炎症性疾病早期诊断的新指——sTREM-1

髓系细胞表达的触发受体-1(TREM-1)是新近确认的一个与炎症相关的免疫球蛋白超家族成员,表达于中性粒细胞、成熟的单核细胞、巨噬细胞表面.多种细菌性成分能使细胞表面的TREM-1表达增加,后者与Toll样受体协同作用,激发炎症因子的产生.最新的研究发现,感染过程中一种可溶性TREM-1(sTREM-1)可释放进入体液,并且与多种疾病如肺炎、细菌性脑膜炎、炎症性肠病密切相关,成为一种早期诊断炎症性疾病的新指标.     

N-myristoyltransferase: A potential novel diagnostic marker for colon cancer

Background  

Colon cancer is the second leading cause of cancer deaths in the western world. If detected early, colorectal cancer is one of the most treatable forms of cancer. Unfortunately, very few people are screened. N-myristoyltransferase (NMT) catalyzes myristoylation of various proteins including oncoproteins. We have demonstrated earlier the alteration of NMT activity during the progression of colorectal cancer and established NMT as a putative therapeutic target for cancer.     

Analysis of Entamoeba histolytica excretory-secretory antigen and identification of a new potential diagnostic marker

Serodiagnosis of amoebiasis remains the preferred method for diagnosis of amoebic liver abscess (ALA). However, the commercially available kits are problematic in areas of endemicity due to the persistently high background antibody titers. Human serum samples (n = 38) from patients with ALA who live in areas of endemicity were collected from Hospital Universiti Sains Malaysia during the period of 2008 to 2010. Western blots using excretory-secretory antigen (ESA) collected from axenically grown Entamoeba histolytica were probed with the above serum samples. Seven antigenic proteins of ESA with various reactivities were identified, i.e., 152 kDa, 131 kDa, 123 kDa, 110 kDa, 100 kDa, 82 kDa, and 76 kDa. However, only the 152-kDa and 110-kDa proteins showed sensitivities above 80% in the Western blot analysis. All the antigenic proteins showed undetectable cross-reactivity when probed with healthy human serum samples (n = 30) and serum samples from other infections (n = 33). From the matrix-assisted laser desorption ionization-two-stage time of flight (MALDI-TOF/TOF) analysis, the proteins were identified as heavy subunits of E. histolytica lectin and E. histolytica pyruvate phosphate dikinase, respectively. Use of the E. histolytica lectin for diagnosis of ALA has been well reported by researchers and is being used in commercialized kits. However, this is the first report on the potential use of pyruvate phosphate dikinase for diagnosis of ALA; thus, this molecule merits further evaluation on its diagnostic value using a larger panel of serum samples.     

炎症性疾病早期诊断的新指标—sTREM-1

髓系细胞表达的触发受体-1（TREM-1）是新近确认的一个与炎症相关的免疫球蛋白超家族成员，表达于中性粒细胞、成熟的单核细胞、巨噬细胞表面。多种细菌性成分能使细胞表面的TREM-1表达增加，后者与Toll样受体协同作用，激发炎症因子的产生。最新的研究发现，感染过程中一种可溶性TREM-1（sTREM-1）可释放进入体液，并且与多种疾病如肺炎、细菌性脑膜炎、炎症性肠病密切相关，成为一种早期诊断炎症性疾病的新指标。     

Retrieved endogenous biotin: a novel marker and a potential pitfall in diagnostic immunohistochemistry

 

Aim:

Antigen retrieval (AR) procedures are based on the effect of heating (by either microwave or pressure cooking treatments) on routinely fixed and paraffin embedded tissues. We observed that AR procedures restore the reactivity of endogenous biotin (EB) and report on the distribution of EB following AR in a series of routinely fixed and embedded tissues.  

Methods and results:

Following pressure cooking or microwave treatments, a simple streptavidin–peroxidase staining revealed retrieved endogenous biotin (REB) in normal tissues (such as liver, kidney and adrenal cortex), in oxyphylic cells and in some tumours, especially in carcinomas of the kidney and of the adrenal cortex. In formalin-fixed (but not in alcohol-fixed) tissue sections, the heating procedures caused an intense and finely granular cytoplasmic reaction, following a routine streptavidin-conjugated peroxidase treatment. The staining was prevented by blocking of EB by a sequential avidin–biotin treatment.  

Conclusions:

Retrieval of EB reactivity can cause pitfalls in diagnostic immunohistochemistry but, alternatively, it might also constitute a useful and novel diagnostic marker.     

