Ketocyanine-Based Fluorescent Probe Revealing the Polarity Heterogeneity of Lipid Droplets and Enabling Accurate Diagnosis of Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) has gradually become a pronoun for terrifying death owing to its high mortality rate. With the progression of HCC, lipid droplets (LDs) in HCC cells exhibit specific variations such as increased LDs number and decreased polarity, which can serve as the diagnostic target. However, developing an effective method to achieve HCC diagnosis and reveal LDs polarity heterogeneity is still a crucial challenge. Herein, the first high-performance LDs-targeting probe (1) is reported based on ketocyanine strategy with ultrasensitive polarity-responding ability and near-infrared emission. Probe 1 shows excellent sensitivity to polarity parameter Δf (0.027-0.290) with 808-fold fluorescence enhancement and the emission wavelength red-shifts 91 nm. In HCC cells, probe 1 shows a 2.5- to 5.9-fold fluorescence enhancement compared with normal and other cancer cells which exceeds clinical threshold of 2.0, indicating probe 1 can distinguish HCC cells. The LDs polarity heterogeneity is revealed and it displays a sequence, HCC cells < other cancer cells < normal cells, which may provide useful insight to engineer LDs-targeting probes for HCC cell discrimination. Finally, probe 1 realizes accurate HCC diagnosis on the cellular, organ, and in vivo levels, providing a satisfying tool for clinical HCC diagnosis and surgical navigation.

A novel nude mouse model for studying the pathogenesis of endometriosis.

Endometriosis is a common female gynecological disease that is characterized by the presence of functional endometrial tissue outside the uterine cavity. At present, many animal models have been established. However, previous studies consistently use human endometrial tissue implanted in the subcutaneous or abdominal cavity for modeling and rarely use endometrial cells. In the present study, we ascertained whether immortalized stromal and/or epithelial endometrial cells are able to induce subcutaneous endometriosis in nude mice. Mixed human immortalized endometriosis stromal and epithelial cells, but not the cells of Group 1 or Group 2, were successfully constructed and led to endometriotic-like lesions. The endometriosis-like lesions observed in nude mice consisted of endometriosis-like glands lined with columnar epithelial cells and surrounded by stromal cells in the fibrous fatty connective tissue. Immunofluorescence analysis showed that glandular epithelial cells were intensely stained for E-cadherin and cytokeratin 7, and surrounding stromal cells were mildly stained for neprilysin (CD10) and vimentin. Moreover, the cells present in the endometriosis-like lesions were of human origin. Our data indicate that the mixture of human immortalized endometriosis stromal cells and epithelial cells is able to establish subcutaneous endometriosis lesions in nude mice. This model could be used to understand the molecular mechanisms involved in the occurrence and development of endometriosis.

 Primulasurculosa (Primulaceae), a new species from Yunnan, China.

A new species, Primulasurculosa, is described and illustrated. In gross morphology, it is clearly allied to section Petiolares and is most similar to P.taliensis from the group Taliensis, but is distinctive in its indumentum in the throat of the corolla tube, and the markedly stoloniferous habit.

New contributions to Goodyerinae and Dendrobiinae (Orchidaceae) in the flora of China.

Eight new species from China, Cheirostylis chuxiongensis, C. yei, Myrmechis lingulata, M. longii, Bulbophyllum ximaense, B. xizangense, B. retusum and B. pulcherissimum, are described and illustrated. Cheirostylis chuxiongensis differs from C. thailandica by having 5-9 irregular and papillae-like calli on each side in the sac of the lip, epichile with entire lobes, petals narrowly obliquely obovate and an apex that is not recurved. Cheirostylis yei is easily distinguished from its relatives similar by having a long stem, pubescent ovary and sepals, epichile lobes with irregular and undulate margins, a subquadrate callus without teeth in the saccate hypochile. Myrmechis lingulata differs from M. chinensis by having a simple and lanceolate to ligulate lip, glabrous bracts and ovary, oblique and narrowly ovate petals. Myrmechis longii differs from M. pumila by having white-veined leaves, oblong-lanceolate epichile lobes, and viscidium attached to the middle of the caudicle. Bulbophyllum ximaense is easily distinguished from its relatives similar by having distant pseudobulbs, shorter scape, an inflorescence with 9-16 orange-red flowers, shorter lateral sepals with a long acuminate apex, incurved and tubular apical margins, a papillate lip disk and triangular-subulate stelidia. Bulbophyllum xizangense is easily distinguished from its relatives similar by having narrow lanceolate leaves, shorter inflorescence with 1-3 greenish-yellow flowers, falcate-ovoid lateral sepals, a lip with small lateral lobes and 3 keels at the base. Bulbophyllum retusum differs from B. spathulatum by having shorter inflorescence, peduncles with 2 tubular sheaths, dorsal sepals with a retuse apex, lateral sepals with lower edges that are connate to each other and free and divergent toward the apex, obovate petals with an acute or slightly retuse apex. Bulbophyllum pulcherissimum differs from B. lopalanthum by its 5-veined dorsal sepal, ovate-lanceolate lateral sepals, obliquely ovate-oblong petal, erose-toothed margins and obovate lip with a large, oblong basal callus, and an obtuse base. In addition, three species (Bulbophyllum frostii, B. raskotii and B. nematocaulon) are reported for the first time in China.

