Aucubin suppresses TLR4/NF-κB signalling to shift macrophages toward M2 phenotype in glucocorticoid-associated osteonecrosis of the femoral head.

In this study, we investigated whether the ability of aucubin to mitigate the pathology of GONFH involves suppression of TLR4/NF-κB signalling and promotion of macrophage polarization to an M2 phenotype. In necrotic bone tissues from GONFH patients, we compared levels of pro-inflammatory M1 macrophages and anti-inflammatory M2 macrophages as well as levels of TLR4/NF-κB signalling. In a rat model of GONFH, we examined the effects of aucubin on these parameters. We further explored its mechanism of action in a cell culture model of M1 macrophages. Necrotic bone tissues from GONFH patients contained a significantly increased macrophage M1/M2 ratio, and higher levels of TLR4, MYD88 and NF-κB p65 than bone tissues from patients with hip osteoarthritis. Treating GONFH rats with aucubin mitigated bone necrosis and demineralization as well as destruction of trabecular bone and marrow in a dose-dependent manner, based on micro-computed tomography. These therapeutic effects were associated with a decrease in the overall number of macrophages, decrease in the proportion of M1 macrophages, increase in the proportion of M2 macrophages, and downregulation of TLR4, MYD88 and NF-κB p65. These effects in vivo were confirmed by treating cultures of M1 macrophage-like cells with aucubin. Aucubin mitigates bone pathology in GONFH by suppressing TLR4/NF-κB signalling to shift macrophages from a pro- to anti-inflammatory phenotype.

Multi-strain probiotics ameliorate Alzheimer's-like cognitive impairment and pathological changes through the AKT/GSK-3ß pathway in senescence-accelerated mouse prone 8 mice.

BACKGROUND: Alzheimer's disease (AD), the most prevalent type of dementia, still lacks disease-modifying treatment strategies. Recent evidence indicates that maintaining gut microbiota homeostasis plays a crucial role in AD. Targeted regulation of gut microbiota, including probiotics, is anticipated to emerge as a potential approach for AD treatment. However, the efficacy and mechanism of multi-strain probiotics treatment in AD remain unclear. METHODS: In this study, 6-month-old senescence-accelerated-mouse-prone 8 (SAMP8) and senescence-accelerated-mouse-resistant 1 (SAMR1) were utilized. The SAMP8 mice were treated with probiotic-2 (P2, a probiotic mixture of Bifidobacterium lactis and Lactobacillus rhamnosus) and probiotic-3 (P3, a probiotic mixture of Bifidobacterium lactis, Lactobacillus acidophilus, and Lactobacillus rhamnosus) (1 × 109 colony-forming units) once daily for 8 weeks. Morris water maze (MWM) and novel object recognition (NOR) tests were employed to assess the memory ability. 16S sequencing was applied to determine the composition of gut microbiota, along with detecting serum short-chain fatty acids (SCFAs) concentrations. Neural injury, Aß and Tau pathology, and neuroinflammation level were assessed through western blot and immunofluorescence. Finally, potential molecular mechanisms was explored through transcriptomic analysis and western blotting. RESULTS: The MWM and NOR test results indicated a significant improvement in the cognitive level of SAMP8 mice treated with P2 and P3 probiotics compared to the SAMP8 control group. Fecal 16S sequencing revealed an evident difference in the α diversity index between SAMP8 and SAMR1 mice, while the α diversity of SAMP8 mice remained unchanged after P2 and P3 treatment. At the genus level, the relative abundance of ten bacteria differed significantly among the four groups. Multi-strain probiotics treatment could modulate serum SCFAs (valeric acid, isovaleric acid, and hexanoic acid) concentration. Neuropathological results demonstrated a substantial decrease in neural injury, Aß and Tau pathology and neuroinflammation in the brain of SAMP8 mice treated with P3 and P2. Transcriptomic analysis identified the chemokine signaling pathway as the most significantly enriched signaling pathway between SAMP8 and SAMR1 mice. Western blot test indicated a significant change in the phosphorylation level of downstream AKT/GSK-3ß between the SAMP8 and SAMR1 groups, which could be reversed through P2 and P3 treatment. CONCLUSIONS: Multi-strain probiotics treatment can ameliorate cognitive impairment and pathological change in SAMP8 mice, including neural damage, Aß and Tau pathology, and neuroinflammation. This effect is associated with the regulation of the phosphorylation of the AKT/GSK-3ß pathway.

