative ultrasound combined with the assessment of NLR,Ctn,and CEA levels demonstrates significant predictive value for cervical lymph node metastasis in papillary thyroid carcinoma.
9.Mechanisms and Prospects of EGCG in the Prevention and Treatment of Atheroscle-rosis
Xueqi DAI ; Guanjie ZHANG ; Jing LI ; Huiyu LONG ; Hui SUN
Chinese Journal of Arteriosclerosis 2025;33(9):803-809
Atherosclerosis(As)is a chronic inflammatory vascular disease and serves as the pathological basis for various cardiovascular diseases.Epigallocatechin-3-O-gallate(EGCG)is the most abundant and biologically active com-pound in green tea,possessing multiple pharmacological effects such as anti-inflammatory,antiviral,and antioxidative stress properties.In recent years,studies have found that EGCG can regulate lipid metabolism,improve vascular endo-thelial function,and enhance plaque stability,demonstrating its potential value in combating As.This article provides a systematic review of the mechanisms underlying EGCG's anti-As effects,aiming to offer multi-faceted theoretical support for its further research and clinical application in the prevention and treatment of As.
10.Effect of C1q tumor necrosis factor-related protein 3 on reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells by Sendai virus vector overexpressing Gata4,Mef2c,and Tbx5
Yanbin SONG ; Yunqing ZHANG ; Huiyu LIU ; Junming CHEN
Chinese Journal of Comparative Medicine 2025;35(8):14-28
Objective To investigate the efficiency and mechanism of C1q tumor necrosis factor-related protein 3(CTRP3)on reprogramming of cardiac fibroblasts(CFs)into induced cardiomyocyte-like cells(iCMs)by Sendai virus(SeV)vector overexpressing Gata4,Mef2c and Tbx5(SeVGMT).Methods CFs were divided into Control,NC-Lv,CTRP3-Lv,NC-sh,and CTRP3-sh groups.NC-Lv,CTRP3-Lv,NC-sh,and CTRP3-sh were transfected into CFs using Lipofectamine 3000 reagent for 48 hours.Lipofectamine 3000 reagent was then mixed with SeVGMT and incubated at room temperature for 48 hours,the culture medium was then replaced,and cells were cultured for 21 days.Cell morphology was observed under a microscope at 0,3,7,14,and 21 days.Expression levels of the myocardial-specific proteins α-myosin heavy chain(α-MHC),α-actin,cardiac troponin T(cTnT),connexin 43(Cx43),cardiac muscle α-actin(Actc1),and myosin heavy chain 6(Myh6)were detected at different time points by immunofluorescence,quantitative reverse transcription-polymerase chain reaction,and Western blot,and the proportions of beating cells at different time points were calculated.Results The relative fluorescence intensity and mRNA and protein levels of α-MHC,α-actin,cTnT,Cx43,Actc1,and Myh6 in CFs in each group increased with increasing culture time(P<0.05),with significantly higher expression levels of myocardial-specific proteins at 14 days of culture than at 7 days(P<0.05).The relative fluorescence intensities and mRNA and protein levels of α-MHC,α-actin,cTnT,Cx43,Actc1,and Myh6 in CFs at 3,7,14,and 21 days of culture were significantly increased in the CTRP3-Lv group compared with the NC-Lv group(P<0.05),but were significantly decreased in CFs in the CTRP3-sh group compared with the NC-sh group(P<0.05).Beating cells appeared in CFs in each group at 7 days of culture.The proportion of beating cells in each group increased with increasing culture time(P<0.05),and the proportion was significantly higher at 14 days than at 7 days(P<0.05).The proportion of beating cells among CFs was increased in the CTRP3-Lv group at 7,14,and 21 days of culture compared with the NC-Lv group(P<0.05),while the proportion of beating cells in the CTRP3-sh group was decreased compared with the NC-sh group(P<0.05).Conclusions CTRP3 can enhance SeVGMT reprogramming of CFs into iCMs.

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