N-acetyltransferase 10 regulates alphavirus replication via N4-acetylcytidine (ac4C) modification of the lymphocyte antigen six family member E (LY6E) mRNA.

Epitranscriptomic RNA modifications can regulate the stability of mRNA and affect cellular and viral RNA functions. The N4-acetylcytidine (ac4C) modification in the RNA viral genome was recently found to promote viral replication; however, the mechanism by which RNA acetylation in the host mRNA regulates viral replication remains unclear. To help elucidate this mechanism, the roles of N-acetyltransferase 10 (NAT10) and ac4C during the infection and replication processes of the alphavirus, Sindbis virus (SINV), were investigated. Cellular NAT10 was upregulated, and ac4C modifications were promoted after alphavirus infection, while the loss of NAT10 or inhibition of its N-acetyltransferase activity reduced alphavirus replication. The NAT10 enhanced alphavirus replication as it helped to maintain the stability of lymphocyte antigen six family member E mRNA, which is a multifunctional interferon-stimulated gene that promotes alphavirus replication. The ac4C modification was thus found to have a non-conventional role in the virus life cycle through regulating host mRNA stability instead of viral mRNA, and its inhibition could be a potential target in the development of new alphavirus antivirals.IMPORTANCEThe role of N4-acetylcytidine (ac4C) modification in host mRNA and virus replication is not yet fully understood. In this study, the role of ac4C in the regulation of Sindbis virus (SINV), a prototype alphavirus infection, was investigated. SINV infection results in increased levels of N-acetyltransferase 10 (NAT10) and increases the ac4C modification level of cellular RNA. The NAT10 was found to positively regulate SINV infection in an N-acetyltransferase activity-dependent manner. Mechanistically, the NAT10 modifies lymphocyte antigen six family member E (LY6E) mRNA-the ac4C modification site within the 3'-untranslated region (UTR) of LY6E mRNA, which is essential for its translation and stability. The findings of this study demonstrate that NAT10 regulated mRNA stability and translation efficiency not only through the 5'-UTR or coding sequence but also via the 3'-UTR region. The ac4C modification of host mRNA stability instead of viral mRNA impacting the viral life cycle was thus identified, indicating that the inhibition of ac4C could be a potential target when developing alphavirus antivirals.

Improved new bone formation capacity of hyaluronic acid-bone substitute compound in rat calvarial critical size defect.

BACKGROUND: Bone loss of residual alveolar ridges is a great challenge in the field of dental implantology. Deproteinized bovine bone mineral (DBBM) is commonly used for bone regeneration, however, it is loose and difficult to handle in clinical practice. Hyaluronic acid (HA) shows viscoelasticity, permeability and excellent biocompatibility. The aim of this study is to evaluate whether high-molecular-weight (MW) HA combined with DBBM could promote new bone formation in rat calvarial critical size defects (CSDs). MATERIALS AND METHODS: Rat calvarial CSDs (5 mm in diameter) were created. Rats (n = 45) were randomly divided into 3 groups: HA-DBBM compound grafting group, DBBM particles only grafting group and no graft group. Defect healing was assessed by hematoxylin-eosin staining and histomorphometry 2, 4 and 8 weeks postop, followed by Micro-CT scanning 8 weeks postop. Statistical analyses were performed by ANOVA followed by Tukey's post hoc test with P < 0.05 indicating statistical significance. RESULTS: All rats survived after surgery. Histomorphometric evaluation revealed that at 2, 4 and 8 weeks postop, the percentage of newly formed bone was significantly greater in HA-DBBM compound grafting group than in the other two groups. Consistently, Micro-CT assessment revealed significantly more trabecular bone (BV/TV and Tb.N) in HA-DBBM compound group than in the other two groups, respectively (P < 0.05). Moreover, the trabecular bone was significantly more continuous (Tb.Pf) in HA-DBBM compound group than in the other two groups, respectively (P < 0.05). CONCLUSION: HA not only significantly promoted new bone formation in rats calvarial CSDs but also improved the handling ability of DBBM.

Measurement of effective nonlinear coefficients in few-mode fibers.

