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The accuracy of the segmentation of the target lesion and at-risk surrounding organs is important for cervical cancer patients treated with three-dimensional (3D) brachytherapy. However, the nature of brachytherapy, organ deformities, metal induced artifacts caused by applicators, and limited operating time make this process a challenge. Deep learning segmentation has recently emerged as an approach to this problem. The basic concept proposed in this paper is accurate segmentation of a pelvic computed tomography image dataset such that patients can obtain more accurate radiotherapy. In this work, we propose a solution based on 3D U-Net and Long Short-Term Memory (LSTM). The model was trained and tested on a computed tomography pelvic image dataset comprising 51 patients who underwent 3D brachytherapy. The organs that required segmentation included the bladder, bowels, sigmoid colon, rectum, and uterus. We also used self-ensemble to improve segmentation accuracy. The Dice coefficient was used as the evaluation metric to determine the segmentation results for all of the organs under consideration. The proposed model was evaluated using organ segmentation obtained from 10 patients with newly diagnosed stage I-IVA cervical cancer. The test results showed that segmentation conducted using the proposed model resulted in the following Dice coefficient values (mean, ± standard deviation): 87 (± 6.3) %, 72 (± 9.1) %, 72 (± 7.8) %, 95 (± 3.5) %, 86 (± 4.9) %, 77 (± 8.4) %, 73 (± 10.2) %, and 93 (± 3.5) % for the HRCTV, GTV, bowels, foley, bladder, rectum, sigmoid colon, and uterus, respectively. This method shows that the combination of 3D U-Net and LSTM and self-ensemble post-processing has high potential for segmentation of a pelvic computed tomography dataset.

In brachytherapy, the segmentation accuracy of the target tumor and surrounding organs is very important. In recent years, deep learning models have improved the performance of organ segmentation and have been widely used. However, it is still a huge challenge for some organs with variable shapes. The basic idea of the work presented in this paper is to accurately divide the rectal computed tomography image dataset so that patients can obtain more accurate brachytherapy. In this work, we used 3D U-Net and Long ShortTerm Memory (LSTM) to improve the accuracy of rectal segmentation. This model was trained and tested on the rectal computed tomography image dataset, which contains 51 patients undergoing radiation therapy. The dice coefficient is used as the evaluation index in all results of organ segmentation. After experiments are done, it can be seen that the proposed method has good performance in rectal segmentation.

Intravenous infusion or intra-articular administration of vancomycin is widely used to treat septic arthritis. However, the cytotoxic effects of vancomycin at cellular and molecular levels have yet to be fully elucidated in chondrocytes. Primary human chondrocytes were cultured in media supplemented with vancomycin at a series of concentrations (0, 100, 500, 1000, and 5000 μg/mL) to study the dose-dependent toxicity. Since inflammatory chondrocytes are more catabolic than normal cells, the influence of vancomycin (500 μg/mL) on interleukin-1beta (IL-1β)-stimulated chondrocytes was evaluated. Exposure of chondrocytes to a high dose of vancomycin (≥1000 μg/mL) resulted in chondrotoxicity. Non-lethal dose of vancomycin (500 μg/mL) did not change the viability, morphology or glycosaminoglycan content of the normal or IL-1β-stimulated chondrocytes, or affect the mRNA levels of extracellular matrixes (aggrecan, collagen type I, II, and X) and pro-inflammatory cytokines (IL-1β, TNF-α, IL-6, IL-10, MMP-3, MMP-9, and TIMP-1). ELISA revealed that vancomycin exposure did not affect the levels of PGE2, IL-1β, MMP-3, or MMP-9 in normal and IL-1β-stimulated cells. Western blotting further showed that vancomycin did not affect IκB and p50 to initiate the NF-κB pathway or COX-2 synthesis. Vancomycin at a low dose (500 μg/mL) did not have a deleterious effect on chondrocytes.

