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Preparation and verification of severe acute respiratory syndrome Coronavirus 2 Pseudoparticles 严重急性呼吸综合征冠状病毒2型假颗粒的制备与验证
Q4 Medicine Pub Date : 2020-04-01 DOI: 10.16781/J.0258-879X.2020.04.0359
Haoran Peng, Liangliang Jiang, H. Yan-hua, J. Ai, Hailin Tang, Z. Qi, P. Zhao
Objective: To establish a method for preparing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudoparticles (SARS-CoV-2 pps)
目的:建立一种制备严重急性呼吸综合征冠状病毒2 (SARS-CoV-2)假颗粒(SARS-CoV-2 pps)的方法
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引用次数: 0
Severe Coronavirus disease 2019 successfully treated with glucocorticoid and high-dose intravenous immunoglobulin: A case report and analysis of clinical experience 糖皮质激素联合大剂量静脉注射免疫球蛋白治疗2019年重症冠状病毒病成功一例报告及临床经验分析
Q4 Medicine Pub Date : 2020-02-01 DOI: 10.16781/J.0258-879X.2020.02.0181
Kailian Zheng, You-rong Zhang, Qiongya Wang, Ying Xu, Hui Wang, Xiang-Yu Kong, Yi Li, Yuju Dong
Objective To report a case of severe Coronavirus disease 2019 (COVID-19) that had been successfully treated with glucocorticoid and intravenous immunoglobulin therapy. Methods and results The patient was a healthcare provider in Wuhan City who was taking care of COVID-19 patients before the onset of the disease. He started to cough with a little white sticky sputum on January 16, 2020 and had a fever on January 22 (up to 38.5 ℃) before admission. CT results showed mild exudation in both lungs. Oral oseltamivir and intravenous moxifloxacin, cefoperazone and sulbactam sodium were given in addition to nutritional support. On January 26, the patient had chest tightness and shortness of breath. A swab test was positive for severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) nucleic acid, and chest CT results showed moderate exudation in both lungs. On January 28, shortness of breath worsened and intravenous methylprednisolone (40 mg, qd) and immunoglobulin (10 g, qd) were given. On January 30, shortness of breath further worsened;he had a body temperature of 40.7 ℃, pulse oxygen saturation (SpO2) of 83% with oxygen inhalation at 10 L/min, and lymphocyte count of 0.5×109/L. The dose of methylprednisolone and immunoglobulin were adjusted to 40 mg, q12h and 20 g, qd, respectively. Subcutaneous injection of thymalfasin (1.6 mg, qd) was added. Then the body temperature returned to normal, and symptoms such as chest tightness and shortness of breath were gradually improved. On January 31, SpO2 was 88% with oxygen inhalation at 10 L/min and a chest CT results revealed large amount of exudation in both lungs. On February 2, SpO2 was 95% with oxygen inhalation at 5 L/min and the dose of methylprednisolone was then gradually reduced. A chest CT results on February 3 revealed improved lung inflammation, and a throat swab on February 4 and 9 was negative for SARS-CoV-2 nucleic acid. Conclusion Glucocorticoid should be used with caution in patients with early and mild COVID-19. However, appropriate dosage of glucocorticoid can be used to modulate lung inflammation in patients with decompensated respiratory failure. Additionally, large dose of immunoglobulin can be given if necessary.
