Identification of Blood Exosomal miRNA-1246, miRNA-150-5p, miRNA-5787 and miRNA-8069 as Sensitive Biomarkers for Hepatitis B Virus Infection.

BACKGROUND: There is a big difference in the expression of miRNAs of plasma exosomes of patients with HBV infection. This study aims to analyze four molecular markers of peripheral blood plasma exosomes to evaluate their potential diagnostic values in HBV infection. METHODS: A total of 55 cases of patients with chronic hepatitis B were in Experimental Group 1; 49 cases of hepatitis B carriers were in Experimental Group 2, and 46 cases were in the healthy control group. The total RNA of the plasma exosome was used to analyze the specificity and sensitivity and draw ROC curves. RESULTS: There was a significant difference in the expression of miRNA-1246, miRNA-150-5p, miRNA-5787, and miRNA-8069 down-regulated by plasma exosomes in Experimental Group 1 and Group 2 and Control Group, with a p value of less than 0.05. CONCLUSIONS: The molecular markers down-regulated were miRNA-1246, miRNA-150-5p, miRNA-5787, and miRNA-8069. The four miRNAs were initially identified as new markers of miRNAs of peripheral blood plasma exosomes after HBV infection. It is better to use multiple markers for combined diagnosis.

Diagnostic performance of an immunoassay based on urine exfoliated cell enrichment nanotechnology for upper tract urothelial carcinoma: a retrospective, monocentric study.

BACKGROUND: Noninvasively urine-based diagnostic modalities for upper urinary tract urothelial carcinoma (UTUC) were still lacking. We evaluated the diagnostic value of our previously developed urine-based assay (UTC assay) in UTUC. METHODS: We retrospectively analyzed 90 patients with suspected UTUC and 40 donors without UTUC. Voided urine specimens were analyzed by UTC assay and fluorescence in situ hybridization (FISH). The performance of UTC assay and FISH was compared among the 60 histologically proven UTUC patients and the 40 donors with benign disease. RESULTS: Of the 60 UTUCs, there were 8 low-grade and 52 high-grade cases. Overall sensitivity for UTC assay and FISH were 85% and 73.3%, respectively (P = 0.116). Specificities for UTC assay and FISH were 92.5% and 95%, respectively (P = ns.). By grade, sensitivities of UTC assay and FISH were 87.5% vs. 37.5% for low-grade (P = 0.119), and 84.6% vs. 78.8% for high- grade UTUC (P = 0.446), respectively. By stage, UTC assay showed significantly higher sensitivity than FISH for detecting non-muscle-invasive UTUC, which were 88.5% vs. 61.5%, respectively (P = 0.025). CONCLUSION: UTC assay has good performance for the non-invasive diagnosis of UTUC. UTC assay may improve the diagnosis and surveillance of low-grade or superficial UTUC.

A novel diagnostic nomogram based on serological and ultrasound findings for preoperative prediction of malignancy in patients with ovarian masses.

OBJECTIVE: To develop a novel diagnostic nomogram model to predict malignancy in patients with ovarian masses. METHODS: In total, 1277 patients with ovarian masses were retrospectively analyzed. Receiver operating characteristic (ROC) analysis was performed to identify valuable predictive factors. Univariate and multivariate logistic regression analyses were used to identify risk factors for ovarian cancer. Subsequently, a predictive nomogram model was developed. The performance of the nomogram model was assessed by its calibration and discrimination in a validation cohort. Decision curve analysis (DCA) was applied to assess the clinical net benefit of the model. RESULTS: Overall, 496 patients (38.8%) had ovarian cancer. Eighteen parameters were significantly different between the malignant and benign groups. Five parameters were identified as being most optimal for predicting malignancy, including age, carbohydrate antigen 125, fibrinogen-to-albumin ratio, monocyte-to-lymphocyte ratio, and ultrasound result. These parameters were incorporated to establish a nomogram model, and this model exhibited an area under the ROC curve (AUC) of 0.937 (95% confidence interval [CI], 0.920-0.954). The model was also well calibrated in the validation cohort and showed an AUC of 0.925 (95%CI, 0.896-0.953) at the cut-off point of 0.298. DCA confirmed that the nomogram model achieved the best clinical utility with almost the entire range of threshold probabilities. The model has demonstrated superior efficacy in predicting malignancy compared to currently available models, including the risk of ovarian malignancy algorithm, copenhagen index, and the risk of malignancy index. More importantly, the nomogram established here showed potential value in identification of early-stage ovarian cancer. CONCLUSION: The cost-effective and easily accessible nomogram model exhibited favorable accuracy for preoperative prediction of malignancy in patients with ovarian masses, even at early stages.

