Methane production and lignocellulosic degradation of wastes from rice, corn and sugarcane by natural anaerobic fungi-methanogens co-culture.

Biomass from agriculture, forestry, and urban wastes is a potential renewable organic resource for energy generation. Many investigations have demonstrated that anaerobic fungi and methanogens could be co-cultured to degrade lignocellulose for methane generation. Thus, this study aimed to evaluate the effect of natural anaerobic fungi-methanogens co-culture on the methane production and lignocellulosic degradation of wastes from rice, corn and sugarcane. Hu sheep rumen digesta was used to develop a natural anaerobic fungi-methanogen co-culture. The substrates were rice straw (RS), rich husk (RH), corn stover (CS), corn cobs (CC), and sugarcane baggage (SB). Production of total gas and methane, metabolization rate of reducing sugar, glucose, and xylose, digestibility of hemicellulose and cellulose, activity of carboxymethylcellulase and xylanase, and concentrations of total acid and acetate were highest (P < 0.05) in CC, moderate (P < 0.05) in RS and CS, and lowest (P < 0.05) in SB and RH. The pH, lactate and ethanol were lowest (P < 0.05) in CC, moderate (P < 0.05) in RS and CS, and lowest (P < 0.05) SB and RH. Formate was lowest (P < 0.05) in CC, RS and CS, moderate (P < 0.05) in SB, and lowest (P < 0.05) in RH. Therefore, this study indicated that the potential of methane production and lignocellulosic degradation by natural anaerobic fungi-methanogens co-culture were highest in CC, moderate in RS and CS, and lowest in SB and RH.

BML-111 inhibit H2O2-induced pyroptosis and osteogenic dysfunction of human periodontal ligament fibroblasts by activating the Nrf2/HO-1 pathway.

BACKGROUND: Periodontitis is a common and harmful chronic inflammatory oral disease, characterized by the destruction of periodontal soft and hard tissues. The NLRP3 inflammasome-related pyroptosis and human periodontal ligament fibroblasts (hPDLFs) osteogenic dysfunction are involved in its pathogenesis. Studies have shown that lipoxin A4 is an endogenous anti-inflammatory mediator and BML-111 is a lipoxin A4 analog, which was found to have potent and durable anti-inflammatory effects in inflammatory diseases, but the mechanism remains unclear. The purpose of this study was to investigate whether BML-111 inhibits H2O2-induced dysfunction of hPDLFs, attenuates inflammatory responses, and identifies the underlying mechanisms. METHODS: The oxidative stress model was established with H2O2, and the cell proliferation activity was measured by CCK-8. ALP staining and alizarin red staining were used to detect the osteogenic differentiation capacity of cells; flow cytometry and ELISA were used to detect cell pyroptosis; we explored the effect of BML-111 on hPDLFs under oxidative stress by analyzing the results of PCR and Western blotting. The Nrf2 inhibitor ML385 was added to further identify the target of BML-111 and clarify its mechanism. RESULTS: BML-111 can alleviate the impaired cell proliferation viability induced by H2O2. H2O2 treatment can induce NLRP3 inflammasome-related pyroptosis, impairing the osteogenic differentiation capacity of hPDLFs. BML-111 can effectively alleviate H2O2-induced cellular dysfunction by activating the Nrf2/HO-1 signaling pathway. CONCLUSION: The results of this study confirmed the beneficial effects of BML-111 on H2O2-induced NLRP3 inflammasome-related pyroptosis in hPDLFs, and BML-111 could effectively attenuate the impaired osteogenic differentiation function. This beneficial effect is achieved by activating the Nrf2/HO-1 signaling pathway, therefore, our results suggest that BML-111 is a potential drug for the treatment of periodontitis.

Virtual Surgical Planning and Three-Dimensional Printing to Aid the Anatomical Reduction of an Old Malunited Fracture of the Mandible.

ABSTRACT: Mandible fracture is a common injury in maxillofacial surgery. It causes not only maxillofacial dysfunction but also facial deformities. Malunited fractures of the mandible have been a vast challenge in clinical treatment due to the misalignment of the broken ends and the occurrence of occlusal disorders. This case report describes using virtual surgical planning and three-dimensional printing to treat a patient with malunited fracture of the mandible. Failing to perform mandibular surgery due to severe brain trauma after the car accident, the patient got malunited healing of mandible. The authors applied virtual surgical planning to perform preoperative analysis and surgical design on this patient, three-dimensional printing to fabricate occlusal plate, and models of the preoperative and postoperative mandible to guide the operation. Finally, the authors achieved the reduction and reconstruction of the mandible with satisfactory clinical results.

