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Background T cell immunoglobulin mucin-3 (Tim-3) has been reported to participate in the regulation of immune response and the induction of allograft tolerance. However, the association between Tim-3 and renal allograft dysfunction is unclear. We studied the expression of cellular and soluble Tim-3 (sTim-3), soluble galectin-9 (sGal-9) and carcinoembryonic antigen-related cell adhesion molecule-1 (sCEACAM-1) in kidney transplantation recipients (KTRs) to explore their roles in allograft dysfunction. Methods 96 KTRs (53 with stable graft and 43 with graft dysfunction) and 30 healthy controls (HC) were enrolled. Among the KTRs, 55 used Tacrolimus (TAC) and 41 used Sirolimus (SRL). In the dysfunction group, 29 recipients have undergone graft biopsy and 14 were classified as biopsy-proven rejection (BPR). Cellular Tim-3 was determined by flow cytometry. sTim-3 was determined by ELISA. sGal-9 and sCEACAM-1 were determined by Bio-Plex® suspension array system. Results KTRs with renal dysfunction showed significantly higher levels of sTim-3 and sGal-9 but similar levels of cellular Tim-3 and sCEACAM-1 compared with stable recipients. Correlation analysis revealed that estimated glomerular filtration rate (eGFR) was negatively associated with sTim-3 and sGal-9. Both BPR and non-BPR groups showed comparable levels of Tim-3, Gal-9 and CEACAM-1. Moreover, SRL group showed significantly higher levels of sCEACAM-1 than TAC and HC groups. Conclusions sTim-3 and sGal-9 were promising biomarkers for allograft dysfunction, but unable to differentiate allograft rejection from other causes of renal dysfunction in KTRs. Moreover, long-term administration of sirolimus would up-regulate sCEACAM-1 level, while exert similar regulatory effects on Tim-3 and Gal-9 compared to tacrolimus.

Circulating tumour cell (CTC) enumeration and profiling has been established as a valuable clinical tool in many solid malignancies. A key challenge in CTC research is the limited number of cells available for study. Ex vivo CTC culture permits expansion of these rare cell populations for detailed characterisation, functional assays including drug sensitivity testing, and investigation of the pathobiology of metastases. We report for the first time the establishment and characterisation of two continuous CTC lines from patients with gastroesophageal cancer. The two cell lines (designated UWG01CTC and UWG02CTC) demonstrated rapid tumorigenic growth in immunodeficient mice and exhibit distinct genotypic and phenotypic profiles which are consistent with the tumours of origin. UWG02CTC exhibits an EpCAM+, cytokeratin+, CD44+ phenotype, while UWG01CTC, which was derived from a patient with metastatic neuroendocrine cancer, displays an EpCAM−, weak cytokeratin phenotype, with strong expression of neuroendocrine markers. Further, the two cell lines show distinct differences in drug and radiation sensitivity which match differential cancer-associated gene expression pathways. This is strong evidence implicating EpCAM negative CTCs in metastasis. These novel, well characterised, long-term CTC cell lines from gastroesophageal cancer will facilitate ongoing research into metastasis and the discovery of therapeutic targets.

As noble metal nanoclusters (NCs) are widely employed in nanotechnology, their potential threats to human and environment are relatively less understood. Herein, the biological effects of ultra-small silver NCs coated by bovine serum albumin (BSA) (Ag-BSA NCs) on isolated rat liver mitochondria were investigated by testing mitochondrial swelling, membrane permeability, ROS generation, lipid peroxidation and respiration. It was found that Ag-BSA NCs induced mitochondrial dysfunction via synergistic effects of two different ways: (1) inducing mitochondrial membrane permeability transition (MPT) by interacting with the phospholipid bilayer of the mitochondrial membrane (not with specific MPT pore proteins); (2) damaging mitochondrial respiration by the generation of reactive oxygen species (ROS). As far as we know, this is the first report on the biological effects of ultra-small size nanoparticles (∼2 nm) at the sub-cellular level, which provides significant insights into the potential risks brought by the applications of NCs. It would inspire us to evaluate the potential threats of nanomaterials more comprehensively, even though they showed no obvious toxicity to cells or in vivo animal models. Noteworthy, a distinct toxic mechanism to mitochondria caused by Ag-BSA NCs was proposed and elucidated.

