Relaxin-2 promotes osteoblastic differentiation mediated by epidermal growth factor and epidermal growth factor receptor signaling.

Loss of osteogenic differentiation potential of osteoblasts has been associated with the pathogenesis of osteoporosis. Thus, stimulation of osteoblastic differentiation is a therapeutic strategy for osteoporosis. Relaxin-2 is a peptide hormone with potent biological functions. However, the effects of Relaxin-2 in osteoblastic differentiation and osteoporosis have not been reported before. Here, we report a novel physiological role of Relaxin-2 in promoting osteoblastic differentiation and mineralization of MC3T3-E1 cells. Our results indicate that exposure to Relaxin-2 upregulated the expression, and elevated the activity of alkaline phosphatase (ALP) when MC3T3-E1 cells were cultured in osteogenic differentiation medium (OM). Additionally, Relaxin-2 upregulated the mRNA levels of osteocalcin (ocn), osteopontin (opn), and collagen type I alpha 1 (Col1a1). The alizarin red S staining assay revealed that Relaxin-2 promoted the mineralization of MC3T3-E1 cells. We also found that Relaxin-2 increased the expression of Runx-2 as well as the epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR). Importantly, silencing of EGF abolished the effects of Relaxin-2 in osteoblastic differentiation and related gene expression. These findings suggest that Relaxin-2 stimulates osteogenic differentiation through activating EGF/EGFR signaling.

Surfactant-Free Synthesis of Melon Seed-Like CeO2 and Ho@CeO2 Nanostructures with Enriched Oxygen Vacancies: Characterization and Their Enhanced Antibacterial Properties.

To overcome the poor antibacterial performance of cerium oxide (CeO2) nanoparticles at low concentrations, melon seed-shaped CeO2 (MS-CeO2) and holmium (Ho)-doped CeO2 (Ho@MS-CeO2) nanoparticles were synthesized using a simple precipitation method without the addition of any surfactants. The surface morphology, phase structure, crystallinity, Ce3+ and Ce4+ valence, lattice defects, and reactive oxygen species (ROS) production of both synthesized nanostructures were examined using different techniques, i.e., scanning electron microscopy (SEM), energydispersive X-ray (EDX), resolution transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffractometry (XRD), X-ray photoelectron spectroscopy (XPS), electron paramagnetic resonance (EPR), Raman spectroscopy, ultraviolet (UV) spectra, fluorescence spectra, and zeta potential (ζ). The results show that under certain stirring and aging temperatures, CeO2 and Ho-doped CeO2 nanoparticles with a melon seed-like morphology can be prepared in a short period. Both nanoparticles were tested as antiseptic agents against G+ and G - bacteria (E. coli and S. aureus), and the results confirmed that the Ho@MS-CeO2 nanostructures exhibited remarkable antimicrobial activity at a low concentration (0.5 mg/L) compared with the control group, which is attributed to the reversible conversion of Ce3+ and Ce4+ in the ceria crystal lattice, enriched oxygen vacancy, ROS species production, and positive surface charge.

Electrokinetic motion and viability assessment of algae with a polyethylene glycol-dextran interface.

At present, there is still limited report on the electrokinetic (EK) behavior of bioparticles at the interface of an aqueous two-phase system. In this paper, the EK motion and viability assessment of live algae mixed with the NaClO treated dead algae were carried out at the interface formed by polyethylene glycol (PEG)-rich phase and dextran (DEX)-rich phase in a straight microchannel. The experimental results show that both the live and dead algae at the PEG-DEX interface migrate from the negative electrode to the positive electrode, and the EK velocity of live algae at the interface is slightly larger than that of the dead ones with similar diameters. For either live or dead algae, the EK velocity at the interface decreases with the increase in diameter. A size-velocity curve was used to evaluate the viability of the algae. As most of the microorganisms in ballast water are algae, the method in this paper provides a promising way to detect and evaluate the live microorganism in treated ballast water of a ship.

