Metastasis and basement membrane-related signature enhances hepatocellular carcinoma prognosis and diagnosis by integrating single-cell RNA sequencing analysis and immune microenvironment assessment.

BACKGROUND: Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and second leading cause of cancer-related deaths worldwide. The heightened mortality associated with HCC is largely attributed to its propensity for metastasis, which cannot be achieved without remodeling or loss of the basement membrane (BM). Despite advancements in targeted therapies and immunotherapies, resistance and limited efficacy in late-stage HCC underscore the urgent need for better therapeutic options and early diagnostic biomarkers. Our study aimed to address these gaps by investigating and evaluating potential biomarkers to improve survival outcomes and treatment efficacy in patients with HCC. METHOD: In this study, we collected the transcriptome sequencing, clinical, and mutation data of 424 patients with HCC from The Cancer Genome Atlas (TCGA) and 240 from the International Cancer Genome Consortium (ICGC) databases. We then constructed and validated a prognostic model based on metastasis and basement membrane-related genes (MBRGs) using univariate and multivariate Cox regression analyses. Five immune-related algorithms (CIBERSORT, QUANTISEQ, MCP counter, ssGSEA, and TIMER) were then utilized to examine the immune landscape and activity across high- and low-risk groups. We also analyzed Tumor Mutation Burden (TMB) values, Tumor Immune Dysfunction and Exclusion (TIDE) scores, mutation frequency, and immune checkpoint gene expression to evaluate immune treatment sensitivity. We analyzed integrin subunit alpha 3 (ITGA3) expression in HCC by performing single-cell RNA sequencing (scRNA-seq) analysis using the TISCH 2.0 database. Lastly, wound healing and transwell assays were conducted to elucidate the role of ITGA3 in tumor metastasis. RESULTS: Patients with HCC were categorized into high- and low-risk groups based on the median values, with higher risk scores indicating worse overall survival. Five immune-related algorithms revealed that the abundance of immune cells, particularly T cells, was greater in the high-risk group than in the low-risk group. The high-risk group also exhibited a higher TMB value, mutation frequency, and immune checkpoint gene expression and a lower tumor TIDE score, suggesting the potential for better immunotherapy outcomes. Additionally, scRNA-seq analysis revealed higher ITGA3 expression in tumor cells compared with normal hepatocytes. Wound healing scratch and transwell cell migration assays revealed that overexpression of the MBRG ITGA3 enhanced migration of HCC HepG2 cells. CONCLUSION: This study established a direct molecular correlation between metastasis and BM, encompassing clinical features, tumor microenvironment, and immune response, thereby offering valuable insights for predicting clinical outcomes and immunotherapy responses in HCC.

Multifunctional Polypropylene Separator via Cooperative Modification and Its Application in the Lithium-Sulfur Battery.

The continuous shuttling of dissolved polysulfides between the electrodes is the primary cause for the rapid decay of lithium-sulfur batteries. Modulation of the separator-electrolyte interface through separator modification is a promising strategy to inhibit polysulfide shuttling. In this work, we develop a graphene oxide and ferrocene comodified polypropylene separator with multifunctionality at the separator-electrolyte interface. The graphene oxide on the functionalized separator could physically adsorb the polysulfide while the ferrocene component could effectively facilitate the conversion of the adsorbed polysulfide. Due to the combination of these beneficial functionalities, the separator exhibits an excellent battery performance, with a high reversible capacity of 409 mAh g-1 after 500 cycles at 0.2 C. We anticipate that the combinatorial separator functionalization proposed herein is an effective approach for improving the performance of lithium-sulfur batteries.

Effects of zinc and iron on the abundance of Microcystis in Lake Taihu under green light and turbulence conditions.

Trace element is one of the important factors affecting the growth of Microcystis. The effects of zinc (0.4 mg/L) and iron (2 mg/L) on the abundance of Microcystis in Lake Taihu were investigated under continuous turbulence and green light conditions in a microcosm experiment. The study results showed that the abundance of Microcystis in the zinc treatment and the iron treatment group was 8.30% and 214% of that in the control group at the end of the experiment, respectively. The proportion of Cyanobacteria in the total phytoplankton biomass in the control, iron treatment, and zinc treatment group decreased from 99.99% at the beginning of the experiment to 13%, 18%, and 1% at the end of the experiment, respectively. At the end of the microcosm experiment, the phytoplankton community was dominated by Bacillariophyta in the control group, accounting for 63%, but it was dominated by Chlorophyta in the zinc treatment and the iron treatment group, accounting for 89% and 42%, respectively. The study results showed that under green light and turbulence, 0.4 mg/L zinc remarkably decreased the abundance of Microcystis, but 2 mg/L iron effectively increased the number of Microcystis and other algae. This research results provided a new idea for controlling Microcystis blooms.