This study seeks to compare the impact of selective partial portal vein ligation (PPVL) or the combination of simultaneous hepatic artery ligation (PPVAL) with in situ splitting (ISS) on liver regeneration and injury. Rats were randomized into three groups; namely: selective PVL, PPVL + ISS and PPVAL + ISS. The changes in hepatic hemodynamics, liver regeneration and hepatocytic injury were examined. Blood flow to the left portal branch and the microcirculation of the left median lobe after PPVL or PPVAL was significantly reduced. Liver regeneration of PPVAL + ISS group was more pronounced than that in the PPVL + ISS and PVL groups at 48 and 72 hours as well as 7 d postoperatively. The serum biochemical markers and histopathological examination demonstrated reduced levels of liver injury in the PPVL + ISS group. Injury to hepatocytes was more pronounced with PPVAL + ISS than PVL. HGF, TNF-α and IL-6 expression in the regenerated lobes in both PPVAL + ISS and PPVL + ISS groups increased significantly when compared to the PVL group. We demonstrated that both PPVL + ISS and PPVAL + ISS were effective and feasible means of inducing remnant liver hypertrophy and could serve as a rapid clinical application for qualified patients.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors with an increasing incidence worldwide. Apolipoprotein M (apoM) is a novel apolipoprotein that is mainly expressed in liver and kidney tissues. However, the anti-tumor properties of apoM remain largely unknown. We evaluated the anti-tumor activities and mechanisms of apoM in HCC both in vivo and in vitro. Bioinformatic analysis and luciferase reporter assay results showed that apoM was a potential target of hsa-miR-573 and was downregulated after transfection with hsa-miR-573 mimics. Overexpression of apoM suppressed migration, invasion, and proliferation of hepatoma cells in vitro. Overexpression of hsa-miR-573 in hepatoma cells reduced apoM expression, leading to promotion of the invasion, migration, and proliferation of hepatoma cells in vitro. In addition, hsa-miR-573 markedly promoted growth of xenograft tumors in nude mice with an accompanying reduction in cell apoptosis. ApoM markedly inhibited growth of xenograft tumors in nude mice and promoted cell apoptosis. Moreover, Bcl2A1 mRNA and protein levels were inhibited by apoM overexpression and an increase in apoptosis rate by apoM was markedly compensated by Bcl2A1 overexpression in HepG2 cells. These results provide evidence that hsa-miR-573 promoted tumor growth by inhibition of hepatocyte apoptosis and this pro-tumor effect might be mediated through Bcl2A1 in an apoM-dependent manner. Therefore, our findings may be useful to improve understanding of the critical effects of hsa-miR-573 and apoM in HCC pathogenesis.

Infection remains a major cause of morbidity and mortality after kidney transplantation (KT). Reliable biomarkers to predict post-transplant infection are lacking. We investigated the predictive performance of pre- and post-transplant levels of T-cell immunoglobulin and mucin domain-3 (Tim-3) and Galectin-9 (Gal-9), two pleiotropic immunomodulatory molecules, in early identification of infection. Serum Tim-3 and Gal-9 were paired measured before and 30 days after transplantation (PTD 30) in 95 KT recipients (KTRs). The decline rates of Tim-3 and Gal-9 were calculated relative to pre-transplant levels. KTRs with infection history had significantly higher levels of PTD 30 Tim-3 and Gal-9, and slower decrease rates of Gal-9 compared to non-infected recipients, while no difference was observed between two groups regarding pre-transplant levels. The AUCs for predicting 1-year post-transplant infection were 0.653 and 0.711 for post-transplant Tim-3 and Gal-9, 0.664 and 0.670 for relative Tim-3 and Gal-9, respectively. After adjusting for potential confounders, PTD 30 Tim-3, Gal-9 and relative Gal-9 remained as independent risk factors for post-transplant infection. Our results suggested that PTD 30 Tim-3 and Gal-9 and relative decrease of Gal-9 were promising predictors for identifying KTRs with high risk of infection, while pre-transplant Tim-3 and Gal-9 showed no predictive power to infection.