Three new species of Liparis s.l. (Orchidaceae: Malaxideae) from Southwest China based on morphological characters and phylogenetic evidence.

Liparis aureolabella and L. mengziensis, two new species from the karst region of southwestern China, and L. bingzhongluoensis, a new species from montane region in Yunnan, are described and illustrated. L. aureolabella is easily distinguished from its relatives by having abaxially purple leave with purple reticulate veins prominent adaxially, a lip auriculate at base, and falcate-lanceolate pollinia. Liparis mengziensis is closely related to L. petiolata and L. auriculata, but differs from them by having an ovate to broadly ovate leaf, purple lip and apex connate along the margins. Liparis bingzhongluoensis is similar to Liparis nanlingensis, but the new species is characterized by having a lip with two transparent ridges on its disc, longitudinally concave basal callus and triangular column wings. Phylogenetic analyses based on nuclear ribosomal ITS and plastid matK sequences showed that L. aureolabella and L. mengziensis are nested with L. petiolata or L. auriculata in a monophyletic clade. L. bingzhongluoensis is sister to a clade formed by L. nanlingensis, L. tsii, L. sasakii and L. krameri. Moreover, morphological comparisons strongly support that the three species as separated species newly to science.

Petrocodon wenshanensis, a new species of Gesneriaceae from southwestern China.

A new species of Petrocodon, P. wenshanensis from Yunnan province of southwestern China, is described and illustrated here. P. wenshanensis morphologically closely resembles P. jingxiensis and P. lithophilus, but differs in vegetative and generative characters. Differences between the new species and others Petrocodon species occurring in Yunnan Province are also shown in the identification key.

Four new species of Oreocharis (Gesneriaceae) in Yunnan province, China.

Four new species of Oreocharis (Gesneriaceae) are described and illustrated. These new species grow in pairs in montane forests in Yunnan province, China. One pair grows in Wenshan county, Southeast Yunnan, viz. Oreocharis eriocarpa W.H. Chen & Y.M. Shui and O. wenshanensis W.H. Chen & Y.M. Shui and another pair grows in Yongde county, Southwest Yunnan, viz. O. fulva W.H. Chen & Y.M. Shui and O. lacerata W.H. Chen & Y.M. Shui. Their morphological and geographical relationship with similar species is discussed and the IUCN endangered status is provided, based on the available data.

FoxM1 promotes the migration of ovarian cancer cell through KRT5 and KRT7.

Forkhead box M1(FoxM1) played an important role in the pathogenesis of ovarian cancer, but its downstream molecular network is mysterious. Here, we combined ChIP-seq with RNA-seq analysis and identified 687 FoxM1-binding regions and 182 genes regulated by FoxM1. The above data pointed out that KRT5 and KRT7 were downstream target genes of FoxM1. Next, we used qPCR and Western blot to verify that FoxM1 knockdown inhibited the expression levels of KRT5 and KRT7. We also demonstrated that FoxM1 regulated KRT5 and KRT7 genes expression through binding a consensus AP-2 cis element, and showed that KRT5 and KRT7 deficiency could prevent the migration but not proliferation of SK-OV-3 cells. Finally, tissue microarray results indicated that KRT5 and KRT7 were highly expressed in ovarian cancer and positively correlated with FoxM1 expression. TCGA database showed that high expression of KRT5 and KRT7 could significantly reduce the survival rate of patients with ovarian cancer. The above results clarify the specific downstream molecular network of FoxM1 to promote the pathogenesis of ovarian cancer, and provide a basis experiment for the judgment of ovarian cancer prognosis and the design of drug targets.

