1.Experimental study on effect of circ-DCAF7 on proliferation and migration of prostate cancer cells by regulating miRNA-18a-5p
Jia LIU ; Yunfei ZHAO ; Xiaoying WANG ; Fang XIE ; Geng HUANG ; Hong WANG
Cancer Research and Clinic 2025;37(9):641-647
Objective:To investigate the relationship between circular RNA (circRNA) circ-DCAF7 and the survival of prostate cancer patients, as well as the effects and potential mechanisms of circ-DCAF7 on proliferation and migration of prostate cancer cells in vitro.Methods:The expression and survival data of circ-DCAF7 gene were obtained from 151 prostate cancer patients using the PROGgeneV2 online platform. Patients were divided into circ-DCAF7 high and low expression groups based on the median expression level of circ-DCAF7 gene, and the difference in overall survival between the two groups was compared. Real-time fluorescence quantitative polymerase chain reaction (qPCR) was used to detect the expression of circ-DCAF7 at transcription level in human prostate cancer cell lines 22Rv1, VCaP, PC-3, DU-145, LNCaP, and human normal prostate epithelial cell line RWPE-1. The control plasmid (carrying irrelevant control sequences) and the plasmid carrying circ-DCAF7 sequences were transfected into VCaP cells using liposome reagents, respectively, and referred to as the control group and circ-DCAF7 group. The proliferation and migration abilities of two groups of VCaP cells were detected by plate clone formation assay and cell scratch assay, respectively; using CircInteractome online software, specific binding sites were predicted between circ-DCAF7 and miRNA-18a-5p (miR-18a-5p) sequences, and validated using dual luciferase reporter gene assay; qPCR was used to detect the expression of miR-18a-5p at transcription level in VCaP cells of control group and circ-DCAF7 group; Western blotting was used to detect the expression levels of AKT signaling pathway factors p-AKT, p-mTOR, HIF-1α, and c-myc proteins in two groups of VCaP cells.Results:Analysis using the PROGgeneV2 online tool showed that the overall survival of prostate cancer patients in the circ-DCAF7 high expression group was better than that in the circ-DCAF7 low expression group, and the difference was statistically significant ( P < 0.001). Compared with the normal prostate epithelial cell line RWPE-1, the relative expression of circ-DCAF7 at transcription level in all prostate cancer cell lines was lower, and the differences were statistically significant (all P < 0.05). Among them, the expression level of circ-DCAF7 in VCaP cells was the lowest ( P < 0.001). The relative expression of circ-DCAF7 at transcription level in VCaP cells in the circ-DCAF7 group was higher than that in the control group (9.88±1.58 vs. 1.04±0.39), and the difference was statistically significant ( t = 5.42, P = 0.002). The number of VCaP cell clone formation in the circ-DCAF7 group was less than that in the control group (35±12 vs. 116±11), and the difference was statistically significant ( t = 4.96, P = 0.003). The 25-hour scratch healing rate of VCaP cells in the circ-DCAF7 group was lower than that in the control group [(16±3)% vs. (52±6)%], and the difference was statistically significant ( t = 5.37, P = 0.002). The dual luciferase reporter gene assay showed that compared with VCaP cells co-transfected with miR-18a-5p unrelated control and carrying wild-type circ-DCAF7 sequence plasmid, the relative luciferase activity of VCaP cells co-transfected with miR-18a-5p and carrying wild-type circ-DCAF7 sequence plasmid was lower, and the difference was statistically significant ( t = 7.31, P < 0.001). The relative expression of miR-18a-5p at transcription level in VCaP cells in the circ-DCAF7 group was lower than that in the control group (0.99±0.15 vs. 7.55±1.12), and the difference was statistically significant ( t = 5.83, P = 0.001). Western blotting analysis showed that the expression levels of p-AKT, p-mTOR, HIF-1α, and c-myc proteins of the AKT signaling pathway in VCAP cells of the circ-DCAF7 group were lower than those in the control group. Conclusions:The low expression level of circRNA circ-DCAF7 may be associated with poor prognosis of prostate cancer patients; upregulation of circ-DCAF7 level in prostate cancer cells may inhibit cell proliferation and migration by regulating the miR-18a-5p/AKT signaling pathway.
