Expression and functional significance of phosphoenolpyruvate carboxykinase 1 in uveal melanoma.

Uveal melanoma (UVM), an uncommon yet potentially life-threatening ocular cancer, arises from melanocytes in the uveal tract of the eye. The exploration of novel oncotargets for UVM is of paramount importance. In this study, we show that PCK1 (phosphoenolpyruvate carboxykinase 1) expression is upregulated in various UVM tissues as well as in primary UVM cells and immortalized lines. Furthermore, bioinformatics studies reveal that PCK1 overexpression in UVM correlates with advanced disease stages and poor patient survival. Genetic silencing (utilizing viral shRNA) or knockout (via CRISPR/Cas9) of PCK1 significantly curtailed cell viability, proliferation, cell cycle progression, and motility, while provoking apoptosis in primary and immortalized UVM cells. Conversely, ectopic overexpression of PCK1, achieved through a viral construct, bolstered UVM cell proliferation and migration. Gαi3 expression and Akt phosphorylation were reduced following PCK1 silencing or knockout, but increased after PCK1 overexpression in UVM cells. Restoring Akt phosphorylation through a constitutively active mutant Akt1 (S473D) ameliorated the growth inhibition, migration suppression, and apoptosis induced by PCK1 silencing in UVM cells. Additionally, ectopic expression of Gαi3 restored Akt activation and counteracted the anti-UVM cell effects by PCK1 silencing. In vivo, the growth of subcutaneous xenografts of primary human UVM cells was significantly inhibited following intratumoral injection of adeno-associated virus (aav) expressing PCK1 shRNA. PCK1 depletion, Gαi3 downregulation, Akt inhibition, proliferation arrest, and apoptosis were detected in PCK1-silenced UVM xenografts. Collectively, our findings demonstrate that PCK1 promotes UVM cell growth possibly by modulating the Gαi3-Akt signaling pathway.

Effects of Fe-N co-modified biochar on methanogenesis performance, microbial community, and metabolic pathway during anaerobic co-digestion of alternanthera philoxeroides and cow manure.

The performance of anaerobic digestion (AD) is susceptible to disturbances in feedstock degradation, intermediates accumulation, and methanogenic archaea activity. To improve the methanogenesis performance of the AD system, Fe-N co-modified biochar was prepared under different pyrolysis temperatures (300,500, and 700 °C). Meanwhile, pristine and Fe-modified biochar were also derived from alternanthera philoxeroides (AP). The aim was to compare the effects of Fe-N co-modification, Fe modification, and pristine biochar on the methanogenic performance and explicit the responding mechanism of the microbial community in anaerobic co-digestion (coAD) of AP and cow manure (CM). The highest cumulative methane production was obtained with the addition of Fe-N-BC500 (260.38 mL/gVS), which was 42.37 % higher than the control, while the acetic acid, propionic acid, and butyric acid concentration of Fe-N-BC were increased by 147.58 %, 44.25 %, and 194.06 % compared with the control, respectively. The co-modified biochar enhanced the abundance of Chloroflexi and Methanosarcina in the AD system. Metabolic pathway analysis revealed that the increased methane production was related to the formation and metabolism of volatile fatty acids and that Fe-N-BC500 enhanced the biosynthesis of coenzyme A and the cell activity of microorganisms, accelerating the degradation of propionic acid and enhancing the hydrogenotrophic methanogenesis pathway. Overall, Fe-N co-modified biochar was proved to be an effective promoter for accelerated methane production during AD.

Detection of CSTF2 by nano fluorescent probe and its correlation with malignant biological characteristics in liver cancer.

To develop a novel nano DNA fluorescent probe for in situ detection of CSTF2 in liver cancer (LC) and study its correlation with the development of LC, we developed nano-TiO2-DNA fluorescent probe which can bind with CSTF2 in LC samples with high efficiency. The detection process of CSTF2 did not involve the use PCR technology, and the concentration of CSTF2 can be directly observed by fluorescence intensity. This probe exhibited excellent physicochemical properties in ethyl alcohol at -20°C and could directly and selectively permeate into Hep-3B cells. By using CSTF2 Nano-TiO2-DNA probe, we found that the CSTF2 level increased greatly in LC tissue and cells, and high CSTF2 level was closely associated with high levels of tumor markers and poor prognosis in LC patients. After transfection, CSTF2 was overexpressed or silenced in Hep-3B cells, and we find that high CSTF2 level effectively increased the activity and invasion of Hep-3B cells and reduced their apoptosis. Furthermore, high CSTF2 level significantly increased the tumor volume and weight in mice models by activating PI3K/AKT/mTOR signal pathway. Therefore, CSTF2 can serve as an early biomarker of LC and a novel potential target for its treatment.

