鼻咽癌患者唾液中亚硝酸根含量的研究

目的研究鼻咽癌患者唾液中亚硝酸根含量。方法选择成人鼻咽癌患者26例为实验组,健康人27例为对照组,测定实验组和对照组所有人员唾液中亚硝酸根的含量。结果对照组唾液中亚硝酸根含量为(37.16±2.55)μmol/L,男女间无显著性差异。实验组唾液中亚硝酸根含量为(61.36±7.26)μmol/L,明显高于对照组。结论鼻咽癌患者唾液中亚硝酸根含量显著高于健康人。     

临床标本中非发酵菌的分布及耐药性分析

目的了解临床标本中非发酵菌的分布及耐药状况,为合理使用抗菌药物提供依据。方法各种标本经分离培养,用ATB Expression细菌鉴定仪鉴定,用ATB PSE药敏试条进行药敏试验,并作统计分析。结果7 395份标本分离出非发酵1 106株,分离率为14.9%,以铜绿假单胞菌的分离率最高(45.7%),各种临床标本中以痰液标本的检出率最高;除嗜麦芽寡养单胞菌和脑膜脓毒金黄杆菌外,对亚胺培南的耐药率最低,其次为头孢哌酮/舒巴坦。结论非发酵菌在各种临床标本中分布不同,种类较多,各种细菌之间耐药性差异较大,临床应及时采集标本,作病原学检测及药敏试验,并根据药敏试验结果选择抗菌药物,以减少细菌耐药产生,控制医院感染。     

Oxidative DNA damage and repair in children exposed to low levels of arsenic in utero and during early childhood: Application of salivary and urinary biomarkers

The present study aimed to assess arsenic exposure and its effect on oxidative DNA damage and repair in young children exposed in utero and continued to live in arsenic-contaminated areas. To address the need for biological specimens that can be acquired with minimal discomfort to children, we used non-invasive urinary and salivary-based assays for assessing arsenic exposure and early biological effects that have potentially serious health implications. Levels of arsenic in nails showed the greatest magnitude of difference between exposed and control groups, followed by arsenic concentrations in saliva and urine. Arsenic levels in saliva showed significant positive correlations with other biomarkers of arsenic exposure, including arsenic accumulation in nails (r = 0.56, P < 0.001) and arsenic concentration in urine (r = 0.50, P < 0.05). Exposed children had a significant reduction in arsenic methylation capacity indicated by decreased primary methylation index and secondary methylation index in both urine and saliva samples. Levels of salivary 8-OHdG in exposed children were significantly higher (~ 4-fold, P < 0.01), whereas levels of urinary 8-OHdG excretion and salivary hOGG1 expression were significantly lower in exposed children (~ 3-fold, P < 0.05), suggesting a defect in hOGG1 that resulted in ineffective cleavage of 8-OHdG. Multiple regression analysis results showed that levels of inorganic arsenic (iAs) in saliva and urine had a significant positive association with salivary 8-OHdG and a significant negative association with salivary hOGG1 expression.     

应用iTRAQ技术构建胃癌唾液的蛋白质组差异表达谱

目的：应用同位素标记相对和绝对定量（iTRAQ）技术标记定量蛋白组技术对胃癌患者唾液的全组蛋白进行鉴定和定量分析,初步获得胃癌患者唾液的蛋白组差异表达图谱。方法：胃癌患者与正常人唾液各10例,各组等量混合后利用iTRAQ8标试剂标记后的样本,采用Nano—LC—Ms／Ms分离、分析肽段,运用Proteinpilot4．0软件对蛋白进行鉴定和定量分析,并比较这些蛋白的表达差异。结果：对样品进行了2次Nano—LC—Ms／Ms,标记率〉95％,错误发现率〈1％,鉴定出符合假阳性率〈1％,共鉴定了747个蛋白。与正常对照组相比,胃癌组共出现了2倍以上表达差异的蛋白质21个,其中上调的12个,下调的9个。结论：iTRAQ联合NanoLC—MS／MS技术能高通量地筛选胃癌患者唾液中相关的蛋白,为胃癌患者早期诊断和预后提供可能的生物学标志物或治疗靶点,建立一种无创、简便、快捷实用的检测评估手段。     

Application of Saliva Inhibition to Detect Underlying Alloantibodies in Bombay Blood Group