Propofol (2,6-diisopropylphenol) is probably the most widely used intravenous hypnotic agent in daily practice. However, its anti-inflammatory properties have seldom been addressed. In this study, we evaluated the anti-inflammatory activity and mechanisms of propofol on lipopolysaccharide (LPS)-induced inflammation in vivo and in vitro and found that propofol markedly inhibited LPS-induced production of pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, and expression of inducible nitric oxide synthase (iNOS). At the same time, the expression of hepatocyte nuclear factor-1α (HNF-1α) and apolipoprotein M (APOM) was inhibited by treatment with LPS and LPS-induced down-regulation of HNF-1α expression and APOM expression could be compensated by propofol treatment. However, propofol could not compensate LPS-induced down-regulation of APOM expression by treatment with HNF-1α siRNA and the suppressive effect on LPS-induced pro-inflammatory cytokines production by propofol was significantly compensated by treatment with APOM siRNA. These results provide evidence that propofol may first up-regulate APOM expression by enhancing HNF-1α expression and then inhibit pro-inflammatory cytokine production in LPS-stimulated cells. Therefore, our study may be useful in understanding the critical effect of propofol in patients with systemic inflammatory response syndrome.

Silver materials have been widely used as antimicrobial agents. Notably, silver nanoparticles have emerged as a new generation of nanoproducts for biomedical and environmental applications in recent years. However, ultrasmall silver nanoclusters (NCs) (∼2 nm) have rarely been used to kill bacteria and their antibacterial mechanisms have not yet been fully elucidated. Herein, we studied the antibacterial activities of bifunctional fluorescent DHLA-AgNCs against three types of bacteria. The results showed that DHLA-AgNCs exhibited excellent antibacterial activities against Gram-negative E. coli, which could efficiently inhibit the growth of E. coli DH 5α and E. coli DSM 4230 cells at a concentration of 15 and 10 μg mL-1, respectively. Meanwhile AgNCs demonstrated no apparent antibacterial activity against Gram-positive S. aureus. Then, the antibacterial mechanisms of AgNCs were systematically investigated. We found that AgNCs affected the growth of different E. coli strains in different ways. AgNCs inhibited the growth of E. coli DH 5α mainly through damaging the outer cellular membrane and permeating into the cells, followed by the antibacterial effect of the internalized AgNCs and released silver ions. AgNCs, however, inhibited the growth of E. coli DSM 4230 cells mainly through diffusing into E. coli DSM 4230 cells and damaging their respiratory chain. These results clearly indicated that different bacterial strains (e.g. different E. coli strains) should be taken into consideration in future studies. Our work facilitates further investigation of the design of new antibacterial silver nanomaterials with different sizes.

