Clinical and gonadal transcriptome analysis of 38,XX disorder of sex development pigs.

Pigs serve as a robust animal model for the study of human diseases, notably in the context of disorders of sex development (DSD). This study aims to investigate the phenotypic characteristics and molecular mechanisms underlying the reproductive and developmental abnormalities of 38,XX ovotestis-DSD (OT-DSD) and 38,XX testis-DSD (T-DSD) in pigs. Clinical and transcriptome sequencing analyses were performed on DSD and normal female pigs. Cytogenetic and SRY analyses confirmed that OT/T-DSD pigs exhibited a 38,XX karyotype and lacked the SRY gene. The DSD pigs had higher levels of follicle-stimulating hormone, luteinizing hormone, and progesterone, but lower testosterone levels when compared with normal male pigs. The reproductive organs of OT/T-DSD pigs exhibit abnormal development, displaying both male and female characteristics, with an absence of germ cells in the seminiferous tubules. Sex determination and development-related differentially-expressed genes (DEGs) shared between DSD pigs were identified in the gonads, including WT1, DKK1, CTNNB1, WTN9B, SHOC, PTPN11, NRG1 and NXK3-1. DKK1 is proposed as a candidate gene for investigating the regulatory mechanisms underlying gonadal phenotypic differences between OT-DSD and T-DSD pigs. Consequently, our findings provide insights into the molecular pathogenesis of DSD pigs and present an animal model for studying into DSD in humans.

Decoding the transcriptome of muscular dystrophy due to Ptrf deficiency using single-nucleus RNA sequencing.

Lacking PTRF (polymerase I and transcript release factor), an essential caveolae component, causes a secondary deficiency of caveolins resulting in muscular dystrophy. The transcriptome responses of different types of muscle fibers and mononuclear cells in skeletal muscle to muscular dystrophy caused by Ptrf deletion have not been explored. Here, we created muscular dystrophy mice by Ptrf knockout and applied single-nucleus RNA sequencing (snRNA-seq) to unveil the transcriptional changes of the skeletal muscle at single-nucleus resolution. 11 613 muscle nuclei (WT, 5838; Ptrf KO, 5775) were classified into 12 clusters corresponding to 11 nuclear types. Trajectory analysis revealed the potential transition between type IIb_1 and IIb_2 myonuclei upon muscular dystrophy. Functional enrichment analysis indicated that apoptotic signaling and enzyme-linked receptor protein signaling pathway were significantly enriched in type IIb_1 and IIb_2 myonuclei of Ptrf KO, respectively. The muscle structure development and the PI3K-AKT signaling pathway were significantly enriched in type IIa and IIx myonuclei of Ptrf KO. Meanwhile, metabolic pathway analysis showed a decrease in overall metabolic pathway activity of myonuclei subtypes upon muscular dystrophy, with the most decrease in type IIb_1 myonuclei. Gene regulatory network analysis found that the activity of Mef2c, Mef2d, Myf5, and Pax3 regulons was enhanced in type II myonuclei of Ptrf KO, especially in type IIb_2 myonuclei. In addition, we investigated the transcriptome changes in adipocytes and found that muscular dystrophy enhanced the lipid metabolic capacity of adipocytes. Our findings provide a valuable resource for exploring the molecular mechanism of muscular dystrophy due to Ptrf deficiency.

The horizontal gene transfer of perchlorate reduction genomic island in three bacteria from an ecological niche.

