Activation of TAZ by XMU-MP-1 inhibits osteoclastogenesis and attenuates ovariectomy-induced cancellous bone loss.

Osteoporosis is a metabolic bone loss disorder usually accompanied by overactivated osteoclast formation and increased bone resorption. Transcriptional co-activator with PDZ-binding motif (TAZ) is an emerging potential target for the treatment of osteoporosis. Our previous research showed that TAZ overexpression inhibited osteoclast formation while TAZ silencing had the opposite effect. In addition, TAZ knockout in mouse osteoclasts induced osteoporosis in animal experiments. XMU-MP-1 (XMU) is a selective MST1/2 inhibitor that can theoretically activate TAZ; however, its effect on osteoporosis remains unknown. In this study, we found that XMU treatment significantly increased TAZ expression in osteoclasts and inhibited osteoclast formation in vitro; however, this inhibitory effect was eliminated after the deletion of TAZ. Furthermore, XMU treatment upregulated TAZ expression in osteoclasts and alleviated ovariectomy (OVX)-induced osteoporosis in bilateral OVX mouse models. These findings suggest that XMU can effectively activate TAZ and that pharmacological activation of TAZ may be a promising option for the treatment of osteoporosis.

Activating transcriptional coactivator with PDZ-binding motif by (R)-PFI-2 attenuates osteoclastogenesis and prevents ovariectomized-induced osteoporosis.

Excessive osteoclast activation is a leading cause of osteoporosis. Therefore, identifying molecular targets and relevant pharmaceuticals that inhibit osteoclastogenesis is of substantial clinical importance. Prior research has indicated that transcriptional coactivator with PDZ-binding motif (TAZ) impedes the process of osteoclastogenesis by engaging the nuclear factor (NF)-κB signaling pathway, thereby suggesting TAZ activation as a potential therapeutic approach to treat osteoporosis. (R)-PFI-2 is a novel selective inhibitor of SETD7 methyltransferase activity, which prevents the nuclear translocation of YAP, a homolog of TAZ. Therefore, we hypothesized that (R)-PFI-2 could be an effective therapeutic agent in the treatment of osteoporosis. To test this hypothesis and explore the underlying mechanism, we first examined the impact of (R)-PFI-2 on osteoclastogenesis in bone marrow macrophages (BMMs) in vitro. (R)-PFI-2 treatment inhibited TAZ phosphorylation induced by NF-κB, thereby enhancing its nuclear localization, protein expression, and activation in BMMs. Moreover, (R)-PFI-2-induced TAZ activation inhibited osteoclast formation in a dose-dependent manner, which involved inhibition of osteoclastogenesis through the TAZ and downstream NF-κB pathways. Furthermore, (R)-PFI-2 inhibited osteoclastogenesis and prevented ovariectomy-induced bone loss in vivo in a mouse model. Overall, our findings suggest that TAZ activation by (R)-PFI-2 inhibits osteoclastogenesis and prevents osteoporosis, indicating an effective strategy for treating osteoclast-induced osteoporosis.

Insight into bacterial role attribution in dissolved organic matter humification during rice straw composting with microbial inoculation.

This study aims to investigate the effect of microbial role distribution in microbial carbon pumps on dissolved organic matter (DOM) humification during rice straw composting with microbial inoculation. Three composting groups were designed, named CK (control), B4 (with Bacillus subtilis, OR058594) and Z1 (with Aspergillus fumigatus, AF202956.1). As a result of inoculation, the composition of microbial communities was changed, so that the microorganisms that promoted DOM humification were concentrated in the responders in the microbial carbon pump. DOM was divided into three components in three composting treatments: C1, C2 and C3. After inoculation with Bacillus subtilis, the C2 component was significantly affected, while after inoculation with Aspergillus fumigatus, the C3 component was significantly affected. The results of physicochemical factors affecting the transformation of DOM fluorescence components indicated that C1, C2 and C3 were related to the abundance of the cellulose-degrading enzyme-encoding gene GH7 in CK and B4 composting. However, the C2 was susceptible to organic matter in Z1 composting. This study explored the distribution of microbial communities from a new perspective, which provided new information for analyzing DOM humification and treating agricultural straws to achieve clean conditions for environmental friendliness.

Dual protective role of velutin against articular cartilage degeneration and subchondral bone loss via the p38 signaling pathway in murine osteoarthritis.