IntroductionThe Bombay phenotype is a rare blood group determined by the absence of H antigens. Bombay individuals produce anti-H, a clinically significant antibody that react against all ABO blood group. Anti-H can mask underlying alloantibody during antibody investigation, a challenge in current transfusion practice. The aim of this article is to explore saliva inhibition, a novel method to detect underlying alloantibody in Bombay individuals.Case PresentationThe case is a 93-year-old female transfused with pre-donated autologous blood for a surgery. We determined anti-H subclass and thermal amplitude, secretor status, and optimal ratio of saliva and Bombay plasma. Plasma samples containing anti-H were spiked with anti-Fy(a) to determine the effectiveness of saliva inhibition in uncovering underlying alloantibodies.ResultsAnti-H was confirmed to be predominately IgM with broad thermal amplitude. Tube immediate spin (IS) showed stronger anti-H reactivity compared to column agglutination technology (CAT). Spiked anti-Fy(a) was successfully detected using saliva inhibition method.ConclusionTube IS appears more sensitive to anti-H. Saliva inhibition appears to be a promising method to detect underlying alloantibody in the plasma of Bombay phenotype individuals.     

Free light chains as an emerging biomarker in saliva: Biological variability and comparisons with salivary IgA and steroid hormones

BackgroundSalivary free light chains (FLCs) are an emerging biomarker in health and behavioural research. However, little is known regarding biological variability of salivary FLCs and how they relate to other established salivary biomarkers. This study aimed to investigate the diurnal and day-to-day variation of salivary FLCs and their relationship with salivary IgA and steroid hormones.MethodsA total of 46 healthy adults participated in studies exploring the biological variability of FLCs. Diurnal variation was investigated by collecting saliva samples immediately upon waking, 0.5 h, 3 h, 6 h, 9 h and 14 h post-waking. Saliva samples were assessed for FLCs, IgA, cortisol and dehydroepiandrosterone (DHEA). Between-day variation in FLCs and IgA was assessed by collecting saliva samples immediately upon waking for seven consecutive days. Participants underwent a dental examination to exclude oral health as a potential confounding variable. Within and between-person day-to day variation was explored in relation to a range of different factors: awakening time, sleep, exercise, well-being and alcohol consumption.ResultsSalivary secretion rates of FLCs decreased following waking and up to 3 h post-waking and then plateaued. This same pattern was observed for IgA. DHEA was stable upon waking and higher levels were seen in the morning with significantly lower levels thereafter. Cortisol levels significantly increased 0.5 h post-waking then continued to decline across the day. FLCs were significantly correlated with IgA but not cortisol or DHEA. Both FLCs and IgA parameters showed day-to-day variability, with coefficients of variation ≥ 40%. Earlier waking time was significantly correlated with higher FLC and IgA secretion rates. Inter-person differences in saliva parameter variability were observed but the degree of variation in FLCs and IgA was related within person. Inter-person day-to-day variation appeared to be uninfluenced by lifestyle or behavioural factors.ConclusionsSaliva FLCs secretion exhibits diurnal fluctuation that mirrors IgA fluctuation. Findings strongly indicate salivary FLC secretion is orchestrated by local plasma cells. FLCs and IgA both showed notable variability day-to-day, which was similar within person and influenced by awakening time. FLCs offer a promising adjunct to IgA in the measurement of oral immune activation.     

经脐腹腔镜乙状结肠肿瘤切除并脐再造术

目的探讨经脐腹腔镜乙状结肠肿瘤切除并脐再造术的可行性和临床应用前景。方法分析2011年1月～2012年12月成都市第二人民医院普外科同一治疗小组完成20例经脐腹腔镜乙状结肠肿瘤切除并脐再造术患者的手术时间、术中出血量、吻合口漏、切口感染、术后恢复情况及肿瘤根治情况等。结果 20例病人均在腹腔镜下顺利完成手术,手术时间（212.05±13.90）min,术中出血量（125.50±28.92）ml,淋巴结清扫数目（13.70±1.26）个,肠道功能恢复时间（3.10±0.64）d;术后切口脂肪液化1例,吻合口漏1例;术后平均8.7天出院。手术切口瘢痕隐蔽不易察觉,新＂肚脐＂形态良好。结论经脐腹腔镜乙状结肠肿瘤切除并脐再造术创伤小,恢复快,操作不复杂,安全性高,腹部美容效果明显,肿瘤根治性可靠。     

