A Tie2 kinase mutation causing venous malformations increases phosphorylation rates and enhances cooperativity.

The endothelial receptor tyrosine kinase Tie2 plays an important role in vascular formation and maintenance. Mutations in Tie2 lead to vascular malformations, which are painful vascular lesions that cause disfigurement, bleeding, and thrombosis. R849W Tie2 is the most common mutation implicated in an inherited form of vascular malformations and has been shown to be activating, though little is known about the kinetic features of catalysis. Here we undertake a steady-state kinetic analysis of heterologously expressed and purified wild type (WT) and R849W Tie2. While the catalytic efficiencies of the two forms are not significantly different, the observed maximal rate of phosphorylation, kcat,obs, is > 3-fold higher for R849W Tie2 compared to WT. Notably, steady-state catalysis by R849W Tie2 has more striking sigmoidal features compared to WT, suggesting enhanced positive cooperativity. We propose that activating catalytic features are one important consequence of the R849W mutation, though likely other factors such as increased protein binding affinity also contribute to the phenotypes observed in patients.

Induced sputum deposition improves diagnostic yields of pulmonary alveolar proteinosis: A clinicopathological and methodological study of 17 cases.

Pulmonary alveolar proteinosis (PAP) is a rare diffuse lung disease characterized by the accumulation of intra-alveolar lipoprotein-like surfactants. Lung core biopsy and bronchoalveolar lavage (BAL) fluid are currently the two major sources of sampling for diagnosis. In the present study, we assessed the value of induced sputum in diagnosing PAP by transmission electron microscopy and examined the PAP 2-year death rate in Asians. Transmission electron microscopy was performed on the samples from 17 patients with PAP, 13 patients with inflammatory lung diseases, and 13 healthy adults. The PAP patients were followed up for 3-156 months, and inflammatory lung diseases patients or healthy adults for 12-36 months. The ultrastructural features including diagnostic lamellar bodies of induced sputum deposition (ISD) samples were similar to that of the BAL fluid sediment. However, the rates of lamellar bodies were 73.7% in the ISD group, significantly higher than the spontaneous sputum deposition (SSD) group (42.1%, P < .0487) and similar to the BAL sediment (76.2%) and the lung biopsy (54.5%) groups. The overall 2-year death rate of our PAP patients was 17.6% (3/17), not statistically different from the healthy adults and patients with inflammatory diseases (0/13, P = .237 for both). ISD may be the preferred non-invasive sampling method for diagnosing PAP by electronic microscopy because of the higher diagnostic yield than SSD. The diagnostic yields of this noninvasive method were similar to that of lung core biopsy and BAL.

ß-Catenin overexpression is associated with gefitinib resistance in non-small cell lung cancer cells.

BACKGROUND: Acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) presents great challenges in the treatment of non-small cell lung cancer (NSCLC) patients, while the mechanisms are still not well understood. The ß-catenin signaling pathway has been found to be associated with chemoresistance and can activate the EGFR and its downstream pathways. This study aimed to investigate the role of ß-catenin in acquired resistance to EGFR-TKIs in NSCLC cell lines. METHODS: The expression and transcriptional activity of ß-catenin were measured in both the NSCLC cell line PC9 and its sub-line PC9/AB(2) which has acquired resistance to gefitinib. Knockdown and overexpression of ß-catenin in the PC9/AB(2) and PC9 cells were performed. The cell survival rate and the activation of the EGFR and its downstream pathways were detected in the two cell lines after transfection. RESULTS: Nuclear translocation of ß-catenin was increased in the PC9/AB(2) cells and the baseline expression of members of the ß-catenin signaling pathway was also higher in the PC9/AB(2) cells. Knocking down the expression of ß-catenin increased the sensitivity of the PC9/AB(2) cells to gefitinib by blocking the activation of the EGFR downstream pathways, while ß-catenin overexpression improved PC9 cells resistance to gefitinib by enhancing the activation of the EGFR and its downstream signaling. CONCLUSION: ß-catenin plays an important role in acquired resistance to EGFR-TKIs in NSCLC cell lines and may be a potential therapeutic target for NSCLC patients who have failed to respond to targeted therapy.

[Clinical and histopathological features of colorectal sessile serrated adenoma/polyp and its differential diagnosis].

