1.Scutellarin exerts protective effects against atherosclerosis by regulating PERK-Nrf2/ATF4-CHOP signaling pathways
Shi-qi SHAN ; Rui-qi ZHAO ; Yue JIN
Chinese Pharmacological Bulletin 2025;41(4):738-745
Aim To investigate the effect of synthetic small interfering RNA(siRNA)targetingβ-arrestin2 on the apoptosis of hepatic stellate cells(HSC)in vitro.Methods The protective effect of scutellarin on high-fat diet and vitamin D3-induced AS rats.Forty male SD rats weighing 180~220 g were randomly divided into the normal group,the high-fat diet group and the low and high-dose scutellarin.A week of adaptive feeding later,the normal group ate normal diet,the oth-er groups ate a high-fat diet and intraperitoneal injec-tion of vitamin D3,while the administration group was intraperitoneally injected with different doses of scutel-larin daily.The animals were anesthetized after 12 weeks,and the rat aortic blood and aortic blood vessels were taken out.Rat aortic blood vessels were stained with hematoxylin-eosin(HE)to observe the pathologi-cal changes.Tissue superoxide dismutase(SOD),ma-londialdehyde(MDA),glutathione peroxidase(GSH-Px)and glutathione-sulfase(GSH-ST)and total cho-lesterol(TC)),the content of triglyceride(TG),low density lipoprotein(LDL-C),high density lipoprotein(HDL-C)were detected.The mRNA expression of PERK and eIF2α in tissues was detected by qRT-PCR.The expressions of GRP78,p-PERK,PERK,p-eIF2α,eIF2α,ATF4,CHOP,Nrf2,Bcl-2,PUMA,caspase-3 and caspase-12 were detected by Western blot.Re-sults The results of HE staining showed that the foam cells and lipid deposition under the intima of the scute-llarin group were reduced,which proved that scutellarin had a certain protective effect on vascular endothelial cells.In vivo the expression of TC,TG and LDL-C in scutellarin group decreased and the expression of HDL-C increased.Scutellarin could down-regulate the pro-tein expression of PERK and eIF2α,and down-regulate the expression of GRP78,p-PERK,p-eIF2α,ATF4,CHOP,PUMA,caspase-3,caspase-12 and up-regulated Nrf2 and Bcl-2.The anti-AS effect of scutellarin was proved to be closely related to its regulation of PERK-Nrf2/ATF4-CHOP signaling pathway.Conclusion Scutellarin inhibits endoplasmic reticulum stress and apoptosis by regulating the PERK-Nrf2/ATF4-CHOP signaling pathway,thereby treating AS.
2.Established cell model and mechanism of visceral hypersensitivity and nerve hyperplasia in IBS using P815 and N2a co-culture
Hongbin LI ; Chunli GAN ; Xiangyu XIE ; Shan LIU ; Qin LU ; Wei KE ; Shi-yu QI ; Yusheng HUANG ; Hongmei TANG
Chinese Journal of Pathophysiology 2025;41(4):825-832
AIM:To establish a cell model of visceral hypersensitivity and nerve hyperplasia in irritable bowel syndrome(IBS)by conducting an in vitro co-culture of mouse P815 mast cells and N2a nerve cells and explore its possible mechanism.METHODS:Enzyme-linked immunosorbent assay(ELISA)with three replicates was used to confirm the C48/80-induced P815 degranulation.The length of neurites was observed under bright field microscopy to determine the number of differentiated neurons,thereby selecting the concentration of retinoic acid(RA)for stimulating the differentia-tion of N2a cells,with four replicates.A co-culture system of P815 and N2a cells was established using Transwell cham-bers with four replicates.The following groups were established:N2a cells cultured alone,N2a cells co-cultured with P815 cells,N2a cells co-cultured with P815 cells plus C48/80,and N2a cells plus RA group.After co-culturing,the num-ber of differentiated N2a cells was observed under bright field.The expression of nerve growth factor(NGF),tyrosine ki-nase receptor A(TRKA),growth-associated protein-43(GAP43),neuron-specific enolase(NSE),synapsin(SYN),and postsynaptic density protein-95(PSD-95)at both protein and gene levels in N2a cells was detected using Western blot and polymerase chain reaction(PCR),with four replicates.RESULTS:The best condition for N2a differentiation was stimulation with 10 μmol/L RA for 24 hours,whereas the best condition for degranulation was stimulation of P815 cells with 20 mg/L C48/80 for 24 hours.Compared with N2a cells cultured alone,the differentiation ratio of the N2a+P815+C48/80 and N2a+RA groups was significantly increased(P<0.01),and the protein and mRNA expressions of NGF,TRKA,GAP43,NSE,SYN,and PSD-95 were significantly increased(P<0.05).CONCLUSION:Our results revealed that mast cell degranulation enhances the level of nerve hyperplasia in enteric nerve cells and promotes changes in nerve structure and function.Synaptic remodeling regulated by abnormal expression of key proteins such as NGF,TRKA,and GAP43 is involved in the nerve hyperplasia induced by mast cell degranulation.