Low-temperature torrefaction assisted with solid-state KOH/urea pretreatment for accelerated methane production in wheat straw anaerobic digestion.

Low-temperature torrefaction assisted with solid-state KOH/urea applied onto wheat straw was proposed to break down the lignocellulosic material to enhance biomethane production in anaerobic digestion (AD). The optimization of key parameters applying the Box-Behnken design and response surface methodology showed that an addition of 0.1 g/gstraw KOH/urea at 180 °C while torrefying for 30 min was the optimal condition for producing biomethane. Results indicate that co-applying KOH and urea in torrefaction synergistically enhanced the biodegradability of straw by effectively removing lignin and largely retaining cellulose, giving rise to a 41 % increase in the cumulative methane production compared to untreated straw (213 mL/g-volatile solids (VSraw)) from batch AD. Additionally, the nitrogen- and potassium-rich digestates helped to improve soil fertility, thus achieving a zero-waste discharge. This study demonstrated the feasibility of using solid-state KOH/urea assisted low-temperature torrefaction as an effective pretreatment method to promote methane production during AD.

Efficacy and safety of early drain removal following pancreatic resections: a meta-analysis.

BACKGROUND: No consensus was reached with regard to the effect of EDR on postoperative outcomes after pancreatic surgery. The meta-analysis was designed to explore the efficacy and safety of early drain removal (EDR). METHODS: Systematic literature search was performed. Data extraction and correction were performed by three researchers. For dichotomous and continuous outcomes, we calculated the pooled risk difference and mean difference with 95% confidence intervals, respectively. The heterogeneity of included studies was evaluated using Cochran's Q and I2 test. The stratified analyses of pancreaticoduodenectomy (PD) and distal pancreatectomy (DP) were performed. RESULTS: A total of 10 studies including 3 RCTs and 7 non RCTs were included for meta-analysis, among which 1780 patients with EDR and 5613 patients with late drain removal (LDR) were enrolled. The meta-analysis of both all the available studies and studies only with selected low risk patients indicated that EDR group had significantly lower incidences of Grade B/C postoperative pancreatic fistula (POPF) and total complications for both PD and DP. However, no advantages of EDR were observed in the meta-analysis of the 3 RCTs. In addition, EDR was associated with a lower incidence of intra-abdominal infection after PD. While for DP, EDR group had decreased risk of delayed gastric emptying and re-operation, and shorter postoperative in-hospital stay. CONCLUSIONS: The meta-analysis demonstrates that EDR is effective and safe for both PD and DP considering POPF and total complications, especially for patients with low concentration of postoperative drain fluid amylase.

Promoting the overall energy profit through using the liquid hydrolysate during microwave hydrothermal pretreatment of wheat straw as co-substrate for anaerobic digestion.

Liquid hydrolysate (LH) derived from the microwave hydrothermal pretreatment (MHP) of wheat straw (WS) was anaerobically digested together with the solid residual to promote the overall energy profit. Different MHP temperatures (90, 120, 150, 180 °C) and retention times (10, 20, 40 min) were investigated. Increased MHP intensity generated plenty of VFAs (mainly acetate) and phenols in the LH, implying the double-side effect of LH on AD. The highest methane production of 227.92 mL CH4·gVS-1 Raw was obtained with MHP at 120 °C for 10 min, 21.53 % higher than the control. While, MHP at 180 °C for 40 min exhibited 29.02 % lower methane production (113.13 mL CH4·gVS-1 Raw) and 115.86 % longer lag phase (3.13 days) than the control. Butyrate fermentation endowed the treatment groups of 180 °C with resilience from the overload and inhibition. Methanosarcina was largely enriched by the abundant acetate in LH on the early stage of anaerobic digestion (AD), especially when with high MHP intensity. Increased abundance of Methanosaeta and Methanobacterium played a crucial role in maintaining methane production at the middle and later stage. The high number of species and evenness in methanogens community were beneficial for the startup of batch AD. Although negative net energy was obtained, the lower ratio of energy input and output compared with the most researches using the solid residual after MHP as the sole substrate for AD demonstrated the contribution of LH to the overall energy profit.

Safety evaluation of early drain removal following pancreaticoduodenectomy: A single-center retrospective cohort study.

