1.Extraction,Separation and Hypoglycemic Activity Analysis of Polysaccharides from Brassica rapa
Mengyu HOU ; Ruina XU ; Qingsong LI ; Shaoxuan LI ; Xinying MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):219-228
ObjectiveTo optimize the extraction method for polysaccharides from turnip(Brassica rapa), and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction(BP-1) and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index, a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio, extraction time and extraction temperature, in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry(UV), gas chromatography-mass spectrometry(GC-MS), Congo red assay, and Fourier-transform infrared spectroscopy(FT-IR) to determine purity, monosaccharide composition, triple-helix structure, and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy(SEM) and atomic force microscopy(AFM). Zebrafish were divided into the blank group(adding E3 medium), and BP-1-1, BP-1-10, BP-1-50, BP-1-200, BP-1-1 000 groups(adding BP-1 solutions at concentrations of 1, 10, 50, 200, 1 000 mg·L-1, respectively), and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype, survival rate, malformation rate, and heart rate. Experimental animals were randomly divided into the blank group, model group, BP-1-10 group(10 mg·L-1), BP-1-50 group(50 mg·L-1), and BP-1-200 group(200 mg·L-1). The blank group was cultured in E3 medium, the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization(dpf) zebrafish embryos using 10 g·L-1 of glucose combined with 500 µmol·L-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution, while the blank and model groups received an equal volume of saline. Glucose and insulin(INS) levels were measured using enzyme-linked immunosorbent assay(ELISA) kits, the effects on the liver were observed by hematoxylin-eosin(HE) histopathological sections. The mRNA expression levels of glucagon(Glucagon), insulin(Insa), and phosphoenolpyruvate carboxykinase 1(PCK1) were detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). ResultsThe optimized extraction conditions were determined as follows:solid-to-liquid ratio of 1∶40(g·mL-1), extraction time of 66 min, and extraction temperature of 79 ℃. Under these conditions, the yield of turnip polysaccharides was (10.34±0.96)%. UV analysis indicated that BP-1 contained no proteins or nucleic acids, GC-MS analysis revealed that BP-1 consisted of six monosaccharides(arabinose, rhamnose, ribose, mannose, galactose and glucose). Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure, FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids, SEM analysis revealed an irregular flaky structure with a flat and smooth surface, AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L-1. Pharmacodynamic results showed that, compared with the blank group, the model group exhibited significantly increased glucose levels and significantly decreased INS levels(P<0.01). Compared with the model group, the BP-1-50 group significantly reduced glucose levels and increased INS levels(P<0.05). Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal, with hepatocytes appearing plump. Real-time PCR results showed that, compared with the blank group, the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1, and significantly downregulated mRNA expression of Insa(P<0.01). Compared with the model group, the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1, and significantly upregulated mRNA expression of Insa(P<0.01). ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate, is simple to operate, has low costs, making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides, contains α-glycosidic bonds and uronic acids, exhibits hypoglycemic activity, and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.
