Transfer of miR-877-3p via extracellular vesicles derived from dental pulp stem cells attenuates neuronal apoptosis and facilitates early neurological functional recovery after cerebral ischemia-reperfusion injury through the Bclaf1/P53 signaling pathway.

Cerebral ischemia-reperfusion injury (I/RI) is one of the principal pathogenic factors in the poor prognosis of ischemic stroke, for which current therapeutic options to enhance neurological recovery are notably insufficient. Dental pulp stem cell-derived extracellular vesicles (DPSC-EVs) have promising prospects in stroke treatment and the specific underlying mechanisms have yet to be fully elucidated. The present study observed that DPSC-EVs ameliorated the degree of cerebral edema and infarct volume by reducing the apoptosis of neurons. Furthermore, the miRNA sequencing and functional enrichment analysis identified that miR-877-3p as a key component in DPSC-EVs, contributing to neuroprotection and anti-apoptotic effects. Following target prediction and dual-luciferase assay indicated that miR-877-3p interacted with Bcl-2-associated transcription factor (Bclaf1) to play a function. The miR-877-3p inhibitor or Bclaf1 overexpression reversed the neuroprotective effects of DPSC-EVs. The findings reveal a novel therapeutic pathway where miR-877-3p, transferred via DPSC-EVs, confers neuroprotection against cerebral I/RI, highlighting its potential in promoting neuronal survival and recovery post-ischemia.

Dental pulp mesenchymal stem cell-derived exosomes inhibit neuroinflammation and microglial pyroptosis in subarachnoid hemorrhage via the miRNA-197-3p/FOXO3 axis.

BACKGROUND: Subarachnoid hemorrhage (SAH) is a severe stroke subtype that lacks effective treatment. Exosomes derived from human dental pulp stem cells (DPSCs) are a promising acellular therapeutic strategy for neurological diseases. However, the therapeutic effects of DPSC-derived exosomes (DPSC-Exos) on SAH remain unknown. In this study, we investigated the therapeutic effects and mechanisms of action of DPSC-Exos in SAH. MATERIALS AND METHODS: SAH was established using 120 male Sprague-Dawley rats. One hour after SAH induction, DPSC-Exos were administered via tail vein injection. To investigate the effect of DPSC-Exos, SAH grading, short-term and long-term neurobehavioral assessments, brain water content, western blot (WB), immunofluorescence staining, Nissl staining, and HE staining were performed. The role of miR-197-3p/FOXO3 in regulating pyroptosis was demonstrated through miRNA sequencing, bioinformatics analysis, and rescue experiments. The SAH model in vitro was established by stimulating BV2 cells with hemoglobin (Hb) and the underlying mechanism of DPSC-Exos was investigated through WB and Hoechst/PI staining. RESULTS: The expressions of pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α) were increased after SAH. DPSC-Exos alleviated brain edema and neuroinflammation by inhibiting the expression of FOXO3 and reducing NLRP3 inflammasome activation, leading to improved neurobehavioral functions at 24 h after SAH. In vitro, the expression of the NLRP3 inflammasome components (NLRP3 and caspase1-p20), GSDMD-N, and IL-18 was inhibited in BV2 cells pretreated with DPSC-Exos. Importantly, DPSC-Exos overexpressing miR-197-3p had a more obvious protective effect than those from NC-transfected DPSCs, while those from DPSCs transfected with the miR-197-3p inhibitor had a weaker protective effect. Functional studies indicated that miR-197-3p bound to the 3'-untranslated region of FOXO3, inhibiting its transcription. Furthermore, the overexpression of FOXO3 reversed the protective effects of miR-197-3p. CONCLUSIONS: DPSC-Exos inhibited activation of the NLRP3 inflammasome and related cytokine release via the miR-197-3p/FOXO3 pathway, alleviated neuroinflammation, and inhibited microglial pyroptosis. These findings suggest that using DPSC-Exos is a promising therapeutic strategy for SAH.

A novel polymer enabled by polymerized small molecule strategy for tumor photothermal and photodynamic therapy.