目的报告1例重症冠状病毒病(COVID-19)经糖皮质激素和静脉注射免疫球蛋白治疗成功的病例。方法与结果患者为武汉市一名医疗服务人员,发病前一直在照顾COVID-19患者。他于2020年1月16日开始咳嗽,痰中有少许白色粘稠,入院前1月22日发烧(高达38.5℃)。CT示双肺轻度渗出。口服奥司他韦,静脉注射莫西沙星、头孢哌酮和舒巴坦钠,并给予营养支持。1月26日,患者出现胸闷、呼吸短促。拭子检测SARS-CoV-2核酸阳性,胸部CT示双肺中度渗出。1月28日,呼吸急促加重,静脉注射甲强的松龙(40 mg, qd)和免疫球蛋白(10 g, qd)。1月30日呼吸急促加重,体温40.7℃,脉搏血氧饱和度(SpO2) 83%,吸氧10 L/min,淋巴细胞计数0.5×109/L。甲基强的松龙和免疫球蛋白的剂量分别调整为40mg, q12h和20g, qd。皮下注射胸腺甲素(1.6 mg, qd)。随后体温恢复正常,胸闷、呼吸短促等症状逐渐好转。1月31日,SpO2 88%,吸氧10l /min,胸部CT示双肺大量渗出。2月2日,以5l /min吸氧,SpO2为95%,随后逐渐减少甲泼尼龙剂量。2月3日胸部CT结果显示肺部炎症好转,2月4日和9日咽拭子检测SARS-CoV-2核酸阴性。结论早期和轻度COVID-19患者应慎用糖皮质激素。然而,适当剂量的糖皮质激素可用于调节失代偿性呼吸衰竭患者的肺部炎症。此外,必要时可给予大剂量免疫球蛋白。
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引用次数: 1
Analysis of amino acid variations of major proteins from severe acute respiratory syndrome coronavirus 2 严重急性呼吸综合征冠状病毒主要蛋白氨基酸变异分析
Q4 Medicine Pub Date : 2020-01-01 DOI: 10.16781/J.0258-879X.2020.05.0465
Q. Zhao-ling, Luo Li
Objective To identify key amino acid variations of major proteins from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by biophysical methods. Methods Through amino acid sequence alignment, classification of variant amino acid residues, three-dimensional structure reconstruction of proteins, and electrostatic interaction analysis of amino acid residues, the key amino acid variations of major proteins from SARS-CoV-2 was analyzed with RaTG13, the bat coronavirus with the highest homology, as the reference. Results At least ten amino acid variations that affect the possible electrostatic interactions were identified in RNA-dependent RNA polymerase (RdRp), exoribonuclease (ExoN), uridylate-specific endoribonuclease (NendoU), and spike (S) protein from SARS-CoV-2. These variations may affect the spatial conformation and biological functions of the proteins. Conclusion The key amino acid variations of the major proteins from SARS-CoV-2 have been preliminarily identified, providing clues for understanding the genetic, pathogenic and epidemiological characteristics of the virus..
目的用生物物理方法鉴定严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)主要蛋白的关键氨基酸变异。方法通过氨基酸序列比对、变异氨基酸残基分类、蛋白质三维结构重建和氨基酸残基静电相互作用分析,以同源性最高的蝙蝠冠状病毒RaTG13为参考,分析SARS-CoV-2主要蛋白的关键氨基酸变异。结果在SARS-CoV-2的RNA依赖性RNA聚合酶(RdRp)、核糖核酸外切酶(ExoN)、尿苷酸特异性核糖核酸内切酶(NendoU)和刺突(S)蛋白中发现了至少10种影响可能的静电相互作用的氨基酸变异。这些变异可能影响蛋白质的空间构象和生物学功能。结论初步鉴定了SARS-CoV-2主要蛋白的关键氨基酸变异,为了解该病毒的遗传、致病和流行病学特征提供了线索。
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引用次数: 0
Prokaryotic expression, purification and antiserum preparation of severe acute respiratory syndrome coronavirus 2 nucleocapsid protein 严重急性呼吸综合征冠状病毒2型核衣壳蛋白的原核表达、纯化及抗血清制备
Q4 Medicine Pub Date : 2020-01-01 DOI: 10.16781/J.0258-879X.2020.05.0481
Qin Zhao-Ling, Peng Haoran, D. Cui-ling, Xiao Ai-Jun, QI Zhong-tian, Z. Ping
