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Abstract Background Biopsies obtained from primary oesophageal squamous cell carcinoma (ESCC) guide diagnosis and treatment. However, spatial intra‐tumoral heterogeneity (ITH) influences biopsy‐derived information and patient responsiveness to therapy. Here, we aimed to elucidate the spatial ITH of ESCC and matched lymph node metastasis (LNmet). Methods Primary tumour superficial (PTsup), deep (PTdeep) and LNmet subregions of patients with locally advanced resectable ESCC were evaluated using whole‐exome sequencing (WES), whole‐transcriptome sequencing and spatially resolved digital spatial profiling (DSP). To validate the findings, immunohistochemistry was conducted and a single‐cell transcriptomic dataset was analysed. Results WES revealed 15.72%, 5.02% and 32.00% unique mutations in PTsup, PTdeep and LNmet, respectively. Copy number alterations and phylogenetic trees showed spatial ITH among subregions both within and among patients. Driver mutations had a mixed intra‐tumoral clonal status among subregions. Transcriptome data showed distinct differentially expressed genes among subregions. LNmet exhibited elevated expression of immunomodulatory genes and enriched immune cells, particularly when compared with PTsup (all P

Background: Pyroptosis plays a crucial role in anti-tumor immunity and in formation of the immune microenvironment. However, whether pyroptosis is involved in the progression of clear cell renal cell carcinoma (ccRCC) is still unclear. Personalized treatment of ccRCC requires detailed molecular classification to inform a specific therapy. Methods: Molecular subtyping of ccRCC was performed based on consensus clustering of pyroptosis-related genes. The characteristics of these molecular subtypes were explored at the genome, transcriptome and protein levels. Single-cell RNA sequencing and CIBERSORT analysis were used to analyse the immune microenvironment of ccRCC, while Lasso regression was used to develop a prediction model based on hub genes. Expression of the pyroptosis-related gene GSDMB was also investigated at the tissue and cellular levels. Results: Two molecular subtypes were identified based on the clustering of pyroptosis-related genes. Cluster 1 was associated with activation of classical oncogenic pathways, especially the angiogenesis pathway. Cluster 2 was associated with activation of immune-related pathways and high levels of immunosuppressive cells, exhausted CD8+ T cells, and tumor-associated fibroblast infiltration. Clusters 1 and 2 were thus defined as the angiogenic and inflamed subtypes, respectively. The two subtypes were predictive of the response of ccRCC to anti-angiogenic therapy and immunotherapy, with Cluster 1 patients benefiting from anti-angiogenic therapy and Cluster 2 patients showing better response to anti-PD1 inhibitor therapy. Furthermore, a 9-gene expression signature (HJURP, NUF2, KIF15, MELK, TPX2, PLK1, CDCA3, CTLA4, FOXP3) was identified that could predict outcome and response to immune checkpoint blockade therapy in test cohorts. Finally, GSDMB was found to be involved in the development of renal clear cell carcinoma. Conclusions: These results on pyroptosis-related genes in ccRCC provide a theoretical basis for understanding molecular heterogeneity and for the development of individualized treatment strategies.

RationaleFructose-1,6-bisphosphatase (FBP1) is a tumor suppressor and a key enzyme negatively regulating Warburg effect in cancer. However, regulation of FBP1 protein expression and its exact role in prostate cancer (PCa) is largely unclear. Phosphatase and tensin homolog (PTEN) is one of the most frequently deleted tumor suppressor genes in human PCa. However, the role of PTEN loss in aberrant Warburg effect in cancer remains poorly understood.MethodsExpression of PTEN and FBP1 was analyzed in several PCa cell lines and prostate tumor tissues in mice. Western blot (WB) and RT-PCR approaches were used to examine how PTEN regulates FBP1 expression. Co-immunoprecipitation (co-IP) and in vivo ubiquitination assays were used to define the regulatory mechanisms. A PCa xenograft model was employed to determine the impact of PTEN regulation of FBP1 on PCa growth in vivo.ResultWe demonstrated that in a manner dependent of PI3K/AKT signal pathway PTEN regulated FBP1 expression in various PCa cell lines and tumors in mice. We confirmed that this regulation took place at the protein level and was mediated by SKP2 E3 ubiquitin ligase. Mechanistically, we showed that serine 271 phosphorylation of FBP1 by cyclin-dependent kinases (CDKs) was essential for SKP2-mediated degradation of FBP1 protein induced by PTEN loss. Most importantly, we further showed that loss of PTEN expression enhanced Warburg effect and PCa growth in mice in a manner dependent, at least partially on FBP1 protein degradation.ConclusionsOur results reveal a novel tumor-suppressive feature of PTEN in restraining FBP1 degradation and the Warburg effect. These results also suggest that prohibiting FBP1 protein degradation could be a viable therapeutic strategy for PTEN-deficient PCa.