Mechanochemical Synthesis of Cuprous Complexes for X-ray Scintillation and Imaging.

Cuprous complex scintillators show promise for X-ray detection with abundant raw materials, diverse luminescent mechanisms, and adjustable structures. However, their synthesis typically requires a significant amount of organic solvents, which conflict with green chemistry principles. Herein, we present the synthesis of two high-performance cuprous complex scintillators using a simple mechanochemical method for the first time, namely [CuI(PPh3)2R] (R = 4-phenylpyridine hydroiodide (PH, Cu-1) and 4-(4-bromophenyl)pyridine hydroiodide (PH-Br, Cu-2). Both materials demonstrated remarkable scintillation performances, exhibiting radioluminescence (RL) intensities 1.52 times (Cu-1) and 2.52 times (Cu-2) greater than those of Bi4Ge3O12 (BGO), respectively. Compared to Cu-1, the enhanced RL performance of Cu-2 can be ascribed to its elevated quantum yield of 51.54%, significantly surpassing that of Cu-1 at 37.75%. This excellent luminescent performance is derived from the introduction of PH-Br, providing a more diverse array of intermolecular interactions that effectively constrain molecular vibration and rotation, further suppressing the nonradiative transition process. Furthermore, Cu-2 powder can be prepared into scintillator film with excellent X-ray imaging capabilities. This work establishes a pathway for the rapid, eco-friendly, and cost-effective synthesis of high-performance cuprous complex scintillators.

In-MIL-68 derived In2O3/Fe2O3 shuttle-like structures with n-n heterojunctions to improve ethanol sensing performance.

Metal-organic frameworks (MOFs) have a variety of structures and unique properties that make them suitable for use in gas sensors. Herein, In2O3/Fe2O3 was successfully synthesized using simple solvothermal and impregnation methods. The response to 100 ppm of ethanol gas reached 67.5 at an optimum working temperature of 200 °C, and the response/recovery time was 9 s/236 s. The composite also exhibited excellent selectivity, repeatability, and long-term stability. SEM, TEM, XRD, and XPS were used for the characterization of materials. The excellent sensing performance of the sensors is attributed to the construction of n-n heterojunctions, an increase in oxygen vacancies, and the unique structural characteristics of MOFs. The above experimental results indicate that In-MIL-68-derived In2O3/Fe2O3 is a promising ethanol sensing material.

Association between history of cannabis use and outcomes after total hip or knee arthroplasty: a systematic review and meta-analysis.

Background: Cannabis use may be increasing as countries legalize it and it becomes socially acceptable. A history of cannabis use may increase risk of complications after various kinds of surgery and compromise functional recovery. Here we systematically reviewed and meta-analyzed available evidence on how history of cannabis use affects recovery after hip or knee arthroplasty (THA/TKA). Methods: The PubMed, EMBASE, and Web of Science databases were comprehensively searched and studies were selected and analyzed in accordance with the PRISMA guidelines. The methodological quality of included studies was assessed based on the Newcastle-Ottawa Scale, while quality of evidence was evaluated according to the "Grading of recommendations assessment, development, and evaluation" system. Data on various outcomes were pooled when appropriate and meta-analyzed. Results: The systematic review included 16 cohort studies involving 5.91 million patients. Meta-analysis linked history of cannabis use to higher risk of the following outcomes: revision (RR 1.68, 95% CI 1.31-2.16), mechanical loosening (RR 1.77, 95% CI 1.52-2.07), periprosthetic fracture (RR 1.85, 95% CI 1.38-2.48), dislocation (RR 2.10, 95% CI 1.18-3.73), cardiovascular events (RR 2.49, 95% CI 1.22-5.08), cerebrovascular events (RR 3.15, 95% CI 2.54-3.91), pneumonia (RR 3.97, 95% CI 3.49-4.51), respiratory failure (RR 4.10, 95% CI 3.38-4.97), urinary tract infection (RR 2.46, 95% CI 1.84-3.28), acute kidney injury (RR 3.25, 95% CI 2.94-3.60), venous thromboembolism (RR 1.48, 95% CI 1.34-1.63), and deep vein thrombosis (RR 1.42, 95% CI 1.19-1.70). In addition, cannabis use was associated with significantly greater risk of postoperative transfusion (RR 2.23, 95% CI 1.83-2.71) as well as higher hospitalization costs. Conclusion: History of cannabis use significantly increases the risk of numerous complications and transfusion after THA or TKA, leading to greater healthcare costs. Clinicians should consider these factors when treating cannabis users, and pre-surgical protocols should give special consideration to patients with history of cannbis use.

