Nicotine aggravates pancreatic fibrosis in mice with chronic pancreatitis via mitochondrial calcium uniporter.

INTRODUCTION: This study aimed to investigate the effects of nicotine on the activation of pancreatic stellate cells (PSCs) and pancreatic fibrosis in chronic pancreatitis (CP), along with its underlying molecular mechanisms. METHODS: This was an in vivo and in vitro study. In vitro, PSCs were cultured to study the effects of nicotine on their activation and oxidative stress. Transcriptome sequencing was performed to identify potential signaling pathways involved in nicotine action. And the impact of nicotine on mitochondrial Ca2+ levels and Ca2+ transport-related proteins in PSCs was analyzed. The changes in nicotine effects were observed after the knockdown of the mitochondrial calcium uniporter (MCU) in PSCs. In vivo experiments were conducted using a mouse model of CP to assess the effects of nicotine on pancreatic fibrosis and oxidative stress in mice. The alterations in nicotine effects were observed after treatment with the MCU inhibitor Ru360. RESULTS: In vitro experiments demonstrated that nicotine promoted PSCs activation, characterized by increased cell proliferation, elevated α-SMA and collagen expression. Nicotine also increased the production of reactive oxygen species (ROS) and cellular malondialdehyde (MDA), exacerbating oxidative stress damage. Transcriptome sequencing revealed that nicotine may exert its effects through the calcium signaling pathway, and it was verified that nicotine elevated mitochondrial Ca2+ levels and upregulated MCU expression. Knockdown of MCU reversed the effects of nicotine on mitochondrial calcium homeostasis, improved mitochondrial oxidative stress damage and structural dysfunction, thereby alleviating the activation of PSCs. In vivo validation experiments showed that nicotine significantly aggravated pancreatic fibrosis in CP mice, promoted PSCs activation, exacerbated pancreatic tissue oxidative stress, and increased MCU expression. However, treatment with Ru360 significantly mitigated these effects. CONCLUSIONS: This study confirms that nicotine upregulates the expression of MCU, leading to mitochondrial calcium overload and exacerbating oxidative stress in PSCs, and ultimately promoting PSCs activation and exacerbating pancreatic fibrosis in CP.

Mitochondria oxidative stress mediated nicotine-promoted activation of pancreatic stellate cells by regulating mitochondrial dynamics.

Nicotine, one of the main ingredients of cigarettes, promotes activation of pancreatic stellate cells(PSCs) and exacerbates pancreatic fibrosis in previous studies. Here we focus on the inner relationship between mitochondrial oxidative stress and mitochondrial dynamics to explore the possible mechanism. Primary human PSCs were stimulated by nicotine. The effect of nicotine on oxidative stress and mitochondrial dynamics was analyzed by reactive oxygen species (ROS) assay, quantitative real-time PCR, and western blotting. Mitochondrial morphology was observed. Antioxidant and small interfering RNA transfection were applied to explore the interrelationship between oxidative stress and mitochondrial dynamics, as well as its effect on PSCs activation. Nicotine exposure significantly increased Intracellular and mitochondrial ROS of hPSCs and promoted mitochondrial fission by upregulating dynamin-related protein 1(DRP1). Knockdown Drp1 reversed mitochondrial fragmentation and hPSCs activation that promoted by nicotine, but fail to alleviate oxidative stress. A mitochondrial-targeted antioxidant could reverse all the above changes. Our finding suggests that mitochondria oxidative stress mediated nicotine-promoted activation of PSCs by inducing Drp1-mediated mitochondrial fission, provides a new perspective on the possible mechanism by which nicotine affects PSCs, and reveals a potential therapeutic strategy.

Nicotine facilitates pancreatic fibrosis by promoting activation of pancreatic stellate cells via α7nAChR-mediated JAK2/STAT3 signaling pathway in rats.

