1.Development of DUS Test Guidelines for New Pinellia ternata
Xinyao LI ; Mingxing WANG ; Bingbing LIAO ; Changjie CHEN ; Xiufu WAN ; Lanping GUO ; Yuhuan MIAO ; Dahui LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):225-233
Pinellia ternata, belonging to the Pinellia genus within the Araceae family, is a medicinal plant due to its tubers. There are severe issues with unclear germplasm and mixed varieties in its cultivation, necessitating urgent new variety protection efforts. The distinctness, uniformity, and stability (DUS) testing of the plant variety is the basis for protecting new plant varieties, and the DUS test guidelines are the technical basis for DUS testing. To develop the DUS test guidelines for P. ternata, agronomic traits of 229 germplasm of P. ternata were observed and measured during its two growth stages over the years, and each character was graded and described. A total of 38 traits were selected as the test traits of the DUS test guideline for P. ternata. There were three plant traits, 19 leaf traits, six flower traits, two fruit traits, two tuber traits, five bulbil traits, and one ploidy trait. These traits could be divided into 22 quality characters, 12 quantitative characters, and four pseudo-quantitative characters, as well as seven groups, including plants, leaves, flowers, fruit, tubers, bulbils, and ploidy. By searching for standard traits, 10 standard varieties were ultimately determined. Preparing these guidelines will have great significance for reviewing and protecting P. ternata varieties, safeguarding breeders' rights, and promoting the development of the P. ternata industry.
2.Apoptosis Regulation by Buzhong Yiqitang via PERK/eIF2α/ATF4/CHOP Pathway to Enhance Pulmonary Surfactant-associated Protein C Expression and Ameliorate Lung Injury in CIH Mice
Luyao ZHANG ; Yangjing WANG ; Bingbing LIU ; Jieru LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):19-27
ObjectiveTo investigate the effects of Buzhong Yiqitang on the abnormal expression of pulmonary surfactant-associated protein C (SFTPC) and lung injury induced by chronic intermittent hypoxia (CIH) and the mechanism of action. MethodsForty healthy adult male SPF-grade C57BL/6 mice were randomly allocated into five experimental groups: a normoxia group, a CIH group, and low-, medium-, and high-dose Buzhong Yiqitang groups, with eight mice in each group. During the modeling, mice in the normoxia group were housed under standard oxygen concentrations, while the CIH and all Buzhong Yiqitang groups were placed in a hypoxic chamber for 8 h daily over 35 d. Prior to each chamber session, mice in the low-, medium-, and high-dose Buzhong Yiqitang groups were administered decoctions by gavage at corresponding doses (8.1, 16.2, 32.4 g·kg-1·d-1 of crude drug, respectively), while those in normoxia and CIH groups received an equivalent volume of saline by gavage. The general conditions of the mice were recorded before and after the experiment. Pulmonary function was assessed using a non-invasive detection system. Serum SFTPC levels were measured using enzyme-linked immunosorbent assay (ELISA). Histopathological changes in lung tissue were evaluated using hematoxylin-eosin (HE) staining. Apoptosis in lung tissue was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Protein expression of SFTPC, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), protein kinase R-like endoplasmic reticulum kinase (PERK), phosphorylated PERK (p-PERK), eukaryotic initiation factor 2α (eIF2α), phosphorylated eIF2α (p-eIF2α), activating transcript factor 4 (ATF4), and CCAAT/enhancer-binding protein homologous protein (CHOP) in lung tissue was analyzed by Western blot. Immunofluorescence staining was employed to assess the expression of SFTPC and CHOP proteins in lung tissue. ResultsCompared to those in the normoxia group, mice in the CIH group showed significantly impaired pulmonary function and increased histopathological lung injury scores (P<0.05, P<0.01). Serum SFTPC levels increased, while SFTPC expression in lung tissue was reduced (P<0.05, P<0.01). The rate of apoptotic cells in lung tissue increased, and the expression of endoplasmic reticulum stress markers p-PERK, p-eIF2α, ATF4, and CHOP was upregulated (P<0.05, P<0.01). Compared with the CIH group, Buzhong Yiqitang intervention improved pulmonary function indicators and decreased the histopathological lung injury scores (P<0.05, P<0.01). Serum SFTPC levels were decreased, and lung tissue SFTPC expression was recovered (P<0.05, P<0.01). The apoptotic rate of lung tissue cells was significantly reduced, with downregulation of pro-apoptotic Bax and upregulation of anti-apoptotic Bcl-2 expression (P<0.05, P<0.01). Activation and expression of p-PERK, p-eIF2α, ATF4, and CHOP were also decreased (P<0.05, P<0.01). ConclusionBuzhong Yiqitang can alleviate lung injury and improve pulmonary function by reducing lung cell apoptosis and enhancing alveolar surfactant secretion, which may be related to the modulation of the PERK/eIF2α/ATF4/CHOP signaling pathway.
