SPG21, a potential oncogene targeted by miR-128-3p, amplifies HBx-induced carcinogenesis and chemoresistance via activation of TRPM7-mediated JNK pathway in hepatocellular carcinoma.

PURPOSE: Chronic hepatitis B virus (HBV) infection is the primary risk factor for the malignant progression of hepatocellular carcinoma (HCC). It has been reported that HBV X protein (HBx) possesses oncogenic properties, promoting hepatocarcinogenesis and chemoresistance. However, the detailed molecular mechanisms are not fully understood. Here, we aim to investigate the effects of miR-128-3p/SPG21 axis on HBx-induced hepatocarcinogenesis and chemoresistance. METHODS: The expression of SPG21 in HCC was determined using bioinformatics analysis, quantitative real-time PCR (qRT-PCR), western blotting, and immunohistochemistry (IHC). The roles of SPG21 in HCC were elucidated through a series of in vitro and in vivo experiments, including real-time cellular analysis (RTCA), matrigel invasion assay, and xenograft mouse model. Pharmacologic treatment and flow cytometry were performed to demonstrate the potential mechanism of SPG21 in HCC. RESULTS: SPG21 expression was elevated in HCC tissues compared to adjacent non-tumor tissues (NTs). Moreover, higher SPG21 expression correlated with poor overall survival. Functional assays revealed that SPG21 fostered HCC tumorigenesis and invasion. MiR-128-3p, which targeted SPG21, was downregulated in HCC tissues. Subsequent analyses showed that HBx amplified TRPM7-mediated calcium influx via miR-128-3p/SPG21, thereby activating the c-Jun N-terminal kinase (JNK) pathway. Furthermore, HBx inhibited doxorubicin-induced apoptosis by engaging the JNK pathway through miR-128-3p/SPG21. CONCLUSION: The study suggested that SPG21, targeted by miR-128-3p, might be involved in enhancing HBx-induced carcinogenesis and doxorubicin resistance in HCC via the TRPM7/Ca2+/JNK signaling pathway. This insight suggested that SPG21 could be recognized as a potential oncogene, offering a novel perspective on its role as a prognostic factor and a therapeutic target in the context of HCC.

Surface Segregation Methods toward Molecular Separation Membranes.

As a highly promising approach to solving the issues of energy and environment, membrane technology has gained increasing attention in various fields including water treatment, liquid separations, and gas separations, owing to its high energy efficiency and eco-friendliness. Surface segregation, a phenomenon widely found in nature, exhibits irreplaceable advantages in membrane fabrication since it is an in situ method for synchronous modification of membrane and pore surfaces during the membrane forming process. Meanwhile, combined with the development of synthesis chemistry and nanomaterial, the group has developed surface segregation as a versatile membrane fabrication method using diverse surface segregation agents. In this review, the recent breakthroughs in surface segregation methods and their applications in membrane fabrication are first briefly introduced. Then, the surface segregation phenomena and the classification of surface segregation agents are discussed. As the major part of this review, the authors focus on surface segregation methods including free surface segregation, forced surface segregation, synergistic surface segregation, and reaction-enhanced surface segregation. The strategies for regulating the physical and chemical microenvironments of membrane and pore surfaces through the surface segregation method are emphasized. The representative applications of surface segregation membranes are presented. Finally, the current challenges and future perspectives are highlighted.

Effects of Chlorhexidine-Iodophor Composite Solution on the Viability and Proliferation of Human Skin Fibroblasts Infected by S. aureus - An in Vitro Experiment.

Background: The chlorhexidine-iodophor (CHX-IP) composite solution is a polymer of chlorhexidine and iodophor, applicable to the control of local microbial load and probably toxic to fibroblasts. However, the effect of CHX-IP on the viability and proliferation of human skin fibroblasts infected by Staphylococcus aureus (S. aureus) remains unknown. Objective: The effects of CHX-IP composite solution on the viability and proliferation of human skin fibroblasts infected by S. aureus were investigated in vitro cell culture in this study. Methods: Optimum multiplicity of infection (MOI) was determined to construct the S. aureus-fibroblast co-culture model. Cell Viability Assay was applied to obtain optical density (OD) value and calculate cell viability. 5-ethynyl-2'- deoxyuridine (EdU) assay was used to investigate the effect of CHX-IP on the proliferation of human skin fibroblasts infected by S. aureus. Results: 10:1 was the optimum MOI for the S. aureus-fibroblast co-culture model. The OD value of human skin fibroblasts infected by S. aureus increased in the blank control group, 0.625 mg/ml, 0.3125 mg/ml, 0.15625 mg/ml, and 0.075625 mg/ml groups after four hours. While that of the negative control group, 5 mg/ml, 2.5 mg/ml, and 1.25 mg/ml groups decreased over time. The two-way ANOVA results indicated that the OD value of human skin fibroblasts infected by S. aureus was significantly different among different CHX-IP concentration groups (F = 34.05, P < .001), and the interaction effect between concentration and time was significant (F = 9.442, P < .001). The results of the EdU cell proliferation assay showed that the blank control group, 0.625 mg/ml CHX-IP group, and 0.075625 mg/ml CHX-IP group had an enhanced fibroblasts cell proliferation, while the fibroblasts cell proliferation of the negative control group and 5 mg/ml CHX-IP group was inhibited. Conclusion: The viability and proliferation of human skin fibroblasts infected by S. aureus were inhibited, while specific concentrations of CHX-IP solution can counteract or even reverse the proliferation inhibition effect.

