The Value of Chemokine and Chemokine Receptors in Diagnosis, Prognosis, and Immunotherapy of Hepatocellular Carcinoma.

Background: Chemokines and chemokine receptors (CCRs) are involved in a variety of anti-tumour and pro-tumour immune processes in vivo, such as angiogenesis, metastasis, proliferation and invasiveness, and influence patient prognosis and response to therapy. Methods: CCRs differentially expressed in HCC and associated with prognosis were extracted from TCGA and GEO databases, and the obtained CCRs were then used to construct signature genes, and the signature gene were selected for expression validation as well as functional experiments to explore the role of CCRs in the treatment and prognosis of HCC. Results: We constructed a prognostic model including five CCRs (CCL20, CCL23, CCR3, CCR10, and CXCR3) and validated the expression of signature genes. The model's risk score is an independent prognostic factor for HCC. We have also developed prognostic model nomograms for clinical use. In addition, we validated that CCR3 expression is associated with poor prognosis in HCC, and the proliferation and migration ability of HCC cells was significantly inhibited after interfering with the expression of CCR3 in MHCC-LM3. We also looked at differences in pathway enrichment, immune infiltration and immune checkpoints. Finally, we found that risk scores were also correlated with drug sensitivity, the high-risk group had a better sensitivity to sorafenib. Conclusion: The CCRs-related gene signature may better assess HCC prognosis and response to immunotherapy and tyrosine kinase inhibitors such as sorafenib in HCC, providing prospective solutions for diagnosis and treatment.

PAFAH1B3 is a KLF9 target gene that promotes proliferation and metastasis in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal human malignancies. Uncontrolled cell proliferation, invasion and migration of pancreatic cancer cells are the fundamental causes of death in PDAC patients. Our previous studies showed that KLF9 inhibits the proliferation, invasion and migration of pancreatic cancer cells. However, the underlying mechanisms are not fully understood. In this study, we found that platelet-activating factor acetylhydrolase IB3 (PAFAH1B3) is highly expressed in pancreatic cancer tissues and cells. In vitro and in vivo studies showed that overexpression of PAFAH1B3 promoted the proliferation and invasion of pancreatic cancer cells, while downregulation of PAFAH1B3 inhibited these processes. We found that KLF9 expression is negatively correlated with PAFAH1B3 expression in pancreatic cancer tissues and cells. Western blotting revealed that KLF9 negatively regulates the expression of PAFAH1B3 in pancreatic cancer tissues and cells. Rescue experiments showed that overexpression of PAFAH1B3 could partially attenuate the suppression of pancreatic cancer cell proliferation, invasion and migration induced by KLF9 overexpression. Finally, chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were carried out, and the results showed that KLF9 directly binds to the promoter of PAFAH1B3 and inhibits its transcriptional activity. In conclusion, our study indicated that KLF9 can inhibit the proliferation, invasion, migration and metastasis of pancreatic cancer cells by inhibiting PAFAH1B3.

Iron-modified biochar inhibiting Cd uptake in rice by Cd co-deposition with Fe oxides in the rice rhizosphere.

