道地药材天麻DNA真伪鉴定试剂盒的研制与评价

目的 建立一种将DNA提取技术与PCR技术相结合的天麻DNA真伪鉴定试剂盒,并对试剂盒性能进行方法学评价。方法 通过美国国家生物技术信息中心(NCBI)查找天麻及其常见伪品紫茉莉根、大丽菊块茎、马铃薯的ITS2序列,应用DNAMAN进行多序列比对,NCBI-Primer-Blast设计天麻特异性引物。改良植物常用DNA提取的CTAB法,确保提取出高效的正品天麻及其常见伪品的基因组DNA,应用紫外分光光度法测其浓度及纯度。优化PCR反应体系,确定试剂盒的组成与反应条件,随机抽样检测市售天麻样品。结果 应用所研制的试剂盒提取样品DNA,纯度OD260 /OD280值为(1.87±0.13),灵敏度为10 ng·μL^-1,3次重复性检测结果相同,反复冻融 5、10、15、20次对检测效果无影响,于-20 ℃可保存1年,检测10个市售天麻样品,其中7个是正品,3个是伪品。结论 道地药材天麻DNA真伪鉴定试剂盒特异性强,灵敏度高,重复性和稳定性均良好,检测结果准确,适用于天麻及其常见伪品的快速鉴别。     

A retrospective evaluation of a multiplex polymerase chain reaction test directly applied to blood for the management of sepsis in the critically ill

Background Blood culture (BC) is the established gold standard for microbiological diagnosis of bloodstream infection (BSI); however, its sensitivity is poor. Objectives The primary objective was to determine the sensitivity and specificity of the Magicplex Sepsis Real-time Test, a multiplex polymerase chain reaction test (mPCR), and BC to detect BSIs. Secondary outcomes included determining the prevalence of BSIs. Methods A retrospective review of a technical evaluation of the mPCR. Patients requiring BC had a blood sample collected for mPCR Results The respective sensitivity and specificity of mPCR for the detection of BSI were 50% (n=7/14) and 58% (n=18/31), while the sensitivity and specificity using BC were 36% (n=5/14) and 68% (n=21/31), respectively. The addition of mPCR to BC increased BSI detection during sepsis from 36% to 64%. Conclusion The use of mPCR directly applied to blood may increase the detection of micro-organisms associated with BSIs in critically ill patients requiring BC investigation. Contributions of the study Our data add to a growing body of evidence indicating that mPCR applied directly to blood prior to incubation increases the detection of pathogenic bacteria among hospitalised patients for whom blood cultures are performed for suspected infection. Our study was performed in a low-to-middle income country with a higher sepsis prevalence, a greater burden of multidrug-resistant organisms and clinically defined sepsis. This strengthens the robustness and generalisability of this body of evidence.     

化疗对结肠癌Th1和Th2类细胞因子基因表达的影响及意义

目的探讨结肠癌患者体内Th1和Th2类细胞因子的基因表达水平及化疗对其表达的影响。方法收集67例结肠癌患者、35例正常人和30例结肠息肉患者血清。采用RT-PCR技术,检测3组及结肠癌组化疗后外周血单个核细胞(PBMC)Th1和Th2类细胞因子mRNA表达水平。结果正常对照及结肠息肉组Th1和Th2类细胞因子基因表达呈阴性或基本不表达。结肠癌组IL-4、IL-6、IL-10mRNA表达水平显著高于正常对照及结肠息肉组(P<0.05),且与结肠癌的分化程度及临床分期相关(P<0.05);IFN-γ无阳性表达(0/67),IL-2则仅有3例阳性表达,其表达与结肠癌的分化程度及临床分期无相关性(P>0.05)。化疗后Th1类细胞因子表达增强,Th2类细胞因子的表达则减弱(P<0.05)。结论结肠癌患者Th1和Th2类细胞因子表达失衡,细胞免疫受抑,化疗可使结肠癌患者Th2类细胞因子的强势表达向Th1类逆转。     

B类清道夫受体CD36的研究进展

CD36是一种跨膜糖蛋白,可与多种配基相结合,它在体内的分布广泛。作为一种清道夫受体,CD36既参与脂质的代谢,也在带负电荷的生物大分子黏附中发挥作用。对CD36的调控可能是巨噬细胞泡沫化和动脉粥样硬化形成的关键因素。迄今还发现其与肿瘤、肥胖等多种问题相关,一些CD36相关的药物研究也在进行中。     

GeXP多重RT-PCR诊断小儿肺炎支原体感染性脑炎的价值分析

目的 探讨GeXP多重RT-聚合酶链反应(polymerase chain reaction,PCR)诊断小儿肺炎支原体(mycoplasma pneumoniae,MP)感染性脑炎的价值.方法 采用回顾性研究,选取2017-05—2020-05郑州儿童医院收治的85例初诊为感染性脑炎患儿为研究对象,采集患儿脑脊液标本...     