2.Current status and development suggestions for acupuncture standards.
Ziyi CHONG ; Bo GAO ; Yunfei XIE ; Lirong JIA
Chinese Acupuncture & Moxibustion 2025;45(11):1650-1660
This study organizes and classifies acupuncture-related standards, and analyzes the current status and existing problems. At present, there are 63 international standards, 40 national standards, 6 industry standards, 43 local standards, and 194 group standards related to acupuncture. The current situation is characterized by an irrational standard structure and incomplete coverage of relevant fields; insufficient coordination mechanisms, with overlapping and redundant standards; slow updating of standards, lacking timeliness. It is suggested that the development of acupuncture-related standards should be promoted through the following measures: improving the structure of the standard system and establishing a sound collaborative management mechanism; implementing full-cycle management of standards in line with technological advancements; and building an acupuncture standards information platform to provide one-stop services.
Acupuncture Therapy/standards*
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Humans
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Acupuncture/standards*
3.Experimental study on effect of circ-DCAF7 on proliferation and migration of prostate cancer cells by regulating miRNA-18a-5p
Jia LIU ; Yunfei ZHAO ; Xiaoying WANG ; Fang XIE ; Geng HUANG ; Hong WANG
Cancer Research and Clinic 2025;37(9):641-647
Objective:To investigate the relationship between circular RNA (circRNA) circ-DCAF7 and the survival of prostate cancer patients, as well as the effects and potential mechanisms of circ-DCAF7 on proliferation and migration of prostate cancer cells in vitro.Methods:The expression and survival data of circ-DCAF7 gene were obtained from 151 prostate cancer patients using the PROGgeneV2 online platform. Patients were divided into circ-DCAF7 high and low expression groups based on the median expression level of circ-DCAF7 gene, and the difference in overall survival between the two groups was compared. Real-time fluorescence quantitative polymerase chain reaction (qPCR) was used to detect the expression of circ-DCAF7 at transcription level in human prostate cancer cell lines 22Rv1, VCaP, PC-3, DU-145, LNCaP, and human normal prostate epithelial cell line RWPE-1. The control plasmid (carrying irrelevant control sequences) and the plasmid carrying circ-DCAF7 sequences were transfected into VCaP cells using liposome reagents, respectively, and referred to as the control group and circ-DCAF7 group. The proliferation and migration abilities of two groups of VCaP cells were detected by plate clone formation assay and cell scratch assay, respectively; using CircInteractome online software, specific binding sites were predicted between circ-DCAF7 and miRNA-18a-5p (miR-18a-5p) sequences, and validated using dual luciferase reporter gene assay; qPCR was used to detect the expression of miR-18a-5p at transcription level in VCaP cells of control group and circ-DCAF7 group; Western blotting was used to detect the expression levels of AKT signaling pathway factors p-AKT, p-mTOR, HIF-1α, and c-myc proteins in two groups of VCaP cells.Results:Analysis using the PROGgeneV2 online tool showed that the overall survival of prostate cancer patients in the circ-DCAF7 high expression group was better than that in the circ-DCAF7 low expression group, and the difference was statistically significant ( P < 0.001). Compared with the normal prostate epithelial cell line RWPE-1, the relative expression of circ-DCAF7 at transcription level in all prostate cancer cell lines was lower, and the differences were statistically significant (all P < 0.05). Among them, the expression level of circ-DCAF7 in VCaP cells was the lowest ( P < 0.001). The relative expression of circ-DCAF7 at transcription level in VCaP cells in the circ-DCAF7 group was higher than that in the control group (9.88±1.58 vs. 1.04±0.39), and the difference was statistically significant ( t = 5.42, P = 0.002). The number of VCaP cell clone formation in the circ-DCAF7 group was less than that in the control group (35±12 vs. 116±11), and the difference was statistically significant ( t = 4.96, P = 0.003). The 25-hour scratch healing rate of VCaP cells in the circ-DCAF7 group was lower than that in the control group [(16±3)% vs. (52±6)%], and the difference was statistically significant ( t = 5.37, P = 0.002). The dual luciferase reporter gene assay showed that compared with VCaP cells co-transfected with miR-18a-5p unrelated control and carrying wild-type circ-DCAF7 sequence plasmid, the relative luciferase activity of VCaP cells co-transfected with miR-18a-5p and carrying wild-type circ-DCAF7 sequence plasmid was lower, and the difference was statistically significant ( t = 7.31, P < 0.001). The relative expression of miR-18a-5p at transcription level in VCaP cells in the circ-DCAF7 group was lower than that in the control group (0.99±0.15 vs. 7.55±1.12), and the difference was statistically significant ( t = 5.83, P = 0.001). Western blotting analysis showed that the expression levels of p-AKT, p-mTOR, HIF-1α, and c-myc proteins of the AKT signaling pathway in VCAP cells of the circ-DCAF7 group were lower than those in the control group. Conclusions:The low expression level of circRNA circ-DCAF7 may be associated with poor prognosis of prostate cancer patients; upregulation of circ-DCAF7 level in prostate cancer cells may inhibit cell proliferation and migration by regulating the miR-18a-5p/AKT signaling pathway.