Because of random mode coupling, the nonlinear coefficient in few-mode fibers (FMFs) is averaged to an effective value, which can be theoretically modeled and calculated by using the multi-mode Manakov equations. In this Letter, we experimentally measure the effective nonlinear coefficients in a 530-m-long FMF supporting two mode groups, namely, the $ {\rm LP}_{01} $LP01 and $ {\rm LP}_{11} $LP11 mode groups, by exploiting the self-phase and cross-phase modulations of pulsed fields. By using the nonlinear coefficient of the $ {\rm LP}_{01} $LP01 mode as a reference and comparing the spectral broadening of the pulsed fields, we obtain the intra-modal effective nonlinear coefficient of the $ {\rm LP}_{11} $LP11 mode and the inter-modal effective nonlinear coefficient between the $ {\rm LP}_{01} $LP01 and $ {\rm LP}_{11} $LP11 modes. The experimental results are in good agreement with the theoretical predictions of the multi-mode Manakov equations.

Simultaneous Temperature and Strain Measurements Using Polarization-Maintaining Few-Mode Bragg Gratings.

Simultaneous measurement of temperature and strain was demonstrated using a polarization-maintaining few-mode Bragg grating (PM-FMF-FBG) based on the wavelength and phase modulation of the even L P 11 mode. The wavelength shift sensitivity and the interrogated phase sensitivity of the temperature and strain were measured to be 10 pm·°C-1 and 0.73 pm·µÎµ-1 and -3.2 × 10-2 rad·°C-1 and 4 × 10-4 rad·µÎµ-1, respectively, with a discrimination efficiency of 98%. The polarization interference led to selective polarization excitation of the reflection spectra, and the calculated phase sensitivity agreed with the experimental results.

Investigation of the Pressure Gain Characteristics and Cycle Performance in Gas Turbines Based on Interstage Bleeding Rotating Detonation Combustion.

To further improve the cycle performance of gas turbines, a gas turbine cycle model based on interstage bleeding rotating detonation combustion was established using methane as fuel. Combined with a series of two-dimensional numerical simulations of a rotating detonation combustor (RDC) and calculations of cycle parameters, the pressure gain characteristics and cycle performance were investigated at different compressor pressure ratios in the study. The results showed that pressure gain characteristic of interstage bleeding RDC contributed to an obvious performance improvement in the rotating detonation gas turbine cycle compared with the conventional gas turbine cycle. The decrease of compressor pressure ratio had a positive influence on the performance improvement in the rotating detonation gas turbine cycle. With the decrease of compressor pressure ratio, the pressurization ratio of the RDC increased and finally made the power generation and cycle efficiency enhancement rates display uptrends. Under the calculated conditions, the pressurization ratios of RDC were all higher than 1.77, the decreases of turbine inlet total temperature were all more than 19 K, the power generation enhancements were all beyond 400 kW and the cycle efficiency enhancement rates were all greater than 6.72%.

21 spatial mode erbium-doped fiber amplifier for mode division multiplexing transmission.

We experimentally demonstrate a 12-mode group (21 spatial mode) cladding-pumped few-mode erbium-doped fiber amplifier. The differential modal gain (DMG) is dramatically reduced by using the double cladding erbium-doped fiber (DC-EDF) with a refractive index trench structure, which helps to tightly confine all 21 spatial modes in the core and to mitigate the bending loss. Our experimental results show that the DMG gain is about 3 dB when the average signal modal gain for all 12 mode groups is up to 15 dB across the C-band. Our method of using a DC-EDF with a properly designed trench structure can be used to develop gain equalized few-mode amplifiers supporting more spatial modes.

Influences of Schneiderian membrane conditions on the early outcomes of osteotome sinus floor elevation technique: a prospective cohort study in the healing period.