[Clinical application of neurovascular staghorn flap for repairing of defects in fingertips].

Objective: To evaluate the effectiveness of neurovascular staghorn flap for repairing defects in fingertips. Methods: Between August 2019 and October 2021, a total of 15 fingertips defects were repaired with neurovascular staghorn flap. There were 8 males and 7 females with an average age of 44 years (range, 28-65 years). The causes of injury included 8 cases of machine crush injury, 4 cases of heavy object crush injury, and 3 cases of cutting injury. There were 1 case of thumb, 5 cases of index finger, 6 cases of middle finger, 2 cases of ring finger, and 1 case of little finger. There were 12 cases in emergency, and 3 cases with finger tip necrosis after trauma suture. Bone and tendon exposed in all cases. The range of fingertip defect was 1.2 cm×0.8 cm to 1.8 cm×1.5 cm, and the range of skin flap was 2.0 cm×1.5 cm to 2.5 cm×2.0 cm. The donor site was sutured directly. Results: All flaps survived without infection or necrosis, and the incisions healed by first intention. All patients were followed up 6-12 months, with an average of 10 months. At last follow-up, the appearance of the flap was satisfactory, the wear resistance was good, the color was similar to the skin of the finger pulp, and there was no swelling; the two-point discrimination of the flap was 3-5 mm. One patient had linear scar contracture on the palmar side with slight limitation of flexion and extension, which had little effect on the function; the other patients had no obvious scar contracture, good flexion and extension of the fingers, and no dysfunction. The finger function was evaluated according to the total range of motion (TAM) system of the Hand Surgery Society of Chinese Medical Association, and excellent results were obtained in 13 cases and good results in 2 cases. Conclusion: The neurovascular staghorn flap is a simple and reliable method to repair fingertip defect. The flap has a good fit with the wound without wasting skin. The appearance and function of the finger are satisfactory after operation.

LncRNA SNHG5 promotes chondrocyte proliferation and inhibits apoptosis in osteoarthritis by regulating miR-10a-5p/H3F3B axis.

BACKGROUND: Osteoarthritis (OA) is a common degenerative joint disease in the elderly. Increasing evidence suggested that long non-coding RNAs (lncRNAs) played vital roles in OA progression. This study aimed to explore the role and mechanism of lncRNA small nucleolar RNA host gene 5 (SNHG5) in OA development. METHODS: Chondrocytes were stimulated with interleukin-1ß (IL-1ß) in vitro. The levels of SNHG5, miR-10a-5p, and H3 histone family 3B (H3F3B) were measured by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot. Cell proliferation was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and colony formation assay. Cell apoptosis was tested by flow cytometry. The levels of apoptosis-related and cartilage-related markers were detected by western blot. The interaction among SNHG5, miR-10a-5p, and H3F3B was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. RESULTS: SNHG5 and H3F3B were downregulated, while miR-10a-5p was upregulated in OA cartilage tissues. Knockdown of SNHG5 enhanced IL-1ß-induced apoptosis in chondrocytes. Rescue experiments verified that SNHG5 hindered apoptosis in IL-1ß-stimulated chondrocytes by sponging miR-10a-5p. Moreover, H3F3B was a target of miR-10a-5p, and miR-10a-5p promoted IL-1ß-induced chondrocyte apoptosis by regulating H3F3B. In addition, SNHG5 regulated H3F3B expression via sponging miR-10a-5p in IL-1ß-treated chondrocytes. CONCLUSION: SNHG5 suppressed chondrocytes apoptosis in OA by regulating the miR-10a-5p/H3F3B axis, which provided a promising biomarker for OA treatment.

Effect and Mechanism of the Bruton Tyrosine Kinase (Btk) Inhibitor Ibrutinib on Rat Model of Diabetic Foot Ulcers.