Background/purpose: We assessed the mobility of single-root teeth by using Miller's mobility index (MMI) and to analyze the validity of MMI for the diagnosis of periodontitis. Materials and methods: A total of 30 patients were included and the Spearman correlation coefficient was used to assess the correlation between MMI, clinical attachment level (CAL), and probing depth (PD). The validity of MMI for the diagnosis of the severity of periodontitis was evaluated using the receiver operating characteristic (ROC) curve, area under curve (AUC) value, positive predictive value (PPV). Results: Strong correlations were observed between MMI and CAL (r = 0.92) and between MMI and PD (r = 0.76). When the CAL = 3–4 mm and CAL ≥5 mm groups were pooled together, the AUC value was 0.81. The AUC was 0.86 for diagnosis with MMI in the CAL ≥5 mm group. A PPV of 100% was achieved for all grades when MMI >1. When the teeth with PD ≥ 5 to

OBJECTIVES NF-κB plays a crucial role in collagen overproduction in dihydropyridine-induced gingival overgrowth (DIGO) fibroblasts. We aim to investigate the role of the kappa B (IκB) kinase (IKK)-NF-κB pathway and downstream collagen type I (Col I) synthesis in DIGO cells and to demonstrate the therapeutic strategy of interference of this pathway with proteasome inhibitors. METHODS Gingival fibroblasts from DIGO (n = 5) and healthy (n = 5) patients were selected and stimulated with IL-1β, nifedipine, or both. All experiments were run in triplicate and independently for each primary cell sample. RESULTS The results demonstrated that both drugs additively mediated NF-κB activity by activating IKKα/β phosphorylation. They also triggered nuclear translocation of NF-κB, Rela, and p50 (*p

Industry 4.0 is gaining more attention from the public, and thus the correlation between factories and nearby environmental pollution sources is a subject worth in-depth research. Among environmental issues, Particulate Matter2.5 (PM2.5) has received considerable attention in recent years from academic units and governments, and one of the secondary PM2.5 sources is the complex chemical reaction of exhaust gases emitted from factories and ammonia (NH3), with NH3 mostly coming from stock farming. Therefore, the correlation between stock farming data and pollutionsources emitted from factories can be examined by using an artificial neural network (ANN). The first target of this study is to investigate the correlation of factory air pollution source data and stock farming data nearby air monitoring stations to the annual mean PM2.5 concentration of nearby air monitoring stations. Second, the study uses Tensorflow to build an ANN model to analyze whether the industrial and stock farming data have an effect on the PM2.5 concentration. Weather data are taken in this experiment to learn about the correlation. The experimental results show that the Spearman’s correlation coefficient of the factory emitted air pollution data and stock farming data nearby air monitoring stations for the annual mean PM2.5 concentration is 0.6 to 0.9, representing positive correlation. The ANN experiment shows the annual mean PM2.5 concentration classification model with industrial data plus stock farming data plus weather data, in which the ANN classification accuracy is 0.75 as validated by mean square error (MSE) methods. Compared with the ANN classification model only with weather data, the MSE classification accuracy is 1.5. According to the two experiments, the industrial factor and stock farming factor are items that may influence the PM2.5 concentration change.

There are many influential factors in PM2.5, reducing the emission of PM2.5 is one of international subjects. In recent years, it is indicated that one of the sources of secondary PM2.5 is the complex chemical reaction between NH3 and air pollutants (VOCs, particulate matter, NOx, SOx). The Committee on Agriculture of FAO indicates that 64% of NH3 emission on the earth surface is derived from stock raising which motivates this study to discuss following two subjects based on Open Government Data. Subject 1 calculates the effect of the controlled air pollutants (VOCs, particulate matter, NOx, SOx) and the quantity of livestock (e.g. pigs, chickens and so on) nearby the air monitoring stations on the annual mean of PM2.5. Subject 2 uses Apache Spark as Cloud computing platform, the air monitoring stations are geographically clustered by K-medoids to calculate the Spearman's correlation coefficient of pollution source and PM2.5 of each cluster. The experimental results show that the monitoring station with more air pollutants and livestock raised nearby has higher annual mean PM2.5 concentration. The results are expected to provide the government bodies to make environmental decisions and the plants and livestock farms to install air monitors to analyze the air quality data. Our ultimate goals are to improve the environment and reduce both the emission of PM2.5 and the probability of getting cardiovascular disease.