Photothermal therapy (PTT) and photodynamic therapy (PDT) are effective method for tumor treatment. However, the limited variety and quantity of photothermal agents (PTAs) and photosensitizer (PSs) are still major challenges. Moreover, the cell apoptosis mechanism induced by PDT and PTT is still elusive. A fused-ring small molecule acceptor-donor acceptor' donor-acceptor (A-DA'D-A) type of Y5 (Scheme 1) has a narrow band-gap and strong light absorption. Herein, we used Y5 to polymerize with thiophene unit to obtain polymer PYT based on polymerized small molecule strategy, and PYT nanoparticles (PYT NPs) was prepared via one-step nanoprecipitation strategy with DSPE-PEG2000. PYT NPs had excellent biocompatibility, good photostability, high photothermal conversion efficiency (67%) and reactive oxygen species (ROS) production capacity under 808 nm laser irradiation (PYT NPs + NIR). In vitro and in vivo experiments revealed that PYT NPs + NIR had the ability to completely ablate tumor cells. It was demonstrated that cell apoptosis induced by PYT NPs + NIR was closely related to mitochondrial damage. This study provides valuable guidance for constructing high-performance organic PTAs and PSs for tumor treatment. Scheme 1 PYT enabled by polymerized small molecule strategy for tumor photothermal and photodynamic therapy.

Profiling patient-reported symptom recovery from oesophagectomy for patients with oesophageal squamous cell carcinoma: a real-world longitudinal study.

PURPOSE: Oesophageal squamous cell carcinoma (ESCC) patients have severe symptom burden after oesophagectomy; however, longitudinal studies of symptom recovery after surgery are scarce. This study used longitudinal patient-reported outcome (PRO)-based symptoms to identify severe symptoms and profile symptom recovery from surgery in patients undergoing oesophagectomy. METHODS: Oesophageal cancer patients (N = 327) underwent oesophagectomy were consecutively included between April 2019 and March 2020. Data were extracted from the Sichuan Cancer Hospital's Esophageal Cancer Case Management Registration Database. Symptom assessment time points were pre-surgery and 1, 3, 5, 7, 14, 21, 30, and 90 days post-surgery using the Chinese version of the MD Anderson Symptom Inventory. And each symptom was rated on an 11-point scale, with 0 being 'not present' and 10 being 'as bad as you can imagine'. The symptom recovery trajectories were profiled using mixed effect models and Kaplan-Meier analysis. RESULTS: The most-severe symptoms on day 1 after oesophagectomy were pain, fatigue, dry mouth, disturbed sleep, and distress. The severity of symptoms peaked on day 1 after surgery. The top two symptoms were fatigue (mean: 5.44 [SD 1.88]) and pain (mean: 5.23 [SD 1.29]). Fatigue was more severe 90 days after surgery than at baseline (mean: 1.77 [SD 1.47] vs 0.65 [SD 1.05]; P < .0001). Disturbed sleep and distress persisted from pre-surgery to 90 days post-surgery; average sleep recovery time was up to 20 days, and 50.58% of patients had sleep disturbances 90 days post-surgery. CONCLUSIONS: Early post-operative pain management after oesophagectomy should be considered. Characteristics and intervention strategies of post-operative fatigue, distress, and disturbed sleep in oesophageal cancer patients warrant further studies.

[Emulsion liquid membrane extraction with D2EHPA mobile carrier Hirudinaria manillensis hirudin in experimental study].

OBJECTIVE: To study the extraction system of hirudin emulsion liquid membrane with the Poecilobdella manillensis as raw material, di-(2-ethylhexyl) phosphate (D2EHPA) as carrier, Span 80 as emulsifier, octane and D2EHPA mixed to constitute membrane solution, diluted HCl solutions as internal aqueous phase. METHOD: Using the orthogonal experiment to optimize the extraction conditions of hirudin reference substance such as membrane phase, internal aqueous phase volume ratio (MIPVR), external aqueous phase pH, internal aqueous phase pH, mobile carrier concentration and so on, and then using hirudin crude extracts to do purifying experiment, and gaining experimental samples. RESULT: The optimal conditions of hirudin extraction were as follows: MIPVR 10: 3, internal aqueous phase pH 2.6, external aqueous phase pH 3.4, the mass fraction of carrier D2EHPA 2%. In the optimal extraction conditions, when the initial concentration of hirudin was one anti-thrombin activity units (ATU) x mL(-1), ATU recovery rate of the reference substance was 83.06%. In the purifying experiment of crude extracts, ATU recovery rate was 82.99%, and the specific activity of sample was 3 289.48 the ATU x mg(-1). Discontinuous polyacrylamide gel electrophoresis and spectral scanning, the results showed that the purity and reference substance were considerable. CONCLUSION: The method of preparation hirudin was relatively simple, the purity of the experimental samples and ATU recovery were both high.