Objective To express and purify the recombinant nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and prepare antiserum from immunized mice Methods The prokaryotic plasmid pET28a-N containing SARS-CoV-2 N gene was transformed into Escherichia coli BL21 (DE3) The expression of recombinant SARS-CoV-2 N protein was induced by isopropyl-β-D-thiogalactopyranoside The Ni-NTA affinity chromatography column was used to purify the recombinant SARS-CoV-2 N protein, and antiserum was obtained from the BALB/c mice immunized with recombinant SARS-CoV-2 N protein combined with manganese adjuvant through intramuscular and subcutaneous injections The reactions of recombinant SARS-CoV-2 N protein with SARS-CoV-2 N monoclonal antibodies and severe acute respiratory syndrome coronavirus (SARS-CoV) N polyclonal antibodies were detected by Western blotting The reaction of mouse antiserum with the recombinant SARS-CoV-2 N protein expressed in the cells transfected with eukaryotic expression plasmid was examined by indirect immunofluorescence assay Results The recombinant SARS-CoV-2 N protein was successfully induced and expressed as a soluble protein with a molecular weight of about 55 000 High concentration of purified protein was obtained The results of Western blotting showed that the recombinant SARS-CoV-2 N protein could be specifically recognized by the SARS-CoV-2 N monoclonal antibodies and the SARS-CoV N polyclonal antibodies The prepared mouse antiserum could also correctly recognize the recombinant SARS-CoV-2 N protein expressed in mammalian cells by indirect immunofluorescence assay Conclusion Recombinant SARS-CoV-2 N protein has been successfully expressed and purified from the prokaryotic expression system, and mouse antiserum has been prepared, which lays a foundation for establishing a rapid SARS-CoV-2 diagnostic tool and further studying the function of SARS-CoV-2 N protein
目的表达和纯化重组SARS-CoV-2病毒核衣壳(N)蛋白,并从免疫小鼠制备抗血清。方法将含有SARS-CoV-2 N基因的原核质粒pET28a-N转化至大肠杆菌BL21 (DE3),用异丙基-β- d -硫代半乙酰核苷诱导表达重组SARS-CoV-2 N蛋白,采用Ni-NTA亲和层析柱纯化重组SARS-CoV-2 N蛋白。通过肌肉和皮下注射重组SARS-CoV- 2n蛋白联合锰佐剂免疫BALB/c小鼠,获得重组SARS-CoV- 2n蛋白与SARS-CoV- 2n单克隆抗体和严重急性呼吸综合征冠状病毒(SARS-CoV) N多克隆抗体的反应,采用Western blotting检测小鼠抗血清与转染细胞中表达的重组SARS-CoV- 2n蛋白的反应结果成功诱导重组SARS-CoV- 2n蛋白,并表达为分子量约为55000的可溶性蛋白,获得了高浓度的纯化蛋白。Western blotting结果表明,重组SARS-CoV- 2n蛋白可被制备的小鼠SARS-CoV- 2n单克隆抗体和SARS-CoV- 2n多克隆抗体特异性识别结论在原核表达系统中成功表达并纯化了重组sars - cov - 2n蛋白,制备了小鼠抗血清,为建立快速诊断工具和进一步研究sars - cov - 2n蛋白的功能奠定了基础
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引用次数: 1
Application of bronchoscopy in diagnosis of coronavirus disease 2019 and related occupation protection 支气管镜在2019冠状病毒病诊断及相关职业防护中的应用
Q4 Medicine Pub Date : 2020-01-01 DOI: 10.16781/J.0258-879X.2020.05.0498
Ning Yunye, Huang Haidong, Bai Chong
Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARSCoV- 2) has become a global pandemic and led to a serious impact on public health and economic development. This paper summarizes the source of the first isolated and identified SARS-CoV-2 samples and the problems present in the etiology detection of virus RNA. The necessity and limitation of bronchoscopy use in the diagnosis of COVID-19 were discussed and the occupation protection measures for bronchoscopy in COVID-19 were emphasized. It is important to accelerate the development of new disposable protective devices for the bronchoscopic examination..
