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Cerebrospinal fluid used as culture medium prior to autologous olfactory ensheathing cell transplantation
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作者 Weijiang Wu Qing Lan +4 位作者 Hua Lu Aihua Zhu Yunzhao Jiang Ge Chen Guozhen Hui 《Neural Regeneration Research》 SCIE CAS CSCD 2010年第1期21-27,共7页
BACKGROUND: Cerebrospinal fluid can be an inducer for neural stem cells in vitro, but few studies employ cerebrospinal fluid to culture olfactory ensheathing cells. OBJECTIVE: To investigate the growth of nasal muco... BACKGROUND: Cerebrospinal fluid can be an inducer for neural stem cells in vitro, but few studies employ cerebrospinal fluid to culture olfactory ensheathing cells. OBJECTIVE: To investigate the growth of nasal mucosa olfactory ensheathing cells in normal cerebrospinal fluid, and to analyze the feasibility of cerebrospinal fluid for culturing olfactory ensheathing cells used for transplantation. DESIGN, TIME AND SETTING: A completely randomized, block design study was performed at the Cell Laboratory, Wuxi Third People's Hospital, and Jiangsu Institute of Parasitic Diseases, China, in August 2008. MATERIALS: Dulbecco's modified Eagle's medium/F12 (DMEM/F12) and fetal bovine serum (Gibco BRL, USA), mouse anti-rat P75 monoclonal antibody and rabbit anti-glial fibrillary acidic protein polyclonal anti body ('Santa Cruz Biotechnology, USA), mouse anti-rat myelin basic protein monoclonal antibody (Cymbus, UK), mouse anti-rat microtubule-associated protein-2 monoclonal antibody (Transduction Laboratories, USA), FITC conjugated rabbit anti-mouse monoclonal antibody (Boster, China), TRITC conjugated goat anti-rabbit monoclonal antibody (Sigma, USA) were used. METHODS: Nasal mucosa olfactory ensheathing cells were separately incubated in DMEM/F12, cerebrospinal fluid, and changing DMEM/F12 into cerebrospinal fluid. Adult female Sprague Dawley rat models of spinal hemisection were established. Nerve injury was repaired by transplantation of nasal mucosa olfactory ensheathing cells cultured in cerebrospinal fluid or DMEM/F12. MAIN OUTCOME MEASURES: The proliferative ability of olfactory ensheathing cells cultured in cerebrospinal fluid was determined by a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. The morphology and purity of olfactory ensheathing cells were detected using immunohistochemistry. Animal behavior was evaluated by the Basso, Beattie and Bresnahan locomotor rating scale. Morphological repair was assessed by a horseradish peroxidase-tetramethylbenzidine retrograde tracer technique and immunohistochemistry. RESULTS: Changing from DMEM/F12 to cerebrospinal fluid did not change overall culture morphology and purity on day 14. These cells also contributed to myelinization and the conduction velocity of regenerated axons, and improved motor abilities of denervated muscle fibers in rats with spinal cord injury. The recovery of behavioral function and neuronal regeneration was similar in the two groups. CONCLUSION: Cerebrospinal fluid culture prior to autologous olfactory ensheathing cell transplantation is feasible for clinical use. 展开更多
关键词 cerebrospinal fluid olfactory ensheathing cells in vitro culture cell transplantation
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Association of the microbial culture of follicular fluid,vaginal swab and catheter tip with β-hCG IVF positive and negative