AIM: Smoking has been considered as a risk factor of chronic pancreatitis (CP), but the potential mechanism is still unknown. The major pathological feature of CP is pancreatic fibrosis, whose major functional cells are pancreatic stellate cells (PSCs). Nicotine is the major component of cigarette smoke, our recent study suggested that nicotine has the potential to facilitate pancreatic fibrosis in CP. This study was aimed to analyze the function and mechanism of nicotine on PSCs and pancreatic fibrosis in rats. MATERIALS AND METHODS: In vivo, a rat CP model was induced by intraperitoneal injection of 20 % L-arginine hydrochloride (200 mg/100 g) at 1 h intervals twice per week, nicotine was injected subcutaneously at a dose of 1 mg/kg body weight per day. After four weeks, the pancreatic tissue was collected for H&E, Masson and immunohistochemical staining. In vitro, primary rPSCs were isolated from rats and treated with nicotine (0.1 µM and 1 µM). The proliferation、apoptosis、α-SMA expression、extracellular matrix (ECM) metabolism and α7nAChR-mediated JAK2/STAT3 signaling pathway of rPSCs were detected by CCK-8 assay、flow cytometry、real-time Q-PCR and western blotting analysis. The α7nAChR antagonist α-bungarotoxin (α-BTX) was used to perform inhibition experiments. KEY FINDINGS: Nicotine increased pancreatic damage, collagen deposition and activation of PSCs in the CP rat model. In rPSCs, the proliferation, α-SMA expression and ECM formation were significantly promoted by nicotine in a dose-dependent manner. Meanwhile, the apoptosis of rPSCs was significantly reduced after nicotine treatment. Moreover, nicotine also activated the α7nAChR-mediated JAK2/STAT3 signaling pathway in rPSCs. These effects of nicotine on rPSCs were blocked by α-BTX. SIGNIFICANCE: Our finding in this research suggests that nicotine facilitates pancreatic fibrosis by promoting activation of pancreatic stellate cells via α7nAChR-mediated JAK2/STAT3 signaling pathway in rats, partly revealing the mechanism of smoking on chronic pancreatitis.

Tetrafunctional Epoxy Resin-Based Buoyancy Materials: Curing Kinetics and Properties.

In order to synthesize a new kind of buoyancy material with high-strength, low-density and low-water-absorption and to study the curing reaction of tetraglycidylamine epoxy resin with an aromatic amine curing agent, the non-isothermal differential scanning calorimeter (DSC) method is used to calculate the curing kinetics parameters of N,N,N',N'-tetraepoxypropyl-4,4'-diaminodiphenylmethane epoxy resin (AG-80) and the m-xylylenediamine (m-XDA) curing process. Further, buoyancy materials with different volume fractions of hollow glass microsphere (HGM) compounded with a AG-80 epoxy resin matrix were prepared and characterized. The curing kinetics calculation results show that, for the curing reaction of the AG-80/m-XDA system, the apparent activation energy increases with the conversion rates increasing and the reaction model is the Jander equation (three-dimensional diffusion, 3D, n = 1/2). The experimental results show that the density, compressive strength, saturated water absorption and water absorption rate of the composite with 55 v % HGM are 0.668 g·cm-3, 107.07 MPa, 0.17% and 0.025 h-1/2, respectively. This kind of composite can probably be used as a deep-sea buoyancy material.

Nicotine promotes activation of human pancreatic stellate cells through inducing autophagy via α7nAChR-mediated JAK2/STAT3 signaling pathway.

AIM: Pancreatic stellate cells (PSCs) are the main functional cells leading to pancreatic fibrosis. Nicotine is widely considered as an independent risk factor of pancreatic fibrosis, but the mechanism is still unclear. Our study was aimed to explore the effects of nicotine on human pancreatic stellate cells (hPSCs) and involved pathways. MATERIALS AND METHODS: Primary human PSCs were cultured and treated with nicotine (0.1 µM and 1 µM). The proliferation, apoptosis, α-SMA expression, extracellular matrix metabolism and autophagy of hPSCs were detected by CCK-8 assay, flow cytometry, real-time PCR and Western blotting analysis. The α7nAChR-mediated JAK2/STAT3 signaling pathway was also examined, and an α7nAChR antagonist α-bungarotoxin (α-BTX) was used to perform inhibition experiments. KEY FINDINGS: The proliferation, α-SMA expression and autophagy of hPSCs were significantly promoted by 1 µM nicotine. Meanwhile, the apoptosis of hPSCs was significantly reduced. The extracellular matrix metabolism of hPSCs was also regulated by nicotine. Moreover, the α7nAChR-mediated JAK2/STAT3 signaling pathway was activated by nicotine, this pathway and effects of nicotine can be blocked by α-BTX. SIGNIFICANCE: Our finding suggests that nicotine can promote activation of human pancreatic stellate cells (hPSCs) through inducing autophagy via α7nAChR-mediated JAK2/STAT3 signaling pathway, providing a new insight into the mechanisms by which nicotine affects pancreatic fibrosis.