Three new strains of dissimilatory perchlorate-reducing bacteria (DPRB), QD19-16, QD1-5, and P3-1, were isolated from an active sludge. Phylogenetic trees based on 16S rRNA genes indicated that QD19-16, QD1-5, and P3-1 belonged to Brucella, Acidovorax, and Citrobacter, respectively, expanding the distribution of DPRB in the Proteobacteria. The three strains were gram-negative and facultative anaerobes with rod-shaped cells without flagella, which were 1.0-1.6 µm long and 0.5-0.6 µm wide. The three DPRB strains utilized similar broad spectrum of electron donors and acceptors and demonstrated a similar capability to reduce perchlorate within 6 days. The enzyme activity of perchlorate reductase in QD19-16 toward chlorate was higher than that toward perchlorate. The high sequence similarity of the perchlorate reductase operon and chlorite dismutase genes in the perchlorate reduction genomic islands (PRI) of the three strains implied that they were monophyletic origin from a common ancestral PRI. Two transposase genes (tnp1 and tnp2) were found in the PRIs of strain QD19-16 and QD1-5, but were absent in the strain P3-1 PRI. The presence of fragments of IR sequences in the P3-1 PRI suggested that P3-1 PRI had previously contained these two tnp genes. Therefore, it is plausible to suggest that a common ancestral PRI transferred across the strains Brucella sp. QD19-16, Acidovorax sp. QD1-5, and Citrobacter sp. P3-1 through horizontal gene transfer, facilitated by transposases. These results provided a direct evidence of horizontal gene transfer of PRI that could jump across phylogenetically unrelated bacteria through transposase. KEY POINTS: • Three new DPRB strains can effectively remove high concentration of perchlorate. • The PRIs of three DPRB strains are acquired from a single ancestral PRI. • PRIs are incorporated into different bacteria genome through HGT by transposase.

Effects of the promoting bacterium on growth of plant under cadmium stress.

Cadmium (Cd) pollution is a huge threat to ecosystem health. In the manuscript, pot experiments were conducted to investigate the changes in plant biomass and antioxidant indicators under different cadmium pollution levels (0, 25, 50, and 100 mg/kg) of inoculation of plant growth-promoting bacteria ZG7 on sugar beet. The results showed that the accumulation of excess Cd in sugar beet exhibited different symptoms, including reduced biomass (p < 0.05). Compared with the group treated with uninoculated strain ZG7, inoculation of strain ZG7 significantly reduced the toxicity of sugar beet to Cd and enhanced its antioxidant capacity, with no significant differences in root biomass and increases in leaf biomass of 15.71, 5.84, and 74.12 under different Cd concentration treatments (25, 50, and 100 mg/kg), respectively. The root enrichment of Cd was reduced by 49.13, 47.26, and 21.50%, respectively (p < 0.05). The leaf fraction was reduced by 59.35, 29.86, and 30.99%, respectively (p < 0.05). In addition, the enzymatic activities of sucrase, urease, catalase, and neutral phosphatase were significantly enhanced in the soil (p < 0.05). This study helps us to further investigate the mechanism of cadmium toxicity reduction by inoculated microorganisms and provides a theoretical reference for growing plants in cadmium-contaminated agricultural fields.

The combination of microorganisms and phytoremediation is becoming a popular research topic. The selection of suitable plant growth promoting bacteria can not only promote the growth and development of plants and enhance their cadmium resistance, but also improve the soil quality. And the results of this study will be important for growing edible plants and improving soils in cadmium-contaminated areas.

A nomogram for predicting choledochal cyst with perforation.

BACKGROUND: Choledochal cyst with perforation (CC with perforation) rarely occurs, early diagnosis and timely treatment plan are crucial for the treatment of CC with perforation. This study aims to forecast the occurrence of CC with perforation. METHODS: All 1111 patients were conducted, who underwent surgery for choledochal cyst at our hospital from January 2011 to October 2022. We conducted univariate and multivariate logistic regression analysis to screen for independent predictive factors for predicting CC with perforation, upon which established a nomogram. The predictive performance of the nomogram was evaluated using receiver operating characteristic (ROC) curves, calibration plots, and decision curve analysis (DCA) curves. RESULTS: The age of children with choledochal cyst perforation is mainly concentrated between 1 and 3 years old. Logistic regression analysis indicates that age, alanine aminotransferase, glutamyl transpeptidase, C-reactive protein, vomiting, jaundice, abdominal distension, and diarrhea are associated with predicting the occurrence of choledochal cyst perforation. ROC curves, calibration plots, and DCA curve analysis curves demonstrate that the nomogram has great discriminative ability and calibration, as well as significant clinical utility. CONCLUSION: The age of CC with perforation is mainly concentrated between 1 and 3 years old. A nomogram for predicting the perforation of choledochal cyst was established.