Infertility: Beta-1 integrin cell adhesion molecules in the endometrium of fertile and infertile women

In order to investigate whether the endometrium of women withunexplained infertility differs immunologically from the endometriumof normal fertile women, a panel of six monoclonal antibodieswas used to characterize the presence of the ₁-integrins orvery-late-activation antigens (VLA) in the different endometrialcompartments. Precisely timed endometrial biopsies at 4, 7,10 and 13 days following the luteinizing hormone surge wereobtained from 24 normal fertile women (group I) and 24 womensuffering from unexplained infertility (group II). Frozen sectionswere labelled using an avidin-biotin peroxidase technique. VLA-1,VLA-2 and VLA-3 were present in glandular epithelium, stromalcells and vessels of both groups. VLA-4 was detected in groupI but was absent from glandular and surface epithelium of groupII. VLA-5 was not present in any of the specimens. VLA-6 wasidentified primarily in the basement membrane of vessels, glandularand surface epithelium in both patient groups. This study indicatesthat most ₁-integrins are present in endometrium throughoutthe luteal phase of the menstrual cycle. The differences observedbetween the two groups may contribute to unexplained infertility.     

Sputum microbiota as a potential diagnostic marker for multidrug-resistant tuberculosis

The prevalence of drug-resistant Mycobacterium tuberculosis (Mtb) strains makes disease control more complicated, which is the main cause of death in tuberculosis (TB) patients. Early detection and timely standard treatment are the key to current prevention and control of drug-resistant TB. In recent years, despite the continuous advancement in drug-resistant TB diagnostic technology, the needs for clinical rapid and accurate diagnosis are still not fully met. With the development of sequencing technology, the research of human microecology has been intensified. This study aims to use 16 rRNA sequencing technology to detect and analyze upper respiratory flora of TB patients with anti-TB drug sensitivity (DS, n = 55), monoresistance isoniazide (MR-INH, n = 33), monoresistance rifampin (MR-RFP, n = 12), multidrug resistance (MDR, n = 26) and polyresistance (PR, n = 39) in southern China. Potential microbial diagnostic markers for different types of TB drug resistance are searched by screening differential flora, which provides certain guiding significance for drug resistance diagnosis and clinical drug use of TB. The results showed that the pulmonary microenvironment of TB patients was more susceptible to infection by external pathogens, and the infection of different drug-resistant Mtb leads to changes in different flora. Importantly, seven novel microorganisms (Leptotrichia, Granulicatella, Campylobacter, Delfitia, Kingella, Chlamydophila, Bordetella) were identified by 16S rRNA sequencing as diagnostic markers for different drug resistance types of TB. Leptotrichia, Granulicatella, Campylobacter were potential diagnostic marker for TB patients with INH single-resistance. Delftia was a potential diagnostic marker for TB patients with RFP single drug-resistance. Kingella and Chlamydophila can be used as diagnostic markers for TB patients with PR. Bordetella can be used as a potential diagnostic marker for identification of TB patients with MDR.     

Visible and non-visible endometriosis at laparoscopy in fertile and infertile women and in patients with chronic pelvic pain: a prospective study

In 100 consecutive patients who were undergoing laparoscopyfor infertility (group 1, n = 52), chronic pelvic pain (group2, n = 18) or tubal sterilization (group 3, n = 30, asymptomaticfertile women), peritoneal biopsies were taken from areas ofvisually normal peritoneum of uterosacral ligaments. Twenty-sixpatients in group 1 (50%), eight patients in group 2 (44.4%)and 13 patients in group 3 (43.3%), were found to have laparoscopicevidence of endometriosis elsewhere in the pelvis. The majorityof women (80.7% in group 1, 87.5% in group 2, and 100% in group3) had stage I disease. The incidence of the distinctive appearancesof the lesions was similar in the three groups of patients and7% of all women or 15% (7/47) of those patients having endometriosisat laparoscopy had only subtle (non-‘typical’) endometrioticperitoneal lesions. Uterosacral biopsies showed the presenceof endometriotic tissue in three cases (5.7%), two cases (11%)and three cases (10%) in groups 1, 2, and 3 respectively. Oneof the two patients in group 2 and two of the three patientsin group 3 had no evidence of endometriosis at laparoscopy;thus histological study revealed the presence of endometriosisin normal peritoneum hi 11 % (5/47) of patients having macroscopicendometriosis and hi 6% (3/53) of patients without endometriosisat laparoscopy. Previous oral contraceptive users were significantlyhigher among women having macroscopic and/or microscopic endometriosisthan among women without the condition. In conclusion, our prospectivestudy shows a high prevalence (45–50%) of endometriosis(including microscopic forms) in both patients with chronicpelvic pain and asymptomatic women (fertile and infertile),thus supporting the modern concept that in many women endometriosismay be a paraphysiological condition while probably only hisome patients small amounts of endometriosis are an ‘annoyance’with implications to their reproductive health and may producesymptoms (eg. pelvic pain) and therefore should be defined asa ‘dis-ease’. Previous use of oral contraceptivesmay increase the risk of developing endometriosis.     