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作者 Manoj Chellani Manju Chellani Sandeep Rahangdale 《Asian pacific Journal of Reproduction》 2022年第5期217-222,共6页
Objective:To find out the association of microbial contamination withβ-human chorionic gonadotropin(β-hCG)in-vitro fertilization(IVF)positive and negative.Methods:A total of 73 fresh IVF cycle women were included in... Objective:To find out the association of microbial contamination withβ-human chorionic gonadotropin(β-hCG)in-vitro fertilization(IVF)positive and negative.Methods:A total of 73 fresh IVF cycle women were included in the retrospective study.Vaginal swab culture samples were collected prior to ovum pick-up and embryo transfer.The follicular fluids were collected during ovum pick-up and catheter tip culture samples were collected after successful embryo transfer.After 14 days of the embryo transfer,women were classified intoβ-hCG IVF positive and negative.The comparative statistical analyses of aerobic microbial culture reports were done betweenβ-hCG IVF positive and negative women.Results:Out of 73 women,42(57.5%)were found to beβ-hCG IVF positive and 31(43.5%)were negative.In the aerobic culture of ovum pick-up vaginal swab,follicular fluid,embryo transplantation vaginal swab and catheter tip,Enterococcus faecalis was found to be higher compared to other bacteria(Streptococcus spp.,Candida,Escherichia coli and Klebsiella).Regarding the comparison between IVF positive and negative,the overall microbial infection rate of vaginal swab culture during ovum pick-up and embryo transplantation was found to be higher in IVF negative women than in IVF positive women(38.71%vs.28.57%);however,it was not statistically significant(P>0.05).The follicular fluid and catheter tip culture microbial infection rate was found to slightly higher in IVF positive women than in IVF negative women(54.76%vs.41.94%and 19.05%vs.9.68%,respectively),but there were not significant differences(P>0.05).Conclusions:The aerobic microbial culture reports of follicular fluid,vaginal swab culture,and catheter tip culture are not statistically significantly withβ-hCG IVF positive. 展开更多
关键词 Follicular fluid culture Vaginal swab culture Catheter tip culture β-human chorionic gonadotropin In-vitro fertilization outcome
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Coscinium fenestratum: Callus and Suspension Cell Culture of the Endangered Medicinal Plant Using Vermicompost Extract and Coelomic Fluid as Plant Tissue Culture Media
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作者 Suman Kashyap Neera Kapoor Radha D. Kale 《American Journal of Plant Sciences》 2016年第6期899-906,共8页
In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were in... In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were induced to form callus when cultured on vermicompost extract media along with coelomic fluid. Suspension medium was developed using vermicompost extract and coelomic fluid in 3:1 ratio. Phytochemical analysis of the alkaloid berberine was confirmed from callus, suspension cell culture and suspension medium by Thin Layer Chromatography and High Performance Liquid Chromatography. Vermicompost and its extracts with coelomic fluid have shown maximum (100 per cent) response of callus induction. Callus mass enlarged with increasing concentration of coelomic fluid and callus growth was assessed from the