Aim IgG4 disease is rare. However, IgG4 tubulointerstitial nephritis (TIN) is the most common renal manifestation. IgG4 disease is usually associated with elevated serum IgG4 levels and other organ involvement, low-density renal lesions on enhanced CT imaging and immune activation. The incidence of IgG4-TIN may be underestimated, as staining for IgG4 is not routine. This study sought to describe the prevalence of previously undiagnosed IgG4-TIN. Due to the complexity of the diagnosis, we only attempt to look at IgG4-positive plasma cell TIN as a potential indication for IgG4 renal disease. Methods A retrospective review of native renal biopsies performed between 2002 and 2012 with a primary diagnosis of TIN was selected. Samples for which interstitial nephritis was secondary to a glomerular disease were excluded. The tissues were stained for IgG4 and scored by two blinded observers. Demographic and follow-up details were collected. This study was approved by the local ethics committee. Results 82 cases of interstitial nephritis from a total of 1238 renal biopsies (2002–2012) were available after staining for further assessment. 12 samples demonstrated staining consistent with the criteria for IgG4-positive plasma cell TIN, of which 3 had mildly positive staining, 7 moderately positive staining and 2 had markedly positive staining. There were no statistically significant differences in the baseline characteristics between the positive and negative staining groups. Conclusions A number of cases of IgG4-positive plasma cell TIN were observed histologically that had been previously diagnosed as non-specific chronic TIN. IgG4-positive plasma cell TIN made up 1% of all renal biopsies performed over 10 years and 13% of all biopsies demonstrating TIN not related to glomerular disease. IgG4 staining should be considered routinely in biopsies demonstrating primary TIN.

Ischemia-modified albumin (IMA) levels have been advocated as a biomarker for evaluating the oxidative stress status. No data are showed on the potential role of IMA in type 1 diabetes (T1D). We aimed to establish the correlation among serum levels of IMA, C-reactive protein (CRP), and diabetic ketoacidosis (DKA) in patients with T1D. Fifty-seven patients with T1D, 27 patients with DKA, and 40 controls were enrolled. Serum IMA and CRP levels were measured and evaluated to distinguish from DKA. CRP and IMA levels were significantly elevated in patients with DKA at admission to the hospital compared to non-DKA and control subjects. CRP and IMA levels were higher in non-DKA patients than in controls. CRP, plasma glucose, and IMA levels were reduced after insulin treatment. Serum IMA levels were an independent risk marker for DKA (OR = 1.225, p = 0.002, 95 % CI: 1.076-1.394). Receiver operating characteristic curve analyses showed no difference in the areas under curve for serum IMA and CRP values. This study indicates that IMA and CRP levels were significantly correlated with DKA diagnosis. IMA can act as a biomarker that reflects the presence of DKA.

To investigate whether ischemia-modified albumin (IMA) is reliable for early diagnosing symptomatic lacunar infarction (SLI) in type 2 diabetics.Ninety-seven consecutive diabetic patients, 47 with SLI, and 45 healthy controls were enrolled. Serum IMA and plasma total homocysteine (tHcy) were measured on an autoanalyzer and evaluated in distinguishing SLI.Serum IMA levels were 97.35 ± 5.25 U/L in the healthy control group, 103.26 ± 7.43 U/L in the non-SLI group, and 139.84 ± 20.00 U/L in the SLI group. Plasma tHcy levels were 8.08 ± 1.82 μmol/L, 11.31 ± 3.03 μmol/L, and 13.10 ± 3.67 μmol/L, respectively. The differences in IMA and tHcy levels were statistically significant for all groups (p0.05). Receiver operating characteristic curve analyses revealed the areas under curve were 0.866 for IMA and 0.352 for tHcy.This study indicates that IMA was significantly elevated in the acute phase of SLI and more sensitive than tHcy in distinguishing SLI.

Two new acetylated kaempferol glycosides were isolated from the seeds of Camellia semiserrata Chi, their structures were elucidated as kaempferol 3- O -[(3- O -acetyl)-α- l -rhamnopyranosyl(1 → 3)(4- O -acetyl)- α - l -rhamnopyranosyl(1 → 6)- β - d -gluco-pyranoside] ( 1 ) and kaempferol 3- O -[(2- O -acetyl)- α - l -rhamnopyranosyl (1 → 3)(4- O -acetyl)- α - l -rhamnopyranosyl(1 → 6)- β - d -gluco-pyranoside] ( 2 ) by spectral experiments (including ESI-MS, 1D- and 2D-NMR).

