1.Genetic diversity analysis and DNA fingerprinting of Artemisia argyi germplasm resources based on EST-SSR molecular markers.
Yu-Yang MA ; Chang-Jie CHEN ; Ming-Xing WANG ; Yan FANG ; Yu-Huan MIAO ; Da-Hui LIU
China Journal of Chinese Materia Medica 2025;50(9):2356-2364
This study investigates the genetic diversity and evolutionary relationships of different Artemisia argyi germplasm resources to provide a basis for germplasm identification, variety selection, and resource protection. A total of 192 germplasm resources of A. argyi were studied, and EST-based simple sequence repeat(EST-SSR) primers were designed based on transcriptomic data of A. argyi. Polymerase chain reaction(PCR) amplification was performed on these resources, followed by fluorescence capillary electrophoresis to detect genetic diversity and construct DNA fingerprints. From 197 pairs of primers designed, 28 pairs with polymorphic and clear bands were selected. A total of 278 alleles were detected, with an average of 9.900 0 alleles per primer pair and an average effective number of alleles of 1.407 2. The Shannon's diversity index(I) for the A. argyi germplasm resources ranged from 0.148 1 to 0.418 0, with an average of 0.255 7. The polymorphism information content(PIC) ranged from 0.454 5 to 0.878 0, with an average of 0.766 9, showing high polymorphism. Cluster analysis divided the A. argyi germplasm resources into three major groups: Group Ⅰ contained 136 germplasm samples, Group Ⅱ contained 45, and Group Ⅲ contained 11. Principal component analysis also divided the resources into three groups, which was generally consistent with the clustering results. Mantel test results showed that the genetic variation in A. argyi populations was to some extent influenced by geographic distance, but the effect was minimal. Structure analysis showed that 190 germplasm materials had Q≥ 0.6, indicating that these germplasm materials had a relatively homogeneous genetic origin. Furthermore, 8 core primer pairs were selected from the 28 designed primers, which could distinguish various germplasm types. Using these 8 core primers, DNA fingerprints for the 192 A. argyi germplasm resources were successfully constructed. EST-SSR molecular markers can be used to study the genetic diversity and phylogenetic relationships of A. argyi, providing theoretical support for the identification and molecular-assisted breeding of A. argyi germplasm resources.
Artemisia/classification*
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Microsatellite Repeats
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Genetic Variation
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Expressed Sequence Tags
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DNA Fingerprinting
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Phylogeny
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Polymorphism, Genetic
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DNA, Plant/genetics*
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Genetic Markers
2.Swyer syndrome with gonadal non-dysgerminoma malignant germ cell tumors: a report of 15 cases in a national medical center.
Huan LIANG ; Si Jie LI ; Jia Xin YANG ; Ming WU ; Dong Yan CAO ; Jin Hui WANG ; Tao WANG ; Xin Yue ZHANG
Chinese Journal of Obstetrics and Gynecology 2024;59(1):64-69
Objective: To evaluate the incidence, treatment, and survival outcomes of Swyer syndrome with gonadal non-dysgerminoma malignant germ cell tumor (MGCT-NDG). Methods: A retrospective study was performed on Swyer syndrome patients with MGCT-NDG between January 2011 and December 2022 in Peking Union Medical College Hospital to investigate their characteristics and outcomes. Results: A total of 15 patients (4.9%, 15/307) with Swyer syndrome were identified in 307 MGCT-NDG patients. The average age at diagnosis of MGCT-NDG and Swyer syndrome were (16.8±6.7) and (16.7±6.6) years, respectively. Six cases were preoperatively diagnosed as Swyer syndrome, of which 4 cases received bilateral gonadectomy with or without hysterectomy, while the other 2 cases underwent removal of gonadal tumor and unilateral gonadectomy with hysterectomy, respectively. Of the 9 patients postoperatively diagnosed as Swyer syndrome, unilateral gonadectomy, removal of gonadal tumor, and unilateral gonadectomy with hysterectomy were performed in 6 patients, 2 patients, and 1 patient, respectively. Mixed malignant germ cell tumor (MGCT;10 cases), yolk sac tumor (4 cases), and immature teratoma (1 case) were the pathological subtypes, in the descending order. There were International Federation of Gynecology and Obstetrics (FIGO) stage Ⅰ in 6 cases, stage Ⅱ in 3 cases, stage Ⅲ in 5 cases, and stage Ⅳ in 1 case, respectively. Eleven patients received reoperation for residual gonadectomy after a average delay of (7.9±6.2) months, including 8 MGCT-NDG patients and 1 gonadoblastoma patient, no tumor involved was seen in the remaining gonads in the other 2 cases. Ten patients experienced at least one recurrence, with a median event free survival of 9 months (5, 30 months), of which 2 patients received surgery only at the time of initial treatment. All patients with recurrence received surgery and combined with postoperative chemotherapy. After a median follow-up of 25 months (15, 42 months), 10 patients were disease-free, 3 patients died of the tumor, 1 died of side effects of leukemia chemotherapy, and 1 survived with disease. Conclusion: The incidence rate of Swyer syndrome in patients with MGCT-NDG is about 4.9%; timely diagnosis and bilateral gonadectomy should be emphasized to reduce the risk of reoperation and second carcinogenesis in this population.