3.Scutellarin exerts protective effects against atherosclerosis by regulating PERK-Nrf2/ATF4-CHOP signaling pathways
Shi-qi SHAN ; Rui-qi ZHAO ; Yue JIN
Chinese Pharmacological Bulletin 2025;41(4):738-745
Aim To investigate the effect of synthetic small interfering RNA(siRNA)targetingβ-arrestin2 on the apoptosis of hepatic stellate cells(HSC)in vitro.Methods The protective effect of scutellarin on high-fat diet and vitamin D3-induced AS rats.Forty male SD rats weighing 180~220 g were randomly divided into the normal group,the high-fat diet group and the low and high-dose scutellarin.A week of adaptive feeding later,the normal group ate normal diet,the oth-er groups ate a high-fat diet and intraperitoneal injec-tion of vitamin D3,while the administration group was intraperitoneally injected with different doses of scutel-larin daily.The animals were anesthetized after 12 weeks,and the rat aortic blood and aortic blood vessels were taken out.Rat aortic blood vessels were stained with hematoxylin-eosin(HE)to observe the pathologi-cal changes.Tissue superoxide dismutase(SOD),ma-londialdehyde(MDA),glutathione peroxidase(GSH-Px)and glutathione-sulfase(GSH-ST)and total cho-lesterol(TC)),the content of triglyceride(TG),low density lipoprotein(LDL-C),high density lipoprotein(HDL-C)were detected.The mRNA expression of PERK and eIF2α in tissues was detected by qRT-PCR.The expressions of GRP78,p-PERK,PERK,p-eIF2α,eIF2α,ATF4,CHOP,Nrf2,Bcl-2,PUMA,caspase-3 and caspase-12 were detected by Western blot.Re-sults The results of HE staining showed that the foam cells and lipid deposition under the intima of the scute-llarin group were reduced,which proved that scutellarin had a certain protective effect on vascular endothelial cells.In vivo the expression of TC,TG and LDL-C in scutellarin group decreased and the expression of HDL-C increased.Scutellarin could down-regulate the pro-tein expression of PERK and eIF2α,and down-regulate the expression of GRP78,p-PERK,p-eIF2α,ATF4,CHOP,PUMA,caspase-3,caspase-12 and up-regulated Nrf2 and Bcl-2.The anti-AS effect of scutellarin was proved to be closely related to its regulation of PERK-Nrf2/ATF4-CHOP signaling pathway.Conclusion Scutellarin inhibits endoplasmic reticulum stress and apoptosis by regulating the PERK-Nrf2/ATF4-CHOP signaling pathway,thereby treating AS.