Objectives: The effects of early drain removal (EDR) on postoperative complications after pancreaticoduodenectomy (PD) remains to be investigated. This single-center retrospective cohort study was designed to explore the safety of EDR after PD. Methods: A total of 112 patients undergoing PD with drain fluid amylase (DFA) on postoperative day (POD) 1 and 3 <= 5000 were divided into EDR and late drain removal (LDR). Propensity Score Matching (PSM) was used. We compared postoperative outcomes between two groups and explore the risk factors of total complications using univariate and multiple logistic regression analyses. Results: No statistical differences were found in primary outcomes, including Grade B/C postoperative pancreatic fistula (POPF) (Original cohort: 5.71% vs. 3.90%; P = 1.000; PSM cohort: 3.33% vs. 6.67%; P = 1.000), and total complications (Original cohort: 17.14% vs. 32.47%; P = 0.093; PSM cohort: 13.33% vs. 33.33%; P = 0.067). The EDR was associated with shorter in-hospital stay (Original cohort: 11 days vs. 15 days; P < 0.0001; PSM cohort: 11 days vs. 15 days; P < 0.0001). Conclusions: EDR on POD 3 is safe for patients undergoing PD with low risk of POPF.

Combined effects of liquid digestate recirculation and biochar on methane yield, enzyme activity, and microbial community during semi-continuous anaerobic digestion.

The combined effects of liquid digestate recirculation (LDR) and biochar on methanogenesis and microbial communities were studied in semi-continuous anaerobic reactors fed with wheat straw and swine manure. The tolerated organic loading rate (OLR) was expanded from 5 g- volatile solids (VS)∙L-1∙d-1 in the control to higher than 6 g-VS∙L-1∙d-1 in the LDR. At the OLR of 5.0 g-VS∙L-1∙d-1, average special methane yield in LDR with biochar was 0.234 L∙g-VS-1, which was 5.4 % higher than that of the LDR alone. Moreover, enzyme activity and microbial community analysis indicated that LDR with biochar enhanced the processes of hydrolysis and methanogenesis, and balanced the pathway between hydrogenotrophic and acetoclastic methanogenesis. The co-application of LDR and biochar synergistically enhanced the degradation pathways of substrates and the loading shock resistance of anaerobic digestion system. This study could offer strategies for developing sustainable applications of full and continuous LDR in industrial biogas projects.

Single Cell RNA-Seq Identifies Immune-Related Prognostic Model and Key Signature-SPP1 in Pancreatic Ductal Adenocarcinoma.

There are no reliable biomarkers for early diagnosis or prognosis evaluation in pancreatic ductal adenocarcinoma (PDAC). Multiple scRNA-seq datasets for PDAC were retrieved from online databases and combined with scRNA-seq results from our previous study. The malignant ductal cells were identified through calculating copy number variation (CNV) scores. The robust markers of malignant ductal cells in PDAC were found. Five immune-related signatures, including SPP1, LINC00683, SNHG10, LINC00237, and CASC19, were used to develop a risk score formula to predict the overall survival of PDAC patients. We also constructed an easy-to-use nomogram, combining risk score, N stage, and margin status. The expression level of SPP1 was related to the prognosis and immune regulators. We found that SPP1 was mainly expressed in ductal cells and macrophages in PDAC. In conclusion, we constructed a promising prognostic model based on immune-related signatures for PDAC using scRNA-seq and TCGA_PAAD datasets.

Effects of chlorine disinfectants on the microbial community structure and the performance of anaerobic digestion of swine manure.

The residual chlorine disinfectants (CDs) in swine slurry could negatively impact the anaerobic digestion (AD). The objective of this study was to investigate the effects of CDs on mesophilic and thermophilic AD. The results indicated that CDs exerted inhibition effects on methanogenesis at the initial stage of mesophilic AD, leading to the extension of lag time from 0.62 days for control to 0.85, 1.9, 3.8, and 5.5 days with the increasing CDs concentrations of 50, 100, 200, and 400 mg/L, respectively. Under thermophilic condition, the inhibition effects reduced significantly at the initial stage but a decrease of CMPu at later stage was observed. The microbial analysis revealed that CDs resulted in the enrichment of chlorine-resistant bacteria (Clostridum_sensu_stricto_1) and archaea (Methanosarcina). Addition of activated carbon (AC), zero-valent iron (ZVI) and biochar (BC) was evaluated for alleviating the inhibitions of CDs and proved to be feasible strategies to alleviate the inhibited AD.

The diagnostic role of CT, MRI/MRCP, PET/CT, EUS and DWI in the differentiation of benign and malignant IPMN: A meta-analysis.

