Genome editing in plants using the TnpB transposase system.

The widely used clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease (Cas) system is thought to have evolved from IS200/IS605 transposons. TnpB proteins, encoded by one type of IS200/IS605 transposon, are considered to be the evolutionary ancestors of Cas12 nucleases, which have been engineered to function as RNA-guided DNA endonucleases for genome editing in bacteria and human cells. TnpB nucleases, which are smaller than Cas nucleases, have been engineered for use in genome editing in animal systems, but the feasibility of this approach in plants remained unknown. Here, we obtained stably transformed genome-edited mutants in rice (Oryza sativa) by adapting three recently identified TnpB genome editing vectors, encoding distinct TnpB nucleases (ISAam1, ISDra2, and ISYmu1), for use in plants, demonstrating that the hypercompact TnpB proteins can effectively edit plant genomes. ISDra2 and ISYmu1 precisely edited their target sequences, with no off-target mutations detected, showing that TnpB transposon nucleases are suitable for development into a new genome editing tool for plants. Future modifications improving the genome-editing efficiency of the TnpB system will facilitate plant functional studies and breeding programs. Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-024-00172-6.

Unveiling the Therapeutic Potential of Herba Epimedii: Enhancing Bone Healing Through Cytoskeletal Regulation of RhoA/Rock1 Pathway.

Herba Epimedii is widely used to promote bone healing, and their active ingredients are total flavonoids of Epimedium (TFE). Ras homolog gene family member A / Rho-associated protein kinase (RhoA/Rock), an important pathway regulating the cytoskeleton, has been proven to affect bone formation. However, whether TFE promotes bone healing via this pathway remains unclear. In this study, the therapeutic effects of TFE were estimated using micro-computed tomography and hematoxylin and eosin staining of pathological sections. F-actin in osteoblasts was stained to investigate the protective effects of TFE on the cytoskeleton. Its regulatory effects on the RhoA/Rock1 pathway were explored using RT-qPCR and Western blot analysis. Besides, flow cytometry, alkaline phosphatase and nodule calcification staining were performed to evaluate the effects on osteogenesis. The bone healing in rats was improved, the cytoskeletal damage in osteoblasts was reduced, the RhoA/Rock1 pathway was downregulated, and osteogenesis was enhanced after TFE treatment. Thus, TFE can promote bone formation at least partially by regulating the expression of key genes and proteins in the cytoskeleton. The findings of this study provided evidence for clinical applications and would contribute to a better understanding of Epimedium's mechanisms in treating bone defects.

PbTeB4O9: a lead tellurium borate with unprecedented fundamental building block [B4O10] and large birefringence.

Herein, the first lead tellurium borate, PbTeB4O9, with an unprecedented fundamental building block [B4O10] was successfully synthesized. The near-parallel alignment of [B4O10] groups and [TeO3] polyhedra resulted in a high birefringence (0.099@1064 nm). The structure-property relationship was discussed by using the first-principles calculations.

The effects of mulberry polyphenols on the digestibility and absorption properties of pork myofibrillar protein in vitro.

The objective of the present study was to explore the effect of mulberry polyphenols on the digestibility and absorption properties of myofibrillar protein (MP) in vitro. MP was extracted from the Longissimus et thoracis muscle of 18 different pig carcasses and the MP-mulberry polyphenols complex was prepared. The antioxidant activity of digestive juice, degradation of both MP and polyphenols, and the metabolism of MP and the MP-polyphenols complex by intestinal microbial activity during digestion and fermentation in vitro were compared. The results showed that mulberry polyphenols significantly affect the digestibility of MP and the antioxidant activity of digestive juices during digestion (P < 0.05). After the modification of the polyphenols, the hydrolysis of MP increased from 55.4% to 64.0%, and the molecular weight of protein digestion product significantly decreased (P < 0.05). The scavenging rates of 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl in the final digestive juice were 350.1 µmol Trolox/mg protein and 34.0%, respectively, which were 0.34 and 0.47-fold higher than those of the control (P < 0.05). Furthermore, the release and degradation of phenolic compounds mainly occurred during intestinal digestion, and polyphenols that reached the colon after digestion, through the fermentation of intestinal microorganisms in vitro, enriched Lactobacillus and promoted the production of short-chain fatty acids which has obvious potential to improve intestinal health.