4.A near-complete genomic analysis of aggregated outbreaks of norovirus subtype GⅡ.17P17 in Beijing Chaoyang District from 2014 to 2024
Xiangyu HU ; Jianhong ZHAO ; Shan WANG ; Xiao QI ; Taoli HAN ; Yanhui YANG ; Yan GAO ; Shi CONG ; Lijiao CAO ; Lingli SUN ; Miao JIN ; Yang JIAO
Chinese Journal of Preventive Medicine 2025;59(5):640-649
Objective:To examine the near-complete genomic analysis of norovirus (NoV) subtype GⅡ.17 [P17] outbreaks in Beijing Chaoyang District from 2014 to 2024.Methods:Data and specimens related to outbreaks of the NoV aggregation in Beijing′s Chaoyang District from 2014 to 2024 were collected. The NoV was identified using real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR). Specimens with positive nucleic acid were amplified by standard PCR, whole genome sequencing and evolutionary analysis. Amino acid site variations were compared.Results:In Chaoyang District, from 2014 to 2024, a total of 637 aggregated outbreaks caused by the NoV infection were reported, of which 584 were successfully typed. The epidemic caused by the GⅡ.17 [P17] subtype accounted for 8.79% (56/637), which was the dominant epidemic gene subtype in 2014-2015, sporadic in 2016-2019, reappeared in 2022, and significantly increased in 2024 (27.27%, 24/88). Outbreaks caused by the GⅡ.17 [P17] subtype occurred mainly from October to December, with the main sites of occurrence in primary schools and kindergartens. This study yielded 53 near-complete genome sequences of the GⅡ.17 [P17] subtype from 46 incidents in Chaoyang District. The GⅡ.17 [P17] subtype sequences of Chaoyang District from 2014 to 2024 were segmented into three subgroups on the evolutionary tree, with sequences from 2014 to 2019, 2022 to April 2024, and May to December 2024 clustered into the d, e, and b subgroups, respectively. In the VP1 region′s P2 area, particularly at the HBGA binding site, subgroups b and e exhibited mutations in 22 and two sites, while subgroups b and e showed mutations in four and one sites, predominantly in the RdRp region.Conclusion:The outbreak caused by the NoV GⅡ.17 [P17] subtype in Chaoyang District from 2014 to 2024 continues, with a significant increase in 2024, and it becomes the dominant gene subtype from October to December. The sequence formation of the NoV GⅡ.17 [P17] subtype in Chaoyang District from January to April 2022 and from May to December 2024 shows two different evolutions, with specific mutation sites, requiring continuous monitoring of the NoV GⅡ.17 [P17] subtype.
5.Development of intelligent equipment for rapid microbial detection of Atractylodis Macrocephalae Rhizoma decoction pieces based on measurement technology for traditional Chinese medicine manufacturing.
Yang LIU ; Wu-Zhen QI ; Yu-Tong WU ; Shan-Xi ZHU ; Xiao-Jun ZHAO ; Qia-Tong XIE ; Yu-Feng GUO ; Jing ZHAO ; Nan LI ; Shi-Jun WANG ; Qi-Hui SUN ; Zhi-Sheng WU
China Journal of Chinese Materia Medica 2025;50(16):4610-4618
Microbial detection and control of traditional Chinese medicine(TCM) decoction pieces are crucial for the quality control of TCM preparations. It is also a key area of research in the measurement technology and equipment development for TCM manufacturing. Guided by TCM manufacturing measurement methodologies, this study presented a design of a novel portable microbial detection device, using Atractylodis Macrocephalae Rhizoma decoction pieces as a demonstration. Immunomagnetic separation technology was employed for specific isolation and labeling of target microorganisms. Enzymatic signal amplification was utilized to convert weak biological signals into colorimetric signals, constructing an optical biosensor. A self-developed smartphone APP was further applied to analyze the colorimetric signals and quantify target concentrations. A portable and automated detection system based on Arduino microcontroller was developed to automatically perform target microbial separation/extraction, as well as mimetic enzyme labeling and catalytic reactions. The developed equipment specifically focuses on the rapid and quantitative microbial analysis of TCM active pharmaceutical ingredients, intermediates in TCM manufacturing, and final TCM products. Experimental results demonstrate that the equipment could detect Salmonella in samples within 2 h, with a detection limit as low as 5.1 × 10~3 CFU·mL~(-1). The equipment enables the rapid detection of microorganisms in TCM decoction pieces, providing a potential technical solution for on-site rapid screening of microbial contamination indicators in TCM. It has broad application prospects in measurement technology for TCM manufacturing and offers strong technical support for the modernization, industrialization, and intelligent development of TCM.
Drugs, Chinese Herbal/analysis*
;
Atractylodes/microbiology*
;
Rhizome/microbiology*
;
Biosensing Techniques/methods*
;
Medicine, Chinese Traditional
;
Colorimetry/instrumentation*
;
Quality Control
6.Machine learning-assisted microfluidic approach for broad-spectrum liposome size control.