由严重急性呼吸综合征冠状病毒2型(SARSCoV- 2)引起的冠状病毒病2019 (COVID-19)已成为全球性流行病,对公共卫生和经济发展造成严重影响。本文综述了首次分离鉴定SARS-CoV-2样本的来源及病毒RNA病原学检测中存在的问题。讨论了支气管镜检查在新冠肺炎诊断中的必要性和局限性,强调了新冠肺炎患者支气管镜检查的职业防护措施。因此,加快开发新型一次性支气管镜检查用保护装置是十分重要的。
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引用次数: 0
Computed tomography in coronavirus disease 2019: diagnosis and clinical significance 2019冠状病毒病的ct诊断及临床意义
Q4 Medicine Pub Date : 2020-01-01 DOI: 10.16781/J.0258-879X.2020.06.0588
Fang Xu, Bian Yun, L. Fang, J. Tao, Shao Cheng-wei, Wang Li, L. Jian-ping
Objective To investigate the computed tomography (CT) features of the coronavirus disease 2019 (COVID-19) and the clinical significance, so as to improve our understanding of CT imaging of this disease. Methods The chest CT features of seven COVID-19 patients, who were diagnosed by virus nucleic acid test from Jan. 25 to Feb. 15, 2020 in Changhai Hospital of Naval Medical University (Second Military Medical University), were analyzed retrospectively. There were six males and one female, aged (51.1±18.8) years (range 29-75 years). All the seven patients received chest CT plain scan examimation. The CT images were interpreted by two experienced senior radiologists, and the distribution, location and density of lesions, number of involved lobes, air bronchogram, mediastinal lymphadenopathy and pleural effusion were analyzed. Results The average time from onset of symptoms to CT examination was 3.6 d (range 1-9 d) in the seven COVID-19 patients. The lesions were distributed in single lung in one case and bilateral lungs in six cases. The lesions involved middle and lateral fields of lungs in five cases and the whole field of lungs in two cases. The lesions showed ground-glass opacity in four cases and mixed shadow in three cases. The lesions involved two or less lobes in four cases and five lobes in three cases. One case had air bronchogram. No mediastinal lymphadenopathy or pleural effusion were found. Conclusion COVID-19 patients have characteristic CT findings, which has important clinical significance for the diagnosis and treatment of COVID-19. However, the diagnosis should be confirmed based on the patient's epidemic history, clinical symptoms and laboratory indicators.
目的探讨新型冠状病毒病2019 (COVID-19)的CT表现特征及临床意义,提高对该疾病的CT影像学认识。方法回顾性分析海军医科大学(第二军医大学)长海医院2020年1月25日至2月15日经病毒核酸检测确诊的7例新冠肺炎患者的胸部CT表现。男性6例,女性1例,年龄(51.1±18.8)岁(29 ~ 75岁)。7例患者均行胸部CT平扫检查。由两名经验丰富的资深放射科医师对CT图像进行解读,分析病变的分布、位置、密度、受累肺叶数、支气管空气征、纵隔淋巴结病变及胸腔积液。结果7例新冠肺炎患者从出现症状到CT检查的平均时间为3.6 d(范围1 ~ 9 d)。病变分布于单肺1例,双肺6例。病变累及肺中外侧野5例,累及全肺野2例。病变表现为磨玻璃影4例,混影3例。病变涉及两个或更少的叶在四个病例和五个叶在三个病例。1例有支气管充气征。未见纵隔淋巴结病或胸腔积液。结论新冠肺炎患者具有特征性的CT表现,对新冠肺炎的诊治具有重要的临床意义。但应根据患者的流行史、临床症状和实验室指标进行确诊。
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引用次数: 1
Clinical features and imaging findings in six coronavirus disease 2019 patients 6例2019冠状病毒病患者的临床特征和影像学表现
Q4 Medicine Pub Date : 2020-01-01 DOI: 10.16781/J.0258-879X.2020.06.0616
Ge Xia-hui, X. Jing, Ha Ming-hao, Zhang Li-wei, Song Li-tao, Wang Jie-ning, Lin Yan, G. Xiao-yan, Bai Chong
Objective To summarize the clinical features and imaging findings of six coronavirus disease 2019 (COVID-19) patients, so as to provide evidences for early diagnosis and clinical intervention. Methods Six COVID-19 patients with positive severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were enrolled from the Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine from Jan. 1 to Feb. 22, 2020. The epidemiological history, clinical manifestations, imaging data and laboratory indicators were retrospectively analyzed. Results All six patients had a clear travel or residence history in Wuhan. Four patients had fever, three had cough, two had upper respiratory tract symptoms such as runny nose and sore throat, and two had systemic symptoms such as headache and muscle ache. Chest computed tomography (CT) showed that all the six patients had abnormal manifestations in bilateral lungs, and the lower lung lesions were more common than the upper lung lesions. The main manifestations were multiple ground-glass opacities, consolidation shadows, crazy paving sign and different degrees of fibrosis in lateral field of bilateral lungs. Chest CT examination later after onset showed lung consolidation and severe fibrosis. Conclusion The imaging of COVID-19 has special characteristics. Combined with the epidemiological history, clinical manifestations and the detection of SARS-CoV-2 nucleic acid, COVID-19 can be effectively diagnosed in the early stage.