BACKGROUND Diabetes causes damage to the soft tissue and bone structure of the foot, referred to as "diabetic foot". Ibrutinib is a Bruton tyrosine kinase (Btk) inhibitor, and the role and mechanism of ibrutinib on the diabetic foot have not been elucidated. MATERIAL AND METHODS Male Wister rats were randomly divided into 3 groups: control group, model group, and ibrutinib group. After 14 days, the ulcer wound size of each group was measured, and the ulcer healing rate was calculated. The level of inflammatory factors interleukin (IL)-1ß, tumor necrosis factor (TNF)-alpha, and IL-6 was detected by enzyme-linked immunosorbent assay (ELISA). Real-time polymerase chain reaction (PCR) was used to analyze the changes of Toll-like receptor 2 (TLR2) and TLR4. The expression of vascular endothelial growth factor (VEGF) and the RAGE (receptor for advanced glycation end product/NF-kappaB (nuclear factor-kappa B) pathway was detected by western blot. RESULTS Blood glucose, blood lipids, serum creatinine, and urea nitrogen (BUN) levels were increased in the model group, together with increased levels of IL-1ß, TNF-alpha, IL-6, as well as TLR2 and TLR4 expression, and there were significant differences compared with the control group (P<0.05). Meanwhile, the model group showed decreased VEGF expression and increased expression of RAGE and NF-kappaB. However, ibrutinib reduced blood sugar, blood lipids, creatinine, and urea nitrogen levels, inhibited the secretion of inflammatory factors, promoted ulcer healing, improved ulcer healing rate, decreased the expression of TLR2, TLR4, RAGE, and NF-kappaB, and increased VEGF expression; there were significant differences in the ibrutinib group compared with the model group (P<0.05). CONCLUSIONS The Btk inhibitor ibrutinib can upregulate VEGF expression, inhibit the expression of TLRs, inhibit the secretion of inflammatory factors, and promote the healing of diabetic foot ulcer possibly by regulating the RAGE/NF-kappaB pathway.

Polymeric hole-shaped polyhedral aggregates: Preparation, characterization, and antibacterial adhesion properties.

The diverse morphologies of aggregates formed by the self-assembly of block copolymers in selective solvents have attracted widespread attention, but the design of aggregate shapes is still limited owing to the thermodynamic favorability of sphere formation. In this paper, we report our discovery that polyhedral aggregates can be formed by the self-assembly of 1H,1H,2H,2H-perfluoro-1-dodecanol (PFD)-grafted amphiphilic polystyrene-b-poly(acrylic acid) (PS-b-PAA-g-PFD) copolymers in water at room temperature. It is determined that the crystallization of fluorocarbon side chains at the surface of PS-b-PAA-g-PFD aggregates induces the formation of a polyhedral shape. The morphology of aggregates can be controlled by the dialysis temperature, the grafting ratio of PFD in PS-b-PAA-g-PFD copolymers, and the initial copolymer concentration. The layers of polyhedral aggregates show excellent antibacterial adhesion properties. We anticipate that this method will expand the promise of self-assembly for the synthesis of a series of nonspherical micellar nanoparticles, which have promising applications in various fields.

Gene Modification of Transforming Growth Factor ß (TGF-ß) and Interleukin 10 (IL-10) in Suppressing Mt Sonicate Induced Osteoclast Formation and Bone Absorption.