Na2BaB12O18F4: A Mixed Alkali/Alkaline-Earth Metal Fluorooxoborate with Two Unprecedented Interpenetrating Three-Dimensional B-O/F Anionic Networks and a Short Ultraviolet Cutoff Edge.

Fluorooxoborates are promising yet largely untapped crystal materials for linear and nonlinear optical applications. The introduction of a strong electronegative F atom into an oxyboron anionic group offers a virtually unlimited chance for structural engineering and ultimately purposeful tuning of the macroscopic optical properties of the crystal. Herein, a new mixed alkali/alkaline-earth fluorooxoborate, Na2BaB12O18F4, was synthesized in a closed system. Na2BaB12O18F4 features a [B6O11F2] fundamental building unit (FBB), which polymerizes into two new (first example) independent interpenetrating three-dimensional (3D) B-O/F anionic networks constructed entirely from BO3 and BO3F units. Based on optical characterizations and the first-principles calculations, Na2BaB12O18F4 exhibits a moderate birefringence (0.054 @ 1064 nm) and a short ultraviolet (UV) cutoff edge (below 190 nm). The successful synthesis and characterization of Na2BaB12O18F4 may speed up the subsequent discovery of other mixed alkali/alkaline-earth metal fluorooxoborates.

Wide-Temperature Tunable Phonon Thermal Switch Based on Ferroelectric Domain Walls of Tetragonal KTN Single Crystal.

Ferroelectric domain walls (DWs) of perovskite oxide materials, which can be written and erased by an external electric field, offer the possibility to dynamically manipulate phonon scattering and thermal flux behavior. Different from previous ferroelectric materials, such as BaTiO3, PbTiO3, etc., with an immutable and low Curie temperature. The Curie temperature of perovskite oxide KTa1-xNbxO3 (KTN) crystal can be tuned by altering the Ta/Nb ratio. In this work, the ferroelectric KTa0.6Nb0.4O3 (KTN) single crystal is obtained by the Czochralski method. To understand the role of ferroelectric domains in thermal transport behavior, we perform a nonequilibrium molecular dynamics (NEMD) calculation on monodomain and 90° DWs of KTN at room temperature. The calculated thermal conductivity of monodomain KTN is 9.84 W/(m·k), consistent with experimental results of 8.96 W/(m·k), and distinctly decreased with the number of DWs indicating the outstanding performance of the thermal switch. We further evaluate the thermal boundary resistance (TBR) of KTN DWs. An interfacial thermal resistance value of 2.29 × 10-9 K·m2/W and a large thermal switch ratio of 4.76 was obtained for a single DW of KTN. Our study shows that the ferroelectric KTN can provide great potential for the application of thermal switch at room temperature and over a broad temperature range.

Improved physicochemical stability and bioaccessibility of astaxanthin-loaded oil-in-water emulsions by a casein-caffeic acid-glucose ternary conjugate.

In this study, the influence of casein-caffeic acid-glucose ternary conjugate (CSC) on the physicochemical properties and bioaccessibility of astaxanthin-loaded emulsion was investigated and compared with sodium caseinate (CSN), a synthetic emulsifier commonly used in the food industry. The CSC-stabilized emulsion exhibits droplet characteristics similar to CSN-stabilized emulsion, and can effectively resist the external forces that lead to the phase separation of the emulsion. Although phase separation also occurred at pH 4.0, CSC emulsion had a wider range of pH stability (pH 3.0, 5.0-8.0) and higher salt ion stability than CSN emulsion. Furthermore, CSC-stabilized astaxanthin emulsions showed better astaxanthin protection under different heat treatment conditions and storage temperatures compared with CSN. After 28 days of storage at 4 °C, astaxanthin residues in the CSC-stabilized emulsion reached 92.37 %. The bioaccessibility of astaxanthin in CSC-stabilized emulsion was 26.21 %, much higher than that in CSN (6.47 %). This research study provides a platform for designing astaxanthin-fortified food or beverage systems to achieve better stability and delivery to target sites.

Enhancement of water barrier and antimicrobial properties of chitosan/gelatin films by hydrophobic deep eutectic solvent.