High-performance sulphate-free cleansers: Surface activity, foaming and rheology.

OBJECTIVE: The main objective of this paper is to analyse the composition of a sulphate-free binary or ternary surfactant system with alkyl olefin sulfonate (AOS), alkyl polyglucoside (APG) and lauryl hydroxysultaine (Sultaine). The composition was optimized by observing critical parameters such as surface activity and rheological properties while varying the concentration of APG in a ternary system, varying the ratio of AOS and Sultaine in a binary system and studying the effect of sodium chloride addition. The experimental results can provide an alternative, sulphate-free surfactant system to replace the common system containing sodium laureth sulfate (SLES) and cocamidopropyl betaine (CapB), without compromising on the parameters previously mentioned. METHODS: A DuNouy Ring was utilized on a tensiometer to measure the surface tension of the samples. To observe foaming abilities of samples, a visual foaming study was conducted and recorded by taking pictures. A TA instruments mechanical rheometer was used to measure the viscosity. RESULTS: Studying the effect of APG concentration on surface tension illustrated that as APG concentration decreases, surface tension decreases as well. The minimum surface tension was found to be 26.587 for 7.5 wt.% AOS and 7.5 wt.% Sultaine. When the ratio between AOS and Sultaine changed, the 1:1 system produced the lowest surface tension value again. As the concentration of AOS decreased in the ratio, the surface tension increased. When the ratio was held constant, and APG was introduced into the system, the systems containing APG had higher surface tension values compared with the systems with the same ratios but did not have APG added. As the concentration of salt increased, the surface tension decreased for AOS, increased for the binary mixture and had no effect on Sultaine. Foaming has a direct correlation with surface tension so a decrease in surface tension led to better foaming abilities; therefore, the 1:1 ratio of AOS:Sultaine had the best foaming qualities. The AOS/Sultaine system exhibited Maxwellian behaviour, suggesting the presence of worm-like micellar structures. As the concentration of APG increased, the viscosity decreased and at a concentration of 2.5 wt.% AOS, 5 wt.% APG and 7.5 wt.% Sultaine, the system changed from shear thinning non-Newtonian fluid to Newtonian. Varying the ratio of AOS:Sultaine in the binary system, showed as the ratio changed, the viscosity decreased, and at a ratio of 1:3 AOS:Sultaine, the system was primarily Newtonian. As the ratio remained constant and APG was added, the viscosity continued to decrease and the 1:3 ratio was completely Newtonian. The addition of salt had no effect on the viscosity of Sultaine, but the viscosity of AOS increased at a salt concentration of 2 wt.%. The binary mixture saw as the concentration of salt increased, the viscosity profile decreased, even though the system remained non-Newtonian and shear thinning. CONCLUSION: The surface activity and rheological study of the sulfate-free surfactant systems unveil a strong synergistic interaction between AOS and Sultaine specifically at a ratio of 1:1 resulting in high surface activity and corresponding good foaming and formation of entangled wormlike micelles resulting in excellent viscosity build in the system. If introducing a biobased surfactant into this system is desired without completely compromising the performance properties, the ternary surfactant system 4.5 wt.% AOS, 3 wt.% APG and 7.5 wt.% Sultaine should be considered optimal. Any further addition of APG or changing the ratio results in detrimental reductions of all performance controlling physio-chemical parameters.

