Are recyclable plastics eco-friendly? Recycled PVC microplastics show higher toxicity than pristine PVC on Vallisneria natans, regardless of Cadmium.

As environmental awareness increases, the use of recyclable plastics has risen. However, it is currently unclear whether recycled microplastics (MPs) pose a lesser or greater environmental risk than pristine MPs. Cadmium (Cd), known for its toxicity to most organisms, can bind with MPs and accumulate in sediments. Few studies have explored the environmental risks posed by the coexistence of recycled MPs and pristine MPs with Cd to rooted macrophytes. We investigated the effects of recycled PVC MPs (R-PVC-MPs) and pristine PVC MPs (PVC-MPs) on Vallisneria natans in the presence and absence of Cd. Results showed that at moderate and high Cd levels, R-PVC-MPs reduced plant Cd enrichment. Despite this, the fresh weight of V. natans exposed to R-PVC-MPs was significantly lower than those exposed to PVC-MPs. Furthermore, R-PVC-MPs had more negative impacts on the physiological traits of V. natans than PVC-MPs, as chlorophyll was significantly reduced across all Cd levels. At high Cd levels, both R-PVC-MPs and PVC-MPs caused significantly high oxidative stress, with no significant differences observed. The PCoA plot showed that different MPs cause noticeable variations within the same Cd concentration. The trait network diagrams illustrated strong interactions among traits, with R-PVC-MPs showing the highest complexity. Lower average degree and decreased edge density indicate that traits of plants with R-PVC-MPs addition are more independent of each other. Our findings suggest that recycled PVC MPs pose a greater environmental risk than pristine PVC MPs, offering reference for assessing the risks of recycled plastics in freshwater ecosystems.

A VMD-BP Model to Predict Laser Welding Keyhole-Induced Pore Defect in Al Butt-Lap Joint.

The detection of keyhole-induced pore positions is a critical procedure for assessing laser welding quality. Considering the detection error due to pore migration and noise interference, this research proposes a regional prediction model based on the time-frequency-domain features of the laser plume. The original plume signal was separated into several signal segments to construct the morphological sequences. To suppress the mode mixing caused by environmental noise, variational modal decomposition (VMD) was utilized to process the signals. The time-frequency features extracted from the decomposed signals were acquired as the input of a backpropagation (BP) neural network to predict the pore locations. To reduce the prediction error caused by pore migration, the effect of the length of the signal segments on the prediction accuracy was investigated. The results show that the optimal signal segment length was 0.4 mm, with an accuracy of 97.77%. The 0.2 mm signal segments failed to eliminate the negative effects of pore migration. The signal segments over 0.4 mm resulted in prediction errors of small and dense pores. This work provides more guidance for optimizing the feature extraction of welding signals to improve the accuracy of welding defect identification.

Preparation of imidazolium ionic liquid functionalized paper membrane for selective extraction of caffeic acid and its structural and functional analogues from Taraxaci Herba.

In the search for pharmaceutically active compounds from natural products, it is crucial and challenging to develop separation or purification methods that target not only structurally similar compounds but also those with specific pharmaceutical functions. The adsorption-based method is widely employed in this field and holds potential for this application, given the diverse range of functional monomers that can be chosen based on structural or functional selectivity. In this work, an imidazolium ionic liquid (IL) modified paper membrane was synthesized via microwave reaction. Caffeic acid (CA), with potential interactions with imidazolium IL and a representative component of phenolic acids in Taraxaci Herba, was chosen as a target compound. After optimization of synthesis and extraction parameters, the resulting extraction membrane could be used to quantitatively analyze CA at ng/ml level, and to extract CA's analogues from the sample matrix. Cheminformatics confirmed the presence of structural and functional similarity among these extracted compounds. This study offers a novel approach to preparing a readily synthesized extraction membrane capable of isolating compounds with structural and functional analogies, as well as developing a membrane solid-phase extraction-based analytical method for natural products.