Biodegradable chitosan/gelatin (CS/GEL) films have attracted attention as food packaging, but the poor water sensitivity and functional limitations of these films should be addressed. In this study, the hydrophobic deep eutectic solvent (DES, 0-15 %) consisting of thymol and octanoic acid was used to improve the water resistance and antibacterial performance of the CS/GEL composite films. FTIR and SEM analyses revealed a strong interaction between the CS/GEL matrix and DES. The films blended with DES showed increased water contact angle values and thermal stability. Furthermore, the addition of DES resulted in a significant increase in the elasticity and decrease water vapor transmission rate (WVTR). The CS/GEL films blended with 9% DES showed a 38.5% decrease in WVTR compared to those without DES. Additionally, the DES-containing film displayed good antibacterial activity against Staphylococcus aureus and Escherichia coli. Overall, the CS/GEL-DES composite films are expected to contribute an improvement to food packaging.

Hybrid Machine Learning Approach for Evapotranspiration Estimation of Fruit Tree in Agricultural Cyber-Physical Systems.

The flourish of the Internet of Things (IoT) and data-driven techniques provide new ideas for enhancing agricultural production, where evapotranspiration estimation is a crucial issue in crop irrigation systems. However, tremendous and unsynchronized data from agricultural cyber-physical systems bring large computational costs as well as complicate performing conventional machine learning methods. To precisely estimate evapotranspiration with acceptable computational costs under the background of IoT, we combine time granulation computing techniques and gradient boosting decision tree (GBDT) with Bayesian optimization (BO) to propose a hybrid machine learning approach. In the combination, a fuzzy granulation method and a time calibration technique are introduced to break voluminous and unsynchronized data into small-scale and synchronized granules with high representativeness. Subsequently, GBDT is implemented to predict evapotranspiration, and BO is utilized to find the optimal hyperparameter values from the reduced granules. IoT data from Xi'an Fruit Technology Promotion Center in Shaanxi Province, China, verify that the proposed granular-GBDT-BO is effective for cherry tree evapotranspiration estimation with reduced computational time, and acceptable and robust predictive accuracy. Consequently, the precise estimation of crop evapotranspiration could provide operational guidance for plant irrigation, plant conservations, and pest control in the agricultural greenhouse.

Influence of Gelatin-Chitosan-Glycerol Edible Coating Incorporated with Chlorogenic Acid, Gallic Acid, and Resveratrol on the Preservation of Fresh Beef.

Chlorogenic acid (CA), gallic acid (GA), and resveratrol (RES) were added to a gelatin (GEL)-chitosan (CHI)-glycerol (GLY) edible coating, and their effects on the coating of fresh beef preservation were investigated. The results revealed that CA had the most significant improvement effect on fresh beef preservation. The combination of GEL-CHI-GLY-CA preserved the color of the beef better and delayed the increase of the total volatile base nitrogen, even though its total phenolic content decreased at a faster rate during beef preservation. GA also improved the preservation effect as on the 12th day of storage, the beef samples treated with GEL-CHI-GLY-GA had the lowest thiobarbituric acid reactive substances (0.76 mg Malondialdehyde (MDA)/kg) and total viable count (6.0 log cfu/g). On the whole, though RES showed an improvement on beef preservation, the improvement was not as good as the other two polyphenols. After 12 days of storage, the beef samples treated with GEL-CHI-GLY-RES had a higher pH value (6.25) than the other two polyphenol treatmed groups. Overall, the three polyphenol-added combinations increased the shelf life of beef by approximately 3-6 days compared to the control group (treated GEL-CHI-GLY with distilled water).

Femtosecond Laser Induced Lattice Deformation in KTN Crystal.

In recent years, many novel optical phenomena have been discovered based on perovskite materials, but the practical applications are limited because of the difficulties of device fabrication. Here, we propose a method to directly induce localized lattice modification inside the potassium tantalate niobate crystal by using the femtosecond laser. This selective modification at the processed regions and the surrounding areas is characterized by two-dimensional Raman spectrum mapping. The spectrum variations corresponding to specific lattice vibration modes demonstrate the lattice structure deformation. In this way, the lattice expansion at the femtosecond laser irradiated regions and the lattice compression at the surrounding areas are revealed. Furthermore, surface morphology measurement confirms this lattice expansion and suggests the extension of lattice structure along the space diagonal direction. Moreover, the existence of an amorphization core is revealed. These modifications on the sample lattice can induce localized changes in physicochemical properties; therefore, this method can realize the fabrication of both linear diffraction and nonlinear frequency conversion devices by utilizing the novel optical responses of perovskite materials.