Vitamin D3 (VD3) participated widely in the nuclear factor-κB (NF-κB)-mediated inflammation, apoptosis, and autophagy through the vitamin D receptor (VDR). However, the molecular mechanisms remain not understood in teleost. The present study investigated the functions of VD3/VDR on intestinal inflammation, autophagy, and apoptosis of turbot in vivo and in vitro. Triple replicates of 30 fish were fed with each of three diets with graded levels of 32.0 (D0), 1012.6 (D1), and 3978.2 (D2) IU/kg VD3. Obvious intestinal enteritis was observed in the D0 group and followed with dysfunction of intestinal mucosal barriers. The intestinal inflammatory response induced by VD3 deficiency was regulated by the NF-κB/inflammasome signalling. The promotion of intestinal apoptosis and suppression of intestinal autophagy were also observed in the D0 group. Similarly, VD3 deficiency in vitro induced more intense inflammation regulated by NF-κB/inflammasome signalling. The mutually exclusive apoptosis and autophagy were also observed in the group without 1,25(OH)2D3in vitro, accompanied by similar changes in apoptosis and autophagy increased apoptosis. The gene expression of VDRs was significantly increased with the increasing VD3 supplementation both in vivo and in vitro. Moreover, VDR knockdown in turbot resulted in intestinal inflammation, and this process relied on the activation of inflammasome mediated by NF-κB signalling. Simultaneously, intestinal apoptosis was promoted, whereas intestinal autophagy was inhibited. In conclusion, VD3 deficiency could induce intestinal inflammation via activation of the NF-κB/inflammasome pathway, intestinal apoptosis, and autophagy formed a mutually exclusive relation in teleost. And VDR is the critical molecule in those processes.

Abstract Background Clear cell renal cell carcinoma (CCRCC) is characterized by a highly metastatic potential. The stromal communication between stem cells and cancer cells critically influences metastatic dissemination of cancer cells. Methods The effect of exosomes isolated from cancer stem cells (CSCs) of CCRCC patients on the progress of epithelial-mesenchymal transition (EMT) and lung metastasis of CCRCC cells were examined. Results CSCs exosomes promoted proliferation of CCRCC cells and accelerated the progress of EMT. Bioactive miR-19b-3p transmitted to cancer cells by CSC exosomes induced EMT via repressing the expression of PTEN. CSCs exosomes derived from CCRCC patients with lung metastasis produced the strongest promoting effect on EMT. Notably, CD103+ CSC exosomes were enriched in tumor cells and in lung as well, highlighting the organotropism conferred by CD103. In addition, CD103+ exosomes were increased in blood samples from CCRCC patients with lung metastasis. Conclusions CSC exosomes transported miR-19b-3p into CCRCC cells and initiated EMT promoting metastasis. CD103+ acted to guide CSC exosomes to target cancer cells and organs, conferring the higher metastatic capacity of CCRCC to lungs, suggesting CD103+ exosomes as a potential metastatic diagnostic biomarker. Graphical abstract ᅟ

ObjectiveTo screen lung adenocarcinoma (LUAC)-specific cell-cycle-related genes (CCRGs) and develop a prognostic signature for patients with LUAC.MethodsThe GSE68465, GSE42127, and GSE30219 data sets were downloaded from the GEO database. Single-sample gene set enrichment analysis was used to calculate the cell cycle enrichment of each sample in GSE68465 to identify CCRGs in LUAC. The differential CCRGs compared with LUAC data from The Cancer Genome Atlas were determined. The genetic data from GSE68465 were divided into an internal training group and a test group at a ratio of 1:1, and GSE42127 and GSE30219 were defined as external test groups. In addition, we combined LASSO (least absolute shrinkage and selection operator) and Cox regression analysis with the clinical information of the internal training group to construct a CCRG risk scoring model. Samples were divided into high- and low-risk groups according to the resulting risk values, and internal and external test sets were used to prove the validity of the signature. A nomogram evaluation model was used to predict prognosis. The CPTAC and HPA databases were chosen to verify the protein expression of CCRGs.ResultsWe identified 10 LUAC-specific CCRGs (PKMYT1, ETF1, ECT2, BUB1B, RECQL4, TFRC, COCH, TUBB2B, PITX1, and CDC6) and constructed a model using the internal training group. Based on this model, LUAC patients were divided into high- and low-risk groups for further validation. Time-dependent receiver operating characteristic and Cox regression analyses suggested that the signature could precisely predict the prognosis of LUAC patients. Results obtained with CPTAC, HPA, and IHC supported significant dysregulation of these CCRGs in LUAC tissues.ConclusionThis prognostic prediction signature based on CCRGs could help to evaluate the prognosis of LUAC patients. The 10 LUAC-specific CCRGs could be used as prognostic markers of LUAC.