OBJECTIF: l'objectif principal de cet article est d'analyser la composition d'un système de surfactant binaire ou ternaire sans sulfate utilisant l'alkyle sulfonate oléfine (alkyl olefin sulfonate, AOS), l'alkyle polyglucoside (APG) et le lauryl hydroxysultaïne (sultaïne). La composition a été optimisée en observant des paramètres critiques tels que l'activité de surface et les propriétés rhéologiques tout en faisant varier la concentration d'APG dans un système ternaire, le rapport d'AOS et de sultaïne dans un système binaire et en étudiant l'effet de l'ajout de chlorure de sodium. Les résultats expérimentaux peuvent fournir un système de surfactant alternatif sans sulfate pour remplacer le système courant contenant le laureth sulfate de sodium (sodium laureth sulfate, LES) et de la camidopropyle bétaïne (cocamidopropyl betaine, CapB), sans compromettre les paramètres précédemment mentionnés. MÉTHODES: un anneau de DuNouy a été utilisé sur un tensiomètre pour mesurer la tension superficielle des échantillons. Pour observer les capacités de formation de mousse des échantillons, une étude visuelle de formation de mousse a été menée et les résultats enregistrés en prenant des photos. Un rhéomètre mécanique TA Instruments a été utilisé pour mesurer la viscosité. RÉSULTATS: l'étude de l'effet de la concentration d'APG sur la tension superficielle a illustré qu'à mesure que la concentration d'APG diminue, la tension superficielle diminue également. La tension superficielle minimale s'est avérée être de 26,587 pour une concentration d'AOS de 7,5 % en poids et de sultaïne de 7,5 % en poids. Lorsque le rapport entre l'AOS et la sultaïne a été modifié, le système à un rapport de 1:1 a de nouveau produit la valeur de tension superficielle la plus basse. À mesure que la concentration d'AOS diminuait dans le rapport, la tension superficielle augmentait. Lorsque le rapport a été maintenu constant, et que l'APG a été introduit dans le système, les systèmes contenant l'APG présentaient des valeurs de tension superficielle plus élevées que les systèmes ayant les mêmes rapports, mais n'ont pas fait l'objet d'ajout d'APG. À mesure que la concentration de sel augmentait, la tension superficielle diminuait pour l'AOS, augmentait pour le mélange binaire et n'avait aucun effet sur la sultaïne. La formation de mousse est directement corrélée à la tension superficielle, de sorte qu'une diminution de la tension superficielle a entraîné de meilleures capacités de formation de mousse ; par conséquent, le rapport AOS/sultaïne de 1:1 présentait les meilleures qualités de formation de mousse. Le système AOS/sultaïne a montré un comportement maxwellien, suggérant la présence de structures micellaires en forme de ver. À mesure que la concentration d'APG augmentait, la viscosité diminuait, et à une concentration d'AOS de 2,5 % en poids, d'APG de 5 % en poids et de sultaïne de 7,5 % en poids, le système est passé d'un fluide non Newtonien par cisaillement (Rhéofluidification) à un fluide Newtonien. En variant le rapport d'AOS/sultaïne dans le système binaire, on a montré que le rapport changeait, la viscosité diminuait et, à un rapport d'AOS/sultaïne de 1:3, le système était principalement Newtonien. Lorsque le rapport est resté constant et que l'APG a été ajouté, la viscosité a continué à diminuer et le rapport 1:3 était complètement Newtonien. L'ajout de sel n'a eu aucun effet sur la viscosité de la sultaïne, mais la viscosité de l'AOS a augmenté à une concentration en sel de 2 % en poids. Le mélange binaire montrait qu'à mesure que la concentration de sel augmentait, le profil de viscosité diminuait, même si le système restait non Newtonien par cisaillement (Rhéofluidification). CONCLUSION: l'étude de l'activité de surface et rhéologique des systèmes de surfactant sans sulfate dévoile une forte interaction synergique entre l'AOS et la sultaïne, spécifiquement à un rapport de 1:1, qui entraîne une activité de surface élevée et une bonne formation correspondante de mousse et de micelles enchevêtrées, ce qui aboutit à une excellente formation de viscosité dans le système. Si on souhaite introduire un surfactant biosourcé dans ce système sans compromettre complètement les propriétés de performance, le système de surfactant ternaire à une concentration d'AOS de 4,5 % en poids, d'APG de 3 % en poids et de sultaïne de 7,5 % en poids doit être considéré comme optimal. Tout ajout supplémentaire d'APG ou toute modification du rapport entraîne des réductions préjudiciables de toutes les performances contrôlant les paramètres physicochimiques.

Prevalence, serotypes, and antimicrobial resistance of Salmonella isolates from patients with diarrhea in Shenzhen, China.