Exosomes as potential diagnosis and treatment for liver cancer.

BACKGROUND: Liver cancer is the fourth most significant cause of cancer-related death. Lack of early diagnosis strategy and a scarcity of efficient therapy constitute the main reasons for its lethality. Exosomes, which contain various bioactive molecules, are characterized by high biocompatibility, low immunogenicity, and high transport efficiency. As a result, exosomes have become a research hotspot and present significant potential for cancer diagnosis biomarkers, biotherapeutics, therapy targets, drug carriers and therapeutic agents. AIM: To explore the potential of exosomes in the diagnosis and treatment of liver cancer. METHODS: We conducted a systematic literature search via PubMed and Web of Science. The following keywords were used: "exosomal biomarkers", "exosomal therapy", "exosomal therapy", and "liver cancer" or "HCC". The duplicate data were deleted by EndNote software. Literature search focused on full-texts and references of each article were carefully checked. One author (Xiao-Cui Wei) screened the literature that met the following inclusion criteria: (1) Detection of exosomes or their contents in clinical samples (body fluid or tissue); or (2) Exosomes served as drug carriers or therapeutic factors. Two authors (Xiao-Cui Wei and Li-Juan Liu) independently reviewed all retained literature and analyzed the information. RESULTS: A total of 1295 studies were identified using the systematic literature search. Of these, 835 duplicate studies were removed. A further 402 irrelevant studies were excluded due to being irrelevant, including other diseases, review articles, the literature containing neither clinical samples nor animal experiments, exosome-independent studies, methods for detecting exosomes, or articles in Chinese. Finally, 58 published papers were retained and analyzed in the study. It showed a list of potential exosomal biomarkers that were upregulated in the blood samples of patients with liver cancer. Those downregulated in exosomes might serve as possible biotherapeutics. Some exosomes derived from cells in vitro were used for cytology or animal experiments to explore the mechanism of these exosome contents in disease. These contents might serve as potential targets for liver cancer. Additionally, we also discussed that exosomes serve as drug carriers or therapeutic factors. CONCLUSION: Exosomes might serve as potential biomarkers or therapeutic biotargets in liver cancer and have the potential to act as drug carriers and self-treatment factors for liver cancer patients.

CPEB1, a novel risk gene in recent-onset schizophrenia, contributes to mitochondrial complex I defect caused by a defective provirus ERVWE1.

BACKGROUND: Schizophrenia afflicts 1% of the world population. Clinical studies suggest that schizophrenia patients may have an imbalance of mitochondrial energy metabolism via inhibition of mitochondrial complex I activity. Moreover, recent studies have shown that ERVWE1 is also a risk factor for schizophrenia. Nevertheless, there is no available literature concerning the relationship between complex I deficits and ERVWE1 in schizophrenia. Identifying risk factors and blood-based biomarkers for schizophrenia may provide new guidelines for early interventions and prevention programs. AIM: To address novel potential risk factors and the underlying mechanisms of mitochondrial complex I deficiency caused by ERVWE1 in schizophrenia. METHODS: Quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay were used to detect differentially expressed risk factors in blood samples. Clinical statistical analyses were performed by median analyses and Mann-Whitney U analyses. Spearman's rank correlation was applied to examine the correlation between different risk factors in blood samples. qPCR, western blot analysis, and luciferase assay were performed to confirm the relationship among ERVWE1, cytoplasmic polyadenylation element-binding protein 1 (CPEB1), NADH dehydrogenase ubiquinone flavoprotein 2 (NDUFV2), and NDUFV2 pseudogene (NDUFV2P1). The complex I enzyme activity microplate assay was carried out to evaluate the complex I activity induced by ERVWE1. RESULTS: Herein, we reported decreasing levels of CPEB1 and NDUFV2 in schizophrenia patients. Further studies showed that ERVWE1 was negatively correlated with CPEB1 and NDUFV2 in schizophrenia. Moreover, NDUFV2P1 was increased and demonstrated a significant positive correlation with ERVWE1 and a negative correlation with NDUFV2 in schizophrenia. In vitro experiments disclosed that ERVWE1 suppressed NDUFV2 expression and promoter activity by increasing NDUFV2P1 level. The luciferase assay revealed that ERVWE1 could enhance the promoter activity of NDUFV2P1. Additionally, ERVWE1 downregulated the expression of CPEB1 by suppressing the promoter activity, and the 400 base pair sequence at the 3' terminus of the promoter was the minimum sequence required. Advanced studies showed that CPEB1 participated in regulating the NDUFV2P1/NDUFV2 axis mediated by ERVWE1. Finally, we found that ERVWE1 inhibited complex I activity in SH-SY5Y cells via the CPEB1/NDUFV2P1/NDUFV2 signaling pathway. CONCLUSION: In conclusion, CPEB1 and NDUFV2 might be novel potential blood-based biomarkers and pathogenic factors in schizophrenia. Our findings also reveal a novel mechanism of ERVWE1 in the etiology of schizophrenia.