Female
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Humans
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Retrospective Studies
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Gonadal Dysgenesis, 46,XY/surgery*
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Gonadoblastoma/surgery*
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Neoplasms, Germ Cell and Embryonal/surgery*
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Ovarian Neoplasms/pathology*
3.Repeatability of wavefront aberration measured by adaptive optics visual simulator and agreement of OPD-Scan Ⅲ
Yan WANG ; Xuan LIAO ; Changjun LAN ; Biao LI ; Zhuang MIAO ; Qingqing TAN ; Suyun QIN ; Huan HUANG
International Eye Science 2024;24(5):810-815
AIM: To assess the repeatability and agreement of higher-order aberration obtained by adaptive optics visual simulator(VAO)compared with OPD-Scan Ⅲ.METHODS: A cross-sectional study was conducted from August to September 2023, including a total of 204 patients(204 eyes)with myopia whose right eyes were measured. The examinations were performed by the same skilled examiner using both devices separately. The VAO device was used to measure higher order aberrations of orders 3 to 6 at a pupil diameter of 4.5 mm, while both the VAO and OPD-Scan Ⅲ devices were utilized to measure total higher-order aberration(tHOA), spherical aberration(SA), coma aberration(Coma), and trefoil aberration(Trefoil)of the entire eye at pupil diameters ranging from 3 to 6 mm. Furthermore, the repeatability of whole eye aberration measurements obtained with the VAO device was evaluated and the agreement of the two devices was assessed.RESULTS: The whole-eye higher-order aberrations measured by VAO demonstrated excellent repeatability(0.767≤ICC≤0.941, Sw<0.01 μm, TRT<0.1 μm). There was no statistically significant difference in Coma measured by VAO or OPD-Scan Ⅲ for pupil diameters ranging from 4 to 6 mm(P>0.05), while a statistically significant difference was observed in whole-eye tHOA of other pupil diameters(all P<0.05). The agreement of aberration measurements for each order between VAO and OPD-Scan Ⅲ for 3 mm pupil diameters, SA at 4 and 5 mm pupil diameter and Coma at 4 mm pupil diameter showed a 95% limit of agreement(LoA)<0.1, indicating good agreement; however, poor agreement was found for the remaining aberration measurements at different pupil diameters, with a 95%LoA>0.1, and there were significant differences in higher-order aberrations measured by two devices under a pupil diameter of 3 mm(r=0.218-0.317, P<0.01), 4 mm(r=0.406-0.672, P<0.01), 5 mm(r=0.538-0.839, P<0.01 and r=0.030-0.109, P>0.01)and 6 mm(r=0.369-0.766, P<0.01).CONCLUSION: The VAO demonstrates favorable repeatability when assessing whole-eye higher order aberration under pupil diameters of 3-6 mm. However, there is inadequate agreement and interchangeability in whole-eye higher order aberration at 3-6 mm pupil diameter between VAO and OPD-Scan Ⅲ for clinical purposes.
4.Prediction analysis of quality markers and resource evaluation of Artemisiae Argyi Folium based on chemical composition and network pharmacology.