Inoculation of Exogenous Complex Bacteria to Enhance Resistance in Alfalfa and Combined Remediation of Heavy Metal-Contaminated Soil.

Heavy metals are considered to be one of the main sources of soil contamination. In this study, three tolerant bacteria were isolated from the heavy metal-contaminated soil in mining area, and immobilized bacteria were constructed using corn straw as the carrier. The combined remediation effect of immobilized bacteria and alfalfa in pot experiments was explored in heavy metal-contaminated soil. Under heavy metal stress, inoculation with immobilized bacteria significantly promoted the growth of alfalfa, in which the dry weights of roots, stems, and leaves increased by 19.8, 6.89, and 14.6%, respectively (P < 0.05). Also, inoculation with immobilized bacteria improved the antioxidant capacity of plants and the activity of soil enzymes and improved soil quality (P < 0.05). Microbial-phytoremediation technology effectively reduced the heavy metal content in the soil, and can restore the soil contaminated by heavy metals. The results will help to further understand the mechanism of microbial inoculation to reduce the toxicity of heavy metals, and provide guidance for the cultivation of forage grasses in heavy metal-contaminated soils.

Inpainting Saturation Artifact in Anterior Segment Optical Coherence Tomography.

The cornea is an important refractive structure in the human eye. The corneal segmentation technique provides valuable information for clinical diagnoses, such as corneal thickness. Non-contact anterior segment optical coherence tomography (AS-OCT) is a prevalent ophthalmic imaging technique that can visualize the anterior and posterior surfaces of the cornea. Nonetheless, during the imaging process, saturation artifacts are commonly generated due to the tangent of the corneal surface at that point, which is normal to the incident light source. This stripe-shaped saturation artifact covers the corneal surface, causing blurring of the corneal edge, reducing the accuracy of corneal segmentation. To settle this matter, an inpainting method that introduces structural similarity and frequency loss is proposed to remove the saturation artifact in AS-OCT images. Specifically, the structural similarity loss reconstructs the corneal structure and restores corneal textural details. The frequency loss combines the spatial domain with the frequency domain to ensure the overall consistency of the image in both domains. Furthermore, the performance of the proposed method in corneal segmentation tasks is evaluated, and the results indicate a significant benefit for subsequent clinical analysis.

Landscape and regulation of alternative splicing and alternative polyadenylation in a plant pathogenic fungus.

Alternative splicing (AS) and alternative polyadenylation (APA) contribute significantly to the regulation of gene expression in higher eukaryotes. Their biological impact in filamentous fungi, however, is largely unknown. Here we combine PacBio Isoform-Sequencing and strand-specific RNA-sequencing of multiple tissues and mutant characterization to reveal the landscape and regulation of AS and APA in Fusarium graminearum. We generated a transcript annotation comprising 51 617 isoforms from 17 189 genes. In total, 4997 and 11 133 genes are alternatively spliced and polyadenylated, respectively. Majority of the AS events alter coding sequences. Unexpectedly, the AS transcripts containing premature-termination codons are not sensitive to nonsense-mediated messenger RNA decay. Unlike in yeasts and animals, distal APA sites have strong signals, but proximal APA isoforms are highly expressed in F. graminearum. The 3'-end processing factors FgRNA15, FgHRP1, and FgFIP1 play roles in promoting proximal APA site usage and intron splicing. A genome-wide increase in intron inclusion and distal APA site usage and downregulation of the spliceosomal and 3'-end processing factors were observed in older and quiescent tissues, indicating intron inclusion and 3'-untranslated region lengthening as novel mechanisms in regulating aging and dormancy in fungi. This study provides new insights into the complexity and regulation of AS and APA in filamentous fungi.