The subject of air pollution is paid increasing attention to in recent years. NH3 has significant effect on PM2.5, especially due to animal excrements and chemical fertilizer. Some PM2.5 monitoring data in Taiwan show that the concentration in south central Taiwan is apparently on the high side, and south central Taiwan has dense livestock and poultry. Therefore, this paper combines the livestock and poultry data opened by Taiwan Council of Agriculture (COA) with the PM2.5 data obtained by air monitoring stations opened by Taiwan Environmental Protection Administration (EPA), and uses SpatialHadoop to build a Cloud platform to analyze the correlation between Taiwan’s livestock data and the concentration of PM2.5. The analysis results show that the annual mean concentration of PM2.5 of the air monitoring station in the livestock and poultry dense region is higher than that in other regions by 33%.

The aim of this study was to investigate the influence of rice bran oil consumption on plasma lipids and insulin resistance in patients with type 2 diabetes. Thirty-five patients with type 2 diabetes were randomly assigned to a placebo group or a rice bran oil group. The placebo group consumed 250 mL soybean oil-modified milk (18 g soybean oil) daily for 5 weeks, and the rice bran oil group consumed 250 mL rice bran oil modified milk (18 g rice bran oil) daily for 5 weeks. At week 0 and week 5, anthropometric measurements, hematology tests, and an oral-glucose-tolerance test were conducted. The results showed that the homeostasis model assessment index of insulin resistance, the area under the curve for postprandial serum insulin, and serum low-density-lipoprotein cholesterol concentrations increased significantly in the placebo group. In the rice bran oil group, fasting and 2-h postprandial blood glucose concentrations and the area under the curve for postprandial plasma glucose increased significantly; however, total serum cholesterol and low-density-lipoprotein cholesterol concentrations decreased significantly. However, the homeostasis model assessment index of insulin resistance was not significantly different. Consumption of 18 g rice bran oil modified milk daily for 5 weeks significantly decreased total serum cholesterol concentrations and tended to decrease low-density-lipoprotein cholesterol concentrations in patients with type 2 diabetes. However, no significant influence on insulin resistance was observed.

To evaluate the prevalence of obstructive sleep apnea (OSA) and to clarify sleep characteristics in patients with mucopolysaccharidoses (MPS), we performed overnight polysomnographic studies in 24 patients (22 males and 2 females; 3 with MPS I, 15 with MPS II, 1 with MPS III, 1 with MPS IV, and 4 with MPS VI; mean age, 10.8 ± 6.0 years; age range, 2.0-23.7 years; 2 patients ≥18 years of age). The nadir arterial oxygen saturation (SaO(2) ) was 74.5 ± 12.3%, and the average percentage of sleep time with an SaO(2) of95% was 39.4%. The percentages of total sleep time spent in sleep stages N1, N2, N3, and R were 18.6 ± 10.8%, 50.3 ± 7.6%, 14.8 ± 8.1%, and 15.3 ± 4.6%, respectively. The respiratory disturbance index (RDI) was 21.8 ± 20.4/hr, and obstructive apnea-hypopnea index (OAHI) and central apnea index were 21.4 ± 19.9/hr and 0.4 ± 0.6/hr, respectively. The desaturation index was 17.6 ± 17.8/hr. All patients had some degree of OSA. For 22 children, the disorder was mild (OAHI 1.5-5) in 2, moderate (OAHI 5-10) in 7, and severe (OAHI 10) in 13. Two patients with MPS II who received enzyme replacement therapy had reductions in RDI after treatment (38.9-10.8 and 3.5-2.0, respectively). The prevalence of moderate to severe OSA was 88% (21/24) in patients with MPS. The overnight polysomnography will help to determine the abnormalities of breathing during sleep more precisely and urge the clinicians to take necessary action for patients with severe manifestations.

An efficient method is presented to characterize the damage state for two-dimensional multicracked elastic solids. This method is based on the concept of a path-independent Mc -integral, through which the surface energy associated with creation of all the cracks is evaluated. On one hand, when the cracked media are homogeneously stressed, the correspondence relation between the effective material moduli and Mc is established. On the other hand, when the cracked solids are nonhomogeneously stressed, the effective structural stiffness is determined by using the result of Mc . Through proper use of Mc , the damage state of the cracked structure can be assessed both qualitatively and quantitatively.