Osteoarthritis (OA) is a common degenerative joint condition associated with inflammation and characterized by progressive degradation of the articular cartilage and subchondral bone loss in the early stages. Inflammation is closely associated with these two major pathophysiological changes in OA. Velutin, a flavonoid family member, reportedly exerts anti-inflammatory effects. However, the therapeutic effects of velutin in OA have not yet been characterized. In this study, we explore the effects of velutin in an OA mouse model. Histological staining and micro-CT revealed that velutin had a protective effect against cartilage degradation and subchondral bone loss in an OA mouse model generated by surgical destabilization of the medial meniscus (DMM). Additionally, velutin rescued IL-1ß-induced inflammation in chondrocytes and inhibited RANKL-induced osteoclast formation and bone resorption in vitro. Mechanistically, the p38 signaling pathway was found to be implicated in the inhibitory effects of velutin. Our study reveals the dual protective effects of velutin against cartilage degradation and subchondral bone loss by inhibiting the p38 signaling pathway, thereby highlighting velutin as an alternative treatment for OA.

Hinokitiol inhibits RANKL-induced osteoclastogenesis in vitro and prevents ovariectomy-induced bone loss in vivo.

Osteoporosis is a metabolic bone-loss disease characterized by abnormally excessive osteoclast formation and bone resorption. Identification of natural medicines that can inhibit osteoclastogenesis, bone resorption, and receptor activator of nuclear factor-κB ligand (RANKL)-induced signaling is necessary for improved treatment of osteoporosis. In this study, hinokitiol, a tropolone-related compound extracted from the heart wood of several cupressaceous plants, was found to inhibit RANKL-induced osteoclast formation and bone resorption in vitro. Hinokitiol inhibited early activation of the ERK, p38, and JNK-MAPK pathways, thereby suppressing the activity and expression of downstream factors (c-Jun, c-Fos, and NFATC1). Consistent with the above in vitro findings, hinokitiol treatment protected against ovariectomy-induced bone loss in vivo. Collectively, our results imply that hinokitiol can potentially serve as an effective agent for treating osteoclast-induced osteoporosis.

MicroRNA-148a-3p alleviates high glucose-induced diabetic retinopathy by targeting TGFB2 and FGF2.

AIMS: Diabetic retinopathy (DR), a common complication of type 1 or type 2 diabetes mellitus, has become the leading cause of blindness among adults in working age. The dysregulation of microRNA has been reported to be strongly related to the initiation or progression of DR. However, neither the biological role nor the molecular mechanism of miR-148a-3p has been researched in DR. This study is designed to investigate the function and mechanism of miR-148a-3p in DR. METHODS: The bioinformatics analysis (Targetscan: https://www.targetscan.org/vert_72/ ) and numerous experiments including real-time quantitative polymerase chain reaction, terminal deoxynucleotidyltransferase dUTP nick end labeling, CCK-8, western blot, vasculogenesis and luciferase reporter assays were used to research the function and mechanism of miR-148a-3p in DR. RESULTS: We constructed DR cell model by treating human retinal microvascular endothelial cells (HRECs) with different concentration gradients of high glucose (HG). Additionally, HG treatment reduced miR-148a-3p level in HRECs. In function, overexpression of miR-148a-3p caused an increase in cell viability and a decrease in cell apoptosis. Besides, miR-148a-3p overexpression led to a damage on blood-retinal barrier (BRB) and suppressed angiogenesis. In mechanism, miR-148a-3p specifically bound to 3' untranslated region of TGFB2 and FGF2. At least, rescue assays demonstrated that the inhibitive influence of miR-148a-3p mimics on BRB injury was offset by overexpression of TGFB2 and the attenuation of angiogenesis resulting from miR-148a-3p mimics was abrogated by overexpression of FGF2 CONCLUSIONS: In a word, we discovered that miR-148a-3p alleviated HG-induced DR by targeting TGFB2 and FGF2. This novel discovery indicated miR-148a-3p as a potential target for DR diagnosis or treatment.

Protective effects of lycium barbarum polysaccharides on blood-retinal barrier via ROCK1 pathway in diabetic rats.