Acute Kidney Injury Secondary to Severe Hand, Foot and Mouth Disease Caused by Enterovirus-A71: Hypertension Is a Common.

Hand, foot and mouth disease (HFMD) is a common infectious disease among children, caused primarily by human enterovirus-A71 (EV-A71) and coxsackievirus-A16 (CV-A16). To date, only two case reports mention that renal involvement can be secondary to or coexisting with CV-A16-associated HFMD. In the present report, we describe a 10-year-old girl who was infected with EV-A71 and subsequently developed a definite acute kidney injury (AKI), mainly based on the characteristic rash, virus isolation, eyelid edema, hypertension, decreased urine output, mild proteinuria and impaired renal function. She was treated with intravenous ribavirin, immunoglobulin, oral administration of nifedipine and ramipril. After 7 days of intensive observations, she recovered fully. Hypertension is a common feature in both HFMD and AKI. On one hand, hypertension serves as a risk factor for severe HFMD; on the other hand, hypertension induces AKI onset and is also deteriorated by AKI.

Microplastic accumulation in groundwater: Data-scaled insights and future research.

Given that microplastics (MPs) in groundwater have been concerned for risks to humans and ecosystems with increased publications, a Contrasting Analysis of Scales (CAS) approach is developed by this study to synthesize all existing data into a hierarchical understanding of MP accumulation in groundwater. Within the full data of 386 compiled samples, the median abundance of MPs in Open Groundwater (OG) and Closed Groundwater (CG) were 4.4 and 2.5 items/L respectively, with OG exhibiting a greater diversity of MP colors and larger particle sizes. The different pathways of MP entry (i.e., surface runoff and rock interstices) into OG and CG led to this difference. At the regional scale, median MP abundance in nature reserves and landfills were 17.5 and 13.4 items/L, respectively, all the sampling points showed high pollution load risk. MPs in agricultural areas exhibited a high coefficient of variation (716.7%), and a median abundance of 1.0 items/L. Anthropogenic activities at the regional scale are the drivers behind the differentiation in the morphological characteristics of MPs, where groundwater in residential areas with highly toxic polymers (e.g., polyvinylchloride) deserves prolonged attention. At the local scale, the transport of MPs is controlled by groundwater flow paths, with a higher abundance of MP particles downstream than upstream, and MPs with regular surfaces and lower resistance (e.g., pellets) are more likely to be transported over long distances. From the data-scaled insight this study provides on the accumulation of MPs, future research should be directed towards network-based observation for groundwater-rich regions covered with landfills, residences, and agricultural land.

CA1 Modulates the Osteogenic Differentiation of Dental Follicle Stem Cells by Activating the BMP Signaling Pathway In Vitro.

BACKGROUND: Carbonic anhydrase 1 (CA1) has been found to be involved in osteogenesis and osteoclast in various human diseases, but the molecular mechanisms are not completely understood. In this study, we aim to use siRNA and lentivirus to reduce or increase the expression of CA1 in Dental follicle stem cells (DFSCs), in order to further elucidate the role and mechanism of CA1 in osteogenesis, and provide better osteogenic growth factors and stem cell selection for the application of bone tissue engineering in alveolar bone fracture transplantation. METHODS: The study used RNA interference and lentiviral vectors to manipulate the expression of the CA1 gene in DFSCs during in vitro osteogenic induction. The expression of osteogenic marker genes was evaluated and changes in CA1, alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), and Bone morphogenetic proteins (BMP2) were measured using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB). The osteogenic effect was assessed through Alizarin Red staining. RESULTS: The mRNA and protein expression levels of CA1, ALP, RUNX2, and BMP2 decreased distinctly in the si-CA1 group than other groups (p < 0.05). In the Lentivirus-CA1 (LV-CA1) group, the mRNA and protein expressions of CA1, ALP, RUNX2, and BMP2 were amplified to varying degrees than other groups (p < 0.05). Apart from CA1, BMP2 (43.01%) and ALP (36.69%) showed significant upregulation (p < 0.05). Alizarin red staining indicated that the LV-CA1 group produced more calcified nodules than other groups, with a higher optical density (p < 0.05), and the osteogenic effect was superior. CONCLUSIONS: CA1 can impact osteogenic differentiation via BMP related signaling pathways, positioning itself upstream in osteogenic signaling pathways, and closely linked to osteoblast calcification and ossification processes.