Bioinformatic analysis reveals the importance of epithelial-mesenchymal transition in the development of endometriosis.

BACKGROUND: Endometriosis is a frequently occurring disease in women, which seriously affects their quality of life. However, its etiology and pathogenesis are still unclear. METHODS: To identify key genes/pathways involved in the pathogenesis of endometriosis, we recruited 3 raw microarray datasets (GSE11691, GSE7305, and GSE12768) from Gene Expression Omnibus database (GEO), which contain endometriosis tissues and normal endometrial tissues. We then performed in-depth bioinformatic analysis to determine differentially expressed genes (DEGs), followed by gene ontology (GO), Hallmark pathway enrichment and protein-protein interaction (PPI) network analysis. The findings were further validated by immunohistochemistry (IHC) staining in endometrial tissues from endometriosis or control patients. RESULTS: We identified 186 DEGs, of which 118 were up-regulated and 68 were down-regulated. The most enriched DEGs in GO functional analysis were mainly associated with cell adhesion, inflammatory response, and extracellular exosome. We found that epithelial-mesenchymal transition (EMT) ranked first in the Hallmark pathway enrichment. EMT may potentially be induced by inflammatory cytokines such as CXCL12. IHC confirmed the down-regulation of E-cadherin (CDH1) and up-regulation of CXCL12 in endometriosis tissues. CONCLUSIONS: Utilizing bioinformatics and patient samples, we provide evidence of EMT in endometriosis. Elucidating the role of EMT will improve the understanding of the molecular mechanisms involved in the development of endometriosis.

Paraboea wenshanensis, a new species of Gesneriaceae from Yunnan, China.

Paraboea wenshanensis is a new species of Gesneriaceae from Yunnan, China and is described and illustrated here. It is morphologically similar to P. angustifolia, P. martinii and P. glutinosa, but the congeners of this new taxon can be distinguished by several salient characters. A description of P. wenshanensis, together with illustrations and photographs, a distribution map and conservation assessment are presented.

Suppressor of fused (Sufu) promotes epithelial-mesenchymal transition (EMT) in cervical squamous cell carcinoma.

Suppressor of fused is essential for the maximal activation of Sonic Hedgehog signaling in development and tumorigenesis. However, the role of Sufu in cervical carcinoma remains unknown. Here, we report new findings of Sufu in regulating the epithelial-to-mesenchymal transition through the FoxM1 transcriptional modulation by 14-3-3ζ protein in cervical carcinoma. Sufu is overexpressed in cervical squamous cell carcinoma and its level in clinical tumor tissues is positively correlated with 14-3-3ζ. Functionanlly, siSufu remarkably prevents the cancer cell migration and invasion. We further demonstrate that the transcriptional activity of Sufu is increased by FoxM1, of which stability is promoted by 14-3-3ζ. Knockdown FoxM1 decreases the invasion of SiHa cells and reconstitution of Sufu rescues the invasion of these cells.Finally, overexpression of Sufu is significantly associated with differentiation grade, FIGO stage, Depth of stromal invasion and vascular cancer embolus. Our findings highlight a novel role for Sufu in cervical carcinogenesis.

Growth of human colon cancer cells in nude mice is delayed by ketogenic diet with or without omega-3 fatty acids and medium-chain triglycerides.

BACKGROUND: Tumors are largely unable to metabolize ketone bodies for energy due to various deficiencies in one or both of the key mitochondrial enzymes, which may provide a rationale for therapeutic strategies that inhibit tumor growth by administration of a ketogenic diet with average protein but low in carbohydrates and high in fat. MATERIALS AND METHODS: Thirty-six male BALB/C nude mice were injected subcutaneously with tumor cells of the colon cancer cell line HCT116. The animals were then randomly split into three feeding groups and fed either a ketogenic diet rich in omega-3 fatty acids and MCT (MKD group; n=12) or lard only (LKD group; n=12) or a standard diet (SD group; n=12) ad libitum. Experiments were ended upon attainment of the target tumor volume of 600 mm3 to 700 mm3. The three diets were compared for tumor growth and survival time (interval between tumor cell injection and attainment of target tumor volume). RESULTS: The tumor growth in the MKD and LKD groups was significantly delayed compared to that in the SD group. CONCLUSIONS: Application of an unrestricted ketogenic diet delayed tumor growth in a mouse xenograft model. Further studies are needed to address the mechanism of this diet intervention and the impact on other tumor-relevant parameters such as invasion and metastasis.