OBJECTIVES: To radiographically investigate early outcomes of osteotome sinus floor elevation in the healing phase utilizing cone beam computed tomography and evaluate influences of Schneiderian membrane conditions. MATERIAL AND METHODS: One hundred patients were consecutively recruited for osteotome sinus floor elevation (OSFE) surgery using deproteinized bone mineral. CBCT was taken prior to (T0), immediately post-operatively (T1), and after the healing period (T2). Linear and volumetric measurements of the elevated region from T0 to T1 were performed for evaluation on computed tomography (CT). RESULTS: Osteotome sinus floor elevation were performed in 100 patients. One implant of each patient was selected. Mean residual bone height (RH) was 7.21 ± 1.12 mm. Mean sinus floor elevation height (SE) was 4.81 ± 0.75 mm. The mean endo-sinus bone gain after the healing period was 3.25 ± 0.83 mm. Pre-opterative CBCT scans revealed that 72 patients had a normal sinus membrane in osteotome region, 13 patients presented with flat thickened mucosa and 15 patients with antral pseudocysts. There is no significant difference in sinus mucosa elevation height, bone graft volume and new bone formation in group of Thickening membrane and Antral pseudocysts compared with normal. CONCLUSIONS: The radiographical results show that OSFE is a safe and predictable surgical procedure in residual bone height of 7.21 ± 1.12 mm. Mild flat thickening (>2 and <5 mm) and antral pseudocysts in a small size without clinical symptoms may not be contraindications to OSFE surgery.

Transformation of erythromycin during secondary effluent soil aquifer recharging: Removal contribution and degradation path.

Erythromycin (ERY), a widely used antibiotic, has recently been detected in municipal secondary effluents and poses serious threats to human health during wastewater reusing. In this study, the removal, fate, and degradation pathway of ERY in secondary effluent during soil aquifer treatment was evaluated via laboratory-scale SAT tests. Up to a 92.9% reduction of ERY in synthetic secondary effluent was observed in 1.0m depth column system, which decreased to 64.7% when recharged with wastewater treatment plant secondary effluent. XRD-fractionation results demonstrated that the transphilic acid and hydrophobic acid fractions in secondary effluent compete for the adsorption sites of the packed soil and lead to a declined ERY removal. Moreover, aerobic biodegradation was the predominant role for ERY removal, contributing more than 60% reduction of ERY when recharged with synthetic secondary effluent. Destruction of 14-member macrocyclic lactone ring and breakdown of two cyclic sugars (l-cladinose and d-desosamine) were main removal pathways for ERY degradation, and produced six new intermediates.

Characteristics and Tissue Regeneration Properties of Gingiva-Derived Mesenchymal Stem Cells.

Mesenchymal stem cells (MSCs) have 2 specific characteristics: self-renewal and multi- or pluripotency. Extensive studies have demonstrated the regenerative capability of MSCs both in vitro and in vivo. Gingiva-derived MSCs (GMSCs) represent a unique population of MSCs that can be easily isolated and obtained. GMSCs, which maintain a normal karyotype and telomerase activity in long-term cultures, display a stable phenotype and rapidly proliferate in vitro. In addition, GMSCs can be induced to differentiate into osteogenic, chondrogenic, and adipogenic lineages. Therefore, GMSCs are a promising alternative cell source for tissue regeneration in dentistry. In this article we review studies of the characterization, differentiation capacities, and regenerative role of GMSCs derived from the gingiva of humans and other species, focusing on the mechanisms of differentiation and tissue regeneration of human GMSCs. We anticipate that GMSC-based therapies will significantly contribute to regenerative medicine for the treatment of human dental diseases and improve human health.

Effect of molecular weight and concentration of hyaluronan on cell proliferation and osteogenic differentiation in vitro.

Hyaluronan (HA), the simplest glycosaminoglycan and a major component of the extracellular matrix, exists in various tissues. It is involved in some critical biological procedures, including cellular signaling, cell adhesion and proliferation, and cell differentiation. The effect of molecular weight (MW) and concentration of HA on cell proliferation and differentiation was controversial. In this study, we investigated the effect of MW and concentration of HA on the proliferation and osteogenic differentiation of rabbit bone marrow-derived stem cells in vitro. Results showed that high MW HA decreased the cell adhesion rate in a concentration-dependant manner. The cell adhesion rate was decreased by increasing MW of HA. Cell proliferation was significantly enhanced by low MW HA (P < 0.05). The factorial analysis indicated that MW and concentration had an interactive effect on the cell adhesion rate and cell proliferation (P < 0.05). High MW HA increased the mRNA expressions of ALP, RUNX-2 and OCN. The higher the MW was, the higher the mRNA expressions were. The factorial analysis indicated that MW and concentration had an interactive effect on ALP mRNA expression (P < 0.05). HA of higher MW and higher concentration promoted bone formation. These findings provide some useful information in understanding the mechanism underlying the effect of MW and concentration of HA on cell proliferation and differentiation.