OBJECTIVE: To investigate clinicopathological characteristics of colorectal sessile serrated adenoma/polyp (SSA/P) and its differential diagnosis from other serrated lesions. METHODS: Clinicopathological features of all cases of colorectal serrated lesions from 5 209 colorectal biopsy samples at Shanghai Tongji Hospital from 2008 to 2013 were reviewed. Three hundred and fifty-three cases of serrated lesions were erolled in the study. Morphological features of SSA/P were investigated with an emphasis on histologic criteria for diagnosis and a literature review was performed. RESULTS: Three hundred and fifty-three cases of serrated lesions were identified, including 25 SSA/P (7.1%), 278 hyperplastic polyp (HP, 78.8%), and 44 traditional serrated adenoma (TSA, 12.5%). Twenty-five patients with SSA/P consisted of 16 males and 9 females with a mean age of 62.2 years (aged 34-84 years) and the lesions involved sigmoid colon (14 cases), ascending colon (9 cases), rectum (1 case) and transverse colon (1 case). Grossly, the majority of SSA/P was sessile with an averaged size of 0.73 cm. Histologically, typical SSA/P had elongated crypts with prominent serration and distorted crypts architecture. The detection rates of crypts dilatation and branching in SSA/P and HP were 100% (25/25) and 24% (12/50, P < 0.01), 72% (18/25) and 4% (2/50, P < 0.01), respectively. Morphological features observed only in SSA/P included L-shaped crypts (48%, 12/25), pseudo infiltration of mucosa muscle (16%, 4/25), atypical nuclei (32%, 8/25), and increased mucus secretion (24%, 6/25). CONCLUSIONS: SSA/P microscopically shows prominent serration and abnormal architectures of crypts. Complete tissue sectioning and correct embedding are helpful for the diagnosis. SSA/P without cytological dysplasia should be distinguished from HP, especially those with only a few distorted crypts.

Susceptibility of Cd availability in microplastics contaminated paddy soil: Influence of ferric minerals and sulfate reduction.

The combined contamination of cadmium (Cd) and microplastics (MPs) in paddy soil always occurred, while its influence on Cd availability remained unclear. This study investigated the Cd availability in Cd-MPs co-contaminated paddy soil in consideration of both ferric minerals and sulfate reduction under flooding conditions. The presence of MPs resulted in a higher Cd releasing risk, as represented by the increase in the available Cd and decrease in Fe-Mn oxide-bound Cd contents, especially on the 7th and 14th days based on the sequential extraction results. MPs facilitated the formation of Fe-organic ligands, which accelerated the reductive dissolution of iron minerals but decreased the amounts of amorphous iron minerals due to the release of dissolved organic substances into pore water. Furthermore, MPs promoted the relative abundance of sulfate-reducing bacteria (such as Streptomyces and Desulfovibrio genera), thus increasing the contents of reductive S species, which was advantageous to the co-precipitation of Fe, S, and Cd on the surface of MPs based on our experimental and statistical results. Taken together, both iron and sulfate reduction under anaerobic conditions played a critical role in Cd mobilization in Cd-MPs co-contaminated paddy fields.

Comprehensive distribution characteristics and factors affecting the migration of chromium in a typical chromium slag-contaminated site with a long history in China.

The contamination of abandoned chromium slag-contaminated sites poses serious threats to human health and the environment. Therefore, improving the understanding of their distribution characteristics and health risks by multiple information is necessary. This study explored the distribution, accumulation characteristic, and the role in the migration process of chromium. The results showed that the contents of total Cr and Cr (VI) ranged from 12.00 to 7400.00 mg/kg, and 0.25 to 2160.00 mg/kg, respectively. The average contents of both total Cr and Cr (VI) reached the highest value at the depth of 7-9 m, where the silt layer retaining total Cr and Cr (VI) was. The spatial distribution analysis revealed that the total contamination area percentages of total Cr and Cr (VI) reached 7.87% and 90.02% in the mixed fill layer, and reduced to 1.21% and 34.53% in the silty layer, and the same heavily polluted areas were located in the open chromium residue storage. Soil pH and moisture content were the major factors controlling the migration of total Cr and Cr(VI) in soils. Results of probabilistic health risk assessment revealed that carcinogenic risk was negligible for adults and children, and the sensitive analysis implied that the content of Cr(VI) was the predominant contributor to carcinogenic risk. The combination of chemical reduction and microbial remediation could be the feasible remediation strategy for soil Cr(VI) pollution. Overall, this study provides scientific information into the chromium post-remediation and pollution management for various similar chromium-contaminated sites.