biomass. Incubation of culture tubes in dark supported callus development significantly. The Rf value of 0.36 confirmed the presence of berberine by Thin Layer Chromatography. Qualitative analysis confirmed the presence of alkaloid berberine with the retention time of 2.8 minutes similar to that of standard reference sample from Sigma chemicals, USA. The suspension medium turned deep yellow because of the release of the alkaloid. Vermicompost and its extracts along with coelomic fluid have shown the economical approach for micropropagation of economically and medicinally important plants. 展开更多
关键词 BERBERINE Coelomic fluid Coscinium fenestratum Plant Tissue culture Media Suspension Cell culture Vermicompost Extract
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MORPHOLOGICAL MODEL FOR RHEOLOGICAL PROPERTIES OF PLANT CELL SUSPENSION CULTURE SYSTEM
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作者 吴兆亮 元英进 +1 位作者 邢允 胡宗定 《Transactions of Tianjin University》 EI CAS 1999年第2期61-64,共4页
This paper puts forward a physical and mathematical model for the rheological properties of a plant cell suspension culture system.The model can explain why the system is pseudoplastic satisfactorily,thus the rheologi... This paper puts forward a physical and mathematical model for the rheological properties of a plant cell suspension culture system.The model can explain why the system is pseudoplastic satisfactorily,thus the rheological properties of the system as the effect of the flow behavior index on plant cell concentration are interpreted correctly and the mechanism of the rheological properties of the system is further understood.Therefore the model can be applied in the technological design and optimum conditions of the system and the reformation,evaluation and scale up of reactors. 展开更多
关键词 morphological model rheological property plant cell suspension culture pseudoplastic fluid
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Mean platelet volume as a novel predictor of systematic inflammatory response in cirrhotic patients with culturenegative neutrocytic ascites 被引量:2
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作者 Marisol Galvez-Martinez Alfredo I Servin-Caamano +3 位作者 Eduardo Perez-Torres Francisco Salas-Gordillo Xaira Rivera-Gutierrez Fatima Higuera-de la Tijera 《World Journal of Hepatology》 CAS 2015年第7期1001-1006,共6页
AIM: To identify a mean platelet volume(MPV) cutoff value which should be able to predict the presence of bacterial infection.METHODS: An observational, analytic, retrospective study. We evaluated medical records of c... AIM: To identify a mean platelet volume(MPV) cutoff value which should be able to predict the presence of bacterial infection.METHODS: An observational, analytic, retrospective study. We evaluated medical records of cirrhotic patients who were hospitalized from January 2012 to January 2014 at the Gastroenterology Department of "Hospital General de México Dr. Eduardo Liceaga", we included 51 cirrhotic patients with ascites fluid infection(AFI), and 50 non-infected cirrhotic patients as control group. Receiver operator characteristic curves were used to identify the best cutoff value of several parameters from hematic cytometry, including MPV, to predict the presence of ascites fluid infection.RESULTS: Of the 51 cases with AFI, 48 patients(94.1%) had culture-negative neutrocytic ascites(CNNA), 2(3.9%) had bacterial ascites, and