Fe-enriched biochar has proven to be effective in reducing Cd uptake in rice plants by enhancing iron plaque formation. However, the effect of Fe on biochar, especially the biochar with high S content, for Cd immobilization in rice rhizosphere was not fully understood. To obtain eco-friendly Fe-loaded biochar at a low cost, garlic straw, bean straw, and rape straw were chosen as the feedstocks for Fe-enhanced biochar production by co-pyrolysis with Fe2O3. The resulting biochars and Fe-loaded biochars were GBC, BBC, BRE, GBC-Fe, BBC-Fe, and BRE-Fe, respectively. XRD and FTIR analyses showed that Fe was successfully loaded onto the biochar. The pristine and Fe-containing biochars were applied at rates of 0% (BC0) and 0.1% in pot experiments. Results suggested that BBC-Fe caused the highest reduction in Cd content of rice grain, and the reductions were 67.9% and 31.4%, compared with BC0 and BBC, respectively. Compared to BBC, BBC-Fe effectively reduced Cd uptake in rice roots by 47.5%. The exchangeable and acid-soluble fraction of Cd (F1-Cd) in soil with BBC-Fe treatment was 37.6% and 63.7% lower than that of BC0 and BBC, respectively. Compared to BC0, soil pH was increased by 0.53 units with BBC-Fe treatment. BBC-Fe significantly increased Fe oxides (free Fe oxides, amorphous Fe oxides, and complex Fe oxides) content in the soil as well. DGT study demonstrated that BBC-Fe could enhance the mobility of sulfate in the rhizosphere, which might be beneficial for Cd fixation in the rhizosphere. Moreover, BBC-Fe increased the relative abundance of Bacteroidota, Firmicutes, and Clostridia, which might be beneficial for Cd immobilization in the rhizosphere. This work highlights the synergistic effect of loaded Fe and biochar on Cd immobilization by enhancing Cd deposited with Fe oxides.

Repetitive temporal interference stimulation improves jump performance but not the postural stability in young healthy males: a randomized controlled trial.

BACKGROUND: Temporal interference (TI) stimulation, an innovative non-invasive brain stimulation technique, has the potential to activate neurons in deep brain regions. The objective of this study was to evaluate the effects of repetitive TI stimulation targeting the lower limb motor control area (i.e., the M1 leg area) on lower limb motor function in healthy individuals, which could provide evidence for further translational application of non-invasive deep brain stimulation. METHODS: In this randomized, double-blinded, parallel-controlled trial, 46 healthy male adults were randomly divided into the TI or sham group. The TI group received 2 mA (peak-to-peak) TI stimulation targeting the M1 leg area with a 20 Hz frequency difference (2 kHz and 2.02 kHz). Stimulation parameters of the sham group were consistent with those of the TI group but the current input lasted only 1 min (30 s ramp-up and ramp-down). Both groups received stimulation twice daily for five consecutive days. The vertical jump test (countermovement jump [CMJ], squat jump [SJ], and continuous jump [CJ]) and Y-balance test were performed before and after the total intervention session. Two-way repeated measures ANOVA (group × time) was performed to evaluate the effects of TI stimulation on lower limb motor function. RESULTS: Forty participants completed all scheduled study visits. Two-way repeated measures ANOVA showed significant group × time interaction effects for CMJ height (F = 8.858, p = 0.005) and SJ height (F = 6.523, p = 0.015). The interaction effect of the average CJ height of the first 15 s was marginally significant (F = 3.550, p = 0.067). However, there was no significant interaction effect on the Y balance (p > 0.05). Further within-group comparisons showed a significant post-intervention increase in the height of the CMJ (p = 0.004), SJ (p = 0.010) and the average CJ height of the first 15 s (p = 0.004) in the TI group. CONCLUSION: Repetitive TI stimulation targeting the lower limb motor control area effectively increased vertical jump height in healthy adult males but had no significant effect on dynamic postural stability.

Synergistically enhancing the selective adsorption of cationic dyes through copper impregnation and amino functionality into iron-based metal-organic frameworks.

Dyes contaminating the sewages have seriously threatened the living beings and their separation from wastewater in terms of potential resource recovery is of high value. Herein, both of metal node doping and ligand group grafting were taken into account to enhance the adsorption selectivity of Fe-MOFs towards cationic dyes. The positive correlation between copper doping amount and selective coefficient (∂MOMB) for methylene blue (MB) over methyl orange (MO) within a certain range was mainly attributed to the increased surface negative charges via partial replacement of Fe(III) with Cu(II). Moreover, the amount of surface negative charges was further increased after amino functionalization and there was a synergism between Cu(II) and -NH2 in selectivity enhancement. As a result, Fe0.6Cu0.4-BDC-NH2 exhibited a 22.5-times increase in ∂MOMB and other cationic dyes including malachite green (MG) and rhodamine B (Rh. B) could also be selectively separated from binary and quaternary mixed dye systems. Moreover, Fe0.6Cu0.4-BDC-NH2 showed many superiorities like a wide pH range of 4.0-8.0, strong anti-interference ability over various inorganic ions, good recyclability, and stability. The adsorption kinetics and isotherm suggested that the MB adsorption process was a homogeneous single-layer chemisorption. Additionally, the thermodynamics manifested that the overall process was exothermic and spontaneous. According to the FT-IR and XPS spectra analysis, the electrostatic interaction and hydrogen bonding were determined as the main driving forces, and π-π interaction also contributed to the adsorption process.

