论著

急性肺损伤相关分子标志物的鉴定及临床意义探索

Identification and clinical significance of molecular biomarkers associated with acute lung injury

:245-254
 
目的 通过公共数据库筛选急性肺损伤(ALI)及急性呼吸窘迫综合征(ARDS)相关分子标志物,并探索其临床意义。方法 利用基因表达综合数据库(GEO)中有关ALI/ARDS基因表达芯片研究的两个数据集GSE76293和GSE10474,通过STRING网站和Cytoscape软件对差异基因进行蛋白互作网络分析并筛选ALI/ARDS相关关键基因。采用A549细胞构建ALI模型,并通过转录组测序验证关键基因在细胞中的表达差异情况。结果 2个GEO数据集中共筛选出共同上调基因27个,共同下调基因26个。主要参与抗原加工和外源抗原递呈、免疫受体活性调节、内质网膜构成等生物学功能,且与抗原加工、细胞分化等信号通路有关。蛋白互作网络分析共筛选出10个ALI/ARDS相关关键基因,分别为CD4、HLA-DQB1、CD74、HLA-DRA、FCGR2B、TOR1A、RELA、NME8、RNF19B、RHOB。细胞转录组测序结果显示,关键基因的上调或下调特征及表达差异情况与GEO数据集分析结果一致。结论 CD4等关键基因可能参与ALI/ARDS发生、发展的生物学过程,是ALI/ARDS临床诊断及预后预测的潜在个体化分子标志物。
Objective To identify molecular biomarkers associated with acute lung injury(ALI)/ acute respiratory distress syndrome(ARDS)and to explore their clinical significance with public databases. Methods Two datasets GSE76293 and GSE10474 in Gene Expression Omnibus(GEO)database for ALI/ARDS gene expression chip study were used to screen genes with significant differences in both datasets.The protein-protein interaction(PPI)analysis of co-expression genes was performed based on the STRING website and Cytoscape software,and then key genes related to ALI/ARDS were identified with cytoHubba method.The ALI model was constructed using A549 cells cultured in vitro,and the expression differences of key genes in the cells were verified by RNA sequencing. Results A total of 27 up-regulated genes and 26 down-regulated genes were screened in both the two GEO datasets with Venn Diagramm.These co-expression genes were mainly involved in biological functions such as antigen processing and presentation of exogenous peptide antigen,immune receptor activity,integral component of lumenal side of endoplasmic reticulum membrane and were related to signal pathways such as antigen processing and cell differentiation.A total of 10 key genes(CD4,HLA-DQB1,CD74,HLA-DRA,FCGR2B,TOR1A,RELA,NME8,RNF19B,RHOB)related to ALI/ARDS were identified. The results of cell RNA sequencing showed that the up-regulated or down-regulated characteristics and expression differences of key genes were consistent with the results of GEO datasets. Conclusions Several key genes identified in this study may be involved in the biological process of ALI/ARDS development,and may be potential individualized molecular markers for clinical diagnosis and prognosis prediction of ALI/ARDS.
论著

抗-M抗体影响血型鉴定及对临床输血的影响

:606-610
 
目的 探究抗-M抗体影响血型鉴定及对临床输血的影响,分析不规则抗体筛查必要性,以期进一步提升临床输血的安全性及有效性。方法 分析2020年1月—2023年4月于蚌埠医学院第二附属医院输血科检查的100例O型红细胞对照管凝集患者样本资料,使用全自动血型仪微柱凝胶卡、反定型试管法对样本进行血型鉴定和抗体检查,使用抗人球蛋白实验检测抗体类型及特异性,并分析抗-M抗体在不同环境温度下的效性。结果 血型仪微柱凝胶法显示100例标本中存在正反定型不符89.00%,正反定型相符11.00%;反定型试管法复核显示100例标本A、B、O细胞管均为阳性,对照管均为阴性。对89例正反定型不符的样本予以分析,抗-M是最主要影响抗体,占比56.18%;IgM、IgG是最主要影响抗体类型;13例未鉴定出抗体特异性,占比14.61%。89例正反定型不符的样本中男25例、女64例;71例存在妊娠史或输血史,占比79.78%,其中的11例合并妊娠史和输血史。50例抗-M抗体标本在不同气温下呈现不同效价,在4 ℃、22 ℃下会出现凝集梯度;在37 ℃下,50例样本中反应强烈31例,占比62.00%,阴性19例,占比38.00%。结论 抗-M类不规则抗体是影响血型鉴定、威胁输血安全的主要因素,临床需加强输血前血型鉴定,明确抗-M抗体临床检测的意义,通过多种途径提升鉴定准确性,对合并高危险因素的患者更需要着重关注,保障输血安全。
论著

兔腰椎间盘严重退变骨水泥成形术模型建立与鉴定

Establishment and identification of cementoplasty model of rabbit lumbar disc with severe degeneration

