1.Mechanism of Yantiao Prescription in Treating Lipopolysaccharide-induced Acute Lung Injury Based on Arachidonic Acid Metabolic Pathways
Pengcheng LI ; Tianyang CHEN ; Rong FANG ; Anna ZHANG ; Sijia WU ; Wei LIU ; Qian WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):101-110
ObjectiveTo clarify the anti-inflammatory and lung-protective effects of Yantiao prescription on lipopolysaccharide (LPS)-induced acute lung injury (ALI), and to explore the impact of Yantiao prescription on the metabolic pathways of arachidonic acid (AA) in vivo. MethodsThirty male C57BL/6J mice were randomly divided into the following groups based on body weight: normal group, model group, dexamethasone group (2 mg·kg-1), low-dose Yantiao prescription group (18 g·kg-1), and high-dose Yantiao prescription group (36 g·kg-1), with 6 mice in each group. The ALI mouse model was established by intraperitoneal injection of LPS. The treatment groups received oral gavage once a day for 7 consecutive days, and serum and lung tissue were collected at the end of the experiment. The content of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in serum was detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining was used to assess lung tissue pathology. The wet/dry weight ratio (W/D) and myeloperoxidase (MPO) activity in lung tissue were measured. The content of AA metabolites in serum and lung tissue was measured by liquid chromatography triple quadrupole-mass spectrometry (LC-MS/MS). ResultsCompared with the conditions in the normal group, the content of serum pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in the model group was significantly increased (P<0.01). The alveolar structure in mice was severely damaged, with markedly thickened alveolar walls and extensive inflammatory cell infiltration. The W/D ratio and MPO activity in lung tissue were significantly increased (P<0.01). The content of AA metabolites, including prostaglandin D2 (PGD2), prostaglandin E2 (PGE2), 11(S)-hydroxy-eicosatetraenoic acid [11(S)-HETE], and 5-hydroxy-eicosatetraenoic acid (5-HETE) in serum and lung tissue was significantly increased (P<0.05), while the content of 11,12-epoxyeicosatrienoic acid (11,12-EET) and 14,15-epoxyeicosatrienoic acid (14,15-EET) in serum was significantly decreased (P<0.01). Compared with the results in the model group, the content of serum pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in the dexamethasone group, low-dose Yantiao prescription group, and high-dose Yantiao prescription group was significantly reduced (P<0.05). Mild thickening of alveolar walls, scattered inflammatory cell infiltration, and relatively intact tissue structure with improved alveolar architecture were observed. The W/D ratio and MPO activity in lung tissue were significantly reduced (P<0.01). The content of AA metabolites PGD2, PGE2, 11(S)-HETE, and 5-HETE in serum from the dexamethasone group was significantly decreased (P<0.05), while the content of 14,15-EET in serum significantly increased (P<0.01), and the content of 5-HETE in lung tissue significantly decreased (P<0.01). In the low-dose and high-dose Yantiao prescription groups, the content of AA metabolites PGD2, PGE2, 11(S)-HETE, and 5-HETE in serum and lung tissue was significantly decreased (P<0.05), while the content of 11,12-EET in both serum and lung tissue was significantly increased (P<0.05). ConclusionYantiao prescription has significant protective effects against LPS-induced ALI, which are related to its regulation of AA metabolic pathways in vivo.