BACKGROUND: The presence of glutathione S-transferase (GST) π1 (GSTP1) or multidrug resistance gene 1 (MDR1) promoter methylation in lung cancer was studied for the first time to the authors' knowledge; and, to date, the clinical significance of methylation is not clear. The objective of the current study was to determine the promoter methylation status of GSTP1 and MDR1, which encode GST-π and P-glycoprotein (Pgp), respectively, in patients with bronchioloalveolar carcinoma (BAC) and to investigate whether methyltransferase 1 (DNMT1)-mediated GSTP1 or MDR1 methylation are responsible for disease progression and prognosis in patients with BAC. METHODS: Protein expression levels of DNTM1, GST-π, and Pgp were determined by immunohistochemistry in samples from 36 patients with BAC. Promoter methylation status of the GSTP1 and MDR1 genes was determined by using methylation-specific polymerase chain reaction analysis. RESULTS: The results demonstrated a significant correlation between the methylation of the GSTP1 or MDR1 promoters and negative expression of their respective proteins in BAC (P < .05). A significant correlation also was demonstrated between GSTP1 methylation and recurrence-free and overall survival of patients with BAC. DNMT1 protein expression levels were correlated with GSTP1 promoter methylation and patient prognosis (P < .05). However, no correlation was observed between DNMT1 expression and MDR1 methylation. CONCLUSIONS: GSTP1 promoter methylation mediated by DNMT1 may promote BAC progression and could serve as a poor prognostic indicator for patients with this disease. DNMT1 protein expression also may be considered as a prognostic indicator. Methylation of the MDR1 promoter may be mediated through pathways other than DNMT1 in BAC and does not appear to be associated with disease progression or patient prognosis. Cancer 2009. © 2009 American Cancer Society.

Two new flavonol glycosides and three known flavonoids were isolated from seeds of Camellia semiserrata Chi. The structures of these new flavonol glycosides were established as kaempferol 3-O-[(2'''''',3'''''',4''''''-triacetyl)-alpha-L-rhamnopyranosyl(1--3)(2'''',4''''-diacetyl)-alpha-L-rhamnopyranosyl (1--6)-beta-D-glucopyranoside] and kaempferol 3-O-[(3'''''',4''''''-diacetyl)-alpha-L-rhamnopyranosyl(1--3)(2'''',4''''-diacetyl)-alpha-L-rhamnopyranosyl(1--6)-beta-D-glucopyranoside] by spectroscopic methods. The estrogenic activity of these compounds was investigated by a recombinant yeast screening assay.

To develop the proper renal targeting carrier for clinical therapy, human serum albumin, as starting material, was firstly cleaved into albumin fragments and Superdex 75 and CM-Sepharose FF were used to separate and purify the degradation products. Consequently, three peptide fragments (PFs) with certain sequence named PF-A 1–123 , PF-A 124–298 and PF-A 299–585 were obtained as candidates of renal targeting carrier. Then, cytotoxicity and cellular uptake of three PFs was studied preliminarily. The results showed that three PFs had no adverse effects on the HeLa and MDCK cell even up to 5.00 mg/mL and PF-A 299–585 exhibited highest affinity to MDCK cells. After that, we found that PFs selectively accumulated in the kidneys, especially in the renal tubules after intravenous injection in mice by optical imaging study. Finally, Tissues distribution in vivo was utilized to verify the renal targeting profiles of PFs. Three PFs exhibited renal accumulation characteristics. In particular, about 40% injected doses of PF-A 299–585 were specifically distributed into kidneys for 1 h. The mean area under the curve (AUC) of PF-A 299–585 in kidneys increased 13 times compared with those of PF-A 1–123 and PF-A 124–298 . Therefore, PFs can be applied as prospective carriers for renal targeting and PF-A 299–585 may be the optimal carrier.