Appropriate supply of sulfur alleviates lead toxicity and stimulates its accumulation in hyperaccumulator Arabis alpina.

Widespread lead (Pb) contamination of agricultural soils is a global issue stemming from human activities. The remediation of Pb-contaminated soils used for agricultural purposes is critically important to safeguard food crop safety. Despite the modulating effects of sulfur (S) on plant responses to toxic heavy metals, the ecological, physiological, and molecular mechanisms driving such modulation in the Pb hyperaccumulator Arabis alpina L. remain unclear. Here, we investigated the effects of five S concentrations (0, 50, 100, 150, and 200 mg kg-1) on A. alpina grown in Pb-contaminated soil from a lead-zinc mining area. Under S50 (i.e., 50 mg kg-1) and S100 treatments, the Pb concentration in both shoots and roots of A. alpina significantly decreased compared to the control (S0). Specifically, the S50 treatment significantly enhanced Pb accumulation, plant biomass, and plant height, indicating that low S applications facilitate Pb accumulation from the soil and alleviate Pb toxicity. Additionally, S50, S100, and S150 treatments significantly improved photosynthetic rate, stomatal conductance, and intercellular CO2 concentration in A. alpina. Transcriptomic analysis showed that S50 and S100 treatments increased the expression of the LHCA, LHCB, psa, and psb genes, which had a significant impact on photosynthetic efficiency. S50 and S100 boosted glutathione (GSH) levels in A. alpina roots, and the increased expression of GST gene enhanced tolerance to environmental stress. In summary, these results suggest that an appropriate supply of S (S50 and S100) not only alleviates Pb toxicity by enhancing plant biomass, height, photosynthetic features, and sulfur metabolites but also stimulates Pb accumulation in the hyperaccumulator A. alpina. Our study elucidated the specific concentrations of sulfur that optimally enhance both Pb accumulation and stress tolerance in the hyperaccumulator A. alpina, providing novel insights into the practical application of sulfur in phytoremediation strategies and advancing our understanding of the underlying molecular mechanisms.

Preparation of imidazole-modified paper membrane for selective extraction of gallic acid and its structural and functional analogues from Pomegranate Peel.

In the search for pharmaceutically active compounds from natural products, it is crucial and challenging to develop separation methods that target not only structurally similar compounds but also a class of compounds with desired pharmaceutical functions. To achieve both structure-oriented and function-oriented selectivity, the choice of functional monomers with broad interactions or even biomimetic roles towards targeted compounds is essential. In this work, an imidazole (IM)-functionalized paper membrane was synthesized to realize selectivity. The IM was selected based on its capability to provide multiple interactions, participation in several bioprocesses, and experimental verification of adsorption performance. Using gallic acid as a representative component of Pomegranate Peel, the preparation conditions and extraction parameters were systematically investigated. The optimal membrane solid-phase extraction (MSPE) method allowed for enrichment of gallic acid from the complex matrix of Pomegranate Peel, enabling facile quantitative analysis with a limit of detection (LOD) of 0.1 ng mL-1. Furthermore, with the aid of cheminformatics, the extracted compounds were found to be similar in both their structures and pharmaceutical functions. This work offers a novel approach to preparing a readily synthesized extraction membrane capable of isolating compounds with similar structures and pharmaceutical effects, and provides an MSPE-based analytical method for natural products.

lncRNA Helf promotes hepatic inflammation and fibrosis by interacting with PTBP1 to facilitate PIK3R5 mRNA stabilization.