2.Extraction,Separation and Hypoglycemic Activity Analysis of Polysaccharides from Brassica rapa
Mengyu HOU ; Ruina XU ; Qingsong LI ; Shaoxuan LI ; Xinying MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):219-228
ObjectiveTo optimize the extraction method for polysaccharides from turnip(Brassica rapa), and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction(BP-1) and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index, a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio, extraction time and extraction temperature, in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry(UV), gas chromatography-mass spectrometry(GC-MS), Congo red assay, and Fourier-transform infrared spectroscopy(FT-IR) to determine purity, monosaccharide composition, triple-helix structure, and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy(SEM) and atomic force microscopy(AFM). Zebrafish were divided into the blank group(adding E3 medium), and BP-1-1, BP-1-10, BP-1-50, BP-1-200, BP-1-1 000 groups(adding BP-1 solutions at concentrations of 1, 10, 50, 200, 1 000 mg·L-1, respectively), and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype, survival rate, malformation rate, and heart rate. Experimental animals were randomly divided into the blank group, model group, BP-1-10 group(10 mg·L-1), BP-1-50 group(50 mg·L-1), and BP-1-200 group(200 mg·L-1). The blank group was cultured in E3 medium, the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization(dpf) zebrafish embryos using 10 g·L-1 of glucose combined with 500 µmol·L-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution, while the blank and model groups received an equal volume of saline. Glucose and insulin(INS) levels were measured using enzyme-linked immunosorbent assay(ELISA) kits, the effects on the liver were observed by hematoxylin-eosin(HE) histopathological sections. The mRNA expression levels of glucagon(Glucagon), insulin(Insa), and phosphoenolpyruvate carboxykinase 1(PCK1) were detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). ResultsThe optimized extraction conditions were determined as follows:solid-to-liquid ratio of 1∶40(g·mL-1), extraction time of 66 min, and extraction temperature of 79 ℃. Under these conditions, the yield of turnip polysaccharides was (10.34±0.96)%. UV analysis indicated that BP-1 contained no proteins or nucleic acids, GC-MS analysis revealed that BP-1 consisted of six monosaccharides(arabinose, rhamnose, ribose, mannose, galactose and glucose). Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure, FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids, SEM analysis revealed an irregular flaky structure with a flat and smooth surface, AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L-1. Pharmacodynamic results showed that, compared with the blank group, the model group exhibited significantly increased glucose levels and significantly decreased INS levels(P<0.01). Compared with the model group, the BP-1-50 group significantly reduced glucose levels and increased INS levels(P<0.05). Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal, with hepatocytes appearing plump. Real-time PCR results showed that, compared with the blank group, the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1, and significantly downregulated mRNA expression of Insa(P<0.01). Compared with the model group, the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1, and significantly upregulated mRNA expression of Insa(P<0.01). ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate, is simple to operate, has low costs, making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides, contains α-glycosidic bonds and uronic acids, exhibits hypoglycemic activity, and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.
3.Study on the mechanism of Brassica rapa polysaccharide in improving alcoholic liver injury of mice based on intestinal microbiota and metabolomics
Xinying MA ; Ruina XU ; Shaoxuan LI ; Ruiyin YE ; Yuexing MA ; Yaohui YE
China Pharmacy 2025;36(16):2005-2011
OBJECTIVE To investigate the effects of Brassica rapa polysaccharide (BRP) on the Toll-like receptor 4 (TLR4)/ myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB), AMP-activated protein kinase (AMPK)/sterol regulatory element-binding protein-1c (SREBP-1c) pathways, intestinal microbiota and liver metabolism of mice with alcoholic liver injury, and preliminarily elucidate its mechanism for improving alcoholic liver injury. METHODS Seventy-two mice were randomly divided into blank group (normal saline), model group (normal saline), bifendate group (positive control, 300 mg/kg) and BRP low-, medium- and high-dose groups (75, 150 and 300 mg/kg). They were given relevant medicine intragastrically, once a day, for consecutive 9 d. After the last administration, mice in all groups except the blank group were gavaged with white liquor to establish an alcoholic liver injury model. The levels of alanine aminotransferase and aspartate aminotransferase in serum, total cholesterol, triglycerides, low-density lipoprotein cholesterol, interleukin-6, interleukin-1β, tumor necrosis factor- α and lipopolysaccharide, as well as protein expressions of TLR4, MyD88, NF-κB p65, phosphorylated NF-κB p65 (p-NF-κB p65), AMPK, phosphorylated AMPK (p-AMPK), and SREBP-1c were all detected; pathological morphological changes of liver tissue and colon were observed. 16S rRNA was used to detect the changes of intestinal microbiota in mice, and metabolomics 2022B02058) technology was used to detect the changes of liver metabolites. RESULTS Compared with model group, the above biochemical indicators and the protein expressions of TLR4, MyD88, p-NF-κB p65, and SREBP-1c in liver tissues were all significantly decreased (P<0.05 or P<0.01), while the protein expression of p-AMPK was significantly increased (P<0.05 or P<0.01). Pathological damage to liver and colon tissues was significantly improved. Medium dose of BRP could increase the relative abundance of Akkermansia, norank_f_Muribaculaceae and Lachnospiraceae_NK4A136_group in the intestinal contents of mice to a certain extent, and decrease the relative abundance of Lactobacillus and Escherichia-Shigella. A total of 9 differential metabolites were identified by metabolomics, including homogentisic acid, myristyl lysophosphatidylcholine, which were involved in pathways such as tyrosine metabolism. CONCLUSIONS BRP can regulate the relative abundance of beneficial flora, reduce the relative abundance of harmful flora, improve the structure of intestinal colonies, reduce the entry of pro-inflammatory mediator lipopolysaccharides into liver tissue, affect metabolic pathways such as tyrosine metabolism and the expression of TLR4/MyD88/NF- κB and AMPK/SREBP-1c signaling pathways in the liver, and ultimately improve alcoholic liver injury.