Loss of SMARCE1 expression is a specific diagnostic marker of clear cell meningioma: a comprehensive immunophenotypical and molecular analysis

Clear cell meningioma (CCM) is a rare grade II histopathological subtype that usually occurs in young patients and displays high recurrence rate. Germline SMARCE1 mutations have been described in hereditary forms of this disease and more recently in small syndromic and sporadic CCM series. The diagnostic value of SMARCE1 in distinguishing between CCM and other meningioma variants has not been yet established. The aim of our study was to investigate the status of SMARCE1 in a series of CCMs and its morphological mimickers. We compared the performance of an anti‐SMARCE1 antibody and the molecular analysis of the SMARCE1 gene in a retrospective multicenter series of CCMs. All CCMs lossed SMARCE1 immunoexpression. Bi‐allelic inactivating events were found by NGS‐based sequencing in all of these cases, except for one, which was incompletely explored, but had a wild‐type sequence. We then validated the anti‐SMARCE1 antibody specificity by analyzing additional 305 pediatric and adult meningiomas of various subtypes and 15 non‐meningioma clear cell tumors by SMARCE1 immunohistochemistry. A nuclear immunostaining was preserved in all other meningioma variants, as well as non‐meningioma clear cell tumors. In conclusion, our series showed, for the first time, that SMARCE1 immunostaining is a highly sensitive biomarker for CCM, useful as a routine diagnostic biomarker.     

Characterization of plasma inhibin forms in fertile and infertile men

BACKGROUND: The reciprocal relationship between plasma FSH and inhibin B generally reflects the state of spermatogenesis but data in some settings indicate further complexity in their relationship. Inhibin circulates as a range of higher molecular weight (mol wt) forms of varying bioactivity such that the serum profile of inhibin forms may differ between normal men and those with varying types of spermatogenic failure. The aim of this study was to establish if the inhibin B mol wt distribution was altered in men with infertility. METHODS: The mol wt profiles of inhibin B and free alpha-subunit were determined in plasma of fertile (n = 11) and infertile (n = 17) men using a combined immunoaffinity chromatography, preparative SDS-PAGE and electro-elution procedure and fractions assayed using ELISAs for inhibin B, total inhibin (all forms containing the alpha-subunit) and free alpha-subunit. RESULTS: Inhibin B was identified as precursor (60-65 k) and mature (26-30 k) forms in plasma in similar proportions (29%) in fertile men and oligospermic men (25%), but was undetectable in azoospermic men. The free alpha-subunit detected by the pro-alphaC ELISA was identified as both the precursor and processed (pro-alphaC) forms with similar proportions in fertile (8%) and all infertile (4-14%) men. The pro-alphaC ELISA did not detect the precursor forms of inhibin B in plasma while the inhibin B ELISA detected all total inhibin forms following removal of pro-containing forms by immunoabsorption. CONCLUSIONS: (i) the proportions of precursor inhibin B and alpha-subunit forms in the circulation are unchanged in men with spermatogenic disorders indicating there is no alteration of the Sertoli cell inhibin secretory pattern; (ii) these fractionation studies indicate that pro-alphaC and inhibin B ELISAs specifically detect the free alpha-subunit and inhibin B forms present in male plasma.     

Intracellular cytokine production in peripheral blood lymphocytes: a comparison of values in infertile and fertile women

Citation
Horká P, Jaro?ová R, Malí?ková K, Janatková I, Mare?ková H, Zima T, Kalousová M. Intracellular cytokine production in peripheral blood lymphocytes: a comparison of values in infertile and fertile women. Am J Reprod Immunol 2011; 65: 466–469 Problem To analyze the relation of the fertility and pregnancy of women of childbearing age to the intracellular (IC) production of tumor necrosis factor alpha (TNF‐α), interferon gamma (IFN‐γ), and interleukins 2 and 4 (IL‐2 and IL‐4). Method of study Intracellular cytokine production in peripheral blood CD3⁺ CD4⁺ lymphocytes was analyzed by flow cytometry in 185 women being treated for infertility and 50 fertile women of childbearing age. Results Infertile women have a significantly higher IC production of TNF‐α, IFN‐γ, IL‐2, and IL‐4 and higher ratios of TNF‐α/IL‐2, TNF‐α/IL‐4, and TNF‐α/IFN‐γ compared to the fertile women. Conclusion Cytokines produced by Th lymphocytes are important in orchestrating the immune response during conception, and Th‐cell dysregulation could be a reason for infertility.     