Altered gut microbe metabolites in patients with alcohol­induced osteonecrosis of the femoral head: An integrated omics analysis.

Excessive alcohol consumption is considered to be a major risk factor of alcohol-induced osteonecrosis of the femoral head (AONFH). The gut microbiota (GM) has been reported to aid in the regulation of human physiology and its composition can be altered by alcohol consumption. The aim of the present study was to improve the understanding of the GM and its metabolites in patients with AONFH. Metabolomic sequencing and 16S rDNA analysis of fecal samples were performed using liquid chromatography-mass spectrometry to characterize the GM of patients with AONFH and healthy normal controls (NCs). Metagenomic sequencing of fecal samples was performed to identify whether GM changes on the species level were associated with the expression of gut bacteria genes or their associated functions in patients with AONFH. The abundance of 58 genera was found to differ between the NC group and the AONFH group. Specifically, Klebsiella, Holdemanella, Citrobacter and Lentilactobacillus were significantly more abundant in the AONFH group compared with those in the NC group. Metagenomic sequencing demonstrated that the majority of the bacterial species that exhibited significantly different abundance in patients with AONFH belonged to the genus Pseudomonas. Fecal metabolomic analysis demonstrated that several metabolites were present at significantly different concentrations in the AONFH group compared with those in the NC group. These metabolites were products of vitamin B6 metabolism, retinol metabolism, pentose and glucuronate interconversions and glycerophospholipid metabolism. In addition, these changes in metabolite levels were observed to be associated with the altered abundance of specific bacterial species, such as Basidiobolus, Mortierella, Phanerochaete and Ceratobasidium. According to the results of the present study, a comprehensive landscape of the GM and metabolites in patients with AONFH was revealed, suggesting the existence of interplay between the gut microbiome and metabolome in AONFH pathogenesis.

MXene/Silk Fibroin Strengthened PVA-Based Eutectogel with Excellent Self-Healing Ability and Environmental Adaptability: Design, Synthesis, and Sensing Application.

A superelastic self-healing eutectogel was designed and prepared using poly (vinyl alcohol) (PVA) as the bulk skeleton material, while silk fibroin (SF) and two-dimensional (2D) MXene (Ti3C2TX) as reinforcing fillers. In brief, the eutectogel possesses a high tensile strength of 7.63 MPa, and its elongation at break reached 1115.2%, higher than most reported polymers (<1000%). In addition, the eutectogel-assembled sensor has a high ionic conductivity of 0.61 S/m and a high strain sensitivity of 5.17 kPa-1. Moreover, eutectogel shows excellent self-healing ability and can achieve self-healing quickly within 10 min, while its tensile strength and elongation at break can be restored to 84.7% and 97.4% of the initial levels. Besides, a stable electrical signal can be transmitted after 200 cycles at 30% strain. Finally, the eutectogel can withstand various environmental conditions, such as atmospheric or even vacuum evaporation and low-temperature freezing, while maintaining good mechanical and sensing performances. The assembled flexible sensors based on the eutectogel demonstrate their significant application prospects in wearable devices, especially human physiological monitoring.

Structural insights into the orthosteric inhibition of P2X receptors by non-ATP analog antagonists.

P2X receptors are extracellular ATP-gated ion channels that form homo- or heterotrimers and consist of seven subtypes. They are expressed in various tissues, including neuronal and nonneuronal cells, and play critical roles in physiological processes such as neurotransmission, inflammation, pain, and cancer. As a result, P2X receptors have attracted considerable interest as drug targets, and various competitive inhibitors have been developed. However, although several P2X receptor structures from different subtypes have been reported, the limited structural information of P2X receptors in complex with competitive antagonists hampers the understanding of orthosteric inhibition, hindering the further design and optimization of those antagonists for drug discovery. We determined the cryogenic electron microscopy (cryo-EM) structures of the mammalian P2X7 receptor in complex with two classical competitive antagonists of pyridoxal-5'-phosphate derivatives, pyridoxal-5'-phosphate-6-(2'-naphthylazo-6'-nitro-4',8'-disulfonate) (PPNDS) and pyridoxal phosphate-6-azophenyl-2',5'-disulfonic acid (PPADS), and performed structure-based mutational analysis by patch-clamp recording as well as molecular dynamics (MD) simulations. Our structures revealed the orthosteric site for PPADS/PPNDS, and structural comparison with the previously reported apo- and ATP-bound structures showed how PPADS/PPNDS binding inhibits the conformational changes associated with channel activation. In addition, structure-based mutational analysis identified key residues involved in the PPNDS sensitivity of P2X1 and P2X3, which are known to have higher affinity for PPADS/PPNDS than other P2X subtypes.

