减毒猪霍乱沙门菌ΔcrpΔcyaΔasdC78-1(pYA3493)宿主-载体平衡致死系统的构建及其生物学特性研究

目的:开发应用于猪的稳定携带异源基因的口服活疫苗载体。方法:本研究在减毒猪霍乱沙门菌ΔcrpΔcya C78-1的基础上,运用自杀性质粒pREasd介导的细菌同源重组技术,构建缺失株ΔcrpΔcyaΔasd C78-1,再将携带asd基因的互补质粒p YA3493电转至上述菌株,构建ΔcrpΔcyaΔasd C78-1(p YA3493)宿主-载体平衡致死系统,并进一步研究其生物学特性。结果:PCR及测序结果共同表明ΔcrpΔcyaΔasd C78-1(p YA3493)构建成功。生物学特性检测结果表明ΔcrpΔcyaΔasd C78-1(p YA3493)的血清型与亲本株ΔcrpΔcya C78-1和疫苗株C500相同,并能够稳定遗传缺失型asd基因片段;其生长速度略慢于ΔcrpΔcya C78-1,且二者均明显慢于疫苗株C500;其生化特性结果也与ΔcrpΔcya C78-1基本相同。小鼠口服攻毒试验表明ΔcrpΔcyaΔasd C78-1(p YA3493)的毒力与ΔcrpΔcya C78-1基本相当,但其半数致死量约为疫苗株C500的412倍。结论:上述结果表明,减毒猪霍乱沙门菌ΔcrpΔcyaΔasd C78-1(p YA3493)有潜力作为高效表达外源基因的口服活疫苗载体。     

Prenatal Exposure to Phthalate Esters and Behavioral Syndromes in Children at 8 Years of Age: Taiwan Maternal and Infant Cohort Study

Background: Few studies have shown an association between prenatal phthalate exposure and adverse effects on neurodevelopment and behavior in young children.Objectives: We aimed to assess the relationship between prenatal exposure to phthalate esters and behavior syndromes in children at 8 years of age.Methods: A total of 122 mother–child pairs from the general population in central Taiwan were studied from 2000 to 2009. Mono-methyl phthalate (MMP), mono-ethyl phthalate (MEP), mono-butyl phthalate (MBP), mono-benzyl phthalate (MBzP), and three di-(2-ethylhexyl) phthalate (DEHP) metabolites—mono-2-ethylhexyl, mono-2-ethyl-5-hydroxyhexyl, and mono-2-ethyl-5-oxohexyl phthalates (MEHP, MEHHP, and MEOHP)—were measured in maternal urine collected during the third trimester of pregnancy using liquid chromatography–electrospray ionization–tandem mass spectrometry. Behavioral syndromes of children at 8 years of age were evaluated using the Child Behavior Checklist (CBCL). Associations between log₁₀-transformed creatinine-corrected phthalate concentrations and standardized scores of the CBCL were estimated using linear regression models or multinomial logistic regressions with adjustments for potential confounders.Results: Externalizing problem scores were significantly higher in association with a 1-unit increase in log₁₀-transformed creatinine-corrected concentrations of maternal MBP (β = 4.29; 95% CI: 0.59, 7.99), MEOHP (β = 3.74; 95% CI: 1.33, 6.15), and MEHP (β = 4.28 ; 95% CI: 0.03, 8.26) after adjusting for the child’s sex, intelligence, and family income. Meanwhile, MBP and MEOHP were significantly associated with Delinquent Behavior and Aggressive Behavior scores. The same pattern was found for borderline and/or clinical ranges.Conclusions: Our findings suggest positive associations between maternal DEHP and dibutyl phthalate (DBP) exposure and externalizing domain behavior problems in 8-year-old children.Citation: Lien YJ, Ku HY, Su PH, Chen SJ, Chen HY, Liao PC, Chen WJ, Wang SL. 2015. Prenatal exposure to phthalate esters and behavioral syndromes in children at 8 years of age: Taiwan Maternal and Infant Cohort Study. Environ Health Perspect 123:95–100; http://dx.doi.org/10.1289/ehp.1307154     

Glucose-regulated protein 78 regulates multiple malignant phenotypes in head and neck cancer and may serve as a molecular target of therapeutic intervention

Glucose-regulated protein 78 (Grp78) is an endoplasmic reticulum chaperone protein and is overexpressed in various cancers. However, it is unclear how significance of this molecule play an active role contributing to the oncogenic effect of head and neck cancer (HNC). To investigate the potential function of Grp78, six HNC cell lines were used. We found that Grp78 is highly expressed in all six cell lines and many of the proteins were localized in the periphery regions, implying other function of this molecule aside from endoplasmic reticulum stress response. Knockdown of Grp78 by small interfering RNA significantly reduced cell growth and colony formation to 53% to 12% compared with that of controls in all six HNC cell lines. Using in vitro wound healing and Matrigel invasion assays, we found that cell migration and invasive ability were also inhibited to 23% to 2% in all these cell lines tested. In vivo xenograft studies showed that administration of Grp78-small interfering RNA plasmid into HNC xenografts significantly inhibited both tumor growth in situ (>60% inhibition at day 34) and liver metastasis (>90% inhibition at day 20). Our study showed that Grp78 actively regulates multiple malignant phenotypes, including cell growth, migration, and invasion. Because knockdown Grp78 expression succeeds in the reduction of tumor growth and metastatic potential, this molecule may serve as a molecular target of therapeutic intervention for HNC. [Mol Cancer Ther 2008;7(9):2788-97].     