4.Mechanism of total salvianolic acid improving myeloid macrophage infiltration in liver cancer bearing mice by regulating TLR4/MyD88/NF-κB pathway
Yunan DU ; Zhibo DANG ; Xiqian ZHANG ; Gaofeng TAN ; Honglin LI ; Yunfei JIA
Chinese Journal of Immunology 2025;41(11):2624-2630
Objective:To investigate the impacts of total salvianolic acid(TSA)on macrophage polarization and mesenchymal macrophage infiltration in liver cancer bearing mice by regulating the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)pathway.Methods:Mice were randomly separated into liver cancer group,TSA low concentra-tion group,TSA medium concentration group,TSA high concentration group,TSA high concentration+TLR4 inhibitor(TAK242)group and control group,with 12 mice in each group.Except for control group,mice in all other groups were injected subcutaneously into the right axilla to construct liver cancer models by intraperitoneal transmission of third-generation H22 cell suspension.After suc-cessful modeling,administration was carried out once a day for 2 weeks.Changes in tumor weight,tumor volume,spleen index and thymus index were detected.Immunofluorescence was applied to detect proportions of CD86 and CD206 positive cells in tumor tissue.ELISA was applied to detect levels of inducible nitric oxide synthase(iNOS),IL-6 and IL-10 in tumor tissues.Flow cytometry was ap-plied to detect proportion of CD11b+F4/80+in tumor tissue.Western blot was applied to detect TLR4,p-NF-κB P65 and MyD88 pro-teins in tumor tissue.The life quality and survival rate of mice were observed.Results:Compared with control group,spleen index,thymus index,proportion of CD86 positive cells in tumor tissue,levels of iNOS and IL-6,and expressions of TLR4,p-NF-κB P65 and MyD88 proteins were reduced in liver cancer group,while proportion of CD206 positive cells and level of IL-10 in tumor tissue were increased(P<0.05).Compared with liver cancer group,tumor weight and volume of mice in TSA low,medium,high concentration groups were decreased,while survival rate,spleen index,thymus index,proportion of CD86 positive cells in tumor tissue,levels of iNOS and IL-6,and expressions of TLR4,p-NF-κB P65 and MyD88 proteins were increased,proportion of CD206 positive cells,level of IL-10,and proportion of CD11b+F4/80+in tumor tissue decreased(P<0.05).TAK242 reversed the effects of high concentration TSA on macrophage polarization and myeloid macrophage infiltration in liver cancer bearing mice.Conclusion:TSA may promote M1 polari-zation of macrophages in liver cancer bearing mice and inhibit myeloid macrophage infiltration by activating the TLR4/MyD88/NF-κB pathway.
5.Research and Application of Traditional Chinese Medicine Knowledge Organization System
Ziyi CHONG ; Yunfei XIE ; Bo GAO ; Lirong JIA
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(9):1258-1266
The knowledge organization work in the field of Traditional Chinese Medicine(TCM)aims to establish a systematic TCM knowledge system,contributing to the discovery,sharing,and application of knowledge resources.From the perspective of knowledge organization,this paper classifies the TCM knowledge system based on technological development generations and degrees of structurali-zation.Seven representative and distinctive knowledge organization systems in the TCM field were selected,including encyclopedias,dictionaries,classification schemes,subject headings,ontologies,semantic networks,and knowledge graphs.The characteristics of each knowledge system are identified and future research directions are analyzed.The progress of knowledge organization application re-search relies on the support of knowledge organization systems.This paper elaborates on the application scope of TCM knowledge organ-ization systems from three aspects:literature indexing and retrieval,clinical decision support,and intelligent knowledge service con-struction.Finally,it emphasizes that knowledge organization models will develop toward achieving multimodal,fine-grained knowledge organization and advanced reasoning applications,continuously improving the TCM knowledge organization system and fully exploiting its application potential.