Comparative effects of progestin-based combination therapy for endometrial cancer or atypical endometrial hyperplasia: a systematic review and network meta-analysis.

Objectives: The objective of this network meta-analysis is to systematically compare the efficacy of diverse progestin-based combination regimens in treating patients diagnosed with endometrial cancer or atypical endometrial hyperplasia. The primary goal is to discern the optimal combination treatment regimen through a comprehensive examination of their respective effectiveness. Methods: We systematically searched four prominent databases: PubMed, Web of Science, Embase, and Cochrane Central Register of Controlled Trials, for randomized controlled trials addressing the efficacy of progestins or progestin combinations in the treatment of patients with endometrial cancer or atypical endometrial hyperplasia. The search spanned from the inception of these databases to December 2023. Key outcome indicators encompassed survival indices, criteria for assessing efficacy, as well as pregnancy and relapse rate. This study was registered in PROSPERO (CRD42024496311). Results: From the 1,558 articles initially retrieved, we included 27 studies involving a total of 5,323 subjects in our analysis. The results of the network meta-analysis revealed that the mTOR inhibitor+megestrol acetate (MA)+tamoxifen regimen secured the top rank in maintaining stable disease (SD) (SUCRA=73.4%) and extending progression-free survival (PFS) (SUCRA=72.4%). Additionally, the progestin combined with tamoxifen regimen claimed the leading position in enhancing the partial response (PR) (SUCRA=75.2%) and prolonging overall survival (OS) (SUCRA=80%). The LNG-IUS-based dual progestin regimen emerged as the frontrunner in improving the complete response (CR) (SUCRA=98.7%), objective response rate (ORR) (SUCRA=99.1%), pregnancy rate (SUCRA=83.7%), and mitigating progression (SUCRA=8.0%) and relapse rate (SUCRA=47.4%). In terms of safety, The LNG-IUS-based dual progestin regimen had the lowest likelihood of adverse events (SUCRA=4.2%), while the mTOR inhibitor regimen (SUCRA=89.2%) and mTOR inbitor+MA+tamoxifen regimen (SUCRA=88.4%) had the highest likelihood of adverse events. Conclusions: Patients diagnosed with endometrial cancer or atypical endometrial hyperplasia exhibited the most favorable prognosis when undergoing progestin combination therapy that included tamoxifen, mTOR inhibitor, or LNG-IUS. Notably, among these options, the LNG-IUS-based dual progestin regimen emerged as particularly promising for potential application. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO, identifier CRD42024496311.

Polyoxometalate/s-triazine hybrid heterostructures with ultrafast photochromic properties.

As an emerging class of hybrid complexes, donor-acceptor (D-A) hybrid heterostructures, which combine the advantages of both organic and inorganic photoactive components, provide excellent platforms for the fabrication of photochromic materials with enhanced photo-responsive performances. Herein, four novel hybrid heterostructures, namely H3TPT·(PW12O40)·2NMP (1), (H1.5TPT)2·(PW12O40) (2), (H3TPT)2·(SiW12O40)·2Cl·2MeCN (3), and H3TPT·(HPMo12O40)·Cl·3NMP (4) (TPT is tri(4-pyridyl)-s-triazine, NMP is N-methylpyrrolidone), have been synthesized and characterized. Benefitting from the strong interactions (anion-π interactions) and matching electron energy levels between the donors and acceptors, some of them exhibited ultrafast photochromic behaviour even up to 1 second. Furthermore, based on experimental and theoretical calculations, the plausible PIET process and structure-activity relationship have been discussed in detail.

A shared mechanism for TNP-ATP recognition by members of the P2X receptor family.