Yujie JIA ; Xiao LIANG ; Li ZHANG ; Jun ZHANG ; Hajra ZAFAR ; Shan HUANG ; Yi SHI ; Jian CHEN ; Qi SHEN
Journal of Pharmaceutical Analysis 2025;15(6):101221-101221
Liposomes serve as critical carriers for drugs and vaccines, with their biological effects influenced by their size. The microfluidic method, renowned for its precise control, reproducibility, and scalability, has been widely employed for liposome preparation. Although some studies have explored factors affecting liposomal size in microfluidic processes, most focus on small-sized liposomes, predominantly through experimental data analysis. However, the production of larger liposomes, which are equally significant, remains underexplored. In this work, we thoroughly investigate multiple variables influencing liposome size during microfluidic preparation and develop a machine learning (ML) model capable of accurately predicting liposomal size. Experimental validation was conducted using a staggered herringbone micromixer (SHM) chip. Our findings reveal that most investigated variables significantly influence liposomal size, often interrelating in complex ways. We evaluated the predictive performance of several widely-used ML algorithms, including ensemble methods, through cross-validation (CV) for both liposome size and polydispersity index (PDI). A standalone dataset was experimentally validated to assess the accuracy of the ML predictions, with results indicating that ensemble algorithms provided the most reliable predictions. Specifically, gradient boosting was selected for size prediction, while random forest was employed for PDI prediction. We successfully produced uniform large (600 nm) and small (100 nm) liposomes using the optimised experimental conditions derived from the ML models. In conclusion, this study presents a robust methodology that enables precise control over liposome size distribution, offering valuable insights for medicinal research applications.
8.Expert consensus on evaluation index system construction for new traditional Chinese medicine(TCM) from TCM clinical practice in medical institutions.
Li LIU ; Lei ZHANG ; Wei-An YUAN ; Zhong-Qi YANG ; Jun-Hua ZHANG ; Bao-He WANG ; Si-Yuan HU ; Zu-Guang YE ; Ling HAN ; Yue-Hua ZHOU ; Zi-Feng YANG ; Rui GAO ; Ming YANG ; Ting WANG ; Jie-Lai XIA ; Shi-Shan YU ; Xiao-Hui FAN ; Hua HUA ; Jia HE ; Yin LU ; Zhong WANG ; Jin-Hui DOU ; Geng LI ; Yu DONG ; Hao YU ; Li-Ping QU ; Jian-Yuan TANG
China Journal of Chinese Materia Medica 2025;50(12):3474-3482
Medical institutions, with their clinical practice foundation and abundant human use experience data, have become important carriers for the inheritance and innovation of traditional Chinese medicine(TCM) and the "cradles" of the preparation of new TCM. To effectively promote the transformation of new TCM originating from the TCM clinical practice in medical institutions and establish an effective evaluation index system for the transformation of new TCM conforming to the characteristics of TCM, consensus experts adopted the literature research, questionnaire survey, Delphi method, etc. By focusing on the policy and technical evaluation of new TCM originating from the TCM clinical practice in medical institutions, a comprehensive evaluation from the dimensions of drug safety, efficacy, feasibility, and characteristic advantages was conducted, thus forming a comprehensive evaluation system with four primary indicators and 37 secondary indicators. The expert consensus reached aims to encourage medical institutions at all levels to continuously improve the high-quality research and development and transformation of new TCM originating from the TCM clinical practice in medical institutions and targeted at clinical needs, so as to provide a decision-making basis for the preparation, selection, cultivation, and transformation of new TCM for medical institutions, improve the development efficiency of new TCM, and precisely respond to the public medication needs.
