Identification of a pyroptosis-immune-related lncRNA signature for prognostic and immune landscape prediction in bladder cancer patients.

PURPOSE: Individualized medicine has become increasingly important in bladder cancer treatment, whereas useful biomarkers for prognostic prediction are still lacking. The current study, therefore, constructed a novel risk model based on pyroptosis- and immune-related long noncoding RNAs (Pyro-Imm lncRNAs) to evaluate the potential prognosis of bladder cancer. METHODS: Corresponding data of bladder cancer patients were downloaded from the Cancer Genome Atlas (TCGA) database. The univariate Cox regression analysis, least absolute shrinkage and selection operator (LASSO) regression analysis, and multivariate Cox regression analysis were employed to establish a predictive signature, which was evaluated by receiver operator characteristic (ROC) analysis and Kaplan-Meier analysis. Furthermore, the immune infiltration, immune checkpoints, and responses to chemotherapeutic drugs were analyzed with this model. RESULTS: Three Pyro-Imm lncRNAs (MAFG-DT, AC024060.1, AC116914.2) were finally identified. Patients in the low-risk group demonstrated a significant survival advantage. The area under the ROC curve (AUC) at 1, 3, and 5 years was 0.694, 0.709, and 0.736 respectively in the entire cohort. KEGG and GO analyses showed that the Wnt pathway plays a crucial role in the high-risk group. The risk score was significantly related to the degree of infiltration of different immune cells, the expression of multiple immune checkpoint genes, and the sensitivity of various chemotherapeutic drugs. CONCLUSION: This novel signature provides a theoretical basis for cancer immunology and chemotherapy, which might help develop individualized therapy.

Analytical performance validation and clinical application of blood gas analyzer on the detection of neonatal bilirubin.

BACKGROUND: It is not yet clear whether the trace blood gas analyzer can be used for biochemical detection of newborns. This study aimed to evaluate the reliability of the method for the detection of bilirubin in infants. METHODS: Based on the Clinical and Laboratory Standards Institute (CLSI) EP15-A2 document, the analytical performance of the blood gas analyzer method for bilirubin detection in neonates was validated. The resulting data of 363 simultaneous bilirubin detection with blood gas analyzer (optical method) and biochemical analyzer (enzymatic method) were reviewed. According to the CLSI EP9-A3 document, the relevance and consistency of the measurement results were evaluated by Pearson correlation analysis, Passing-Bablok regression, and Bland-Altman deviation analysis. RESULTS: The precision and accuracy of the Werfen GEM 4000 blood gas analyzer for the detection of different levels of bilirubin samples adhered to the manufacturer's statement and industry quality standards. The bilirubin detection values of the 2 methods showed a good correlation, and both of them were significantly correlated (P<0.001). Passing-Bablok regression results showed that the regression equation of the bilirubin detection value of the 2 methods is y = -21.00 + 1.17x, with the slope as 1.17 [95% confidence interval (CI): 1.15 to 1.19], and the intercept was -21.00 (95% CI: -23.62 to -18.71), the data of the 2 sets were not consistent in each concentration range. The Bland-Altman plot demonstrated that the bilirubin detection value of 16/363 cases (4.4%) for the 2 methods exceeded the 95% limits of agreement (95% LoA); of which the maximum bias was -30.34 (95% CI: -38.48 to -22.26) and there were 5/76 cases (6.6%) outside the 95% LoA in the >300 µmol/L group. CONCLUSIONS: The method for detecting neonatal total bilirubin by trace blood gas analyzer basically meets the clinical requirements and can be used for the preliminary screening of neonatal jaundice. However, for severe hyperbilirubinemia that requires close monitoring of dynamics, a precise enzymatic quantification is required.

Artesunate Impairs Growth in Cisplatin-Resistant Bladder Cancer Cells by Cell Cycle Arrest, Apoptosis and Autophagy Induction.