Electrochemistry-Induced Improvements of Mechanical Strength, Self-Healing, and Interfacial Adhesion of Hydrogels.

Hydrogels have demonstrated great potential in biomedical and engineering areas. To improve the physical performance, development of efficient physical/chemical protocols is essential. Herein, an electrochemistry functionalization strategy that is capable of enabling the functional improvements of hydrogel is reported. The electrochemistry functionalization is demonstrated on a hydrogel model of polyacrylamide (PAAm)@κ-carrageenan. The electrochemistry reaction generates metal ions (Fe3+ ) that migrate and coordinate with the sulfate groups of κ-carrageenan resulting in the prominent function improvements. In comparison with untreated PAAm@κ-carrageenan hydrogel, it can improve the mechanical strength by 7.37 times, and can increase the interfacial adhesion energy of the hydrogel on a glass surface from 0 to 1400 J m-2 , stronger than the bonding strength of tendons (adhesion energy: ≈800 J m-2 ). Two pieces of hydrogel strips integrate into an intact structure by the electrochemistry functionalization, where the healing efficiency reaches 100% in comparison to the untreated hydrogel. The most significant development is that it enables functional patterning on the hydrogel by the electrode assembly, which provides the hydrogel with modular sensitivity to external pressure. Therefore, it can be a general protocol for rapid generation of multifunctional hydrogels for biomedical and engineering developments.

[The effect of coaggregation of A.viscosus and P.gingivalis on the adherence of A.viscosus to hydroxyapatite].

PURPOSE: To examine the effect of coaggregation of A.viscosus and P.gingivalis on the adherence of A.viscosus to saliva-coated hydroxyapatite(S-HA). METHODS: The suspensions of A.viscosus and P.gingivalis were rotated and dropped on cover slips to visualize the coaggregation under scanning electron microscope(SEM). Different concentrations of P.gingivalis were mixed with A.viscosus radiolabeled with 10 uCi of [methyl-(3)H] thymedine per milliliter, then rotated for periods up to 1.5 hours at room temperature and the numbers of absorbed A.viscosus cells were determined by liquid scintillation. A.viscosus alone was used as the control, and its adhesion percent was set for 100%. One-way ANOVA with SPSS10.07 software package was used to analyze the effect of P.gingivalis on adherence of A.viscosus. RESULTS: Several cells of P.gingivalis could aggregate on the surface of A.viscosus. The adherence of A.viscosus to S-HA was promoted slightly when the concentration of P.gingivalis was rather small, then further increase in concentration of P.gingivalis resulted in a steady decline in the number of adherent A.viscosus, and the difference was significant (P<0.05). CONCLUSION: These results show that the adherence of A.viscosus to S-HA could be influenced by coaggregation of P.gingivalis and A.viscosus.

[Effect of A.viscosus on growth of P. gingivalis in vitro].

PURPOSE: To investigate the effect of A. viscosus on growth of P. gingivalis in vitro. METHODS: Groups as single-species(A.viscosus or P.gingivalis) and dual-species (A.viscosus and P.gingivalis) were incubated in BHI broth anaerobically at 37 degrees C, and growth curve of each group was monitored for 48 hours; Meanwhile the dual-species cultures of 2-hour interval were taken and the dilution was inoculated on BHI blood agar plates for 72 hours. The percentage of P. gingivalis was detected. BHI blood agar plates inoculated with P. gingivalis were prepared, and different concentrations of A. viscosus and its degerming supernatant were inoculated on the above plates in dropping method and incubated anaerobically for 72 hours. Zones of inhibition were measured and the data were analysed with SPSS 10.0 software package for one-way ANOVA. RESULTS: A. viscosus was in logarithmic phase of growth at 2-hour and reached plateau at 8-hour. P. gingivalis entered logarithmic phase at 12-hour and reached plateau at 32-hour. Co-culture group was in logarithmic phase of growth at 2-hour and reached plateau at 12-hour. The percentage of P.gingivalis decreased gradually on BHI plates (P<0.05) and at 8-hour no P. gingivalis colony was found. The average diameters (mm) of inhibition zones were 19.3, 17.4, 13.2, 9.8 when the concentration of A. viscosus applied was 1 x 10(9), 10(8), 10(7), 10(6) CFU/ml, respectively. There was significance difference among the four groups (P<0.05). The supernatant of A. viscosus didn't show restrain on plates with P. gingivalis. CONCLUSION: The study suggests that A. viscosus could inhibit the growth of P. gingivalis by nutrition competition in vitro.