6.Mechanism of total salvianolic acid improving myeloid macrophage infiltration in liver cancer bearing mice by regulating TLR4/MyD88/NF-κB pathway
Yunan DU ; Zhibo DANG ; Xiqian ZHANG ; Gaofeng TAN ; Honglin LI ; Yunfei JIA
Chinese Journal of Immunology 2025;41(11):2624-2630
Objective:To investigate the impacts of total salvianolic acid(TSA)on macrophage polarization and mesenchymal macrophage infiltration in liver cancer bearing mice by regulating the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)pathway.Methods:Mice were randomly separated into liver cancer group,TSA low concentra-tion group,TSA medium concentration group,TSA high concentration group,TSA high concentration+TLR4 inhibitor(TAK242)group and control group,with 12 mice in each group.Except for control group,mice in all other groups were injected subcutaneously into the right axilla to construct liver cancer models by intraperitoneal transmission of third-generation H22 cell suspension.After suc-cessful modeling,administration was carried out once a day for 2 weeks.Changes in tumor weight,tumor volume,spleen index and thymus index were detected.Immunofluorescence was applied to detect proportions of CD86 and CD206 positive cells in tumor tissue.ELISA was applied to detect levels of inducible nitric oxide synthase(iNOS),IL-6 and IL-10 in tumor tissues.Flow cytometry was ap-plied to detect proportion of CD11b+F4/80+in tumor tissue.Western blot was applied to detect TLR4,p-NF-κB P65 and MyD88 pro-teins in tumor tissue.The life quality and survival rate of mice were observed.Results:Compared with control group,spleen index,thymus index,proportion of CD86 positive cells in tumor tissue,levels of iNOS and IL-6,and expressions of TLR4,p-NF-κB P65 and MyD88 proteins were reduced in liver cancer group,while proportion of CD206 positive cells and level of IL-10 in tumor tissue were increased(P<0.05).Compared with liver cancer group,tumor weight and volume of mice in TSA low,medium,high concentration groups were decreased,while survival rate,spleen index,thymus index,proportion of CD86 positive cells in tumor tissue,levels of iNOS and IL-6,and expressions of TLR4,p-NF-κB P65 and MyD88 proteins were increased,proportion of CD206 positive cells,level of IL-10,and proportion of CD11b+F4/80+in tumor tissue decreased(P<0.05).TAK242 reversed the effects of high concentration TSA on macrophage polarization and myeloid macrophage infiltration in liver cancer bearing mice.Conclusion:TSA may promote M1 polari-zation of macrophages in liver cancer bearing mice and inhibit myeloid macrophage infiltration by activating the TLR4/MyD88/NF-κB pathway.
7.Research and Application of Traditional Chinese Medicine Knowledge Organization System
Ziyi CHONG ; Yunfei XIE ; Bo GAO ; Lirong JIA
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(9):1258-1266
The knowledge organization work in the field of Traditional Chinese Medicine(TCM)aims to establish a systematic TCM knowledge system,contributing to the discovery,sharing,and application of knowledge resources.From the perspective of knowledge organization,this paper classifies the TCM knowledge system based on technological development generations and degrees of structurali-zation.Seven representative and distinctive knowledge organization systems in the TCM field were selected,including encyclopedias,dictionaries,classification schemes,subject headings,ontologies,semantic networks,and knowledge graphs.The characteristics of each knowledge system are identified and future research directions are analyzed.The progress of knowledge organization application re-search relies on the support of knowledge organization systems.This paper elaborates on the application scope of TCM knowledge organ-ization systems from three aspects:literature indexing and retrieval,clinical decision support,and intelligent knowledge service con-struction.Finally,it emphasizes that knowledge organization models will develop toward achieving multimodal,fine-grained knowledge organization and advanced reasoning applications,continuously improving the TCM knowledge organization system and fully exploiting its application potential.