Homogeneous functionalization of cellulose with chloroacetyl chloride (CAC), 2-bromoisobutyryl bromide (BrBiB), and 2-chloro-2-phenylacetyl chloride (CPAC) was performed in ionic liquid to evaluate the effect of the molecular structure of the reagents on the reactivity of the cellulosic hydroxyl groups. The results showed that the reaction was very selective for the less hindered C6-OH group, but the substitution of the secondary OH group still occurred, which indicated that the acylation of cellulose was only partly regioselective. The reaction extent and regioselectivity of the cellulosic hydroxyl groups partly depended on the molecular structure of the acylating agents. The reaction rate of the CAC was much faster than the relatively bulky BrBiB and CPAC, but the bulky acylating agents showed a higher C6-OH selectivity. Moreover, the BrBiB was less reactive than the CPAC, although they showed the same regioselectivity for the three hydroxyl groups. The acylation decreased the thermal stability of the cellulose, which decreased further as the bulk of the substituted groups increased.

A dense and uniform Cr coating was fabricated on the zirconium alloys fuel claddings by radio frequency (RF) magnetron sputtering to improve the mechanical and anti-oxidation properties under 1200 °C steam environment. The phase composition and the micro and macro morphologies of the specimens were investigated by X-ray diffraction (XRD), energy dispersive spectroscopy (EDS), scanning electron microscope (SEM) and optical-microtopography (OM) analyses, and the high-temperature oxidation behavior was evaluated at 1200 °C steam environment for 3000 s. In this paper, there exists a positive correlation between thickness and Vickers hardness (HV), and a negative correlation between surface roughness and bonding force. Radial tensile was introduced to investigate the deformation-resistant performance, and the displacement of the Cr-coated specimen was as low as 2.32 mm, which was much lower than the uncoated zircaloy cladding (3.05 mm). Different thicknesses of Cr coatings were deposited to investigate the oxidation degrees of zircaloy cladding under a high-temperature steam environment. The optimal 6 μm Cr-coated zirconium alloys cladding exhibited an excellent anti-oxidation property, and the weight gain was as low as ~4.12 mg/cm2, which was almost one-third of the uncoated specimen.

An amendment to this paper has been published and can be accessed via the original article.

As an effective method of transgenesis, the plasmid of PiggyBac transposon containing GFP (PiggyBac) transposon system has been widely used in various organisms but not yet in mollusks. In this work, piggyBac containing green fluorescent protein (GFP) was transferred into the Pacific oyster (Crassostrea gigas) by sperm-mediated gene transfer with or without electroporation. Fluorescent larvae were then observed and isolated under an inverted fluorescence microscope, and insertion of piggyBac was tested by polymerase chain reaction (PCR) using genomic DNA as template. Oyster larvae with green fluorescence were observed after transgenesis with or without electroporation, but electroporation increased the efficiency of sperm-mediated transgenesis. Subsequently, the recombinant piggyBac plasmid containing gGH (piggyBac-gGH) containing GFP and a growth hormone gene from orange-spotted grouper (gGH) was transferred into oysters using sperm mediation with electroporation, and fluorescent larvae were observed and isolated. The insertion of piggyBac-gGH was tested by PCR and genome walking analysis. PCR analysis indicated that piggyBac-gGH was transferred into oyster larvae; genome walking analysis further showed the detailed location where piggyBac-gGH was inserted in the oyster genome. This is the first time that piggyBac transposon-mediated transgenesis has been applied in mollusks.