Purpose: We herein examine whether macrophage inflammatory protein-3α (MIP-3α) is a biomarker for nasopharyngeal carcinoma (NPC) and whether it is involved in modulating NPC cell functions.Experimental Design: The study population comprises 275 NPC patients and 250 controls. MIP-3α levels in tissues and sera were examined by immunohistochemistry and ELISA, respectively. EBV DNA load and EBV viral capsid antigen IgA were measured by quantitative real-time PCR and immunofluorescence assay, respectively. Effects of MIP-3α on NPC cell motility were investigated by Transwell migration/invasion assays and RNA interference.Results: MIP-3α was overexpressed in NPC tumor cells. Serum MIP-3α levels were significantly higher in untreated patients, recurrent patients and patients with distant metastases versus non-NPC controls, patients with complete remission, and long-term disease-free patients. In the prospective cohort, serum MIP-3α levels were significantly higher in untreated NPC patients with advanced tumor-node-metastasis stage versus early stage and also correlated with EBV DNA load. Measurement of MIP-3α, EBV DNA, and viral capsid antigen IgA levels in serial serum/plasma samples from treated patients at 6-month intervals revealed a high association between MIP-3α level, EBV DNA load, and disease status. Among 155 consecutive NPC patients, subjects with pretreated MIP-3α serum levels over 65 pg/mL had worse prognoses for overall survival and distant metastasis-free survival in univariate and multivariate analysis. Additionally, cell functional assays showed that MIP-3α contributed to migration and invasion of NPC cells, which could be effectively inhibited by MIP-3α knockdown.Conclusions: MIP-3α may be a novel biomarker and prognosticator for NPC and is involved in migration and invasion of NPC cells.

This study investigated the effects of β-carotene and canthaxanthin on lipid peroxidation and antioxidative enzyme activities in rats fed a high-cholesterol, high-fat diet. Wistar rats were divided into six groups. Negative control group (group NC) received a high-fat (150 g/kg) diet; cholesterol control group (group CC) received a high-cholesterol (10 g/kg), high-fat diet. The other four groups were fed a high-cholesterol, high-fat diet supplemented with crystal β-carotene (group BC), β-carotene beadlet (group BB), canthaxanthin beadlet (group CX) or α-tocopherol (group AT). Blood and livers were collected for analysis after 6 weeks of feeding. Group BB had significantly lower hepatic thiobarbituric acid reactive substance (TBARS) and conjugated diene concentrations, whereas group CX had a significantly lower plasma TBARS concentration than did group CC. In erythrocytes, glutathione peroxidase activities were significantly greater in groups BC, BB and CX than in group CC. Moreover, compared with group CC, catalase activities were significantly greater in groups BB and CX, and superoxide dismutase (SOD) activity was significantly greater in group BB. In livers, SOD activities were significantly greater in groups BC, BB and CX, and glutathione reductase activities were significantly greater in groups BB and CX than in group CC. Compared with group CC, hepatic retinol and α-tocopherol concentrations were significantly greater in groups BC, BB and CX, whereas plasma and hepatic cholesterol concentrations were significantly lower in group BC. These findings suggest that β-carotene and canthaxanthin altered the pro-oxidation and antioxidation balance and suppressed cholesterol-induced oxidative stressviamodulation of antioxidant system and cholesterol metabolism.

Fucosidosis is a rare lysosomal storage disease caused by a defect of the alpha-L: -fucosidase (FUCA1) gene. Worldwide 26 mutations underlying the disease have been reported. By direct DNA sequencing of exons and flanking introns, homozygous Y126X mutation and Q281R polymorphism were found in a Taiwanese patient with fucosidosis. Upon expressing in COS-7 cells, 97.4% of alpha-L: -fucosidase activity compared with that of the wild-type construct was observed in the cDNA containing Q281R polymorphism. Western blot analysis revealed a 58-kDa precursor and 56-kDa mature forms for cells transfected with wild-type and Q281R enzymes. Using the fluorogenic substrate, the Michaelis constants and maximal velocities of both enzymes were very similar. While no appreciable enzyme activity (0.0%) was observed with Y126X mutation, no apparent decrease in FUCA1 mRNA level was seen with Y126X mutation. The expressed truncated Y126X protein was unstable and largely degraded. The delineation of the molecular defect could serve to complement future prenatal diagnosis for this family when necessary.