BACKGROUND: Hepatic fibrosis is a common consequence of chronic liver diseases without approved antifibrotic therapies. Long noncoding RNAs (lncRNAs) play an important role in various pathophysiological processes. However, the functions of certain lncRNAs involved in mediating the antifibrotic role remain largely unclear. METHODS: The RNA level of lnc-High Expressed in Liver Fibrosis (Helf) was detected in both mouse and human fibrotic livers. Furthermore, lnc-Helf-silenced mice were treated with carbon tetrachloride (CCl4) or bile duct ligation (BDL) to investigate the function of lnc-Helf in liver fibrosis. RESULTS: We found that lnc-Helf has significantly higher expression in human and mouse fibrotic livers as well as M1 polarized hepatic macrophages (HMs) and activated hepatic stellate cells (HSCs). In vivo studies showed that silencing lnc-Helf by AAV8 vector alleviates CCl4- and BDL-induced hepatic inflammation and fibrosis. Furthermore, in vitro experiments revealed that lnc-Helf promotes HSCs activation and proliferation, as well as HMs M1 polarization and proliferation in the absence or presence of cytokine stimulation. Mechanistically, our data illustrated that lnc-Helf interacts with RNA binding protein PTBP1 to promote its interaction with PIK3R5 mRNA, resulting in increased stability and activating the AKT pathway, thus promoting HSCs and HMs activation and proliferation, which augments hepatic inflammation and fibrosis. CONCLUSION: Our results unveil a lnc-Helf/PTBP1/PIK3R5/AKT feedforward, amplifying signaling that exacerbates the process of hepatic inflammation and fibrosis, thus providing a possible therapeutic strategy for hepatic fibrosis.

A Recombinant Oncolytic Pseudorabies Virus Expressing Interleukin-18, Interferon-Gamma and PH20 Genes Promotes Systemic Antitumor Immunity.

Pseudorabies virus (PRV) is considered to be a promising oncolytic virus that has potential as a cancer gene therapy drug. In this study, PRV-DCD-1-70 was used as a vector to carry exogenous genes IL-18, IFN-γ and PH20 to construct novel recombinant PRV, rPRV-PH20 and rPRV-IL-18-γ-PH20, and their tumorolytic effects were evaluated in vitro and in vivo. Our study showed that recombinant PRV lysed all four tumor cell lines, Pan02, EMT-6, CT26 and H446, and rPRV-IL-18-γ-PH20 showed the best tumor lysis effect. Further studies in mice bearing Pan02 tumors showed that recombinant PRV, especially rPRV-IL-18-γ-PH20, were able to inhibit tumor growth. Moreover, an immunohistochemical analysis indicated that the recombinant PRV effectively increased the infiltration of CD4+T and CD8+T cells and enhanced the anti-tumor immune response of the organism in vivo. Overall, PRV carrying PH20 and IL-18-γ exogenous genes demonstrated anti-tumor effects, providing a foundation for the further development and application of PRV as a novel tumor oncolytic virus vector.

Study on Corrosion Resistance of Stainless-Steel Welded Joints with SnSb8Cu4 and SnZn9.

The use of soldering based on metallurgical bonding, as opposed to conventional rubber sealing, is capable of achieving the firm sealing of stainless-steel subway car bodies, though the corrosion resistance of such joints has rarely been investigated. In this study, two typical solders were selected and applied to the soldering of stainless steel, and their properties were investigated. As indicated by the experimental results, the two types of solder exhibited favorable wetting and spreading properties on stainless-steel plates, and successfully achieved sealing connections between the stainless-steel sheets. In comparison with the Sn-Zn9 solder, the Sn-Sb8-Cu4 solder exhibited lower solidus-liquidus, such that it can be more suitably applied to low-temperature sealing brazing. The sealing strength of the two solders reached over 35 MPa, notably higher than that of the sealant currently used (the sealing strength is lower than 10 MPa). In comparison with the Sn-Sb8-Cu4 solder, the Sn-Zn9 solder exhibited a higher corrosion tendency and a higher degree of corrosion during the corrosion process.

Buccal plate preservation with immediate post-extraction implant placement and provisionalization in anterior maxillary tooth: Preliminary results of a new technique using Teruplug collagen.