Increase diagnostic efficacy by combined use of fingerprint markers in mass spectrometry--plasma peptidomes from nasopharyngeal cancer patients for example

OBJECTIVES: There is no plasma marker for detecting nasopharyngeal cancer (NPC). We developed a bead-based affinity fractionated proteomic method to search potential plasma markers for NPC. DESIGN AND METHODS: Affinity purification of heparinized plasma with Cu-chelated beads and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis were used to screen potential NPC markers. We compiled MS protein profiles for 47 patients with NPC and compared them to profiles from 28 healthy controls. The spectra were statistically analyzed using flexAnalysis and ClinProt bioinformatics software. Diagnostic efficacy was evaluated by determination of the assay sensitivity and specificity of each marker. RESULTS: Twelve mass fingerprint markers differing between cancer and control spectra were found. The sensitivities of these NPC markers are various ranging from 36% to 83%, and the specificities were all over 90%. Combine use of these markers significantly increases diagnostic efficacy. In which, the combined markers (2020 Da and 4635 Da) possess best discriminator with high sensitivity (94%) and specificity (93%). We further identify a C3 fragment, C3f, that may serve as a biomarker for NPC. CONCLUSION: The combined use of mass fingerprint markers in plasma proteome will enhance diagnostic efficacy for NPC. This method can be applied to search for novel plasma markers for cancers.     

云南省宾川县片形吸虫病人群流行病学调查及危险因素分析

目的 目的 了解云南省宾川县人群片形吸虫感染情况及相关危险因素。 方法 方法 2014年2-3月, 选择宾川县州城镇 2个行政村的6个自然村作为调查点, 对 ≥ 5岁的常住人口采用ELISA法进行血清抗体检测, 采用Kato?Katz法和尼龙绢集卵孵化法对血检阳性者进行粪便虫卵检查。对筛检出的片形吸虫病病例进行个案调查, 同时选择同村性别相同、 年龄相近的血检阴性者作为对照, 按1∶2进行匹配, 开展病例对照研究, 以探索片形吸虫感染的危险因素。 结果 结果 共血检1 207人, 36人阳性, 人群片形吸虫血清抗体总阳性率为2.98%; 不同性别及年龄组人群血检阳性率差异均无统计学意义（χ2 = 1.686、 7.106, P均 > 0.05）; 但不同行政村及自然村人群感染率差异均有统计学意义 （χ2 = 4.103、 13.181,P 均< 0.05）。血检阳性者中有2例粪检阳性, 其中1例为新病例, 粪检阳性率为6.45% （2/31）。病例对照研究表明, 食用外购凉拌菜 （4.30）、 生薄荷 （1.22）、 生韭菜 （4.30） 和饮生水史 （3.75） OR值较高, 但均无统计学意义 （χ2 = 0.524、 0.046、 0.524、 1.571,P均 > 0.05）。 结论 结论 宾川州城地区片形吸虫感染仍然存在, 呈散发状态。进食被片形吸虫囊蚴污染的生蔬菜和生水可能是当地人群片形吸虫感染途径, 寻找到确切入口途径是控制感染的关键。     

Head and neck cancer in the betel quid chewing area: recent advances in molecular carcinogenesis

Head and neck cancer (HNC) is one of the 10 most frequent cancers worldwide, with an estimated over 500 000 new cases being diagnosed annually. The overall 5-year survival rate in patients with HNC is one of the lowest among common malignant neoplasms and has not significantly changed during the last two decades. Oral cavity squamous cell carcinoma (OSCC) shares part of HNC and has been reported to be increasing in the betel quid chewing area in recent years. During 2006, OSCC has become the sixth most common type of cancer in Taiwan, and it is also the fourth most common type of cancer among men. It follows that this type of cancer wreaks a high social and personal cost. Environmental carcinogens such as betel quid chewing, tobacco smoking and alcohol drinking have been identified as major risk factors for head and neck cancer. There is growing interest in understanding the relationship between genetic susceptibility and the prevalent environmental carcinogens for HNC prevention. Within this review, we discuss the molecular and cellular aspects of HNC carcinogenesis in Taiwan, an endemic betel quid chewing area. Knowledge of molecular carcinogenesis of HNC may provide critical clues for diagnosis, prognosis, individualization of therapy and molecular therapeutics. ( Cancer Sci 2008; 99: 1507–1514)     

Factors influencing time to smear conversion in patients with smear-positive pulmonary tuberculosis

Background and objective:   Persistent smear-positivity in patients with pulmonary tuberculosis has been shown to predict an unfavourable outcome. This study was conducted to identify the factors influencing time to sputum smear conversion.
Methods:   From July 2003 to June 2007, all patients with smear-positive and culture-confirmed pulmonary tuberculosis, who had attended a medical centre and a local teaching hospital, were identified. Factors that might have influenced time to smear conversion were investigated using time-to-event analysis.
Results:   Altogether 305 patients (mean age: 58.6 years) were studied. Diabetes mellitus was the most common underlying comorbidity. Eight patients (2.6%) had AIDS. After 2 months of treatment, 34 (11.1%) patients remained smear- and culture-positive. Cox proportional hazard regression analysis indicated that the presence of a cavity on CXR, smear grading and the first 2-month treatment regimen were independent factors influencing the time to sputum smear conversion. Among patients who had received isoniazid in the first 2 months of treatment, the time to sputum smear conversion in the 24 patients whose isolate showed isoniazid resistance was not different from that in the 236 patients whose isolate was isoniazid-susceptible (hazard ratio 1.061; 95% CI: 0.697–1.616).
Conclusions:   This analysis revealed that 11.1% of tuberculosis patients remained smear-positive after 2 months of treatment. Patients with cavitation, higher smear grading and those who had not used isoniazid, rifampicin, ethambutol and pyrazinamide continuously in the initial treatment phase had a longer time to sputum smear conversion.