BACKGROUND Osteoarticular tuberculosis is an osteolytic lesion caused by Mycobacterium tuberculosis (MTB). Inflammatory factors such as TNF-α play a critical role in anti-tuberculosis immunity by regulating osteoblast and osteoclast functions. Both TGF-ß and IL-10 have immune suppression effects to downregulate secretion and release of inflammatory factors, such as TNF-α, that play roles in regulating osteoblast and osteoclast functions. This study thus investigated the effects of osteoclast with modified TGF-ß and IL-10 gene expression on MTB-induced osteoclast formation and bone absorption. MATERIAL AND METHODS Bone marrow mononuclear cells were induced to differentiate into osteoblasts and osteoclasts in vitro to generate a co-culture system. MTB powder lysed by ultrasound (Mt sonicate) were added in gradients to observe osteoblast formation and osteoclast absorption. Cell apoptosis was measured by flow cytometry, while ELISA was used assess TNF-α, TGF-ß, and IL-10. Viral vectors carrying TGF-ß or IL-10 gene were used to transfect osteoclasts, followed by ELISA assay. Bone absorption and osteoblast apoptosis were compared among groups. RESULTS Mt sonicate significantly facilitated osteoclast formation and bone formation. It upregulated contents of TNF-α, TGF-ß, and IL-10, induced osteoblast apoptosis, enhanced RANKL expression in osteoblasts, and decreased OPG expression. Overexpression of TGF-ß and/or IL-10 significantly decreased its upregulation effect on TNF-α by Mt sonicate, and hindered Mt sonicate-induced osteoblast apoptosis, osteoclast formation, and bone absorption. CONCLUSIONS Overexpression of TGF-ß and IL-10 significantly inhibits TMB-induced TNF-α synthesis and release, suppresses osteoblast apoptosis, and hinders osteoclast formation and bone absorption.

Autofluorescent Polymers: 1 H,1 H,2 H,2 H-Perfluoro-1-decanol Grafted Poly(styrene- b-acrylic acid) Block Copolymers without Conventional Fluorophore.

Recently, although several unconventional luminescent polymers have been synthesized, it still remains a significant challenge to prepare various new fluorescent polymers by functionalization of nonfluorescent polymers. A nonfluorescent 1 H,1 H,2 H,2 H-perfluoro-1-decanol grafted to nonfluorescent polystyrene- b-poly(acrylic acid) block copolymers through simply esterification reaction can exhibit strong blue emission. On the basis of control experiments and theoretical simulation, we have proposed that the luminescence stems from interchain n → π* interaction between the lone pair (n) of hydroxyl O atoms of carboxyl units and empty π* orbital of ester carbonyl unit. In addition, the fluorescent polymers are successfully employed for fluorescence imaging in living HeLa cell.

[Expression of Wnt5a in dorsal root ganglia in rat model of chronic compression of dorsal root ganglia].

OBJECTIVE: To investigate the role of Wnt5a in the mechanism of radiculopathy and the relation between Wnt5a and tumor necrosis factor α (TNF-α) by observing the change of the expression of Wnt5a in the rat model of chronic compression of dorsal root ganglia (CCD). METHODS: A total of 192 adult male Sprague Dawley rats were allocated into 4 groups: shame group (group A, n = 48), CCD group (group B, n = 48), CCD + saline group (group C, n = 48), and CCD + etanercept group (group D, n = 48). An L-shaped needle (about 3.5 mm in length, 0.6 mm in diameter) was inserted into the L5 intervertebral foramen, and the dorsal root ganglia (DRG) was compressed by the needle to prepare the CCD model in groups B, C, and D, and then normal saline (5.5 mg/kg) or etanercept was injected intraperitoneally in groups C and D. The intervertebral foramen was exposed in group A. The mechanical pain threshold of the posterior paw was tested by the von Frey filaments at 1, 3, 5, and 7 days after operation; the expressions of Wnt5a protein and mRNA were detected at 3 and 7 days after operation by immunohistochemical staining and RT-PCR, respectively. RESULTS: The mechanical pain threshold of groups B and C was significantly lower than that of groups A and D, and in group D than in group A (P < 0.05), but no significant difference was found between groups B and C (P > 0.05). The Wnt5a positive cells and the mRNA expression of Wnt5a at 7 days were significantly more than those at 3 days in groups B, C, and D (P < 0.05). The Wnt5a positive cells and the mRNA expression of Wnt5a in groups B and C were significantly more than in groups A and D, and in group D than in group A (P < 0.05), but no significant difference was shown between groups B and C (P > 0.05). CONCLUSION: The expression of Wnt5a in the DRG is increased after CCD. The expression of Wnt5a in the DRG is decreased after the administration of the inhibitor of TNF-α.