MAML2 rearrangement in primary pulmonary mucoepidermoid carcinoma and the correlation with FLT1 expression.

INTRODUCTION: Primary pulmonary mucoepidermoid carcinoma (PMEC) is an uncommon neoplasm with remarkable resemblance to mucoepidermoid carcinoma of the salivary glands. The latter has been shown to harbor t(11,19) resulting in MECT1-MAML2 fusion, which may be of diagnostic and prognostic values. However, the importance of such feature in PMEC has not been well studied. METHODS: We detected MAML2 rearrangement using fluorescence in situ hybridization (FISH) in tissue samples from 42 cases of PMEC and 40 of adenosquamous carcinoma (ASC), and the expression of potential downstream targets of MECT1-MAML2, including HES1, FLT1 and NR4A2 with immunohistochemistry (IHC). The findings were then examined regarding the clinicopathological parameters and patient outcomes. RESULTS: FISH analysis revealed MAML2 rearrangement in 50% of the PMEC cases, and such property was prominent in considerable younger patients (33 versus 60 years; p = 0.001) and restricted to cases of low and intermediate grades. IHC analysis showed that FLT1 and HES1 were expressed at lower level in MAML2 rearranged group than MAML2 non-rearranged group (p<0.001 and p = 0.023, respectively). Survival analysis showed significant correlation between MAML2 rearrangement and overall survival (p = 0.023) or disease-free survival (p = 0.027) as well as correlation between FLT1 and overall survival (p = 0.009). CONCLUSIONS: MAML2 rearrangement appears frequent in PMEC and specific with this tumor. Both the presence of MAML2 rearrangement and absence of FLT1 tend to confer a favorable clinical outcome. These findings suggest that molecular detection of MAML2 rearrangement combined with FLT1 may be of important clinical value for PMEC.

The use of mutation-specific antibodies in predicting the effect of EGFR-TKIs in patients with non-small-cell lung cancer.

PURPOSE: We aimed to quantify the epidermal growth factor receptor (EGFR) mutation in tumors and to analyze its prediction of EGFR-tyrosine kinase inhibitor (EGFR-TKI) treatment efficacy in EGFR mutation-positive non-small-cell lung cancer (NSCLC) patients. METHODS: We examined EGFR mutation status in 124 lung cancer samples by direct sequencing and amplification refractory mutation system. Among them, 41 were appropriate to quantify the expression of mutant EGFR proteins using immunohistochemistry (IHC) with mutation-specific antibodies. The quantification was determined by both the staining intensity and the proportion of stained tumor cells. RESULTS: The median progression-free survival (PFS) in patients with a high score for mutant EGFR expression was 18.0 months (95 % CI 16.0-20.0), which was significantly longer than that in patients with a low score (8.0 months; 95 % CI 2.6-13.4; P = 0.048). Such significant association with patients' PFS was also apparent in the proportion of stained tumor cells (median, 19.0 vs. 8.0 months; P = 0.019), but not in the staining intensity (P = 0.787). Among the 41 specimens, 32 were detected EGFR mutation positive by both direct sequencing and ARMS, referring to a relatively high abundance of mutation, and 26 (81.3 %) of them gained a high expression score of mutant proteins as well. Six samples with mutation negative by direct sequencing but positive by ARMS, which showed a low abundance, and 5 (83.3 %) of them also revealed a low expression score. The EGFR mutation quantitative analysis using mutation-specific IHC was moderately consistent with that by molecular-based assays (P = 0.001, kappa value 0.50). CONCLUSIONS: Our results suggest that immunohistochemical analysis with mutation-specific antibodies is a promising approach for quantifying EGFR mutations, and may predict the effect of EGFR-TKI treatment for EGFR mutation-positive NSCLC.

Contrast-enhanced multiphasic CT and MRI findings of adenosquamous carcinoma of the pancreas.