4.Research progress of hypoglycemic agents on circulating PCSK9 concentration
Weijie FAN ; Shanghai LI ; Ruina HUANG ; Xinbin ZHANG ; Wantai WU ; Weijun LIANG
Clinical Medicine of China 2025;41(1):71-75
Long-term glucose metabolism disorders in patient with diabetes is often accompanied by dyslipidemia, especially the elevated level of low-density lipoprotein cholesterol (LDL-C). It is an important risk factor for the development of atherosclerotic cardiovascular disease (ASCVD). Recent studies at home and abroad have also found that diabetes can lead to increased levels of proprotein convertase subtilisin/kexintype 9 (PCSK9). In view of the profound influence of PCSK9 overexpression on low density lipoprotein cholesterol level, cardiovascular risk and atherosclerosis, it is of great significance to explore whether hypoglycemic drugs can reduce the level of PCSK9 in the treatment of diabetes. We summarized the results of the study on the effects of various hypoglycemic drugs on circulating PCSK9 concentration in order to provide ideas for the optimal treatment of diabetes.
5.Evaluation of the therapeutic effect and influencing factors of personalized exercise on patients with type 2 diabetes mellitus by dynamic blood glucose profile
Lilan SHEN ; Fulian LI ; Juanqin LYU ; Ruina ZHANG ; Peiyun ZENG
Chinese Journal of Diabetes 2025;33(7):497-500
Objective To explore the therapeutic effect and influencing factors of individualized exercise under continuous glucose profile in patients with typ 2 diabetes mellitus(T2DM).Methods A total of 110 patients with T2DM who were treated in the Department of Endocrinology of our hospital from December 2021 to July 2022 were randomly divided into conventional group and personalized group,with 55 cases in each group.The conventional group was given routine health exercise guidance,the personalized group was given a personalized exercise program.Repeated measurement analysis of variance was used to evaluate the improvement of blood glucose related indicators in the two groups at 3 and 6 months after intervention.Logistic regression was used to analyze the influencing factors of blood glucose(BG)control after 6 months of intervention.Results After 3 and 6 months of intervention,FBG,2 hBG,hemoglobin A1c(HbA1c),mean blood glucose(MBG)and time above target range(TAR)in the personalized group and conventional group were all lower than those before intervention(P<0.05).Time in range(TIR)of glucose within 24 h was higher than that before intervention(P<0.05).After 3 and 6 months of intervention,FBG,2 hBG,HbA1c,MBG and TAR in the personalized group were lower than those in the conventional group at the same time point(P<0.05),and TIR was higher than that in the conventional group at the same time point(P<0.05).After 6 months of intervention,the BG control rate in the personalized group was higher than that in the conventional group(P<0.05).Logistic regression analysis showed that the DM duration,exercise intervention methods and TIR were the influencing factors of BG control.Conclusions Exercise guidance is an effective intervention method to control diabetes.Compared with conventional exercise guidance,personalized exercise program has a better effect on controlling patients'BG level,and has a better effect on patients with short course of disease.