BACKGROUND: Salmonella is one of the main causative agents of diarrhea which results in substantial disease burden. To determine the prevalence, serotype distribution, and antimicrobial resistance profiles of clinical Salmonella isolates in Shenzhen, a 6-year surveillance study was conducted. RESULTS: A total of 297 (5.7%) Salmonella strains were isolated from stool samples from 5239 patients. Among the 42 serotypes identified, serotype Typhimurium was the most common one which represented 39.7% of the isolates (118), followed by serotype Enteritidis (71, 23.9%), London (12, 4.0%), 4, 5, 12: i: - (11, 3.7%), and Senftenberg (8, 2.7%). A high frequency of resistance was found in ampicillin (70.6%), piperacillin (64.5%), tetracycline (63.5%), and streptomycin (54.3%). Resistance to ampicillin and tetracycline was observed in 95.3% of S. Typhimurium isolates; and nalidixic acid in 93.1% of S. Enteritidis isolates. Resistance to 5 or more antimicrobial agents was found in 78.8% of S. Typhimurium and 69.0% of S. Enteritidis isolates. A decreased susceptibility to ciprofloxacin and levofloxacin was associated with amino acid alteration in gyrA gene. Point mutations without amino acid changes were seen in gyrB, parC, and parE genes. CONCLUSIONS: A broad range of serotypes are responsible for Salmonellosis in Shenzhen, with Enteritidis and Typhimurium being the most common serotypes. The high level of antibiotic resistance is of public health significance and ongoing monitoring combined with rational use of antibiotics are recommended. Point mutations in gyrA gene might play an important role in the resistance to fluoroquinolones.

Gene set based systematic analysis of prostate cancer and its subtypes.

Aim: A gene set based systematic analysis strategy is used to investigate prostate tumors and its subclusters with focuses on similarities and differences of biological functions. Results: Dysregulation of methylation status, as well as RAS/RAF/ERK and PI3K-ATK signaling pathways, were found to be the most dramatic changes during prostate cancer tumorigenesis. Besides, neural and inflammation microenvironment is also significantly divergent between tumor and adjacent tissues. Insights of subclasses within prostate tumor cohorts revealed four different clusters with distinct gene expression patterns. We found that samples are mainly clustered by immune environments and proliferation traits. Conclusion: The findings of this article may help to advance the progress of identifying better diagnosis biomarkers and therapeutic targets.

Lipid Production by Rhodotorula glutinis in Continuous Cultivation with a Gravity Sedimentation System.

Lipid accumulation is generally believed to be a partially growth-coupled biochemical process that results in differences in lipid content between different cells. To separate lipid-rich cells and increase the cellular biomass in bioreactors during the cultivation of the oleaginous yeasts, a gravity sedimentation system (GSS) is coupled to a bioreactor. The dilution rate (D) and the ratio of the outflow rate from the outlet of the GSS to the inflow rate into the bioreactor (B) were evaluated. The biomass in the bioreactor with GSS increased by 16.3% and 30.6% at D values of 0.05 h-1 (B = 0.25) and 0.02 h-1 (B = 0.5), respectively. Interestingly, cells containing 39.3% lipids were obtained from the outlet of the GSS (D = 0.02 h-1, B = 0.5), and the lipid content increased by 7.8% compared to that of the bioreactor. The results indicated that use of a GSS is a very effective method for increasing the cell concentration and separation of lipid-rich cells.

Clinical significance of circulating tumor cells in predicting disease progression and chemotherapy resistance in patients with gestational choriocarcinoma.

Gestational choriocarcinoma (GC) is a highly aggressive tumor. In our study, we systematically investigated EpCAM/CD147 expression characteristics in patients with GC and assessed the role of circulating tumor cells (CTCs) in predicting chemotherapy response and disease progression. GC tissues were positive for either epithelial cellular adhesion molecule (EpCAM) or CD147, and all samples exhibited strong human chorionic gonadotropin (HCG) expression. Among all the recruited patients (n = 115), 103 had at least 1 CTC in a 7.5-mL peripheral blood sample, and the percentage of patients with ≥4 CTCs in a particular FIGO stage group increased with a higher FIGO stage (p < 0.001). Furthermore, the pretreatment CTC count was related to tumor size (r = 0.225, p = 0.015) and the number of metastases (r = 0.603, p < 0.001). A progression analysis showed that among the 115 included patients who qualified for further examination, 52 of the 64 patients defined as progressive had ≥4 pretreatment CTCs, while only 7 of the 51 non-progressive patients had ≥4 pretreatment CTCs (p < 0.001). In multivariate analysis, CTCs (≥4) remained the strongest predictor of PFS when other prognostic markers, FIGO score and FIGO stage were included. Moreover, based on the chemotherapy response, patients with ≥4 CTCs were more likely to be resistant to chemotherapy than those with <4 CTCs (P < 0.001). These findings demonstrates the feasibility of CTC detection in cases of GC by adopting EpCAM/CD147 antibodies together as capturing antibodies. The CTC count is a promising indicator in the evaluation of biological activities and the chemotherapy response in GC patients.

