目的 基于液相芯片技术建立一种可同时快速检测6种临床常见血流感染厌氧菌的方法.方法 通过GenBank寻找脆弱拟杆菌、厌氧消化链球菌、具核梭杆菌、中间普雷沃菌、产气荚膜梭菌和痤疮丙酸杆菌的16S rDNA基因序列,经序列比对分析,选择菌内保守菌间特异的区域作靶序列,使用Primer 5.0软件设计特异性引物及探针,建立不对称多重PCR扩增体系,将PCR产物与偶联核酸探针的荧光微球混合物进行杂交,建立一种多重检测方法,并对该方法的特异度、灵敏度和重复性进行评价.收集2020年6月～2022年5月东莞市厚街医院阳性血厌氧培养瓶84例及阴性培养瓶16例,应用建立的方法进行检测,结果与传统培养法进行比较.结果 6种厌氧菌靶基因序列经PCR扩增后,电泳成像清晰可见6条目标条带;经反应体系优化,生物素标记与非标记引物浓度比为4:1时,选择退火温度56℃进行多重扩增,加入5μl PCR产物,52℃温育20 min检测效果最佳;各目标序列荧光强度中位值(median fluorescence intensity,MFI)>1000,无非特异性信号;最低检测限可达103cfu/ml～104cfu/ml;当菌液浓度为105cfu/ml时,批内及批间的变异系数分别在7.38％和11.10％以下;100例临床样本中6种厌氧菌的检测结果与培养法一致.结论 成功建立一种快速、简便、高效的液相芯片方法,可同时检测6种常见血流感染厌氧菌.

目的 对乙型肝炎病毒脱氧核糖核酸(HBV DNA)荧光定量检测方法进行性能验证及评价.方法 参考《医学实验室质量和能力认可准则在分子诊断领域的应用说明(CNAS-CL36)》及美国临床和实验室标准化协会(CLSI)EP系列文件的相关要求对实验室HBV DNA荧光定量PCR检测方法的精密度、测量正确度、抗干扰能力、线性范围和检测下限等进行验证及评价.结果 HBV DNA定量检测低值(1×103 U/mL)、高值(1×106 U/mL)标本的批内.CV分别为0.07％和0.06％,批间CV分别为0.11％和0.08％;正确度误差在允许范围内;干扰物(三酰甘油、胆红素、血红蛋白)样本结果与对照样本的偏差≤±lg0.4;在1.45×102～1.45×109 U/mL范围内线性关系良好(Y=1.042X-0.337,R2=0.997);检测下限可以达到100 U/mL.结论 HBV DNA荧光定量检测方法的各项性能指标与厂家声明相符,可用于临床常规检测.

目的 建立一种可同时、快速检测2型糖尿病患者降糖药相关药物基因多态性的液相芯片检测系统.方法 根据7个目的基因的rs号,在Genbank中查找其靶位点附近碱基序列(目的基因包括磺脲类受体1,转录因子7类似物2,胰岛素受体底物1,过氧化物酶体增殖物激活受体γ,有机阳离子转运蛋白与多药和有毒化合物排出家族,有机阴离子转运蛋白家族成员1B1等),以PrimerPlex软件设计等位基因特异性引物,Primer6.0软件设计含检测位点的PCR引物,通过多重PCR扩增,等位基因特异性引物延伸(ASPE),MagPlex～Tag微球杂交,液相芯片系统Luminex 200检测荧光信号,确定基因型,优化反应体系并以5份代表性标本进行方法学评价.收集2019年1月～12月东莞市厚街医院新诊2型糖尿病患者血液样本115例,采用建立的系统检测上述7个靶位点,并随机选取25例与测序结果比较.结果 7个目标基因经PCR扩增,产物电泳成像后清楚可见7条目标条带,无非特异性条带;经优化ASPE杂交条件,选择退火温度55℃,Biotion～dCTP浓度与dCTP浓度的比值为3:1,37℃孵育45 min时检测效果最佳;临床样本检测结果显示:纯合子荧光强度中位值(median fluorescence intensity,MFI)比率均>0.9或<0.1,杂合子MFI比率在0.4～0.6之间.纯合子MFI比率批内批间CV在0.9％～3.3％和2.1％～4.6％,而杂合子则在2.9％～7.3％和5.2％～11.2％;DNA最低检测限为0.75ng;25例样本的检测结果与测序结果完全一致,准确度100％.结论 该研究成功建立了一种新的液相芯片检测系统,高效便捷,能同时检测7个目的基因型,满足临床的需求.