High-Quality Genome Resource of Clonostachys rosea Strain CanS41 by Oxford Nanopore Long-Read Sequencing.

Clonostachys rosea is a necrotrophic mycoparasitic fungus with excellent biological control ability against numerous fungal plant pathogens. Here, we performed genomic sequencing of C. rosea strain CanS41 using Oxford Nanopore sequencing technology. We generated a high-quality genome assembly (>99.99% accuracy), which comprised 26 contigs containing 60.68 Mb sequences with a GC content of 48.55% and a repeat content of 8.38%. The N50 contig length is 3.02 Mb. In total, 20,818 protein-coding genes were identified and functionally annotated. Genes encoding secreted proteins and carbohydrate-active enzymes as well as secondary metabolic gene clusters were also identified and analyzed. In summary, the high-quality genome assembly and gene annotation provided here will allow further exploration of biological functions and enhance biological control ability of C. rosea.

Clinical Efficacy of Laminectomy with Instrumented Fixation in Treating Thoracolumbar Intradural Extramedullary Schwannomas: A Comparative Study.

BACKGROUND This study investigated the clinical effect of laminectomy plus pedicle screw fixation in treating thoracolumbar intradural extramedullary schwannomas. MATERIAL AND METHODS Between October 2011 and May 2017, 57 patients undergoing resection of thoracolumbar schwannomas were retrospectively identified and included in the study. Based on the surgical procedures used, all participants were assigned to either the laminectomy-only group (n=33) or the combination group (laminectomy plus pedicle screw fixation, n=24). All participants were followed up for over 2 years. In the laminectomy, the spinal process, vertebral laminae, and bilateral upper articular processes of the surgical segments were completely resected and the lower articular processes were reserved. For further analysis, we evaluated the pain levels using visual analogue scale (VAS) score. The assessment of neurological function was performed with Japanese Orthopaedic Association (JOA) score and Oswestry disability index (ODI). The comparisons of Cobb angle changes were carried out pre-surgery and post-surgery. RESULTS The demographic data were well matched between the laminectomy-only group and combination group, without significant differences (P>0.05). After surgery, both surgical procedures achieved significant improvement in VAS score, ODI, and JOA score (P<0.001), but no significant differences were found between these 2 surgical procedures (P>0.05). The postoperative change in Cobb angle indicated a significant difference in the laminectomy-only group, but not in the combination group (P<0.05). In addition, postoperative spinal instability/deformity was found in the laminectomy-only group (P<0.05). CONCLUSIONS In conclusion, the combination of laminectomy and pedicle screw fixation is a safe and effective surgical procedure when used to treat thoracolumbar schwannoma, and appears to be superior to the laminectomy-only procedure.

The PPAR-γ antagonist T007 inhibits RANKL-induced osteoclastogenesis and counteracts OVX-induced bone loss in mice.

BACKGROUND: Osteoclasts are key determinant cellular components implicated in the development and progression of disorders driven by bone damage. Herein, we studied the upshot of T007, an antagonist of peroxisome proliferator-activated receptor-gamma (PPARγ), on osteoclastogenesis using cell and animal models. RESULTS: The in vitro assays revealed that T007 hindered the osteoclastogenesis caused by the treatment with the receptor activator of nuclear factor-κB ligand (RANKL) through inhibiting the levels of PPARγ in cells. The PPARγ siRNA partially reproduced the inhibitory action of T007. The opposite findings were produced after PPARγ overexpression. Furthermore, T007 prevented from bone loss in a mouse model of osteoporosis induced by ovariectomy (OVX). These findings implied that T007 is a potential efficient drug for the prophylaxis and cure of osteoclast-related disorders. CONCLUSIONS: Taken together, our findings demonstrated that T007 impedes osteoclastogenesis and will be useful for the therapy of bone related diseases, essentially osteoporosis.