Two different pathogenic gene mutations coexisted in the same hereditary spherocytosis family manifested with heterogeneous phenotypes.

BACKGROUND: Hereditary spherocytosis (HS) is a common type of hereditary hemolytic anemia. According to the current diagnostic criteria of HS, patients with a family history of HS, typical clinical features and laboratory investigations could be diagnosed without the requirement of any additional tests, including genetic analysis. However, the clinical heterogeneities incur difficulties in HS diagnosis. We therefore aimed to investigate the application of genetic diagnosis in a family-based cohort. CASE PRESENTATION: In the present Chinese family, two probands sharing similar clinical manifestations, including jaundice, cholelithiasis, splenomegaly and spherocytes, while the clinical features of other family members were inconclusive. Whole-exome sequencing (WES) unexpectedly unveiled two separate disease-causing mutations in the two probands. SPTB R1625X mutation detected in proband D was a de novo mutation; while proband W inherited the SLC4A1 c.G1469A mutation from her mother, which was also inherited by her brother. However, the clinical features of proband W and her mother and brother were discrepant: proband W suffered from significant splenomegaly, jaundice and cholelithiasis, which resulted in cholecystectomy and splenectomy; while her mother and brother's HS were not complicated by cholelithiasis, and their splenomegaly and elevated serum bilirubin were moderate. In addition, additional genomic defects involved with HS-related symptoms have not been detected in this family. CONCLUSIONS: Both genotypes and phenotypes could be heterogeneous in the same HS family. The analysis of pathogenic gene mutations may endeavor to play an indispensable role in the accurate diagnosis and genetic consultation of HS individuals and their family members.

The etiology of acute meningitis and encephalitis syndromes in a sentinel pediatric hospital, Shenzhen, China.

BACKGROUND: Acute meningitis and encephalitis syndromes (AMES) is a severe neurological infection which causes high case fatality and severe sequelae in children. To determine the etiology of childhood AMES in Shenzhen, a hospital-based study was undertaken. METHODS: A total of 240 cerebrospinal fluid (CSF) samples from 171 children meeting the case definition were included and screened for 12 common causative organisms. The clinical data and conventional testing results were collected and analyzed. Whole genome sequencing was performed on a Neisseria meningitidis isolate. RESULTS: A pathogen was found in 85 (49.7%) cases; Group B Streptococcus (GBS) was detected in 17 cases, Escherichia coli in 15, Streptococcus pneumoniae in 14, enterovirus (EV) in 13, herpes simplex virus (HSV) in 3, N. meningitidis in 1, Haemophilus influenzae in 1, and others in 23. Notably, HSV was found after 43 days of treatment. Twelve GBS and 6 E. coli meningitis were found in neonates aged less than 1 month; 13 pneumococcal meningitis in children aged > 3 months; and 12 EV infections in children aged > 1 year old. The multilocus sequence typing of serogroup B N. meningitidis isolate was ST-3200/CC4821. High resistance rate to tetracycline (75%), penicillin (75%), and trimethoprim/sulfamethoxazole (75%) was found in 4 of S. pneumoniae isolates; clindamycin (100%) and tetracycline (100%) in 9 of GBS; and ampicillin (75%) and trimethoprim/sulfamethoxazole (67%) in 12 of E. coli. CONCLUSIONS: The prevalence of N. meningitidis and JEV was very low and the cases of childhood AMES were mainly caused by other pathogens. GBS and E. coli were the main causative organisms in neonates, while S. pneumoniae and EV were mainly found in older children. HSV could be persistently found in the CSF samples despite of the treatment. A better prevention strategy for GBS, the introduction of pneumococcal vaccine, and incorporation of PCR methods were recommended.

Overexpression of SLC12A5 is associated with tumor progression and poor survival in ovarian carcinoma.

