临床常见分离菌对不同配比头孢哌酮/舒巴坦的药敏差异

目的了解临床常见病原菌对不同配比头孢哌酮/舒巴坦药敏纸片的耐药性,为临床合理选择头孢哌酮/舒巴坦提供依据。方法收集某院2014年上半年临床分离病原菌1 141株,采用纸片扩散法检测不同病原菌对头孢哌酮/舒巴坦两种药敏纸片(70/35 μg与75/75 μg)的抗菌活性。结果1 141株病原菌中肠杆菌科细菌675株（59.16%）,非发酵菌447株(39.18%),其他革兰阴性杆菌19株(1.66%)。对头孢哌酮/舒巴坦两种药敏纸片(70/35 μg、75/75 μg)的耐药率：超广谱β 内酰胺酶（ESBLs）阳性大肠埃希菌(eco,221株)分别为7.69%、2.26%,肺炎克雷伯菌(kpn,92株)分别为10.87%、3.26%;亚胺培南耐药鲍曼不动杆菌（IRAB 295株）分别为54.92％、11.19％;两种药敏纸片抗菌活性比较,差异均有统计学意义（P＜0.05）。ESBLs阴性肠杆菌科细菌（eco,135株;kpn,98株）、亚胺培南敏感鲍曼不动杆菌（51株）、铜绿假单胞菌（48株）、嗜麦芽窄食单胞菌（22株）对两种头孢哌酮/舒巴坦药敏纸片的耐药率比较,差异均无统计学意义（均P>0.05）。结论ESBLs阳性肠杆菌科细菌与IRAB对不同配比头孢哌酮/舒巴坦药敏纸片的抗菌活性具有差异,临床治疗此类细菌引起的感染时要充分考虑头孢哌酮/舒巴坦的剂型,以达到预期治疗效果。     

福尔马林固定石蜡包埋样本全基因组文库构建流程优化

目的优化建立适合本实验室针对人源福尔马林固定石蜡包埋(Formalin-Fixed and Paraffin-Embedded, FFPE)样本的全基因组(Genomic DNA, gDNA)文库构建方案。 方法探讨并设计了在FFPE全基因组文库构建各环节优化条件，分析比较FFPE DNA文库片段大小及文库转化率，以选择最优的文库构建流程。 结果最终确定DNA起始量50 ng、20 min片段化酶切时间，采用4×稀释的接头进行20 min连接，经过2次特定比例的磁珠筛选方案进行文库构建，实现片段主峰在300 bp~400 bp左右，文库转化率达60%。 结论本操作流程针对FFPE样本可以保证高质量的文库构建，同时实现较高文库转化效率。     

新生儿罕见胎儿肺间质肿瘤一例

报道1例罕见的新生儿肺间质肿瘤。男性新生儿,出生1 d因呼吸困难就诊,影像学诊断新生儿肺炎,左胸腔占位,疑先天性肺隔离症和肺囊肿。病变行手术切除,术后病理检查和免疫组织化学诊断为胎儿肺间质肿瘤(FLIT)。该报道旨在引起儿科和病理医师对新生儿FLIT的认识、关注和研究,以积累更多的循证医学数据,避免不适当诊断。     

miR-200c inhibits metastasis of breast cancer cells by targeting HMGB1

miR-200c has been shown to regulate the epithelial-mesenchymal transition （EMT） by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software （miRanda） was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB 1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of＇miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.     

高危型HPV载量与宫颈细胞学病变相关性分析

目的通过高危型人乳头状瘤病毒(HR-HPV)载量与不同宫颈细胞学病变相关性分析,进一步研究病毒载量在HR-HPV阳性女性中的分流作用。方法选取2016年1月至2018年9月济南金域医学检验中心接检的6337例宫颈细胞学和HR-HPV双检测进行宫颈癌筛查女性作为研究对象。回顾性分析4431细胞学判读为无上皮内病变或恶性病变(NILM),1284例细胞学判读为低级别鳞状上皮内病变(LSIL),622例判读为高级别鳞状上皮内病变(HSIL)和鳞状细胞癌(SCC)(≥HSIL)女性与高危型HPV病毒载量间的相互关系。结果HR-HPV阳性率在NILM、LSIL和≥HSIL中分别为6.3%、83.5%和92.4%,呈明显增高趋势,差异具有统计学意义(P<0.001)。HR-HPV阳性的NILM和细胞学检查异常(LSIL和≥HSIL组)女性在低、中和高病毒载量组中的分布趋势差异具有统计学意义(P<0.001)。低、中和高载量组中,HR-HPV阳性的NILM女性呈明显降序排列,差异具有统计学意义(P<0.001),而LSIL和≥HSIL女性呈明显升序排列,差异具有统计学意义(P<0.001),但LSIL和≥HSIL女性分布差异无统计学意义(P=0.625)。同时,HR-HPV阳性LSIL和≥HSIL组的平均病毒载量,在<30岁组、30岁~49岁组、≥50岁组和总人群中,均要明显高于NILM组(均P<0.05),但LSIL和≥HSIL组间均无明显差异(均P>0.05)。结论不同病毒载量对区分细胞学筛查正常者和异常者有较好价值,但对LSIL和≥HSIL病变区分无意义。     

Coinheritance of OLFM2 and SIX6 variants in a Chinese family with juvenile-onset primary open-angle glaucoma: A case report

BACKGROUNDJuvenile-onset primary open-angle glaucoma (JOAG), characterized by severe elevation of intraocular pressure and optic neuropathy prior to the age of 40, is a rare subtype of primary open-angle glaucoma. Several genetic mutations have been associated with JOAG.CASE SUMMARYThe proband patient was a young male, diagnosed with primary open-angle glaucoma at the age of 27. The patient and his unaffected parents who have been excluded from classic genetic mutations for primary open-angle glaucoma were included to explore for other possible genetic variants through whole genome sequencing and bioinformatics analysis. In this trio, we found two heterozygous variants inherited from the parents in the proband: c.281G>A, p.Arg94His in OLFM2 and c.177C>G, p.Ile59Met in SIX6. Both genetic mutations are predicted through bioinformatics analysis to replace evolutionary conserved amino acids, therefore rendering a pathogenic effect on proteins. In contrast, very low frequencies for these genetic mutations were recorded in most common control databases.CONCLUSIONThis is the first report on coinherited mutations of OLFM2 and SIX6 in a JOAG family, which shows the complexity of JOAG inheritance. Large-scale clinical screening and molecular functional investigations on these coinherited mutations are imperative to improve our understanding of the development of JOAG.     