PURPOSE: The objective of this study was to assess the diagnostic properties of computed tomography (CT), magnetic resonance imaging (MRI/MRCP) /Magnetic Resonance Cholangiopancreatography (MRCP), positron emission tomography/computed tomography (PET/CT), endoscopic ultrasound (EUS) and diffusion-weighted magnetic resonance imaging (DWI) in distinguishing benign and malignant intraductal papillary mucinous neoplasm (IPMN). MATERIALS AND METHODS: Eligible databases were searched for eligible studies, published through July 2020 on the diagnostic accuracy of these modalities. Diagnostic accuracy parameters, including sensitivity, specificity, diagnostic odds ratio (DOR), and summary receiver operating characteristic curves (SROC) were calculated. Meta-regression was performed to identify the source of heterogeneity. RESULTS: In total, 28 studies were included. Pooled sensitivities for CT, MRI/MRCP, PET/CT, EUS and DWI were 0.7, 0.76, 0.8, 0.6 and 0.72, respectively. Pooled specificities were 0.78, 0.83, 0.9, 0.8 and 0.97. The DORs were 8, 16, 35, 6 and 88. The areas under the curve (AUC) of SROC for CT, MRI/MRCP/MRCP, PET/CT, EUS and DW were 0.8, 0.87, 0.92, 0.79 and 0.82, respectively. CONCLUSION: PET/CT showed the highest AUC and the overall diagnostic accuracy results support the use of MRI/MRCP, PET/CT interchangeably as a first-line examination in the diagnosis of malignant IPMN. With regard to DWI, EUS and CT, each techniques have their advantages and supportive to MRI/MRCP.

The solid-state physicochemical properties and biogas production of the anaerobic digestion of corn straw pretreated by microwave irradiation.

The effect of different temperatures used in microwave pretreatment on enhancing methane production of corn straw was comparatively studied in this paper through the analysis of the physicochemical properties of the pretreated materials and the methane yield during anaerobic digestion. Analytic methods such as scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray diffraction were performed to detect the surface chemistry of the pretreated corn straw. The results indicated that microwave pretreatment could effectively disrupt the lignocellulosic structure to release cellulose, hemicellulose, and related derivatives and make them available for the process of anaerobic digestion. The outcome of the methanogenic assay demonstrated that methane production could be significantly improved by 73.08% concerning the variation of the temperatures in microwave pretreatment. This study provides technical support for pretreatment methods of lignocellulose materials and deems that microwave pretreatment boosts methane yield efficiently during the process of anaerobic digestion of lignocellulosic materials.

MicroRNA-374a promotes pancreatic cancer cell proliferation and epithelial to mesenchymal transition by targeting SRCIN1.

MicroRNAs (miRNAs) play a vital role in the progress of cancer. Whereas the expression and function of miR-374a in pancreatic cancer remain largely unknown. In this study, pancreatic cancer samples and its adjacent normal tissues were obtained from 30 clinical patients with pancreatic cancer. Quantitative real-time PCR (qRT-PCR) was used to measure miR-374a and SRC Kinase Signaling Inhibitor 1 (SRCIN1) expression. Western blotting assay was performed to measure the levels of SRCIN1, E-cadherin, N-cadherin, Vimentin, Zonula occludens-1 (ZO-1) and ß-catenin in PANC-1 cells. Luciferase reporter assay was conducted to confirm the direct targeting of SRCIN1 by miR-374a. Cell proliferation and migration assays were utilized to analyze the role of miR-374a in PANC-1 cells. We found that miR-374a expression was upregulated in pancreatic cancer tissues and cell lines. Over-expression of miR-374a promoted cell proliferation, migration and epithelial-mesenchymal transition (EMT) in pancreatic cancer. While, SRCIN1 expression was downregulated in pancreatic cancer tissues and cells. SRCIN1 was found to be a potential targets of miR-374a by dual-luciferase reporter assay. And SRCIN1 was down-regulated after miR-374a transfection. More than that, over-expression of SRCIN1 inhibited cell proliferation, migration and EMT in pancreatic cancer cell. Therefore, this study revealed that miR-374a promoted cell proliferation, migration and EMT via targeting SRCIN1 in pancreatic cancer.

Input and state estimation for linear systems with a rank-deficient direct feedthrough matrix.

The problem of joint input and state estimation for linear stochastic systems with a rank-deficient direct feedthrough matrix is discussed in this paper. Results from previous studies only solve the state estimation problem; globally optimal estimation of the unknown input is not provided. Based on linear minimum-variance unbiased estimation, a five-step recursive filter with global optimality is proposed to estimate both the unknown input and the state. The relationship between the proposed filter and the existing results is addressed. We show that the unbiased input estimation does not require any new information or additional constraints. Both the state and the unknown input can be estimated under the same unbiasedness condition. Global optimalities of both the state estimator and the unknown input estimator are proven in the minimum-variance unbiased sense.