Uterus and endometrium: Immunohistochemical localization of extracellular matrix proteins in luteal phase endometrium of fertile and infertile patients

The lack of expression of certain components involved in celladhesion and migration is believed to contribute to endometrialdysfunction and implantation failure. The purpose of this studywas to investigate whether luteal phase endometrium in womenwith unexplained infertility differs, with respect to specificextracellular matrix (ECM) proteins, from endometrium of normalfertile women. A panel of monoclonal antibodies to collagentype IV, fibronectin and laminin was used to characterize thelocalization of ECM components in the different endometrialcompartments. Precisely timed endometrial biopsies obtainedat 4, 7, 10 and 13 days following the luteinizing hormone surgewere obtained from 22 normal fertile women (group 1) and 24women suffering from unexplained infertility (group 2). Paraffin-embeddedsections were labelled using the streptavidin-biotin alkalinephosphatase technique. In group 1, collagen type IV, fibronectinand laminin were absent from the luminal epithelium but presentin stromal cells and the basement membrane of glands and bloodvessels. In group 2, these components were absent from all endometrialregions using equivalent titres of antibody to those used ingroup 1. This suggests that the endometrium of women with unexplainedinfertility demonstrates defects in the distribution of certainECM glycoproteins. A possible consequence of this defect maybe implantation failure.     

Nuclear beta catenin as a potential prognostic and diagnostic marker in patients with colorectal cancer from Hong Kong.

AIMS: To study the expression of nuclear beta catenin in patients with colorectal cancer, colorectal adenoma, and colorectal polyps to elucidate its role in carcinogenesis, and its potential for prognosis and diagnosis. METHODS: The expression of nuclear beta catenin was studied by immunohistochemistry using paraffin wax embedded specimens. Sixty specimens each of colorectal carcinoma, colorectal adenoma, colorectal polyp, and normal colorectal specimens were analysed. The potential for prognosis was assessed by correlating nuclear beta catenin expression in 60 and 75 patients with colorectal cancer with lymph node metastasis and survival, respectively. The diagnostic capacity was explored by comparing nuclear beta catenin expression in 60 patients with colorectal cancer with other cytokeratin 20 (CK20) positive adenocarcinomas, namely: 30 colonic mucinous adenocarcinomas, 30 gastric adenocarcinomas, 27 pancreatic adenocarcinomas, and 12 ovarian mucinous adenocarcinomas. RESULTS: Nuclear beta catenin expression was highly associated with progression of colorectal tissue from normal epithelial tissue, polyps, adenomas, to carcinomas (r = 0.875; p < 0.0001). Nineteen patients with colorectal adenoma who subsequently developed colorectal carcinoma had higher nuclear beta catenin expression than those with colorectal adenomas alone (p < 0.0001). Moreover, those patients with colorectal cancer and high nuclear beta catenin expression had a higher incidence of lymph node metastasis (chi(2) = 16.99; p < 0.005) and shorter overall survival (p < 0.0001). Finally, nuclear beta catenin expression in colorectal adenocarcinomas was significantly higher than in other CK20 positive adenocarcinomas. CONCLUSIONS: Nuclear beta catenin expression is a potential prognostic factor in patients with colorectal cancer, and together with CK20, it could be used to identify colorectal carcinoma in the Hong Kong population.     

Podoplanin: a novel diagnostic immunohistochemical marker

Podoplanin is a transmembrane mucoprotein recognized by the recently commercially available D2-40 monoclonal antibody. Recent investigations have shown that podoplanin is selectively expressed in lymphatic endothelium as well as lymphangiomas, Kaposi sarcomas, and in a subset of angiosarcomas with probable lymphatic differentiation. Podoplanin has also been shown to be strongly expressed in seminomas, epithelioid mesotheliomas, and hemangioblastomas, and immunostaining for this marker can assist in the diagnosis of these tumors. This article reviews the current information on the applications of podoplanin immunostaining in surgical pathology.     

Monoclonal antibody to desmosomal glycoprotein 1--a new epithelial marker for diagnostic pathology

Desmosomes are intercellular adhesive junctions that occur in almost all epithelia and should therefore be useful as epithelial markers in tumour diagnosis. Here, we describe a monoclonal antibody, 32-2B, to a major desmosomal glycoprotein (dgl) which reacts with human tissues in paraffin sections. This antibody was tested for its ability to stain epithelia and tumours. It reacted with all epithelia tested and with every specimen of a wide range of carcinomas. It also stained meningiomas, another desmosome-containing tumour. It did not stain other types of tumours including lymphomas, melanomas, and various sarcomas, or normal tissues which lack desmosomes. These characteristics demonstrate that 32-2B is a reliable epithelial marker that may have a useful role in diagnostic histopathology.