Cisplatin, which induces DNA damage, is standard chemotherapy for advanced bladder cancer (BCa). However, efficacy is limited due to resistance development. Since artesunate (ART), a derivative of artemisinin originating from Traditional Chinese Medicine, has been shown to exhibit anti-tumor activity, and to inhibit DNA damage repair, the impact of artesunate on cisplatin-resistant BCa was evaluated. Cisplatin-sensitive (parental) and cisplatin-resistant BCa cells, RT4, RT112, T24, and TCCSup, were treated with ART (1-100 µM). Cell growth, proliferation, and cell cycle phases were investigated, as were apoptosis, necrosis, ferroptosis, autophagy, metabolic activity, and protein expression. Exposure to ART induced a time- and dose-dependent significant inhibition of tumor cell growth and proliferation of parental and cisplatin-resistant BCa cells. This inhibition was accompanied by a G0/G1 phase arrest and modulation of cell cycle regulating proteins. ART induced apoptos is by enhancing DNA damage, especially in the resistant cells. ART did not induce ferroptosis, but led to a disturbance of mitochondrial respiration and ATP generation. This impairment correlated with autophagy accompanied by a decrease in LC3B-I and an increase in LC3B-II. Since ART significantly inhibits proliferative and metabolic aspects of cisplatin-sensitive and cisplatin-resistant BCa cells, it may hold potential in treating advanced and therapy-resistant BCa.

Impact of Nischarin on EMT regulators in breast cancer cell lines.

Nischarin is an integrin-binding protein, which is well known as a novel tumor suppressor. In breast cancer, Nischarin serves a critical role in breast cancer cell migration and invasion. However, the molecular mechanism underlying the role of Nischarin remains unclear. Recent findings have demonstrated that epithelial-mesenchymal transition (EMT) increases the capacity of cell migration and invasion. As a member of the integrin family, it was hypothesized that Nischarin may regulate cellular processes via various signaling pathways associated with the EMT process. The present study detected the mRNA levels of EMT regulators via reverse transcription-quantitative PCR and related protein levels via western blotting in breast cancer cells, following NISCH-overexpression and -knockdown. The results demonstrated that Nischarin inhibits cell proliferation, migration and invasion in breast cancer cells. Furthermore, when the NISCH gene was overexpressed, the relative mRNA level of E-cadherin was increased, while the relative mRNA levels of several transcription factors, such as Snail, ZEB1, N-cadherin, Slug, Twist1 and vimentin, decreased. When NISCH was silenced, these results were reversed. The present results demonstrated that Nischarin suppresses cell migration and invasion via inhibiting the EMT process.

Ethanol production by simultaneous saccharification and cofermentation of pretreated corn stalk.

The recalcitrance of lignocellulosic biomass is a major factor limiting its conversion into biofuels. Therefore, in this study, we pretreated corn stalk with 2% Na2 CO3 and 2% H2 O2 for time 70 min at 130°C using a corn stalk to liquid ratio of 1:10. The fermentation broth from multicopy Saccharomyces cerevisiae strains engineered for lignocellulase synthesis was used to enzymatically hydrolyze the pretreated corn stalk. The highest monosaccharide yield (102 mmol/L) was obtained using 15 IU endocellulase, 10 IU exocellulase, and 15 IU ß-glucosidase per gram of cellulose and 20 IU xylanase per gram of hemicellulose. Subsequently, the high-efficiency ethanol-producing S. cerevisiae strain WXY12, which can produce ethanol from glucose and xylose simultaneously, was added to the fermentation system. The entire process involved ethanol production through simultaneous saccharification and fermentation (SSF). Optimization of the fermentation conditions (yeast powder as a nitrogen source, temperature of 30°C, MgSO4 as a metal ion inducer, rotation speed of 180 rpm, solid-liquid ratio of 1:8, and inoculation amount of 2%) resulted in an ethanol output of 46.87 g/L and a theoretical conversion rate of 27.4%. The results of this study improved corn stalk utilization, reduced hydrolysis costs, and generated high ethanol yields.

The genomic mutation spectrums of breast fibroadenomas in Chinese population by whole exome sequencing analysis.

Fibroadenomas (FAs) are the most common fibroepithelial lesions and the most common benign tumors of the breast in women of reproductive age. Although MED12 mutations, an overwhelming majority of all mutations, and some other gene mutations have been found in FAs, the genomic landscapes of FAs are still not completely clear and the genomic mutation spectrums of FAs in Chinese population remains unknown. Here, by performing whole exome sequencing of 12 FAs and the corresponding normal breast tissues in Chinese Han population, we observed the somatic and germline landscapes of genetic alterations. We identified 16 recurrently mutated genes with 37 nonsynonymous or frameshift somatic mutations and 27 recurrent somatic copy number variants (CNVs). In these mutated genes, MED12 was the most common in FAs, harboring 6 nonsynonymous/frameshift somatic mutations and 1 CNV. In addition, 6 germline mutations of tumor susceptibility genes in 5 FAs were identified and the tumor mutational burden of the 5 FAs was significantly higher than the other 7 FAs without germline mutations. This study provides genomic mutation spectrums of FAs in Chinese population and expand the genetic spectrum of FAs.

