1.Study on the Anti-Atherosclerotic Mechanism of Tiaozhi Xiaoban Mixture
Meng LIU ; Danning ZHANG ; Junnan ZENG ; Lei LU ; Tian LIANG ; Ying XU ; Tong CHEN ; Xin ZHAO ; Hanmei ZHANG ; Yong BIAN ; Zhongliang WANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(9):1178-1188
OBJECTIVE To explore the ameliorative effect of Tiaozhi Xiaoban Mixture on atherosclerosis and the potential role of long non-coding RNA(Linc RNA)in anti-atherosclerosis.METHODS A model of atherosclerosis was established in SD rats subjec-ted to a high-fat diet.At 4 weeks post-modeling,thoracic aortic tissues from atherosclerotic rats were collected for hematoxylin-eosin(HE)staining to systematically evaluate the anti-atherosclerotic effects of Tiaozhi Xiaoban Mixture at different doses.Biochemical kits were utilized to assess relevant indices related to blood lipid levels as well as liver and kidney function,thereby evaluating the impact of Tiaozhi Xiaoban Mixture on these parameters.Enzyme-linked immunosorbent assay(ELISA)was employed to measure serum inflam-mation markers influenced by Tiaozhi Xiaoban Mixture.Additionally,TUNEL staining and Western blot analysis were conducted to ex-amine the apoptotic effects of Tiaozhi Xiaoban Mixture on thoracic aorta tissue.Finally,qPCR was used to detect the expression levels of Line-HC,MALAT1,etc.,in order to evaluate how Tiaozhi Xiaoban Mixture affecting these specific RNA molecules.RESULTS Following treatment with Tiaozhi Xiaoban Mixture,the blood lipid profiles indicated that total cholesterol(TC),triglycerides(TG),and low-density lipoprotein cholesterol(LDL-C)were significantly down-regulated(P<0.05,P<0.01),while high-density lipopro-tein cholesterol(HDL-C)levels were up-regulated in the atherosclerotic rats.Moreover,serum levels of liver and kidney function markers such as aspartate aminotransferase(AST),alanine aminotransferase(ALT),blood urea nitrogen(BUN),and creatinine(Cr)exhibited down-regulation(P<0.05,P<0.01).Additionally,pro-inflammatory factors including interleukin-6(IL-6),interleukin-1 beta(IL-1β),tumor necrosis factor-alpha(TNF-α),high-sensitivity C-reactive protein(hs-CRP),and matrix metallopeptidase 9(MMP-9)were also reduced(P<0.01),whereas the anti-inflammatory factor interleukin-10(IL-10)was found to be elevated(P<0.01).Furthermore,after oral administration of Tiaozhi Xiaoban Mixture,expression levels of apoptosis-related factors NLRP3,ASC,Cleaved Caspase-1,Cleaved IL-1 β,Puma,Bax,Noxa,and MDM2 in thoracic aorta tissues from the atherosclerotic rats showed sig-nificant down-regulation(P<0.05,P<0.01).Notably,following treatment with Tiaozhi Xiaoban Mixture,mRNA levels of Linc-HC decreased while mRNA expression of MALAT1 increased(P<0.05,P<0.01).CONCLUSION Tiaozhi Xiaoban Mixture may inhibit the expression of Linc-HC and up-regulate the expression of MALAT1 to reduce the formation of atherosclerotic plaque,improve ab-normal blood lipids and liver and kidney function,alleviate inflammation and inhibit apoptosis.