Hepatitis B core antigen modulates exosomal miR-135a to target vesicle-associated membrane protein 2 promoting chemoresistance in hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors. The association of hepatitis B virus (HBV) infection with HCC is hitherto documented. Exosomal miRNAs contribute to cancer progression and chemoresistance. HBV X protein has been known to modulate miRNAs that facilitate cell proliferation and the process of hepatocarcinogenesis. However, there has been no report on hepatitis B core antigen (HBc) regulating exosomal miRNAs to induce drug resistance of HCC cells. AIM: To elucidate the mechanism by which HBc promotes Doxorubicin hydrochloride (Dox) resistance in HCC. METHODS: Exosomes were isolated by ultracentrifugation. The morphology and size of exosomes were evaluated by Dynamic Light Scattering (DLS) and transmission electron microscopy (TEM). The miRNAs differentially expressed in HCC were identified using The Cancer Genome Atlas (TCGA) database. The level of miR-135a-5p in patient tissue samples was detected by quantitative polymerase chain reaction. TargetScan and luciferase assay were used to predict and prove the target gene of miR-135a-5p. Finally, we identified the effects of miR-135a-5p on anti-apoptosis and the proliferation of HCC in the presence or absence of Dox using flow cytometry, Cell counting kit 8 (CCK-8) assay and western blot. RESULTS: We found that HBc increased the expression of exosomal miR-135a-5p. Integrated analysis of bioinformatics and patient samples found that miR-135a-5p was increased in HCC tissues in comparison with paracancerous tissues. Bioinformatic analysis and in vitro validation identified vesicle-associated membrane protein 2 (VAMP2) as a novel target gene of miR-135a-5p. Functional assays showed that exosomal miR-135a-5p induced apoptosis protection, cell proliferation, and chemotherapy resistance in HCC. In addition, the rescue experiment demonstrated that VAMP2 reversed apoptosis protection, cell growth, and drug resistance by miR-135a-5p. Finally, HBc promoted HCC anti-apoptosis, proliferation, and drug resistance and prevented Dox-induced apoptosis via the miR-135a-5p/VAMP2 axis. CONCLUSION: These data suggested that HBc upregulated the expression of exosomal miR-135a-5p and promoted anti-apoptosis, cell proliferation, and chemical resistance through miR-135a-5p/VAMP2. Thus, our work indicated an essential role of the miR-135a-5p/VAMP2 regulatory axis in chemotherapy resistance of HCC and a potential molecular therapeutic target for HCC.

Catalytic pyrolysis of poplar sawdust: Excellent hydrocarbon selectivity and activity of hollow zeolites.

Hollow zeolites were investigated for catalytic fast pyrolysis (CFP) of biomass to produce hydrocarbon-rich bio-oil. A series of hollow ZSM-5 catalysts were synthesized via a dissolution-recrystallization strategy. The physicochemical properties of the catalysts were investigated by high-resolution transmission electron microscopy, N2 sorption, X-ray photoelectron spectroscopy, and ammonia temperature-programmed desorption experiments. The hollow zeolite was effective for increasing the hydrocarbon fraction in bio-oil. In particular, hollow HS-ZSM-5(50) afforded the highest hydrocarbon yield (6.8 wt%), which was ~3 times of that achieved with solid ZSM-5(50). The hollowness, acidity, and the presence of secondary wall mesopores in the hollow zeolite were found to affect bio-oil production. The hollow regions stabilized more active biomass intermediates and inhibited their repolymerization to coke, while the interior acid sites continually converted these intermediates to aromatic hydrocarbons. Secondary wall mesopores compromise the hollow space and hinder consecutive catalysis, resulting in phenols as the main product.