Case report: Plastic bronchitis associated with Bordetella parapertussis.

RATIONALE: Bordetella parapertussis caused by a severe infection is rare in clinical practice. Here, we report a case of plastic bronchitis (PB). PATIENT CONCERNS: A 4-year-old girl with a 2-day history of fever, paroxysmal cough, and subconjunctival hemorrhage. DIAGNOSES: The diagnoses were (1) B parapertussis , (2) pulmonary atelectasis, and (3) PB. INTERVENTIONS: The patient received azithromycin and underwent bronchoscopy. OUTCOMES: Symptoms disappeared after treatment. The patient had an outpatient follow-up of 2 months without respiratory symptoms. LESSONS: PB can lead to respiratory failure if not intervened in the early stages.

Remodeling of the periodontal ligament and alveolar bone during axial tooth movement in mice with type 1 diabetes.

Objectives: To observe the elongation of the axial tooth movement in the unopposed rodent molar model with type 1 diabetes mellitus and explore the pathological changes of periodontal ligament and alveolar bone, and their correlation with tooth axial movement. Methods: The 80 C57BL/6J mice were randomly divided into the streptozotocin(STZ)-injected group (n = 50) and the control group (n = 30). Mice in the streptozotocin(STZ)-injected group were injected intraperitoneal with streptozotocin (STZ), and mice in the control group were given intraperitoneal injection of equal doses of sodium citrate buffer. Thirty mice were randomly selected from the successful models as the T1DM group. The right maxillary molar teeth of mice were extracted under anesthesia, and allowed mandibular molars to super-erupt. Mice were sacrificed at 0, 3, 6,9, and 12 days. Tooth elongation and bone mineral density (BMD) were evaluated by micro-CT analysis(0,and 12 days mice). Conventional HE staining, Masson staining and TRAP staining were used to observe the changes in periodontal tissue(0, 3, 6, 9, and 12 days mice). The expression differences of SPARC, FGF9, BMP4, NOGGIN, and type I collagen were analyzed by RT-qPCR. Results: After 12 days of tooth extraction, our data showed significant super-eruption of mandibular mouse molars of the two groups. The amount of molar super-eruption in the T1DM group was 0.055mm( ± 0.014mm), and in the control group was 0.157( ± 0.017mm). The elongation of the T1DM mice was less than that of the control mice(P<0.001). It was observed that the osteoclasts and BMD increased gradually in both groups over time. Compared with the control group, the collagen arrangement was more disordered, the number of osteoclasts was higher (P<0.05), and the increase of bone mineral density was lower(2.180 ± 0.007g/cm3 vs. 2.204 ± 0.006g/cm3, P<0.001) in the T1DM group. The relative expression of SPARC, FGF9, BMP4, and type I collagen in the two groups increased with the extension of tooth extraction time while NOGGIN decreased. The relative expression of all of SPARC, FGF9, BMP4, and type I collagen in the T1DM group were significantly lower, and the expression of NOGGIN was higher than that in the control group (P<0.05). Conclusion: The axial tooth movement was inhibited in type 1 diabetic mice. The result may be associated with the changes of periodontal ligament osteoclastogenic effects and alveolar bone remodeling regulated by the extracellular matrix and osteogenesis-related factors.

Comparative study of stress characteristics around the adjacent teeth tissues during insertion of mini-screws with different insertion angles: A three-dimensional finite element study.