INTRODUCTION: The solute carrier family 12 member 5 (SLC12A5) gene is playing a putative oncogenic role in colorectal carcinoma. However, the status of SLC12A5 amplification and expression in ovarian carcinoma and its potential clinical and/or prognostic significance has not yet been investigated. METHODS: In the present study, semi-quantitative staining and fluorescence in situ hybridization were used to investigate SLC12A5 protein expression and gene amplification levels. Samples were obtained from archival, formalin-fixed, paraffin-embedded pathological specimens consisting of 30 normal ovaries, 30 ovarian cystadenomas, 30 borderline ovarian tumors, and 147 invasive ovarian carcinomas. SLC12A5 immunohistochemical staining results, pathological parameters, and patient prognosis were then evaluated using various statistical models. Patient survival rate was also assessed using receiver-operator curve analysis. RESULTS: Our results revealed no SLC12A5 protein overexpression in normal ovaries. However, 7% of cystadenomas had SLC12A5 protein overexpression along with 17% of borderline tumors and 37% of ovarian carcinomas (P<0.01). Amplification of SLC12A5 was detected in 10.3% of ovarian carcinomas. Further correlational analyses showed that SLC12A5 protein overexpression in ovarian carcinomas was significantly associated with ascending histological grade, pT/pN/pM status, as well as FIGO stage (P<0.05). A subsequent univariate survival analysis of our ovarian carcinoma cohorts resulted in a significant association between SLC12A5 protein overexpression and decreased patient survival (44.3 and 85.9 months for high and low SLC12A5 protein expression, respectively; P<0.001). Importantly, additional multivariate analysis revealed that SLC12A5 protein expression was a significant, independent prognostic factor for overall survival in ovarian carcinoma patients (P=0.003). CONCLUSIONS: Collectively, these findings support the conclusion that SLC12A5 protein overexpression could indicate an invasive and/or aggressive phenotype of ovarian carcinoma. Future work will need to investigate whether SLC12A5 protein can serve as an independent prognostic molecular marker in patients with ovarian carcinoma.

[Effects of fungi fraction on growth and anti-oxidative activity of Eleutherococcus senticosus].

The present study was conducted to explore the effect of endophytic fungi fraction on growth and anti-oxidative activity of Eleutherococcus senticosus. The growth,yield,contents of MDA,and antioxidant activities were assessed in E. senticosus under five fungi fractions,namely BZ,MH,DT,JS,and XFZ. The results showed that fungi fractions and component significantly affected the growth,low concentration of DT fungi fraction significantly increased the biomass of E. senticosus,reduced the MDA content in cells,and the antioxidant activities of the aqueous extracts were superior to the others. The results indicated that low concentration of DT fungi fraction was the optimum fraction to achieve high yield and quality of E. senticosus.

Expression of TBC1D16 Is Associated with Favorable Prognosis of Epithelial Ovarian Cancer.

Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy with high recurrence and poor prognosis duo to the lack of effective biomarkers. TBC1 domain family member 16 (TBC1D16), a GTPase-activating protein, is involved in regulating intracellular trafficking in tumorigenesis and metastasis. However, the clinical significance of TBC1D16 in EOC remains unknown. In the present study, we investigated the expression and prognostic significance of TBC1D16 in EOC and its relationship with the expression of vascular endothelial growth factor (VEGF). The tissue specimens included 156 histologically confirmed EOC and 30 normal ovarian tissues. The expression of TBC1D16 and VEGF was detected by immunohistochemistry (IHC), and the immunoreactive score was calculated with signal intensity and percentage of positive cells. IHC results showed that TBC1D16 and VEGF were both mainly localized in cytoplasm of epithelial cells in normal ovarian tissues and were expressed in cancer cells. Based on the immunoreactive score, TBC1D16 expression in EOC was categorized as "high expression," compared with normal ovarian tissues (P < 0.05). The Chi-square test showed that high TBC1D16 expression was related to advanced pT stages (P = 0.029), but not correlated with other clinical features. Moreover, the TBC1D16 expression was significantly higher in EOC specimens with low VEGF expression (P < 0.001). Importantly, in both univariate and multivariate survival analyses, high expression of TBC1D16 was significantly correlated with good overall survival (OS). In conclusion, TBC1D16 is a predictive marker for favorable prognosis of EOC.

Compositional profiles of Rhodosporidium toruloides cells under nutrient limitation.