[Effect of Wnt signaling suppression on gefitinib in non small cell lung cancer cell lines].

OBJECTIVE: To explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib, and its related mechanisms. METHODS: PC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib, and the proliferation index was measured using CCK8 kit. The members of Wnt signaling pathway were detected by Western blot. Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of ß-catenin. ß-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription, followed by treatment with different concentrations of gefitinib. Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of ß-catenin. RESULTS: Treating with different concentrations of gefitinib, the resistance of PC9/AB2 cells to gefitinib was significantly increased (P < 0.05). The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t = 24.590, P = 0.000). TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t = 4.983, P = 0.008). Compared with the negative control group, apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P < 0.05). The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression, although other proteins showed no significant alterations. CONCLUSION: Suppressing the activity of Wnt signaling can partly reverse the celluar resistance to gefitinib in non small cell lung cancer.

Improved multivariate modeling for soil organic matter content estimation using hyperspectral indexes and characteristic bands.

Soil organic matter (SOM) is a key index of soil fertility. Calculating spectral index and screening characteristic band reduce redundancy information of hyperspectral data, and improve the accuracy of SOM prediction. This study aimed to compare the improvement of model accuracy by spectral index and characteristic band. This study collected 178 samples of topsoil (0-20 cm) in the central plain of Jiangsu, East China. Firstly, visible and near-infrared (VNIR, 350-2500 nm) reflectance spectra were measured using ASD FieldSpec 4 Std-Res spectral radiometer in the laboratory, and inverse-log reflectance (LR), continuum removal (CR), first-order derivative reflectance (FDR) were applied to transform the original reflectance (R). Secondly, optimal spectral indexes (including deviation of arch, difference index, ratio index, and normalized difference index) were calculated from each type of VNIR spectra. Characteristic bands were selected from each type of spectra by the competitive adaptive reweighted sampling (CARS) algorithm, respectively. Thirdly, SOM prediction models were established based on random forest (RF), support vector regression (SVR), deep neural networks (DNN) and partial least squares regression (PLSR) methods using optimal spectral indexes, denoted here as SI-based models. Meanwhile, SOM prediction models were established using characteristic wavelengths, denoted here as CARS-based models. Finally, this research compared and assessed accuracy of SI-based models and CARS-based models, and selected optimal model. Results showed: (1) The correlation between optimal spectral indexes and SOM was enhanced, with absolute value of correlation coefficient between 0.66 and 0.83. The SI-based models predicted SOM content accurately, with the coefficient of determination (R2) and root mean square error (RMSE) values ranging from 0.80 to 0.87, 2.40 g/kg to 2.88 g/kg in validation sets, and relative percent deviation (RPD) value between 2.14 and 2.52. (2) The accuracy of CARS-based models differed with models and spectral transformations. For all spectral transformations, PLSR and SVR combined with CARS displayed the best prediction (R2 and RMSE values ranged from 0.87 to 0.92, 1.91 g/kg to 2.56 g/kg in validation sets, and RPD value ranged from 2.41 to 3.23). For FDR and CR spectra, DNN and RF models achieved more accuracy (R2 and RMSE values ranged from 0.69 to 0.91, 1.90 g/kg to 3.57 g/kg in validation sets, and RPD value ranged from 1.73 to 3.25) than LR and R spectra (R2 and RMSE values from 0.20 to 0.35, 5.08 g/kg to 6.44 g/kg in validation sets, and RPD value ranged from 0.96 to 1.21). (3) Overall, the accuracy of SI-based models was slightly lower than that of CARS-based models. But spectral index had a good adaptability to the models, and each SI-based model displayed the similar accuracy. For different spectra, the accuracy of CARS-based model differed from modeling methods. (4) The optimal CARS-based model was model CARS-CR-SVR (R2 and RMSE: 0.92 and 1.91 g/kg in validation set, RPD: 3.23). The optimal SI-based model was model SI3-SVR (R2 and RMSE: 0.87 and 2.40 g/kg in validation set, RPD: 2.57) and model SI-SVR (R2 and RMSE: 0.84 and 2.63 g/kg in validation set, RPD: 2.35).