Objective To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P4502C9 (CYP2C9), CYP2C19, CYP4F2, Vitamin K epoxide reductase (VKORC1) and ATP-binding cassette subfamily B member1 (ABCB1) gene, which were associated with warfarin and clopidogrel, based on liquid phase chip technology. Methods Method establishment. The eight gene sequences near targeted sites related to warfarin and clopidogrel were found in Genbank, and the specific primers and probes were designed. Through multiple PCR amplification, followed by allele specific primer extension (ASPE), and MagPlex-Tag microspheres hybridization, the suspension array Luminex 200 system step-by-step, the genotypes were determined by fluorescence signal. The reaction system was optimized and its methodological evaluation was performed. 260 patients with antithrombotic therapy from Dongguan houjie hospital were recruited in this study form June 2017 to December 2018. The eight genotypes of the 260 patients were detected by the established method, and the results were compared with the sequencing results. Results The results of 260 samples showed that allelic median fluorescence intensity (MFI) ratios of homozygotes (mutant/wild-type) were all greater than 0.9 or less than 0.1, and all the allelic MFI ratios of heterozygotes were between 0.3 and 0.6. The within run and between run coefficients of variance for allelic MFI ratios were lower than 6.4％and 10.9％, respectively. The minimum DNA template requirements was 0.75ng. The genotypes of 260 patients determined by the established method were completely concordant with the sequencing results. Conclusion A method was established successfully for rapid detecting the genotypes which associated with warfarin and clopidogrel based on liquid phase chip technology.

目的 了解本地区妇女宫颈脱落细胞人乳头瘤病毒(HPV)的感染情况和基因型分布特点,为临床防治措施的制定提供参考依据.方法 回顾性分析2015年3月至2016年6月于本院行宫颈脱落细胞HPV检测阳性患者资料,统计HPV的感染率、基因型分布特点、年龄变化趋势等.结果 本次统计从131 1名患者中检测出26种HPV基因型1773例,其中高危型占67.84％,低危型占32.16％,前五位高危型HPV基因型分别是HPV-52,16,58,53和39,分别占12.97％,10.38％,6.15％,5.92％和5.02％,前五位低危型HPV基因型分别是HPV-6,43,81,61和11,分别占7.90％,4.85％,3.95％,3.89％和3.38％.结论 本地区HPV感染以单一高危型为主,最常见的基因亚型为HPV52、16、6、58和53,且发病有年轻化趋势,值得关注.

We compared urinary liver-type fatty acid-binding protein (L-FABP) among non-pregnant and pregnant women with and without gestational diabetes mellitus (GDM). Higher urinary L-FABP was found in pregnant with and without GDM, and considerably higher urinary L-FABP was found in the GDM group compared with the non-GDM group. Hyperglycemia and anemia were related with high urinary L-FABP expression.

目的 模拟糖尿病条件,观察人肾小管上皮细胞(HK-2)生长情况和Smad1表达的变化,探讨高糖对肾小管上皮细胞的损伤机制.方法 用DMEM/F12细胞液培养HK-2细胞,分为对照组,中糖组(12.5mmol/L葡萄糖),高糖组(25mmol/L葡萄糖),分别培养24h、48h、72h后倒置显微镜下观察HK-2的形态和数量变化,采用蛋白印迹技术检测Smad1的蛋白表达.结果 高糖组刺激72h后,与对照组相比HK-2细胞数为(105.333±6.429)×105/mL,而对照组HK-2细胞数为(50.333±2.51)×105/mL,差异有统计学意义(P＜0.01);高糖组HK-2细胞形态改变,Smadl蛋白表达明显增加.结论 高糖能够诱导肾小管上皮细胞数量及形态发生改变,并且这一作用可能与Samd1表达增加有关.

Objective To investigate the strains distribution and resistance characteristics of pathogenic bacteria isolated from blood culture and provide basis for clinical medication. Methods Blood samples were cultivated and continuously monitored by VersaTREK automated blood culture system with the matching blood culture bottles,and the positive bottles were transferred timely. The identification of bacteria and susceptibility test were performed by MicroScan A/S-4 bacterial automatic analyzer. Results A total of 237 strains of pathogenic bacteria were isolated from 2460 blood culture samples. The top five strains included CNS, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Enterococcus. Positive pathogens were mainly distributed in ICU, general surgical department and respiratory medicine department. Susceptibility tests showed that Gram negative bacilli were highly sensitive to imipenem and Gram positive cocci had no resistance to vancomycin and linezolid. Conclusion It is important to comprehend the blood culture results promptly for the selection of antimicrobial agents and has great significance to improve cure rate.