Lipid production by the red yeast Rhodosporidium toruloides was explored under nutrient limitation. To determine the compositional profiles of R. toruloides cells, samples were prepared using a continuous cultivation process under nutrient limitation and analyzed via several methods, including Fourier transform infrared spectroscopy and elemental analysis. Under nitrogen limitation, as the dilution rate increased, the cellular lipid content decreased but the carbohydrate and protein contents increased. Under carbon limitation, the cellular lipid, protein, and carbohydrate contents remained relatively constant at the different dilution rates. Moreover, the cellular elemental composition was essentially identical under nitrogen and carbon limitation at a high dilution rate of 0.20 h-1. We also analyzed the consumed carbon to nitrogen (C/N) under different nutrition conditions. The results indicated that the consumed C/N had a major influence on cell metabolism and product formation, which contributed to our understanding of the physiological characteristics of R. toruloides.

Dynamics of the lipid droplet proteome of the Oleaginous yeast rhodosporidium toruloides.

Lipid droplets (LDs) are ubiquitous organelles that serve as a neutral lipid reservoir and a hub for lipid metabolism. Manipulating LD formation, evolution, and mobilization in oleaginous species may lead to the production of fatty acid-derived biofuels and chemicals. However, key factors regulating LD dynamics remain poorly characterized. Here we purified the LDs and identified LD-associated proteins from cells of the lipid-producing yeast Rhodosporidium toruloides cultured under nutrient-rich, nitrogen-limited, and phosphorus-limited conditions. The LD proteome consisted of 226 proteins, many of which are involved in lipid metabolism and LD formation and evolution. Further analysis of our previous comparative transcriptome and proteome data sets indicated that the transcription level of 85 genes and protein abundance of 77 proteins changed under nutrient-limited conditions. Such changes were highly relevant to lipid accumulation and partially confirmed by reverse transcription-quantitative PCR. We demonstrated that the major LD structure protein Ldp1 is an LD marker protein being upregulated in lipid-rich cells. When overexpressed in Saccharomyces cerevisiae, Ldp1 localized on the LD surface and facilitated giant LD formation, suggesting that Ldp1 plays an important role in controlling LD dynamics. Our results significantly advance the understanding of the molecular basis of lipid overproduction and storage in oleaginous yeasts and will be valuable for the development of superior lipid producers.

Combined mutagenesis of Rhodosporidium toruloides for improved production of carotenoids and lipids.

OBJECTIVES: To improve production of lipids and carotenoids by the oleaginous yeast Rhodosporidium toruloides by screening mutant strains. RESULTS: Upon physical mutagenesis of the haploid strain R. toruloides np11 with an atmospheric and room temperature plasma method followed by chemical mutagenesis with nitrosoguanidine, a mutant strain, R. toruloides XR-2, formed dark-red colonies on a screening plate. When cultivated in nitrogen-limited media, XR-2 cells grew slower but accumulated 0.23 g lipids/g cell dry wt and 0.75 mg carotenoids/g CDW. To improve its production capacity, different amino acids and vitamins were supplemented. p-Aminobenzoic acid and tryptophan had beneficial effects on cell growth. When cultivated in nitrogen-limited media in the presence of selected vitamins, XR-2 accumulated 0.41 g lipids/g CDW and 0.69 mg carotenoids/g CDW. CONCLUSIONS: A mutant R. toruloides strain with improved production profiles for lipids and carotenoids was obtained, indicating its potential to use combined mutagenesis for a more productive phenotype.

CPE overexpression is correlated with pelvic lymph node metastasis and poor prognosis in patients with early-stage cervical cancer.