Everolimus inhibits hepatoblastoma by inducing autophagy-dependent ferroptosis.

Everolimus, a known inhibitor of the mammalian target of rapamycin (mTOR), has shown uncertain efficacy in treating hepatoblastoma. This study delves into the potential anti-hepatoblastoma properties of everolimus and its intricate relationship with autophagy and ferroptosis, both in vitro and in vivo. In vivo, tumor tissue from hepatoblastoma patient and human hepatoblastoma cell line HuH-6 were xenografted into nude mice to establish xenograft models for observing the effect of everolimus on tumor growth. In vitro, HuH-6 cells were cultured to evaluate the anti-hepatoblastoma activity of everolimus. Transmission electron microscopy and microtubule-associated proteins 1 light chain 3 (LC3), beclin 1, and p62 protein expressions were employed to investigate autophagy. Additionally, indicators of cell apoptosis, reactive oxygen species (ROS) and proteins associated with ferroptosis were measured to evaluate ferroptosis. The results demonstrate that everolimus treatment effectively induced the formation of autophagosomes in hepatoblastoma cells, upregulated the LC3II/I ratio and beclin 1 expression, and downregulated p62 expression, indicating an enhanced autophagy level both in vitro and in vivo. Furthermore, everolimus treatment induced cell apoptosis, increased ROS level, elevated concentrations of malondialdehyde, 4-hydroxynonenal, and iron content, while reducing the ratio of glutathione/oxidized glutathione, and downregulating the protein expression of glutathione peroxidase 4 and solute carrier family 7 member 11, suggesting its ability to induce ferroptosis in hepatoblastoma cells. Importantly, the induction of ferroptosis by everolimus was significantly reversed in the presence of autophinib, an autophagy inhibitor, indicating the autophagy-dependent of everolimus-induced ferroptosis. Taken together, these findings suggest that everolimus holds promise as an effective anti-hepatoblastoma drug, with its mechanism of action potentially involving the induction of autophagy-dependent ferroptosis in hepatoblastoma cells.

Enhanced Antipediatric Sarcoma Effect of Everolimus with Secukinumab by Targeting IL17A.

In this study, we explored the therapeutic potential of everolimus, an mTOR inhibitor, in a patient-derived xenograft (PDX) of rhabdomyosarcoma, the most prevalent malignant pediatric sarcoma. In addition, rhabdoid tumor cell line A-204 and Ewings sarcoma cell line A-673 were cultured to assess the in vitro effect of everolimus. Furthermore, the cell-derived xenograft (CDX) of A-673 was established and treated with everolimus in vivo. IHC and Western blotting were performed to detect the expressions of pertinent proteins. Results showed that everolimus intervention had limited inhibitory effect on PDX tumor growth compared with cyclophosphamide. Nevertheless, everolimus treatment significantly influenced the phosphorylation levels of S6 kinase beta 1 (S6K1) and eIF4E-binding protein 1 (p-4E-BP1), resulting in the inhibition of angiogenesis in vitro and in vivo. Interestingly, everolimus led to an upregulation in the level of IL17A in sarcoma cells. Notably, when secukinumab, a mAb of IL17A, was combined with everolimus, it synergistically enhanced the inhibitory effect of everolimus on sarcoma cell proliferation in vitro and on the growth of PDX or CDX xenograft tumors in vivo. Importantly, this combination therapy did not affect the mTOR signaling. These results indicate that everolimus exerts an antipediatric sarcoma effect by inhibiting mTOR signal. However, everolimus induces sarcoma cells to produce IL17A, which promotes tumor cell survival and counteracts its antipediatric sarcoma effect. The combination of secukinumab effectively eliminates the effects of IL17A, thereby improving the therapeutic efficacy of everolimus in the context of pediatric sarcomas.