Transforming growth factor (TGF-β)/TGF-β receptor signal is known to promote cell migration. Up-regulation of TGF-β in serum/peritoneal fluid and increased levels of pluripotent transcription factor OCT4 in endometriotic tissues are frequently observed in patients with endometriosis. However, the mechanisms underlying how TGF-β/TGF-β receptor and OCT4 affect endometriotic cell migration still remain largely unknown. Therefore, endometriotic tissue with high cell migratory capacity were collected from patients with adenomyotic myometrium (n = 23) and chocolate cyst (n = 24); and endometrial tissue with low cell migratory capacity in normal endometrium or hyperplastic endometrium (n = 8) were collected as the controls. We found the mRNA levels of TGF-β receptor I (TGF-β RI) and OCT4 were significantly higher in the high-migratory ectopic endometriotic tissues than those of the low-migratory normal or hyperplastic endometrium. Positive correlations between TGF-β RI and OCT4, and either TGF-β RI or OCT4 with migration-related genes (SNAIL, SLUG and TWIST) regarding the mRNA levels were observed in human endometriotic tissues. TGF-βI dose-dependently increased the gene and protein levels of OCT4, SNAIL and N-Cadherin (N-CAD) and silencing of endogenous OCT4 significantly suppressed the TGF-βI-induced expressions of N-CAD and SNAIL in primary human endometriotic stromal cells and human endometrial carcinoma cell lines RL95-2 and HEC1A. Furthermore, TGF-βI significantly increased the migration ability of endometriotic cells and silencing of OCT4 dramatically suppressed the TGF-βI-induced cell migration activity evidenced by wound-closure assay, transwell assay, and confocal image of F-actin cellular distribution. In conclusion, the present findings demonstrate that the niche TGF-β plays a critical role in initiating expressions of pluripotent transcription factor OCT4 which may contribute to the ectopic endometrial growth by stimulating endometrial cell migration. These findings would be useful for developing therapeutic strategies targeting TGF-β-OCT4 signaling to prevent endometriosis in the future.

Hunter syndrome (mucopolysaccharidosis type II) is an X-linked recessive lysosomal storage disease caused by a defect of the iduronate-2-sulfatase (IDS) gene. The result is impaired IDS enzyme function.To characterize the biochemical and molecular defects in IDS-deficient patients and their families, we measured IDS enzyme activity by fluorimetric enzyme assay and identified the IDS gene mutations in 14 unrelated Taiwanese patients with varying clinical phenotypes. In addition, haplotype analysis was also performed.Three novel (IVS2+1GC, 1055del12, and G489D) and 7 previously reported (N63K, P228L, K347E, R468Q, R468W, I485R, and 1241delAG) mutations were found. Together R468Q and R468W account for 42.8% mutations found in our patients. Haplotype analysis using IDS flanking markers DXS1113 and DXS1123 revealed that the unrelated R468Q alleles were independent in origin whereas the unrelated R468W alleles are probably of the same origin. The R468Q mutation in patient 1150 and I485R mutation in patient 710 occurred de novo in male meioses. Once the mutation in a family was identified, restriction analysis was also performed for rapid diagnosis of female carriers in 8 families. Leukocyte IDS measurement revealed significantly wide range of IDS activity in normal controls and MPS II carriers (19.2 - 70.6 vs. 8.4 - 26.6 nmol/h/mg cell protein). The average leukocyte IDS activity of normal controls (n=43) was 43.9+/-13.3 nmol/h/mg protein, whereas patients with MPS II (n=14) had5% of mean normal IDS activity (0.9+/-0.6 nmol/h/mg protein), and carriers (n=13) had a mean activity of 17.5 (+/-5.7) nmol/h/mg protein. The mean leukocyte IDS activity in female carriers was less than a half of the normal level.Due to a small overlapping range of normal and carriers, the level of enzyme activity cannot be used alone for carrier detection.