Curing and Characteristics of N,N,N',N'-Tetraepoxypropyl-4,4'-Diaminodiphenylmethane Epoxy Resin-Based Buoyancy Material.

Buoyancy material is a type of low-density and high-strength composite material which can provide sufficient buoyancy with deep submersibles. A new buoyancy material with N,N,N',N'-tetraepoxypropyl-4,4'-diaminodiphenylmethane epoxy resin (AG-80) and m-xylylenediamine (m-XDA) curing agent as matrix and hollow glass microsphere (HGM) as the filler is prepared. The temperature and time of the curing process were determined by the calculations of thermal analysis kinetics (TAK) through differential scanning calorimetry (DSC) analysis. The results show that the better mass ratio of AG-80 with m-XDA is 100/26. Combined TAK calculations and experimental results lead to the following curing process: pre-curing at 75 °C for 2 h, curing at 90 °C for 2 h, and post-curing at 100 °C for 2 h. The bulk density, compressive strength, and saturated water absorption of AG-80 epoxy resin-based buoyancy material were 0.729 g/cm3, 108.78 MPa, and 1.23%, respectively. Moreover, this type of buoyancy material can resist the temperature of 250 °C.

Bivariate optimization of orthodontic mini-implant thread height and pitch.

PURPOSE: Mini-implants have been used as anchorage for years, but failure is common in clinical practice. Mini-implant design is a critical factor affecting its stability. The aim of this study was to evaluate the effect of continuous and simultaneous variations of thread height and pitch on the biomechanical properties of an orthodontic mini-implant. METHOD: A 3D finite element model, composed of a posterior maxilla section and an orthodontic mini-implant, was created. Mini-implant thread height ranged from 0.10 to 0.40 mm, and thread pitch ranged from 0.50 to 2.00 mm. Effects of the implant thread height and pitch on the maximum Von Mises stresses in maxilla and mini-implant, as well as maximum displacements in the mini-implant, were evaluated by a finite element method. Bivariate analysis was used to determine the optimal range of thread height and pitch. RESULTS: Variation of thread height and pitch decreased the maximum Von Mises stresses in cortical bone, cancellous bone and mini-implant by 54.9, 78.4 and 23.6 %, respectively. The maximum displacement in the mini-implant decreased by 21.8 %. CONCLUSION: Maxillary stress and mini-implant stability were influenced by mini-implant thread height and pitch. Increased thread height with a thread pitch of 1.20 mm was better for orthodontic mini-implant in the maxillary posterior region. Thread height played a more significant role than the thread pitch in reducing maxillary stress and enhancing orthodontic mini-implant stability.

[Clinical study of repair the defect of immediate implant by acellular dermal matrix].

OBJECTIVE: To evaluate the clinical effect of acellular dermal matrix allograft in repairing the oral mucosal defect of immediate implant. METHODS: 51 cases underwent immediate implant surgery for 67 implants right after the teeth or roots extracted. The mucosal defect of implant areas were repaired by acellular dermal matrix. The basal membrane side of acellular dermal matrix was exposed to oral cavity, and another side was attached to the implant and alveolar crest surface. It was intercalated between mucosal flap and alveolar and fixed by iodoform pack or base plate. To understand condition of wound healing the patients were followed up from 4 months to 6 months after operation. The acellular dermal matrix closed wound and histological changes were observed. The implant was followed up months to 4 years. RESULTS: The wounds were completely healed in 54 implant areas, partially healed in 11 implant areas, not healed 2 implant areas. histological examination wasn't differentiation between mucous epithelium and graft epithelium. None of 67 implants showed deterioration in the follow-up of one year. It was no obvious sign of immune rejection. CONCLUSION: The clinical result of acellular dermal matrix in repair the mucosal defect of immediate implant is good, the advantages are not to affect the integration bone with implant, less operation trauma, good esthetics results.