P2X receptors (P2X1-7) are non-selective cation channels involved in many physiological activities such as synaptic transmission, immunological modulation, and cardiovascular function. These receptors share a conserved mechanism to sense extracellular ATP. TNP-ATP is an ATP derivative acting as a nonselective competitive P2X antagonist. Understanding how it occupies the orthosteric site in the absence of agonism may help reveal the key allostery during P2X gating. However, TNP-ATP/P2X complexes (TNP-ATP/human P2X3 (hP2X3) and TNP-ATP/chicken P2X7 (ckP2X7)) with distinct conformations and different mechanisms of action have been proposed. Whether these represent species and subtype variations or experimental differences remains unclear. Here, we show that a common mechanism of TNP-ATP recognition exists for the P2X family members by combining enhanced conformation sampling, engineered disulfide bond analysis, and covalent occupancy. In this model, the polar triphosphate moiety of TNP-ATP interacts with the orthosteric site, while its TNP-moiety is deeply embedded in the head and dorsal fin (DF) interface, creating a restrictive allostery in these two domains that results in a partly enlarged yet ion-impermeable pore. Similar results were obtained from multiple P2X subtypes of different species, including ckP2X7, hP2X3, rat P2X2 (rP2X2), and human P2X1 (hP2X1). Thus, TNP-ATP uses a common mechanism for P2X recognition and modulation by restricting the movements of the head and DF domains which are essential for P2X activation. This knowledge is applicable to the development of new P2X inhibitors.

A Clinical Breathomics Dataset.

This study entailed a comprehensive GC‒MS analysis conducted on 121 patient samples to generate a clinical breathomics dataset. Breath molecules, indicative of diverse conditions such as psychological and pathological states and the microbiome, were of particular interest due to their non-invasive nature. The highlighted noninvasive approach for detecting these breath molecules significantly enhances diagnostic and monitoring capacities. This dataset cataloged volatile organic compounds (VOCs) from the breath of individuals with asthma, bronchiectasis, and chronic obstructive pulmonary disease. Uniform and consistent sample collection protocols were strictly adhered to during the accumulation of this extensive dataset, ensuring its reliability. It encapsulates extensive human clinical breath molecule data pertinent to three specific diseases. This consequential clinical breathomics dataset is a crucial resource for researchers and clinicians in identifying and exploring important compounds within the patient's breath, thereby augmenting future diagnostic and therapeutic initiatives.

Forcefully engaging rods into tulips with gap discrepancy leading to pedicle screw loosening-a biomechanical analysis using long porcine spine segments.

BACKGROUND CONTEXT: Long-segment pedicle screw instrumentation is widely used to treat complex spinal disorders. Rods are routinely precontoured to maximize assistance on the correcting side of the deformity, but there often exists a residual gap discrepancy between the precontoured rods and screw tulips. No previous research has investigated the diminished pullout strength of the most proximal or distal pedicle screw resulting from a mismatched rod in long-segment pedicle screw instrumentation. PURPOSE: The present study aimed to investigate the decreased pullout force of pedicle screws affected by the gap discrepancy when forcefully engaging a mismatched rod into a tulip in a normal-density porcine spine. STUDY DESIGN: The pedicle screw fixation strength under axial pullout force was compared among three different gap discrepancies between rods and tulips using long porcine spine segments. METHODS: Twelve porcine lumbar vertebrae (L3-L6) were implanted with pedicle screws and rods. Screws on one side had no gap between the tulip and rod (0-mm group), while the most proximal screw on the other side had an intentional gap of 3 mm (3-mm group) or 6 mm (6-mm group). Three hours after forcefully engaging the rod into the tulips at room temperature, the set screws in all specimens were loosened, and each specimen was dissected into individual vertebrae for subsequent pullout testing. RESULTS: The control group exhibited significantly greater pullout strength (1987.68 ± 126.80 N) than the groups from different rod-tulip configurations (p<.05), with significantly greater strength in the 3-mm group (945.62 ± 97.43 N) than the 6-mm group (655.30 ± 194.49 N) (p<.05). Only 47.6% and 33.0% of the pullout strength was retained in the 3-mm and 6-mm groups, respectively, compared to the control group. CONCLUSIONS: Gap discrepancies between rods and tulips can significantly reduce pedicle screw pullout strength, with a correlation between decreased strength and increased gaps. Surgeons should avoid forcefully engaging mismatched rods and consider well-fitted contoured rods in spinal surgery to minimize the risk of screw loosening. CLINICAL SIGNIFICANCE: The gap discrepancy between rod and tulip significantly affected pullout strength, with greater gaps leading to reduced strength. Forcefully engaging mismatched rods into tulips in degenerative spinal surgery should be avoided to minimize the risk of early screw pullout.