DNA screening for LDL receptor mutations was performed in 170 unrelated hyperlipidemic Chinese patients and two clinically diagnosed familial hypercholesterolemia patients. Two deletions (Del e3-5 and Del e6-8), eight point mutations (W-18X, D69N, R94H, E207K, C308Y, I402T, A410T, and A696G), and two polymorphisms (A370T and I602V) were identified. Of these mutations, C308Y and Del e6-8 were found in homozygosity, and D69N and C308Y were seen in unrelated patients. The effects of mutations on LDL receptor function were characterized in COS-7 cells. The LDL receptor level and activity were close to those of wild type in A696G transfected cells. A novel intermediate protein and reduction of LDL receptor activity were seen in D69N transfected cells. For R94H, E207K, C308Y, I402T, and A410T mutations, only approximately 20-64% of normal receptor activities were seen. Conversely, Del e3-5 and Del e6-8 lead to defective proteins with approximately 0-13% activity. Most of the mutant receptors were localized intracellularly, with a staining pattern resembling that of the endoplasmic reticulum and Golgi apparatus (D69N, R94H, E207K, C308Y, and I402T) or endosome/lysosome (A410T and Del e6-8). Molecular analysis of the LDL receptor gene will clearly identify the cause of the patient's hyperlipidemia and allow appropriate early treatment as well as antenatal and family studies.

Hypoxia inducible factor 2α (HIF-2α) is critical for primordial germ cell (PGC) survival as knockout of HIF-2α (HIF-2α(-/-)) decreases both expression of Oct-4 and PGC number in genital ridge. Hypoxia is known to stabilize HIF-2α protein from proteasomal degradation. However, little is known about the hypoxia-associated endocrinal signaling in HIF-2α expression. The current work demonstrates a role for an endocrine insulin-like growth factor-I receptor (IGF-IR)-PI3K/Akt-mTOR-HIF-2α regulatory loop in the proliferation and Oct-4 maintenance of PGC-like alkaline phosphatase positive mouse germline stem cells (AP(+)GSCs). We found that hypoxia greatly increased the cell proliferation and the levels of nuclear Oct-4/HIF-2α protein of AP(+)GSCs. The hypoxic-AP(+)GSCs presented stronger stemness ability for germ cell differentiation than normoxic, with expressions of c-KIT (differentiation germ cell marker), VASA (differentiation germ cell marker) and SCP3 (meiotic marker) using a renal capsule transplantation assay. Meanwhile, hypoxia significantly increased the expression levels of secreted-IGF-I and IGF-IR. The IGF-I dose dependently increased the HIF-2α expression levels in AP(+)GSCs; and, the inhibition of IGF-IR by RNA interference (shIGF-IR) or LY294002 (PI3K inhibitor)/Rapamycin (mTOR inhibitor) effectively suppressed the IGF-I- and/or hypoxia-induced HIF-2α and Oct-4 expression, suggesting that the IGF-IR and its downstream Akt/mTOR signaling are involved in the IGF-I/hypoxia effects. Additionally, knockdown of HIF-2α dramatically suppressed Oct-4 and IGF-IR protein levels in AP(+)GSC cells. In conclusion, the present study demonstrates a regulatory loop of IGF-IR-PI3K/Akt-mTOR-HIF-2α in proliferation and Oct-4 maintenance of PGC-like AP(+)GSCs under hypoxia. This finding provides insights into the niche endocrinology underlying early germ cell development.