OBJECTIVES: To investigate thyroid peroxidase gene (TPO) mutations in a Chinese siblings with congenital goitrous hypothyroidism (CGH). SUBJECTS AND METHODS: The proband, his sister, and their parents were enrolled. All subjects underwent clinical examination and laboratory tests. Mutation screening of the TPO gene was performed by sequencing fragments amplified from extracted genomic DNA. RESULTS: The siblings were diagnosed as CGH with neurodevelopmental deficits. Two compound heterozygous inactivating mutations were found in the two patients: a frameshift mutation between positions 2268 and 2269 (c.2268-2269 insT) and a missense mutation at c.2089 G>A (p.G667S) of the TPO gene. Their parents, with normal thyroid hormone levels, were heterozygous for mutations c.2268-2269 insT and c.2089 G>A, respectively. The polymorphisms of c.1207 G>T, c.1283 G>C, and c.2088 C>T were detected in the family. CONCLUSIONS: CGH of the Chinese siblings was due to the TPO gene mutations (c.2268-2269 insT and c.2089 G>A). Arq Bras Endocrinol Metab. 2012;56(9):614-7 OBJETIVOS: Investigar mutações no gene da peroxidase da tireoide (TPO) em irmãos chineses com hipotireoidismo congênito com bócio (HCB). SUJEITOS E MÉTODOS: O probando, sua irmão e seus pais foram analisados. Todos os sujeitos passaram por exames clínicos e laboratoriais. A análise para mutações do gene TPO foi feita por meio de sequenciamento de fragmentos amplificados do DNA genômico extraído. RESULTADOS: Os irmãos foram diagnosticados com HCB e déficits de desenvolvimento neurológico. Duas mutações compostas, heterozigotas, inativadoras foram observadas nos dois pacientes: uma mutação frameshift entre as posições 2268 e 2269 (c.2268-2269 insT), e uma mutação missense em c.2089 G>A (p.G667S) do gene TPO. Os pais apresentaram níveis normais de hormônios da tiroide e eram heterozigotos para mutações em c.2268-2269 insT e c.2089 G>A, respectivamente. Foram detectados polimorfismos de c.1207 G>T, c.1283 G>C, e c.2088 C>T na família. CONCLUSÕES: O HCB dos irmãos chineses foi devido a mutações no gene TPO (c.2268-2269 insT e c.2089 G>A). Arq Bras Endocrinol Metab. 2012;56(9):614-7

Multidrug resistance (MDR) to chemotherapeutic agents is a major obstacle to successful treatment in patients with breast cancer. Besides overexpression of MDR1, the member of the ABC transporter family, glucosylceramide synthase (GCS) which allows cellular escape from ceramide-induced cellular apoptosis by mediating ceramide glycosylation was considered to be related with multidrug resistance (MDR) in breast cancer. To specifically and efficiently reverse MDR of breast carcinoma cells, two small interference RNA (siRNA) targeted multidrug resistance 1 (MDR1) and GCS mediated by plasmids were constructed and co-transfected into the MDR breast cancer cell line MCF-7/ADM. The results showed that both transient transfection and stable transfection of the two siRNA-expression plasmids could greatly decrease the relative expression of GCS mRNA and MDR1 mRNA, significantly lower than the controls (p0.01). Evaluation of chemosensitivity displayed a 96-fold reduction in drug resistance for Adriamycin. And the reversing effects could keep for a long period (at least 3 w). Our results demonstrated that co-inhibition of MDR1 and GCS could more effectively reverse MDR in drug resistant breast carcinoma cells in vitro.

