1.Tiaowei Jiannao acupuncture for post-ischemic stroke insomnia: a randomized controlled trial.
Run ZHANG ; Xinwang CHEN ; Mengyu WANG ; Wenming CHU ; Lihua WU ; Jing GAO ; Peidong LIU ; Ce SHI ; Liyuan LIU ; Bingzhen LI ; Miaomiao JI ; Yayong HE
Chinese Acupuncture & Moxibustion 2025;45(10):1405-1413
OBJECTIVE:
To observe the efficacy and safety of Tiaowei Jiannao acupuncture (acupuncture for regulating defensive qi and nourishing brain) for post-ischemic stroke insomnia (PISI).
METHODS:
A total of 96 patients with PISI were randomized into an acupuncture group (32 cases, 1 case was excluded), a medication group (32 cases, 1 case dropped out, 1 case was excluded) and a sham-acupuncture group (32 cases, 1 case dropped out, 1 case was excluded). In the acupuncture group, Tiaowei Jiannao acupuncture was applied at bilateral Shenmai (BL62), Zhaohai (KI6), Hegu (LI4), Taichong (LR3), and Baihui (GV20), Sishencong (EX-HN1), Yintang (GV24+), Shenting (GV24), once a day, 1-day interval was taken after 6-day treatment, for 3 weeks totally. In the medication group, eszopiclone tablet was given orally, 1-3 mg a time, once a day for 3 weeks. In the sham-acupuncture group, non-invasive sham acupuncture was applied, the acupoint selection, frequency and course of treatment were the same as the acupuncture group. Before treatment, after 2,3 weeks of treatment, the scores of Pittsburgh sleep quality index (PSQI), self-rating sleep scale (SRSS), National Institutes of Health Stroke scale (NIHSS), Hamilton depression scale-17 (HAMD-17) were observed; before and after treatment, the sleep parameters were recorded using polysomnography (PSG); and the efficacy and safety were evaluated after treatment in the 3 groups.
RESULTS:
After 2,3 weeks of treatment, the scores of PSQI, HAMD-17 and SRSS in the acupuncture group and the medication group, as well as the SRSS scores in the sham-acupuncture group were decreased compared with those before treatment (P<0.05); after 2 weeks of treatment, the NIHSS score in the acupuncture group was decreased compared with that before treatment (P<0.05); after 3 weeks of treatment, the NIHSS scores in the acupuncture group, the medication group and the sham-acupuncture group were decreased compared with those before treatment (P<0.05). After 3 weeks of treatment, the scores of PSQI, SRSS, HAMD-17 and NIHSS in the acupuncture group and the medication group, as well as the NIHSS score in the sham-acupuncture group were decreased compared with those after 2 weeks of treatment (P<0.05). After 2,3 weeks of treatment, the scores of PSQI, SRSS and HAMD-17 in the acupuncture group and the medication group were lower than those in the sham-acupuncture group (P<0.05), the NIHSS scores in the acupuncture group were lower than those in the medication group and the sham-acupuncture group (P<0.05); after 3 weeks of treatment, HAMD-17 score in the acupuncture group was lower than that in the medication group (P<0.05), the NIHSS score in the medication group was lower than that in the sham-acupuncture group (P<0.05). Compared before treatment, after treatment, the total sleep time was prolonged (P<0.05), the wake after sleep onset, sleep latency, and non-rapid eye movement (NREM) sleep latency were shortened (P<0.05), the sleep efficiency was improved (P<0.05), the number of awakenings was reduced (P<0.05), the percentage of rapid eye movement (REM%) and the percentage of NREM stage 1 (N1%) were decreased (P<0.05), the percentage of NREM stage 2 (N2%) and the percentage of NREM stage 3 (N3%) were increased (P<0.05) in the acupuncture group and the medication group; the sleep latency was shortened in the sham-acupuncture group (P<0.05). After treatment, the PSG indexes in the acupuncture group and the medication group were superior to those in the sham-acupuncture group (P<0.05); in the acupuncture group, the number of awakenings was less than that in the medication group (P<0.05), the REM% and N1% were lower than those in the medication group (P<0.05), the N2% and N3% were higher than those in the medication group (P<0.05). The total effective rate were 93.5% (29/31) and 90.0% (27/30) in the acupuncture group and the medication group respectively, which were higher than 10.0% (3/30) in the sham-acupuncture group (P<0.05). There was no serious adverse events in any of the 3 groups.