Objective To identify the impact of the pluripotent transcription factor OCT4 in endometrial cell migration and endometriosis. Design The OCT4 expression and cell migration study. Setting Research institution and reproductive medical clinic. Patient(s) Nine subjects with normal endometrium, 3 subjects with normal myometrium, 36 patients with hyperplastic endometrium, and 58 patients with endometriosis. Intervention(s) The expression of OCT4 messenger RNA in normal endometrium, normal myometrium, hyperplastic endometrium, and ectopic endometriotic tissues was analyzed using reverse transcription and quantitative real-time polymerase chain reaction (PCR). The effect of OCT4 expression on the migration activity of the endometrial cells was examined. Main Outcome Measure(s) Reverse transcription and quantitative real-time PCR, Western blotting, and wound closure and transwell assays. Result(s) The expression of OCT4 and NANOG messenger RNA was significantly higher in ectopic endometriotic tissues, compared with that of the normal endometrium, the normal myometrium, and the hyperplastic endometrium. The level of OCT4 messenger RNA in endometriotic tissues was positively correlated with the expression of genes associated with cell migration. Overexpression of the OCT4 protein in primary human endometriotic stromal cells and human RL95-2 and HEC1A endometrial carcinoma cell lines resulted in decreased levels of E-CADHERIN, the increased expression of the VIMENTIN, TWIST, and SLUG proteins, and an increase in the migration activity of endometrial cells in transwell and wound closure assays. Conclusion(s) The transcription of the OCT4 gene is significantly up-regulated in human ectopic endometriotic tissues. The expression of OCT4 may contribute to the pathology of ectopic endometrial growth by stimulating the migration activity of endometrial cells.

Claudins are major tight junction proteins that regulate the integrity and function of tight junctions. Aberrant expression of claudins has been shown in various carcinomas with diverse prognostic implications. However, their expression pattern and prognostic value have not been investigated in nasopharyngeal carcinoma. This is the first study to characterize the expression pattern of claudins 1, 4, and 7 in a cohort of 176 patients with nasopharyngeal carcinoma and a minimum follow-up of 9.4 years. Immunohistochemistry with semiquantitative assessment of expression of claudins 1, 4, and 7 was performed on 176 biopsy specimens. Results were correlated with sex, age, extent of tumor, lymph node status, the presence or absence of distant metastasis, and patient survival. High expression of claudins 1 and 4 (72.2% and 88.1%) and low expression of claudin 7 (82.4%) were frequently detected. Low claudin 4 expression (P = .020) and high claudin 7 expression (P < .001) were associated with distant metastasis. Elevated claudin 7 expression also correlated with high tumor stage (P = .001). Furthermore, decreased claudin 4 expression (P = .003) and increased claudin 7 expression (P < .001) independently predicted shorter distant metastasis-free survival by Cox regression analysis. Claudin 4 and claudin 7 may be a novel biomarker for the prediction of distant metastasis and unfavorable prognosis in nasopharyngeal carcinoma.

Nearly 300 different mutations underlying mucopolysaccharidosis type II (MPS II) have been identified worldwide. To investigate the molecular lesions underlying Taiwanese MPS II, probands and families were identified and screened for iduronate-2-sulfatase (IDS) mutation by single-strand conformation polymorphism and DNA sequencing. Five novel and five previously reported mutations were found. Together with those previously reported, a total of 17 identified missense, small deletion, and nonsense mutations were further characterized by transient expression studies. Transfection of COS-7 cells by the mutated cDNA did not yield active enzyme, demonstrating the deleterious nature of the mutations. A 57% decrease in IDS mRNA level was seen with the 231del6 mutation. Among the 11 missense mutations examined, K347E substitution showed apparent normal maturation and targeting on immunoblot and confocal fluorescence microscopy examination. The other 10 missense mutations showed apparent normal precursor with little or reduced mature forms, indicating normal maturation but incorrect targeting of the mutant enzymes. Among the six deletion and nonsense mutations examined, 1055del12 and E521X showed abnormal maturation. The staining pattern of the truncated W267X and 1184delG proteins suggested retention within early vacuolar compartments. The mutated 231del6 and 1421delAG proteins were unstable and largely degraded. Molecular analysis of the IDS gene will clearly identify the cause of the disease within patients and allow antenatal and family studies. The further characterization of gene mutations may delineate their functional consequences on IDS activity and processing and may enable future studies of genotype–phenotype correlation to estimate a prognosis and to lead to possible therapeutic interventions.