Soil organic carbon content increase in the east and south of China is accompanied by soil acidification.

Increased soil organic carbon (OC) in China has been reported in the past two decades, suggesting the sequestration of atmospheric carbon dioxide into soil, mitigating climate change and improving soil health. On the other hand, soil pH decrease had also been reported nationwide. If the two are related, the strategy of increasing soil OC could negatively affect soil quality for food production and the environment. We investigate this thread based on large-scale soil survey data from two provinces with typical soil and cropping patterns in the east and south of China, Jiangsu (102,600 km2) and Guangdong (177,900 km2). The data include >5000 observations from soil surveys conducted over the past four decades, i.e., the 1980s, 2006-2007, and 2010-2011. Using spatiotemporal modelling, we show that across Jiangsu province, the topsoil OC on average has increased from 8.5 g kg-1 to 9.9 g kg-1 from 1980 to 2000 and a further increase to 12.6 g kg-1 in 2010. This increase was accompanied by a decrease in average pH from 7.63 to 6.90. In Guangdong, there was an overall increase in average topsoil OC content from 14.2 g kg-1, 16.5 g kg-1, and 20.2 g kg-1 with a decrease in average pH from 5.58, 4.90, and 4.98. Based on the spatiotemporal modelling results, the structural equation modelling analysis shows that OC and pH changes were significantly correlated and linked by increased soil N content. On croplands, soil N content was mainly attributed to N fertiliser application. The pH decrease was particularly significant in the east of China where the soils were neutral in pH. We recommend that more revolutionary means be taken to sequestrate atmospheric carbon into soil as the current OC increase due to increasing crop productivity via a high rate of nitrogen application may have a potential acidification effect.

Primary Clear Cell Sarcoma of the Ileum: A Case Report With Next-Generation Sequencing Analysis.

As the concept of clear cell sarcoma-like tumor or malignant gastrointestinal neuroectodermal tumor (CCS-LT/MGNET) has been widely accepted, primary CCS of the gastrointestinal tract (CCS-GI) is becoming a rare entity. In this article, we describe a case of primary CCS-GI that occurred in the ileum of a 65-year-old male to further illustrate its rare occurrence. Similar to CCS of soft tissue (CCS-ST), the tumor was composed of spindled to epithelioid cells displaying fascicular, nested, or pseudopapillary arrangement. The tumor cells had large round to ovoid nuclei with vesicular chromatin and prominent nucleoli, containing eosinophilic to pale cytoplasm. In contrast to CCS-LT/MGNET, immunohistochemical study also showed variable positivity of HMB45, melan A, and MiTF besides the strong and diffuse staining of S100 protein and SOX10. Fluorescence in situ hybridization (FISH) using fusion probes identified EWSR1 and ATF1 genes rearrangement. Next-generation sequencing (NGS) analysis further revealed EWSR1 exons9/8-ATF1 exon4 and ATF1 exon3- EWSR1 exon11 fusion genes. CCS-GI and CCS-LT/MGNET possibly represent 2 related entities of the same spectrum, which differentiate along 2 different pathways.

Ion Binding Properties of a Naturally Occurring Metalloantibody.

LT1009 is a humanized version of murine LT1002 IgG1 that employs two bridging Ca2+ ions to bind its antigen, the biologically active lipid sphingosine-1-phosphate (S1P). We crystallized and determined the X-ray crystal structure of the LT1009 Fab fragment in 10 mM CaCl2 and found that it binds two Ca2+ in a manner similar to its antigen-bound state. Flame atomic absorption spectroscopy (FAAS) confirmed that murine LT1002 also binds Ca2+ in solution and inductively-coupled plasma-mass spectrometry (ICP-MS) revealed that, although Ca2+ is preferred, LT1002 can bind Mg2+ and, to much lesser extent, Ba2+. Isothermal titration calorimetry (ITC) indicated that LT1002 binds two Ca2+ ions endothermically with a measured dissociation constant (KD) of 171 µM. Protein and genome sequence analyses suggested that LT1002 is representative of a small class of confirmed and potential metalloantibodies and that Ca2+ binding is likely encoded for in germline variable chain genes. To test this hypothesis, we engineered, expressed, and purified a Fab fragment consisting of naïve murine germline-encoded light and heavy chain genes from which LT1002 is derived and observed that it binds Ca2+ in solution. We propose that LT1002 is representative of a class of naturally occurring metalloantibodies that are evolutionarily conserved across diverse mammalian genomes.