CONCLUSION
Tiaowei Jiannao acupuncture improves the insomnia symptoms in patients with ischemic stroke, improves the quality of sleep, increases the deep sleep, promotes the recovery of neurological function, and relieves the depression. It is effective and safe for the treatment of PISI.
Humans
;
Acupuncture Therapy
;
Male
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Sleep Initiation and Maintenance Disorders/physiopathology*
;
Female
;
Middle Aged
;
Aged
;
Acupuncture Points
;
Treatment Outcome
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Adult
;
Ischemic Stroke/complications*
;
Stroke/complications*
;
Sleep
2.Graph Neural Networks and Multimodal DTI Features for Schizophrenia Classification: Insights from Brain Network Analysis and Gene Expression.
Jingjing GAO ; Heping TANG ; Zhengning WANG ; Yanling LI ; Na LUO ; Ming SONG ; Sangma XIE ; Weiyang SHI ; Hao YAN ; Lin LU ; Jun YAN ; Peng LI ; Yuqing SONG ; Jun CHEN ; Yunchun CHEN ; Huaning WANG ; Wenming LIU ; Zhigang LI ; Hua GUO ; Ping WAN ; Luxian LV ; Yongfeng YANG ; Huiling WANG ; Hongxing ZHANG ; Huawang WU ; Yuping NING ; Dai ZHANG ; Tianzi JIANG
Neuroscience Bulletin 2025;41(6):933-950
Schizophrenia (SZ) stands as a severe psychiatric disorder. This study applied diffusion tensor imaging (DTI) data in conjunction with graph neural networks to distinguish SZ patients from normal controls (NCs) and showcases the superior performance of a graph neural network integrating combined fractional anisotropy and fiber number brain network features, achieving an accuracy of 73.79% in distinguishing SZ patients from NCs. Beyond mere discrimination, our study delved deeper into the advantages of utilizing white matter brain network features for identifying SZ patients through interpretable model analysis and gene expression analysis. These analyses uncovered intricate interrelationships between brain imaging markers and genetic biomarkers, providing novel insights into the neuropathological basis of SZ. In summary, our findings underscore the potential of graph neural networks applied to multimodal DTI data for enhancing SZ detection through an integrated analysis of neuroimaging and genetic features.
Humans
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Schizophrenia/pathology*
;
Diffusion Tensor Imaging/methods*
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Male
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Female
;
Adult
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Brain/metabolism*
;
Young Adult
;
Middle Aged
;
White Matter/pathology*
;
Gene Expression
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Nerve Net/diagnostic imaging*
;
Graph Neural Networks
3.HPLC Fingerprint and Content Determination of Five Components in Ranunculus Sceleratus L.
Shanshan CAO ; Zhen SHI ; Xiaowen ZHENG ; Fei CHEN ; Sijia ZHOU ; Wenming CHENG ; Yazhong ZHANG ; Jin XIE ; Qunlin ZHANG
Chinese Journal of Modern Applied Pharmacy 2024;41(1):106-111
OBJECTIVE
To establish the HPLC fingerprint and content determination of five components in Ranunculus sceleratus L..
METHODS
The separation was developed on an Agilent ZORBAX SB C18 chromatographic (4.6 mm×250 mm, 5 μm)column by gradient elution with methanol(A)-0.1 % phosphoric acid aqueous solution(B) as mobile phase to establish HPLC fingerprint of Ranunculus sceleratus L.. Combined with similarity evaluation, cluster analysis, principal component analysis, and orthogonal partial least squares-discriminant analysis, the quality of 13 batches of Ranunculus sceleratus L. was evaluated.
RESULTS
Thirteen batches of Ranunculus sceleratus L. samples were calibrated with 20 common peaks, of which 5 common peaks were identified, and the similarity ranged from 0.874 to 0.984. The results of cluster analysis and principal component analysis were basically the same, indicating that there might be differences in the content of chemical components of Ranunculus sceleratus L. in different regions. Protocatechuic aldehyde, caffeic acid, ferulic acid, hyperoside and isoquercitrin were determined in thirteen batches of Ranunculus sceleratus L., and their contents were 0.016−0.035, 0.010−0.070, 0.010−0.029, 0.016−0.051, 0.028−0.086 mg·g–1, respectively.