Objective To explore the associations between different HBeAg staus ,serum HBV DNA level and hepatic injury in patients with chronic hepatitis B (CHB) .Methods HBV DNA level and serum ALT were detected by quantitative Real‐time PCR and velocity methods separately .The degree of hepatic injury was determined by the level of ALT ,and the associations between HBV DNA level and hepatic injury both in HBeAg negative CHB and HBeAg positive CHB patients were analyzed accordingly .Re‐sults Elder age and lower HBV DNA level were found in HBeAg negative CHB patients than in HBeAg positive patients (P<0 .01) ,however ,no difference of hepatic injury were found between the two groups (P>0 .05) .Serum HBV DNA level was posi‐tively correlated with the degree of hepatic injury in HBeAg negative CHB patients (P<0 .05) but no similar correlation was found in HBeAg positive patients .Conclusion Positive serum HBeAg is an effective indicator of HBV replication and no correlation is presented between HBV DNA level and intrahepatic inflammation in HBeAg positive CHB patients .However ,HBV DNA level is closely correlated with liver damages in HBeAg negative CHB patients ,more attention should be paid to clinical treatment and fol‐low‐up monitoring of HBeAg negative CHB patients .

Objective To explore the relationship between urinary smad1 and diabetes nephropathy in T2DM patients.Methods The 118 T2DM patients and 28 control subjects were enrolled in the trial.The patients were divided into 3 groups according to urine albumin/creatinine ratio(UACR),64 in normal albuminuria group,36 in microalbuminuria group,and 18 in macroalbuminuria group.Their urinary smad1 was measured by ELISA.FPG,creatinine(Cr),serum cystatin C(CysC),HbA1c,and C-P were also examined.Glomerular filtration rate(GFR) was determined by Macisaac formulae to analyze the relations between urinary smad1 and the common clinical indices.Results The level of urinary smad1 was significantly higher in the T2DM patients than in the controls and positively related with UACR in T2DM(R=0.412,P=0.000),and was getting progressively higher in the normal albuminuria group,microalbuminuria group,and macroalbuminuria group.Pearson analysis showed that the level of urinary smad1 was positively related with FPG and HbA1c,but negatively with fasting C-P.Logistic regrsssion analysis showed that the urinary smad1 was an independent risk factor for diabetic nephropathy in T2DM patients.Conclusion Urinary smad1 is related with the degree of diabetic nephropathy,and may be a promising marker for the diagnosis and monitoring the treatment of DN.

Objective To explore the early diagnostic significance of serum and urine neutrophil gelatinase associated lipocalin(NGAL)and urine kidney injury molecule-1(KIM-1)in ICU patients with acute kidney injury(AKI).Methods A total of 23AKI patients and 24non-AKI patients,hospitalized in ICU of this hospital from Jun.2010to May.2011,and 21healthy subjects were enrolled and detected for serum and urine NGAL and KIM-1,and serum urea nitrogen(BUN),creatinine(Cr)and cystatin C(Cys C).All data were statistically analyzed.Results Serum levels of NGAL,BUN,Cr and Cys C,and urinary levels of NAGL and KIM in AKI group were higher than healthy group(P<0.01).The differences of serum levels of NGAL,BUN and Cr,and urinary levels of NGAL and KIM-1between detection before and after therapy were significant(P<0.01).The differences of serum levels of NGAL and Cr,and urinary levels of NGAL and KIM-1between AKI group and non-AKI group were significant(P <0.01).Area under receiver operating characteristic curve of serum NGAL,urinary NGAL,urinary KIM-1,serum Cr,Cys C and BUN was 1.00,0.97,1.00,0.98,0.98and 0.99respectively.Conclusion The level of serum NGAL,urine NGAL and KIM-1could be increased significantly in AKI patients,which might be with early diagnostic value in AKI.

User-generated images are now prevalent across social media platforms, such as Facebook, Twitter, and various blogospheres. These images can be categorized and ranked based on their relevant topics. In this paper, we present and compare candidate schemes for mining salient images related to a specific topic or object among a large number of images from a blogosphere. Identifying salient images consists of several steps: calculating the similarity between images, k-means clustering images, and ranking images. In each step, we propose a set of alternatives and as a result, present an optimal combination scheme by conducting an empirical comparison of the performance of each scheme. Furthermore, to address scalability, we also present a distributed version of the schemes and experimental results based on MapReduce on top of a Hadoop environment.