8.Environmental hygiene of metro line X in Nanjing in 2023
Qiuyuan ZHU ; Zhaojuan XIAN ; Yanzhao TANG ; Yunfei JIA ; Ming GE ; Lilin XIONG
Journal of Environmental and Occupational Medicine 2024;41(9):1054-1060
Background The subway environment, characterized by dense crowds, relatively enclosed architectural structures, and insufficient ventilation, poses various health hazards. Population health risk attributable to subway hygiene have become an important public health issue. Objective To evaluate the current environmental hygiene of metro line X in Nanjing, and to provide a scientific basis for the prevention and control of harmful factors and the establishment of relevant standards. Methods Station halls, platforms, and carriages of Nanjing metro line X were monitored in March 2023. Six stations were selected by stratified sampling, and monitored once during the morning peak hours (9:00—11:00). Carriage monitoring was conducted after a full day of operation (21:00—22:00). The monitoring indicators included physical factors such as temperature, relative humidity, wind speed, illuminance, and noise (A-weighted); air quality factors such as carbon dioxide (CO2), carbon monoxide (CO), respirable particulate matter (PM10), fine particulate matter (PM2.5), ammonia (NH3), formaldehyde, benzene, toluene, xylene, total volatile organic compounds (TVOC), ozone (O3), total bacterial count, and total fungi count; hygiene status of the inner surface of the central air conditioning ventilation system, such as dust accumulation, total bacterial count, and total fungi count; supply air, such as PM10, PM2.5, total bacterial count, and total fungi count; and surface microorganisms on objects, such as total bacterial count, total fungi count, coliform bacteria, and Staphylococcus aureus. Results were evaluated and discussed according to Hygienic indicators and limits for public places (GB 37488-2019) and Hygienic specification of central air conditioning ventilation systems in public buildings (WS 394-2012). Results The environmental temperature medians (P25, P25) in the platforms and carriages of Nanjing metro line X were 20.7 (19.95, 21.65) ℃ and 21.8 (19.80, 35.80) °C, respectively, which exceeded the standard limits. The relative humidity and noise level in the carriages were 38.80% (24.6%, 46.3%) and 79.50 (76.25, 82.00) dB, respectively, failing to meet the national health standards. The total fungi count in supply air in 22.2% of the monitoring points exceeded the standard value. Coliform bacteria and Staphylococcus aureus were detected on the surfaces of some high-touch objects, with the bathroom faucet being the most severely contaminated. The medians (P25, P25) of temperature [21.8 (19.8, 35.8) ℃], wind speed [0.48 (0.39, 1.02) m·s−1], noise [79.5 (76.25, 82.00) dB, and CO2 content [0.079% (0.070%, 0.091%)] in the carriages were higher than those in the station halls and platforms (P<0.05). There were significant differences in microclimate, air particulate matter, and air microbial levels between urban and suburban stations (P<0.05); the concentrations of PM10 [0.076 (0.046, 0.079) mg·m−3)] and PM2.5 [0.063 (0.044, 0.068) mg·m−3)] in suburban stations were 2 times higher than those of urban stations. The temperature [21.60 (20.45, 21.80) ℃], humidity [45.20% (40.95%, 50.10%)], CO2 [0.100% (0.100%, 0.825%)], and PM10 concentration [0.070 (0.041, 0.080) mg·m−3] in transfer stations were all significantly higher than those in non-transfer stations (P<0.05). The temperature at stations was significantly correlated with the CO2 content (rs=
9.Challenges and suggestions for human genetic resource management in international cooperation of medical institutions caused by the Implementation Rules of the Regulations on the Management of Human Genetic Resources: A case study of Peking University Cancer Hospital
Shuanglei KONG ; Yunfei GENG ; Luopei WEI ; Lingling BAI ; Shuqin JIA
Chinese Journal of Medical Science Research Management 2024;37(1):18-21
Objective:According to the international cooperation project of Peking University Cancer Hospital on human genetic resource management practices, combined with the development direction of human genetic resource management laws and regulations, and propose reference suggestions for medical institutions to strengthen human genetic resource management.Methods:Sort out the projects that Peking University Cancer Hospital obtained international cooperation approval on the government platform of the Ministry of Science and Technology from July 2019 to June 2023, analyze the current situation of human genetic resource management in the hospital, summarize the challenges brought by the implementation of new regulations on human genetic resource management in medical institutions, and propose corresponding suggestions.Results:A total of 1276 international cooperation projects on human genetic resources have been approved, including 345 initial declarations and 931 change declarations. Involving 453 studies, including 286 clinical trials of drugs or devices on the market, accounting for 63.13%, and 100 clinical trials of Phase I drugs, accounting for 34.97% of the market studies. On average, there are 3.14 changes per project for listed research, and 1.56 changes per project for non listed research.Conclusions:Regulations on the Management of Human Genetic Resources ( short for Rules) limit the management scope of international cooperation projects involving human genetic resources and delegate management authority to medical institutions. Adjusting the scope of application for international cooperative clinical trial filing may result in some administrative approval projects being transferred to filing. The approval process for international cooperative scientific research projects on human genetic resources has been adjusted. Suggest medical institutions to strengthen the management of samples and intellectual property outside the scope of application of Rules.Strengthen the entire process management of international cooperation in scientific research. Pay attention to and timely communicate the dynamics of human genetic resource management.