The methods for inserting lumbar bicortical pedicle screws from the anatomical perspective of the prevertebral great vessels.

BACKGROUND: At present, bicortical pedicle screws (BPSs) are not used clinically because they carry the potential risk of damaging the prevertebral great vessels (PGVs). The authors observed the anatomical relationship between the PGVs and simulated BPSs at different transverse screw angles (TSAs), exploring the insertion method of the BPS. METHODS: Computed tomography angiography (CTA) images from 65 adults were collected. A total of 4-5 TSAs of the BPSs were simulated on the left and right sides of L1-L5 (L1-L3: 0°, 5°, 10°, 15°; L4-L5: 0°, 5°, 10°, 15°, 20°). There were three types of distances from the anterior vertebral cortex (AVC) to the PGVs (DAVC-PGV); DAVC-PGV < 0.50 cm, DAVC-PGV ≥ 0.50 cm, and DAVC-PGV↑; these distances represented close, distant, and noncontact PGV, respectively. RESULTS: The ratio of every type of PGV was calculated, and the appropriate TSA of the BPS was recommended. In L1, the recommended left TSA of the BPS was 0°, and the ratio of the close PGV was 7.69%, while the recommended right TSA was 0°-10°, and the ratio of the close PGV was 1.54-4.62%. In L2, the recommended left TSA of the BPS was 0° and the ratio of the close PGV was 1.54%, while the recommended right TSA was 0°-15° and the ratio of the close PGV was 3.08-9.23%. In L3, the recommended left TSA was 0°-5°, and the ratio of the close PGV was 1.54-4.62%. In L4, the recommended left TSA was 0°, and the ratio of the close PGV was 4.62%. BPS use was not recommended on the right side of either L3 or L4 or on the either side of L5. CONCLUSIONS: From the anatomical perspective of the PGVs, BPSs were not suitable for insertion into every lumbar vertebra. Furthermore, the recommended methods for inserting BPSs were different in L1-L4.

A Substance Exchanger-Based Bioreactor Culture of Pig Discs for Studying the Immature Nucleus Pulposus.

Various research models have been developed to study the biology of disc cells. Recently, the adult disc nucleus pulposus (NP) has been well studied. However, the immature NP is underinvestigated due to a lack of a suitable model. This study aimed to establish an organ culture of immature porcine disc by optimizing culture conditions and using a self-developed substance exchanger-based bioreactor. Immature porcine discs were first cultured in the bioreactor for 7 days at various levels of glucose (low, medium, high), osmolarity (hypo-, iso-, hyper-) and serum (5, 10, 20%) to determine the respective optimal level. The porcine discs were then cultured under the optimized conditions in the novel bioreactor, and were compared with fresh discs at day 14. For high-glucose, iso-osmolarity, or 10% serum, cell viability, the gene expression profile (for anabolic genes and catabolic genes), and glycosaminoglycan (GAG) and hydroxyproline (HYP) contents were more favorable than for other levels of glucose, osmolarity, and serum. When the immature discs were cultured under the optimized conditions using the novel bioreactor for 14 days, the viability of the immature NP was maintained based on histology, cell viability, GAG and HYP contents, and matrix molecule expression. In conclusion, the viability of the immature NP in organ culture could be maintained under the optimized culture conditions (high-glucose, iso-osmolarity, and 10% serum) in the substance exchanger-based bioreactor.

Biological Responses of the Immature Annulus Fibrosus to Dynamic Compression in a Disc Perfusion Culture.