With a limited alveolar bone position, there is a high risk that mini-screws (MS) implants could cause damage to the adjacent teeth. To reduce this damage, the position and tilt angle of the MS must be optimized. The aim of this study was to assess the effect of MS implantation angle on the stress exerted on adjacent periodontal membrane and roots. A three-dimensional finite element model containing dentition, periodontal ligament, jaw and MS were established based on the CBCT images and MS scanning data. The MS was first inserted perpendicular to the surface of the bone at specific locations and then tilted at an angle of 10° and 20° to the mesial and distal teeth, respectively. The stress distribution in the periodontal tissue of the adjacent teeth was analyzed after MS implantation at different angles.The stress on the adjacent tooth root and periodontal ligament was most uniformly distributed when the MS was inserted vertically. It changed 9.4-97.7% when the axis of MS was tilted at 10-degree and 20-degree angles from the point of vertical insertion. The stresses experienced by the periodontal ligament and the root are similar. When the horizontal angle of the MS insertion was changed, the MS was closer to the adjacent tooth, resulting in greater stress near the PDL and root. It was recommended to insert the MS vertically into the alveolar bone surface to avoid root damage due to excessive stress.

Introduction of Accurate Measurement of the Alveolar Bone Height in Bone Grafting Area after Sinus Lift.

BACKGROUND: Sinus lift via the alveolar crest approach has been proven to solve inadequate alveolar bone height perfectly in the posterior region of the maxilla. Nevertheless, the surgical area cannot be directly observed, imaging methods are necessary to reveal whether the surgery has achieved the expected results. Cone-Beam Computed Tomography (CBCT) images are usually taken before surgery and at the completion of final pre-repair osseointegration and evaluate the effect of maxillary sinus lift by measuring the alveolar bone height in the bone graft area. However, there is no uniform and recognized method to accurately measure the alveolar bone height in the bone grafting area before and after maxillary sinus lift. Therefore, the authors propose a simple, accurate, and reproducible method for the measurement of alveolar bone height in maxillary sinus lift on CBCT, which will be an innovation. METHODS: 30 implant cases (30 patients) were included in this study. The authors create a uniformed measurement surface in the CBCT axial interface to ensure consistency at the completion of final prerepair osseointegration and before surgery. Then changes in alveolar bone height in the bone grafting area before and after sinus lift were measured on CBCT coronal and arch planes. Finally, for the purpose of verification of repeatability, three dentists from our department were randomly selected to measure and record the above CBCT data at three different time points. RESULTS: The data showed that in the CBCT measurements of the three dentists, neither the coronal plane nor the arch plane was statistically significant. (P > 0.05). CONCLUSION: This measurement method is simple, accurate and repeatable. It can be applied to measure alveolar bone height in the bone grafting area before and after maxillary sinus lift.

The 3D-Printing-Accelerated Design for a Biodegradable Respirator from Tree Leaves (TRespirator).

The unpredictable coronavirus pandemic (COVID-19) has led to a sudden and massive demand for face masks, leading to severe plastic pollution. Here, we propose a method for manufacturing biodegradable masks using high-precision 3D printing technology, called "TRespirator", mainly made of banana leaves and dental floss silk fibers. By adding plastic recycling waste appropriately, TRespirator can achieve similar protection and mechanical properties as N95 masks. In addition, microorganisms attracted during the degradation of plant fibers will accelerate the degradation of microplastics. This respirator provides a new idea for solving the global problem of plastic pollution of masks.

Effect of 45S5 bioactive glass on the microshear bond strength of dental fluorosis.

The aim of this in vitro study was to estimate the effect of the species concentration of 45S5 bioactive glass (BAG) used as pretreatment on the microshear bond strength (MSBS) of dental fluorosis (DF). Based on the Thylstrup and Fejerskov index, 80 teeth were randomly divided equally into four groups: TFI 0, sound dentin; TFI 1-3, mild fluorosis; TFI 4-5, moderate fluorosis; and TFI 6-9, severe fluorosis. Each group was randomized into five subgroups. After preparing the dentin hypersensitivity model of DF, the dentin was pretreated as follows, Subgroup 1: deionized water (Control group); Subgroup 2: 1% BAG; Subgroup 3: 5% BAG; Subgroup 4: 10% BAG, and Subgroup 5: 20% BAG. Stochastically one specimen was selected from each subgroup for scanning electron microscope and energy dispersive spectrometer analysis. After being made of resin-tooth bonding samples, the remains were in water bath at 37 °C for 24 hr. Subsequently, samples from each subgroup were randomly selected to test MSBS without aging, or after a thermocycle of 5,000 and 10,000 times, respectively. The fracture modes were analyzed. Compared with the group of 1% BAG and Control, the exposure area of tubules in 5%, 10%, and 20% BAG group had significant difference (p < .05). MSBS results indicated that there were significant differences between 10% BAG with other groups. The 20% BAG group showed the lowest MSBS among all groups. Pretreatment of 10% BAG solution may be conductive to enhance the bond strength of DF, while 20% BAG solution adversely.