GPRC5A reduction contributes to pollutant benzo[a]pyrene injury via aggravating murine fibrosis, leading to poor prognosis of IIP patients.

Air pollution exposure is recently reported to be one of the drivers of exacerbation in idiopathic pulmonary fibrosis (IPF). But there was a lack of direct evidence between pollution and lung fibrosis. Here, our data show effects of pollutant benzo[a]pyrene (BaP) and protein G-protein-coupled receptor family C group 5 type A (GPRC5A) on pulmonary fibrosis, which might help limit potential pollutant injury and disease progression. We cross-referenced epithelial differentially-expressed-genes (DEGs) from pollutant injury and published experimental fibrosis and IPF patients' data, top common-DEG (CO-DEG) GPRC5A was identified as a potential link between exposure-damage and fibrogenesis. The role of GPRC5A was evaluated under BaP exposure, in idiopathic interstitial pneumonia (IIP) tissue-array and via CRISPR/Cas9 knockout mice (Gprc5a-/-). BaP exposure enhanced bleomycin (BLM)-induced murine pulmonary fibrosis with increased Fibronectin and α-SMA expression in primary fibroblasts, thickened respiratory membrane and damaged alveolar type II cell, combined with Gprc5a decline in fibrotic mass. GPRC5A mRNA reduced after 10-14 days' BaP exposure in human epithelial cell A549. GPRC5A protein was further found to decrease in IIP epithelium, especially hyperplastic regions. A high epithelial GPRC5A expression score was positively associated with long survival time (R = 0.34) while negatively with high age (R = -0.4) and IIP type IPF (R = -0.5). Low GPRC5A expression predicts poor prognosis (HR = 4.5). Gprc5a depletion aggravated mortality rate (50%) with increased collagen deposition and myofibroblast activation under BLM treatment and exacerbated BaP injury in lung remodeling. Vitamin metabolic imbalance and Mitofusion2 (Mfn2) or Opa1-regulated mitochondrial dynamics were deduced to contribute to Gprc5a depletion and fibrogenesis. Pollutant BaP exposure worsens murine fibrosis and myofibroblast activation via GPRC5A reduction in the damaged epithelium. GPRC5A deficiency was first confirmed to contribute to both poor prognosis of IIP patients and fibrogenesis in murine model; thus, GPRC5A could serve as a novel therapeutic target in pollutant injury and pulmonary fibrosis.

Performance of median kriging with robust estimators of the variogram in outlier identification and spatial prediction for soil pollution at a field scale.

Median kriging with robust estimators of the variogram has been proposed in literature to reduce the influences of outliers in spatial data of soil pollution, because median kriging can utilize outliers in spatial prediction and robust estimators can overcome the bias caused by outliers. However, performance of the method at a field scale remains unknown. This study compared the method in two case studies of soil Pb pollution with two other commonly used methods for outlier identification, including box-plot and standardized kriging prediction error (SKPE), and with two classical geostatistical approaches for spatial prediction, including kriging with and without outliers. One case was based on data with 359 samples collected in an area of 14.5km2 in Jura, Swiss. The other was based on data with 242 samples collected in an area of 2.8km2 in Zhuzhou, China. Results showed that the method identified both global and local outliers, while the method did not identify all global outliers based on the box-plot. For the Jura data which were more seriously affected by outliers than the Zhuzhou data, the method identified 49 outliers, sharing 39 with SKPE which identified a total of 46 outliers. For the Zhuzhou data, the method found just three outliers, much fewer than the 12 outliers identified by SKPE. In the case of Jura, kriging prediction with outliers winsorized by the method was negligibly more accurate than prediction without outliers identified by SKPE, e.g., 0.15% in terms of root mean square error (RMSE). However, in the case of Zhuzhou, the former prediction was slightly less accurate than the latter, e.g., 2.39% in terms of RMSE. This study suggested that the method performed well for data which were seriously affected by outliers, but not so well for data slightly affected by outliers.