CONCLUSION
The established HPLC fingerprint and content determination method is simple, stable, and reliable, which can be used for qualitative analysis and provide reference to quality evaluation and resource utilization of Ranunculus sceleratus L..
4.An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique (version 2023)
Jie SHEN ; Lin CHEN ; Shiwu DONG ; Jingshu FU ; Jianzhong GUAN ; Hongbo HE ; Chunli HOU ; Zhiyong HOU ; Gang LI ; Hang LI ; Fengxiang LIU ; Lei LIU ; Feng MA ; Tao NIE ; Chenghe QIN ; Jian SHI ; Hengsheng SHU ; Dong SUN ; Li SUN ; Guanglin WANG ; Xiaohua WANG ; Zhiqiang WANG ; Hongri WU ; Junchao XING ; Jianzhong XU ; Yongqing XU ; Dawei YANG ; Tengbo YU ; Zhi YUAN ; Wenming ZHANG ; Feng ZHAO ; Jiazhuang ZHENG ; Dapeng ZHOU ; Chen ZHU ; Yueliang ZHU ; Zhao XIE ; Xinbao WU ; Changqing ZHANG ; Peifu TANG ; Yingze ZHANG ; Fei LUO
Chinese Journal of Trauma 2023;39(2):107-120
Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.
5.Analysis of risk factors for multiple myeloma combined with radiation-based sarcopenia
Jie REN ; Lei QI ; Xingchen YAO ; Jinzhou WANG ; Xiangjun SHI ; Wenming CHEN ; Xinru DU
Chinese Journal of Orthopaedics 2023;43(9):567-573
Objective:To analyze the risk factors of radiation-based sarcopenia in patients with multiple myeloma (MM).Methods:A total of 185 clinical and imaging data of patients with MM admitted to Beijing Chaoyang Hospital from September 2009 to October 2019 were retrospectively analyzed. The area of the erector spinae muscle and the area of fatty infiltration (FI) in the fascial compartment were measured by Image-pro Ρlus software, and the area of the fat-free erector spinae muscle and the fat infiltration rate (FI%) were calculated. Sarcopenia was defined as an erector spinae area of less than 3 197 mm 2 in males and 2 895 mm 2 in females. The differences in gender, age, body mass index, disease duration, hemoglobin, leukocytes, platelets, albumin, serum calcium, lactate dehydrogenase, serum creatinine, alkaline phosphatase, M-protein, serum β 2-microglobulin, bortezomib chemotherapy, receipt of stem cell transplantation, osteopathy, stage, recurrence and progression of MM between the sarcopenia group and the normal muscle group were compared. Binary logistic regression was used to analyze the independent risk factors of sarcopenia in MM patients. Kaplan-Meier curves were drawn to compare the survival rates between the two groups. Results:53.0% (98/185) of MM patients were complicated with sarcopenia: there were 30 males, whose fat-free erector spinae area was 25.0±6.0 cm 2, the FI of erector spinae was 12.0±4.8 cm 2, and the FI% was 31.5%±12.0%, while there were 68 females, whose fat-free erector spinae area was 22.7±4.2 cm 2, the FI of erector spinae was 10.7±4.1 cm 2, and the FI% was 30.2%±9.8%. 47.0% (87/185) of MM patients had normal muscle mass: there were 62 males, whose fat-free erector spinae area was 40.6±6.5 cm 2, the FI of erector spinae was 9.3±4.8 cm 2, and the FI% was 17.9%±7.4%, while there were 25 females, whose fat-free erector spinae area was 33.6±5.1 cm 2, the FI of erector spinae was 9.9±3.0 cm 2, and the FI% was 21.9%±5.7%. There were statistically significant differences in the gender composition ratio (χ 2=30.47, P<0.001), hemoglobin ( t=-2.73, P=0.007), serum creatinine ( Z=-2.26, P=0.024), receipt of stem cell transplantation (χ 2=4.32, P=0.038), and MM recurrence and progression (χ 2=3.85, P=0.050) between the two groups. However, there were no significant differences in age, body mass index, course of disease, leukocytes, platelets, albumin, alkaline phosphatase, lactate dehydrogenase, serum calcium, M-protein, serum β 2-microglobulin, bortezomib chemotherapy, osteopathy or MM stage ( P>0.05). Binary logistic regression analysis showed that female was an independent risk factor for sarcopenia in MM patients. The survival rates at 2, 3, 4, and 5 years were 87.9%, 71.8%, 64.4%, and 53.7% in the sarcopenia group, and 92.1%, 75.8%, 66.8%, and 66.8% in the normal muscle group, respectively, with no statistically significant differences ( HR=0.71, P=0.364). Conclusion:The incidence of radiation-based sarcopenia in MM patients is 53.0%. Low hemoglobin and blood creatinine levels, not receiving stem cell transplantation, and recurrence or progression of MM are associated with sarcopenia in MM patients, and female is an independent risk factor for sarcopenia in MM patients.