Objective To explore the relationship between tubular markers and glomerular hyperfiltration(GHF)in type 2 diabetes mellitus(T2DM)patients.Methods Estimated glomerular filtration rate(eGFR)was calculated by Macisaac' s formulae based on cystatin C(CysC)levels.30 cases of T2DM with GHF,58 cases of T2DM with norm-GFR and 24 healthy people were recruited in the study.Creatinine,CysC,glucose and hemoglobin A1C were measured.Tubular markers NGAL,KIM-1 and NAG were determined by ELISA or colorimetric analysis.The correlation between these markers and eGFR were conducted by multivariate analysis in the GHF group.Results The GHF group had higher urinary NGAL,KIM-1 levels and lower serum NGAL level than those in other two groups(P ＜ 0.05).There was no statistical difference in the urinary NAG level among the three groups.Correlation analysis showed that eGFR was positively related to fasting blood glucose,HbA1c,urinary NGAL,and KIM-1(r=0.250,0.237,0.388,0.470),negatively related to serum NGAL(r=-0.159)in the GHF group.Conclusion The high levels of urinary tubular damage markers are found in T2DM patients with GHF,which may be a proof that GHF can directly result in early renal tubular damage.

The purpose of this study was to investigate the prevalence of tubular damage in short-term (less than five years) type 2 diabetes mellitus (T2DM) patients and to explore the correlation between tubular markers and their relationship with renal indices at different stages of diabetic nephropathy. A group of 101 short-term T2DM patients and 28 control subjects were recruited. Tubular markers, such as neutrophil gelatinase-associated lipocalin (NGAL), N-acetyl-β-d-glucosaminidase (NAG), and kidney injury molecule 1 (KIM-1), as well as urinary albumin excretion were measured in voided urine. Glomerular filtration rate (GFR) was estimated via Macisaac’s formula. The patients were further categorized into three groups, namely, the normoalbuminuria, microalbuminuria, and macroalbuminuria groups, according to their urine albumin/creatinine ratio (UACR). Urinary tubular markers were compared and their correlations with renal indices [UACR and estimated GFR (eGFR)] were analyzed among the different diabetic groups. Compared with the control group, Urinary NGAL [median (IQR)][83.6(41.4−138.7) μg/gcr vs. 32.9(26.1−64.5) μg/gcr], NAG [13.5(8.7−17.9) U/gcr vs. 7.6(6.5−13.0) U/gcr] and KIM-1 [120.0(98.4−139.9) ng/gcr vs. 103.1(86.8−106.2) ng/gcr] in the T2DM were all markedly increased. For all patients, urinary NGAL had stronger positive correlations with UACR than NAG (R = 0.556 vs. 0.305, both P < 0.05). In addition, only urinary NGAL showed a negative correlation with eGFR (R = −0.215, P < 0.05). Urinary KIM-1, however, showed no significant difference among the three T2DM groups and did not correlate with either UACR or eGFR. As UACR increased from the normoalbuminuria to the last macroalbuminuria group, all of the markers increased. However, only the concentrations of NGAL were statistically different among the three diabetic groups. The correlation between the tubular markers and their relationships with the renal indices differed markedly among the three T2DM groups. In conclusion, these results suggest that tubular damage is common in short-term T2DM patients. Urinary NGAL may be a promising early marker for monitoring renal impairment in short-term T2DM patients.

The aim of this study was to assess the relationship between urinary Smad1 and glomerular hyperfiltration (GHF) in type 2 diabetes mellitus (T2DM), and to explore the factors related to the urinary Smad1 in T2DM. The reference value of the estimated glomerular filtration rate (eGFR) was determined in 248 healthy individuals. 30 patients with GHF, 58 patients with norm-GFR T2DM, and 24 healthy patients who served as controls were recruited. Urinary Smad1, fasting plasma glucose (FPG), fasting serum C-Peptide (C-P), hemoglobin A1C (HbA1c), cystatin C, and other chemistry laboratory parameters of T2DM participants and controls were measured. Patients with GHF had higher levels of urinary Smad1 than the control group, and those with norm-GFR. For T2DM patients with body mass index, age, and gender adjustments, urinary Smad1 was positively correlated with FPG, HbA1C, and eGFR, but negatively correlated with fasting serum C-P. Multivariate linear regression analysis demonstrated that eGFR, HbA1C, and fasting serum C-P were independently associated with urinary Smad1. High levels of urinary Smad1 were found in GHF patients with T2DM, which may be another potential mechanism of GHF in relation to diabetic nephropathy.