Release of biochar-derived dissolved organic matter and the formation of chlorination disinfection by-products: Effects of pH and chlorine dosage.

Transformation, fragmentation, dissolution, and oxidation of biochar are inevitable in the environment, which will undoubtedly accelerate the release of biochar-derived dissolved organic matter (BDOM) into various water bodies. In addition, biochar may affect disinfection by-products (DBPs) during water treatment and subsequent disinfection. In this study, biochars were derived at three selected pyrolysis temperatures (350 °C, 500 °C, and 650 °C) from rice husk, wheat straw, and shrimp shell, and BDOM was extracted from biochar-derived in artificial seawater and ultrapure water. The TOC analyzer results showed that the concentrations of three BDOM decreased with increasing pyrolysis temperature. The BDOM derived from rice husk biochar and wheat straw biochar in seawater was lower than that in ultrapure water, while that of shrimp shell biochar showed an opposite trend, being released in seawater at nearly twice the extent of that in ultrapure water at 350 °C. Moreover, BDOM showed a catalytic effect on chlorination, and GC analysis showed that the concentrations of its DBPs were affected by pH. The concentration of halogenated acetic acid reached the highest value (54.51 µg/L) in weak acidic environment, while the concentrations of trihalomethane and halogenated acetonitrile had the highest values (23.63 and 47.53 µg/L, respectively) in alkaline conditions. The concentrations of all the three DBPs were lowest under neutral pH conditions. In addition, the volatile halogenated DBPs such as dichloroacetone and trichloroacetone were easily hydrolyzed under alkaline conditions. Therefore, it is something be aware of the release of BDOM and the effects of chlorination DBPs when biochar is used for water treatment or water purification.

Protective strategy for the caudate lobe bile duct during left hemihepatectomy based on imaging data analysis.

Purpose: This study was performed to analyze the rule of confluence of the caudate lobe bile duct (CLD) into the left hepatic duct (LHD) and to discuss the protective strategy during left hemihepatectomy. Methods: MRI of 400 patients and T-tube angiography images of 100 patients were collected, and the imaging rules of the confluence of the CLD into the LHD were summarized. The clinical data of 33 patients who underwent left hemihepatectomy using the protective strategy were analyzed. Results: MRI and T-tube angiography images showed that the length from the confluence point of the CLD into the LHD to the confluence of the left and right hepatic ducts was 1.19 ± 0.40 cm and 1.26 ± 0.39 cm, respectively. The average angle between the longitudinal axis of the 2 bile ducts was 68.27° ± 22.59° and 66.58 ± 22.88°, respectively. Coronal and cross-sectional images showed that inflow from the foot side to the cranial side was noted in 79.8% and 82.0% of patients, respectively, and inflow from the dorsal to the ventral side was observed in 84.5% and 88.0%, respectively. Based on these imaging rules, the safe transection length and plane were summarized, and the CLD was effectively protected in 33 cases of left hemihepatectomy. Conclusion: In left hemihepatectomy, the LHD should be transected at least 1.5 cm away from the confluence of the left and right hepatic ducts, and the plane of transection should be oblique to the dorsal side at an angle of 45° with the LHD, these parameters represent an effective strategy to protect the CLD.

A dihydro-benzo[4,5]imidazo[1,2-c]quinazoline-based probe with aggregation-induced ratiometric emission for the ratiometric fluorescent detection of peroxynitrite in living cells and zebrafish.