one(2%)had spontaneous bacterial peritonitis. Infected patients had greater count of leucocytes and polymorphonuclear cells, greater levels of MPV and cardiac frequency(P < 0.0001), and lower mean arterial pressure compared with non-infected patients(P = 0.009). Leucocytes, polymorphonuclear count, MPV and cardiac frequency resulted to be good or very good predictive variables of presence of AFI in cirrhotic patients(area under the receiving operating characteristic > 0.80). A cutoff MPV value of 8.3 fl was the best to discriminate between cirrhotic patients with AFI and those without infection. CONCLUSION: Our results support that MPV can be an useful predictor of systemic inflammatory response syndrome in cirrhotic patients with AFI, particularly CNNA. 展开更多
关键词 Mean platelet volume CIRRHOSIS Ascites fluid infection culture negative neutrocytic ascites Systemic inflammatory response
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NEUROBIOLOGICAL EFFECTS OF NRCF DERIVED FROM DISTAL STUMPS OF MOTOR NERVE AND SENSORY NERVE AND b-FGF ON CULTURED MOTONEURON IN VITRO 被引量:1
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作者 张涤生 干季良 +1 位作者 李青峰 平萍 《Journal of Shanghai Second Medical University(Foreign Language Edition)》 2001年第2期91-95,共5页
Objective: To explore the mechanism of neurotropism in peripheral nerve regeneration by assessing the bioactivity of regeneration microcircumstance on motoneurons. Methods The motor branch Of femoral nerve to quadrice... Objective: To explore the mechanism of neurotropism in peripheral nerve regeneration by assessing the bioactivity of regeneration microcircumstance on motoneurons. Methods The motor branch Of femoral nerve to quadriceps was incised and the distal stump was sutured with one-end silicone chamber. The nerve regeneration chamber fluids from distal motor nerve stumps (motor branch of femoral nerve ) (MD-NRCF) was collected 7d post-operatively, and with the same method, nerve regeneration conditioned fluids from distal stumps nerve stumps (saphenous nerve ) (SD-NRCF) was collected. The dissociated rat’s motoneurons were co-cultured with MD-NRCF, SD-NRCF, basic fibroblast growth factor (b-FGF) and serum-free medium for 72h respectively and then were photographed under phase-contrast microscope. The longest neurites and cellbody areas of motoneurons from each group were measured by cell image processing computer system. MTT colorimetric assay was also used to measure cell activation. Results The cells of MD-NRCF group had significantly longer neurites than the other 3 groups, and their activation was also superior to those of the other groups. Conclusion These results indicate that MD-NRCF has more significant neurite-promoting and neurobiological effects on motoneuron than SD-NRCF and b-FGF. 展开更多
关键词 neurotropism nerve regeneration conditions fluid nerve regeneration cultured motoneuron
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蔬菜膳食纤维基多菌群乳酸菌生物膜的制备、表征及其稳定性分析
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作者 吕嘉枥 杜倩茹 +1 位作者 杨柳青 徐颖 《陕西科技大学学报》 北大核心 2024年第1期28-36,共9页
试验采用自主设计的生物膜形成培养装置,以8种乳酸菌类益生菌为菌种,以MRS复合甘蓝、芹菜、黄瓜、冬瓜等4种蔬菜浆液为营养液,以营养液中的水不溶性蔬菜膳食纤维为载体,进行连续动态培养.结果表明,37℃培养7天时,形成的乳酸菌与蔬菜膳... 试验采用自主设计的生物膜形成培养装置,以8种乳酸菌类益生菌为菌种,以MRS复合甘蓝、芹菜、黄瓜、冬瓜等4种蔬菜浆液为营养液,以营养液中的水不溶性蔬菜膳食纤维为载体,进行连续动态培养.结果表明,37℃培养7天时,形成的乳酸菌与蔬菜膳食纤维复合生物膜中的菌浓可达2.24×10^(12)CFU/g,显微表征和菌群结构分析结果表明,乳杆菌属和片球菌属均得到了富集.对获得的乳酸菌与蔬菜膳食纤维复合生物膜进行了贮存试验及模拟消化试验,结果显示25℃6个月后,活菌数仍在10^(6)CFU/g以上,模拟消化后活菌数保持在10^(12)CFU/g以上.本试验结果可为高菌浓益生菌发酵剂、益生菌包埋材料以及益生菌膳食补充剂等的开发提供理论依据及技术支撑. 展开更多