Ossicular chain reconstruction in chronic otitis media: hearing results and analysis of prognostic factors

IntroductionThe goal of ossiculoplasty is to improve hearing and the success of this procedure depends on several factors.ObjectiveAnalyze the hearing results in patients with chronic otitis media undergoing ossicular chain reconstruction, as well as predictive factors for successful surgery.MethodsCharts of patients undergoing ossiculoplasty between 2006 and 2016 were reviewed. Sixty-eight patients were included, totaling 72 ears. The following data was analyzed: gender, age, smoking status, laterality, pathology, audiometric exams, type of surgery, previous surgery, characteristics of the middle ear, otorrhea and ossicular chain status. Patients were also classified according to two indices: middle ear risk index and ossiculoplasty outcome parameter staging. The results were evaluated by comparing the air-bone gap before and after surgery. The success of reconstruction was defined as air-bone gap ≤20 dB and the improvement of speech reception Thresholds, calculated through the mean frequencies 0.5, 1, 2 and 3 kHz.ResultsReconstruction success rate was 61%. The mean preoperative air bone gap was 34.63 dB and decreased to 17.26 dB after surgery. There was a correlation between low risk in middle ear risk index and ossiculoplasty outcome parameter staging indices with postoperative success. The most frequently eroded ossicle was the incus and the type of prosthesis most used was tragal cartilage. In the patients without incus, we achieved success in 74.2% of the surgeries. In the absence of the stapes, the success rate decreased to 63.3%. In the absence of the malleus, 85% of the patients had and air bone gap ≤20 dB.ConclusionWe achieved good audiometric outcomes in ossiculoplasty and the results are comparable to other centers. Ossicle status influenced postoperative results, especially in the presence of stapes. We also concluded that the indexes analyzed may help to predict the success of the surgery.     

Is there any association between Helicobacter pylori and otitis media with effusion?

IntroductionIt is proposed that Helicobacter pylori can be responsible for the development of otitis media with effusion.ObjectiveThe aim of this study is to investigate the prevalence of H. pylori in the adenoid tissue and fluid of the middle ear in patients who suffer from adenoid hyperplasia and otitis media with effusion in comparison with those who suffer from adenoid hyperplasia without otitis media with effusion.MethodsThis is a case–control study that was carried out in 50 children of age 2–7 years old who were admitted with adenoid hyperplasia. Patients were divided into case and control groups. The study group included 25 patients with adenoid hyperplasia and otitis media with effusion and the control group included 25 patients with adenoid hyperplasia without otitis media with effusion. The patients in both groups underwent surgical adenoidectomy. For the case group we carried out myringotomy and placement of tympanostomy tube, and fluid samples were collected under sterile conditions. The samples were sent to the laboratory for polymerase chain reactions.ResultsIn the case group H. pylori was found to be positive in 18 samples of the middle ear fluid (70%) and in 1 polymerase chain reaction adenoid tissue sample (4%). In the control group H. pylori was positive in 3 samples of adenoid tissues (12%). There was no gender difference.ConclusionH. pylori is one of the important bacteria that plays a role in the pathogenesis of otitis media with effusion. Whether adenoid tissue may be a reservoir for H. Pylori is unclear.     

The relationship between ANT1 and NFL with autophagy and mitophagy markers in patients with multiple sclerosis

Multiple sclerosis (MS) is the most common inflammatory neurodegenerative disease. Neurofilament light chain (NFL) is a novel adverting biomarker of axonal damage that suggested as a useful assistant in the monitoring of MS patients. It has been shown that the auto/mitophagy associated with MS pathogenesis. In this study, we aimed to study correlation between ATG5 and Parkin, as markers of autophagy and mitophagy respectively, with NFL and ANT1 in serum and cerebrospinal fluid (CSF) in MS subjects. ATG5, Parkin, NFL, and ANT1 levels were measured in a cross-sectional study of 40 MS patients compared with gender, age and BMI matching healthy volunteers. Based on our results, levels of ATG5, Parkin, and NFL significantly were elevated in both serum and CSF of MS patients comparing control individuals (p < 0.0001) but ANT1 levels significantly was decreased in both serum and CSF of MS patients comparing control individuals (p < 0.0001). The correlation indices between NFL, ANTI1, ATG5 and Parkin in both case and control groups showed a direct and moderate the correlation between ANTI1 and ATG5 in the CSF level of the control group (r = 0.554, P = 0.011). Our data support the feasibility of quantifying of NFL as a sensitive and clinically meaningful serum/CSF biomarker to follow-up nerve tissue injury in MS condition.