2.Anti-inflammatory and anti-apoptotic effects and mechanism of total flavonoids of hawthorn leaves on rat intestinal epithelial cells
Kai WANG ; Pei LIU ; Kexin QI ; Jingyi WANG ; Chenlu SUN ; Danning SHI ; Hongyue CHEN ; Daoling HE ; Yan ZHU ; Ling GAN
Chinese Journal of Veterinary Science 2025;45(7):1450-1457
This study aims to investigate the anti-inflammatory and anti-apoptotic effects of total flavonoids of hawthorn leaves(TFHL)on lipopolysaccharide(LPS)-induced inflammatory injury in rat intestinal epithelial(IEC-6)cells,as well as the underlying mechanisms.An in vitro inflam-mation model was first established by treating IEC-6 cells with lipopolysaccharide(LPS).IEC-6 cells were then incubated with three concentrations of TFHL for 24 h prior to a further 24 h LPS treatment.RT-qPCR was used to quantify mRNA levels of the inflammatory genes COX-2 and iN-OS,while Western blotting was used to assess protein levels of the apoptotic markers Bax,cleaved Caspase-3,Bcl-2,and the JNK/p-JNK signaling pathway.Finally,cells were pretreated with TFHL and/or the JNK inhibitor SP600125 for 24 h before LPS exposure for 24 h,in order to evaluate the combined effects of TFHL and SP600125 on LPS-induced inflammatory cytokine expression and apoptotic protein levels in IEC-6 cells.The results showed that,compared with the LPS group,the mRNA level of COX-2 and iNOS in the 2.5,5.0,10.0 mg/L TFHL group and the Bax and Caspase-3 protein levels decreased significantly(P<0.01),and the Bcl-2 protein level was significantly higher(P<0.01),p-JNK protein level and p-JNK/JNK ratio decreased significantly(P<0.01);compared with the LPS group,the COX-2 and iNOS mRNA levels of the TFHL+LPS group de-creased significantly(P<0.01),Bax,and Caspase-3 protein levels decreased significantly(P<0.01),and the level of Bcl-2 protein increased significantly(P<0.05);compared with the LPS group,the COX-2 and iNOS mRNA levels of the TFHL+SP600125 group decreased significantly(P<0.01),Bax and Caspase-3 protein levels decreased significantly(P<0.01),and Bcl-2 protein level increased significantly(P<0.01).These findings indicate that TFHL exerts anti-inflammato-ry and anti-apoptotic effects in LPS-challenged IEC-6 cells by inhibiting the JNK signaling path-way.
3.Study on the Anti-Atherosclerotic Mechanism of Tiaozhi Xiaoban Mixture
Meng LIU ; Danning ZHANG ; Junnan ZENG ; Lei LU ; Tian LIANG ; Ying XU ; Tong CHEN ; Xin ZHAO ; Hanmei ZHANG ; Yong BIAN ; Zhongliang WANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(9):1178-1188
OBJECTIVE To explore the ameliorative effect of Tiaozhi Xiaoban Mixture on atherosclerosis and the potential role of long non-coding RNA(Linc RNA)in anti-atherosclerosis.METHODS A model of atherosclerosis was established in SD rats subjec-ted to a high-fat diet.At 4 weeks post-modeling,thoracic aortic tissues from atherosclerotic rats were collected for hematoxylin-eosin(HE)staining to systematically evaluate the anti-atherosclerotic effects of Tiaozhi Xiaoban Mixture at different doses.Biochemical kits were utilized to assess relevant indices related to blood lipid levels as well as liver and kidney function,thereby evaluating the impact of Tiaozhi Xiaoban Mixture on these parameters.Enzyme-linked immunosorbent assay(ELISA)was employed to measure serum inflam-mation markers influenced by Tiaozhi Xiaoban Mixture.Additionally,TUNEL staining and Western blot analysis were conducted to ex-amine the apoptotic effects of Tiaozhi Xiaoban Mixture on thoracic aorta tissue.Finally,qPCR was used to detect the expression levels of Line-HC,MALAT1,etc.,in order to evaluate how Tiaozhi Xiaoban Mixture affecting these specific RNA molecules.RESULTS Following treatment with Tiaozhi Xiaoban Mixture,the blood lipid profiles indicated that total cholesterol(TC),triglycerides(TG),and low-density lipoprotein cholesterol(LDL-C)were significantly down-regulated(P<0.05,P<0.01),while high-density lipopro-tein cholesterol(HDL-C)levels were up-regulated in the atherosclerotic rats.Moreover,serum levels of liver and kidney function markers such as aspartate aminotransferase(AST),alanine aminotransferase(ALT),blood urea nitrogen(BUN),and creatinine(Cr)exhibited down-regulation(P<0.05,P<0.01).Additionally,pro-inflammatory factors including interleukin-6(IL-6),interleukin-1 beta(IL-1β),tumor necrosis factor-alpha(TNF-α),high-sensitivity C-reactive protein(hs-CRP),and matrix metallopeptidase 9(MMP-9)were also reduced(P<0.01),whereas the anti-inflammatory factor interleukin-10(IL-10)was found to be elevated(P<0.01).Furthermore,after oral administration of Tiaozhi Xiaoban Mixture,expression levels of apoptosis-related factors NLRP3,ASC,Cleaved Caspase-1,Cleaved IL-1 β,Puma,Bax,Noxa,and MDM2 in thoracic aorta tissues from the atherosclerotic rats showed sig-nificant down-regulation(P<0.05,P<0.01).Notably,following treatment with Tiaozhi Xiaoban Mixture,mRNA levels of Linc-HC decreased while mRNA expression of MALAT1 increased(P<0.05,P<0.01).CONCLUSION Tiaozhi Xiaoban Mixture may inhibit the expression of Linc-HC and up-regulate the expression of MALAT1 to reduce the formation of atherosclerotic plaque,improve ab-normal blood lipids and liver and kidney function,alleviate inflammation and inhibit apoptosis.