BACKGROUND: Bone resorption and remodelling are inevitable results of dental extraction and begin immediately after the extraction procedure. The buccal plate is especially predisposed to these phenomena, and if affected, may result in an increased risk of facial soft-tissue recession and other adverse clinical effects that may decrease the predictability of implant placement or impair the final aesthetic result. PERPOSE: The application of Teruplug collagen to prevent buccal plate resorption technique is a new technique aimed at maintaining or improving the appearance of the soft and hard tissues after dental extraction procedures. METHODS: All patients underwent teeth extraction and buccal plate preservation followed by immediate implant placement and provisionalisation using Teruplug collagen. The distance from the external surface of the labial bone to the buccal surface of the implant was measured immediately after placement 6 months and 12 months using computed tomography(CT) images. The aesthetic outcome of 35 implant supported dentures was evaluated by the pink aesthetic score (PAS). Patient aesthetic satisfaction was investigated by a visual analogue scale (VAS). RESULTS: In a four-wall intact socket, this approach is aimed at optimising the ability of the Teruplug collagen to improve regeneration and maintain or improve labial/buccal contours without interfering with the natural healing capability of the alveolus after extraction and implant placement. During the different observation period, there were no major biologic or prosthodontic complications as determined by a clinical examination at each follow-up visit. CONCLUSIONS: Buccal plate preservation as described may help to maintain or improve the appearance and contours of the ridge after tooth extraction, laying the groundwork for optimal functional and aesthetic replacement of the missing tooth with an implant-supported prosthesis.

Differential effects of dexmedetomidine on Gram-positive and Gram-negative bacterial killing and phagocytosis.

Dexmedetomidine is a commonly used sedative in perioperative and intensive care settings with purported immunomodulatory properties. Since its effects on immune functions against infections have not been extensively studied, we tested the effects of dexmedetomidine on Gram-positive [Staphylococcus aureus and Enterococcus faecalis] and Gram-negative bacteria [Escherichia coli], and on effector functions of human monocytes THP-1 cells against them. We evaluated phagocytosis, reactive oxygen species (ROS) formation, and CD11b activation, and performed RNA sequencing analyses. Our study revealed that dexmedetomidine improved Gram-positive but mitigated Gram-negative bacterial phagocytosis and killing in THP-1 cells. The attenuation of Toll-like receptor 4 (TLR4) signaling by dexmedetomidine was previously reported. Thus, we tested TLR4 inhibitor TAK242. Similar to dexmedetomidine, TAK242 reduced E. coli phagocytosis but enhanced CD11b activation. The reduced TLR4 response potentially increases CD11b activation and ROS generation and subsequently enhances Gram-positive bacterial killing. Conversely, dexmedetomidine may inhibit the TLR4-signaling pathway and mitigate the alternative phagocytosis pathway induced by TLR4 activation through LPS-mediated Gram-negative bacteria, resulting in worsened bacterial loads. We also examined another α2 adrenergic agonist, xylazine. Because xylazine did not affect bacterial clearance, we proposed that dexmedetomidine may have an off-target effect on bacterial killing process, potentially involving crosstalk between CD11b and TLR4. Despite its potential to attenuate inflammation, we provide a novel insight into potential risks of dexmedetomidine use during Gram-negative infections, highlighting the differential effect of dexmedetomidine on Gram-positive and Gram-negative bacteria.

LncRNA Airn maintains LSEC differentiation to alleviate liver fibrosis via the KLF2-eNOS-sGC pathway.

