Phytoremediation performance of mixed aquatic plantings and their rhizosphere microbial community in pilot-scale constructed wetlands.

Phytoremediation is a low-cost, environmentally friendly, and sustainable technology that can utilize vegetation and microorganisms to avoid eutrophication and purifying water environment. The ability of five different living aquatic plants of nitrogen (N), phosphorus (P), and chemical oxygen demand (CODcr) removal were investigated in pilot scale constructed wetlands (CWs). Aquatic plant mixes significantly improved CODcr removal and plant tissue uptake of nitrogen and phosphorus. The wetland performance of mixed plantings was also influenced by the specific species. When assessing pollutant removal in CWs, PNM performed better within mixtures, a possible synergistic effect, while TNV performed poorly, a possible antagonist effect. The nutrient uptake within plant tissues by mixed plants was always a synergistic effect. Aquatic plant mixes significantly increased the rhizosphere microbial diversity and promoted the growth of functional heterotrophic denitrifying flora.

Colloid Mill-Assisted Ultrasonic-Fractional Centrifugal Purification of Low-Grade Attapulgite and Its Modification for Adsorption of Congo Red.

Attapulgite (APT) is widely used in wastewater treatment due to its exceptional adsorption and colloidal properties, as well as its cost-effectiveness and eco-friendliness. However, low-grade APT generally limits its performance. Here, a colloid mill-assisted ultrasonic-fractional centrifugal purification method was developed to refine low-grade APT. This process successfully separated and removed impurity minerals such as quartz and dolomite from the raw ore, resulting in a refined APT purity increase from 16.9% to 60% with a specific surface area of 135.5 m2∙g-1. Further modifying of the refined APT was carried out through the hydrothermal method using varying dosages of cetyltrimethylammonium chloride (CTAC), resulting in the production of four different APT adsorbents denoted as QAPT-n (n = CTAC mole number) ranging from 0.5 to 5 mmol. Using Congo red (CR) as the target pollutant, the QAPT-5 sample exhibited the best adsorption capacity with the maximum quantity of 1652.2 mg∙g-1 in a neutral solution at 30 °C due to the highest surface charge (zeta potential = 8.25 mV). Moreover, the QAPT-5 pellets (~2.0 g adsorbent) shaped by the alginate-assisted molding method removed more than 96% of 200 mL aqueous solution containing 200 mg∙L-1 CR and maintained this efficiency in 10 adsorption-elution cycles, which exhibited the promising practical application.

Correlation analysis and clinical significance of changes in upper thoracic vertebra tilt and clavicle angle pre- and post-operation.

The imbalance of the lateral shoulder is reflected by the clavicle angle (CA) in radiology. It remains unclear how to achieve postoperative lateral shoulder balance (LSB) after spinal deformity correction surgery. A retrospective analysis was conducted on AIS patients who underwent surgery by the same spine surgeon at our hospital from 2016 to 2020. A total of 110 patients with spinal deformity were included in the study to verify the correlation between the T1-T5 tilt angle and CA before and after surgery, as well as the relation-ship between the change in T1-T5 tilt angle before and after surgery and the change in CA before and after surgery. By comparing the correlation coefficients, it was found that there may not be a direct relationship between the pre- and postoperative tilt angles of T1-5 and CA, but their changes were closely related to the changes in CA. The change in T1 tilt angle after orthopaedic surgery was significantly correlated with the change in CA, with a correlation coefficient of 0.976, indicating a close relationship between T1 and the clavicle. As the vertebrae moved down, the correlation gradually decreased. In summary, this study suggests that there is a close relationship between T1-T5 and the clavicle and that the change in T1 tilt angle after spinal scoliosis correction surgery is significantly correlated with CA, which decreases as the vertebra moves down.

Calpain Inhibitor Calpeptin Improves Pancreatic Fibrosis in Mice with Chronic Pancreatitis by Inhibiting the Activation of Pancreatic Stellate Cells.

BACKGROUND: Pancreatic fibrosis is a hallmark feature of chronic pancreatitis (CP), resulting in persistent damage to the pancreas. The sustained activation of pancreatic stellate cells (PSCs) plays a pivotal role in the progression of pancreatic fibrosis and is a major source of extracellular matrix (ECM) deposition during pancreatic injury. METHODS: Calpain is a calcium-independent lysosomal neutral cysteine endopeptidase and was found to be correlated to various fibrotic diseases. Studies have revealed that calpeptin, a calpain inhibitor, can improve the fibrosis process of multiple organs. This study investigated the effect of the calpain inhibitor, calpeptin, on fibrosis in experimental CP and activation of cultured PSCs in mice. CP was induced in mice by repeated injections of cerulein for four weeks in vivo, and the activation process of mouse PSCs was isolated and cultured in vitro. Then, the inhibitory effect of calpeptin on pancreatic fibrosis was confirmed based on the histological damage of CP, the expression of α-smooth muscle actin (α-SMA) and collagen-Iα1(Col1α1), and the decrease in mRNA levels of calpain-1 and calpain-2. RESULTS: In addition, it was revealed that calpeptin can inhibit the activation process of PSCs and induce significant PSCs apoptosis by downregulating the expression of calpain-1, calpain-2 and TGF-ß1, and the expression and phosphorylation of smad3 in vitro. CONCLUSION: These results suggest that the calpain inhibitor, calpeptin, plays a key role in the regulation of PSC activation by inhibiting the TGF-ß1/smad3 signaling pathway, which supports the potential of calpeptin as an inhibitor of pancreatic fibrosis in mice by interfering with calpain.

Boosting electrocatalytic performance via electronic structure regulation for acidic oxygen evolution.