关键词 乳酸菌 蔬菜膳食纤维 生物膜 培养装置 高通量测序 消化液耐受性
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面向空间细胞培养用多路切换陶瓷阀仿真优化研究
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作者 郭淼 谭映军 +7 位作者 陈博博 施镠佳 李芳 王春艳 李越 聂捷琳 刘仲馨 李莹辉 《载人航天》 CSCD 北大核心 2024年第4期526-535,共10页
为了满足空间站环境细胞长期自动培养的需求,研制一款新型细胞自动培养装置,装置由微量泵、多路切换阀等组成。通过设计一款多路切换陶瓷阀,实现仅由一组泵阀自由切换多条液路。通过有限元仿真分析培养液、消化液和裂解液3种液体在多路... 为了满足空间站环境细胞长期自动培养的需求,研制一款新型细胞自动培养装置,装置由微量泵、多路切换阀等组成。通过设计一款多路切换陶瓷阀,实现仅由一组泵阀自由切换多条液路。通过有限元仿真分析培养液、消化液和裂解液3种液体在多路切换阀内的压力、温度、流速等物理参数分布情况,确定最佳阀位布局,并优化阀体结构,以改善不同液体流入阀腔后分布均匀性。优化后多路切换阀内细胞培养液和消化液的压力和温度分布波动率均小于1%,流速稳定。裂解液温度分布变化在优化前为2℃,优化后小于0.01℃。液体流动更加顺畅,滞留现象减少。仿真结果表明:优化后的陶瓷阀有助于提升空间细胞培养的稳定性和经济性。 展开更多
关键词 空间细胞培养 多路切换阀 计算流体力学 有限元分析 陶瓷阀
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传代再培养在孕中晚期孕妇羊水穿刺产前诊断中的应用价值
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作者 景亚玲 唐丽萍 +2 位作者 赵连芳 赵咏梅 张传英 《当代医学》 2024年第5期155-158,共4页
目的探讨传代再培养在孕中、晚期孕妇羊水穿刺产前诊断中的应用价值。方法选取2019年6月至2021年6月于遂宁市中心医院产前接受羊水穿刺并经胎儿染色体核型分析的134例孕中、晚期孕妇羊水标本,每名孕妇羊水标本分为2份,分别采用羊水原瓶... 目的探讨传代再培养在孕中、晚期孕妇羊水穿刺产前诊断中的应用价值。方法选取2019年6月至2021年6月于遂宁市中心医院产前接受羊水穿刺并经胎儿染色体核型分析的134例孕中、晚期孕妇羊水标本,每名孕妇羊水标本分为2份,分别采用羊水原瓶培养和羊水传代再培养方式。分析孕妇孕龄及羊水穿刺适应证分布情况,比较不同培养方式的羊水培养结果(羊水细胞培养成功率、总培养时间)、核型制备质量(细胞克隆数、可制片数、可分析分裂相数),分析羊水核型结果和妊娠结局。结果134例孕中、晚期羊水穿刺孕妇中,穿刺时间以孕23~24^(+6)周(47.01%)为主,超声异常(63.43%)为羊水穿刺最常见适应证。羊水传代再培养方式羊水细胞培养成功率为100.00%,高于羊水原瓶培养方式的70.90%,总培养时间长于羊水原瓶培养方式,细胞克隆数、可制片数和可分析分裂相数均多于羊水原瓶培养方式,差异有统计学意义(P<0.05)。134例孕妇羊水核型中,正常核型120例,异常核型8例,染色体异常检出率为5.97%,染色体多态性6例;除3例47,XN,+21和2例47,XXY孕妇终止妊娠,其他孕妇均继续妊娠。结论孕中、晚期羊水传代再培养方式可提高羊水细胞培养成功率和核型制备质量,对预防出生缺陷具有重要意义。 展开更多
关键词 孕中、晚期 产前诊断 核型分析 羊水培养
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无创胚胎植入前遗传学检测技术进展研究
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作者 王嘉祥 田甜 +2 位作者 张楠 韩子川 杨蕊 《生殖医学杂志》 CAS 2024年第7期965-970,共6页
胚胎植入前遗传学检测(PGT)是辅助生殖与遗传学检测技术相结合的一种胚胎诊断检测技术,在移植前筛查胚胎染色体以排除染色体异常的胚胎。但是由于目前该技术复杂昂贵,且有创操作可能存在安全风险,所以该技术在临床的应用受到一定限制。... 胚胎植入前遗传学检测(PGT)是辅助生殖与遗传学检测技术相结合的一种胚胎诊断检测技术,在移植前筛查胚胎染色体以排除染色体异常的胚胎。但是由于目前该技术复杂昂贵,且有创操作可能存在安全风险,所以该技术在临床的应用受到一定限制。因此,人们开始探索利用囊胚腔液和胚胎培养液中发现的游离DNA进行无创PGT。作为更加安全和便捷的基因检测技术,目前无创PGT已经在胚胎染色体检测等方面取得了一定进展,但现有的研究结果之间差异较大,临床应用的有效性仍存在争议。本文就游离DNA的来源及产生机制、无创PGT的临床研究现状进行综述,为无创PGT应用于临床实践提供参考。 展开更多
关键词 无创胚胎植入前遗传学检测 游离DNA 胚胎培养液 囊胚腔液 遗传污染
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Eikenella corrodens isolated from pleural effusion:A case report
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作者 Zi-Cong Liang Hao Ouyang +5 位作者 Xue-Jing Song Jian-Xin Liang Wei-Hua Zheng Jian-Jun Chen Zheng-Gang Yin Sha-Yan Chen 《World Journal of Clinical Cases》 SCIE 2024年第18期3596-3602,共7页
BACKGROUND The bacterium Eikenella,classified as a gram-negative member of the phylum Proteobacteria,is distinguished by its rarity,corrosive nature,facultative anaerobic properties,and conditional pathogenicity.It re... BACKGROUND The bacterium Eikenella,classified as a gram-negative member of the phylum Proteobacteria,is distinguished by its rarity,corrosive nature,facultative anaerobic properties,and conditional pathogenicity.It represents the sole species within its genus-Eikenella corrodens(E.corrodens)-and can be found colonizing both human and animal oral and nasopharyngeal regions.Additionally,it occasionally inhabits the gastrointestinal or urogenital tracts.However,its slow growth rate can be attributed to its high nutritional