BACKGROUND: Long noncoding RNAs (lncRNAs) have emerged as important regulators in a variety of human diseases. The dysregulation of liver sinusoidal endothelial cell (LSEC) phenotype is a critical early event in the fibrotic process. However, the biological function of lncRNAs in LSEC still remains unclear. METHODS: The expression level of lncRNA Airn was evaluated in both human fibrotic livers and serums, as well as mouse fibrotic livers. Gain- and loss-of-function experiments were performed to detect the effect of Airn on LSEC differentiation and hepatic stellate cell (HSC) activation in liver fibrosis. Furthermore, RIP, RNA pull-down-immunoblotting, and ChIP experiments were performed to explore the underlying mechanisms of Airn. RESULTS: We have identified Airn was significantly upregulated in liver tissues and LSEC of carbon tetrachloride (CCl4)-induced liver fibrosis mouse model. Moreover, the expression of AIRN in fibrotic human liver tissues and serums was remarkably increased compared with healthy controls. In vivo studies showed that Airn deficiency aggravated CCl4- and bile duct ligation (BDL)-induced liver fibrosis, while Airn over-expression by AAV8 alleviated CCl4-induced liver fibrosis. Furthermore, we revealed that Airn maintained LSEC differentiation in vivo and in vitro. Additionally, Airn inhibited HSC activation indirectly by regulating LSEC differentiation and promoted hepatocyte (HC) proliferation by increasing paracrine secretion of Wnt2a and HGF from LSEC. Mechanistically, Airn interacted with EZH2 to maintain LSEC differentiation through KLF2-eNOS-sGC pathway, thereby maintaining HSC quiescence and promoting HC proliferation. CONCLUSIONS: Our work identified that Airn is beneficial to liver fibrosis by maintaining LSEC differentiation and might be a serum biomarker for liver fibrogenesis.

Association of X-linked TLR-7 gene polymorphism with the risk of knee osteoarthritis: a case-control study.

Knee osteoarthritis (OA) is the most prevalent type of OA, and Toll-like receptor 7 (TLR7) may lead to the pathogenesis of OA. Recently, X-linked TLR7 polymorphism has been confirmed to be associated with arthritis. However, there is a lack of studies on TLR7 gene polymorphism associated with knee OA susceptibility. The current study aimed to determine whether TLR7 gene polymorphism is associated with the risk of knee OA. Genotyping of two polymorphic sites (rs3853839 and rs179010) in the TLR7 gene was performed in 252 OA patients, and 265 healthy controls using the SNaPshot sequencing technique. Data were analyzed statistically by Chi-square tests and logistic regression. Rs3853839-C allele showed frequencies of 28% and 27% in the healthy control and female knee OA groups, respectively. The differences were not statistically significant (P > 0.05). The rs3853839-CG genotype frequency was significantly lower in the female knee OA group as compared to the healthy control group (OR 0.60; 95%CI 0.36-0.99; P = 0.044). In the male hemizygote population, the rs3853839-CC showed significantly lower frequencies in the male knee OA group as compared to the healthy control group (OR 0.35; 95%CI 0.17-0.71; P = 0.0025). Regarding rs179010, there were no differences in the genotype distribution and allele frequencies between OA patients and healthy subjects under any models (P > 0.05). Stratified analysis showed that the frequency of the rs3853839-CG genotypes was lower in high Kellgren-Lawrence grades (KLG) (OR 0.48; 95%CI 0.21-1.08; P = 0.066), and significantly lower in OA patients with effusion synovitis (OR 0.38; 95%CI 0.17-0.88; P = 0.013). TLR7 rs3853839 polymorphism may play a role in the susceptibility of knee OA in the Chinese Han Population and may be associated with OA severity and the risk of effusion synovitis in Knee OA.

Comparative Transcriptomics Analysis of Roots and Leaves under Cd Stress in Calotropis gigantea L.

Calotropis gigantea is often found in mining areas with heavy metal pollution. However, little is known about the physiological and molecular response mechanism of C. gigantea to Cd stress. In the present study, Cd tolerance characteristic of C. gigantea and the potential mechanisms were explored. Seed germination test results showed that C. gigantea had a certain Cd tolerance capacity. Biochemical and transcriptomic analysis indicated that the roots and leaves of C. gigantea had different responses to early Cd stress. A total of 176 and 1618 DEGs were identified in the roots and leaves of C. gigantea treated with Cd compared to the control samples, respectively. Results indicated that oxidative stress was mainly initiated in the roots of C. gigantea, whereas the leaves activated several Cd detoxification processes to cope with Cd, including the upregulation of genes involved in Cd transport (i.e., absorption, efflux, or compartmentalization), cell wall remodeling, antioxidant system, and chelation. This study provides preliminary information to understand how C. gigantea respond to Cd stress, which is useful for evaluating the potential of C. gigantea in the remediation of Cd-contaminated soils.