3.A proteomic landscape of pharmacologic perturbations for functional relevance
Zhiwei LIU ; Shangwen JIANG ; Bingbing HAO ; Shuyu XIE ; Yingluo LIU ; Yuqi HUANG ; Heng XU ; Cheng LUO ; Min HUANG ; Minjia TAN ; Jun-Yu XU
Journal of Pharmaceutical Analysis 2024;14(1):128-139
Pharmacological perturbation studies based on protein-level signatures are fundamental for drug dis-covery.In the present study,we used a mass spectrometry(MS)-based proteomic platform to profile the whole proteome of the breast cancer MCF7 cell line under stress induced by 78 bioactive compounds.The integrated analysis of perturbed signal abundance revealed the connectivity between phenotypic behaviors and molecular features in cancer cells.Our data showed functional relevance in exploring the novel pharmacological activity of phenolic xanthohumol,as well as the noncanonical targets of clinically approved tamoxifen,lovastatin,and their derivatives.Furthermore,the rational design of synergistic inhibition using a combination of histone methyltransferase and topoisomerase was identified based on their complementary drug fingerprints.This study provides rich resources for the proteomic landscape of drug responses for precision therapeutic medicine.
4.Influence of different compressed sensing factors on susceptibility weighted imaging for displaying cerebral medullary vein
Jiajun CAO ; Jing YANG ; Yukun ZHANG ; Na LIU ; Bingbing GAO ; Yangyingqiu LIU ; Qingwei SONG ; Yanwei MIAO
Chinese Journal of Interventional Imaging and Therapy 2024;21(3):139-144
Objective To observe the influence of different acceleration factors(AF)on compressed sensing susceptibility weighted imaging(CS-SWI)for cerebral medullary veins of healthy people,and to screen the best AF.Methods Forty healthy volunteers were prospectively enrolled.Axial brain SWI images were obtained with CS technique under different AF(AF0,CS2,CS4,CS6,CS8 and CS10),and the phase value(PV)and standard deviation(SD)of bilateral septal vein(SV),internal cerebral vein(ICV),thalamus vein(TV),basal vein(BV)and dentate nucleus vein(DNV)were measured.Taken PV and SD of parietal white matter as controls,the signal-to-noise ratio(SNR)and contrast-to-noise ratio(CNR)of venous images were calculated.Then the original images were reconstructed with minimum intensity projection,and the subjective quality scoring of reconstructed images were performed using a 5-point scale.SNR,CNR,PV and quality score were compared among images under different AF,and the best AF,i.e.with the best performances for displaying and quantitatively analyzing cerebral medullary veins in healthy people was obtained.Results Compared with those acquired with AF0,SNR and CNR of all cerebral medullary veins acquired with CS6,CS8 and CS10 were significantly different(all adjusted P<0.05).Meanwhile,significant differences of PV in bilateral SV and right TV were found among CS6,CS8 and CS10,also in bilateral ICV,left TV and bilateral BV between CS8 and CS10(all adjusted P<0.05).Conclusion Excessive AF might decrease image quality of CS-SWI for cerebral medullary veins.CS4 was the best AF for displaying and quantitatively analyzing cerebral medullary veins in healthy people.
