Use of the duplex TaqMan PCR system for detection of Shiga-like toxin-producing Escherichia coli O157

Real-time PCR assays have been applied for the detection and quantification of pathogens in recent years. In this study two combinations of primers and fluorescent probes were designed according to the sequences of the rfb(Escherichia coli O157) and stx2 genes. Analysis of 217 bacterial strains demonstrated that the duplex real-time PCR assay successfully distinguished the Escherichia coli O157 serotype from non-E. coli O157 serotypes and that it provided an accurate means of profiling the genes encoding O antigen and Shiga-like toxin 2. On the other hand, bacterial strains that lacked these genes were not detected by this assay. The quantitative ranges of the real-time PCR assay for these two genes were linear for DNA concentrations ranging from 10(3) to 10(9) CFU/ml of E. coli O157:H7 in pure culture and milk samples. The real-time PCR allowed the construction of standard curves that facilitated the quantification of E. coli O157:H7 in feces and apple juice samples. The detection sensitivity of the real-time PCR assay ranged from 10(4) to 10(9) CFU/g (or 10(4) to 10(9) CFU/ml) for feces and apple juice and 10(5) to 10(9) CFU/g for the beef sample without enrichment. After enrichment of the food samples in a modified tryptic soy broth, the detection range was from 10(0) to 10(3) CFU/ml. The real-time PCR assays for rfb(E. coli) (O157) and stx2 proved to be rapid tests for the detection of E. coli O157 in food matrices and could also be used for the quantification of E. coli O157 in foods or fecal samples.     

Cryptococcal antigen test revisited: significance for cryptococcal meningitis therapy monitoring in a tertiary chinese hospital

For a total of 29 non-human immunodeficiency virus 1 cryptococcal meningitis cases, titer changes in the latex agglutination test before and after therapy were reviewed along with clinical manifestations, laboratory findings, and therapy regimens. The cryptococcal antigen titer decreased for every case after therapy and was correlated to fungal clearance as defined by fungus smear and/or culture. However, cryptococcal antigen can remain at low titers for long periods of time after therapy, even when fungus smears and/or cultures become negative.     

Expression of PKC-beta or cyclin D2 predicts for inferior survival in diffuse large B-cell lymphoma.

We sought to determine whether identification of poor-risk subgroups of diffuse large B-cell lymphoma (DLBCL) using immunohistochemical stains would have practical utility with regard to prognosis and therapeutic decisions. Tissue microarray blocks were created using replicate samples of formalin-fixed, paraffin-embedded tissue from 200 cases of de novo DLBCL. The sections were stained with antibodies to proteins that are expressed by activated or proliferating B cells including MUM1, FOXP1, bcl-2, survivin, protein kinase C-beta (PKC-beta), cyclin D2, cyclin D3, and Ki-67. In univariate analysis, tumor expression of cyclin D2 (P = 0.025) or PKC-beta (P = 0.015) was associated with a worse overall survival, whereas none of the other markers was predictive of overall survival. Patients with DLBCL that expressed either cyclin D2 or PKC-beta had a 5-year overall survival of only 30% as compared to 52% for those who were negative for both markers (P = 0.0019). In multivariate analysis, the expression of cyclin D2 or PKC-beta was an independent predictor of poor overall survival (P = 0.035). Cyclin D2 and PKC-beta expression will be useful in designing a 'biological prognostic index' for patients with DLBCL.     

Dose reduction for the management of indinavir-related toxicity in human immunodeficiency virus type 1-infected patients in Taiwan: clinical and pharmacokinetic assessment.

This study evaluated the feasibility of reducing the indinavir (IDV) dosage in Taiwanese patients receiving the standard IDV/ritonavir (RTV) dosage of 800/100 mg twice a day who had undetectable plasma human immunodeficiency virus type 1 (HIV-1) RNA but had developed IDV-related toxicities. After dosage reduction to IDV/RTV 600/100 mg twice a day, the dose-related toxicity decreased and plasma HIV RNA remained undetectable at 24 weeks post-switch in all patients. The maximal plasma concentration (Cmax) and area under the plasma concentration-time curve of IDV decreased significantly (median, 6.3 vs 4.3 microg/mL and 1892 vs 1292 microg.min/mL, p=0.01 and 0.001, respectively) but the minimal plasma concentration remained at a similar level (median, 1.0 vs 0.8 microg/mL, p=0.12). This study found that the reduction in the dosage of IDV in HIV-1 infected patients receiving the standard IDV/RTV regimen guided by therapeutic drug monitoring decreased the Cmax, dose-related toxicity and medical cost without compromising viral control.     

Characterization of <Emphasis Type="Italic">p</Emphasis>24 Gene of <Emphasis Type="Italic">Spodoptera litura</Emphasis> Multicapsid Nucleopolyhedrovirus

Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) p24 gene is 753 bp long, potentially encoding 244 amino acids with a predicted molecular weight of 27.3 kDa. Homology analysis indicated that SpltMNPV P24 has 20–36% amino acid identity with that of other known baculoviruses. RT-PCR results showed that the p24 gene is transcribed actively at the late stage of infection and the mRNA start site was mapped within a consensus baculovirus late promoter sequence (ATAAG). Western blot analysis of extracts from SpltMNPV-infected S. litura cells detected a specific 28 kDa protein, and this protein was not N-glycosylated. Structural localization revealed that SpltMNPV P24 was associated with the nucleocapsid of occlusion-derived virus (ODV) as a complex form of 83 kDa.     