2.Herbal Textual Research on Picrorhizae Rhizoma in Famous Classical Formulas
Feng ZHOU ; Yihan WANG ; Yanmeng LIU ; Xiaoqin ZHAO ; Kaizhi WU ; Cheng FENG ; Wenyue LI ; Wei ZHANG ; Wentao FANG ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):228-239
This article systematically analyzes the historical evolution of the name, origin, quality evaluation, harvesting, processing and other aspects of Picrorhizae Rhizoma by referring to the medical books, prescription books, and other documents of the past dynasties, combined with relevant modern research materials, in order to provide a basis for the development and utilization of famous classical formulas containing this medicinal herb. The research results indicate that Picrorhizae Rhizoma was first recorded in New Revised Materia Medica from the Tang dynasty. Throughout history, Huhuanglian has been used as its official name, and there are also aliases such as Gehu Luze, Jiahuanglian and Hulian. The main source of past dynasties is the the rhizomes of Picrorhiza kurrooa and P. scrophulariiflora. In ancient times, Picrorhizae Rhizoma was mainly imported by foreign traders via Guangzhou and other regions, and also produced in China, mainly in Xizang. In ancient times, it was harvested and dried in early August of the lunar calendar, while in modern times, it is mostly harvested from July to September, with the best quality being those with thick and crispy rhizomes without impurities, and bitter taste. Throughout history, Picrorhizae Rhizoma was collected, washed, sliced, and dried before being used as a raw material for medicine, it has a bitter and cold taste, mainly used to treat bone steaming, hot flashes, infantile chancre fever, and dysentery. There is no significant difference in taste and efficacy between ancient and modern times. Based on the research results, it is recommended that the rhizomes of P. scrophulariiflora in the 2020 edition of Chinese Pharmacopoeia, or the rhizomes of P. kurrooa, can be used in famous classical formulas containing this medicinal herb, which can be processed according to the processing requirements marked by the original formula. For those without clear processing requirements, the dried raw products are used as medicine.
3.Mechanism of Astragaloside Ⅳ in Regulating PI3K/Akt Molecular Pathway in Prevention and Treatment of Diabetes Complications: A Review
Kexin HU ; Jinru ZHU ; Qing WU ; Shengmao WANG ; Mengfan WANG ; Ai QIAN ; Zhaohui FANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):307-314
Diabetes mellitus (DM) is a metabolic disease caused by absolute or relative insulin deficiency and reduced insulin sensitivity in peripheral cells, posing a serious threat to global health. Chronic complications arising in the later stages of DM can lead to the decline or even loss of function in multiple organs, including the eyes, heart, liver, kidneys, nerves, and feet, making them the primary cause of mortality in DM patients. Although modern medicine has made some progress in the treatment of these complications, challenges such as high costs and adverse drug reactions remain. Thus, identifying highly effective drugs with minimal adverse effects has become a top priority. Astragalus membranaceus is a shining gem in the treasure trove of Chinese medicine. Numerous studies have shown that its primary active component, astragaloside Ⅳ, possesses various biological activities, including anti-inflammatory, antioxidant, and antiviral effects, as well as benefits for cardiac and cerebral function, nerve conduction, and myocardial protection. Meanwhile, the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway plays a crucial role in regulating oxidative stress, inflammatory responses, apoptosis, and autophagy. Extensive research has highlighted the significant role of this pathway in various DM complications, leading to widespread studies on its interaction with astragaloside Ⅳ. This review summarizes research findings on how astragaloside Ⅳ alleviates pancreatic cytotoxicity in DM patients by modulating the PI3K/Akt pathway. Additionally, it highlights its protective effects on basic cardiac function, inhibition of retinal cell damage, improvement of cerebral nerve dysfunction, reduction of chronic kidney and liver damage, and mitigation of neurovascular toxicity in the lower limbs. These insights provide a valuable reference for the clinical application of A. membranaceus and its active monomer, astragaloside Ⅳ, in the treatment of DM and its complications.
4.Surveillance results of respiratory syncytial virus outbreaks in kindergarten and school in Shenzhen, 2017-2023
WANG Xin, FANG Shisong, WU Weihua, LIU Hui, SUN Ying, ZOU Xuan, TANG Xiujuan
Chinese Journal of School Health 2025;46(3):435-437
Objective:
To analyze respiratory syncytial virus(RSV) outbreaks surveillance results and the epidemiological characteristics in kindergarten and school in Shenzhen during 2017-2023 , so as to provide a scientific reference for control and prevention of RSV.
Methods:
Epidemiological data and surveillance results of RSV outbreaks in kindergarten and school from 2017 to 2023 were collected for descriptive analyses.