5.Exploration on the Mechanism of Hydroxyl Safflower Flavin A in the Treatment of Sepsis-induced Liver Injury Based on Metabolomics and Network Pharmacology
Shifan YAN ; Bingbing PAN ; Ting YU ; Changmiao HOU ; Yu JIANG ; Fang CHEN ; Jingjing WANG ; Yanjuan LIU ; Yimin ZHU
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(2):130-137
Objective To explore the mechanism of hydroxyl safflower flavin A(HSYA)in the treatment of sepsis-induced liver injury by using metabolomics and network pharmacology.Methods A total of 50 male C57BL/6 mice were randomly divided into sham-operation group(10 mice),sepsis group(20 mice)and HSYA group(20 mice).Cecal ligation and puncture was conducted to establish the sepsis-induced liver injury mouse model.The mice in HSYA group were subcutaneously injected with HSYA after 2 hours of modeling.The content of serum inflammatory factors and liver function were detected,and the pathological changes of liver tissue were observed with HE staining,UPLC-Q-TOF-MS metabolomics was used to analyze liver tissue,screening for differential metabolites using multivariate statistical methods,network pharmacology was used to predict potential targets for HSYA treatment of sepsis-induced liver injury,and conduct GO and KEGG pathway enrichment analysis on potential targets,Metabo Analyst 5.0 database was used to match differential metabolites and potential targets between the model group and HSYA group,a targets metabolite-metabolism pathway network was constructed.AutoDock Vina software was used to perform molecular docking between HSYA and core genes,and finally RT-qPCR was used to verify the expression of core genes.Results HSYA can reduce the contents of IL-6,IL-1β and TNF-α in serum,restore liver function,and alleviate the morphological alternation in liver induced by sepsis.A total of 26 differential metabolites identified by metabolomics were screened out,including flufenamic acid,cryptolepine,opthalmic acid,fenpropathrin etc.,which were mainly involved in 5 metabolic pathways such as biosynthesis of unsaturated fatty acids and alpha-linolenic acid metabolism.Network pharmacology identified 81 potential targets,2 735 items enriched in GO and 124 signaling pathways enriched in KEGG;a total of 5 differential metabolites were matched for joint analysis,corresponding to 14 targets including IL1B,STAT3,PTGS2,TP53,etc.,involved in the regulation of metabolic disorders in sepsis-induced liver injury by HSYA.Molecular docking results showed that HSYA had good binding activity to IL1B,STAT3,PTGS2 and TP53 targets.RT-qPCR results showed that HSYA could inhibit the expressions of IL1B,STAT3 and PTGS2 in liver tissue.Conclusions HSYA may inhibit the release of inflammatory cytokines,maintain metabolic homeostasis,and alleviate sepsis-induced liver injury through modulating the expressions of IL1B,STAT3,and PTGS2.
6.Ultrasonic evaluation of fetal cerebral sulci and gyrus development in pregnant women with gestational diabetes mellitus
Xiaolin ZHANG ; Zhaoling ZHU ; Ruili WANG ; Yuan GAO ; Bingbing LIU ; Liangjie GUO ; Jianjun YUAN ; Jingge ZHAO
Chinese Journal of Ultrasonography 2024;33(1):36-41
Objective:To evaluate the development of fetal cerebral sulci and gyrus and the blood perfusion in pregnant women with gestational diabetes mellitus(GDM) by ultrasound.Methods:A total of 1 540 pregnant women with 28-34 weeks of pregnancy who underwent systematic screening in Henan Provincial People′s Hospital from January 2022 to October 2022 were prospectively selected, 100 pregnant women with GDM were selected as the GDM group. According to the effect of blood glucose control, the GDM group was divided into 2 groups: the satisfied control group (GDM group 1), and the dissatisfied control group (GDM group 2), with 50 cases in each group. At the same period, 50 healthy pregnant women at 28-34 weeks of gestation were enrolled as the control group. The differences of the sylvian fissure, parietooccipital sulci, calcarine sulci and cinguli sulci among the 3 groups were statistically analyzed. And the correlations between the deep of the brain cerebral sulci and gyrus and controlled blood glucose levels were evaluated. The umbilical artery pulsation index(UAPI), middle cerebral artery pulsation index(MCAPI) and ductus venosus pulsation index(DVPI) among the 3 groups were compared, and the differences in fetal blood perfusion among the 3 groups were evaluated.Results:There were no significant differences in the depths of the sylvian fissure, parietooccipital sulci, calcarine sulci and cinguli sulci between the control group and the GDM group 1 (all P>0.05), and they were larger than those of the GDM group 2 (all P<0.05). The depths of lateral fissure, parieto-occipital sulcus, cingulate sulcus and calcarine sulcus were negatively correlated with fasting blood glucose, 1 h and 2 h postprandial blood glucose (all P<0.05). There were no significant differences in MCAPI, UAPI and DVPI between the control group and GDM1 group (all P>0.05). The MCAPI in GDM 2 group was lower than that in the control group and GDM 1 group, and the UAPI and DVPI values were higher than those in the control group and GDM1 group(all P<0.05). Conclusions:The maturity of fetal cerebral sulci and gyrus in GDM pregnant women is related to the blood glucose control of pregnant women. The change of blood perfusion caused by persistent hyperglycemia in pregnant women and intrauterine hypoxia may cause the development retardation of cerebral sulci and gyrus.