早老素1基因在转染CHO细胞中的表达及其与γ-分泌酶的关系

目的 研究早老素1(PS1)在淀粉样前体蛋白(APP)加工生成β-淀粉样多肽(Ap)过程中的作用及其与γ-分泌酶的关系。方法构建APP和PSI双基因稳定转染的中国仓鼠卵巢(CHO)细胞株,应用免疫沉淀和印迹、脉冲追踪及ELISA方法,检测PS1的表达和代谢半衰期,分析对Aβ分泌的影响及与γ-分泌酶功能的关系。结果PS1转染的CHO细胞(APP-PSI)表达的主要是相对分子质量为45000的全长PS1蛋白,其半衰期短于1h,而其活性片段的N-末端片段和C-末端片段则相对稳定,半衰期接近16h。突变型PS1(M146L)转染细胞分泌的Ap总量与野生型PS1转染细胞没有明显差别,但分泌的Aβ亚型Ap142是未转染PS1或野生型PS1转染细胞分泌的将近2倍。结论 PS1参与了APP加工生成Aβ的过程,突变型PS1(M16L)导致Aβ142的分泌增加,提示PS1可能就是预期的γ-分泌酶。     

伴玻璃样间质的透明细胞癌二例

例1男,76岁。因双下肢浮肿2个月余,声嘶1周于2000年3月12日入院。体检:一般情况差,左颈部触及肿大淋巴结约2cm×2cm大小,血生化检查提示肾功能及呼吸功能衰竭,后因呼吸困难作气管插管时舌根部见一菜花状肿瘤约3.0cm×2.5cm大小,行颈淋巴结及舌根部肿瘤活检术。术后1个月患者死于慢性肾炎所致肾功能及呼吸功能衰竭。病理检查:切除淋巴结大小2cm×2cm,切面灰白色,质硬。舌根部肿瘤大小3.0cm×2.5cm,表面呈菜花状,切面灰白色,质硬。镜下观察:舌根部肿瘤细胞形态较一致,排列呈互相吻合的小梁状、岛状或片状。巢间有大量玻璃样变的纤维性间质。瘤…     

免疫组织化学和电镜在腹膜间皮瘤和浆液性癌鉴别诊断中的价值

上皮样间皮瘤，特别是那些呈乳头状和管状乳头状生长的间皮瘤，与原发性或转移性浆液性乳头状癌弥漫累及腹膜者很难鉴别。1998年一个研究报道有16种标记在间皮瘤的诊断中有一定的价值，该项研究认为，calretinin、thrombomodulin和CK5／6是间皮瘤最好的诊断指标，MOC-31、B72．3、Ber—EP4、CA19—9和Leu—M1（CD15）在间皮瘤中表达阴性，可用于区分腹膜上皮样恶性间皮瘤和乳头状浆液性癌弥漫累及腹膜。D2-40和podoplanin是2个最近认识的淋巴管内皮细胞标记物，它们也表达于正常间皮细胞和间皮瘤。为了比较这两种新的间皮细胞标记和以前其他用于间皮瘤和浆液性癌鉴别诊断的标记的特异性，     

子宫癌和子宫肌瘤患者血清PRL、T和E2含量的变化

目的：探讨人血清催乳素（PRL）、睾酮（T）和雌二醇（E2）与子宫肌瘤、子宫癌的关系。方法：采用化学发光免疫法（CLIA）分别检测了子宫肌瘤患者（30例）、子宫癌患者（28例）的血清PRL、T和E2三类激素并与健康人组（38人）比较。结果：1．子宫肌瘤和子宫癌的PRL和T含量均明显高于正常组（P〈0．01）。2．子宫肌瘤与子宫癌组的E2水平与正常人无明显差异（P〉0．01）。3．从正常人组经子宫肌瘤患者到子宫癌组其血PRL水平呈持续增高趋势。结论：PRL和T含量的变化与子宫肌瘤及子宫癌疾患有密切关系。     

Michael Addition Polymerizations of Trifunctional Amines with Divinyl Sulfone

Summary: The mechanisms of the Michael addition polymerization of N‐aminoethyl piperazine (AEPZ) with divinyl sulfone (DVS) were clarified based on the reactivity sequence of three different amines in AEPZ: 2° amine in piperazine ring > 1° amine ≫ 2° amine formed in situ. When the feed molar ratio of DVS to AEPZ was 1:1, the polymerization of AB intermediate formed proceeded, and the linear poly(sulfone amine) containing secondary and tertiary amines in the backbones were produced. The linear structure of the product was confirmed by NMR spectra, and the molecular weights, molecular weight distribution, and properties of poly(sulfone amine)s were characterized by GPC, DSC, and TGA.

Polymerization of linear poly(DVS‐AEPZ).