Results:
A total of 31 RSV outbreaks were identified in kindergarten and school in 2017-2023 in Shenzhen, 346 cases were reported, the average incidence rate was 22.02%. The most annual RSV outbreaks were reported in 2020 with 14 outbreaks, followed by 8 outbreaks in 2023. A total of 64.52% of RSV outbreaks were identified in kindergarten with rest occurring in primary school or middle school. The greatest monthly count of outbreak was 18 (58.06%) in September, followed by 3 outbreaks (9.68%) in March and October. A total of 244 swab samples were collected, 169 samples were positive for respiratory viruses, the positive rate was 69.26%, 121 samples were positive for RSV,from 31 respiratory syncytical virus outbreaks 57 and samples were positive for other respiratory viruses(9 samples were positive for two respiratory viruses). A toral of 14(45.16%) outbreaks are caused by RSV alone, 17 outbreaks (54.84%) were caused by RSV and other respiratory viruses.
Conclusions
Most RSV outbreaks in kindergarten and school are reported after 2020 in Shenzhen, most RSV outbreaks occur in kindergarten, peak seasons of RSV outbreaks are autumn and spring.
5.Analysis of Potential Active Components and Molecular Mechanism of Baoxin Granules Regulating Ferroptosis in Treatment of Heart Failure
Yu CHEN ; Maolin WANG ; Yun WANG ; Yifan ZHAO ; Jing XU ; Hongwei WU ; Fang WANG ; Xiaoang ZHAO ; Youming LI ; Jixiang TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):202-209
ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), network pharmacology, molecular docking and cell experiments, the active ingredients, possible targets and molecular mechanisms of Baoxin granules(BXG) regulating ferroptosis in the treatment of heart failure(HF) were explored. MethodsBXG intestinal absorption fluid was prepared by everted gut sac and the chemical composition contained therein were identified by UPLC-Q-TOF-MS. According to the obtained components, the potential targets of BXG were predicted, and the HF-related targets and related genes of ferroptosis were retrieved at the same time, and the intersecting targets were obtained by Venn diagram. In addition, the protein-protein interaction(PPI) network and the component-target network were constructed, and the core components and core targets were obtained by topological analysis. Then Gene Ontology(GO) function and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis were performed on the core targets, and molecular docking validation of the key targets and main components was carried out by AutoDockTools 1.5.7. H9c2 cells were used to establish a oxygen-glucose deprivation model, and the protective effect of BXG on cells was investigated by detecting cell viability, cell survival rate and reactive oxygen species(ROS) level. The protein expression levels of signal transducer and activator of transcription 3(STAT3), phosphorylation(p)-STAT3 and glutathione peroxidase 4(GPX4) were detected by Western blot to clarify the regulatory effect of BXG on ferroptosis. ResultsA total of 61 chemical components in BXG intestinal absorption fluid were identified, and network pharmacology obtained 27 potential targets of BXG for the treatment of HF, as well as 139 signaling pathways. BXG may act on core targets such as STAT3, tumor protein p53(TP53), epidermal growth factor receptor(EGFR), JUN and prostaglandin-endoperoxide synthase 2(PTGS2) through core components such as glabrolide and limonin, which in turn intervene in lipid and atherosclerosis, phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt), endocrine resistance and other signaling pathways to exert therapeutic effects on HF. Molecular docking showed that the docking results of multiple groups of targets and compounds were good. In vitro cell experiments showed that compared with the blank group, the cell viability and survival rate of the model group were significantly decreased, the level of ROS was significantly increased(P<0.01), the expression levels of STAT3, p-STAT3, p-STAT3/STAT3 and GPX4 proteins were significantly decreased(P<0.05, P<0.01). Compared with the model group, the cell viability and survival rate of the BXG group were significantly increased, the ROS level was significantly decreased(P<0.01), the STAT3, p-STAT3, p-STAT3/STAT3 and GPX4 protein levels were significantly increased(P<0.05, P<0.01). ConclusionBXG may inhibit the occurrence of ferroptosis by up-regulating the expression of STAT3 and GPX4, thus exerting a therapeutic effect on HF, and flavonoids may be the key components of this role.