7.Mechanisms of Fufang Biejia Ruangan Pills Against Alcoholic Liver Disease via Regulating Liver-brain Dialogue Mediated by HMGB1-BDNF Axis
Yudong LIU ; Xiangying YAN ; Tao LI ; Chu ZHANG ; Bingbing CAI ; Zhaochen MA ; Na LIN ; Yanqiong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):214-223
ObjectiveTo systematically and objectively characterize the pharmacological effects of Fufang Biejia Ruangan pills (FBRP) in the intervention of alcoholic liver disease (ALD) using acute and chronic ALD mouse models and to elucidate its molecular mechanisms. MethodFifty SPF-grade male BALB/c mice were randomly divided into the normal group, model group, and FBRP low-, medium-, and high-dose groups (9.6, 19.2, 38.4 mg·kg-1). Except for the normal group, the remaining groups were given 56° white wine by gavage to establish the acute ALD model, with samples collected after 4 weeks. Thirty SPF-grade male C57BL/6N mice were randomly divided into the normal group, model group, and FBRP medium-dose group (19.2 mg·kg-1). The chronic ALD mouse model was established using the Lieber-DeCarli method over a 10-week period. Inflammatory markers in liver tissues were assessed using hematoxylin-eosin (HE), Sirius Red, oil red O staining, and enzyme-linked immunosorbent assay (ELISA). Intoxication behaviors of each group were objectively evaluated through sobering-up time, net-catching, and pole-climbing tests. Further bioinformatics analyses based on clinical transcriptomic data were conducted to identify key targets and molecular mechanisms of FBRP in alleviating ALD through liver-brain dialogue, with experimental validation by ELISA, Western blot, and immunohistochemical staining. ResultCompared with the normal group, the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in liver tissues of mice in the acute and chronic ALD model groups were significantly increased (P<0.05). Compared with the model group, the levels of AST and ALT in liver tissue of mice in FBRP groups were significantly decreased (P<0.05). Compared with the normal group, the time of grasping the net and climbing the pole in the acute ALD model group was significantly decreased within 4 weeks (P<0.01). Compared with the model group, the grasping and climbing time of FBRP high dose groups increased significantly within 4 weeks (P<0.05). Compared with the normal group, the expression of high mobility group protein B1 (HMGB1) protein in liver tissue and prefrontal lobe tissue of mice in the chronic ALD model group was significantly increased (P<0.01). Compared with the model group, the expression of HMGB1 protein in FBRP medium dose group was significantly decreased (P<0.05,P<0.01). Compared with the normal group, the expression of brain-derived neurotrophic factor (BDNF) protein and the release of γ-aminobutyric acid (GABA) in the prefrontal cortex of the model group were significantly decreased (P<0.01). Compared with the model group, the expression of BDNF protein and the release of GABA in the FBRP medium dose group were significantly increased (P<0.05). ConclusionThis study revealed that FBRP improved key pathological changes in ALD by modulating liver-brain dialogue mediated by the HMGB1-BDNF axis. These findings provide experimental evidence for the clinical use of FBRP in treating ALD and offer new insights for the development of ALD therapeutic agents.