Medicine, Chinese Traditional/standards*
;
Humans
;
Consensus
;
Drugs, Chinese Herbal/therapeutic use*
;
Surveys and Questionnaires
9.A near-complete genomic analysis of aggregated outbreaks of norovirus subtype GⅡ.17P17 in Beijing Chaoyang District from 2014 to 2024
Xiangyu HU ; Jianhong ZHAO ; Shan WANG ; Xiao QI ; Taoli HAN ; Yanhui YANG ; Yan GAO ; Shi CONG ; Lijiao CAO ; Lingli SUN ; Miao JIN ; Yang JIAO
Chinese Journal of Preventive Medicine 2025;59(5):640-649
Objective:To examine the near-complete genomic analysis of norovirus (NoV) subtype GⅡ.17 [P17] outbreaks in Beijing Chaoyang District from 2014 to 2024.Methods:Data and specimens related to outbreaks of the NoV aggregation in Beijing′s Chaoyang District from 2014 to 2024 were collected. The NoV was identified using real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR). Specimens with positive nucleic acid were amplified by standard PCR, whole genome sequencing and evolutionary analysis. Amino acid site variations were compared.Results:In Chaoyang District, from 2014 to 2024, a total of 637 aggregated outbreaks caused by the NoV infection were reported, of which 584 were successfully typed. The epidemic caused by the GⅡ.17 [P17] subtype accounted for 8.79% (56/637), which was the dominant epidemic gene subtype in 2014-2015, sporadic in 2016-2019, reappeared in 2022, and significantly increased in 2024 (27.27%, 24/88). Outbreaks caused by the GⅡ.17 [P17] subtype occurred mainly from October to December, with the main sites of occurrence in primary schools and kindergartens. This study yielded 53 near-complete genome sequences of the GⅡ.17 [P17] subtype from 46 incidents in Chaoyang District. The GⅡ.17 [P17] subtype sequences of Chaoyang District from 2014 to 2024 were segmented into three subgroups on the evolutionary tree, with sequences from 2014 to 2019, 2022 to April 2024, and May to December 2024 clustered into the d, e, and b subgroups, respectively. In the VP1 region′s P2 area, particularly at the HBGA binding site, subgroups b and e exhibited mutations in 22 and two sites, while subgroups b and e showed mutations in four and one sites, predominantly in the RdRp region.Conclusion:The outbreak caused by the NoV GⅡ.17 [P17] subtype in Chaoyang District from 2014 to 2024 continues, with a significant increase in 2024, and it becomes the dominant gene subtype from October to December. The sequence formation of the NoV GⅡ.17 [P17] subtype in Chaoyang District from January to April 2022 and from May to December 2024 shows two different evolutions, with specific mutation sites, requiring continuous monitoring of the NoV GⅡ.17 [P17] subtype.
10.Preliminary study on the construction of an echocardiogram image quality control system based on artificial intelligence
Zhanru QI ; Hanlin CHENG ; Chunjie SHAN ; Ruiyang CHEN ; Hexiang WENG ; Yue DU ; Guanjun GUO ; Xiaoxian WANG ; Jing YAO ; Shouhua LUO ; Aijuan FANG ; Hui CHEN ; Zhongqing SHI
Chinese Journal of Ultrasonography 2025;34(2):107-113
Object:To explore the feasibility of using artificial intelligence for quality control of echocardiographic images.Methods:Retrospectively,5 000 two-dimensional echocardiographic video images within the period from 2021 to 2023 were randomly retrieved from the echocardiography database of Nanjing Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University. Among these selected images,1 559 of them were apical views. The physician team formulated the scoring rules,which specifically included four scoring criteria:gain,scaling ratio,cardiac axis angle,and structure. Subsequently,the data were labeled with view classification and image quality scores. The labeled data were further partitioned into the training set( n = 643),the validation set( n = 276),and the test set( n = 640). The training and validation sets were utilized for constructing the models for view classification and quality assessment,while the test set was employed to verify the models' effectiveness. The view classification module was implemented using the SlowFast model,and the quality assessment module involved algorithms such as ResNet,Video Swin Transformer,SSD,and U-Net. Results:The average accuracy,precision,recall rate and F1 score of the classification model in identifying each apical view were 0.987 1,0.983 0,0.987 1 and 0.984 9 respectively,and the inference time was(333.4 ± 105.4)ms. The average accuracies of the quality assessment module in terms of gain,scaling ratio,cardiac axis angle and display of main structures were 0.915 1,0.928 2,0.938 7 and 0.965 6 respectively,and the overall scoring accuracy was 0.912 7.Conclusions:The echocardiogram quality control system developed in this research can effectively classify and evaluate the quality of two-dimensional images of the apical views in echocardiograms. Moreover,it guarantees the objectivity,timeliness and high-efficiency of quality control,which has reference value for the establishment of the echocardiogram quality control system.

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