6.Analysis of Potential Active Components and Molecular Mechanism of Baoxin Granules Regulating Ferroptosis in Treatment of Heart Failure
Yu CHEN ; Maolin WANG ; Yun WANG ; Yifan ZHAO ; Jing XU ; Hongwei WU ; Fang WANG ; Xiaoang ZHAO ; Youming LI ; Jixiang TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):202-209
ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), network pharmacology, molecular docking and cell experiments, the active ingredients, possible targets and molecular mechanisms of Baoxin granules(BXG) regulating ferroptosis in the treatment of heart failure(HF) were explored. MethodsBXG intestinal absorption fluid was prepared by everted gut sac and the chemical composition contained therein were identified by UPLC-Q-TOF-MS. According to the obtained components, the potential targets of BXG were predicted, and the HF-related targets and related genes of ferroptosis were retrieved at the same time, and the intersecting targets were obtained by Venn diagram. In addition, the protein-protein interaction(PPI) network and the component-target network were constructed, and the core components and core targets were obtained by topological analysis. Then Gene Ontology(GO) function and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis were performed on the core targets, and molecular docking validation of the key targets and main components was carried out by AutoDockTools 1.5.7. H9c2 cells were used to establish a oxygen-glucose deprivation model, and the protective effect of BXG on cells was investigated by detecting cell viability, cell survival rate and reactive oxygen species(ROS) level. The protein expression levels of signal transducer and activator of transcription 3(STAT3), phosphorylation(p)-STAT3 and glutathione peroxidase 4(GPX4) were detected by Western blot to clarify the regulatory effect of BXG on ferroptosis. ResultsA total of 61 chemical components in BXG intestinal absorption fluid were identified, and network pharmacology obtained 27 potential targets of BXG for the treatment of HF, as well as 139 signaling pathways. BXG may act on core targets such as STAT3, tumor protein p53(TP53), epidermal growth factor receptor(EGFR), JUN and prostaglandin-endoperoxide synthase 2(PTGS2) through core components such as glabrolide and limonin, which in turn intervene in lipid and atherosclerosis, phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt), endocrine resistance and other signaling pathways to exert therapeutic effects on HF. Molecular docking showed that the docking results of multiple groups of targets and compounds were good. In vitro cell experiments showed that compared with the blank group, the cell viability and survival rate of the model group were significantly decreased, the level of ROS was significantly increased(P<0.01), the expression levels of STAT3, p-STAT3, p-STAT3/STAT3 and GPX4 proteins were significantly decreased(P<0.05, P<0.01). Compared with the model group, the cell viability and survival rate of the BXG group were significantly increased, the ROS level was significantly decreased(P<0.01), the STAT3, p-STAT3, p-STAT3/STAT3 and GPX4 protein levels were significantly increased(P<0.05, P<0.01). ConclusionBXG may inhibit the occurrence of ferroptosis by up-regulating the expression of STAT3 and GPX4, thus exerting a therapeutic effect on HF, and flavonoids may be the key components of this role.
7.Study on the stability of PBMCs recovered from leukocyte-depleted filter residues
Ju LIN ; Zhiqiang XIANG ; Dongfen DU ; Fang YUAN ; Miaoyu WANG ; Yue WU ; Kaiyu HUANG ; Lieyong SANG
Chinese Journal of Blood Transfusion 2025;38(5):723-733
Objective: To identify an optimal back-flush solution for leukocyte-depleted filters that maximizes peripheral blood mononuclear cell (PBMC) recovery with high viability, long-term storage stability, and sterility of the harvested residues, thereby providing a clinically translatable strategy. Methods: Three sterile bag-packaged solutions—Saline, Solvent, and Hanks' balanced salt solution (HBSS)—were used to back-flush randomly assigned leukocyte-depleted filters. Nucleated cell recovery rate and viability of the harvested residues were compared. The optimal solution identified was applied to an expanded sample set. PBMC viability and yield were evaluated after 1h vs 48h storage of the residues. PBMCs isolated from the residues were cryopreserved in liquid nitrogen for 1 month, followed by post-thaw comparisons of viability and T-cell expansion capacity. Results: The Solvent group achieved the highest and most consistent nucleated cell recovery rate. Post-flush recovery rate from filters after 400 mL whole blood processing was (21.3±1.6)% for the Solvent group, significantly higher than Saline group (19.2±6.3)% and HBSS group (11.2±5.0)%, with residues from all groups maintaining viability >90%. No biologically significant difference in residue viability was observed between 48h vs 1h storage groups (93.3±2.3)% vs (95.7±1.8)%). PBMC recovery rates from residues showed no statistical difference between 48h vs 1h storage groups [(48.2%±9.5%)vs (40.41%±8.35%), P>0.05], with (17.7±2.6)×10
cells. After 1-month cryopreservation and 10-day expansion, PBMCs isolated from 48-hour-stored residues retained (91.2±3.2)% viability and achieved a (61.9±15.9)-fold expansion. Conclusion: The bag-packaged Solvent, as a back-flush solution, enables sterile acquisition of leukocyte-depleted filter residues through closed-system tubing connections. These residues maintained PBMC viability and recovery rates after 48h storage at 2℃-8℃, with post-cryopreservation (1-month liquid nitrogen) viability and expansion capacity remaining stable. This protocol complies with blood bank regulatory criteria, addresses the concerns about the infectious window period in cell therapy raw materials, and provides a clinically translatable strategy for PBMC-based applications.