Time-division-multiplexed few-mode passive optical network.

We demonstrate the first few-mode-fiber based passive optical network, effectively utilizing mode multiplexing to eliminate combining loss for upstream traffic. Error-free performance has been achieved for 20-km low-crosstalk 3-mode transmission in a commercial GPON system carrying live Ethernet traffic. The alternative approach of low modal group delay is also analyzed with simulation results over 10 modes.

Generation of frequency degenerate twin photons in pulse pumped fiber optical parametric amplifiers: influence of background noise.

Using a Sagnac fiber loop functions as a deterministic splitter of photon pairs produced by the frequency degenerate four wave mixing, we show that the background noise of the degenerate photon pairs is contributed by both Raman scattering and frequency non-degenerate four wave mixing. To improve the purity of photon pairs in the high gain regime, in addition to suppressing the noise photons by cooling the nonlinear fiber and by optimizing the detuning between the frequencies of the pump and photon pairs, the walk-off effect of the two pulsed pump fields should be mitigated by managing the dispersion of the fiber. Our investigation is not only the first step towards the generation of multi-mode squeezed vacuum in fiber optical parametric amplifiers pumped with pulsed lights, but also contributes to improving the purity of the fiber sources of degenerate photon pairs.

A novel approach in biomedical engineering: The use of polyvinyl alcohol hydrogel encapsulating human umbilical cord mesenchymal stem cell-derived exosomes for enhanced osteogenic differentiation and angiogenesis in bone regeneration.

Developing effective methods for alveolar bone defect regeneration is a significant challenge in orthopedics. Exosomes from human umbilical cord mesenchymal stem cells (HUMSC-Exos) have shown potential in bone repair but face limitations due to undefined application methods and mechanisms. To address this, HUMSC-Exos were encapsulated in polyvinyl alcohol (PVA) hydrogel (Exo@PVA) to create a novel material for alveolar bone repair. This combination enhanced the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) more effectively than Exos alone. Additionally, Exo@PVA significantly improved alveolar bone regeneration and defect repair in rats. The microRNA-21-5p (miR-21-5p) in Exo@PVA, identified through the GEO database and analyzed via in silico methods, played a crucial role. miR-21-5p promoted BMSC osteogenic differentiation by inhibiting WWP1-mediated KLF5 ubiquitination and enhanced HUVEC angiogenesis by targeting ATP2B4. These findings underscore the potential of an Exo-based approach with PVA hydrogel scaffolds for bone defect repair, operating through the miR-21-5p/WWP1/ATP2B4 signaling axis.

Accuracy analysis of robotic-assisted immediate implant placement: A retrospective case series.

OBJECTIVES: This study aimed to investigate the accuracy of a robotic computer-assisted implant surgery (r-CAIS) for immediate implant placement. METHODS: Patients requiring immediate implant placement in the maxillary anterior region were enrolled for r-CAIS. Before surgery, the patients underwent a cone beam computed tomography (CBCT) scan with a positioning marker. Virtual implant placement position and drilling sequences were planned. Following spatial registration and calibration, the implants were placed with the robotic system under supervision. A postoperative CBCT was taken to control the actual implant positions. The DICOM data of the virtually planned and the actually placed implant were superimposed and registered through the accuracy verification software of the robotic system. The accuracy was calculated automatically. The deviation at the mesial-distal, labial-palatal, and apico-coronal directions were recorded. RESULTS: Fifteen patients with 20 implants were included. No adverse surgical events or postoperative complications were reported. The global platform, apex, and angular deviation were 0.75 ± 0.20 mm (95 % CI: 0.65 to 0.84 mm), 0.70 ± 0.27 mm (95 % CI: 0.57 to 0.82 mm), and 1.17 ± 0.73° (95 % CI: 0.83 to 1.51°), respectively. Moreover, the vertical platform and apex deviation were 0.50 ± 0.31 mm, (95 % CI: 0.35 to 0.64 mm) and 0.48 ± 0.32 mm, (95 % CI: 0.33 to 0.63 mm), respectively. All the placed implant positions were further labial and apical than the planned ones, respectively. CONCLUSIONS: High accuracy of immediate implant placement was achieved with the robotic system. CLINICAL SIGNIFICANCE: Our study provided evidence to support the potential of the robotic system in implant placement, even in challenging scenarios.