Continuous hydrodynamic mixing weakens the dominance of Microcystis: evidences from microcosm and lab experiments.

Hydrodynamic mixing is one of the important environment factors in determining phytoplankton community compositions. Here the influences of continuous hydrodynamic mixing on abundance, morphology, and dominance of Microcystis were investigated in microcosm and lab experiments. Our research results showed that Cyanophyta contributed 57.16% to the total biomass in control, but Chlorophyta was the dominant group in continuous hydrodynamic mixing (CHM) group, contributing 76.54% to the total biomass in the microcosm experiment. The average number of Microcystis in control was 1.95 folds in CHM group. However, the mean abundance of Scenedesmus quadricauda and Pseudanabaena limnetica in CHM was 2.47 and 2.97 folds in control. In the lab experiment, the average number of Microcystis flos-aquae in control was 2.97 folds in CHM group. The mean size of M. flos-aquae colony in control (34.65 µm) group were significantly bigger than that in the CHM (26.78 µm) group. This research results demonstrated that continuous hydrodynamic mixing weakened the dominance of Microcystis, but was beneficial for the others algae (S. quadricauda and P. limnetica) and is helpful in understanding the effect of hydrodynamic mixing on Microcystis blooms in freshwater ecosystem.

Cartilage tissue miR-214-3p regulates the TrkB/ShcB pathway paracrine VEGF to promote endothelial cell migration and angiogenesis.

BACKGROUND: This study was designed to explore the mechanisms through which chondrocytes regulated endothelial cell migration and angiogenesis in osteoarthritis (OA). METHODS: The expressions of related genes of OA were detected by Western blot and real-time quantitative PCR. Chondrocytes were co-cultured with endothelial cells, and migration as well as angiogenesis rates, and vascular endothelial growth factor (VEGF) secretion of the cells were detected. The relationship between miRNA and TrkB were analyzed by bioinformatics analysis, RNA immunoprecipitation and dual-luciferase assays. The effects of miRNA on the histopathology of the OA mice were determined. RESULTS: The expressions of NGF, TrkA, TrkB, and ShcB were increased significantly in OA patients. IL-1ß promoted the expressions of TrkA, TrkB, and ShcB in chondrocytes and inhibited the expressions of chondrogenic differentiation markers, but shTrkB partially reversed IL-1ß-mediated chondrogenic differentiation. Overexpression of TrkB promoted cell migration, angiogenesis, and VEGF levels, while silencing ShcB reversed the regulation of TrkB. Moreover, chondrocytes miR-214-3p regulated endothelial cell migration and angiogenesis by targeting TrkB paracrine VEGF to activate PI3K/Akt pathway proteins. In addition, overexpressed miR-214-3p improved collagenase-induced cartilage and synovial damage in OA mice. CONCLUSION: The activation of TrkB/ShcB signaling pathway paracrine VEGF is mediated by miR-214-3p in chondrocytes and it regulates endothelial cell migration and angiogenesis in the development of OA.

LncRNA NEAT1 regulates chondrocyte proliferation and apoptosis via targeting miR-543/PLA2G4A axis.

Osteoarthritis (OA), which is characterized by articular cartilage degeneration, shows a gradually increasing incidence with age. This study explored the molecular mechanism underlying the proliferation and apoptosis of chondrocytes during OA progression. In this study, chondrocytes were isolated from human knee cartilages. The targeted relationship among nuclear enriched abundant transcript 1 (NEAT1), microRNA-543 (miR-543) and PLA2G4A was predicted on TargetScan V7.2 and starBase and validated by performing dual-luciferase reporter assay. High-expressed NEAT1 was detected in OA cartilage and chondrocytes. NEAT1 was negatively correlated with miR-543 and was low-expressed in OA cartilage and PLA2G4A was negatively correlated with miR-543 and was high-expressed in OA cartilage. In OA chondrocytes, the overexpressed NEAT1 inhibited the expressions of p-Akt1 and Bcl-2 and upregulated that of matrix metalloprotease (MMP)-3, MMP-9, MMP-13, interleukin (IL)-6 and IL-8, but such effects of overexpressed NEAT1 were reversed by miR-543 mimic. SiRNA-NEAT1 exerted an opposite effect to NEAT1 overexpression on OA chondrocytes, but this could be reversed by miR-543 inhibitor. The effect of PLA2G4A overexpression was the opposite to miR-543 mimic on OA chondrocytes. In conclusion, NEAT1 could sponge miR-543 to induce PLA2G4A expression, inhibit chondrocyte proliferation and promote apoptosis.

RIPK1 Coordinates Bone Marrow Mesenchymal Stem Cell Survival by Maintaining Mitochondrial Homeostasis via p53.