4.Construction Process and Quality Control Points of the Database for Facial Phenotypes and Clinical Data of Pediatric Growth and Development-related Diseases
Jiaqi QIANG ; Yingjing WANG ; Danning WU ; Runzhu LIU ; Jiuzuo HUANG ; Hui PAN ; Xiao LONG ; Shi CHEN
Medical Journal of Peking Union Medical College Hospital 2025;16(3):552-557
The growth and development of children is an important stage for health, and its monitoringand intervention are related to the long-term development of individuals. The construction of a standardized and multi-dimensional database of pediatric growth and development-related diseases is an important basis for realizing precise diagnosis and treatment and health management. Based on the needs of clinical practice, this study proposes to establish a specialized database of pediatric growth and development-related diseases that integrates facial phenotypes and clinical diagnosis and treatment information. This study elaborates on the construction process, including data sources, data collection content, and the operation and management of the database; and proposes key points for quality control, including the establishment of quality control nodes, database construction standards, and a full-process quality control framework. The above ensure the integrity, logic and effectiveness of the data, so that the database can provide an objective basis for the screening and diagnosis of pediatric growth and development-related diseases. On the basis of scientific data management and strict quality control, the database will help reveal the patterns of children's growth and development, and promote the level of children's health management.
5.Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein
Shuxian SHAO ; Yanmei WANG ; Yihan XU ; Jiaxin ZHENG ; Yufan ZHANG ; Danning LIU ; Yuan LI
Journal of Army Medical University 2025;47(11):1199-1207
Objective To develop an endothelialized microfluidic chip model that simulates the spatial architecture and bioactivity of retinal vasculature,enabling thrombosis modeling and thrombolytic efficacy validation.Methods A tri-level microvascular network chip(300/200/100 μm diameters)with bifurcated architecture was fabricated using soft lithography.Human retinal microvascular endothelial cells(HRMECs)were perfused into channels,with endothelial coverage monitored via phase-contrast microscopy and F-actin staining.Cellular bioactivity was assessed using mitochondrial membrane potential probes(5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide,JC-1)and nitric oxide(NO)quantification.Fresh blood samples from 10 healthy donors(Yongchuan Hospital Affiliated to Chongqing Medical University,March to June 2024)were perfused with digital injection pump to mimic blood flow in human body into 3 experimental groups:normal whole blood,and TNF-α-activated endothelium+normal blood,TNF-α-activated endothelium+TNF-α-treated blood.Three inlet blood flow rates of 37.8、11.1 and 3.5 μL/min were set in each group.Two experimental groups,normal saline and recombinant human tissue-type plasminogen activator(rtPA),were established using the endothelialized microfluidic thrombosis model to validate thrombolytic efficacy.Endothelial functional impacts were assessed through integrated DAPI/NO staining and thrombosis model analysis across 3 intervention phases:pre-thrombosis,post-thrombosis,and post-thrombolysis.Results A tri-level microfluidic vascular model(300/200/100 μm diameters)was successfully constructed.In 72 h after endothelial cell perfusion,complete channel coverage was achieved,with phase-contrast microscopy and F-actin staining confirming confluent cellular alignment.JC-1/NO assays validated preserved endothelial bioactivity.Compared with the whole blood group,both TNF-α-activated endothelium+normal blood and TNF-α-activated endothelium+TNF-α-treated blood groups exhibited significantly increased thrombus occupancy rates at identical flow rates(all P<0.001).Notably,TNF-α-activated endothelium+TNF-α-treated blood group demonstrated the highest thrombus ratio at 3.5 μL/min(P<0.001).The rtPA group showed superior thrombolytic efficacy versus saline(P<0.001).Endothelial monolayer integrity was maintained across intervention phases,with thrombosis triggering significant NO elevation(P<0.001).Conclusion Our retinal vasculature-mimetic microfluidic model enables precise thrombosis modeling and drug evaluation,providing new methodology for studying retinal vascular occlusive diseases.