The dynamics of cell death patterns and regeneration during acute liver injury in mice.

Acute liver injury is a serious clinical syndrome with multiple causes and unclear pathological process. Here, CCl4 - and D-galactosamine/lipopolysaccharide (D-gal/LPS)-induced acute liver injury was established to explore the cell death patterns and determine whether or not liver regeneration occurred. In CCl4 -induced hepatic injury, three phases, including the early, progressive, and recovery phase, were considered based on alterations of serum transaminases and liver morphology. Moreover, in this model, cytokines exhibited double-peak fluctuations; apoptosis and pyroptosis persisted throughout all phases; autophagy occurred in the early and the progressive phases; and sufficient and timely hepatocyte regeneration was observed only during the recovery phase. All of these phenomena contribute to mild liver injury and subsequent regeneration. Strikingly, only the early and progressive phases were observed in the D-gal/LPS model. Slight pyroptosis occurred in the early phase but diminished in the progressive phase, while apoptosis, reduced autophagy, and slight but subsequently diminished regeneration occurred only during the progressive phase, accompanied by a strong cytokine storm, resulting in severe liver injury with high mortality. Taken together, our work reveals variable modes and dynamics of cell death and regeneration, which lead to different consequences for mild and severe acute liver injury, providing a helpful reference for clinical therapy and prognosis.

The Epigenetic Regulation of OLIG2 by Histone Demethylase KDM6B in Glioma Cells.

Gliomas are common tumors that occur in the brain, accounting for 80% of all malignant brain tumors. Oligodendrocyte transcription factor 2 (OLIG2) is a key transcription factor and strongly expressed in gliomas, which drives proliferation and invasion of glioma cells. Our previous studies have shown that histone lysine (K) demethylase 6B (KDM6B) promotes glioma development. The data also showed that OLIG2 content was positively correlated with KDM6B. Based on this, we proposed that KDM6B may play biological roles by regulating OLIG2 expression. Subsequently, many experiments were performed including specific inhibitor treatment, gene knockdown, and chromatin immunoprecipitation (ChIP) array. These results indicated that inhibition of KDM6B enzymatic activity with GSK-J4 reduces OLIG2 gene expression and protein content. The KDM6B knockdown experiment yielded similar results, that is, it reduces the mRNA and protein level of OLIG2 in glioma cells. ChIP assay showed that the promoter of OLIG2 can be bound by KDM6B, which catalyzes the demethylation of H3K27me3 and increases the expression of OLIG2. This study reveals a new regulatory mechanism of OLIG2 by KDM6B, which has important implications for the future development of drugs for gliomas and other neurological diseases.

A Repair Method for Damage in Aluminum Alloy Structures with the Cold Spray Process.

Aluminum alloy structures may be damaged due to wear or corrosion while in service. These damages will bring about huge financial costs, as well as a huge amount of energy consumption. There is an urgent need to search for an appropriate repair method in order to solve this problem. In this research, the cold spray process was used to repair the damages by using a mixture of powders with Al and Al2O3. A 7N01-T4 aluminum alloy plate with a factitious pit was regarded as the damaged sample. The microstructure, mechanical properties, and corrosion behavior were studied. The results showed that there were no visible perforative pores or cracks in the repaired areas. The microhardness of the repaired areas was in the range of 57.4-63.2 HV and was lower than that of the 7N01-T4 aluminum alloy. The tensile strength of the repaired samples was markedly improved compared with the unrepaired samples. The alternate immersion test results indicated that the repaired samples had the lowest rate of mass loss compared with 7N01-T4 and the unrepaired samples. After alternate immersion tests for 504 h, the repaired samples were covered with dense corrosion products. The repaired samples had a superior corrosion resistance compared to that of 7N01-T4. Thus, the cold spray process is a method of repairing damage in aluminum alloy structures.