Skeletal muscle index together with body mass index is associated with secondary osteoporosis in patients with rheumatoid arthritis.

OBJECTIVE: The objective of this study was to explore the associations of body mass index (BMI), fat mass index (FMI), skeletal mass index (SMI) and secondary osteoporosis (OP) in patients with rheumatoid arthritis (RA). METHODS: The bone mineral density (BMD) at sites of the femur neck (Neck), total hip (Hip) and lumbar vertebrae 1-4 (L1-4) was measured by dual-energy X-ray absorptiometry. The skeletal muscle index, body fat percentage and mineral content were measured by biological electrical impedance for calculating BMI, FMI and SMI. RESULTS: A total of 433 patient with RA and 158 healthy controls were enrolled. The BMDs at each site of the RA patients were lower compared with those of the healthy controls (p < 0.0001), and the prevalence of OP (36.1%, 160/443) and sarcopenia (65.2%, 288/443) in the RA patients were higher than those in the controls (12.7%, 20/158, p < 0.0001; 9.0%, 14/156, p < 0.0001). Significant differences in the BMD, FMI, SMI, mineral content, body fat percentage and skeletal muscle mass were found among the RA patients in the different BMI groups (p < 0.05). In RA patients with BMI < 18.5 kg/m2, the prevalence of OP in the RA patients with sarcopenia was similar to that in those without sarcopenia (44.4% vs. 66. 7%, χ2 = 0. 574, p = 0.449). In the RA patients with a normal BMI or who were overweight or obese, prevalence of OP in the RA patients with sarcopenia was significantly higher than that in the RA patients without sarcopenia (42.8% vs. 21.7%, χ2 = 10.951, p = 0.001; 61.1% vs. 13.0%, χ2 = 26.270, p < 0.0001). In the RA patients without sarcopenia, the prevalence of OP in the RA patients in the different BMI groups was different (p = 0.039). In the RA patients with sarcopenia, there was no significant difference in the prevalence of OP among the RA patients in the different BMI groups (p = 0. 128). The linear correlation analysis showed that the SMI in RA patients was positively correlated with the BMD of each site measured and BMI and FMI (p < 0.0001). However, there was a negative linear correlation between SMI and disease duration (p = 0.048). The logistic regression analysis found that SMI (OR = 0.569, p = 0.002, 95% CI 0.399-0.810), BMI (OR = 0.884, p = 0.01, 95% CI 0.805-0.971) and gender (1 = female, 2 = male) (OR = 0.097, p < 0.0001, 95% CI 0.040-0.236) were protective factors for OP in RA, while age (OR = 1.098, p < 0.0001, 95% CI 1.071-1.125) was the risk factor. CONCLUSION: BMI and SMI are associated with the occurrence of OP in RA patients, and both SMI and BMI are important protective factors for OP secondary to RA.

Mir-142-3P regulates MAPK protein family by inhibiting 14-3-3η to enhance bone marrow mesenchymal stem cells osteogenesis.