Cellulose etherification with 3-chloro-2-hydroxypropyl-trimethylammonium chloride (CHPTAC) was carried out in NaOH/urea aqueous solution, under homogeneous conditions. The substitution degree of the prepared quaternized cellulose, cellulose-CHPTAC, increased with increasing molar ratio of CHPTAC/anhydroglucose unit (AGU), reaction temperature, and reaction time. The structure of the cellulose-CHPTAC was characterized using Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (1H- and 13C-NMR), and thermal gravimetric analysis (TGA). Moreover, a cellulose-CHPTAC sample (DS=0.52) was used to modify ground calcium carbonate (GCC) fillers by the surface wrapping method. The results showed that the filler retention was improved by surface modification of GCC fillers using cellulose-CHPTAC, and the negative impact of filler addition on paper strength was reduced. The quaternized cellulose prepared in NaOH/urea aqueous solution could be considered a new, promising filler modifier.

Background and objective Video-assisted thoracoscopic surgery (VATS) lobectomy is generally accepted for patients with lung cancer. The aim of this study is to explore the feasibility of the single-operation-hole thoracoscopic lobectomy in the treatment of non-small cell lung cancer. Methods To review and analyze the single-operation-hole thoracoscopic lobectomy performed in our hospital for 113 non-small cell lung cancer (NSCLC) cases from October 2010 to October 2013. The incision for observation was 1.5 cm the eighth intercostal at the rear of the midaxillary line and the incision for operation was 2.0 cm-4.0 cm at the fourth or fifth intercostal of the anterior axillary line. The operations were performed through the single-operation-hole. Result The operation processes were smooth for all the patients without any operative mortality occurrence. Only in 5 cases was the operation hole expanded because of the occurrence of massive hemorrhage during the operation; 3 patients with postoperative complications underwent thoracoscopic lobectomy again, including 2 cases of delayed hemorrhage and 1 case of chylothorax. The average surgical duration was (178.24±31.17) min, the average blood loss was (213.56±62.38) mL, and the number of lymph nodes dissected was from 5-22. All diagnose were confirmed by pathology after operation. The average length of stay was (8.17±2.93) d. All cases recovered well during the follow-up of (2-38) months, only 5 cases had recurrence or metastasis. Conclusion The single-operation-hole thomcoscopic lobectomy for lung cancer is safe and feasible, further reducing the trauma, and can be used as a conventional treatment for early- or medium-term NSCLC..

OBJECTIVE: To analyze the clinical results of a partial vertebrectomy with titanium mesh implantation and pedicle screw fixation using a posterior approach to reconstruct the spine in the treatment of thoracolumbar burst fractures. METHOD: From January 2006 to August 2008, 20 patients with severe thoracolumbar fractures were treated.For vertebral bodies associated with one injured intervertebral disk, subtotal vertebrectomy surgery and single-segment fusion were performed. For vertebral bodies with two injured adjacent intervertebral disks, partial vertebrectomy surgery and two-segment fusion were performed. RESULTS: All 20 patients were followed up for 12 to 24 months (average of 18 months). There were no complications such as wound infections, hemopneumothorax or abdominal infections in any of the patients. The neurological status of all of the patients was improved by at least one American Spinal Injury Association grade by the last follow-up. The anterior vertebral body height was an average of 50.77% before surgery, 88.51% after surgery and 87.86% at the last follow up; the sagittal Cobb angle was improved, on average, from 26.15° to 5.39° and was 5.90° at the last follow up. The percentage of spinal stenosis was improved, on average, from 26.07% to 4.93%° and was 6.15% at the last follow up. There were significant differences in the anterior vertebral body height pre- and post-surgery and in the sagittal Cobb angle and the percentage of spinal stenosis (p

Abstract Barkground Circular RNAs (circRNAs) play important regulatory roles in a variety of biological processes in mammals. Multiple birth-traits in goats are affected by several factors, but the expression and function of circRNAs in follicular development of goats are not clear. In this study, we aimed to investigate the possible regulatory mechanisms of circRNA and collected five groups of large follicles (Follicle diameter > 6 mm) and small follicles (1 mm

High and steep sandstone slopes along highway line are at high risk of disasters such as landslides, cracking of support structures, and so on. The monitoring, early warning, and emergency response of such slope disaster face enormous challenges, especially during the rainy season. In this paper, intelligent monitoring, early warning and forecasting system were carried out for the high steep sandstone slope with a transmission line tower at the slope crest along the highway under construction in Guangxi, China. The automatic monitoring data, emergency rescue program and rescue effect were analyzed, and emergency rescue measures for high steep slope protection were taken. The research results show that timely access to disaster warning information can effectively support the analysis of disaster causes and the evaluation of disposal programs. Deep-hole monitoring of deformation characteristics can determine the stable state of slopes, and the tangent angle warning criterion can be used for early warning and prediction of high steep slope landslides. By analyzing the location of the sliding surface and taking timely emergency disposal measures such as layered counterpressure method and micropipes, the landslide activities can be effectively controlled to prevent further acceleration of slope collapse. This study can provide an important reference for the monitoring, early warning, forecasting and emergency rescue of sandstone slopes along highways under construction.