Clinical efficacy of mineralized collagen (MC) versus anorganic bovine bone (Bio-Oss) for immediate implant placement in esthetic area: a single-center retrospective study.

BACKGROUND: The purpose of this retrospective study was to evaluate the clinical efficacy of mineralized collagen (MC) versus anorganic bovine bone (Bio-Oss) for immediate implant placement in esthetic area. METHODS: Medical records of Department of Oral and Maxillofacial Surgery of Shandong Provincial Hospital were screened for patients who had been treated with immediate implant implantation in the esthetic area using either MC (Allgens®, Beijing Allgens Medical Science and Technology Co., Ltd., China) or Bio-Oss (Bio-Oss®, Geistlich Biomaterials, Wolhusen, Switzerland), between January 2018 and December 2019. All patients fulfilling the in-/exclusion criteria and following followed for a minimum period of 1 year after surgery were enrolled into the presented study. Implant survival rate, radiographic, esthetic and patient satisfactory evaluations were performed. RESULTS: Altogether, 70 patients were included in the study; a total of 80 implants were inserted. All implants had good initial stability. The survival rate of implants was 100% at 1-year follow-up. The differences in horizontal and vertical bone loss between the MC group (0.72 ± 0.26 mm, 1.62 ± 0.84 mm) and the Bio-Oss group (0.70 ± 0.52 mm, 1.57 ± 0.88 mm) were no significant difference statistically no significant 6 months after permanent restoration. Similar results occurred at 12 months after permanent restoration functional loaded. Clinical acceptability defined by pink esthetic score (PES) ≥ 6 (6.07 ± 1.62 vs. 6.13 ± 1.41) was not significantly different between groups. Patient satisfaction estimated by visual analog scale (VAS) was similar (8.56 ± 1.12 vs. 8.27 ± 1.44), and the difference was no significant difference between the two groups. CONCLUSIONS: The biomimetic MC showed a similar behaviour as Bio-Oss not only in its dimensional tissues changes but also in clinical acceptability and patient satisfaction. Within the limitations of this study, these cases show that MC could be considered as an alternative bone graft in IIP.

Exogenous transforming growth factor-ß1 prevents the inflow of fluoride to ameleoblasts through regulation of voltage-gated chloride channels 5 and 7.

Dental fluorosis is a global issue. Although there are multiple causes of dental fluorosis, the precise mechanism remains controversial. Previous studies have demonstrated that extracellular fluoride may promote an accumulation of fluoride ions in ameloblasts, which may induce oxidative and endoplasmic reticulum stresses, leading to dental fluorosis. However, the exact process by which fluoride ions enter cells has not been determined. In the present study, intracellular fluoride concentration was determined using a newly developed specific fluorescent probe called probe 1. Under high extracellular fluoride concentrations, the fluorescence intensity of the ameloblasts increased, however, exogenous transforming growth factor-ß1 (TGF-ß1) was able to inhibit the increase. Furthermore, changes in the expression of the voltage-gated chloride channels 5 and 7 (ClC5 and ClC-7), which are responsible for the transport of fluoride were investigated. The results indicated that fluoride reduced the expression of endogenous TGF-ß1 and increased the expression of ClC-5 and ClC-7. Additionally, exogenous TGF-ß1 reduced the expression of ClC-5 and ClC-7. The results of the present study indicate that exogenous TGF-ß1 may prevent accumulation of fluoride in ameloblasts through the regulation of ClC-5 and ClC-7 under high extracellular fluoride concentrations.

[Kcnq1ot1 promotes osteogenic differentiation and suppresses osteoclast differentiation].