Ho2O3-TiO2 Nanobelts Electrode for Highly Selective and Sensitive Detection of Cancer miRNAs.

The design and engineering of effective electrode materials is critical in the development of electrochemical sensors. In the present study, Ho2O3-TiO2 nanobelts were synthesized by an alkaline hydrothermal process. The structure and morphology were investigated by X-ray diffraction (XRD) and field emission scanning electron microscope (FESEM) measurements. The Ho2O3-TiO2 nanobelts showed a distinctly enhanced (004) reflection peak and rough surfaces and were used for the electrochemical selective sensing of various cancer miRNAs, such as prostate cancer miR-141, osteosarcoma miR-21, and pancreatic cancer miR-1290. Voltammetric measurements showed an oxidation peak at +0.4, +0.2, and +1.53 V for the three different cancer biomarkers, respectively, with the detection limit as low as 4.26 aM. The results suggest that the Ho2O3-TiO2 nanobelts can be used as active materials to detect early cancers, for in vitro screening of anticancer drugs, and molecular biology research.

The miRNA-144-5p/IRS1/AKT axis regulates the migration, proliferation, and mineralization of osteoblasts: A mechanism of bone repair in diabetic osteoporosis.

Diabetic osteoporosis (DOP) is a disorder of bone metabolism induced by multiple mechanisms. Previous studies have revealed that microRNAs (miRNAs) play crucial roles in bone metabolism. MiRNA-144-5p has been proven to participate in the regulation of osteoblast activities; however, its specific mechanism in DOP has not been elucidated. This study investigated whether high glucose (HG) inhibited osteoblasts by regulating miRNA-144-5p. Our results showed that HG inhibited bone formation not only in vivo but also in vitro. We observed that HG severely hindered the migration, proliferation and mineralization of osteoblasts, while miRNA-144-5p was upregulated by way of the cell counting kit-8 assay, wound healing assay, alkaline phosphatase (ALP) activity assay and alizarin red staining. Double luciferase reporter experiments showed that miRNA-144-5p directly targeted insulin receptor substrate 1 (IRS1). The IRS1/AKT signaling pathway is closely related to osteoblasts' migration, proliferation, and mineralization. Silencing miRNA-144-5p promoted the mRNA, and protein expression of IRS1, thereby letting the expression of total AKT down, and then preventing phosphorylation of AKT into the nucleus to regulate migration, proliferation, and mineralization genes of osteoblasts. In conclusion, this study indicated that HG regulated the migration, proliferation, and mineralization of osteoblasts via the miRNA-144-5p/IRS1/AKT axis, which suggested a possible mechanism for DOP pathology.

Influences of miR-378a-3p on the Pathogenesis of Allergic Rhinitis via GzmB-Mediated Inflammatory Reaction.

Methods: Totally, 24 BALB/c mice were assigned to the AR group, control group, GzmB group, and blank group (each n = 6). The blank group was normally fed without treatment, and the other three groups were treated by ovalbumin (OVA) to induce AR models, in which the GzmB group was intranasally injected with lentiviral vector suppressing GzmB expression during the second immunization, while the control group was given the GzmB-blank vector. The times of AR pathological behaviours such as sneezing and scratching the nose of mice were observed and counted. The nasal lavage fluid of each mouse was acquired, and then, the mouse was executed by cervical dislocation, followed by collection of blood and nasal mucosa tissues. Then, ELISA was adopted for quantifying immunoglobulin E (IgE), interleukin (IL)-4, IL-6, and histamine (HA), and nasal mucosa tissues were treated by HE and TUNEL staining to observing their histopathological manifestations. PCR and western blot (WB) were adopted for quantifying GzmB and miR-378a-3p. Additionally, with NP69 cells, dual luciferase reporter (DLR) assay was carried out for determining the targeting association of GzmB with miR-378a-3p. Another 24 mice were assigned to the AR group, GzmB group, miR-378a-3p group, and GzmB+ miR-378a-3p group (each n = 6). The AR and GzmB groups were treated as above. The miR-378a-3p group was intervened by lentiviral vector suppressing miR-378a-3p, while the GzmB+ miR-378a-3p group was given GzmB and lentiviral vector suppressing miR-378a-3p meantime. A rescue assay was conducted through repeating the above tests. Results: The times of sneezing and rubbing the nose and the levels of IgE, IL-4, IL-6, and HA were similar between the control and AR groups (all P > 0.05), and these items of the two groups were all higher than those of the blank and GzmB groups (all P < 0.05). However, no notable difference was observed in IL-4 and IL-6 levels between the GzmB and blank groups (both P > 0.05), while higher levels of other detection results were found in the former group than in the latter (all P < 0.05). The staining results revealed obvious congestion, oedema, and necrosis structures in the nasal mucosa epithelium of the control and AR groups and also revealed a large number of infiltrating eosinophils and notable increase of apoptotic nasal mucosa epithelial cells. The GzmB group showed notably improved nasal mucosa tissues, and its infiltration and apoptosis of eosinophils were more notable than those of the blank group, but notably weaker than those of the AR and control groups. Additionally, the PCR and WB results revealed similar miR-378a-3p and GzmB levels in nasal mucosa between the control and AR groups (both P > 0.05), and a notable decrease of miR-378a-3p and a notable increase of GzmB in both groups (both P < 0.05). The DLR assay revealed notably suppressed fluorescence activity of GzmB-WT in NP69 cells after transfection of miR-378a-3p mimics (P < 0.05) and notably down regulated GzmB protein after increase of miR-378a-3p (P<0.05). Finally, the rescue assay revealed that downregulating miR-378a-3p aggravated the pathological changes of AR (P < 0.05) and also completely reversed the impacts of inhibiting GzmB on the pathological behaviours of AR mice. Conclusions: MiR-378a-3p can accelerate the pathological development of AR through targeted inhibition on the release of pro-inflammatory factors such as IgE and HA activated by GzmB, so it is a promising molecular target of AR therapy and offers a novel research direction for the complete cure of AR.