遗传病二代测序临床检测全流程规范化共识探讨(1)——遗传检测前流程

遗传检测前的准备工作是基因检测的基础和出发点,其流程包括临床信息收集、检测方案拟定、检测前遗传咨询以及知情同意书和检测委托书的填写等。临床若能有效识别出遗传性疾病,可极大提高二代测序的诊断率,从而降低医疗成本,提高临床诊疗的功效。二代测序结果的分析在很大程度上依赖于对基因型-表型相关性的了解,全面准确地采集和评估临床表型,并用统一的标准术语来描述和记录尤为重要。不同类型的遗传病或突变种类需要用特定的检测手段方能事半功倍。检测前遗传咨询能够帮助患者及其亲属了解相关基因检测的意义并协助制定个体化的检测策略,为后续的跟踪随访奠定基础。     

山东东明县23 088名已婚女性宫颈癌筛查结果分析

目的分析山东省东明县已婚女性宫颈癌筛查结果,为该地区宫颈癌预防提供科学依据。 方法回顾性汇总分析2016年9月至2017年6月期间,于济南金域医学检验中心病理室行宫颈癌筛查的东明县23 088名已婚女性的细胞学检测结果。年龄范围为19~90岁,分为5组：<30岁组810例、30~39岁组5174例、40~49岁组9301例、50~59岁组5707例、≥60岁组2096例。同时对其中细胞学异常者随访,得到高危型人乳头状瘤病毒(high-risk human papillomavirus,hrHPV)检测结果61例和组织活检结果438例,进行分析。 结果23 088名已婚女性经细胞学检测,检出异常细胞学者1177例(5.1%),其中：不能明确判读的非典型鳞状细胞(atypical squamous cells of unknown significance,ASC-US)者729例(3.2%),不能排除高级别鳞状上皮内病变的非典型鳞状细胞(atypical squamous cells cannot exclude HSIL,ASC-H)/非典型腺细胞(atypical glandular cells,AGC)者43例(0.2%),低级别鳞状上皮内病变(low grade squamous intraepithelial lesions,LSIL)者289例(1.3%),高级别鳞状上皮内病变(high grade squamous intraepithelial lesions,HSIL)者116例(0.5%)。随访细胞学异常者hrHPV检测阳性率为50.0%(31/61例),组织活检阳性率为45.6%(200/438例)。不同年龄组间不同异常细胞学病变阳性率比较,ASC-US (χ²=5.502,P=0.019)和LSIL(χ²=3.956,P=0.047)阳性率存在明显差异。ASC-US以50~59岁组197例(3.5%)最高,30~39岁组134例(2.6%)最低;LSIL以<30岁组12例(1.5%)和30~39岁组76例(1.5%)最高,50~59岁组50例(0.9%)最低。不同细胞学病变的组织活检结果有明显差异。活检正常者(χ²=113.536,P<0.001)在ASC-US中所占比(75.4%)最高,而HSIL所占比(9.8%)最低;活检结果为宫颈上皮内瘤变1(cervical intraepithelial neoplasia,CIN1) (χ²=37.817,P<0.001)在LSIL所占比(44.4%)最高,而HSIL所占比(6.6%)最低;活检结果为CIN2/3/原位腺癌(adenocarcinoma in situ,AIS) /鳞状细胞癌(squamous cell carcinoma,SCC) (χ²=603.863,P<0.001)则在ASC-US中所占比(4.0%)最低,而HSIL所占比(83.6%)最高。不同细胞学病变(χ²=7.946,P=0.019)和组织学活检结果(χ²=3.925,P=0.048)的hrHPV阳性率也有明显差异。ASC-US/ASC-H/AGC、LSIL和HSIL的阳性率分别为34.5%、58.3%和87.5%;组织学活检正常、CIN1和CIN2/3/AIS/SCC的阳性率分别为37.5%、60.0%和75.0%。 结论细胞学筛查能有效地发现癌前病变和宫颈癌,且细胞学病变的严重程度、hrHPV的阳性率高低与组织学活检结果的严重程度有较好的一致性,这能为制定该地区宫颈癌筛查策略提供一定依据。     

基于分子检测技术的实验室质量改进与管理

临床医生获得与疾病相关的分子检测数据以及据此做出准确诊断,是后续实行精准治疗的前提和基础。分子检测结果的准确与否不仅要依靠先进检测设备和检测技术,还要高度重视实验室管理和数据的质量控制。高质量的分子检测数据是包括技术人员的综合专业素质、实验操作流程的规范性、实验室内部质量管理等方面集中反映。正确处置这些要素对检测结果的准确性和可重复性起着重要作用。本文就核酸分子检测中常用的突变扩增阻滞系统(amplification refractory mutation system,ARMS)技术结合实时荧光定量PCR(ARMS-PCR)检测技术和二代测序技术在应用过程中出现的问题进行分析和讨论,旨在与业内人士一起学习交流,以提高检测结果的可靠性,为临床提供可靠的实验室诊疗依据。