Endogenous hydrogen peroxide in paraventricular nucleus mediates sympathetic activation and enhanced cardiac sympathetic afferent reflex in renovascular hypertensive rats.

An enhancement of the cardiac sympathetic afferent reflex (CSAR) contributes to sympathetic activation in renovascular hypertension. Angiotensin II in the paraventricular nucleus (PVN) augments the CSAR and increases sympathetic outflow and blood pressure. The present study aimed to determine whether endogenous hydrogen peroxide in the PVN mediated the enhanced CSAR, sympathetic activity and the effects of angiotensin II in the PVN in renovascular hypertension induced by the two-kidney, one-clip method (2K1C) in rats. At the end of the fourth week, the rats underwent sino-aortic and vagal denervation under general anaesthesia with urethane and α-chloralose. Renal sympathetic nerve activity (RSNA) and mean arterial pressure (MAP) were recorded. The CSAR was evaluated by the RSNA response to epicardial application of bradykinin. Microinjection of polyethylene glycol-catalase (PEG-CAT), an analogue of endogenous catalase, into the PVN decreased the RSNA and MAP and abolished the CSAR in both sham-operated and 2K1C rats. Microinjection into the PVN of the catalase inhibitor, aminotriazole, increased the RSNA and MAP and enhanced the CSAR. The effects of PEG-CAT or aminotriazole were greater in 2K1C rats than in sham-operated animals. The effects of angiotensin II in the PVN were abolished by pretreatment with PEG-CAT in both sham-operated and 2K1C rats; however, aminotriazole failed to potentiate the effects of angiotensin II. The catalase activity was decreased but the H(2)O(2) levels were increased in the PVN of 2K1C rats. These results indicate that endogenous H(2)O(2) in the PVN not only mediates the enhanced sympathetic activity and CSAR, but also the effects of angiotensin II in the PVN in renovascular hypertensive rats.

Tribological performance of organic molybdenum in the presence of organic friction modifier.

The tribological performance of organic molybdenum in the present of organic friction modifier was investigated in this study. Three types of organic friction modifiers were selected, which are Glycerol monooleate, Pentaerythritol and N,N-Dimethylhexadecylamine. The organic molybdenum are MoDTC, MoDDP and molybdenum amide. Friction coefficient and wear were studied in block-on-ring test rig with steel test specimens. Experimental results indicate the Pentaerythritol shows synergistic effect with MoDTC in wide range temperature, while increased the friction coefficient of molybdenum amide in high temperature. N,N-Dimethylhexadecylamine shows synergistic effect with molybdenum amide, while hindered the friction reduction performance of MoDTC in low temperature. The presence of Glycerol monooleate reduced friction coefficient of MoDTC in low temperature, while increased the friction coefficient of molybdenum amide in most situations. All the tested organic friction modifiers improved the friction reduction performance of MoDDP. Most of the tested organic friction modifiers reduced the wear of organic molybdenum. The PT shows the best anti-wear performance with MoDTC. The tribo-chemical products in test specimens lubricated with different lubricant formulas indicate that the presences of Pentaerythritol promotes the production of MoS2 in MoDTC. N,N-Dimethylhexadecylamine promotes the production of MoS2 in molybdenum amide. The side products of MoO1.6S1.6 and Cr/MoS2 of MoDDP in high temperature lead to high friction coefficient.

Treatment of oral lichen planus by surgical excision and acellular dermal matrix grafting: Eleven case reports and review of literature.