Injectable chitosan/ß-glycerophosphate hydrogels with sustained release of BMP-7 and ornidazole in periodontal wound healing of class III furcation defects.

The aim of this study was to determine the effect of bone morphogenetic protein-7 (BMP-7) and ornidazole (ORN) loaded Chitosan/ß-glycerophosphate (CS/ß-GP) thermosensitive hydrogels on periodontal regeneration. CS/ß-GP hydrogels with and without BMP-7 and ORN were compared with respect to physicochemical properties, release kinetics, and antimicrobial activity in vitro, and periodontal regeneration properties in class III furcation defects in beagles via radiography, histology including immunohistochemical staining of osteoblasts and osteoclasts, and histometric analysis. CS/ß-GP hydrogels with and without BMP-7 and ORN had comparable physicochemical properties and gelation kinetics. Release kinetics showed that the hydrogels were capable of stable and sustained release of BMP-7 and ORN. The hydrogels loaded with ORN exhibited obvious antimicrobial activity against P. gingivalis. Histometric analysis quantitatively showed significantly more new bone and cementum, and less connective tissue in defects implanted with BMP-7 loaded hydrogels compared with hydrogels without BMP-7. The number of osteoclasts reduced significantly in the CS/BMP-7/ORN and CS/BMP-7 groups, while the number of osteoblasts increased significantly in these groups. Our findings showed that BMP-7 and ORN conferred additional advantages to the CS/ß-GP hydrogel in periodontal regeneration and suggest potential consideration of this approach for periodontal therapy.

Bone morphogenetic protein 7 promotes odontogenic differentiation of dental pulp stem cells in vitro.

AIMS: in vitro effects of bone morphogenetic protein 7 (BMP-7) on proliferation and differentiation of dental pulp stem cells (DPSCs) have not been investigated, nor has an appropriate dose been established. MAIN METHODS: Human DPSCs obtained from healthy volunteers were cultured with BMP-7 at 25, 50, and 100 ng/ml. Cell viability was measured by Cell Counting Kit-8 assay. Expression profiles of selected odontogenic differentiation-related markers in DPSCs were evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunocytochemistry, and western blot analysis. Mineralization of DPSCs was evaluated by alizarin red staining. The Smad5 signaling pathway was examined by qRT-PCR and western blot analysis. KEY FINDINGS: Diminished cell viability was found in DPSCs induced with 25, 50, and 100 ng/ml of BMP-7 for 7 days, showing a dose-response effect (P-trend = 0.03). DSPP, OCN, DMP-1, and RUNX2 were upregulated by BMP-7 induction after 7 and 14 days, especially at 50 and 100 ng/ml (P < 0.05). Immunocytochemical staining revealed strong expression of DSPP, DMP-1 and ALP in DPSCs induced by BMP-7, whereas null or weak expression in untreated cells. Western blot analysis confirmed over-expression of DSPP in cells induced by BMP-7. Alizarin red staining confirmed formation of mineralized nodules 4 weeks after BMP-7 induction. BMP-7 treated cells showed dose-dependently increased expression of BMPR1A, Smad5, and p-Smad5. SIGNIFICANCE: Our data indicated that BMP-7 at 50 ng/ml and 100 ng/ml was capable to induce DPSCs toward odontogenic differentiation through the Smad5 signaling pathway and not dramatically halt cell proliferation in vitro.

Chitosan composite scaffold combined with bone marrow-derived mesenchymal stem cells for bone regeneration: in vitro and in vivo evaluation.

The study aimed to develop a chitosan (CS)-based scaffold for repairing calvarial bone defects. We fabricated composite scaffolds made of CS and bovine-derived xenograft (BDX), characterized their physicochemical properties including pore size and porosity, absorption, degradation, and compressive strength, compared their efficacy to support in vitro proliferation and differentiation of human jaw bone marrow-derived mesenchymal stem cells (hJBMMSCs), and evaluated their bone regeneration capacity in critical-size rat calvarial defects. The CS/BDX (mass ratio of 40:60) composite scaffold with porosity of 46.23% and pore size of 98.23 µm exhibited significantly enhanced compressive strength than the CS scaffold (59.33 ± 4.29 vs. 18.82 ± 2.49 Kpa). The CS/BDX (40:60) scaffold induced better cell attachment and promoted more osteogenic differentiation of hJBMMSCs than the CS scaffold. The CS/BDX (40:60) scaffold seeded with hJBMMSCs was the most effective in supporting new bone formation, as evidenced by better histomorphometry results, larger new bone area, and more obvious mature lamellar bone formation compared to other groups in rat calvarial defects 8 weeks after implantation. These results suggest that CS/BDX composite scaffold combining with hJBMMSCs has the potential for bone defect regeneration.