requirements.However,there is an uneven distribution of construction and diagnostic capacity in China which poses undeniable challenges for the clinical examination and analysis of this case,especially in the basic hospitals.CASE SUMMARY Here we presented a case of empyema associated with E.corrodens infection in a 67-year-old male patient without any previous history of infectious diseases in our primary hospital in Dongguan district of China.The patient was admitted due to recurrent worsening cough,sputum production,and dyspnea for 3 d,which had persisted for over 20 years.Moreover,the patient experienced a onehour episode of unconsciousness.Upon admission,immediate comprehensive examinations were conducted on the patient which subsequently led to his admission to the intensive care unit.Meanwhile,the patient presented with drowsiness and profuse sweating along with bilateral conjunctival edema observed during initiation of non-invasive ventilation,suggesting empyema.A significant amount of coffee-colored malodorous pleural fluid was drained during the procedure above and sent to the laboratory department for inspection.Finally,laboratory culture results confirmed the presence of E.corrodens infection in the pleural fluid sample.The patient received antimicrobial therapy until died on day 22 in the hospital.CONCLUSION In this report,we presented a case of empyema associated with E.corrodens infection.Multiple courses of morphological examination,viable culture analysis,and biochemical identification revealed its difficulties in detecting distinctive characteristics,as well as a detection model worth promoting.It’s just that there were still certain deficiencies in terms of morphological assessment,biochemical identification,and drug susceptibility testing. 展开更多
关键词 Eikenella corrodens Pleural fluid MORPHOLOGY Viable culture Biochemical identification Case report
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不同取材方法及检测技术在重症肺炎病原学诊断中的应用 被引量:1
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作者 刘龙群 唐敏 +2 位作者 张维维 吴可人 杨艳 《黑龙江医学》 2024年第3期271-273,共3页
目的:不同取材方法标本的合格率、不同微生物检测技术结果阳性率有差异,探究不同取材方法及检测技术在重症肺炎病原学诊断中的应用,以期提高病原学诊断的准确性。方法:回顾性分析2018年1月—2020年12月中国人民解放军联勤保障部队第九... 目的:不同取材方法标本的合格率、不同微生物检测技术结果阳性率有差异,探究不同取材方法及检测技术在重症肺炎病原学诊断中的应用,以期提高病原学诊断的准确性。方法:回顾性分析2018年1月—2020年12月中国人民解放军联勤保障部队第九〇四医院收治的123例重症肺炎患者的临床资料,同时收集患者经鼻或气管插管留痰培养、肺泡灌洗液培养、肺泡灌洗液宏基因组二代测序技术(mNGS)病原检测等数据,对标本的合格率及阳性率进行分析。结果:123例患者中经鼻或者气管插管留痰标本合格率70%、病原菌生长41例(33.3%),肺泡灌洗液标本合格率92%、病原菌生长76例(61.8%)、肺泡灌洗液mNGS病原菌核酸片段检出107例(86.9%)。经抗感染、抗休克、器官支持等治疗后,115例患者治愈出院,8例死亡。结论:肺泡灌洗液mNGS可以提高重症肺炎患者病原诊断阳性率,可为临床治疗提供依据。 展开更多
关键词 重症肺炎 肺泡灌洗液 宏基因组二代测序技术 痰培养
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mNGS在肺部感染病原学中的诊断价值及应用分析
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作者 王英盼 厉银平 《中国医学工程》 2024年第4期25-28,共4页
目的探讨宏基因组新一代测序(mNGS)在肺部感染病原体中的诊断价值及应用分析。方法选取2022年1月至2022年12月孝感市中心医院呼吸与危重症医学科收治的肺部感染(由常见的院内细菌如金黄色葡萄球菌、肺炎克雷伯杆菌、铜绿假胞单菌、大肠... 目的探讨宏基因组新一代测序(mNGS)在肺部感染病原体中的诊断价值及应用分析。方法选取2022年1月至2022年12月孝感市中心医院呼吸与危重症医学科收治的肺部感染(由常见的院内细菌如金黄色葡萄球菌、肺炎克雷伯杆菌、铜绿假胞单菌、大肠埃希菌、鲍曼不动杆菌等,病毒如单纯疱疹病毒、人类疱疹病毒7型等,真菌如白色念珠菌、曲霉菌等引起)患者258例作为研究对象。分别采集患者的肺泡灌洗液标本进行病原微生物mNGS检测与常规痰培养的检测,评估mNGS的诊断价值与应用分析。结果纳入研究的258例患者中,有200例患者通过mNGS检测得出,检出率为77.52%。有58例患者进行常规痰培养检测得出,检出率为22.48%。由此得出mNGS检测的病原微生物总检出率高于进行常规痰培养检测(P<0.05)。结论mNGS对肺部感染患者的病原微生物检出率高于常规痰培养检测,应用价值较高。但是具有一定的局限性。mNGS无法确定它所检测的序列是来自活菌还是死菌,同时也不能区分定植菌与致病菌。另外,mNGS的费用较高,临床上可酌情使用。 展开更多
关键词 宏基因组新一代测序 常规痰培养 肺部感染 病原菌 肺泡灌洗液