6.Anti-inflammatory and anti-apoptotic effects and mechanism of total flavonoids of hawthorn leaves on rat intestinal epithelial cells
Kai WANG ; Pei LIU ; Kexin QI ; Jingyi WANG ; Chenlu SUN ; Danning SHI ; Hongyue CHEN ; Daoling HE ; Yan ZHU ; Ling GAN
Chinese Journal of Veterinary Science 2025;45(7):1450-1457
This study aims to investigate the anti-inflammatory and anti-apoptotic effects of total flavonoids of hawthorn leaves(TFHL)on lipopolysaccharide(LPS)-induced inflammatory injury in rat intestinal epithelial(IEC-6)cells,as well as the underlying mechanisms.An in vitro inflam-mation model was first established by treating IEC-6 cells with lipopolysaccharide(LPS).IEC-6 cells were then incubated with three concentrations of TFHL for 24 h prior to a further 24 h LPS treatment.RT-qPCR was used to quantify mRNA levels of the inflammatory genes COX-2 and iN-OS,while Western blotting was used to assess protein levels of the apoptotic markers Bax,cleaved Caspase-3,Bcl-2,and the JNK/p-JNK signaling pathway.Finally,cells were pretreated with TFHL and/or the JNK inhibitor SP600125 for 24 h before LPS exposure for 24 h,in order to evaluate the combined effects of TFHL and SP600125 on LPS-induced inflammatory cytokine expression and apoptotic protein levels in IEC-6 cells.The results showed that,compared with the LPS group,the mRNA level of COX-2 and iNOS in the 2.5,5.0,10.0 mg/L TFHL group and the Bax and Caspase-3 protein levels decreased significantly(P<0.01),and the Bcl-2 protein level was significantly higher(P<0.01),p-JNK protein level and p-JNK/JNK ratio decreased significantly(P<0.01);compared with the LPS group,the COX-2 and iNOS mRNA levels of the TFHL+LPS group de-creased significantly(P<0.01),Bax,and Caspase-3 protein levels decreased significantly(P<0.01),and the level of Bcl-2 protein increased significantly(P<0.05);compared with the LPS group,the COX-2 and iNOS mRNA levels of the TFHL+SP600125 group decreased significantly(P<0.01),Bax and Caspase-3 protein levels decreased significantly(P<0.01),and Bcl-2 protein level increased significantly(P<0.01).These findings indicate that TFHL exerts anti-inflammato-ry and anti-apoptotic effects in LPS-challenged IEC-6 cells by inhibiting the JNK signaling path-way.
7.A review of animal models of non-specific lower back pain
Qiang ZHANG ; Baoqiang DONG ; Xingxing LIN ; Ziwei LIU ; Leichao WANG ; Feng ZHANG ; Danning ZHANG ; Kaixuan ZHANG
Acta Laboratorium Animalis Scientia Sinica 2024;32(12):1594-1605
Non-specific lower back pain is a common clinical disease whose pathogenesis and causes are still unclear,and the advantages and disadvantages of various therapeutic programs are controversial.Current research on this disease is mostly limited to clinical studies,and there is an urgent need to invest in a large number of animal experiments to analyze its underlying mechanisms.The construction of animal models is an important means to study the pathogenesis of non-specific lower back pain and to explore therapeutic method,but there are certain limitations and delays in the establishment of models for this disease.Therefore,this paper reviews the selection of animals,construction method,and evaluation method of relevant indexes of animal models of non-specific lower back pain,and the advantages and disadvantages of various models,to clarify existing problems in current basic research of this disease.We provide new research ideas and aim to lay a theoretical foundation for studies into the mechanisms of non-specific lower back pain and improved therapeutic strategies.
8.Should patients with suspected breast implant-associated anaplastic large cell lymphoma be tested for T-cell receptor gene rearrangement?