:35-39
 
目的 建立兔腰椎间盘严重退变骨水泥成形术模型并进行鉴定。方法 选用新西兰白兔6只,手术干预前摄腰椎正侧位X线片并进行MRI扫描Pfirrmann分级,之后通过腹外斜肌与腰大肌间隙入路手术去除兔腰2~3椎间盘髓核组织及部分纤维环模拟腰椎间盘严重退变状态。饲养6周后相应腰椎节段椎间盘进行MR扫描Pfirrmann分级,确认相应腰椎节段椎间盘符合严重退变影像表现后再次手术显露相应椎间隙并注入骨水泥。1周后再次摄腰椎正侧位X线片并行MRI扫描Pfirrmann分级,终末处死并解剖动物检查椎间盘内骨水泥填充情况。结果 兔腰椎间盘退化模型建立6周后磁共振Pfirrmann分级为Ⅴ级。椎间隙骨水泥注射后1周其术后磁共振Pfirrmann分级为Ⅳ。骨水泥注射模型1周后拍摄手术节段X线片显示骨水泥较好地填充于腰2~3间隙,椎间隙高度接近正常状态。终末处死并解剖动物发现腰椎节段椎间盘内骨水泥填充良好无脱落或松动。结论 通过腹外斜肌与腰大肌间隙入路,手术去除椎间盘髓核组织及部分纤维环6周后,往椎间隙内注入骨水泥,可获得较为可靠的新西兰大白兔腰椎间盘严重退变骨水泥成形术的动物模型。
Objective To establish and identify the rabbit model of lumbar disc with severe degeneration.Methods Six New Zealand white rabbits were selected,lumbar X-ray and Pfirrmann grade by MR scan were performed before surgical intervention.Along the space of obliquus externus abdominis and psoas major,the front edge of L2 to L3 was exposed.Then,the nucleus pulposus and part of annulus fibrosus were removed to imitate severe degeneration of lumbar disc.After 6 weeks of rearing,the operated lumbar disc was graded by MR scan,confirming that the lumbar disc met the image of severe degeneration,and then exposed the intervertebral space and injected bone cement.One week later,the anterior lumbar X-ray and the MRI scan for Pfirrmann grading were taken.The animals were sacrificed and dissected to check the bone cement filling in the intervertebral disc.Results The rabbit MR Pfirrmann grade of intervertebral disk was V after 6 weeks of first operation.One week after intervertebral cement injection,the MR Pfirrmann grade was Ⅳ.The surgical segment X-ray was taken one week after the cement injection,which showed that the cement was well filled in the L2-L3 gap and the vertebral space height was close to normal.Animals were sacrificed and dissected,the lumbar intervertebral disc was well filled with cement without shedding or loosening.Conclusions A reliable animal model of lumbar disc with severe degeneration in New Zealand white rabbits can be obtained by injecting cement into the intervertebral space after 6 weeks of removal of the intervertebral disc nucleus pulposus and part of the annulus fibrosus through the obliquus externus abdomins and psoas major intervertebral space.
论著

VITEK-MS系统直接鉴定体液培养阳性标本的研究

Direct bacterial identification in positive culture bottles by use of the VITEK-MS system

:11-13
 
目的 利用基质辅助激光解吸电离飞行时间质谱系统(VITEK-MS)对体液培养阳性瓶进行直接鉴定,探索快速诊断临床致病菌的新策略。方法 收集体液培养阳性瓶,不经琼脂平板培养,直接利用VITEK-MS进行鉴定,并与传统生化鉴定的方法进行比较分析。结果 50例体液培养阳性瓶中,传统细菌鉴定法检出47株阳性菌,3例阴性;而VITEK-MS直接鉴定法检出31株阳性菌,同样3例阴性。VITEK-MS直接鉴定法灵敏度达65.96%,特异度为100%,临床符合率为68%。鉴定时间从24小时缩短到2小时。结论 利用VITEK-MS质谱系统直接鉴定体液培养阳性标本中的病原菌,能有效缩短细菌鉴定时间,准确快速地诊断临床致病菌。
Objective To find a fast method for detection of pathogens in positive culture bottles by using the VITEK-MS system. Methods VITEK-MS microbial identification system was used to directly identify the bacteria in the positive culture bottles, without culture on agar plates. The identification results were further compared with those by the traditional biochemical identification. Results Forty-seven bacterial strains were identified by traditional biochemical methods among 50 positive culture bottles, and 3 of them were negative. Of these 50 samples, thirty-one bacterial strains were identified by VITEK-MS and 3 were also negative. The sensitivity and specificity for direct VITEK-MS identification were 65.96% and 100%, and the clinical coincidence rate was 68%. The turn around time for identification was reduced from 24 to 2 hours. Conclusion Direct identification of bacterial pathogens in positive culture bottles by VITEK-MS could reduce turn around time, and lead to accurate and fast diagnosis.
论著