As a significant kind of reactive oxygen species (ROS), peroxynitrite (ONOO-) plays an indispensable role in many physiological and pathological processes. This study aimed to synthesize a novel dihydro-benzo[4,5]imidazo[1,2-c]quinazoline-based probe 1 for detecting ONOO-. In 99.5% H2O solution, probe 1 displayed a distinct aggregation-induced ratiometric emission (AIRE), and would selectively respond toward ONOO-via a ratiometric fluorescent signal, along with a short response time (<30 s) and ultra-sensitivity (LOD = 17.6 nM). Moreover, the probe was applied for monitoring the concentration fluctuations of ONOO- in HeLa cells and zebrafish.

Magnetic nanoparticles modified with a copper(i) complex as a novel and efficient reusable catalyst for A3 coupling leading to C-N bond formation.

Propargylamines are an important and valuable family of nitrogen-containing compounds with many applications in the fields of medical, industrial, and chemical processes. One-pot multicomponent A3 coupling reactions of aldehydes, amines, and alkynes in the presence of transition metals as catalysts is an efficient strategy for preparing propargylamines. In this study, we fabricated a novel magnetically reusable copper nanocatalyst [Fe3O4-BIm-Pyrim-CuI] through the immobilization of the copper(i) complex on the surface of the magnetic nanoparticles modified with benzimidazole-pyrimidine ligand and evaluated its catalytic activity in the preparation of propargylamines through one-pot multicomponent A3 coupling reactions of aldehydes, amines, and alkynes. Under this catalytic system, aryl substrates with both electron-donating and electron-withdrawing substituents also gave the desired products in excellent yields under standardized conditions. The Fe3O4-BIm-Pyrim-CuI catalyst was easily separated using an external magnet, and the recovered catalyst was reused in 8 cycles without significant loss of activity.

Brassicasterol inhibits hepatitis B virus-associated hepatocellular carcinoma development via suppression of AKT signaling pathway.

BACKGROUND: Hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) does not respond well to current treatment options like sorafenib, and there is an urgent need for developing therapeutical strategies for HBV + HCC. Brassicasterol has previously shown anti-cancer and anti-viral activities, however, its value against HBV + HCC remains to be explored. METHODS: The inhibitory effect of brassicasterol and sorafenib was evaluated on HBV + HCC cell lines and xenograft mouse model. The cytotoxicity of brassicasterol on normal liver cells were measured by LDH assay. AKT agonist was used to identify the targeted signaling pathway by brassicasterol. RESULTS: Brassicasterol induced HBV + HCC cell death in a both dose-dependent and time-dependent manner, and such inhibition was more potent than sorafenib. Brassicasterol did not show apparent cytotoxicity to normal liver cells. Xenograft mouse model further confirmed the inhibitory effect of brassicasterol on the growth of HBV + HCC. Furthermore, signaling pathway analysis showed that brassicasterol-treated HBV + HCC cells had decreased level of phosphor-AKT expression while the addition of AKT agonist could counteract the inhibitory effect of brassicasterol on HCC, indicating that brassicasterol suppressed AKT pathway to exhibit anti-cancer activity in HBV + HCC cells. In addition, brassicasterol showed similar levels of inhibition on HBV- and HBV + HCC cells. CONCLUSION: Brassicasterol possesses anti-cancer activity against HCC through the downregulation of AKT pathway and such activity is independent of HBV infection.

Pig manure biochar for contaminated soil management: nutrient release, toxic metal immobilization, and Chinese cabbage cultivation.