Lycium barbarum polysaccharides (LBP) is commonly known as a traditional Chinese medicine, which has protective effects against diabetic complications in clinic, such as diabetic retinopathy (DR). Previous studies have revealed that Rho/ROCK pathway play an important role in DR development. However, the mechanism between LBP and DR remains unknown. This study aims to explore the clear mechanism of the protective effect of LBP in diabetic retinopathy. In this study, streptozocin (STZ, 45 mg/kg) was administered for diabetic rats modeling. Weight, blood glucose levels and blood lipid were measured to assess the metabolic changes by LBP on diabetic rats. Evans blue (EB) extravasation was determined to assess blood-retinal barrier (BRB) disruption. Hematoxylin and Eosin (HE) staining and immunohistochemistry assay were applied for retina morphology exploration. The membranous disks of retina were examined by transmission electron microscope. Further, high glucose condition was induced in choroidal-retinal endothelial cells (RF/6A). Western blotting was performed for P-Occludin, ROCK1 and P-MLC protein expression. The results indicated that the blood glucose levels, blood lipid and EB infiltration capacity were decreased while the weight was increased in LBP-treated diabetic rats compared with model rats. Moreover, LBP could thicken the overall retina, prevent the disturbance of photoreceptor cell membranous disks and inhibit pathological angiogenesis in diabetes. In addition, the decreased expression of P-Occludin and increased expression of RhoA-associated protein kinase (ROCK) or phosphorylated myosin light chain (P-MLC) were observed in retinal tissue of diabetic rats and high glucose induced by RF/6A cells, which could be rescued by LBP and/or Fasudil. LBP has the protective effects on blood-retinal barrier by regulating the Rho/ROCK signaling pathway in diabetic rats. LBP may be served as a Rho/ROCK inhibitor for the treatment of DR.

miR­133b regulates proliferation and apoptosis in high­glucose­induced human retinal endothelial cells by targeting ras homolog family member A.

The aim of the present study was to investigate the role of microRNA (miR)­133b in high­glucose­induced human retinal endothelial cells (hRECs), particularly regarding its potential targeting of ras homolog family member A (RhoA). To establish the high­glucose­induced diabetic retinopathy (DR) model, hRECs were cultured in high­glucose medium for 1, 2 and 3 days. An Annexin allophycocyanin (APC)/7­aminoactinomycin D (7­AAD) staining assay was performed to measure the apoptosis of hRECs. Next, the cells were transfected with miR­133b inhibitors or mimics, and the cell proliferation and apoptosis were measured by MTT and Annexin­APC/7­AAD staining assays, respectively. In addition, reverse transcription­quantitative polymerase chain reaction (RT­qPCR), western blotting and immunocytochemistry were used to detect the expression levels of RhoA, Rho­associated protein kinase 1 (ROCK1), LIM domain kinase 1 (LIMK), myosin light chain (MLC) and phosphorylated (p)­MLC. It was observed that high­glucose or miR­133b inhibitor treatment attenuated the apoptosis of hRECs, and upregulated the mRNA and protein expression levels of RhoA, ROCK1 and LIMK, as well as the p­MLC protein level, in the hRECs. However, miR­133b overexpression inhibited the cell proliferation, promoted apoptosis, and downregulated the mRNA and protein levels of RhoA, ROCK1 and LIMK, as well as p­MLC protein, in high­glucose­induced hRECs. In conclusion, overexpression of miR­133b inhibited the proliferation and promoted apoptosis in a DR cell model by downregulating RhoA expression.

The involvement of Nrf2 in the protective effects of (-)-Epigallocatechin-3-gallate (EGCG) on NaAsO2-induced hepatotoxicity.

Arsenic exposure produces hepatotoxicity. The common mechanism determining its toxicity is the generation of oxidative stress. Oxidative stress induced by arsenic leads to the activation of nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. (-)-Epigallocatechin-3-gallate (EGCG) possesses a potent antioxidant capacity and exhibits extensive pharmacological activities. This study aims to evaluate effects of EGCG on arsenic-induced hepatotoxicity and activation of Nrf2 pathway. Plasma activities of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase were measured; Histological analyses were conducted to observe morphological changes; Biochemical indexes such as oxidative stress (Catalase (CAT), malonyldialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), reactive oxygen species (ROS)), Nrf2 signaling related genes (Nrf2, Nqo1, and Ho-1) were assessed. The results showed that EGCG inhibited arsenic-induced hepatic pathological damage, liver ROS level and MDA level. Arsenic decreases the antioxidant enzymes SOD, GPX, and CAT activity and the decrease was inhibited by treatment of EGCG. Furthermore, EGCG attenuated the retention of arsenic in liver tissues and improved the expressions of Nrf2 signaling related genes (Nrf2, Nqo1, and Ho-1). These findings provide evidences that EGCG may be useful for reducing hepatotoxicity associated with oxidative stress by the activation of Nrf2 signaling pathway. Our findings suggest a possible mechanism of antioxidant EGCG in preventing hepatotoxicity, which implicate that EGCG may be a potential treatment for arsenicosis therapy.