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肺泡灌洗液tNGS技术在下呼吸道感染患者中的应用
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作者 李培根 白英连 +2 位作者 黄秋惠 黄华 汪明星 《中国卫生标准管理》 2024年第17期126-130,共5页
目的探讨肺泡灌洗液多重靶向扩增-高通量测序法(targeted next-generation sequencing,tNGS)技术在下呼吸道感染患者中的应用效果。方法选取2022年1月—2023年5月于广东省连州市人民医院住院的176例下呼吸道感染患者为研究对象,所有患... 目的探讨肺泡灌洗液多重靶向扩增-高通量测序法(targeted next-generation sequencing,tNGS)技术在下呼吸道感染患者中的应用效果。方法选取2022年1月—2023年5月于广东省连州市人民医院住院的176例下呼吸道感染患者为研究对象,所有患者均接受肺泡灌洗液tNGS检测、灌洗液细菌培养、灌洗液结核分枝杆菌及利福平耐药检测(Xpert)、灌洗液半乳甘露聚糖检测(曲霉抗原,glactomannan,GM试验)、灌洗液真菌培养。比较5种检测方法对不同病原体的检出率,分析肺泡灌洗液tNGS检测下不同病原体分布特点,并对肺泡灌洗液tNGS检测和灌洗液细菌培养检测一致性分析。结果肺泡灌洗液tNGS检出率为92.05%,高于灌洗液细菌培养检测的22.16%,Xpert检测的9.09%,GM试验的16.48%以及灌洗液真菌培养的8.52%,差异有统计学意义(P<0.05)。球菌中检出率前3位为肺炎链球菌、粪肠球菌、金黄色葡萄球菌;杆菌中检出率前3位为流感嗜血杆菌、溶血嗜血杆菌、大肠埃希菌;真菌主要为白念珠菌和肺孢子菌;其他病原体主要为结核分枝杆菌;在常见呼吸道病毒中,疱疹病毒较为常见,其次为甲型流感病毒和鼻病毒;肺泡灌洗液tNGS检测和灌洗液细菌培养结果、灌洗液真菌培养存在一致性偏差。结论肺泡灌洗液tNGS检测技术可提高下呼吸道感染病原体检出率,该检测方式可作为下呼吸道感染病原体检测的有效检测方法,为临床治疗提供指导。 展开更多
关键词 下呼吸道感染 肺泡灌洗液 呼吸道病原体靶向测序 灌洗液细菌培养 灌洗液半乳甘露聚糖检测 灌洗液真菌培养
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不同条件下小鼠肺泡巨噬细胞体外培养的实验研究
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作者 云宏芳 蒋慧 +1 位作者 牛春晓 张纪岩 《中国免疫学杂志》 CAS CSCD 北大核心 2024年第6期1121-1125,共5页
目的:应用文献报道的肺泡巨噬细胞(AMs)体外培养方法,利用GM-CSF、TGF-β和PPAR-γ激动剂罗格列酮(简称GTR)培养体系,分析不同年龄小鼠、不同来源前体细胞产生AM样细胞或AMs的特征。方法:收取不同周龄小鼠支气管肺泡灌洗液(BALF)、骨髓... 目的:应用文献报道的肺泡巨噬细胞(AMs)体外培养方法,利用GM-CSF、TGF-β和PPAR-γ激动剂罗格列酮(简称GTR)培养体系,分析不同年龄小鼠、不同来源前体细胞产生AM样细胞或AMs的特征。方法:收取不同周龄小鼠支气管肺泡灌洗液(BALF)、骨髓单个核细胞以及胚胎15~17 d小鼠肝脏单个核细胞,种入12孔板,置于GTR体系中培养。流式细胞术检测产出细胞AM表面标志分子F4/80、CD11b、CD170和CD11c表达。结果:培养后,光镜下BALF来源细胞基本呈圆形,骨髓来源细胞近半呈圆形,胎肝来源细胞85%左右呈圆形。4周龄小鼠BALF-AMs比例接近90%,比例和数量均随小鼠年龄增加而增加,12周龄小鼠BALF-AMs数量接近1×106个。4周龄小鼠骨髓源AM样细胞比例不到20%,8周龄和12周龄小鼠比例为40%~45%,数量随小鼠年龄增加而增加,12周龄小鼠骨髓源AM样细胞数量约为0.8×106个/孔。胎肝源AM样细胞比例约为84%,1个孔产出的AM样细胞约为4×106个。结论:不同年龄小鼠、不同来源前体细胞产生AM样细胞或AMs的纯度和数量有较大差别,应根据具体研究要求选择合适方法。 展开更多
关键词 肺泡巨噬细胞 体外培养 肺泡灌洗液 骨髓 胎肝
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基于利福平耐药结核分枝杆菌半巢式实时荧光定量PCR技术检测胃液结核分枝杆菌在儿童肺结核中的诊断价值
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作者 阮树松 欧维正 祁仁杰 《当代医学》 2024年第14期134-138,共5页
目的分析基于利福平耐药结核分枝杆菌半巢式实时荧光定量PCR技术(Xpert-MTB/RIF)检测胃液结核分枝杆菌在儿童肺结核中的诊断价值。方法选取2020年1月到2021年12月贵阳市公共卫生救治中心儿童结核科收治50例疑似肺结核患儿作为研究对象,... 目的分析基于利福平耐药结核分枝杆菌半巢式实时荧光定量PCR技术(Xpert-MTB/RIF)检测胃液结核分枝杆菌在儿童肺结核中的诊断价值。方法选取2020年1月到2021年12月贵阳市公共卫生救治中心儿童结核科收治50例疑似肺结核患儿作为研究对象,取所有患儿晨起空腹胃液,分别行抗酸染色涂片、Xpert-MTB/RIF及罗氏(L-J)培养检测。以MTB培养作为金标准,分析不同检测方法结果,比较不同检测方法对儿童肺结核的诊断效能,分析Xpert-MTB/RIF及L-J培养阳性患儿MTB利福平耐药性情况及Xpert-MTB/RIF检测法与L-J培养法的药敏试验一致性。结果50例疑似肺结核患儿中,MTB培养结果检出36例肺结核,肺炎14例。抗酸染色涂片法检出阳性1例,阴性49例;L-J培养法检出阳性11例,阴性39例;Xpert MTB/RIF检测法检出阳性18例,阴性32例。Xpert MTB/RIF检测法、L-J培养法对儿童肺结核的诊断灵敏度高于抗酸染色涂片检测法,差异有统计学意义(P<0.05),Xpert MTB/RIF检测与L-J培养法诊断灵敏度比较差异无统计学意义。不同检测方法对儿童肺结核的诊断特异度、阳性预测值、阴性预测值比较差异均无统计学意义。Xpert MTB/RIF检测MTB对利福平的耐药敏感结果与L-J培养法的一致性Kappa值为0.891,具有较高的一致性。结论Xpert MTB/RIF检测较方便与快捷,具有较高的诊断效能,其检测MTB对利福平的药敏试验结果与L-J培养法一致性较高,有利于临床及早诊断儿童肺结核疾病并进行干预指导。 展开更多
关键词 肺结核 胃液 利福平 结核分枝杆菌 抗酸染色涂片 罗氏培养
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成年小鼠触液神经元体外分离培养及自我更新能力的鉴定
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作者 上官泽宇 陈婵娟 +5 位作者 李琦哲 谭伟 颜海健 王春庆 豆晓伟 李青 《中国组织工程研究》 CAS 北大核心 2025年第13期2728-2735,共8页