Pig manure could be an effective fertilizer source for soil, but with high concentrations of xic elements. It has been shown that the pyrolysis method could largely reduce the environmental risk of pig manure. However, the comprehensive analysis of both toxic metals immobilization effect and environmental risk of pig manure biochar applied as a soil amendment is rarely addressed. To address the knowledge gap, this study was carried out with pig manure (PM) and pig manure biochar (PMB). The PM was pyrolyzed at 450 â„ƒ and 700 â„ƒ, the corresponding biochar was abbreviated as PMB450 and PMB700, respectively. The PM and PMB were applied in a pot experiment growing Chinese cabbage (Brassica rape L. ssp. Pekinensis) with clay-loam paddy soil. The application rates of PM were set at 0.5% (S), 2% (L), 4% (M) and 6% (H). With the equivalent mass principle, PMB450 and PMB700 were applied at 0.23% (S), 0.92% (L), 1.84% (M), 2.76% (H), and 0.192% (S), 0.7% (L), 1.4% (M), 2.1% (H), respectively. Parameters of Chinese cabbage biomass and quality, total and available concentrations of toxic metals in soil, and soil chemical properties were systemically measured. The main results of this study showed that compared with PM, PMB700 was more effective than PMB450, which induced the highest reductions of Cu, Zn, Pb, and Cd contents in cabbage by 62.6%, 73.0%, 43.9%, and 74.3%, respectively. Both PM and PMB increased the total contents of metals (Cu, Zn, Pb, and Cd) in soil, and PMB decreased the mobility of Cu, Zn, Pb, and Cd at high application rates (≥2%). Treatment with H-PMB700 reduced CaCl2 extractable Cu, Zn, Pb, and Cd by 70.0%, 71.6%, 23.3%, and 15.9%, respectively. For Cu, Zn, Pb, and Cd fractions with BCR extraction, PMB treatments, especially PMB700, were more effective than PM in decreasing the available fractions (F1 +F2 +F3) at high application rates (≥2%). Overall, pyrolysis with high temperature (e.g., 700 â„ƒ) could significantly stabilize the toxic elements in PM and enhance PM's effect on toxic metals immobilization. The marked effects of PMB700 on toxic metal immobilization and cabbage quality improvement might be attributed to high ash contents and liming effect.

Mechanochemical modified nitrogen-rich biochar derived from shrimp shell: Dominant mechanism in pyridinic-N for aquatic methylene blue removal.

N-doping for the preparation of functional carbon materials is a trending research topic. In this study, N-rich biochar (BC) was prepared by calcining naturally N rich shrimp shells under oxygen-limiting environment, and the calcining temperatures were controlled. BC were activated with 5% hydrochloric acid solutions and then post-modified with ball-milling to obtain a series of novel adsorbents (MBCs). All samples were characterized by SEM, BET, FT-IR, XRD, XPS, TG, and element analysis. Surface area, pore volume, and other surface functional groups were significantly improved after acidizing and ball-milling. The adsorption capacities for MB were MBC350 > MBC500 > MBC650 >BC350 > BC650 > BC500, and the equilibrium adsorption capacities were 575.01 mg/g, 506.52 mg/g, 424.59 mg/g, 113.31 mg/g, 93.53 mg/g and 86.25 mg/g, respectively. The excellent adsorption performance of MBCs for MB was ascribed to Lewis acid-base interaction, π-π interaction, electrostatic interaction and van der Waals, and the quinone group and pyridinic-N on the surface of the MBCs are identified as the major active sites. Taken together, ball-milled shrimp shell biochar is a promising material for cation dye adsorption.

Visible-light-driven iron-based heterogeneous photo-Fenton catalysts for wastewater decontamination: A review of recent advances.

Visible-light-driven heterogeneous photo-Fenton process has emerged as the most promising Fenton-derived technology for wastewater decontamination, owing to its prominent superiorities including the potential utilization of clean energy (solar light), and acceleration of ≡Fe(II)/≡Fe(III) dynamic cycle. As the core constituent, catalysts play a pivotal role in the photocatalytic activation of H2O2 to yield reactive oxidative species (ROS). To date, all types of iron-based heterogeneous photo-Fenton catalysts (Fe-HPFCs) have been extensively reported by the scientific community, and exhibited satisfactory catalytic performance towards pollutants decomposition, sometimes even exceeding the homogeneous counterparts (Fe(II)/H2O2). However, the relevant reviews on Fe-HPFCs, especially from the viewpoint of catalyst-self design are extremely limited. Therefore, this state-of-the-art review focuses on the available Fe-HPFCs in literatures, and gives their classification based on their self-characteristics and modification strategies for the first time. Two classes of representative Fe-HPFCs, conventional inorganic semiconductors of Fe-containing minerals and newly emerging Fe-based metal-organic frameworks (Fe-MOFs) are comprehensively summarized. Moreover, three universal strategies including (i) transition metal (TMs) doping, (ii) construction of heterojunctions with other semiconductors or plasmonic materials, and (iii) combination with supporters were proposed to tackle their inherent defects, viz., inferior light-harvesting capacity, fast recombination of photogenerated carriers, slow mass transfer and low exposure and uneven dispersion of active sites. Lastly, a critical emphasis was also made on the challenges and prospects of Fe-HPFCs in wastewater treatment, providing valuable guidance to researchers for the reasonable construction of high-performance Fe-HPFCs.