玻璃体和视网膜相关组织标本取材及应用研究现状

准确的玻璃体、视网膜相关组织标本取材及保存方法是保证临床诊断及开展严谨基础研究的基础。玻璃体标本取材的临床用途包括微生物培养、细胞学检测、变性性疾病检测、PCR分析、液基细胞学检测细胞形态等;实验研究用途包括DNA基因分析、蛋白定量分析、代谢物检查、RNA含量定量分析、细胞因子测定等。视网膜标本取材主要用于视网膜增生膜的PCR分析、免疫组织化学染色、免疫荧光检查、微血管密度评价以及细胞分离和培养等。了解玻璃体视网膜手术标本的取材和相关检测技术、材料的应用,可为玻璃体视网膜疾病的诊断提供更全面的思路及相关基础研究提供更广泛的参考。     

国内未见分布蝇种:澳洲麻蝇的形态和DNA条形码鉴定

目的 应用DNA条形码与形态分类相结合的方法,快速、准确鉴定在中山口岸入境船舶截获的1只蝇类.方法 2014年4月30日中山出入境检验检疫局神湾办事处从澳大利亚入境载有鲜葡萄的货船上截获1只蝇类,做成针插标本,并拍照存档.进行形态学鉴定,然后取1条后足提取基因组DNA,采用动物DNA条形码的通用引物(LCO1490和HCO2198)扩增目的片段,并进行纯化、克隆测序和序列分析,与NCBI及BOLD中的序列进行比对,构建NJ树.结果 形态学鉴定截获蝇类为雌性麻蝇科种类,由于缺少关于雌性麻蝇鉴定特征描述的参考资料,无法将该蝇鉴定到种,所获得的DNA条形码序列与BOLD中的澳洲麻蝇[Sarcophaga australis (Johnston et Tiegs,1921)]的序列相似度达100％.经过科技查新,确认是中国未见分布种.结论 根据鉴定结果,判定所截获的蝇类为雌性澳洲麻蝇,是我国首次截获.应用DNA条形码与形态鉴定相结合可以准确快速鉴定雌性麻蝇种类.     

Epigenetic age estimation in saliva and in buccal cells

Age estimation based on epigenetic markers is a DNA intelligence tool with the potential to provide relevant information for criminal investigations, as well as to improve the inference of age-dependent physical characteristics such as male pattern baldness or hair color. Age prediction models have been developed based on different tissues, including saliva and buccal cells, which show different methylation patterns as they are composed of different cell populations. On many occasions in a criminal investigation, the origin of a sample or the proportion of tissues is not known with certainty, for example the provenance of cigarette butts, so use of combined models can provide lower prediction errors.In the present study, two tissue-specific and seven age-correlated CpG sites were selected from publicly available data from the Illumina HumanMethylation 450 BeadChip and bibliographic searches, to help build a tissue-dependent, and an age-prediction model, respectively. For the development of both models, a total of 184 samples (N = 91 saliva and N = 93 buccal cells) ranging from 21 to 86 years old were used. Validation of the models was performed using either k-fold cross-validation and an additional set of 184 samples (N = 93 saliva and N = 91 buccal cells, 21–86 years old).The tissue prediction model was developed using two CpG sites (HUNK and RUNX1) based on logistic regression that produced a correct classification rate for saliva and buccal swab samples of 88.59 % for the training set, and 83.69 % for the testing set. Despite these high success rates, a combined age prediction model was developed covering both saliva and buccal cells, using seven CpG sites (cg10501210, LHFPL4, ELOVL2, PDE4C, HOXC4, OTUD7A and EDARADD) based on multivariate quantile regression giving a median absolute error (MAE): ± 3.54 years and a correct classification rate ( %CP±PI) of 76.08 % for the training set, and an MAE of ± 3.66 years and a %CP±PI of 71.19 % for the testing set. The addition of tissue-of origin as a co-variate to the model was assessed, but no improvement was detected in age predictions. Finally, considering the limitations usually faced by forensic DNA analyses, the robustness of the model and the minimum recommended amount of input DNA for bisulfite conversion were evaluated, considering up to 10 ng of genomic DNA for reproducible results. The final multivariate quantile regression age predictor based on the models we developed has been placed in the open-access Snipper forensic classification website.