Aim To assess whether glomerular hyperfiltration (GHF) could result in renal tubular damage in type 2 diabetes mellitus (T2DM) patients. Methods Reference value of estimated glomerular filtration rate (eGFR) was determined in 248 healthy individuals based on serum CysC levels. GHF was defined as an eGFR exceeding the sex-specific 97.5th percentile in non-diabetic individuals. In the present study, 30 with GHF, 58 with norm-GFR T2DM, and 24 healthy controls were recruited. Tubular markers, such as urinary N-acetyl-β-d-glucosaminidase (NAG) and kidney injury molecule 1 (KIM-1), as well as serum and urinary neutrophil gelatinase-associated lipocalin (NGAL), were measured and compared. The correlation of these markers with eGFR was analyzed in the GHF group. Results The GHF group had higher urinary NGAL and KIM-1 levels but lower serum NGAL level than the norm-GFR and control groups. Slightly decreased serum NGAL and increased urinary NGAL levels were also noted in the norm-GFR group compared with those of the controls. There was no statistical difference in the urinary NAG values among the three groups. Correlation analysis showed that eGFR was positively related to fasting blood glucose (FBG), HbA1c, urinary NGAL, and KIM-1, but negatively with serum NGAL in the GHF group. Conclusion Higher urinary tubular damage markers were found in T2DM patients with GHF than the norm-GFR and control groups, probably a direct proof that GHF is a deleterious factor for diabetic nephropathy.

Objective To analyse the relationships between microalbumin-to-creatinine ratio(UACR) and qualitative protein results in different urinary specific gravity,and investigate the influence of urinary specific gravity on the detection of urine protein by dry chemistry technology. Methods Qualitative protein analysis and urinary specific gravity(SG) in 844 urinary samples were conducted by dry chemistry technology,and both the urinary protein and creatinine were also measured by automatic biochemical analyzer in these samples. The samples were grouped into five groups according to their SG. The relationships between microalbumin-to-creatinine ratio(UACR) and qualitative protein results were analysed in different groups. Results For the samples with negative in protein detection, much higher false negative ratios were found in the lower SG group. The false negative ratio reduced from 34.1% to 2.7% when the urinary SG was ranged from ≤1.005 to ≥1.030 in the negative protein urinary samples..For the samples with positive in protein detection, on the other hand, the higher SG the samples, the more false positive were found. There was no false positive results found in the samples with SG ≤1.005. Conclusion Urinary qualitative protein analysis could be influenced by SG. So it was advised to test the UACR or 24 h-urinary protein analysis for the abnormal SG samples.

Objective To explore the prognostic value of Serum Cystatin C and APACHEⅡ scoring systems in Intensive Care Unit patients with acute kidney injury.Methods 32 patients with AKI from June 2010 to May 2011 were collected and analyzed,which conform to the criteria for the diagnosis of AKI.Recorded its cystatin C change,at the same time collecting clinical data,including gender,age,chronic disease history,24 h inside all sorts of assay index(routine blood,blood gas analysis,renal function,blood electrolyte,etc.) the worst value,and carried on the APACHE Ⅱ score.AKI group were divided into live group and death groups,observing the two groups of APACHE Ⅱ score difference.In addition the same period selected 20 cases of patients in ICU which did not happen AKI as a non AKI group.Results Of the 32 cases of acute kidney injury were analyzed,11 cases of death,mortality was 34.4%.With the APACHE II scores increased,Serum Cystatin C levels are rising.Survival and mortality between groups is statistically significant(P<0.05),logistic regression analysis shows that the higher of the serum Cystatin C and the APACHEⅡ ≥15,oliguria tied to mortality in patients with acute kidney injury(P<0.05).The above indicated the Cystatin C concentration plus APACHEⅡ ≥15 are the main factors related to prognosis of patients with acute kidney injury.Conclusion Serum Cystatin C and APACHE II scores are correlated,In addition of comprehensive therapy on Patients of ICU AKI,under the precondition of cystatin C rise combined with APACHE Ⅱ score ≥ 15 points can be used as evaluation index of the prognosis of patients with AKI.

近年来,我国2型糖尿病的发病率不断上升,由此引发的各种慢性微血管并发症也在增多,糖尿病肾病是糖尿病最常见的微血管并发症,也是终末期肾功能衰竭的主要原因.糖尿病肾病的确切发病机制不明,蛋白尿是最常见的临床表现,最近有研究认为肾小管功能紊乱在糖尿病肾病蛋白尿的形成中发挥着重要作用[1-2],NGAL作为肾小管损伤的重要指标,被认为是最有应用前景的肾损伤标志物之一[3-4].本研究旨在检测2型糖尿病患者血和尿NGAL含量,以探讨其在糖尿病肾病中的临床价值.