Prognostic value of ultrasound in early arterial complications post liver transplant.

Liver transplantation is the primary therapeutic intervention for end-stage liver disease. However, vascular complications, particularly those involving the hepatic artery, pose significant risks to patients. The clinical manifestations associated with early arterial complications following liver transplantation are often nonspecific. Without timely intervention, these complications can result in graft failure or patient mortality. Therefore, early diagnosis and the formulation of an optimal treatment plan are imperative. Ultrasound examination remains the predominant imaging modality for detecting complications post liver transplantation. This article comprehensively reviews common causes and clinical presentations of early hepatic artery complications in the post-transplantation period and delineates abnormal sonographic findings for accurate diagnosis of these conditions. Overall, ultrasound offers the advantages of convenience, safety, effectiveness, and non-invasiveness. It enables real-time, dynamic, and precise evaluation, making it the preferred diagnostic method for post-liver transplantation assessments.

Prediction of hepatic artery occlusion after liver transplantation by ultrasound characteristics and clinical risk factors.

BACKGROUND: Hepatic artery occlusion (HAO) after liver transplantation (LT) is a devastating complication, resulting in early graft loss and reduced overall survival. Ultrasound is an established assessment method for HAO in patients following LT, especially those with complex hepatic artery reconstruction. AIM: To investigate the ultrasound characteristics and analyze the risk factors associated with HAO in patients after LT. METHODS: We retrospectively analyzed the ultrasound characteristics and the clinic risk factors associated with HAO in 400 adult LT patients who were enrolled and treated at the Third People's Hospital of Shenzhen between November 2016 and July 2022. Fourteen patients diagnosed with acute HAO (A-HAO) by surgery and fifteen diagnosed with chronic HAO (C-HAO) were included. A control group of 33 patients without HAO complications during the same period were randomly selected using a random number table. All patients underwent an ultrasonography examination. Parameters including resistance index (RI), peak systolic velocity (PSV), and portal vein velocity (PVV) were compared across the groups. Additionally, basic clinical data were collected for all patients, including gender, age, primary diagnosis, D-dimer concentration, total operation time, cold ischemia time, hot ischemia time, intraoperative blood loss and transfusion, intraoperative urine volume, infusion, model for end-stage liver disease (MELD) score, and whether complex hepatic artery reconstructions were performed. Furthermore, risk factors influencing HAO formation after LT were analyzed. RESULTS: Compared to the non-HAO group, PVV and RI were higher in the A-HAO group, while PSV was lower. Conversely, both PSV and RI were lower in the C-HAO group compared to the non-HAO group. The proportion of patients undergoing complex hepatic artery reconstructions and the gamma-glutamyltransferase (GGT) level before occlusion were significantly higher in the A-HAO group compared to the non-HAO group. However, there were no distinct differences between the two groups in D-dimer, MELD score, pre-occlusion alanine transaminase and aspartate transaminase levels, or intraoperative conditions. CONCLUSION: Ultrasound features of the hepatic artery before occlusion are significantly associated with postoperative HAO development. Additionally, complex hepatic artery reconstructions, defined as revascularization of the graft requiring additional anastomosis between donor hepatic arteries, constitute a risk factor for A-HAO. Besides, abnormal pre-occlusion GGT elevation is an important biochemical indicator. Therefore, ultrasound examination serves as an important tool for screening HAO, especially in patients with the identified risk factors.