The efficacy of a new high intensity focused ultrasound therapy for locally advanced pancreatic cancer.

PURPOSE: To compare the survival benefit and safety of the low power cumulative and traditional high intensity focused ultrasound (HIFU) for locally advanced pancreatic cancer. METHOD: We retrospectively analyzed 38 patients with locally advanced, inoperable, stage III pancreatic patients received HIFU treatment between January 2008 and April 2014 in the Department of Surgery, the Second Affiliated Hospital, Zhejiang University, School of Medicine. 11 of them received the low power cumulative HIFU treatment, while other 27 received the traditional HIFU treatment. The HIFU device used was the FEP-BY02 (Yuande Biomedical Engineering Co. Ltd, Beijing, China). Serum biochemistry and adverse events were assessed before and after treatment. All the patients were followed up until death. The survival rate and adverse events of two groups were compared. RESULTS: In 38 patients, the baseline characteristics including gender, age, Karnofsky performance status (KPS) score, tumor location of two groups were generally well balanced (P > 0.05). The median overall survival (OS) for low power cumulative HIFU group was 10.3 months (95% CI, 6.3-14.3 months), which is significantly longer than traditional HIFU group with 6.0 months (95% CI, 5.2-6.8 months) (P = 0.018). In low power cumulative HIFU group, the 6-month and 12-month survival rates were higher than traditional group, 100% v.s 44.4%, 11.1% v.s 36.4%, respectively. The adverse events in both groups include abdominal pain, fever, C-reactive protein (CRP) elevated. The incidence was lower in low power cumulative HIFU group, however, without statistical significance. CONCLUSION: The low power cumulative HIFU treatment showed a statistical significance in survival benefit with better safety profile compared to the traditional HIFU treatment in patients with locally advanced pancreatic cancer.

Identification of compounds from Paris polyphylla (ChongLou) active against Dactylogyrus intermedius.

The present study was designated to ascertain the anthelmintic activity of the rhizomes of Paris polyphylla and to isolate and characterize the active constituents. The methanol extract from rhizomes of P. polyphylla showed significant anthelmintic activity against Dactylogyrus intermedius with the median effective concentration (EC50) 22.5 mg L(-1). Based on this finding, the methanol extract was fractionated by silica gel column chromatography in a bioassay-guided fractionation yielding 2 bioactive compounds, the structures of these compounds were elucidated as formosanin C and polyphyllin VII. The in vivo tests revealed that formosanin C and polyphyllin VII were significantly effective against D. intermedius with EC50 values of 0.6 and 1.2 mg L(-1), respectively. The acute toxicities (LC50) of formosanin C and polyphyllin VII for grass carp were 2.8 and 2.9 mg L(-1), respectively. The overall results provide important information for the potential application of formosanin C and polyphyllin VII in the therapy of serious infection caused by D. intermedius.

Seroprevalence of rabies virus antibodies in bats from southern China.

Members of the Order Chiroptera are the natural reservoirs of lyssaviruses and play an important role in the transmission of rabies to animals and humans. In this present study, the seroprevalence for rabies virus was determined for bats sampled from four southern provinces on the Chinese mainland. A total of 685 bats of 8 species representing 4 families were collected from 10 sites, and were tested by the indirect fluorescent antibody test using fluorescein isothiocyanate (FITC)-conjugated protein A/G mixture and viral neutralization test. Rabies antibody response was only detected from three bat species (Rousettus leschenaulti, Rhinolophus blythi, and Rhinolophus ferrumequinum). The overall rabies seroconversion rate was only 2.2% (15/685). Of the 15 positive sera, 13 (12 fruit bats and 1 insectivorous bat) were indirect fluorescent antibody test positive, and two insectivorous bats were virus neutralization positive when tested by the modified fluorescent antibody viral neutralization test, albeit extremely low. To our knowledge, this is the first published report describing rabies seroprevalences from Chinese bats. These results suggest that bats may play a role in the ecology of lyssaviruses in China, and further surveillance for the presence of lyssaviruses in bats should be undertaken throughout the country and extended to other species.