目的总结6例新型冠状病毒病(COVID-19)患者的临床特点及影像学表现,为早期诊断和临床干预提供依据。方法选取2020年1月1日至2月22日上海中医药大学第七人民医院收治的6例新型冠状病毒感染(SARS-CoV-2)阳性患者。回顾性分析流行病学病史、临床表现、影像学资料及实验室指标。结果6例患者均有明确的武汉市旅行或居住史。发热4例,咳嗽3例,流鼻水、喉咙痛等上呼吸道症状2例,头痛、肌肉酸痛等全身症状2例。胸部CT显示6例患者均有双肺异常表现,且下肺病变多于上肺病变。主要表现为双侧肺侧野多发磨玻璃影、实变影、疯狂铺路征及不同程度纤维化。发病后胸部CT检查显示肺实变及严重纤维化。结论新型冠状病毒肺炎影像学表现具有特殊性。结合流行病学史、临床表现和SARS-CoV-2核酸检测,可早期有效诊断COVID-19。
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引用次数: 0
Traditional Chinese medicine syndrome differentiation of coronavirus disease 2019 patients 2019冠状病毒病患者的中医辨证
Q4 Medicine Pub Date : 2020-01-01 DOI: 10.16781/J.0258-879X.2020.05.0493
X. Meng, X. Wan, Jun Yu Li, Xiao-Li Gong, Yujiao Liang, Song Gao, Ji Ping Xu, Luyang Li, Xiao-Qiang Yue
Objective To explore the rule of traditional Chinese medicine (TCM) syndrome differentiation of coronavirus disease 2019 (COVID-19) patients. Methods The symptoms of 756 cases with COVID-19 in Guanggu Branch of Maternity and Child Healthcare Hospital of Hubei Province were collected by cross sectional survey. The incidence rates of the symptoms were recorded by frequency method at different courses of the disease: prodromal stage (onset), middle stage (7-30 days), and later stage (>30 days). The common symptoms (incidence rate>5.0%) were analyzed by systematic clustering. With expert experience, the rule of TCM syndrome differentiation of COVID-19 patients was summarized. Results Fever (52.25%, 395 cases), cough (43.25%, 327 cases), asthenia (27.25%, 206 cases), chest distress (26.72%, 202 cases), asthma (17.59%, 133 cases) and expectoration (5.03%, 38 cases) were the most common symptoms in the prodromal stage (756 cases) of the disease, which were clustered into one category except expectoration, indicating the pathogenesis of both lung and body surface suppressed by dampness. In the middle stage (383 cases), the 19 common symptoms including greasy fur (64.49%, 247 cases), yellow fur (43.86%, 168 cases), thick fur (40.21%, 154 cases), cough (34.73%, 133 cases), red tongue (32.38%, 124 cases), poor stool (25.85%, 99 cases), asthma (25.33%, 97 cases), asthenia (25.07%, 96 cases), poor appetite (23.76%, 91 cases), bitterness of mouth (14.36%, 55 cases), dry fur (12.01%, 46 cases), purple tongue (12.01%, 46 cases), perspiration (11.49%, 44 cases), constipation (10.18%, 39 cases), white phlegm (8.62%, 33 cases), insomnia (7.31%, 28 cases), nausea (7.05%, 27 cases), diarrhea (6.79%, 26 cases) and yellow phlegm (6.27%, 24 cases), were clustered into three groups, indicating the pathogenesis of damp-heat accumulation, obstruction of lung and spleen by dampness, and dryness due to dampness-heat. In the later stage (373 cases), the 13 common symptoms including greasy fur (50.94%, 190 cases), asthenia (39.41%, 147 cases), cough (37.80%, 141 cases), red tongue (33.78%, 126 cases), asthma (32.17%, 120 cases), perspiration (23.86%, 89 cases), dry mouth (22.79%, 85 cases), poor appetite (20.11%, 75 cases), poor stool (19.30%, 72 cases), bitterness of mouth (15.01%, 56 cases), white phlegm (10.72%, 40 cases), palpitation (8.31%, 31 cases) and little fur (8.04%, 30 cases), were clustered into two groups, indicating the pathogenesis of deficiency of Qi and Yin with residual dampness, and deficiency of lung Qi and spleen Qi with residual dampness. Conclusion The TCM syndromes of COVID-19 patients in different stages have its own typical characteristics, with a regular change from exterior to interior, from dampness to dampness-heat and from excess to deficiency..