Efficient CRISPR/Cas9 system based on autonomously replicating plasmid with an AMA1 sequence and precisely targeted gene deletion in the edible fungus, Cordyceps militaris.

Cordyceps militaris is a popular edible fungus with important economic value worldwide. In this study, an efficient CRISPR/Cas9 genome-editing system based on an autonomously replicating plasmid with an AMA1 sequence was constructed. Further, a precisely targeted gene deletion via homology-directed repair was effectively introduced in C. militaris. Gene editing was successful, with efficiencies of 55.1% and 89% for Cmwc-1 and Cmvvd, respectively. Precisely targeted gene deletion was achieved at an efficiency of 73.9% by a single guide RNA supplementation with donor DNAs. Double genes, Cmwc-1 and Cmvvd, were edited simultaneously with an efficiency of 10%. Plasmid loss was observed under non-selective culture conditions, which could permit recycling of the selectable marker and avoid the adverse effects of the CRISPR/Cas9 system on the fungus, which is beneficial for the generation of new cultivars. RNA Pol III promoters, endogenous tRNAPro of C. militaris, and chimeric AfU6-tRNAGly can be used to improve the efficiency. Polyethylene glycol-mediated protoplast transformation was markedly more efficient than Agrobacterium tumefaciens-mediated transformation of C. militaris. To our knowledge, this is the first description of genome editing and precisely targeted gene deletion in mushrooms based on AMA1 plasmids. Our findings will enable the modification of multiple genes in both functional genomics research and strain breeding.

Enormous electro-optic effect in paraelectric nanodisordered KTa1-xNbxO3 crystal.

Recent experiments have revealed that the order of the electro-optic (EO) effect depends on the frequency of electric field in paraelectric nanodisordered KTa1-xNbxO3 (KTN) crystal. Through the dielectric frequency spectrum under the bias electric field, enormous linear and quadratic EO effects were discovered at the resonance frequencies, which changed the perception that only the quadratic EO effect exists. Applying just a small AC electric field of 6 V/mm, the effective linear EO coefficient reached 478 pm/V at 609 kHz, and the effective quadratic EO coefficient reached 4.39*10-13m2V-2at 302 kHz. The reason why an extremely low electric field results in an enormous EO coefficient is attributed to the resonance between the polar nanoregions (PNRs) and the electric field, induced by the field-driven reorientation of free dipoles on the boundary of the PNRs. In addition, the order of EO effect depending on the frequency of electric field was attributed to the motion modes of the PNR. This finding improves the understanding of how the EO effect is caused by field-driven PNR dynamics, but also provides a basis for the development of EO devices.

Variable dimensionality of the anion framework in four new borophosphates and fluoroborophosphates with short cutoff edges.