Accuracy of dynamic navigation compared to static surgical guides and the freehand approach in implant placement: a prospective clinical study.

BACKGROUND: Computer-guided implant surgery has improved the quality of implant treatment by facilitating the placement of implants in a more accurate manner. This study aimed to assess the accuracy of implant placement in a clinical setting using three techniques: dynamic navigation, static surgical guides, and freehand placement. We also investigated potential factors influencing accuracy to provide a comprehensive evaluation of each technique's advantages and disadvantages. MATERIALS AND METHODS: Ninety-four implants in 65 patients were included in this prospective study. Patients were randomly assigned to one of three groups: dynamic navigation, static surgical guides, or freehand placement. Implants were placed using a prosthetically oriented digital implant planning approach, and postoperative CBCT scans were superimposed on preoperative plans to measure accuracy. Seven deviation values were calculated, including angular, platform, and apical deviations. Demographic and consistency analyses were performed, along with one-way ANOVA and post-hoc tests for deviation values. RESULTS: The mean global platform, global apical, and angular deviations were 0.99 mm (SD 0.52), 1.14 mm (SD 0.56), and 3.66° (SD 1.64°) for the dynamic navigation group; 0.92 mm (SD 0.36), 1.06 mm (SD 0.47), and 2.52° (SD 1.18°) for the surgical guide group; and 1.36 mm (SD 0.62), 1.73 mm (SD 0.66), and 5.82° (SD 2.79°) for the freehand group. Both the dynamic navigation and surgical guide groups exhibited statistically significant differences in all values except depth deviations compared to the freehand group (p < 0.05), whereas only the angular deviation showed a significant difference between the dynamic navigation and surgical guide groups (p = 0.002). CONCLUSION: Our findings highlight the superior accuracy and consistency of dynamic navigation and static surgical guides compared to freehand placement in implant surgery. Dynamic navigation offers precision and flexibility. However, it comes with cost and convenience considerations. Future research should focus on improving its practicality. TRIAL REGISTRATION: This study was retrospectively registered at the Thai Clinical Trials Register-Medical Research Foundation of Thailand (MRF) with the TCTR identification number TCTR20230804001 on 04/08/2023. It was also conducted in accordance with the Declaration of Helsinki and approved by the institutional ethics committee at the Xian Jiaotong University Hospital of Stomatology, Xian, China (xjkqII[2021] No: 043). Written informed consent was obtained from all participants.

Tanreqing injection inhibits influenza virus replication by promoting the fusion of autophagosomes with lysosomes: An integrated pharmacological study.

ETHNOPHARMACOLOGICAL RELEVANCE: Tanreqing injection (TRQ) is widely used, traditional Chinese medicine (TCM) injection used in China to treat respiratory infections. Modern pharmacological studies have confirmed that TRQ can protect against influenza viruses. However, the mechanism by which TRQ inhibits influenza viruses remains unclear. AIM OF THE STUDY: To explore the therapeutic effects and possible mechanisms of TRQ inhibition by the influenza virus. MATERIALS AND METHODS: Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) was used to determine the chemical composition of TRQ. Isobaric tags for relative and absolute quantification (iTRAQ) were used to define differential proteins related to TRQ inhibition of viruses. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed for functional annotation. For experimental validation, we established an in vitro model of the influenza virus infection by infecting A549 cells with the virus. The detection of the signaling pathway was carried out through qPCR, western blotting,and immunofluorescence. RESULTS: Fifty one components were identified using UPLC/Q-TOF MS. We confirmed the inhibitory effect of TRQ on influenza virus replication in vitro. Ninety nine differentially expressed proteins related to the inhibitory effect of TRQ were identified using iTRAQ. KEGG functional enrichment analysis showed that the TRQ may inhibit influenza virus replication by affecting autophagy. Through network analysis, 29 targets were selected as major targets, and three key targets, HSPA5, PARP1, and GAPDH, may be the TRQ targets affecting autophagy. In vitro experiments showed that TRQ inhibits influenza virus replication by interfering with the expression and localization of STX17 and VAMP8 proteins, thereby promoting the fusion of autophagosomes with lysosomes. CONCLUSION: TRQ inhibits influenza virus replication by promoting the fusion of autophagosomes with lysosomes. We additionally established potential gene and protein targets which are affected by TRQ. Therefore, our findings provide new therapeutic targets and a foundation further studies on influenza treatment with TRQ.