OBJECTIVE: The objective was to retrospectively study computed tomography (CT) and magnetic resonance imaging (MRI) findings of adenosquamous carcinoma of the pancreas (PASC). MATERIALS AND METHODS: Twelve patients (six women and six men; mean age, 61.3 years; range, 47-78 years) who presented with PASC as documented by pathologic examination underwent CT (n=10) or both CT and MRI (n=2) examination. Two radiologists evaluated the images and determined the location, size, margin, internal attenuation or signal intensity, contrast enhancement, and pattern for each tumor. Additionally, the presence of poorly enhanced areas, upstream main pancreatic duct (MPD) dilatation, pancreatic atrophy, and peripancreatic tissue metastasis were evaluated. Images were cross-referenced to surgical and pathologic findings. RESULTS: Masses were distributed throughout the pancreas (head, n=6; body, n=1; and tail, n=5). The tumor size ranged from 2.4 to 5.5 cm with an average size of 3.7 cm. Eight (66.7%) masses were ill defined, and seven (58.3%) were partially exophytic. Twelve (100%) masses showed heterogeneous and poorly enhanced areas. The lesions showed weak (n=5), moderate (n=5), or intense (n=2) progressive enhancement. The diameter of MPD in six patients ranged from 3.0 to 5.0 mm with an average of 3.7 mm. Pancreatic atrophy was not found. In 10 patients (83.3%), masses invaded the peripancreatic tissues. Two patients had metastatic liver disease at presentation. CONCLUSION: PASC typically presented as an ill-defined, hypovascular mass with a poorly enhanced area, exophytic tendency, and peripancreatic tissue invasion. Lack of pancreatic atrophy and mild MPD dilatation were also distinct from common duct pancreatic adenocarcinoma.

Subsequent intra-abdominal fibromatosis mimicking recurrent gastrointestinal stromal tumor.

Intra-abdominal fibromatosis (IAF) commonly develops in patients who had abdominal surgery. In rare instances, it occurs subsequent to gastrointestinal stromal tumor (GIST). This special situation has clinical significance in imatinib era. About 1000 patients with GIST in our institution from 1993 to 2010 were re-evaluated based on their clinical and pathological data, the treatment strategies and the follow-up information. We identified 2 patients who developed IAF after GIST resection. Patient 1 was a 54 year-old male and had 5 cm × 4.5 cm × 3.5 cm jejunal GIST excised on February 22, 1994. Three years later, an abdominal mass with 7 cm × 6 cm × 3 cm was identified. He was diagnosed as recurrent GIST from clinical point of view. After excision, the second tumor was confirmed to be IAF. Patient 2 was a 45-year-old male and had 6 cm × 4 cm × 3 cm duodenal GIST excised on August 19, 2008. One year later, a 4 cm mass was found at the original surgical site. The patient refused to take imatinib until the tumor increased to 8 cm six months later. The tumor continued to increase after 6 months' imatinib therapy, decision of surgical resection was made by multidisciplinary team. The second tumor was confirmed to be IAF with size of 17 cm × 13 cm × 11 cm. Although IAF subsequent to GIST is very rare, it is of clinical significance in imatinib era as an influencing factor for making clinical decision. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1076715989961803.

Multiphasic contrast-enhanced CT and MRI findings of adult mesoblastic nephroma: A report of two cases.

Mesoblastic nephroma (MN) presenting in an adult is extremely rare. The computed tomography (CT) and magnetic resonance imaging (MRI) features of this tumor in adulthood have not been widely reported. We present two additional cases of adult MN and describe the multiphasic contrast-enhanced CT and MRI findings.

[Screening of differentially expressed microRNAs in borderline and malignant gastrointestinal stromal tumors].

OBJECTIVE: Gastrointestinal stromal tumors (GISTs) have a broad spectrum of biological behaviors ranging from benign, borderline and malignant. This study aimed to screen differentially expressed microRNAs (miRNAs) between malignant and borderline GISTs and to investigate the potential role of miRNAs in the malignant transformation of GISTs. METHODS: Six GIST samples including borderline tumors (n = 3) and malignant tumors (n = 3) were collected based on the clinical and pathological characteristics. Total RNA was extracted, followed by miRNA microarray analysis to screen the differentially expressed miRNAs. The most significantly expressed 4 miRNAs were then chosen for further validation by real-time PCR in 22 additional GIST samples. RESULTS: Direct comparison of malignant group versus borderline group revealed 14 significantly and differentially expressed miRNAs (P < 0.05, with a fold change of < 0.5 or > 2). Five miRNAs were up-regulated and nine were down-regulated in the malignant group. Four miRNAs (miR-221, miR-135b, miR-675(*) and miR-218) were most significantly and differentially expressed between the two groups. The differential expression of 2 miRNAs (miR-221 and miR-675(*)) were subsequently confirmed with good concordance by real-time PCR. CONCLUSIONS: The differential miRNA expression profiles between two groups are revealed by miRNA microarray assay, and confirmed by real-time PCR. Among differentially expressed miRNAs, miR-221 and miR-675(*) might be related to the malignant transformation of GISTs, and have a potential value in predicting biological behavior of GISTs.