A novel AIRE-based fluorescent ratiometric probe with endoplasmic reticulum-targeting ability for detection of hypochlorite and bioimaging.

Hypochlorite (ClO-) plays an important role in the human immune defense system, but high concentrations of ClO- in the endoplasmic reticulum (ER) damage cellular proteins, causing ER stress, cell death, and various diseases. Herein, we developed a simple hydrazone probe (1) featuring aggregation-induced ratiometric emission, which would quickly (within 20 s) and sensitively (detection limit of 15.4 µM) respond to ClO- in an almost pure aqueous solution via a fluorescent ratiometric output. Furthermore, the probe was employed to track the level of ClO- in the ER of HeLa cells and zebrafish.

DLGAP4 acts as an effective prognostic predictor for hepatocellular carcinoma and is closely related to tumour progression.

Disc large associated protein 4 (DLGAP4) plays an important role in neurological diseases, but the role and mechanism of DLGAP4 in hepatocellular carcinoma (HCC) remain unclear. In this study, the prognostic effect of DLGAP4 on HCC patients was investigated by means of bioinformatics. The correlation of DLGAP4 expression with the prognosis of HCC patients was evaluated by TCGA data analysis, and the correlation between DLGAP4 expression and the clinical characteristics of HCC patients was evaluated by the Wilcoxon signed rank test and logistic regression analysis. Kaplan‒Meier and Cox regression methods were used to assess the effect of DLGAP4 expression level on overall survival, and nomograms were used to illustrate the correlation between DLGAP4 gene expression and HCC risk. The genes related to DLGAP4 in HCC were screened, and GO/KEGG enrichment analysis was performed. Furthermore, in vitro and in vivo experiments were conducted to detect the effect of DLGAP4 expression on the proliferation, migration and metastasis of HCC cells. We also examined the effect of DLGAP4 expression on enriched pathway proteins to explore the possible mechanism. The expression levels of DLGAP4 were significantly higher in HCC cell lines and tissue samples than in normal liver cell lines and tissues. The expression of DLGAP4 was significantly associated with clinical characteristics. Survival analysis showed that high expression of DLGAP4 was associated with a poor prognosis in HCC. Multivariate analysis showed that high expression of DLGAP4 was an independent risk factor affecting the overall survival rate in HCC patients. By means of ROC curve analysis and nomograms, we determined the value of DLGAP4 expression in the diagnosis and prognosis evaluation of HCC. GO/KEGG enrichment analysis showed that the PPAR signalling pathway was differentially enriched in patients with high expression of DLGAP4. According to in vitro and in vivo experiments, DLGAP4 knockdown inhibited the proliferation and metastasis of HCC cells and decreased the expression of PPARß/δ protein. In contrast, overexpression of DLGAP4 promoted the proliferation and metastasis of HCC cell, and increased the expression of PPARß/δ protein.In contrast, overexpression of DLGAP4 promoted the proliferation and metastasis of HCC cells and increased the expression of PPARß/δ protein. The results show a close correlation between DLGAP4 expression and clinicopathological features of HCC, and DLGAP4 can be used as a prediction biomarker of HCC.

hsa-miR-34a-5p Ameliorates Hepatic Ischemia/Reperfusion Injury Via Targeting HNF4α.