6.Effects of electroacupuncture on Golgi apparatus stress in rats with endotoxin-induced acute lung injury
Wenming LIU ; Jia SHI ; Xiangyun LI ; Yuan ZHANG ; Shu′an DONG ; Yan GUO ; Gerile WUYUN ; Jianbo YU
Chinese Journal of Anesthesiology 2022;42(10):1238-1242
Objective:To evaluate the effect of electroacupuncture (EA) on Golgi apparatus stress in the rats with endotoxin-induced acute lung injury (ALI).Methods:Twenty clean-grade male Sprague-Dawley rats, aged 2 months, weighing 160-185 g, were divided into 4 groups ( n=5 each) according to a random number table method: control group (C group), endotoxin group (LPS group), EA plus endotoxin group (EA+ LPS group), and sham EA plus endotoxin group (SEA+ LPS group).The model of endotoxin-induced ALI was developed by intravenous injection of lipopolysaccharide (LPS) 5 mg/kg in anesthetized animals.Bilateral Zusanli (ST36) and Neiguan (PC6) acupoints were stimulated with an electric stimulator for 30 min once a day at 1-4 days before and during model preparation in group EA+ LPS.In group SEA+ LPS, acupuncture needles were inserted to the surface of ST36 and PC6 acupoints with no current stimulation, and the other parameters were the same as those previously described in group EA+ LPS.Blood samples were collected from the abdominal aorta at 6 h after development of the model for measurement of concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum by enzyme-linked immunosorbent assay.The animals were sacrificed and lungs were removed for microscopic examination of the pathological changes of lung tissues (with a light microscope) and morphological changes of Golgi apparatus (with a transmission electron microscope) and for determination of wet to dry lung weight (W/D) ratio, cell apoptosis index (by TUNEL), activity of superoxide dismutase (SOD) (by WST-1 method), content of malondialdehyde (MDA) (by TBA method), and expression of Golgi matrix protein 130 (GM130), Golgin-84 and Golgi phosphoprotein 3 (GOLPH3) protein and mRNA in lung tissues (by Western blot or real-time polymerase chain reaction). Results:Compared with group C, the lung injury score, W/D ratio, cell apoptosis index, serum IL-6 and TNF-α concentrations and MDA content were significantly increased, SOD activity was decreased, the expression of GM130 and Golgin-84 protein and mRNA was down-regulated, the expression of GOLPH3 protein and mRNA was up-regulated ( P<0.05), and Golgi apparatus was swollen and vacuolated in the other three groups.Compared with group LPS, lung injury score, W/D ratio, cell apoptosis index, serum IL-6 and TNF-α concentrations and MDA content were significantly decreased, SOD activity was increased, the expression of GM130 and Golgin-84 protein and mRNA was up-regulated, the expression of GOLPH3 protein and mRNA was down-regulated ( P<0.05), and swelling and vacuolization of Golgi apparatus were reduced in group EA+ LPS, and no significant change was found in the parameters mentioned above in group SEA+ LPS ( P>0.05). Conclusions:The mechanism by which EA reduces endotoxin-induced ALI may be related to inhibition of Golgi apparatus stress in lung tissues of rats.