Chang-Jie CHEN ; Hong-Zhi DU ; Yu-Huan MIAO ; Yan FANG ; Ting-Ting ZHAO ; Da-Hui LIU
China Journal of Chinese Materia Medica 2023;48(20):5474-5486
This study is based on ultra-high-performance liquid chromatography(UPLC), gas chromatography-mass spectrometry(GC-MS), and network pharmacology methods to analyze and predict potential quality markers(Q-markers) of Artemisiae Argyi Folium. First, UPLC and GC-MS techniques were used to analyze the content of 12 non-volatile components and 8 volatile components in the leaves of 33 Artemisia argyi germplasm resources as candidate Q-markers. Subsequently, network pharmacology was employed to construct a "component-target-pathway-efficacy" network to screen out core Q-markers, and the biological activity of the markers was validated using molecular docking. Finally, cluster analysis and principal component analysis were performed on the content of Q-markers in the 33 A. argyi germplasm resources. The results showed that 18 candidate components, 60 targets, and 185 relationships were identified, which were associated with 72 pathways related to the treatment of 11 diseases and exhibited 5 other effects. Based on the combination of freedom and component specificity, six components, including eupatilin, cineole, β-caryophyllene, dinatin, jaceosidin, and caryophyllene oxide were selected as potential Q-markers for Artemisiae Argyi Folium. According to the content of these six markers, cluster analysis divided the 33 A. argyi germplasm resources into three groups, and principal component analysis identified S14 as having the highest overall quality. This study provides a reference for exploring Q-markers of Artemisiae Argyi Folium, establishing a quality evaluation system, further studying its pharmacological mechanisms, and breeding new varieties.
Molecular Docking Simulation
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Network Pharmacology
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Plant Breeding
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Chromatography, High Pressure Liquid/methods*
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Gas Chromatography-Mass Spectrometry
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Artemisia/chemistry*
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Drugs, Chinese Herbal/chemistry*
5.Signal transducer and activator of transcription 3 and cancer associated fibroblasts jointly generate chemo-resistance and affect prognosis in epithelial ovarian cancer.
Ya Nan ZHANG ; Bin LI ; Yu Qing LI ; Shuang Huan LIU ; Hong Yi HOU ; Kun Yu WANG ; Miao AO ; Yan SONG
Chinese Journal of Obstetrics and Gynecology 2023;58(5):368-377
Objective: To investigate the mechanism of signal transducer and activator of transcription 3 (STAT3) and cancer associated fibroblasts (CAF) jointly generate chemo-resistance in epithelial-ovarian cancer and their effect on prognosis. Methods: A total of 119 patients with high-grade ovarian serous cancer who received surgery in Cancer Hospital of Chinese Academy of Medical Sciences from September 2009 to October 2017 were collected. The clinico-pathological data and follow-up data were complete. Multivariate Cox regression model was used to analyze the prognostic factors. Ovarian cancer tissue chips of patients in our hospital were prepared. EnVision two-step method immunohistochemistry was used to detect the protein expression levels of STAT3, the specific markers of CAF activation, fibroblast activating protein (FAP), and type Ⅰ collagen (COL1A1) secreted by CAF. The relationship between the expression of STAT3, FAP, COL1A1 protein and drug resistance and prognosis of ovarian cancer patients was analyzed, and the correlation between the expression of three proteins was analyzed. These results were verified through the gene expression and prognostic information of human ovarian cancer tissues collected in the GSE26712 dataset of gene expression omnibus (GEO) database. Results: (1) Multivariate Cox regression model analysis showed that chemotherapy resistance was an independent risk factor for overall survival (OS) of ovarian cancer (P<0.001). (2) The expression levels of STAT3, FAP, and COL1A1 proteins in chemotherapy resistant patients were significantly higher than those in chemotherapy sensitive patients (all P<0.05). Patients with high expression of STAT3, FAP, and COL1A1 had significantly shorter OS than those with low expression (all P<0.05). According to the human ovarian cancer GSE26712 dataset of GEO database, patients with high expression of STAT3, FAP, and COL1A1 also showed shorter OS than patients with low expression (all P<0.05), the verification results were consistent with the detection results of ovarian cancer patients in our hospital. (3) Correlation analysis showed that the protein level of STAT3 was positively correlated with FAP and COL1A1 in our hospital's ovarian cancer tissue chips (r=0.47, P<0.001; r=0.30, P=0.006), the analysis of GEO database GSE26712 dataset showed that the expression of STAT3 gene and FAP, COL1A1 gene were also significantly positively correlated (r=0.31, P<0.001; r=0.52, P<0.001). Conclusion: STAT3 and CAF could promote chemotherapy resistance of ovarian cancer and lead to poor prognosis.
Female
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Humans
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Cancer-Associated Fibroblasts/pathology*
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Carcinoma, Ovarian Epithelial
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Ovarian Neoplasms/pathology*
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Prognosis
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STAT3 Transcription Factor/metabolism*
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Drug Resistance, Neoplasm
6.Content of mineral elements in different Artemisia argyi germplasms and their relationship with quality properties.