Abstract The progression of muscle development is a pivotal aspect of animal ontogenesis, where miRNA and mRNA exert substantial influence as prominent players. It is important to understand the molecular mechanisms involved in skeletal muscle development to enhance the quality and yield of meat produced by Leizhou goats. We employed RNA sequencing (RNA-SEQ) technology to generate miRNA-mRNA profiles in Leizhou goats, capturing their developmental progression at 0, 3, and 6 months of age. A total of 977 mRNAs and 174 miRNAs were found to be differentially expressed based on our analysis. Metabolic pathways, calcium signaling pathways, and amino acid synthesis and metabolism were found to be significantly enriched among the differentially expressed mRNA in the enrichment analysis. Meanwhile, we found that among these differentially expressed mRNA, some may be related to muscle development, such as MYL10, RYR3, and CSRP3. Additionally,, we identified five muscle-specific miRNAs (miR-127-3p, miR-133a-3p, miR-193b-3p, miR-365-3p, and miR-381) that consistently exhibited high expression levels across all three stages. These miRNAs work with their target genes (FHL3, SESN1, PACSIN3, LMCD1) to regulate muscle development. Taken together, our findings suggest that several miRNAs and mRNAs are involved in regulating muscle development and cell growth in goats. By uncovering the molecular mechanisms involved in muscle growth and development, these findings contribute valuable knowledge that can inform breeding strategies aimed at enhancing meat yield and quality in Leizhou goats.

Vitamin D3 (VD3) participated widely in the nuclear factor-κB (NF-κB)-mediated inflammation, apoptosis, and autophagy through the vitamin D receptor (VDR). However, the molecular mechanisms remain not understood in teleost. The present study investigated the functions of VD3/VDR on intestinal inflammation, autophagy, and apoptosis of turbot in vivo and in vitro. Triple replicates of 30 fish were fed with each of three diets with graded levels of 32.0 (D0), 1012.6 (D1), and 3978.2 (D2) IU/kg VD3. Obvious intestinal enteritis was observed in the D0 group and followed with dysfunction of intestinal mucosal barriers. The intestinal inflammatory response induced by VD3 deficiency was regulated by the NF-κB/inflammasome signalling. The promotion of intestinal apoptosis and suppression of intestinal autophagy were also observed in the D0 group. Similarly, VD3 deficiency in vitro induced more intense inflammation regulated by NF-κB/inflammasome signalling. The mutually exclusive apoptosis and autophagy were also observed in the group without 1,25(OH) 2D3 in vitro, accompanied by similar changes in apoptosis and autophagy increased apoptosis. The gene expression of VDRs was significantly increased with the increasing VD3 supplementation both in vivo and in vitro. Moreover, VDR knockdown in turbot resulted in intestinal inflammation, and this process relied on the activation of inflammasome mediated by NF-κB signalling. Simultaneously, intestinal apoptosis was promoted, whereas intestinal autophagy was inhibited. In conclusion, VD3 deficiency could induce intestinal inflammation via activation of the NF-κB/inflammasome pathway, intestinal apoptosis, and autophagy formed a mutually exclusive relation in teleost. And VDR is the critical molecule in those processes. [ABSTRACT FROM AUTHOR]

This study investigates the effects of varying energy levels in diets on Black Angus steers, focusing on growth performance, muscle composition, rumen microbial community, and their interrelationships. Twenty-seven Black Angus steers, aged approximately 22 months and weighing 520 ± 40 kilograms, were randomly divided into three groups: low-energy (LE), medium-energy (ME), and high-energy (HE). Each group consisted of nine individuals. The steers were fed diets with energy levels of 6.657 MJ/kg (LE), 7.323 MJ/kg (ME), and 7.990 MJ/kg (HE) following a 14-day pre-feeding period, with a subsequent 90-day main experimental phase. After the 90-day feeding period, both the HE and ME groups exhibited significantly higher average daily weight gain (ADG) compared to the LE group (p