BACKGROUND: Oral lichen planus (OLP) is a chronic inflammatory disorder, and it can affect normal oral function. The conventional treatments for OLP are not always effective, and relapse easily occurs. Therefore, treatment of OLP is difficult and challenging. In this study, we evaluated over a long period the clinical efficacy of surgical excision and acellular dermal matrix (ADM) grafting in patients with refractory OLP. CASE SUMMARY: Eleven patients with refractory OLP underwent a standardized protocol of surgical excision and ADM grafting. The condition of the area of the grafted wound, the intraoperative maximum mouth opening, pain, and clinical healing were assessed at postoperative follow-up visits. All patients had a flat surgical area with similar mucosal tissue coverage and local scar formation. Patients had no irritation and pain in their mucous membranes when eating acidic and spicy food. All patients' mouth openings returned to normal within 2-6 mo after surgery. During follow-up, none of the patients had recurrence of OLP after surgery. The longest follow-up was 11 yr and the shortest was 6 mo, and none of the patients relapsed during follow-up. CONCLUSION: Surgical excision and ADM grafting could be an effective method to treat refractory OLP.

Screening of inhibitors against histone demethylation jumonji domain-containing protein 3 by capillary electrophoresis.

Jumonji domain-containing proteins (JMJDs) play an important role in the epigenetic regulation of gene expression. Aberrant regulation of histone modification has been observed in the progression of a variety of diseases, such as neurological disorders and cancer. Therefore, discovery of selective modulators of JMJDs is very attractive in new drug discovery. Herein, a simple capillary electrophoresis (CE) method was developed for screening of inhibitors against JMJD3. A known JMJD3 inhibitor GSK-J1, 5-carboxyfluorescein labeled substrate peptide with an amino acid sequence of KAPRKQLATKAARK(me3)SAPATGG (truncated from histone H3), as well as a small chemical library composed of 37 purified natural compounds and 30 natural extracts were used for method development and validation. The separation of substrate from its demethylated product was achieved by addition of polycation hexadimethrine bromide (HDB) in the running buffer. The enzyme activity was thus assayed accurately through separating the demethylated product from the substrate and then measuring the peak area of the product. The enzyme inhibition can be read out by comparing the peak area of the demethylated product obtained in the present of inhibitors and that of the negative control in the absence of any inhibitor. The merit of the method is proved by discovering two new JMJD3 inhibitors: salvianic acid A and puerarin 6''-O-xyloside.

Sandwich magnetically imprinted immunosensor for electrochemiluminescence ultrasensing diethylstilbestrol based on enhanced luminescence of Ru@SiO2 by CdTe@ZnS quantum dots.

A molecularly imprinted magnetic sensor with electroluminescent tags (MIP-ECL sensor) was developed for ultrasensing diethylstilbestrol (DES). A strategy is exploited to enhance ECL emission of the [Ru(bpy)3]2 +-tripropyl amine (TPrA) system by CdTe@ZnS quantum-dots (QDs) through energy transfer. Magnetically molecularly imprinted nanoparticles (MMIPs NPs) based on Fe3O4@SiO2 carriers are artificial, easily reproducible, and could replace easily inactivated first antibodies for capturing more DES molecules. Functionalized bio-conjugates of single antibody-CdTe@ZnS (Ab-CdTe@ZnS) are for the first time loaded on signal labels of Ru(bpy)32 +-doped silica nanocomposites (Ru@SiO2) for signal amplification. The final bio-conjugated signal probes are denoted as Ab-DES/CdTe@ZnS-Ru@SiO2. MMIPs beads that have captured antigens are bio-conjugated with antibody-labeled luminescent probes by specific immunoreactive reaction, and then the luminescent immunocomplex generates ECL signal on the magnetic electrode. The logarithm of ECL intensities depend linearly on the logarithm of DES concentrations in the range from 4.8 × 10- 4 to 36.0 nM with a detection limit of 0.025 pM. This novel assay is much more sensitive than other MIP sensors, and achieves lower cost and more enhanced stability than other immunosensors. The sensor is significantly potential and has been applied to DES detection in actual environment.

The effect of Lipoxin A4 on the interaction between macrophage and osteoblast: possible role in the treatment of aseptic loosening.