Chang-Jie CHEN ; Yu-Huan MIAO ; Yan FANG ; Lan-Ping GUO ; Yuan ZENG ; Da-Hui LIU
China Journal of Chinese Materia Medica 2022;47(4):880-888
To clarify the content characteristics of mineral elements in different Artemisia argyi germplasm resources and their relationship with the quality properties of Artemisiae Argyi Folium, this study measured the content of 10 mineral elements including nitrogen(N), phosphorus(P), potassium(K), calcium(Ca), magnesium(Mg), aluminum(Al), manganese(Mn), iron(Fe), copper(Cu), and zinc(Zn) in 100 Artemisia argyi germplasm samples. Besides, their relationship with the quality properties of Artemisiae Argyi Folium was explored by correlation analysis, path analysis, and cluster analysis. The results demonstrated that the variation coefficient of the 10 mineral elements in Artemisiae Argyi Folium ranged from 12.23% to 64.38%, and the genetic diversity index from 0.97 to 3.09. The genetic diversities of N, P, and Zn were obvious. As revealed by the correlation analysis, N, P, and K showed strong positive correlations with each other. Except that Mg and Al were negatively correlated, Ca, Mg, Al, Mn, Fe, Cu, and Zn were positively correlated. The correlation analysis of mineral elements with the quality properties of Artemisiae Argyi Folium proved the significant correlations of 17 pairs of characters. According to the path analysis, P, K, Ca, and Mn greatly affected the yield of Artemisiae Argyi Folium, P, K, and Mg the output rate of moxa, N, P, and K the content of total volatile oil, P and K the content of eucalyptol, and P, K, and Ca the content of eupatilin. The 100 germplasm samples were clustered into three groups. Specifically, in cluster Ⅰ, the enrichment capacity of P, K, and Mg elements was strong, and the comprehensive properties of mineral elements were better, implying good development potential. Ca, Mn, Fe, and Zn elements in cluster Ⅱ and N and Al in cluster Ⅲ displayed strong enrichment capacities. This study has provided new ideas for resource evaluation and variety breeding of A. argyi and also reference for fertilizer application.
Artemisia/genetics*
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Iron
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Minerals/analysis*
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Plant Breeding
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Plant Leaves/chemistry*
8.Lrrc34 Is Highly Expressed in SSCs and Is Necessary for SSC Expansion In Vitro.
Jin Huan OU ; Yi Ran LI ; Zhi Peng WANG ; Cheng JIN ; Kai LI ; Yan LU ; Ding Feng ZOU ; Peng Yu LI ; Meng Zhen LI ; Shi Ying MIAO ; Lin Fang WANG ; Wei SONG
Chinese Medical Sciences Journal 2020;35(1):20-30
Objective To discover critical genes contributing to the stemness and maintenance of spermatogonial stem cells (SSCs) and provide new insights into the function of the leucine-rich repeat (LRR) family member Lrrc34 (leucine-rich repeat-containing 34) in SSCs from mice. Methods Bioinformatic methods, including differentially expressed gene (DEG), gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, were used to uncover latent pluripotency-related genes. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence analyses were utilized to verify the mRNA and protein expression levels, respectively. RNA interference of Lrrc34 using siRNA was performed to detect its transient impact on SSCs. Results Eight DEGs between ID4-EGFP+ (G) and ID4-EGFP+/TSPAN8High (TH), eight DEGs between G and ID4-EGFP+/TSPAN8Low (TL) and eleven DEGs between TH and TL were discovered, and eleven protein-protein interaction (PPI) modules were found to be significant in the PPI network of DEGs. One of the DEGs, Lrrc34, was selected as a potential pluripotency-related gene due to its differential expression among ID4-EGFP+ spermatogonia subsets and its interaction with fibroblast growth factor 2 in the fifth module. Immunofluorescence experiments exhibited specific expression of Lrrc34 in a subpopulation of undifferentiated spermatogonia marked by LIN28A, and RT-PCR experiments confirmed the high expression of Lrrc34 in SSCs from P7 and adult mice. The transient knockdown of Lrrc34 in SSCs resulted in reduced colony sizes and significant changes in the transcriptome and apoptotic pathways. Conclusion Lrrc34 is highly expressed in mouse SSCs and is required for SSC proliferation in vitro through effects on transcriptome and signaling transduction pathways.