Endometriosis is the eutopic and/or ectopic growth of endometrial tissues. High levels of TGF-β in peritoneal fluid and increased expression of OCT4 have been well documented in endometriosis. However, the molecular mechanism between TGF-β and OCT4 expression in endometriosis still remains largely unknown. In this study, we examined the roles of OCT4 in TGF-β-induced endometrial cell migration and the progression of human ectopic endometriosis. We analyzed the plasma levels of TGF-β (n = 80) and gene expression levels of TGF-βR1 in patient tissues (n = 60) by an ELISA assay and quantitative real-time RT-PCR. The serum TGF-β and tissue TGF-βR1 expressions were significantly increased in the ectopic endometriotic tissues (n = 54, 29 for adenomyosis tissues and 25 for chocolate cysts) comparing with the normal endometrium (n = 5). The TGFβR1 transcriptional level in endometriotic tissues was positively correlated with expressions of OCT4 and genes associated with cell migration, such as VIMENTIN, TWIST, SNAIL and SLUG. TGF-β dose-dependently increased the OCT4 expression both in gene and protein levels in human endometriotic stromal cells and, at a less intensity, in HEC1A endometrial carcinoma cell line. Additionally, TGF-βincreased the expression of migration-associated genes and proteins (SNAIL and N-CAD) dose-dependently both in gene and protein levels in human endometriotic stromal cells. Knockdown of OCT4 significantly suppressed the TGF-β-induced the expressions of migration-associated genes and proteins (N-CAD and SNAIL), and promoted the migration of endometrial cells as evidenced by wound-closure and transwell assays. Conclusions: TGF-β,TGF-βRI, and OCT4 are significantly upregulated in human ectopic endometriotic tissues. The significant OCT4 expression may be involved in the pathology of ectopic endometrial growth through TGF-β-induced migration of endometrial cells. Citation Format: Te-Sheng Chang, Heng-Kien Au, Thai-Yen Ling, Ching-Chi Chi, Chii-Ruey Tzeng, Jui-Hung Chang, Yen-Hua Huang. TGF-β1 enhanced cell migration involving pluripotent transcription factor OCT4 in endometriosis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5580. doi:10.1158/1538-7445.AM2014-5580

Expression of cancer stemness in hepatocellular carcinoma (HCC) is linked to drug resistance and tumor relapse which challenge the current therapy. However, mechanisms underlying the regulation of cancer stemness in HCC remain unclear. Effects of interleukin (IL)-6 and insulin-like growth factor I (IGF-I) on stemness expression in HCC were examined using OCT4/NANOG promoter luciferase reporter assays, RNA interference, secondary sphere formation, side population assay, and a xenograft animal model. Western blot analysis and immunohistochemical staining were used to detect the expression of OCT4/NANOG and phospho-IGF-I receptor (p-IGF-IR) in tissues. Levels of OCT4/NANOG and IGF-IR expressions were analyzed by real-time quantitative reverse transcriptase polymerase chain reaction (qPCR) and their association with tumor recurrence of hepatectomized HCC patients (n = 119) was determined using a Kaplan-Meier survival analysis. A high positive correlation between OCT4/NANOG and IGF-IR/p-IGF-IR expressions in human HCC tissues was observed. The concurrent expression of OCT4/NANOG/IGF-IR was mostly confined to HBV-related HCC (HBV-HCC) and was significantly associated with early tumor recurrence. Investigation of the underlying mechanism showed that IL-6 stimulated the autocrinal IGF-I/IGF-IR expression in a STAT dependent manner, and IGF-I/IGF-IR activation promoted the expression of stemness-related properties of Hep3B cells in both cell lines and xenograft murine models. IL-6 stimulated the expression of stemness-related properties in Hep3B cells, and inhibition of IGF-IR activation significantly suppressed the IL-6’s effects both in vitro and in vivo. Conclusions: Expression of OCT4/NANOG is regulated by IL-6-mediated IGF/IGF-IR signaling in HBV-HCC and is associated with the early recurrence of HCC. Citation Format: Te-Sheng Chang, Thai-Yen Ling, Yu-Chih Wu, Wei-Chi Su, Jaw-Ching Wu, Ching-Chi Chi, Pey-Jium Chang, Jui-Hung Chang, Ya-Ting Yang, Kam-Fai Lee, Jun-Jen Liu, Shui-Yi Tung, Yen-Hua Huang, Liang-Mou Kuo, Hong-Nerng Ho, Yen-Hua Huang. IL-6 and IGF-1 receptor signal in stemness expression of HBV-related hepatocellular carcinoma . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1065. doi:10.1158/1538-7445.AM2013-1065