8.Identification of in Vitro and in vivo Chemical Constituents of Ruyi Zhenbaowan Based on UHPLC-Q Exactive Orbitrap HRMS
Kedian CHEN ; Zhaochen MA ; Bingbing CAI ; Ying LIU ; Yudong LIU ; Tao LI ; Mingzhu XU ; Haiping WANG ; Na LIN ; Yanqiong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):78-84
ObjectiveTo identify the chemical constituents of Ruyi Zhenbaowan in vitro and in vivo. MethodThe chemical constituents of Ruyi Zhenbaowan were identified based on UHPLC-Q Exactive Orbitrap HRMS. A total of 12 male SD rats were randomized into two groups: control (pure water) and Ruyi Zhenbaowan (1.8 g·kg-1). The rats were administrated with the suspension of Ruyi Zhenbaowan or pure water by gavage. After 1.5 h, the plasma and cerebrospinal fluid were collected. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C18 column (2.1 mm × 150 mm, 1.7 μm) with a mixture of 0.1% formic acid aqueous solution (A) and acetonitrile (B) as the mobile phase. Gradient elution was carried out according to the procedure of 0~15 min,97%~80%A;15~30 min ,80%~60%A;30~40 min,60%~30%A;40~45 min,30%~5%A. The ion source was electrospray ionization, and scan range was m/z 100-1 500. The prototype components and the components in the plasma and cerebrospinal fluid were analyzed qualitatively by scanning in positive and negative ion modes and identified by comparison with the data in published literature and the information of standard substances. ResultA total of 126 chemical constituents were identified from the 80% methanol solution of Ruyi Zhenbaowan, and 14 and 7 prototype constituents were detected in the plasma and the cerebrospinal fluid, respectively. In addition, the fragmentation rules of apigenin, apigenin-7-O-glucuronide, galangin, liquiritin, piperine, glycyrrhizic acid, eugenol, gallic acid, and cholic acid were deduced. ConclusionThis study achieved rapid multicomponent characterization and identification of Ruyi Zhenbaowan in vitro and in vivo, providing theoretical support for exploring active substances and performing quality control.l.
9.Preliminary study of quantitative parameters from gastric tumor and spleen CT to predict the clinical stage of gastric cancer
Dongbo LYU ; Pan LIANG ; Mengru LIU ; Pengchao ZHAN ; Zhiwei HU ; Bingbing ZHU ; Songwei YUE ; Jianbo GAO
Chinese Journal of Radiology 2024;58(9):923-928
Objective:To investigate the value of CT quantitative parameters of tumor and spleen in predicting the clinical stage of gastric cancer (Ⅰ/Ⅱ stage and Ⅲ/Ⅳ stage).Methods:This study was a case-control study. The data of 145 patients with gastric cancer confirmed by pathology in the First Affiliated Hospital of Zhengzhou University from February 2019 to June 2021 were retrospectively collected, including 70 cases of Ⅰ/Ⅱ stage and 75 cases of Ⅲ/Ⅳ stage. On the baseline CT images, the tumor related parameters, including tumor thickness, length of tumor, CT attenuation of tumor unenhanced phase, CT attenuation of tumor arterial phase, CT attenuation of tumor venous phase were measured. The spleen related parameters, including splenic thickness, CT attenuation of splenic unenhanced phase, CT attenuation of splenic arterial phase, CT attenuation of splenic venous phase, and standard deviation of CT attenuation (CTsd) in splenic unenhanced phase were also measured. The independent sample t test or Mann-Whitney U test was used to compare the parameters between the Ⅰ/Ⅱ stage and Ⅲ/Ⅳ stage patients. The multi-factor logistic regression analysis was used to find the independent predictors of gastric cancer clinical stage, and establish the combined parameters. The efficiency to the diagnosis of gastric cancer stage of single and combined parameters was evaluated using the operating characteristic curve, and the DeLong test was used to compare the differences of area under the curve (AUC). Results:There were significant differences in tumor thickness, length of tumor, CT attenuation of tumor venous phase, CT attenuation of splenic unenhanced phase, CT attenuation of splenic venous phase, CTsd in splenic unenhanced phase between the Ⅰ/Ⅱ stage and Ⅲ/Ⅳ stage of gastric cancer ( P<0.05). Multivariate analysis showed that tumor thickness ( OR=1.073, 95% CI 1.026-1.123, P=0.002), CT attenuation of splenic venous phase ( OR=1.040, 95% CI 1.011-1.070, P=0.006) and CTsd in splenic unenhanced phase ( OR=1.625, 95% CI 1.330-1.987, P<0.001) were independent risk factors for the clinical stage of gastric cancer and the combined parameters were established. The AUC values of tumor thickness, CT attenuation of splenic venous phase, CTsd in splenic unenhanced phase and combined parameters were 0.655, 0.614, 0.749 and 0.806, respectively. The AUC of combined parameters was higher than those of tumor thickness and CT attenuation of splenic venous phase, and the differences were statistically significant ( Z=3.37, 3.82, both P<0.001). Conclusion:Tumor thickness, CT attenuation of splenic venous phase and CTsd in splenic unenhanced phase are independent risk factors for the clinical stage of gastric cancer, and combined parameters can improve the diagnostic efficiency.