PURPOSE: Elevated carboxypeptidase E (CPE) levels play crucial roles in tumorigenesis and metastasis. This study investigated the expression and clinicopathological significance of CPE in early-stage cervical cancer. METHODS: Elevated carboxypeptidase E expression was analyzed using quantitative polymerase chain reaction and western blotting in normal cervical tissue, cervical cancer cell lines, and in cervical cancer tissues and adjacent noncancerous tissues (ANTs) from the same patient. Immunohistochemistry (IHC) was used to examine CPE expression in tissue samples from 112 patients with early-stage cervical cancer (FIGO stages Ia2-IIa2), 60 patients with cervical intraepithelial neoplasia, and 19 patients with normal cervical tissues (NCTs). Associations between CPE expression and prognostic and diagnostic factors were evaluated statistically. RESULTS: CPE expression was significantly higher in cervical cancer cell lines and tissues than in normal tissues and ANTs. Semi-quantitative analysis of IHC indicated that CPE gradually increased from CIN I to cervical cancer, but was absent in NCTs. CPE expression was seen in 40.2 % (45/112) of the cervical cancer samples. CPE expression was significantly associated with FIGO stage (P = 0.003), tumor size (P = 0.012), stromal invasion (P < 0.001), lymphovascular space invasion (P = 0.016), parametrial infiltration (P = 0.027), vaginal involvement (P = 0.007), postoperative adjuvant therapy (P = 0.024), recurrence (P < 0.001), survival (P < 0.001), and pelvic lymph node metastasis (PLNM) (P < 0.001), and it was significantly higher in tissues from patients with PLNM than without PLNM. Logistic regression analysis identified high-level CPE expression as an independent risk factor for PLNM (P = 0.001). Patients with higher CPE expression had shorter overall survival duration than patients with lower CPE expression. Univariate and multivariate Cox-regression analyses suggested that high-level CPE expression is an independent prognostic factor for overall survival in early-stage cervical cancer. CONCLUSIONS: High-level CPE expression was associated with a poor prognosis in early-stage cervical cancer. CPE may serve as a biomarker for predicting PLNM and survival in these patients.

miR-101a and miR-30b contribute to inflammatory cytokine-mediated ß-cell dysfunction.

Inflammatory cytokines have a critical role in the progressive deterioration of pancreatic ß-cell function and development of type 1 diabetes. Prolonged exposure of ß-cells to inflammatory cytokines results in gene expression modifications, leading to loss of ß-cell function. MicroRNAs (miRNAs) are small non-coding RNAs acting as key regulators of gene expression. Here, we demonstrate that miR-101a and miR-30b are key players in cytokine-mediated ß-cell dysfunction. We found that IL-1ß induces an increase in miR-101a and miR-30b in MIN6 cells, and that the two miRNAs participate in ß-cell dysfunction, including decreased insulin content, gene expression, and increased ß-cell death. miR-101a and miR-30b reduce proinsulin expression and insulin content by directly targeting the transcriptional factor Neurod1. In addition, ß-cell apoptosis mediated by miR-101a and miR-30b is associated with diminished expression level of the antiapoptotic protein Bcl2. Moreover, we show that miR-101a causes an impairment in glucose-induced insulin secretion by decreasing the expression of the transcription factor Onecut2. Taken together, our findings suggest that changes in the levels of miR-101a and miR-30b contribute to cytokine-mediated ß-cell dysfunction occurring during the development and progression of type 1 diabetes.

Astrocyte elevated gene-1(AEG-1) induces epithelial-mesenchymal transition in lung cancer through activating Wnt/ß-catenin signaling.

BACKGROUND: Non-small cell lung cancer (NSCLC) is a highly metastatic cancer with limited therapeutic options, so development of novel therapies that target NSCLC is needed. During the early stage of metastasis, the cancer cells undergo an epithelial-mesenchymal transition (EMT), a phase in which Wnt/ß-catenin signaling is known to be involved. Simultaneously, AEG-1 has been demonstrated to activate Wnt-mediated signaling in some malignant tumors. METHODS: Human NSCLC cell lines and xenograft of NSCLC cells in nude mice were used to investigate the effects of AEG-1 on EMT. EMT or Wnt/ß-catenin pathway-related proteins were characterized by western blot, immunofluorescence and immunohistochemistry. RESULTS: In the present study, we demonstrated that astrocyte elevated gene-1(AEG-1) ectopic overexpression promoted EMT, which resulted from the down-regulation of E-cadherin and up-regulation of Vimentin in lung cancer cell lines and clinical lung cancer specimens. Using an orthotopic xenograft-mouse model, we also observed that AEG-1 overexpression in human carcinoma cells led to the development of multiple lymph node metastases and elevated mesenchymal markers such as Vimentin, which is a characteristic of cells in EMT. Furthermore, AEG-1 functioned as a critical protein in the regulation of EMT by directly targeting multiple positive regulators of the Wnt/ß-catenin signaling cascade, including GSK-3ß and CKIδ. Notably, overexpression of AEG-1 in metastatic cancer tissues was closely associated with poor survival of NSCLC patients. CONCLUSIONS: These results reveal the critical role of AEG-1 in EMT and suggest that AEG-1 may be a prognostic biomarker and its targeted inhibition may be utilized as a novel therapy for NSCLC.