[Principal Component Analysis and Ecological Risk Assessment of Heavy Metals in Farmland Soils around a Pb-Zn Mine in Southwestern China].

The farmland soil around a Pb-Zn mine in southwestern China was studied. One hundred forty-nine surface soil samples were taken from 0-20 cm depth, and the contents of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn were tested. The correlations among these heavy metals were studied with multivariate analysis, and the potential sources of the metals were identified. The environmental risk of the metals was evaluated with the Potential Ecological Risk Index method. The results showed that the amounts of Cd, Pb, and Zn were at relatively high level, with average concentrations of 15.56, 419.4, and 933.4mg·kg-1 respectively, indicating the soil was heavily polluted. The average concentrations of Hg and As were 0.13 and 37.3mg·kg-1, suggesting moderate soil pollution. The average concentrations of Cu, Ni, and Cr were lower than Yunnan soil background values. The multivariate analysis suggested that the sources of Cd, Pb, Zn, Hg, and As were similar and came mainly from smelting activities in the mining area. The sources of Cu, Ni, and Cr were similar and can be attributed to natural sources. The comprehensive potential ecological risk index was 2294.8, which suggested a high potential ecological risk. In general, the farmland soils in the research area were polluted seriously by the mining and industrial activities.

The expression and clinical relevance of PD-1, PD-L1, and TP63 in patients with diffuse large B-cell lymphoma.

Latest study showed that a novel translocation between programmed cell death ligand 1 (PD-L1) (cluster of differentiation 274) and TP63 (tumor protein 63) can be found in diffuse large B-cell lymphoma (DLBCL), resulting in their conjunct overexpression in tumor cells at RNA level. However, the expressed pattern of these 2 genes at protein level in DLBCL remains largely unknown, and the clinical relevance of PD-L1 and TP63 expression in DLBCL are also unclear.Tumor tissues from 76 Chinese DLBCL patients were immunostained for programmed cell death 1 (PD-1), PD-L1, and TP63 using the EnVision system. Clinical relevance of PD-1, PD-L1, and TP63 in 74 DLBCL were analyzed by chi-square test, the Kaplan-Meier curves with log rank test, and Cox's proportional hazards regression model.PD-1 was mainly expressed in tumor-infiltrating lymphocytes (TILs) of 39.5% patients. PD-L1 was expressed in tumor cells of 26.3% patients, and TP63 was immunostained in nucleoli of tumor cells of 31.6% cases. PD-1 expression was significantly associated with the patients' gender and B symptoms (P = 0.032, P = 0.026). DLBCL with PD-L1 or TP63 expression in tumor cells showed low International Prognostic Index (IPI) score (P = 0.007, P = 0.009). PD-1 TILs was related to prolonged overall survival rate (OS) of DLBCL patients (P = 0.02), whereas PD-L1 expression was associated with worse clinical outcome of patients (P = 0.049). Immunoreactivity of TP63 was not correlated with patients' survival time. Besides, PD-1 expression, patients' age, Ann Arbor stage, and IPI score were significant prognostic markers for OS, but PD-L1 and TP63 had no prognostic significance.PD-1, PD-L1, and TP63 are frequently expressed in DLBCL. PD-1/PD-L1/TP63 blockade may be a potential therapeutic strategy for some patients.

Enhanced colonic tumorigenesis in alkaline sphingomyelinase (NPP7) knockout mice.