Summary Aims Using archival material, we studied the immunoreactivity and utility of monoclonal anti-human inhibin α subunit in the identification of chorionic villi (CV) and trophoblastic subpopulations in endometrial curettings (EC) from patients who had intra-uterine, ectopic, molar and, particularly, probable intra-uterine pregnancies. We also compared its expression with those of bHCG, HPL and CAM 5.2. Methods The four groups of EC investigated included: Group 1, 15 patients with intra-uterine pregnancies (IUP); Group 2, 15 patients with tubal pregnancies (TP); Group 3, 15 patients with hydatidiform moles (HM); and Group 4, 20 patients with purported history of intra-uterine pregnancies (PIUP). Positive and negative control cases were from Groups 1 and 3 and Group 2, respectively. The test cases were from Group 4. Immunohistochemistry was performed on each case testing for expression of inhibin α, βHCG, HPL and CAM 5.2. Results Trophoblastic populations, which included syncy-tiotrophoblast (ST), cytotrophoblast (CT) and intermediate trophoblast (IT), were absent in all 15 negative control cases (Group 2). The 30 positive control cases (Groups 1 and 3) revealed the following: (a) ST, CT and IT were identified in all cases and were positive for CAM 5.2, (b) inhibin α, βHCG and HPL (except one case) were reactive for all cases with ST, but not CT, and (c) IT positivity for βHCG, HPL and inhibin α was 67, 80–93 and 100%, respectively. From the 20 test cases (Group 4), the findings were: (a) CT was absent in all cases, (b) scattered ST cells, which were identified only in 10 cases, were positive for all antibodies, (c) scattered IT cells were present in 17 cases and showed 100% CAM 5.2 positivity, and (d) IT positivity for βHCG, inhibin α and HPL was 58.8% (10/17), 76.5% (13/17) and 82.4% (14/17), respectively. Background staining was observed in 22 of 65 cases (33.8%) stained with βHCG and HPL; half of these cases came from Group 3. Inhibin α and CAM 5.2 staining did not show this problem. Conclusions We suggest that inhibin α is a useful antibody in diagnosing IUP and HM and in documenting intra-uterine gestations in cases with PIUP because it is a sensitive marker in immunolabelling IT and ST. Combined application of inhibin α and CAM 5.2 might be more useful than βHCG and HPL because the latter showed background staining in one third of the cases.

Placental site nodules or plaques (PSN-Ps) are nodular benign lesions of the intermediate trophoblast (IT) cells in the endometrium, endocervix, superficial myometrium or fallopian tube, occurring after a remote intrauterine pregnancy. We present a study of 25 cases of PSN-Ps These lesions occurred in patients aged 18 to 44 years. Most were discovered incidentally in endometrial curettage specimens. The specimens were received as part of clinical investigations for menorrhagia, per vaginal bleeding or pelvic pain. None of the PSN-Ps was visible grossly. Microscopically, they were mostly multiple, well-circumscribed, oval or plaque-like cellular nodules. The IT cells typically had abundant vacuolated or eosinophilic cytoplasm. The nuclei were irregular, large, hyperchromatic, often degenerate-looking and either mononucleated, multinucleated or multiclefted. Hyalinization surrounding individual or groups of IT cells, or located in the centre of the nodules, was a constant feature in all cases. The lesional cells were strongly immunoreactive to CAM 5.2, 34 beta E12, AE1/AE3, EMA and vimentin. Some cases showed focal positivity to HCG and HPL. PLAP staining was consistently negative. Ultrastructurally, the IT cells showed prominent nuclear variation in size and shape. The abundant, vacuolated cytoplasm contained some rough endoplasmic reticulum and loosely arranged filaments. This study describes the clinicopathological and immunophenotypic features of 25 cases of PSN-Ps including the ultrastructural findings of one case.

Aims To investigate the potential of quantum dot (Qdot) nano-crystals for use as universal immunocytochemical markers that can be visualised by both light and electron microscopy in living and fixed cells. Methods We have used Qdot probes for immunocytochemical detection of somatostatin hormone in fixed human somatostatinoma tumour tissue. Results Qdots applied to etched resin-embedded sections produced highly specific immunolabelling that was able to be detected by widefield fluorescence, super-resolution light microscopy and transmission electron microscopy. The use of adjacent sections allowed correlation between microscopy modalities. The stability of these probes allowed super-resolution light microscopy imaging that correlated well with immunoelectron microscopy. Discussion Streptavidin-conjugated Qdot probes used with a biotinylated secondary antibody labelling protocol produced increased sensitivity and amplification of the labelling signal. The correlation provided by this approach is valuable for assisting with interpretation of antigen localisation.