Clinical studies have found 14-3-3η to be associated with osteoporosis through undefined mechanisms. We aimed to investigate the role of 14-3-3η in osteoporosis and its potential associations with miRNAs. The Gene Expression Omnibus(GEO) and Human Protein Atlas 1 databases were analyzed to examine both the mRNA and protein expression of 14-3-3η in OP. Gene enrichment analyses were performed to explore the underlying mechanism of 14-3-3η based on DAVID. miRWalk was used to predict the associated miRNAs. The statistics were analysed by R software and SPSS software. 14-3-3η was overexpressed and knock down expressed in BMSCs by lentiviral vector transfecting. And BMSCs were induced by hypoxia. qRT-PCR and Western-Blot verified the expression of mRNA and protein. Scratch assay detected the migration of osteocytes. Co-immunoprecipitation and luciferase assay studied the 14-3-3η targeted protein and miRNA. overexpression and knock down of miRNA to verify the relationship of 14-3-3η and target genes. The 14-3-3η mRNA expression level was low in patients with osteoporosis, as corroborated by immunohistochemical staining images. Functional analyses revealed enrichment of the MAPK-associated cascade. 14-3-3η was correlated with MAPK family proteins and five key miRNAs, including mir-142-3p. In addition, 14-3-3η knockdown in BMSCs increased the mRNA and protein expression levels of Hif-α, VEGF, BMP-2, OPN, OST, and Runx2, and enhanced the cells migration ability. Under hypoxic conditions, Hif-α and BMP-2 protein expression levels were upregulated, whereas those of 14-3-3η and MAPK3 were downregulated. Co-immunoprecipitation experiments showed decreased binding of 14-3-3η to MAPK3. 14-3-3η knockdown produced the same results as hypoxia induction. Adding caspase3 inhibitor and knocking down 14-3-3η again prevented MAPK3 cleavage by caspase3 and inhibited BMP-2 expression. Moreover, under hypoxic conditions, miR-142-3P expression was upregulated and luciferase assays revealed 14-3-3η as its target gene. miR-142-3P overexpression decreased mRNA and protein levels of 14-3-3η and MAPK3, while increasing BMP-2 expression. miR-142-3P knockdown reversed these results. BMSC osteogenesis was suppressed by 14-3-3η, whereas miRNA-142-3p promoted it through the inhibition of 14-3-3η.

Identification of the myodural bridge in a venomous snake, the gloydius shedaoensis: what is the functional significance? / Identificación del puente miodural en una serpiente venenosa, gloydius shedaoensis: ¿cuál es el significado funcional?

SUMMARY: Myodural bridges (MDB) are anatomical connections between the suboccipital muscles and the cervical dura mater which pass through both the atlanto-occipital and the atlanto-axial interspaces in mammals. In our previous studies, we found that the MDB exists in seven terrestrial mammal species, two marine mammal species, two reptilian species, and one bird species. A recent study suggested that given the "ubiquity" of myodural bridges in terrestrial vertebrates, the MDB may also exist in snakes. Specifically, we focused on the Gloydius shedaoensis, a species of Agkistrodon (pit viper snake) that is only found on Shedao Island, which is in the southeastern sea of Dalian City in China. Six head and neck cadaveric specimens of Gloydius shedaoensis were examined. Three specimens were used for anatomical dissection and the remaining three cadaveric specimens were utilized for histological analysis. The present study confirmed the existence of the MDB in the Gloydius shedaoensis. The snake's spinalis muscles originated from the posterior edge of the supraoccipital bones and the dorsal facet of the exocciput, and then extended on both sides of the spinous processes of the spine, merging with the semispinalis muscles. On the ventral aspect of this muscular complex, it gave off fibers of the MDB. These MDB fibers twisted around the posterior margin of the exocciput and then passed through the atlanto-occipital interspace, finally terminating on the dura mater. We observed that the MDB also existed in all of the snakes' intervertebral joints. These same histological findings were also observed in the Gloydius brevicaudus, which was used as a control specimen for the Gloydius shedaoensis. In snakes the spinal canal is longer than that observed in most other animals. Considering the unique locomotive style of snakes, our findings contribute to support the hypothesis that the MDB could modulate cerebrospinal fluid (CSF) pulsations.

RESUMEN: Los puentes miodurales (MDB) son conexiones anatómicas entre los músculos suboccipitales y la duramadre cervical que pasan a través de los espacios intermedios atlanto-occipital y atlanto-axial en los mamíferos. En nuestros estudios anteriores, encontramos que el MDB existe en siete especies de mamíferos terrestres, dos especies de mamíferos marinos, dos especies de reptiles y una especie de ave. Un estudio reciente sugirió que dada la "ubicuidad" de los puentes miodurales en los vertebrados terrestres, el MDB también puede existir en las serpientes. Específicamente, nos enfocamos en Gloydius shedaoensis, una especie de Agkistrodon (serpiente víbora) que solo se encuentra en la isla Shedao, en el mar sureste de la ciudad de Dalian en China. Se examinaron seis especímenes cadavéricos de cabeza y cuello de Gloydius shedaoensis. Se utilizaron tres especímenes para la disección anatómica y los tres especímenes cadavéricos restantes se utilizaron para el análisis histológico. El presente estudio confirmó la existencia del MDB en Gloydius shedaoensis. Los músculos espinosos de la serpiente se originaron en el margen posterior de los huesos supraoccipital y la cara dorsal del exoccipucio, y luego se extendieron a ambos lados de los procesos espinosas de la columna vertebral, fusionándose con los músculos semiespinosos. En la cara ventral de este complejo muscular se desprendían fibras del MDB. Estas fibras MDB se ubican alrededor del margen posterior del exoccipucio y luego atraviesan el interespacio atlanto-occipital, terminando finalmente en la duramadre. Observamos que el MDB también existía en todas las articulaciones intervertebrales de las serpientes. Estos mismos hallazgos histológicos también se observaron en Gloydius brevicaudus, que se utilizó como muestra de control para Gloydius shedaoensis. En las serpientes, el canal espinal es más largo que el observado en la mayoría de los otros animales. Teniendo en cuenta el estilo único locomotor de las serpientes, nuestros hallazgos contribuyen a respaldar la hipótesis de que el MDB podría modular las pulsaciones del líquido cerebroespinal.