Mechanical stimuli participate in disc development and remodelling. However, the effects of mechanical load on the immature annulus fibrosus (AF) are largely unclear. This study aimed to investigate how the immature AF responded to dynamic compressive magnitude and duration. Immature porcine discs were bioreactor-cultured for 7 days and then dynamically compressed at various magnitudes (0.1, 0.2, 0.4, 0.8 and 1.3 MPa at a frequency of 1.0 Hz for 2 h/day) and durations (1, 2, 4 and 8 h/day at a magnitude of 0.4 MPa and a frequency of 1.0 Hz). Non-compressed discs were used as controls. The immature AF tissue was analysed for histology, gene expression (aggrecan, collagen I, ADAMTS-4, MMP-3, TIMP-1 and TIMP-3), biochemical content of glycosaminoglycans (GAG) and hydroxyproline (HYP) and aggrecan immunohistochemical staining. In the lower-compressive-magnitude groups (0.1, 0.2 and 0.4 MPa), the immature AF showed an up-regulation in the expression of matrix genes, GAG and HYP content and aggrecan deposition. In the compression duration groups, the GAG and HYP content and aggrecan deposition declined to a minimum in the 8-hour group, in which a catabolic gene expression profile was found. In conclusion, this study indicated that the effects of dynamic compression on the immature AF are magnitude and duration dependent and that catabolic remodelling within the immature AF can be induced by high compressive magnitudes and long compressive durations.

Dynamic Compression Effects on Immature Nucleus Pulposus: a Study Using a Novel Intelligent and Mechanically Active Bioreactor.

BACKGROUND: Previous cell culture and animal in vivo studies indicate the obvious effects of mechanical compression on disc cell biology. However, the effects of dynamic compression magnitude, frequency and duration on the immature nucleus pulposus (NP) from an organ-cultured disc are not well understood. OBJECTIVE: To investigate the effects of a relatively wide range of compressive magnitudes, frequencies and durations on cell apoptosis and matrix composition within the immature NP using an intelligent and mechanically active bioreactor. METHODS: Discs from the immature porcine were cultured in a mechanically active bioreactor for 7 days. The discs in various compressive magnitude groups (0.1, 0.2, 0.4, 0.8 and 1.3 MPa at a frequency of 1.0 Hz for 2 hours), frequency groups (0.1, 0.5, 1.0, 3.0 and 5.0 Hz at a magnitude of 0.4 MPa for 2 hours) and duration groups (1, 2, 4 and 8 hours at a magnitude of 0.4 MPa and frequency of 1.0 Hz) experienced dynamic compression once per day. Discs cultured without compression were used as controls. Immature NP samples were analyzed using the TUNEL assay, histological staining, glycosaminoglycan (GAG) content measurement, real-time PCR and collagen II immunohistochemical staining. RESULTS: In the 1.3 MPa, 5.0 Hz and 8 hour groups, the immature NP showed a significantly increase in apoptotic cells, a catabolic gene expression profile with down-regulated matrix molecules and up-regulated matrix degradation enzymes, and decreased GAG content and collagen II deposition. In the other compressive magnitude, frequency and duration groups, the immature NP showed a healthier status regarding NP cell apoptosis, gene expression profile and matrix production. CONCLUSION: Cell apoptosis and matrix composition within the immature NP were compressive magnitude-, frequency- and duration-dependent. The relatively high compressive magnitude or frequency and long compressive duration are not helpful for maintaining the healthy status of an immature NP.

Biomechanical comparison of anterior lumbar screw-plate fixation versus posterior lumbar pedicle screw fixation.