6.Guide on Methodological Standards in Pharmacoepidemiology(2nd edition)and their series interpretation(9):research report standards and results visualization
Jingru CHENG ; Ruina CHEN ; Jiarui LI ; Shaowen TANG ; Feng SUN ; Siyan ZHAN
Chinese Journal of Pharmacoepidemiology 2025;34(9):1004-1016
Standardized research reporting is crucial for the translation of pharmacoepidemiology research findings,and visual reporting can significantly enhance the clarity,understandability,and transparency of research results.Based on the Guide on Methodological Standards in Pharmacoepidemiology(2nd edition),this article systematically explains the key points for writing each component of a research report(including title,abstract,introduction,research methods,research results,discussion and conclusions,acknowledgments,conflict of interest statement,and references).This article also summarizes recognized international and domestic standards for pharmacoepidemiology research reporting,providing a reference for researchers.Furthermore,real-world cases will be used to demonstrate common forms of visualized reports and their interpretation methods.Finally,it further explores strategies for communicating research results.This study aims to provide pharmacoepidemiology researchers with detailed guidance on visually presenting research results and writing high-quality research reports,thereby enhancing the integrity and impact of their research.
7.Guide on Methodological Standards in Pharmacoepidemiology(2nd edition)and their series interpretation(9):research report standards and results visualization
Jingru CHENG ; Ruina CHEN ; Jiarui LI ; Shaowen TANG ; Feng SUN ; Siyan ZHAN
Chinese Journal of Pharmacoepidemiology 2025;34(9):1004-1016
Standardized research reporting is crucial for the translation of pharmacoepidemiology research findings,and visual reporting can significantly enhance the clarity,understandability,and transparency of research results.Based on the Guide on Methodological Standards in Pharmacoepidemiology(2nd edition),this article systematically explains the key points for writing each component of a research report(including title,abstract,introduction,research methods,research results,discussion and conclusions,acknowledgments,conflict of interest statement,and references).This article also summarizes recognized international and domestic standards for pharmacoepidemiology research reporting,providing a reference for researchers.Furthermore,real-world cases will be used to demonstrate common forms of visualized reports and their interpretation methods.Finally,it further explores strategies for communicating research results.This study aims to provide pharmacoepidemiology researchers with detailed guidance on visually presenting research results and writing high-quality research reports,thereby enhancing the integrity and impact of their research.
8.Evaluation of the therapeutic effect and influencing factors of personalized exercise on patients with type 2 diabetes mellitus by dynamic blood glucose profile
Lilan SHEN ; Fulian LI ; Juanqin LYU ; Ruina ZHANG ; Peiyun ZENG
Chinese Journal of Diabetes 2025;33(7):497-500
Objective To explore the therapeutic effect and influencing factors of individualized exercise under continuous glucose profile in patients with typ 2 diabetes mellitus(T2DM).Methods A total of 110 patients with T2DM who were treated in the Department of Endocrinology of our hospital from December 2021 to July 2022 were randomly divided into conventional group and personalized group,with 55 cases in each group.The conventional group was given routine health exercise guidance,the personalized group was given a personalized exercise program.Repeated measurement analysis of variance was used to evaluate the improvement of blood glucose related indicators in the two groups at 3 and 6 months after intervention.Logistic regression was used to analyze the influencing factors of blood glucose(BG)control after 6 months of intervention.Results After 3 and 6 months of intervention,FBG,2 hBG,hemoglobin A1c(HbA1c),mean blood glucose(MBG)and time above target range(TAR)in the personalized group and conventional group were all lower than those before intervention(P<0.05).Time in range(TIR)of glucose within 24 h was higher than that before intervention(P<0.05).After 3 and 6 months of intervention,FBG,2 hBG,HbA1c,MBG and TAR in the personalized group were lower than those in the conventional group at the same time point(P<0.05),and TIR was higher than that in the conventional group at the same time point(P<0.05).After 6 months of intervention,the BG control rate in the personalized group was higher than that in the conventional group(P<0.05).Logistic regression analysis showed that the DM duration,exercise intervention methods and TIR were the influencing factors of BG control.Conclusions Exercise guidance is an effective intervention method to control diabetes.Compared with conventional exercise guidance,personalized exercise program has a better effect on controlling patients'BG level,and has a better effect on patients with short course of disease.