Yuxin LIU ; Jiaming SUN ; Jiajia LIU ; Cen QIU ; Junqi CUI ; Danning ZHENG ; Li YU
Chinese Journal of Plastic Surgery 2024;40(5):514-519
Breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) is a rare type of T-cell lymphoma. Despite the scarcity of reported BIA-ALCL cases in Asia, it is imperative to research early diagnosis. The crucial diagnostic criteria for BIA-ALCL include the presence of ALK - and CD30 + T cells exceeding 10% in the delayed seroma fluid. Furthermore, laboratory tests, such as histological examination of capsulectomies and analysis of clonal T-cell receptor (TCR) gene rearrangements, serve as important auxiliary diagnostic indicators. This article reported the case of a 56-year-old female patient who underwent bilateral breast augmentation with implants over 20 years ago. She presented with hardness, enlargement, and mild discomfort in her left breast. She was admitted to Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine in January 2023. MRI suggested implant rupture. Therefore, bilateral implant removal surgery was performed on February 2, 2023. Pathological examination of the fluid within the capsule of the left implant revealed a small number of ALK - and CD30 + T cells, with monoclonality observed in TCRγ gene rearrangement, indicating early changes suggestive of BIA-ALCL. Long-term follow-up is needed. The authors suggest that patients suspected of BIA-ALCL should undergo TCR gene rearrangement testing in addition to cytological and immunological examinations, which can provide guidance for the diagnosis, treatment, and necessary long-term follow-up of these patients.
9.Should patients with suspected breast implant-associated anaplastic large cell lymphoma be tested for T-cell receptor gene rearrangement?
Yuxin LIU ; Jiaming SUN ; Jiajia LIU ; Cen QIU ; Junqi CUI ; Danning ZHENG ; Li YU
Chinese Journal of Plastic Surgery 2024;40(5):514-519
Breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) is a rare type of T-cell lymphoma. Despite the scarcity of reported BIA-ALCL cases in Asia, it is imperative to research early diagnosis. The crucial diagnostic criteria for BIA-ALCL include the presence of ALK - and CD30 + T cells exceeding 10% in the delayed seroma fluid. Furthermore, laboratory tests, such as histological examination of capsulectomies and analysis of clonal T-cell receptor (TCR) gene rearrangements, serve as important auxiliary diagnostic indicators. This article reported the case of a 56-year-old female patient who underwent bilateral breast augmentation with implants over 20 years ago. She presented with hardness, enlargement, and mild discomfort in her left breast. She was admitted to Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine in January 2023. MRI suggested implant rupture. Therefore, bilateral implant removal surgery was performed on February 2, 2023. Pathological examination of the fluid within the capsule of the left implant revealed a small number of ALK - and CD30 + T cells, with monoclonality observed in TCRγ gene rearrangement, indicating early changes suggestive of BIA-ALCL. Long-term follow-up is needed. The authors suggest that patients suspected of BIA-ALCL should undergo TCR gene rearrangement testing in addition to cytological and immunological examinations, which can provide guidance for the diagnosis, treatment, and necessary long-term follow-up of these patients.
10.Clinical features and gene detection analysis in a family with congenital aniridia
Journal of Army Medical University 2024;46(11):1277-1283
Objective To analyze the genetic features of congenital aniridia in a family and to explore the characteristics of PAX6 gene mutations and differences in clinical phenotypes.Methods The medical history and clinical data of this family line were collected.Whole-exon gene sequencing and data analysis were performed,and homology modelling server SWISS-MODEL was applied to construct the corresponding protein model for analysis.Results All 4 individuals with onset in this family line had photophobia and difficulty in opening the eyes due to iris deficiency,with the same clinical phenotypes of aniridia,cataracts,and macular centro-concave dysplasia.But,there were still individual differences in phenotypes.Genetic examination showed that all 4 affected individuals were heterozygous for a heterozygous deletion shifted variant c.442_452del:p.M148Afs*48 on the PAX6 gene,which is highly conserved among species.Homozygous modeling suggested that the mutated PAX6 gene ultimately led to differential changes in protein conformation.Conclusion The newly identified PAX6 gene variant is not phenotypically identical among members of the family with congenital aniridia,and homology modeling analysis has increased the understanding that aberrant expression of PAX6 leads to structural abnormalities in the protein.Genetic examination may provide genetic evidence for this family line.

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