10.Screening for Characteristic Genes of Different Traditional Chinese Medicine Syndromes of Psoriasis Vulgaris:A Study Based on Bioinformatics and Machine Learning
Xuewei LIU ; Huangchao JIA ; Liyun WANG ; Ziwen WANG ; Mengyue XU ; Yunfei LI ; Ronghui WANG
Journal of Sichuan University (Medical Sciences) 2024;55(2):337-345
Objective To screen for the key characteristic genes of the psoriasis vulgaris(PV)patients with different Traditional Chinese Medicine(TCM)syndromes,including blood-heat syndrome(BHS),blood stasis syndrome(BSS),and blood-dryness syndrome(BDS),through bioinformatics and machine learning and to provide a scientific basis for the clinical diagnosis and treatment of PV of different TCM syndrome types.Methods The GSE192867 dataset was downloaded from Gene Expression Omnibus(GEO).The limma package was used to screen for the differentially expressed genes(DEGs)of PV,BHS,BSS,and BDS in PV patients and healthy populations.In addition,KEGG(Kyoto Encyclopedia of Genes and Genes)pathway enrichment analysis was performed.The DEGs associated with PV,BHS,BSS,and BDS were identified in the screening and were intersected separately to obtain differentially characterized genes.Out of two algorithms,the support vector machine(SVM)and random forest(RF),the one that produced the optimal performance was used to analyze the characteristic genes and the top 5 genes were identified as the key characteristic genes.The receiver operating characteristic(ROC)curves of the key characteristic genes were plotted by using the pROC package,the area under curve(AUC)was calculated,and the diagnostic performance was evaluated,accordingly.Results The numbers of DEGs associated with PV,BHS,BSS,and BDS were 7 699,7 291,7 654,and 6 578,respectively.KEGG enrichment analysis was focused on Janus kinase(JAK)/signal transducer and activator of transcription(STAT),cyclic adenosine monophosphate(cAMP),mitogen-activated protein kinase(MAPK),apoptosis,and other pathways.A total of 13 key characteristic genes were identified in the screening by machine learning.Among the 13 key characteristic genes,malectin(MLEC),TUB like protein 3(TULP3),SET domain containing 9(SETD9),nuclear envelope integral membrane protein 2(NEMP2),and BTG anti-proliferation factor 3(BTG3)were the key characteristic genes of BHS;phosphatase 15(DUSP15),C1q and tumor necrosis factor related protein 7(C1QTNF7),solute carrier family 12 member 5(SLC12A5),tripartite motif containing 63(TRIM63),and ubiquitin associated protein 1 like(UBAP1L)were the key characteristic genes of BSS;recombinant mouse protein(RRNAD1),GTPase-activating protein ASAP3 Protein(ASAP3),and human myomesin 2(MYOM2)were the key characteristic genes of BDS.Moreover,all of them showed high diagnostic efficacy.Conclusion There are significant differences in the characteristic genes of different PV syndromes and they may be potential biomarkers for diagnosing TCM syndromes of PV.

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