8.Results and analysis of inter-comparison of gross α and β measurement capabilities in Gansu Province, China, 2023
Jialong WU ; Fang WANG ; Yanjun WANG ; Qin CHEN
Chinese Journal of Radiological Health 2025;34(2):249-253
Objective To assess the quality of gross α and β radioactivity measurements conducted by radiological health service institutions and disease prevention and control centers in Gansu Province, China, and regulate their measurement methods. Methods The samples were distributed by Gansu Provincial Center for Disease Control and Prevention as the organizer of the inter-comparison through mail. The institutions participated in the inter-comparison carried out the measurements in accordance with national standards, and submitted the inter-comparison reports in the form required by the inter-comparison scheme. Results A total of 13 institutions participated in the 2023 inter-comparison of gross α and β radioactivity measurement capabilities, and all measurement results met the required standards. The absolute Z-scores for gross α inter-comparison ranged from 0 to 1.21, and the absolute Z-scores for gross β inter-comparison ranged from 0.08 to 1.85. The comprehensive scores ranged from 74.5 to 93.0. Conclusion The measurement capacities of the institutions participated in the 2023 inter-comparison showed improvement compared with the previous year. However, 12 institutions participated in the inter-comparison showed issues in data processing, report formatting, and laboratory quality control. It is necessary to strengthen technical training, standardize the measurement procedures, and improve the measurement capabilities and skills to ensure the quality of services.
9.Degradation dynamics and residue determination of pyriclobenzuron in rice and paddy environments
Huanqi WU ; Junmin WANG ; Kongtan YANG ; Xumi WANG ; Nan FANG ; Liping DUAN ; Changpeng ZHANG ; Xiangyun WANG
Chinese Journal of Schistosomiasis Control 2025;37(2):201-208
Objective To establish a method for determination of pyriclobenzuron (PBU) residues in rice and paddy environments, and to determine the residual amounts and observe the degradation dynamics of PBU. Methods In July 2022, the paddies of Zhejiang Academy of Agricultural Sciences were selected as experimental fields, and were divided into the blank control group (no pesticide application), the 1-fold-concentration pesticide group (1 kg/667 m2), and the 5-fold-concentration pesticide group (5 kg/667 m2), with a 100 m2 area in each group. At the early tillering stage of rice, 20% suspension of PBU sulfate was sprayed once in the 1-fold-concentration and 5-fold-concentration pesticide groups, and rice plants, paddy water and soil samples were collected 2 h, and 1, 2, 3, 5, 7, 11, 14, 21, 28, 35, 49 d and 63 d following spraying PBU, while rice straw, field soil, brown rice and rice husk samples were collected 98 d following spraying. PBU was extracted and purified in samples using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) pretreatment technique, and the PBU contents were determined in samples using ultrahigh performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The solvent standard working solution and matrix standard working solution were prepared. A linear regression equation was fitted between PBU concentration (x-axis) and peak area (y-axis), and the ratio of the slope (k) of the matrix standard curve to the slope (K) of the solvent standard curve was calculated to evaluate the matrix effect of PBU in samples. According to the Guidelines for Pesticide Residue Testing in Crops (NY/T 788—2018), the addition levels of PBU were set at 0.005, 0.050, 5.000, 1 000.000 mg/kg in rice plants, 0.005, 0.050, 2.000, 10.000 mg/kg in paddy water, 0.005, 0.050, 2.000 mg/kg in soil, and 0.005, 0.050, 5.000 mg/kg in brown rice and rice husks. The recovery and relative standard deviation (RSD) of PBU addition were calculated to evaluate the effectiveness of UPLC-MS/MS for determination of PBU contents. The first-order kinetic equation of