Two new borophosphates, Cs3B3P4O16 and Li(NH4)2B3P4O16, and two new fluoroborophosphates, K4B2P2O9F2 and Rb3B2PO5F4, have been obtained via the high-temperature solution method. Single-crystal X-ray diffraction indicates that all of them exhibit various anion frameworks although they crystallize in the same space group, P21/c. Two-dimensional (2D) [B3P4O16]∞ layers and a 3D [B3P4O16]∞ network can be found in Cs3B3P4O16 and Li(NH4)2B3P4O16, respectively, while isolated [B2P2O9F2] and [B2PO5F4] exist in K4B2P2O9F2 and Rb3B2PO5F4, respectively. The effect of the alkali metal cation size on the framework structures of Cs3B3P4O16 and Li(NH4)2B3P4O16 has been discussed in detail. The IR spectra confirm their structural validity. UV-Vis-NIR diffuse reflectance spectroscopy indicates that the new compounds exhibit short cutoff edges. In addition, theoretical calculations were carried out to understand their electronic structures and optical properties.

Inverse Design of Ferroelectric-Order in Perovskite Crystal for Self-Powered Ultraviolet Photodetection.

It has always been a hot topic to design an orderly mesoscopic structure in functional materials to tailor the macroscopic properties or realize new functions. The existence of domains in ferroelectric materials has been proven to affect the macroscopic properties, being actively studied in nonlinear optical conversion and piezoelectric effects. However, the high-efficiency photoelectric conversion capability of ferroelectric crystals has not yet been explored. Here, the authors study the orderly arrangement of ferroelectric order in KTa1- x Nbx O3 (KTN) perovskite crystals, and design the "head-to-head" domains by tuning the Curie temperature Tc , thereby generating abundant charged domain walls and robust conductive channels for electrons and holes. An ultrahigh ultraviolet photoresponsivity is achieved in the KTN crystal under zero bias voltage, being about four orders magnitude higher than that of the well-known ferroelectric materials. The substantial improvement can be attributed to the judiciously designed ferroelectric order, as demonstrated by the conductive atomic force microscopy. In addition, KTN detector exhibits high stability and reliability after high-temperature and fatigue treatment. KTN crystal features giant photoresponsivity, high electric-optical coefficient, and large χ(2) nonlinearity concurrently, indicating its great potential for application of all-optical devices on photonic chips.

Erxian herbal pair enhances bone formation in infected bone nonunion models and attenuates lipopolysaccharide-induced osteoblastinhibition by regulating miRNA-34a-5p.

Bacterium-induced inflammatory responses cause bone nonunion. Although antibiotics suppress infection, bone loss after antibacterial treatment remains a critical challenge. Erxian herbal pair (EHP) has been proven effective in promoting bone formation. Our study aimed to investigate the effect of EHP on bone repair after anti-infection treatment, explore its effect on a lipopolysaccharide (LPS)-induced osteoblast. We evaluated effects of EHP on bone repair with Micro-CT, and morphology detecting. Chemical constituents of EHP and EHP-containing serum (EHP-CS) were identified by UHPLC-Q/TOF-MS. In addition, osteoblast induced by LPS was established and administrated with EHP-CS. Cell proliferationwas assessed by MTT. Target prediction identified SMAD2 as a potential target of miRNA-34a-5p. MiRNA mimic, inhibitor and siRNA were transiently transfected into osteoblasts. The mRNA levels and protein expressions of miRNA-34a-5p, BMP2, Runx2, SMAD2 were assessed. The results showed that the main biocactivity ingredients in EHP-CS were Baohuoside Ι and Orcinol Glucoside. EHP could promote bone remolding after anti-infection therapy and restore the activity of LPS-induced osteoblasts. Moreover, miRNA-34a-5p was dramatically downregulated and SMAD2 was upregulated after LPS stimulation, while EHP resisted the inhibition of LPS by promoting miRNA-34a-5p, ALP, and BMP2 expressions. Whereas downregulation of miRNA-34a-5p reversed these effects. Silencing endogenous SMAD2 expression markedly promoted BMP2 and ALP activity and enhanced osteogenesis. Taken together, EHP restored LPS-induced bone loss by regulating miRNA-34a-5p levels and repressing its target gene SMAD2. EHP might be a potential adjuvant herbal remedy for the treatment of bone nonunion, and miRNA-34a-5p is a novel target for controlling bone and metabolic diseases.