背景:课题组前期成功在体外分离培养乳鼠触液神经元,尚无研究报道有效分离培养高纯度成年小鼠触液神经元的方法,且触液神经元的自我更新能力是否随着年龄发生变化尚无研究。目的:建立一种高纯度成年小鼠触液神经元体外分离培养的方法,... 背景:课题组前期成功在体外分离培养乳鼠触液神经元,尚无研究报道有效分离培养高纯度成年小鼠触液神经元的方法,且触液神经元的自我更新能力是否随着年龄发生变化尚无研究。目的:建立一种高纯度成年小鼠触液神经元体外分离培养的方法,并鉴定成年小鼠触液神经元与乳鼠触液神经元在体外的自我更新能力。方法:从成年3月龄小鼠颈髓分离含有触液神经元的原代细胞贴壁培养并利用融合多模态成像基因的慢病毒转染细胞,通过嘌呤霉素筛选得到高纯度成年小鼠触液神经元细胞,在完全培养基中悬浮培养。通过免疫荧光检测成年小鼠触液神经元表达神经干细胞标记物巢蛋白(Nestin)及SOX2情况,观察成年小鼠触液神经元体外成球与传代能力;将同等数量(5×10^(3))的第3代成年小鼠及乳鼠触液神经元在同等条件下分为2组,分别接种在含有完全培养基的超低黏附培养板中,在体积分数5%CO_(2),37℃恒温箱悬浮培养,通过体外成球、CCK8实验、qPCR和Western blot鉴定成年小鼠及乳鼠触液神经元的自我更新能力。结果与结论:①实验成功在成年小鼠体内分离出高纯度触液神经元,在体外表达Nestin及SOX2,能形成神经球并连续传代。②成年小鼠触液神经元体外自我更新能力较乳鼠相比明显减弱,细胞培养到第4天时乳鼠触液神经元已经形成直径约为150μm的神经球,而成年小鼠触液神经元所形成的神经球直径仅为40μm(P<0.0001)。③CCK8增殖实验结果表明,成年小鼠触液神经元的增殖活性在培养后各时间点显著弱于乳鼠(P<0.0001)。④qPCR和Western blot检测发现成年小鼠触液神经元Nestin及SOX2的mRNA(P<0.0001)和蛋白表达量(P<0.01)较乳鼠显著下降。⑤上述结果证实,成年小鼠触液神经元的体外自我更新能力显著弱于乳鼠。 展开更多
关键词 脊髓损伤 成年小鼠触液神经元 内源性神经干细胞 自我更新能力 体外培养 细胞提纯 融合多模态成像基因的慢病毒 干细胞潜能鉴定
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基于Illumina平台mNGS技术在老年社区获得性肺炎肺泡灌洗液病原学诊断中的应用 被引量:1
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作者 李兴明 谢秀芳 +5 位作者 王宪刚 李云 罗进 何珍 邓魏岚 胡晓菁 《分子诊断与治疗杂志》 2023年第11期1965-1968,共4页
目的 探讨基于Illumina平台宏基因组二代测序(mNGS)技术在老年社区获得性肺炎(CAP)肺泡灌洗液病原学检查中的应用。方法 选取2022年1月至2022年6月内江第一人民医院收治的疑诊老年CAP患者200例为研究对象,比较肺泡灌洗液mNGS病原体检测... 目的 探讨基于Illumina平台宏基因组二代测序(mNGS)技术在老年社区获得性肺炎(CAP)肺泡灌洗液病原学检查中的应用。方法 选取2022年1月至2022年6月内江第一人民医院收治的疑诊老年CAP患者200例为研究对象,比较肺泡灌洗液mNGS病原体检测与传统细菌培养鉴定法对老年CAP的诊断价值。结果 200例患者经病原学检查共检出阳性154例(77.00%),其中细菌感染119例(59.50%)、病毒感染59例(29.50%)、真菌感染30例(15.00%)、肺炎支原体感染11例(5.50%)、肺炎衣原体感染7例(3.50%),包含混合感染50例(25.00%);mNGS共检出阳性183例(91.5%),其中细菌感染127例(63.50%)、病毒感染96例(48.00%)、真菌感染37例(18.50%)、肺炎支原体感染29例(14.50%)、肺炎衣原体感染15例(750%),包含混合感染75例(37.50%);mNGS阳性检出率、混合感染检测率较传统细菌培养高,差异有统计学意义(χ^(2)=15.845、7.723,P<0.05);mNGS检测时间较传统细菌培养鉴定短,差异有统计学意义(t=86.888,P<0.05);以临床诊断为金标准,mNGS诊断老年CAP的灵敏度、特异度为97.31%、85.71%,传统细菌培养灵敏度、特异度为71.43%、80.64%。结论 基于Illumina平台mNGS对病原体具有较高的检出率,检测时间更短,检测灵敏度、特异度更好,可以为老年CAP临床诊断提供重要依据。 展开更多
关键词 肺泡灌洗液 基因组二代测序 ILLUMINA 社区获得性肺炎 细菌培养鉴定
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宏基因组二代测序技术在神经外科颅内感染诊断中的应用价值 被引量:2
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作者 于海东 朱东 +1 位作者 高麟 李海龙 《临床医药实践》 2023年第11期803-804,833,共3页
目的:分析宏基因组二代测序技术(mNGS)在神经外科颅内感染病原学诊断中的应用价值。方法:选择2020年1月—2022年12月收治的20例神经外科术后拟诊颅内感染患者为研究对象,两周后由3位神经外科专家共同判断是否确诊为颅内感染。对20例患... 目的:分析宏基因组二代测序技术(mNGS)在神经外科颅内感染病原学诊断中的应用价值。方法:选择2020年1月—2022年12月收治的20例神经外科术后拟诊颅内感染患者为研究对象,两周后由3位神经外科专家共同判断是否确诊为颅内感染。对20例患者的脑脊液细菌培养结果和宏基因组二代测序检测结果进行对比。结果:宏基因组二代测序诊断神经外科术后颅内感染的灵敏度、特异度及准确率分别为100.0%(13/13),85.7%(6/7)及95.0%(19/20);细菌培养的灵敏度、特异度及准确率分别为69.2%(9/13),57.1%(4/7),65.0%(13/20);宏基因组二代测序的灵敏度、特异度及准确率均高于脑脊液细菌培养,差异有统计学意义(P<0.05)。结论:宏基因组二代测序技术在神经外科颅内感染病原学诊断中具有一定价值。 展开更多
关键词 宏基因组二代测序技术 神经外科颅内感染 脑脊液细菌培养
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614例孕中期羊水细胞染色体核型分析
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作者 杜静 唐登燕 +3 位作者 赵佳 王娇 张兵 周晓萍 《西部医学》 2023年第10期1550-1554,共5页
目的探讨孕中期羊水细胞染色体核型分析结果及不同产前诊断指征下异常核型检出率。方法纳入2018年12月—2022年3月在川北医学院附属南充市中心医院行羊膜腔穿刺术的614例孕妇染色体核型分析结果进行回顾性分析。对产前诊断指征进行分类... 目的探讨孕中期羊水细胞染色体核型分析结果及不同产前诊断指征下异常核型检出率。方法纳入2018年12月—2022年3月在川北医学院附属南充市中心医院行羊膜腔穿刺术的614例孕妇染色体核型分析结果进行回顾性分析。对产前诊断指征进行分类总结,并对不同产前指征下正常、异常核型进行比较,分析不同产前诊断下羊水细胞培养及染色体核型结果对临床遗传咨询价值。结果614例羊水样本中共检出33例异常核型,检出率为5.37%。异常核型中染色体数目异常(含嵌合体)21例,结构异常11例,同时出现数目及结构异常的1例。高龄、产筛高风险、B超异常、NT>2.5 mm、不良孕产史及用药史、无创DNA高风险、夫妻一方染色体异常这7类产前诊断指征中的异常核型检出率分别为2.53%、3.89%、2.7%、12.82%、3.03%、42.11%、40%。异常核型检出率在无创DNA高风险及夫妻一方染色体异常这两个指征中最高,二者间差异无统计学意义(P>0.05);NT>2.5 mm指征排第三;其余四种指征异常核型检出率最低,四种指征间差异无统计学(P>0.05)。结论羊水细胞培养及染色体核型分析技术对预防缺陷儿出生具有重要意义。产前诊断指征中,无创高风险与夫妻一方染色体异常指征对羊水异常核型的检出率最高。 展开更多
关键词 产前诊断 染色体核型分析 羊水细胞培养 异常核型检出率
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