7.Practice of relieving emergency overcrowding in Peking Union Medical College Hospital
Jihai LIU ; Xianlin HAN ; Taiping ZHANG ; Quan LIAO ; Xisheng WENG ; Huadong ZHU ; Qing CHANG ; Hui PAN ; Na GUO ; Zhanjie ZHANG ; Yafang LI ; Di SHI ; Fan LI ; Pengxia SUN ; Jun XU ; Wenming WU ; Shuyang ZHANG
Chinese Journal of Hospital Administration 2021;37(6):518-521
Peking Union Medical College Hospital, as one of the most stressful medical institutions in China, is facing the problem of emergency department overcrowding. In order to effectively alleviate the emergency overcrowding, improve the medical quality and patients′ medical experience, the hospital firmly grasped the two incremental links of " throughput" and " output" factors, established a multidisciplinary and multi-department cooperation team, constructed a close medical alliance cooperation mode, and innovated and explored a harmonious emergency overcrowding relief mode with the goal of unblocking the " exit" of patients. The practice showed that the comprehensive measures could effectively alleviate the problem of emergency overcrowding, and improve the medical environment and medical quality.
8.Clinical characteristics of acute myeloid leukemia patients with TET2 gene mutation and effects of TET2 mutation on therapeutic efficacy and prognosis
Wenxia SHI ; Yongxin GUO ; Junjie SHEN ; Wenming CHEN ; Wenwen GUO ; Yanyan TANG ; Shuige YANG ; Zhongliang SUN ; Daoping SUN
Journal of Leukemia & Lymphoma 2021;30(12):719-725
Objective:To investigate clinical features of adult patients with acute myeloid leukemia (AML) with TET2 gene mutation and effects of TET2 mutation on therapeutic efficacy and prognosis.Methods:A total of 123 newly diagnosed adult AML patients (except for acute promyelocytic leukemia) admitted to Jining No.1 People's Hospital from March 2017 to April 2021 were selected. Mutations of 24 AML-related genes including TET2 mutation were detected by using second-generation sequencing technology. Patients were divided into two groups according to the presence of TET2 mutation: TET2 mutation group and TET2 wild type group. The differences in clinicopathological characteristics, short-term efficacy and survival of both groups were compared.Results:Among 123 patients, TET2 mutation was detected in 28 cases (22.8%). Compared with TET2 wild type group, the patients were older [(59±15) years vs.(49±16) years, t = 2.984, P = 0.003], French-American-British (FAB) Corporative Group M 4 and M 5 subtypes were more common [75.0% (21/28) vs. 51.6% (49/95), χ2 = 4.838, P = 0.028], and the positive rate of CD34 in AML patients was lower in TET2 mutation group [46.4% (13/28) vs.72.6% (69/95), χ2 = 6.685, P = 0.010]. Moreover, TET2 mutation was more likely to be accompanied with ZRSR2 mutation [10.7% (3/28) vs. 1.1% (1/95), P = 0.037] and NPM1 mutation [35.7% (10/28) vs.17.9% (17/95), χ2 = 4.008, P = 0.045], but less likely to be accompanied with IDH1/2 mutation [0 vs.17.9% (17/95), P = 0.012]. However, there were no statistically significant differences in gender, peripheral blood leukocyte count at initial diagnosis, hemoglobin level, platelet count, bone marrow blasts ratio, cytogenetics and the European LeukemiaNet (ELN) risk stratification between the two groups (all P>0.05). In addition, there were no significant differences in the overall response rate (ORR) of 1 cycle chemotherapy [75.0% (12/16) vs. 66.7% (42/63), χ2 = 0.410, P = 0.522] and demethylation therapy [66.7% (4/6) vs. 44.4% (8/18), P = 0.640]. The difference in overall survival (OS) of both groups was not statistically significant [median OS time: 23 months (95% CI 5-41 months) vs. 35 months (95% CI 18-52 months, P = 0.498]. Conclusions:In AML patients, TET2 mutation is associated with advanced age, M 4 and M 5 subtypes, and low expression of CD34 on AML blasts. TET2 mutation is commonly accompanied by ZRSR2 and NPM1 mutation, but not IDH1 or IDH2 mutation. TET2 mutation may have no significant effects on therapeutic efficacy and survival in the whole cohort of AML patients without risk stratification.