10.Improvement effect of down-regulation of miR-208a on 5-FU resistance in colorectal cancer cells through targeting SFRP1 for mediating Wnt signaling pathway
Bingbing HU ; Kangning LUO ; Su PENG ; Yuzhong ZHOU ; Maoliang CHEN ; Changhua LIU
Journal of Jilin University(Medicine Edition) 2024;50(4):947-955
Objective:To discuss the effect of downregulating microRNA-208a(miR-208a)on the resistance of the colorectal cancer cells to 5-fluorouracil(5-FU),and to clarify its related molecular mechanism.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-208a and secreted frizzled-related protein 1(SFRP1)mRNA in the 5-FU-resistant colorectal cancer cell line HT-29/5-FU and its parent HT-29 cells.The HT-29/5-FU cells were transfected with miR-208a inhibitor plasmid and its negative control plasmid(inhibitor-NC),and SFRP1 small interfering RNA(si-SFRP1)and its negative control plasmid(si-NC),either separately or in combination,followed by treatment with 5-FU.The cells were divided into inhibitor-NC group,miR-208a inhibitor group,miR-208a inhibitor+si-NC group,and miR-208a inhibitor+si-SFRP1 group.MTT assay was used to detect the proliferation activities of the cells and the resistance indexes were calculated;Annexin V-FITC/PI double staining and flow cytometry were used to detect the apoptotic rates of the cells after treated with different concentrations of 5-FU;Western blotting method was used to detect the expression levels of SFRP1,β-catenin,P-glycoprotein(P-gp),and ATP-binding cassette subfamily B member 1(ABCB1)proteins in the cells in various groups;dual-luciferase reporter gene assay was used to validate the targeting relationship between miR-208a and SFRP1.Results:Compared with HT-29 cells,the expression level of miR-208a in the HT-29/5-FU cells was increased(P<0.05),and the expression level of SFRP1 mRNA was decreased(P<0.05).Compared with inhibitor-NC group,the proliferation activity of the cells in miR-208a inhibitor group was decreased(P<0.05),the resistance index was decreased,the apoptotic rate was increased(P<0.05),and the expression levels of β-catenin,P-gp,and ABCB1 proteins in the cells were decreased(P<0.05).The dual-luciferase reporter gene assay results showed that SFRP1 was a target gene of miR-208a and miR-208a could negatively regulate the expression of SFRP1.Compared with miR-208a inhibitor+si-NC group,the proliferation activity of the cells in miR-208a inhibitor+si-SFRP1 group was increased(P<0.05),the resistance index was increased,the apoptotic rate was decreased(P<0.05),and the expression levels of β-catenin,P-gp,and ABCB1 proteins in the cells were increased(P<0.05).Conclusion:Downregulation of miR-208a can improve the resistance of the HT-29/5-FU cells to 5-FU by targeting and upregulating the SFRP1 expression,thereby inhibiting the transmission of the Wnt signaling pathway.

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