RD105缺失基因检测法用于青海省北京/W系结核分枝杆菌鉴定

Identification of Beijing/W lineage mycobacterium tuberculosis in Qinghai province by using an RD105 deletion test

:11-14
 
目的 了解青海省北京/W系结核分枝杆菌分布特征。方法 收集青海地区结核分支杆菌临床分离株,采用RD105缺失基因检测鉴定北京/W系结核分枝杆菌。结果 共收集237株结核分枝杆菌临床分离株,采用RD105缺失基因检测鉴定北京/W系结核分枝杆菌220株,占92.8%,非北京/W结核分枝杆菌,共17株,占7.2%。北京/W系结核分枝杆菌在青海地区性别与民族分布差异没有统计学意义(P>0.05)。结论 北京/W结核分枝杆菌为青海地区流行菌株,在人群易于发生感染和传播。
Objective To ascertain the epidemiological characteristics of Beijing/W lineage Mycobacterium tuberculosis in Qinghai Province. Methods M. tuberculosis clinical isolates were collected and identified with an RD105 deletion test.Statistical analysis was performed by using the test. Results Totally, 237 clinical isolates of M. tuberculosis were collected in which 220 strains (92.8%) belonged to the Beijing/W lineage of M. tuberculosis while 17strains (7.2%) belonged to the non-Beijing/W lineage of M. tuberculosis according to the RD105 deletion test. There were no significant differences in the distribution of Beijing/W lineage of M. tuberculosis in the gender and nationality (P>0.05). Conclusion Beijing/W lineage of M. tuberculosis were prevalent in Qinghai province and prone to having infection and transmission in the crowd.
论著

输血前不规则抗体鉴定结果分析与临床输血安全

Pre-transfusion irregular antibody identification results and clinical blood transfusion safety

:1087-1093
 
        目的   明确输血前各抗体的分布特点,探讨自身抗体和同种抗体在性别、年龄、输血史、妊娠史和不同疾病中的差异,并根据抗体血型血清学特性制定个体输血方案,以确保临床输血安全。方法    选取2021年6月—2024年8月在广东省第二人民医院输血科申请输注红细胞或手术备血的29 662例患者,采用低离子抗人球蛋白微柱凝胶法进行不规则抗体筛查,结果阳性的标本经科内讨论并送广州血液中心血型参比实验室进行抗体鉴定,通过统计血液中心回报结果分析各抗体的特异性。结果  29 662例患者标本中不规则抗体结果为阳性的有208例,阳性率为0.70%。同种抗体占比47.69%,Rh、MNS和Lewis红细胞血型系统共占同种抗体中的94.50%,其中常见意外抗体:抗-E占31.87%、抗-M占14.29%、抗-Mur占19.78%、抗-C占7.69%和抗-e占7.69%。同种抗体与自身抗体在性别、年龄、妊娠史等方面比较差异无统计学意义(P0.05);在输血史及不同科室疾病等方面比较差异有统计学意义(P<0.05)。结论    输血前进行输血相容性检测是必要的,应对拟申请红细胞的患者进行不规则抗体筛查,阳性者宜进行抗体鉴定,明确其抗体的特异性及临床意义,以确保临床输血安全。
       Objective  To clarify the distribution characteristics of each antibody before transfusion,to explore the differences between autoantibodies and homologous antibodies in gender,age,history of blood transfusion,history of pregnancy and different diseases,and to formulate individual transfusion protocols based on the serological characteristics of antibody blood groups to ensure the safety of clinical blood transfusion.Methods  A total of 29 662 patients who applied for red blood cell transfusion or surgical blood preparation in the hospital from June 2021 to August 2024 were selected for irregular antibody screening by low-ion anti-human globulin microcolumn gel method.The samples with positive results were discussed within the department and sent to the Blood Type Reference Laboratory of Guangzhou Blood Center for antibody identification.The specificity of each antibody was analyzed by blood center reported results.Results  Among 29 662 patients,208 were positive for irregular antibody,the positive  rate was 0.70%.The alloantibodies accounted for 47.69%,Rh,MNS and Lewis erythrocyte blood group system accounted for 94.50% of alloantibodies,among which the common unexpected antibodies were anti-E 31.87%,anti-M 14.29%,anti-MUR 19.78%,anti-C 7.69% and anti-E 7.69%.There were no significant differences between alloantibodies and autoantibodies in gender,age and pregnancy history(P>0.05).There were statistically significant differences in blood transfusion history and diseases in different departments(P<0.05).Conclusions  It is necessary to test the compatibility of blood transfusion before transfusion.Screening for irregular antibodies should be carried out in prospective transfusion patients,and antibody identification should be carried out in positive patients to clarify the specificity and clinical significance of their antibodies to ensure the safety of clinical transfusion.
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