9.Expert consensus on rehabilitation strategies for traumatic spinal cord injury
Liehu CAO ; Feng NIU ; Wencai ZHANG ; Qiang YANG ; Shijie CHEN ; Guoqing YANG ; Boyu WANG ; Yanxi CHEN ; Guohui LIU ; Dongliang WANG ; Ximing LIU ; Xiaoling TONG ; Guodong LIU ; Hongjian LIU ; Tao LUO ; Zhongmin SHI ; Biaotong HUANG ; Wenming CHEN ; Qining WANG ; Shaojun SONG ; Lili YANG ; Tongsheng LIU ; Dawei HE ; Zhenghong YU ; Jianzheng ZHANG ; Zhiyong HOU ; Zengwu SHAO ; Dianying ZHANG ; Haodong LIN ; Baoqing YU ; Yunfeng CHEN ; Xiaodong ZHU ; Qinglin HANG ; Zhengrong GU ; Xiao CHEN ; Yan HU ; Liming XIONG ; Yunfei ZHANG ; Yong WANG ; Lei ZHANG ; Lei YANG ; Peijian TONG ; Jinpeng JIA ; Peng ZHANG ; Yong ZHANG ; Kuo SUN ; Tao SHEN ; Shiwu DONG ; Jianfei WANG ; Hongliang WANG ; Yong FENG ; Zhimin YING ; Chengdong HU ; Ming LI ; Xiaotao CHEN ; Weiguo YANG ; Xing WU ; Jiaqian ZHOU ; Haidong XU ; Bobin MI ; Yingze ZHANG ; Jiacan SU
Chinese Journal of Trauma 2020;36(5):385-392
TSCI have dyskinesia and sensory disturbance that can cause various life-threaten complications. The patients with traumatic spinal cord injury (TSCI), seriously affecting the quality of life of patients. Based on the epidemiology of TSCI and domestic and foreign literatures as well as expert investigations, this expert consensus reviews the definition, injury classification, rehabilitation assessment, rehabilitation strategies and rehabilitation measures of TSCI so as to provide early standardized rehabilitation treatment methods for TSCI.
10.Population Genetics of SARS-CoV-2:Disentangling Effects of Sampling Bias and Infection Clusters
Liu QI ; Zhao SHILEI ; Shi CHENG-MIN ; Song SHUHUI ; Zhu SIHUI ; Su YANKAI ; Zhao WENMING ; Li MINGKUN ; Bao YIMING ; Xue YONGBIAO ; Chen HUA
Genomics, Proteomics & Bioinformatics 2020;18(6):640-647
A novel RNA virus, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is responsible for the ongoing outbreak of coronavirus disease 2019 (COVID-19). Population genetic analysis could be useful for investigating the origin and evolutionary dynamics of COVID-19. However, due to extensive sampling bias and existence of infection clusters during the epidemic spread, direct applications of existing approaches can lead to biased parameter estima-tions and data misinterpretation. In this study, we first present robust estimator for the time to the most recent common ancestor (TMRCA) and the mutation rate, and then apply the approach to analyze 12,909 genomic sequences of SARS-CoV-2. The mutation rate is inferred to be 8.69 × 10-4 per site per year with a 95% confidence interval (CI) of [8.61 × 10-4, 8.77 × 10-4], and the TMRCA of the samples inferred to be Nov 28, 2019 with a 95% CI of [Oct 20, 2019, Dec 9, 2019]. The results indicate that COVID-19 might originate earlier than and outside of Wuhan Seafood Market. We further demonstrate that genetic polymorphism patterns, including the enrichment of specific haplotypes and the temporal allele frequency trajectories generated from infection clusters, are similar to those caused by evolutionary forces such as natural selection. Our results show that population genetic methods need to be developed to efficiently detangle the effects of sampling bias and infection clusters to gain insights into the evolutionary mechanism ofSARS-CoV-2. Software for implementing VirusMuT can be downloaded at https://bigd.big.ac.cn/biocode/tools/BT007081.


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