Intestinal alkaline sphingomyelinase (alk-SMase) generates ceramide and inactivates platelet-activating factor (PAF) and was previously suggested to have anticancer properties. The direct evidence is still lacking. We studied colonic tumorigenesis in alk-SMase knockout (KO) mice. Formation of aberrant crypt foci (ACF) was examined after azoxymethane (AOM) injection. Tumor was induced by AOM alone, a conventional AOM/dextran sulfate sodium (DSS) treatment, and an enhanced AOM/DSS method. ß-Catenin was determined by immunohistochemistry, PAF levels by ELISA, and sphingomyelin metabolites by mass spectrometry. Without treatment, spontaneous tumorigenesis was not identified but the intestinal mucosa appeared thicker in KO than in wild-type (WT) littermates. AOM alone induced more ACF in KO mice but no tumors 28 weeks after injection. However, combination of AOM/DSS treatments induced colonic tumors and the incidence was significantly higher in KO than in WT mice. By the enhanced AOM/DSS method, tumor number per mouse increased 4.5 times and tumor size 1.8 times in KO compared with WT mice. Although all tumors were adenomas in WT mice, 32% were adenocarcinomas in KO mice. Compared with WT mice, cytosol expression of ß-catenin was significantly increased and nuclear translocation in tumors was more pronounced in KO mice. Lipid analysis showed decreased ceramide in small intestine and increased sphingosine-1-phosphate (S1P) in both small intestine and colon in nontreated KO mice. PAF levels in feces were significantly higher in the KO mice after AOM/DSS treatment. In conclusion, lack of alk-SMase markedly increases AOM/DSS-induced colonic tumorigenesis associated with decreased ceramide and increased S1P and PAF levels.

Histology subtypes and polyp size are associated with synchronous colorectal carcinoma of colorectal serrated polyps: a study of 499 serrated polyps.

Sessile serrated adenomas (SSA) and traditional serrated adenomas (TSA) are considered as precursors of colorectal cancer, and are often diagnostic challenges. Their true prevalence is masked by significant inter-observer variations. To investigate the true prevalence and synchronous colorectal carcinoma (sCRC) of colorectal serrated polyps (CSP) and their associated factors, we first retrospectively identified all colorectal polyps collected at our institution between June 1995 and May 2013. After centrally reclassifying all CSP to reduce inter-observer variations, Chi-square tests and logistic regression analyses were used to analyze the potential factors. Among the included 5501 colorectal polyps, 499 CSP of 428 patients were identified and studied, including 353 hyperplastic polyps (HP, 70.7%), 80 SSA (16.0%), 61 TSA (12.2%) and 5 mixed polyp (1.0%). Diagnostic disagreements were found in 68 CSP (13.63% of CSP). SSA and TSA were more often larger than 5 mm and in proximal colon than HP. SSA were also more likely associated with older age (p=0.005), size ≥5 mm (p<0.001) and ≥3 polyps (p=0.004) than HP in distal colon, but only more likely associated with older age (p=0.006) in proximal colon. Multivariate regression analysis demonstrated that CSP with sCRC, compared with CSP without sCRC, were linked to CSP size ≥1 cm (vs <1 cm, odds ratio [OR] 4.412, 95% confidence interval [CI] 1.684-11.556, P=0.003) and a diagnosis of SSA or TSA (vs HP, OR 6.194, 95% CI 1.870-20.513, P=0.003 and OR 6.754, 95% CI 1.981-23.028, P=0.002, respectively), but not age, gender, polyp number and polyp shape. SSA and TSA are similarly often associated with sCRC (P=0.460). In conclusion, histology subtypes and polyp size may serve as markers for sCRC of CSP. SSA and TSA may warrant careful endoscopic examinations and similar follow-up intervals.

Pulmonary vascular malformation complicating cryptococcal pneumonia in an immunocompetent patient.

An immunocompetent 50-year-old male presented with slight cough and occasional lung congestion. The radiologic findings included diffuse, bilateral reticular and one nodular opacity at the upper lobe of right lung without clear margin. A wedge resection of the lesion showed disordered distribution of the medium-sized vessels and arterioles, several arterioles densely gathered including a few occlusive arterioles, or medium veins dilated with irregular and elongated cavity, indicating the existence of vascular malformation. Interestingly, near to the malformed vessels, a large area of necrosis with granulomatous inflammation was found. Of note, numerous intracytoplasmic organisms with a nucleus, a wall and a thick capsule, were free in the alveoli or located within the macrophages and polykaryocytes, suggesting cryptococci infection. This is to our best knowledge the first case showing concurrent vascular malformation and local pulmonary cryptococcosis, and vascular malformation was likely an important pathological predisposing factor for local pulmonary cryptococcosis infection.