Survival of mesenchymal stem cells in the bone marrow is essential for bone marrow microenvironment homeostasis, but the molecular mechanisms remain poorly understood. RIPK1 has emerged as a critical molecule of programmed cell death in tissue homeostasis. However, little is known about the regulation of RIPK1 on bone marrow mesenchymal stem cells (MSCs). Here, we have investigated for the first time the role of RIPK1 in bone marrow MSCs. We have found that RIPK1 knockdown suppressed proliferation, differentiation, and migration in bone marrow MSCs. Furthermore, RIPK1 knockdown resulted in the opening of mitochondrial permeability transition pore (mPTP) and mtDNA damage, leading to mitochondrial dysfunction, and consequently induced apoptosis and necroptosis in bone marrow MSCs. Moreover, we identified that the p53-PUMA axis pathway was involved in mitochondrial dysfunction in RIPK1-deficient bone marrow MSCs. Together, our findings highlighted that RIPK1 was indispensable for bone marrow MSC survival.

MicroRNA-613 alleviates IL-1ß-induced injury in chondrogenic CHON-001 cells by targeting fibronectin 1.

BACKGROUND: Osteoarthritis (OA) is an aging-related chronic degenerative joint disease. A number of miRNAs have been found to be involved in the development of OA, but the role of miR-613 in OA remains unclear. Thus, this study aimed to investigate the role of miR-613 during the progression of OA. METHODS: CHON-001 cells were transfected with miR-613 agonist for 48 h, and then exposed to 10 ng/mL IL-1ß for 24 h. Cell viability, cell proliferation and cell apoptosis in CHON-001 cells were assessed by CCK-8, immunofluorescence, and flow cytometry assays, respectively. In addition, the dual luciferase reporter system assay was used to determine the interaction of miR-613 and fibronectin 1 in CHON-001 cells. RESULTS: The level of miR-613 was significantly decreased in IL-1ß-treated CHON-001 cells. Overexpression of miR-613 markedly inhibited IL-1ß-induced apoptosis in CHON-001 cells. In addition, upregulation of miR-613 obviously alleviated IL-1ß-induced inflammatory response and cartilage matrix degradation in CHON-001 cells. Meanwhile, fibronectin 1 was identified as a direct binding target of miR-613 in CHON-001 cells. Overexpression of miR-613 alleviated IL-1ß-induced injury in CHON-001 cells via downregulating the expression of fibronectin 1. Furthermore, overexpression of miR-613 alleviated cartilage degradation, and reduced OARSI scores and subchondral bone thickness in a mouse model of OA. CONCLUSION: Our data indicated that overexpression of miR-613 could inhibit IL-1ß-induced injury in CHON-001 cells via decreasing the level fibronectin 1 in vitro, and alleviate the symptoms of OA in vivo. Therefore, miR-613 might be a potential therapeutic option for the treatment of OA.

Toxicological effects of chlorpyrifos on growth, enzyme activity and chlorophyll a synthesis of freshwater microalgae.

This paper aims to acquire the experimental data on the eco-toxicological effects of agricultural pollutants on the aquatic plants and the data can support the assessment of toxicity on the phytoplankton. The pesticide of Chlorpyrifos used as a good model to investigate its eco-toxicological effect on the different microalgae in freshwater. In order to address the pollutants derived from forestry and agricultural applications, freshwater microalgae were considered as a good sample to investigate the impact of pesticides such as Chlorpyrifos on aquatic life species. Two microalgae of Chlorella pyrenoidosa and Merismopedia sp. were employed to evaluate toxicity of Chlorpyrifos in short time and long time by means of measuring the growth inhibition rate, the redox system and the content of chlorophyll a, respectively. In this study, the results showed that EC50 values ranging from 7.63 to 19.64mg/L, indicating the Chlorpyrifos had a relatively limited to the growth of algae during the period of the acute toxicity experiment. Moreover, when two kinds of algae were exposed to a medium level of Chlorpyrifos, SOD and CAT activities were importantly advanced. Therefore, the growth rate and SOD and CAT activities can be highly recommended for the eco-toxicological assessment. In addition, chlorophyll a also could be used as a targeted parameter for assessing the eco-toxicity of Chlorpyrifos on both Chlorella pyrenoidosa and Merismopedia sp.

Updated Three-Column Concept in surgical treatment for tibial plateau fractures - A prospective cohort study of 287 patients.