目的探讨新型冠状病毒病(COVID-19)患者的中医辨证规律。方法采用横断面调查法收集湖北省广谷妇幼保健院756例新冠肺炎患者的症状。采用频率法记录症状在不同病程的发生率:前驱期(发病)、中期(7 ~ 30 d)、晚期(10 ~ 30 d)。采用系统聚类分析常见症状(发病率>5.0%)。结合专家经验,总结新冠肺炎患者中医辨证规律。结果本病前驱期(756例)以发热(52.25%,395例)、咳嗽(43.25%,327例)、乏力(27.25%,206例)、胸闷(26.72%,202例)、哮喘(17.59%,133例)、咳痰(5.03%,38例)为最常见症状,除咳痰外均聚为一类,提示肺、体表湿抑均为发病机制。中期(383例),19种常见症状包括毛油腻(64.49%,247例)、毛黄(43.86%,168例)、毛厚(40.21%,154例)、咳嗽(34.73%,133例)、舌红(32.38%,124例)、大便不畅(25.85%,99例)、哮喘(25.33%,97例)、乏力(25.07%,96例)、食欲不佳(23.76%,91例)、口苦(14.36%,55例)、毛干(12.01%,46例)、舌紫(12.01%,46例)、出汗(11.49%,44例)、便秘(10.18%,168例)。其中痰白(8.62%,33例)、失眠(7.31%,28例)、恶心(7.05%,27例)、腹泻(6.79%,26例)、黄痰(6.27%,24例)为3组,提示病机以湿热积存、湿阻肺脾、湿热燥为主。晚期(373例)13种常见症状为毛油腻(50.94%,190例)、乏力(39.41%,147例)、咳嗽(37.80%,141例)、舌红(33.78%,126例)、哮喘(32.17%,120例)、出汗(23.86%,89例)、口干(22.79%,85例)、食欲不振(20.11%,75例)、大便不畅(19.30%,72例)、口苦(15.01%,56例)、痰白(10.72%,40例)、心悸(8.31%,31例)、毛少(8.04%,30例)。分为两组,提示气阴虚湿型病机和肺气脾虚湿型病机。结论新冠肺炎不同分期中医证候具有自身的典型特征,由外证到内证、由湿证到湿热证、由虚证到虚证的变化规律明显。
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引用次数: 0
Application of cell lineage reprogramming for central nervous system injury and repair 细胞谱系重编程在中枢神经系统损伤与修复中的应用
Q4 Medicine Pub Date : 2016-06-01 DOI: 10.16781/J.0258-879X.2016.06.0729
Xiu Sun, Yi-min Yuan, Xin Hu, Shangyao Qin, Z. Su
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引用次数: 0
A comparative study of nanoknife, cryoablation, and radiofrequency ablation for treatment of pancreatic cancer xenograft in rabbits 纳米刀、冷冻消融术和射频消融术治疗兔胰腺癌异种移植的比较研究
Q4 Medicine Pub Date : 2016-01-01 DOI: 10.16781/J.0258-879X.2016.01.0010
Z. Ning, Peng Wang, Hao Chen, Jing Xie, Jun-hua Lin, Xiao-yan Zhu, Qi Chen, L. Xu, Li-bin Song, Song Gao, Feng Jiang, Z. Meng
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引用次数: 0
期刊
第二军医大学学报
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