OBJECTIVE: To investigate the regulatory role of long non-coding RNA Kcnq1ot1 in osteoclast differentiation, osteogenic differentiation and osteoporosis. METHODS: The expression of lnc-Kcnq1ot1, Bglap, Runx2, Alp, Bsp, Nfatc1, Mmp9, Ctsk and Oscar were detected by real-time quantitative PCR (qRT-PCR) in the femoral bones from mouse models of postmenopausal osteoporosis (ovariectomized mice, n=8), disuse osteoporosis (induced by tail suspension, n=14) and agerelated osteoporosis (18-month-old mice, n=8), and also in MC3T3-E1 cells during osteoblast differentiation and in murine bone marrow-derived macrophages (BMMs) and RAW264.7 cells during osteoclast differentiation. MC3T3-E1 cells with lncKcnq1ot1 knockdown by lentivirus infection were induced to differentiate into osteoblasts using osteogenic induction medium, and the expression of lnc-Kcnq1ot1, Alp and Bglap was detected with qRT-PCR and ALP activity was assessed with ALP staining. BMMs and RAW264.7 cells were transfected with siRNAs targeting lnc-Kcnq1ot1 and stimulated with RANKL and/or M-CSF, and the expression of lnc-Kcnq1ot1, Ctsk and Oscar was detected by qRT-PCR, and TRAP activity was assessed by TRAP staining. The subcellular localization of lnc-Kcnq1ot1 in MC3T3-E1 and RAW264.7 cells was determined using cell fractionation followed by qRT-PCR. RESULTS: The expression of lnc-Kcnq1ot1 was significantly upregulated during osteoblast differentiation but downregulated in the bone tissues of osteoporotic mice and during osteoclast differentiation (P < 0.05). Silencing lnc-Kcnq1ot1 obviously decreased the expression of Bglap and Alp (P < 0.05) and attenuated osteogenic medium-induced osteoblast differentiation. Knockdown of lnc-Kcnq1ot1 also promoted the expression of Ctsk and Oscar (P < 0.05) and aggravated RANKL-induced osteoclast differentiation. The results of cell fractionation and qRT-PCR demonstrated that lnc-Kcnq1ot1 was located mainly in the nuclei of MC3T3-E1 and RAW264.7 cells. CONCLUSIONS: Our data demonstrate that lnc-Kcnq1ot1 promotes osteogenic differentiation and alleviates osteoclast differentiation, suggesting the potential of lnc-Kcnq1ot1 as a therapeutic target against osteoporosis.

Potamogeton crispus responses to varying water depth in morphological plasticity and physiological traits.

Submerged macrophytes, important primary producers in shallow lakes, play a crucial role in maintaining ecosystem structure and function. By altering a series of environmental factors, especially light intensity, water depth has great influences on growth of submerged macrophytes. Here, by hanging pots statically at water depths of 40, 60, 80, 100, 120, 140, 160, 180, 200, and 220 cm, respectively, we investigated effects of water depths on morphological plasticity and physiological traits of Potamogeton crispus. At 40 and 60 cm water depths versus other water depths, P. crispus showed lower plant height, larger stem diameter, thicker leaves, and smaller leaf area, leaf length, and specific leaf area. With water depth increasing, the plant height, leaf area, and leaf length gradually increased until 160 cm water depth, while the stem diameter and leaf thickness gradually decreased until 200 cm water depth. In comparison, the plant height, leaf length, and leaf number significantly decreased when the water depth further increased to 180-220 cm. The leaves contained lower concentrations of superoxide dismutase and peroxidase at 100-160 cm water depth, and lower catalase concentrations at 40-140 cm water depth, especially at 80-100 cm. In shallow waters, the concentration of chlorophyll a and b in leaves were both lower, while the ratio of chlorophyll a to b was relatively higher. As the water depth of 40-220 cm, the chlorophyll a and b concentrations increased significantly with increasing water depth, while their ratio gradually decreased. The present study provides new insights into the adaptation strategies of submerged macrophytes to the variation in water levels, and our findings are beneficial for ecosystem construction and management.