Influence of artificial luminous environment and TCM intervention on development of myopia rabbits.

OBJECTIVE: To explore the influence of artificial luminous environment and preventive function of traditional Chinese medicine (TCM) intervention based on "Theory of yin-yang clock" on myopia. METHODS: A total of 45 New Zealand young rabbits were randomly divided into 5 groups, 9 for each group. Control group was exposed in natural light. Fluorescent group and full spectrum group were exposed in fluorescent light and full spectrum light, on which basis fluorescent TCM group and full spectrum TCM group were added with "Rizhong Yinyang Formulas", respectively. Optical parameters were measured and the influence of different lights on the serum and retinal dopamine (DA) levels as well as the retinal histopathological tissues was observed. RESULTS: The spectrum of fluorescent light mainly focused at 420-490 nm with the peak value of wavelength near 450 nm, whereas that of full spectrum was wider (400-800 nm) with the peak value near 600 nm. After 4 and 12 weeks, fluorescent group was evidently lower in serum and retinal DA levels (P<0.01), and there was no significant difference among full spectrum group, fluorescent TCM group and full spectrum TCM group (P>0.05). Histopathological observation showed that there was significant difference in pigment epithelium layer, photoreceptor and nerve fiber layer between fluorescent group and control group, but the difference among the test groups was not significant. CONCLUSIONS: Fluorescent light has certain influence on retinal histological construction and visual performance. However, TCM intervention may have some degree of protective function on retina.

Clinical efficacy and prognostic factors of chemoreduction combined with topical treatment for advanced intraocular retinoblastoma.

OBJECTIVE: To explore the clinical efficacy and prognostic factors of chemoreduction combined with topical treatment of advanced intraocular retinoblastoma (RB). MATERIALS AND METHODS: A total of 22 eyes from 17 children with RB were selected for the study and treated with chemoreduction combined with topical cryotherapy, transpupillary thermotherapy (TTT) or episcleral plaque brachytherapy. Clinical and follow-up data were retrospectively analyzed. RESULTS: All children received 2~6 courses of chemoreduction treatment, (4.5±0.8 courses on average); 17 eyes from 13 children were treated by chemoreduction combined with cryotherapy or TTT and 5 eyes from 4 children with chemoreduction combined with 125I episcleral plaque brachytherapy. The eye retention rate was 81.8% (18/22), among which 38.9% (7/18) featured restored or maintained good vision. Postoperative follow-up period was 7 to 34 months, (18.6 ± 5.2 months on average). The recurrence rate was 41.2% (7/17), among which 57.1% (4/7) were controlled by supplementing or appending cryotherapy or TTT treatment during the follow-up period. The tumor basal diameter and thickness were significantly reduced (P<0.05 or P<0.01) after treatment. All children demonstrated different degrees of hair loss, 70.6% (12/17) with different degrees of gastrointestinal reactions, 5.88% (1/17) with neutropenia and 11.8% (2/17) being seriously infected during the chemotherapeutic treatment. Univariate and logistic regression analysis showed that tumor basal diameter before treatment had a significant effect on the prognosis (P<0.01). CONCLUSIONS: Chemoreduction combined with topical therapy can effectively control RB in the short term, and tumor basal diameter before treatment is an independent risk factor for prognosis.

[Genome-wide identification, sequence characteristic and expression analysis of heat shock factors (HSFs) in cucumber].

Heat shock factors (HSFs) are ubiquitous in eukaryotes with diversification in structural feature and biological function in plants. Based on the availability of whole cucumber genome sequences, we characterized the cucumber HSFs gene family which contains at least 21 members. Sequence alignments show that all HSFs possess a specific DNA binding domain (DBD). These HSFs genes are unevenly distributed in the seven cucumber chromosomes except for Csa026480 (located on Scaffold_repeat037858). Phylogenetic analysis shows that HSFs in cucumber could be divided into three families, in which family I included three categories (A, B and C). Phylogenetic tree also reveals nine pairs of orthologous genes and three pairs of paralogous genes, suggesting that HSFs gene family have existed before the separation of cucumber and Arabidopsis thaliana. EST analysis shows that cucumber HSFs may be involved in the development of fruit, female flower and hermaphrodite flower. qRT-PCR analysis demonstrated that these genes exhibit different expression levels in heat stress treatment. These results will provide a foundation for the functions of the HSF gene family.