PBU concentration was fitted in samples at different sampling time points to analyze the trends in PBU degradation in rice plants, paddy water, and soil, and the half-life of PBU was calculated in different samples. Results There was a good linear relationship between the mass concentration and peak area of PBU at concentrations of 0.000 1 to 0.020 0 mg/kg under solvent and matrix conditions (R2 = 0.985 8 to 0.999 7, t = -0.47 to 1.62, all P values < 0.01). The matrix effects of PBU were 70.26%, 65.42% and 65.12% in rice plants, brown rice and rice husks, indicating a matrix-inhibitory effect, and the matrix effect was 87.06% in soils, indicating a weak matrix effect. The recovery of PBU addition was 77.61% to 100.12% in different samples, with RSD of 1.43% to 6.74%, and a limit of quantification (LOQ) of 0.005 mg/kg, and the addition recovery and RSD met the requirements of the Guidelines for Pesticide Residue Testing in Crops (NY/T 788—2018), validating the effectiveness of UPLC-MS/MS assay. Following spraying PBU at a dose of 1 kg/667 m2, the half-life of PBU was 6.24 d in rice plants and 3.43 d in paddy water samples, respectively. The final residues of PBU were lower than the LOQ of 0.005 mg/kg in brown rice and rice husk samples 98 d following spraying PBU. Following spraying PBU at a dose of 5 kg/667 m2, the half-life of PBU was 15.75 d in rice plants and 7.62 d in paddy water samples, respectively. The final residue of PBU was lower than the LOQ of 0.005 mg/kg in brown rice 98 d following spraying PBU, and the final residue of PBU was 0.049 mg/kg in rice husks. Conclusions A simple, and highly accurate and precise UPLC-MS/MS assay has been developed for determination of PBU residues in rice plants and paddy environments through extraction and purification of PBU from matrix samples using QuEChERS pretreatment. After spraying PBU in paddies, the concentration of PBU gradually decreases in rice plants and paddy water over time, and the final residual concentration is low.
10.A whole genome analysis of two coxsackievirus A2 strains isolated from patients with herpetic angina in Shanghai
Jingyi ZHANG ; Jiayu WANG ; Run LI ; Fanghao FANG ; Wencheng WU ; Wanju ZHANG ; Min CHEN ; Xin CHEN ; Zheng TENG
Shanghai Journal of Preventive Medicine 2025;37(3):215-221
ObjectiveTo understand the whole genome characteristics and the information for genetic evolution in the two coxsackievirus A2 (CVA2) strains isolated from patients with herpangina in Shanghai, and to provide a scientific basis for the prevention and treatment of herpetic angina. MethodsTwo CAV2 strains isolated from patients with herpetic angina in Shanghai were performed whole genome sequencing and analysis for phylogenetics, nucleotide homology, and evolution. ResultsA phylogenetic analysis of the VP1 region revealed that the two Shanghai strains both belonged to CVA2 genotype D, with the highest homology to OL357660, a strain from Yunnan. The average nucleotide identity (ANI) of the whole genome between the two Shanghai strains was 98.88%, and the ANI of the whole genome comparisons to other CVA2 genotype D strains and CVA2 genotypes A-C strains ranged from 84.64% to 97.42% and from 79.21% to 84.20%, respectively. The two Shanghai strains had low homology in the 3D region compared to the existing CVA2 strains. The phylogenetic analysis and sliding window nucleotide similarity analysis indicated that the two Shanghai strains and the Yunnan OL357660 strain might constitute a new genetic lineage. ConclusionThe two CVA2 strains isolated for the first time in Shanghai are assigned to genotype D (GenBank: PQ130039 and PQ130040), which is identical to the existing subtype prevalent in China. As represented by the Shanghai strains, a new CVA2 genetic lineage is been identified. This study has enriched the data on genetic evolution and genetic variation of CVA2 in Shanghai, indicating the requirement to strengthen surveillance for the epidemiological pattern of CVA2.


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