Animals
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Apoptosis/genetics*
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Cell Proliferation/genetics*
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Cells, Cultured
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Gene Expression Profiling/methods*
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Gene Ontology
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Gene Regulatory Networks
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Humans
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Male
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Mice, Inbred C57BL
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Mice, Transgenic
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RNA Interference
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Repressor Proteins/metabolism*
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Signal Transduction/genetics*
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Stem Cells/metabolism*
9.Prediction of the relationship between the mandibular and craniofacial bone and cervical vertebrae based on a genetic algorithm in patients with skeletal class Ⅱ malocclusion
MU Qili ; YANG Luyi ; ZHAO Xuejiao ; YAN Jing ; YU Miao ; WANG Liyao ; WANG Huan
Journal of Prevention and Treatment for Stomatological Diseases 2019;27(11):711-717
Objective:
To explore the correlation between the parameters of the mandible and parameters of cervical vertebrae and craniofacial bone in class Ⅱ skeletal patients in Northeast China and to establish correlation equations expressing the relationship between the mandible and cervical vertebrae and craniofacial bone directly and quantitatively for the clinical diagnosis and treatment of orthodontics and orthognathics and for prediction.
Methods :
The mandible, cranial facial bone and cervical vertebrae of 201 children and adolescents aged 8 to 20 years were measured using digital cranial lateral tablets. All of the cases were divided into male (n=75) and female (n=126) groups using a sensitivity analysis method based on genetic algorithms to select the craniofacial bone and cervical bone with strong sensitivity to mandible parameters and to establish relevant equations.
Results :
Through sensitivity analysis, the parameters with the strongest correlation between the measured values of the mandible were H4 and SN, those with a strong correlation were SN-Ar, the anterior and posterior high ratio SGo/NGn, the Y axis angle and mandibular angle Ar-Go-Gn. The established equation was as follows: males: Ar-Pg=28.415+1.818×H4+0.746×SN(r2=0.056 8, P < 0.001); females: Ar-Pg=15.168+1.706×H4+0.675×SN+0.31×SN-Ar-0.29×Y axis angle (r2=0.611, P < 0.001). No significant difference was found between the predicted values obtained by the established equations and measured values (P > 0.05).
Conclusion
The mandibular length equation established by sensitivity analysis and genetic algorithms is statistically significant and can predict a certain degree of growth and development.
10.Predictive values of the platelet to lymphocyte ratio and neutrophil to lymphocyte ratio for drug eluting stent restenosis
Yang ZHENG ; Xiao-Huan LIU ; Wei-Dong MA ; Shan JIA ; Ya-Jie FAN ; Zhi-Hui YAO ; Chun-Yan ZHANG ; Yan ZHANG ; Yan-Chao HU ; Miao GE ; Cong-Xia WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2018;39(4):466-470
Objective To investigate the predictive values of platelet to lymphocyte ratio (PLR) and neutrophil to lymphocyte ratio (NLR)before percutaneous coronary intervention (PCI)and a reexamination of coronary angiography (CAG)on the development of in-stent restenosis (ISR)in patients implanted with coronary drug eluting stent (DES).Methods For this study we enrolled 123 patients who had undergone successful drug eluting stent implantation (SI)and a further CAG reexamination.Another 45 patients with non-coronary heart disease (NC)served as controls.PLR and NLR were measured before DES implantation or CAG and compared between the groups.Patients in SI group were further divided into two subgroups based on the results of CAG reexamination:ISR group and no-ISR group.Hematologic data were reexamined before further CAG and compared between the subgroups.Receiver operator characteristic curve (ROC)was drawn to evaluate the predictive values of PLR and NLR for ISR.Results PLR and NLR before PCI or a further CAG were significantly higher in ISR group (34 patients)than in non-ISR group (89 patients,P<0.05).Before PCI,the best cutoff value of PLR in screening restenosis was 107.20;the sensitivity and the specificity were 64.7% and 65.2%.The best cutoff value of NLR in screening restenosis was 2.72; the sensitivity and the specificity were 61.8% and 70.8%. Before CAG reexamination,the best cutoff value of PLR in screening restenosis was 160.08;the sensitivity and the specificity were 26.5% and 97.8%.The best cutoff value of NLR in screening restenosis was 2.08;the sensitivity and the specificity were 73.5% and 56.2%.Conclusion Both PLR and NLR before PCI or CAG reexamination can be predictors of ISR in patients implanted with DES.


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