PURPOSE: This study introduces an updated Three-Column Concept for the classification and treatment of complex tibial plateau fractures. A combined preoperative assessment of fracture morphology and injury mechanism is utilized to determine surgical approach, implant placement and fixation sequence. The effectiveness of this updated concept is demonstrated through evaluation of both clinical and radiographic outcome measures. PATIENTS AND METHODS: From 2008 to 2012, 355 tibial plateau fractures were treated using the updated Three-Column Concept. Standard radiographic and computed tomography imaging are used to systematically assess and classify fracture patterns as follows: (1) identify column(s) injured and locate associated articular depression or comminution, (2) determine injury mechanism including varus/valgus and flexion/extension forces, and (3) determine surgical approach(es) as well as the location and function of applied fixation. Quality and maintenance of reduction and alignment, fracture healing, complications, and functional outcomes were assessed. RESULTS: 287 treated fractures were followed up for a mean period of 44.5 months (range: 22-96). The mean time to radiographic bony union and full weight-bearing was 13.5 weeks (range: 10-28) and 14.8 weeks (range: 10-26) respectively. The average functional Knee Society Score was 93.0 (range: 80-95). The average range of motion of the affected knees was 1.5-121.5°. No significant difference was found in knee alignment between immediate and 18-month post-operative measurements. Additionally, no significant difference was found in functional scores and range of motion between one, two and three-column fracture groups. Twelve patients suffered superficial infection, one had limited skin necrosis and two had wound dehiscence, that healed with nonoperative management. Intraoperative vascular injury occurred in two patients. Fixation of failure was not observed in any of the fractures treated. CONCLUSION: An updated Three-Column Concept assessing fracture morphology and injury mechanism in tandem can be used to guide surgical treatment of tibial plateau fractures. Limited results demonstrate successful application of biologically friendly fixation constructs while avoiding fixation failure and associated complications of both simple and complex tibial plateau fractures. LEVEL OF EVIDENCE: Level II, prospective cohort study.

A posterior reversed L-shaped approach for the tibial plateau fractures-A prospective study of complications (95 cases).

PURPOSE: This study was aiming to investigate intraoperative and postoperative complications and outcomes of a posterior reversed L-shaped approach (PRLA) in the treatment of the tibial plateau fractures. METHODS: Inclusion criterion was tibial plateau fractures treated through the PRLA either separated or combined with other approaches. Main exclusion criterion was poly-trauma patients. Ninety-five patients with tibial plateau fractures from a prospectively collected database were included and followed in this study. The intraoperative and postoperative complications consisted of vascular and nerve injury, wound infection, dehiscence, haematoma, nonunion, skin necrosis and skin paresthesia. Hospital for Special Surgery Knee Score (HSS) was adopted to evaluate patients' knee function while Short Form 36 Health Survey (SF-36) was used to investigate the patients' general health status. RESULTS: Patients' average age was 46.2 years old (range, 22.0-89.0). The fractures were mainly from high energy injuries involving posterior (and medial) column. 78 of 95 cases were combined with an additional anterolateral approach due to the lateral column involvement. The average follow-up was 52.0 months (range, 12.4-102.6). The total complications rate was 4.2% (4/95). Intraoperative complications occurred in two patients (2.1%). One suffered a popliteal artery injury resulted from an antero-posteriorly drilled K-wire. The patient had a loss of 7° knee extension at one year's follow-up. The other endured an injury of nutrient vessel within the medial head of gastrocnemius. Postoperative skin paresthesia occurred in two patients (2.1%). The other complications associated with the PRLA were not observed. The mean HSS score was 96.1 (range, 80-100). The mean SF-36 score was 94.2 (range, 80-100). CONCLUSIONS: The posterior reversed L-shaped approach allows satisfied visualization of the medial and posterior tibial plateau and has promising clinical results with low complication occurrence. It can be recommended as a routine approach for the treatment of the tibial plateau fractures involving the posterior column.

[N2O emissions and ammonia volatilization from leachate irrigated soils].

The effect of soil properties on N2O emissions and ammonia volatilization after leachate addition was investigated by soil S1 and S2 through incubation experiment. The conversions of soil NH4(+)-N and NO3(-)-N contents after leachate addition were evaluated as well. Soil pH dominantly determines the ammonia volatilization after leachate addition and ammonia volatilization only happens by soil S1 of alkaline nature within the first 5-day during the 10-day incubation. Approximately 3.0% leachate ammonia-N is lost via ammonia volatilization from soil S1. After being added the same amount of leachate ammonia-N, there is an approximately 20-fold gap of N2O emissions between two different soils: soil S1 and S2 (p <0.01). Soil moisture content affects the net generation rate of NO3(-)-N contents substantially and N2O fluxes are thus regulated accordingly. Compared with soils moisture content at water-filled pore space (WFPS) of 46%, N2O fluxes are elevated by 6.5, 1.8, and 2.2 times from the leachate added soil S1 without leachate irrigation (p > 0.05), the leachate added soil S1 (p > 0.05), and the leachate added soil S2 (p <0.05), respectively. During the 10-day incubation, N2O emissions cause 41.1% and 2.3% leachate NH4(+)-N loss from the leachate added soil S1 and S2, respectively. Therefore, avoiding excessively wet conditions (< 70% WFPS) and selecting acidic soil could minimize the potential N2O emissions and ammonia volatilization under leachate irrigation.