9.Research progress of hypoglycemic agents on circulating PCSK9 concentration
Weijie FAN ; Shanghai LI ; Ruina HUANG ; Xinbin ZHANG ; Wantai WU ; Weijun LIANG
Clinical Medicine of China 2025;41(1):71-75
Long-term glucose metabolism disorders in patient with diabetes is often accompanied by dyslipidemia, especially the elevated level of low-density lipoprotein cholesterol (LDL-C). It is an important risk factor for the development of atherosclerotic cardiovascular disease (ASCVD). Recent studies at home and abroad have also found that diabetes can lead to increased levels of proprotein convertase subtilisin/kexintype 9 (PCSK9). In view of the profound influence of PCSK9 overexpression on low density lipoprotein cholesterol level, cardiovascular risk and atherosclerosis, it is of great significance to explore whether hypoglycemic drugs can reduce the level of PCSK9 in the treatment of diabetes. We summarized the results of the study on the effects of various hypoglycemic drugs on circulating PCSK9 concentration in order to provide ideas for the optimal treatment of diabetes.
10.Intervention of traditional Chinese patent medicine based on syndrome differentiation in female patients after percutaneous coronary intervention due to acute coronary syndrome: a nationwide multicenter prospective cohort study
Ruina BAI ; Feng GU ; Yajie CAI ; Song SHENG ; Qiaoning YANG ; Ruixi XI ; Lizhi LI ; Dazhuo SHI
Chinese Journal of Cardiology 2024;52(9):1073-1080
Objective:To evaluate the efficacy and safety of discriminative application of Chinese patent medicines in female patients after percutaneous coronary intervention (PCI) due to acute coronary syndrome (ACS).Methods:The study population was from the Chinese Patent Medicine (CPM) trial. CPM trial was a multicenter prospective cohort study, which enrolled patients from 40 centers in mainland China between February 2012 and December 2015, with the discriminative use of Chinese patent medicines as the exposure factor. Female patients with ACS after PCI who completed 36-month follow-up were included in this analysis, and were divided into a conventional treatment group (using conventional western medicine recommended by the guidelines) and a group with the discriminative use of proprietary Chinese medicines (on the basis of conventional western medicine treatment, discriminative use of Qishen Yiqi dropping pills for Qi deficiency and blood stasis syndrome, Guanxin Danshen dropping pills for blood stasis syndrome, and Danlou tablets for phlegm and blood stasis syndrome combined with the conventional western medicine). The primary endpoint event was a composite endpoint event including cardiovascular death, nonfatal myocardial infarction, and emergency revascularization surgery. Secondary endpoint events were composite endpoint events including readmission for ACS, heart failure, stroke, and other thrombotic events. Adverse events were collected. Cox proportional risk model was used to assess the effect of discriminatory application of Chinese patent medicine on endpoint events, and sensitivity analysis was performed by comparing the results with propensity score matching analysis.Results:A total of 748 female ACS post-PCI patients were included in the analysis, aged (63.2±8.3) years. There were 370 patients in the group of discriminative application of Chinese patent medicines and 378 patients in the conventional treatment group. There were 37 cases (10.0%) and 58 cases (15.3%) of primary endpoint events in the discriminatory application of Chinese patent medicines group and the conventional treatment group, respectively. Cox analysis showed that the risk of primary endpoint in the discriminatory application of Chinese patent medicines group was lower than that in the conventional treatment group after adjusting for confounding factors (adjusted HR=0.62, 95% CI 0.40-0.96, P=0.031). There were 38 (10.3%) and 57 (15.1%) cases of secondary endpoint events in the two groups, respectively. Cox regression analysis showed that the risk of secondary endpoint events in the discriminatory application of Chinese patent medicine group was lower than that in the conventional treatment group after adjusting for confounders (adjusted HR=0.56, 95% CI 0.37-0.87, P=0.001). The results of propensity score matching analysis also showed that Chinese patent medicines based on discriminatory application could reduce the risk of primary endpoint ( HR=0.62,95% CI 0.40-0.97 ,P=0.033) and second endpoint ( HR=0.56, 95% CI 0.37-0.87, P=0.009) significantly. There was no significant difference in adverse events between the two groups (12.4% (46/370) vs. 10.3% (39/378), P=0.362). Conclusion:On the basis of conventional western medicine treatment, discriminatory application of Chinese patent medicines can reduce the risk of endpoints in female patients after PCI due to ACS without significant adverse effects.

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