BACKGROUND: Aseptic loosening (AL) is the main problem of total joints replacement (TJR) by the implantation of permanently prosthetic components. In vitro and in vivo studies have clearly demonstrated that wear debris and its byproducts could trigger inflammation in the peri-implant tissue. Lipoxins (LXs) are endogenous eicosanoids synthesized locally from arachidonate acid (AA) at sites of inflammation and mediate pro-resolving activity. A number of studies have demonstrated the effect of LXA4 to counteract inflammation in different cell and animal models, but till now, no relative report about the role of LXs in progress or prevention of AL. METHODS: Murine RAW264.7 macrophage cell line and MC3T3-E1 osteoblasts (OB) cell line were purchased. Co-cultured model of these two cell lines was established. To explore the effect of exogenous Lipoxin A4 (LXA4) on polymethylmethacrylate (PMMA) induced inflammation, pro-inflammatory cytokines including TNF-alpha, IL-1beta, PGE2 and GM-CSF were measured by ELISA kits and bone resorption was quantified by measuring calcium release from 5-day-old mice calvaria in vitro. To determine further the endogenous effect of LXA4, cells were co-cultured and with or without 15-lipoxygease (15-LO) blocking by 15-LO siRNA. Both real-time PCR and western blotting were applied to confirm the inhibitory efficiency of 15-LO by siRNA. RESULTS: 0.1 mg/ml, 0.5 mg/ml and 1.0 mg/ml PMMA showed a time-dependent manner to trigger production of all the pro-inflammatory cytokines studied. Exogenous 0-100 nM LXA4 presented an inhibitory effect on both generation of above cytokines and PMMA stimulated calvarial bone resorption with a dose-dependent manner. LXA4 in supernatant from neither rest macrophages nor macrophages cultured alone exposing to PMMA was detectable. In co-cultured cells challenged by PMMA, LXA4 was increased significantly, while, this enhance could be partly inhibited by 15-LO siRNA. When LXA4 generation was blocked with 15-LO siRNA, the PMMA induced pro-inflammatory cytokines were elevated and bone resorption was accelerated. CONCLUSION: In the present study, we demonstrated that LXA4 had a favorable inhibitory effect on PMMA-induced inflammation in a macrophage and OB co-culture system.

PEGylated tumor cell membrane vesicles as a new vaccine platform for cancer immunotherapy.

Despite the promise and advantages of autologous cancer cell vaccination, it remains challenging to induce potent anti-tumor immune responses with traditional immunization strategies with whole tumor cell lysate. In this study, we sought to develop a simple and effective approach for therapeutic vaccination with autologous whole tumor cell lysate. Endogenous cell membranes harvested from cancer cells were formed into PEGylated nano-vesicles (PEG-NPs). PEG-NPs exhibited good serum stability in vitro and draining efficiency to local lymph nodes upon subcutaneous administration in vivo. Vaccination with PEG-NPs synthesized from murine melanoma cells elicited 3.7-fold greater antigen-specific cytotoxic CD8+ T lymphocyte responses, compared with standard vaccination with freeze-thawed lysate in tumor-bearing mice. Importantly, in combination with anti-programmed death-1 (αPD-1) IgG immunotherapy, PEG-NP vaccination induced 4.2-fold higher frequency of antigen-specific T cell responses (P < 0.0001) and mediated complete tumor regression in 63% of tumor-bearing animals (P < 0.01), compared with FT lysate + αPD-1 treatment that exhibited only 13% response rate. In addition, PEG-NPs + αPD-1 IgG combination immunotherapy protected all survivors against a subsequent tumor cell re-challenge. These results demonstrate a general strategy for eliciting anti-tumor immunity using endogenous cancer cell membranes formulated into stable vaccine nanoparticles.

[Drug release from electrospun simvastatin/polycaprolactone fibrous membranes].

PURPOSE: To prepare electrospun simvastatin/polycaprolactone(SMV/PCL) membrane scaffolds and to evaluate the release properties of this formulation. METHODS: Electrospun SMV/PCL membrane scaffolds were prepared as the experimental group, and electrospun PCL membrane as the control group. The morphology and characteristics of membrane surface were determined by scanning electron microscopic(SEM) and X-ray diffraction(XRD). The release profile of SMV was determined using an ultraviolet-visible spectrometer. The data were analyzed statistically with SPSS 17.0 software package. RESULTS: SEM and XRD indicated that SMV/PCL nanofibers were successfully electrospun and SMV was encapsulated into the fibers. In vitro drug release studies showed that simultaneous SMV release, being nearly linear with time, was achieved and sustained SMV release was prolonged to 28 days. CONCLUSIONS: Electrospun SMV/PCL nanofiber membranes demonstrate sustained drug release properties, suggesting their potential applicability as prospective scaffolds in periodontal regeneration.