Anterior lumbar interbody fusion (ALIF) followed by posterior pedicle screw fixation (PSF) in a second procedure is mostly used to implement lumbar spine fusion. ALIF followed by anterior lumbar screw-plate has a lot of advantages, but its biomechanical stability requires confirmation. This study evaluated the biomechanical stability of a novel anterior lumbar locked screw-plate (ALLSP) by comparison with posterior lumbar PSF. Twelve fresh human cadaveric lumbar specimens (L4-L5) were assigned to four groups: ALIF+PSF group, ALIF+ALLSP (both fixed) group, ALIF group and an untreated control (both non-fixed) group. The first three groups received implantation of a rectangular titanium cage. Tests under axial compression, flexion, extension, lateral bending, or rotation showed that the fixed groups had significantly stronger stability than the non-fixed groups (P=0.000 for all). The ALIF+ALLSP group had significantly greater axial stiffness under applied axial compression and significantly less angular displacement under rotational forces than the ALIF+PSF group. The angular displacement of the ALIF+ALLSP group was less under flexion than that of the ALIF+PSF, and the angular displacement under lateral bending and extension was greater, but these differences were not statistically significant. In summary, the ALLSP conforms to the anterior lumbar spine and has good biomechanical stability. It is a reliable choice for enhancing the stability of ALIF.

Study on the anatomy of the lumbosacral anterior great vessels pertinent to L5/S1 anterior interbody surgery with computer tomography angiography.

We investigate the anatomy of the lumbosacral anterior great vessels using computer tomography (CT) angiography before L5/S1 anterior interbody surgery. Sixty-two adult patients were selected. The location of the abdominal aortic bifurcation and common iliac venous confluence in the lumbar vertebrae and the anatomic parameters of the iliac vascular space (e.g., distances from the included angle vertex of the iliac vascular space to the median sagittal plane and to the inferior boundary of L5 and distances between the left and right iliac vessels on the inferior boundary of L5 and on the superior boundary of S1) were analysed. Overall, 67.73% of the 62 cases had an abdominal aortic bifurcation located at L4 and L4/5 intervertebral disc; 61.29%, the common iliac venous confluence located at L5. The four distances mentioned above were 0.98 cm ± 0.38 cm, 2.01 cm ± 1.26 cm, 3.11 cm ± 1.35 cm and 4.34 cm ± 1.10 cm, respectively. A classification system of types A, B and C was developed. The calculated L5/S1 intervertebral space exposure percentages of types A, B and C were 32.21%, 82.58% and 54.68%, respectively. During L5/S1 anterior interbody surgery, type B intervertebral discs can be exposed conveniently, preventing injury of the iliac vessels, which was also observed in 54.68% and 32.21% of the type C and type A discs, respectively. Because the type A intervertebral disc has minimal exposure, the risk of iliac vascular injury is relatively high in these patients.

Comparison of Phosphogypsum-Steel Slag-Based cement and Portland cement for stabilization of heavy metals in oil-based drillings cuttings.

This research compared Portland cement and Phosphogypsum-Steel Slag-Based (PSSB) cement in terms of their capabilities to stabilize heavy metals (specifically lead and nickel) in Oil-Based Drill Cuttings (OBDC). In the experimental section, the qualitative analysis of heavy metal constituents in OBDC was captured by X-ray Photoelectron Spectroscopy (XPS). Additionally, an acetic acid leaching test was implemented for the heavy metal leaching concentration to evaluate the ceramsite stabilization effect on OBDC. In the simulation phase, cement models, heavy metal ion models, and stabilization models were constructed to explore the stabilization mechanism of heavy metals. Results demonstrated that PSSB cement exhibits superior stabilization effects on OBDC compared to Portland cement. Flame Atomic Absorption Spectrophotometry (FAAS) tests showed that PSSB cement reduced Ni and Pb leaching by 21.87 % and 47.32 %, respectively, compared to Portland cement. In PSSB cement, the diffusion coefficients for Ni and Pb ions were observed to decrease by 42.92 % and 79.63 %, respectively, as revealed through Mean Square Displacement (MSD) analysis. The cohesive energy of PSSB cement was 76.73 % lower than that of Portland cement, and its interaction energies for stabilizing Ni and Pb ions were 59.43 % and 76.22 % lower, respectively, demonstrating greater stability and efficiency in metal stabilization. PSSB cement exhibited lower heavy metal concentration and better structural stability than Portland cement.

l-arginine promotes angio-osteogenesis to enhance oxidative stress-inhibited bone formation by ameliorating mitophagy.