BACKGROUND: To investigate the relationship between the expression level of hsa-miR-34a-5p and liver injury and to further explore its regulatory signaling pathways Methods: Liver tissue and blood were collected from 60 patients undergoing hepatectomy. We constructed a rat HIRI model and treated it with an intraperitoneal injection of agomir-miR-34a-5p or agomir-normal control (NC) for 7 days after the surgery. The pathological changes of agomir-miR-34a-5p or agomir-normal control (NC) groups were compared. 7702 and AML12 cells were transfected with mimics NC or miR-34a-5p mimics and then treated with H2O2 for 6 hours. Cell apoptosis was detected by flow cytometry, Western blot, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling, respectively. Furthermore, the target genes of miR- 34a-5p were identified by luciferase reporter gene assay and were verified in vitro. RESULTS: The relatively high miR-34a-5p expression group revealed a lower level of alanine aminotransferase and aspartate aminotrans- ferase compared with the relatively low miR-34a-5p expression group. HIRI+agomir-miR-34a-5p rats exhibited significantly higher miR-34a-5p expression, lower serum alanine aminotransferase, aspartate aminotransferase, alleviated hepatic necrosis, reduced hepa- tocyte apoptosis, and decreased expression of apoptosis-related proteins, when compared with HIRI+agomir-NC rats (P < .05). After hydrogen peroxide treatment, alpha mouse liver-12 cell (AML-12) and normal liver cell line LO2 (LO2) cells transfected with miR-34a-5p mimics had significantly lower apoptosis rate compared with miR-34a-5p mimics NC group (P < .05). Hepatocyte nuclear factor 4α was identified as a miR-34a-5p target gene. Hepatocyte nuclear factor 4α expression was significantly downregulated in AML12 and HL-7702 (7702) cells transfected with miR-34a-5p (P < .05). Moreover, AML12 and 7702 cells transfected with miR-34a-5p signifi- cantly showed higher c-Jun N-terminal kinase (JNK), P38, cleavage cas-3, and BCL2 associated X (Bax) protein levels compared with AML12 and 7702 cells transfected with agomir-NC. CONCLUSION: miR-34a-5p possibly protected the liver from I/R injury through downregulating Hepatocyte nuclear factor 4α to inhibit the JNK/P38 signaling pathway.

MiR-27a-3p targets USP46 to inhibit the cell proliferation of hepatocellular carcinoma.

Micro-RNAs are involved in the occurrence and development of hepatocellular carcinoma (HCC) as potential therapeutic targets for HCC. In this study, we found that miR-27a-3p was highly expressed in HCC, which was associated with lower survival rates of HCC patients. In vivo and in vitro functional experiments confirmed that over-expression or knock-down miR-27a-3p could significantly affect the proliferation ability of HCCLM3 and Huh-7, two HCC cell lines. Ubiquitin-specific protease 46 (USP46) was confirmed as the key target gene of miR-27a-3p in HCC via RNA-seq, quantitative polymerase chain reaction, Western blotting, and luciferase report. When knocking down USP46, the proliferation activity of HCC cells was significantly enhanced, while it was significantly inhibited after over-expressing USP4. Above results suggest that the abnormally over-expressed miR-27a-3p in liver promotes the proliferation of cancer cells and accelerates the development of HCC by targeting inhibition the expression of USP46. Targeting miR-27a-3p may be an effective strategy for prevention and treatment of HCC.

A novel indene-chalcone-based fluorescence probe with lysosome-targeting for detection of endogenous carboxylesterases and bioimaging.

An indene-chalcone-based fluorescence probe 1 was synthesized and characterized. Under physiological conditions (containing 5% DMSO), probe 1 showed satisfactory stability with a low background signal and recognized carboxylesterases (CEs) based on the catalytic hydrolysis of ester groups, releasing a significant green fluorescence. Probe 1 presents several features including a short response time (within 20 min), low detection limit (1.3 × 10-4 U/mL) and large stokes shift (over 155 nm). Notably, commercial lysosomal dye co-staining experiments illustrated the lysosomal localization function of 1, with the probe also being used for cell and zebrafish imaging of endogenous CEs.