Morphological index of fetal cerebral ventricular development: a diagnostic ultrasound study / Índice morfológico del desarrollo ventricular cerebral fetal: un estudio ecográfico diagnóstico

SUMMARY: To establish an unprovable diagnostic indicative index reference for ultrasound examination of the fetal cerebral ventricles, based on the morphological characteristics throughout fetal nervous system development. Key ultrasonic morphological indicators of fetal ventricular development, which includes frontal horn width (FHW), occipital horn width (OHW), width of 3rd ventricle, cavity of septum pellucidum (CSP), width and length of 4th ventricle and thalamo-occipital distance (TOD) had been measured and analyzed collectively. All data of the indicators was collected on singleton pregnant woman between 16-39 weeks of gestational age (GA), between November 2017 and June 2021 at the Second Hospital of Dalian Medical University. A total of 235 pregnant women were enrolled in the cross section study; another 36 pregnant women voluntarily joined a timeline-tracking follow-up study (cohort study) under the same examining criteria. A decrease of FHW and OHW of the lateral ventricles was observed as GA increased; while dimensional values of TOD, 3rd ventricle, CSP, as well as 4th ventricle increased with GA. Most of these indicators showed an enhanced variation tendency within a certain period of GA. Moreover, values of FHW and TOD showed asymmetry of the two hemispheres within the whole GA. Our findings revealed the morphological regularity of fetal ventricular development, which would instructively enhance the relative clinical ultrasound diagnosis; moreover, TOD also showed regularly changes as GA increased, suggesting that TOD should be considered as an additional routine ultrasonic indicator for fetal ventricular development.

RESUMEN: El objetivo del estudio fue establecer un índice de referencia indicativo diagnóstico no demostrable para el examen ecográfico de los ventrículos cerebrales fetales, basado en las características morfológicas a lo largo del desarrollo del sistema nervioso fetal. Indicadores morfológicos ultrasónicos clave del desarrollo ventricular fetal, que incluyen el ancho del cuerno frontal (FHW), el ancho del cuerno occipital (OHW), el ancho del tercer ventrículo, la cavidad del septo pelúcido (CSP), el ancho y el largo del cuarto ventrículo y la distancia tálamo-occipital (TOD) fueron medidos y analizados conjuntamente. Todos los datos de los indicadores se recopilaron en mujeres embarazadas de un solo feto entre 16 y 39 semanas de edad gestacional (EG), entre noviembre de 2017 y junio de 2021 en el Segundo Hospital de la Universidad Médica de Dalian. Un total de 235 mujeres embarazadas se inscribieron en el estudio transversal; otras 36 mujeres embarazadas se unieron voluntariamente a un estudio de seguimiento de línea de tiempo (estudio de cohorte) bajo los mismos criterios de examen. Se observó una disminución de FHW y OHW de los ventrículos laterales a medida que aumentaba la GA; mientras que los valores dimensionales de TOD, tercer ventrículo, CSP y cuarto ventrículo aumentaron con GA. La mayoría de estos indicadores mostraron una tendencia de variación mejorada dentro de un cierto período de GA. Además, los valores de FHW y TOD mostraron asimetría de los dos hemisferios dentro de toda la AG. Nuestros hallazgos revelaron la regularidad morfológica del desarrollo ventricular fetal, lo que mejoraría de manera instructiva el diagnóstico clínico de ultrasonido relativo; además, TOD también mostró cambios regulares a medida que aumentaba la GA, lo que sugiere que TOD debe considerarse como un indicador ultrasónico de rutina adicional para el desarrollo ventricular fetal.