Background: Osteoporosis is one of the most common bone diseases in middle-aged and elderly populations worldwide. The development of new drugs to treat the disease is a key focus of research. Current treatments for osteoporosis are mainly directed at promoting osteoblasts and inhibiting osteoclasts. However, there is currently no ideal approach for osteoporosis treatment. l-arginine is a semi-essential amino acid involved in a number of cellular processes, including nitric production, protein biosynthesis, and immune responses. We previously reported that l-arginine-derived compounds can play a regulatory role in bone homeostasis. Purpose: To investigate the specific effect of l-arginine on bone homeostasis. Methods: Mildly aged and ovariectomized mouse models were used to study the effects of l-arginine on osteogenesis and angiogenesis, assessed by micro-computed tomography and immunostaining of bone tissue. The effect of l-arginine on osteogenesis, angiogenesis, and adipogenesis was further studied in vitro using osteoblasts obtained from cranial cap bone, endothelial cells, and an adipogenic cell line. Specific methods to assess these processes included lipid staining, cell migration, tube-forming, and wound-healing assays. Protein and mRNA expression was determined for select biomarkers. Results: We found that l-arginine attenuated bone loss and promoted osteogenesis and angiogenesis. l-arginine increased the activity of vascular endothelial cells, whereas it inhibited adipogenesis in vitro. In addition, we found that l-arginine altered the expression of PINK1/Parkin and Bnip3 in the mitochondria of osteoblast-lineage and endothelial cells, thereby promoting mitophagy and protecting cells from ROS. Similarly, l-arginine treatment effectively ameliorated osteoporosis in an ovariectomized mouse model. Conclusion: l-arginine promotes angio-osteogenesis, and inhibits adipogenesis, effects mediated by the PINK1/Parkin- and Bnip3-mediated mitophagy. The Translational Potential of this Article: L-arginine supplementation may be an effective adjunct therapy in the treatment of osteoporosis.

An intronic enhancer of Cebpa regulates adipocyte differentiation and adipose tissue development via long-range loop formation.

Cebpa is a master transcription factor gene for adipogenesis. However, the mechanisms of enhancer-promoter chromatin interactions controlling Cebpa transcriptional regulation during adipogenic differentiation remain largely unknown. To reveal how the three-dimensional structure of Cebpa changes during adipogenesis, we generated high-resolution chromatin interactions of Cebpa in 3T3-L1 preadipocytes and 3T3-L1 adipocytes using circularized chromosome conformation capture sequencing (4C-seq). We revealed dramatic changes in chromatin interactions and chromatin status at interaction sites during adipogenic differentiation. Based on this, we identified five active enhancers of Cebpa in 3T3-L1 adipocytes through epigenomic data and luciferase reporter assays. Next, epigenetic repression of Cebpa-L1-AD-En2 or -En3 by the dCas9-KRAB system significantly down-regulated Cebpa expression and inhibited adipocyte differentiation. Furthermore, experimental depletion of cohesin decreased the interaction intensity between Cebpa-L1-AD-En2 and the Cebpa promoter and down-regulated Cebpa expression, indicating that long-range chromatin loop formation was mediated by cohesin. Two transcription factors, RXRA and PPARG, synergistically regulate the activity of Cebpa-L1-AD-En2. To test whether Cebpa-L1-AD-En2 plays a role in adipose tissue development, we injected dCas9-KRAB-En2 lentivirus into the inguinal white adipose tissue (iWAT) of mice to suppress the activity of Cebpa-L1-AD-En2. Repression of Cebpa-L1-AD-En2 significantly decreased Cebpa expression and adipocyte size, altered iWAT transcriptome, and affected iWAT development. We identified functional enhancers regulating Cebpa expression and clarified the crucial roles of Cebpa-L1-AD-En2 and Cebpa promoter interaction in adipocyte differentiation and adipose tissue development.