Flexible nano-liposomes-encapsulated recombinant UL8-siRNA (r/si-UL8) based on bioengineering strategy inhibits herpes simplex virus-1 infection.

Herpes simplex virus-1 (HSV-1) infection can cause various diseases and the current therapeutics have limited efficacy. Small interfering RNA (siRNA) therapeutics are a promising approach against infectious diseases by targeting the viral mRNAs directly. Recently, we employed a novel tRNA scaffold to produce recombinant siRNA agents with few natural posttranscriptional modifications. In this study, we aimed to develop a specific prodrug against HSV-1 infection based on siRNA therapeutics by bioengineering technology. We screened and found that UL8 of the HSV-1 genome was an ideal antiviral target based on RNAi. Next, we used a novel bio-engineering approach to manufacture recombinant UL8-siRNA (r/si-UL8) in Escherichia coli with high purity and activity. The r/si-UL8 was selectively processed to mature si-UL8 and significantly reduced the number of infectious virions in human cells. r/si-UL8 delivered by flexible nano-liposomes significantly decreased the viral load in the skin and improved the survival rate in the preventive mouse zosteriform model. Furthermore, r/si-UL8 also effectively inhibited HSV-1 infection in a 3D human epidermal skin model. Taken together, our results highlight that the novel siRNA bioengineering technology is a unique addition to the conventional approach for siRNA therapeutics and r/si-UL8 may be a promising prodrug for curing HSV-1 infection.

PF127 Hydrogel-Based Delivery of Exosomal CTNNB1 from Mesenchymal Stem Cells Induces Osteogenic Differentiation during the Repair of Alveolar Bone Defects.

Pluronic F127 (PF127) hydrogel has been highlighted as a promising biomaterial for bone regeneration, but the specific molecular mechanism remains largely unknown. Herein, we addressed this issue in a temperature-responsive PF127 hydrogel loaded with bone marrow mesenchymal stem cells (BMSCs)-derived exosomes (Exos) (PF127 hydrogel@BMSC-Exos) during alveolar bone regeneration. Genes enriched in BMSC-Exos and upregulated during the osteogenic differentiation of BMSCs and their downstream regulators were predicted by bioinformatics analyses. CTNNB1 was predicted to be the key gene of BMSC-Exos in the osteogenic differentiation of BMSCs, during which miR-146a-5p, IRAK1, and TRAF6 might be the downstream factors. Osteogenic differentiation was induced in BMSCs, in which ectopic expression of CTNNB1 was introduced and from which Exos were isolated. The CTNNB1-enriched PF127 hydrogel@BMSC-Exos were constructed and implanted into in vivo rat models of alveolar bone defects. In vitro experiment data showed that PF127 hydrogel@BMSC-Exos efficiently delivered CTNNB1 to BMSCs, which subsequently promoted the osteogenic differentiation of BMSCs, as evidenced by enhanced ALP staining intensity and activity, extracellular matrix mineralization (p < 0.05), and upregulated RUNX2 and OCN expression (p < 0.05). Functional experiments were conducted to examine the relationships among CTNNB1, microRNA (miR)-146a-5p, and IRAK1 and TRAF6. Mechanistically, CTNNB1 activated miR-146a-5p transcription to downregulate IRAK1 and TRAF6 (p < 0.05), which induced the osteogenic differentiation of BMSCs and facilitated alveolar bone regeneration in rats (increased new bone formation and elevated BV/TV ratio and BMD, all with p < 0.05). Collectively, CTNNB1-containing PF127 hydrogel@BMSC-Exos promote the osteogenic differentiation of BMSCs by regulating the miR-146a-5p/IRAK1/TRAF6 axis, thus inducing the repair of alveolar bone defects in rats.