High-purity hydrogen produced by water electrolysis has become a sustainable energy carrier. Due to the corrosive environments and strong oxidizing working conditions, the main challenge faced by acidic water oxidation is the decrease in the activity and stability of anodic electrocatalysts. To address this issue, efficient strategies have been developed to design electrocatalysts toward acidic OER with excellent intrinsic performance. Electronic structure modification achieved through defect engineering, doping, alloying, atomic arrangement, surface reconstruction, and constructing metal-support interactions provides an effective means to boost OER. Based on introducing OER mechanism commonly present in acidic environments, this review comprehensively summarizes the effective strategies for regulating the electronic structure to boost the activity and stability of catalytic materials. Finally, several promising research directions are discussed to inspire the design and synthesis of high-performance acidic OER electrocatalysts.

Effects of Foliar Selenium Application on Oxidative Damage and Photosynthetic Properties of Greenhouse Tomato under Drought Stress.

Both drought stress and exogenous selenium (Se) cause changes in plant physiological characteristics, which are key factors affecting crop yield. Although Se is known to be drought-resistant for crops, its internal physiological regulatory mechanisms are not clear. This study analyzed the effects of selenium application (SeA) on antioxidant enzyme activities, osmoregulatory substance contents, and photosynthetic characteristics of greenhouse tomatoes under drought stress and related physiological mechanisms. The results showed that drought stress induced oxidative damage in cells and significantly increased the content of the membrane lipidation product malondialdehyde (MDA) and the osmoregulatory substance proline (p < 0.001) compared with the adequate water supply. The proline content of severe drought stress (W1) was 9.7 times higher than that of the adequate water supply (W3), and foliar SeA increased glutathione peroxidase (GSH-PX) activity, and SeA induced different enzymatic reactions in cells under different drought stresses; catalase (CAT) was induced under severe drought stress (p < 0.01) and was significantly increased by 32.1% compared with the clear water control, CAT. Peroxidase (POD) was induced under adequate water supply conditions (p < 0.01), which was significantly increased by 15.2%, and SeA attenuated cell membrane lipidation, which reduced MDA content by an average of 21.5% compared with the clear water control, and also promoted photosynthesis in the crop. Meanwhile, through the entropy weighting method analysis (TOPSIS) of the indexes, the highest comprehensive evaluation score was obtained for the S5W3, followed by the S2.5W3 treatment. Therefore, this study emphasized the importance of SeA to reduce oxidative damage and enhance photosynthesis under drought stress.

Qing Xia Jie Yi Formula granules alleviated acute pancreatitis through inhibition of M1 macrophage polarization by suppressing glycolysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Herbal formulas from Traditional Chinese Medicine are common and well-established practice for treating acute pancreatitis (AP) patients. However, little is known about their bioactive ingredients and mechanisms, such as their targets and pathways to inhibit inflammation. AIM OF THE STUDY: This study aimed to evaluate the effect of Qing Xia Jie Yi Formula (QXJYF) granules on AP and discuss the molecular mechanisms involved. MATERIALS AND METHODS: Major compounds in QXJYF granules were identified using UPLC-quadrupole-Orbitrap mass spectrometry (UPLC-Q-Orbitrap MS). The effect of QXJYF granules on experimental AP models both in vitro and in vivo, and detailed mechanisms were clarified. Two AP models were induced in mice by intraperitoneally injections of caerulein or L-arginine, and QXJYF granules were used to treat AP mice in vivo. Histological evaluation of pancreas and lung, serum amylase and lipase levels, serum inflammatory cytokines, inflammatory cell infiltration and macrophage phenotype were assessed. Bone marrow derived macrophages (BMDMs) were cultured and treated with QXJYF granules in vitro. BMDM phenotype and glycolysis levels were measured. Lastly, clinical effect of QXJYF granules on AP patients was verified. Predicted severe AP (pSAP) patients eligible for inclusion were assessed for enrollment. RESULTS: Nine major compounds were identified in QXJYF granules. Data showed that QXJYF granules significantly alleviated AP severity both in caerulein and L-arginine-induced AP models in vivo, pancreatic injury and inflammatory cell infiltration, systematic inflammation, lung injury and inflammatory cell infiltration were all improved after QXJYF treatment. QXJYF granules significantly reduced M1 macrophages during AP both in vivo and in vitro; besides, the mRNA expression levels of M1 genes such as inos, Tnfα, Il1ß and Il6 were significantly lower after QXJYF treatment in M1 macrophages. Mechanistically, we found that HK2, PFKFB3, PKM, LDHα levels were increased in M1 macrophages, but significantly decreased after QXJYF treatment. Clinical data indicated that QXJYF granules could significantly reduce CRP levels and shorten the duration of organ failure, thereby reducing the incidence of SAP and preventing pSAP patients from progressing to SAP. CONCLUSION: QXJYF granules alleviated AP through the inhibition of M1 macrophage polarization by suppressing glycolysis.

Effects of Post-Anthesis Irrigation on the Activity of Starch Synthesis-Related Enzymes and Wheat Grain Quality under Different Nitrogen Conditions.

To develop optimal management strategies for water and nitrogen fertilizer application in winter wheat cultivation, we conducted a potted experiment to investigate the effects of different irrigation levels and nitrogen fertilizer treatments on the activity of starch synthesis-related enzymes and the grain quality of winter wheat. The potted experiment consisted of three irrigation levels, with the lower limits set at 50-55% (I0), 60-65% (I1), and 70-75% (I2) of the field capacity. In addition, four levels of nitrogen fertilizer were applied, denoted as N0 (0 kg N hm-2), N1 (120 kg N hm-2), N2 (240 kg N hm-2), and N3 (300 kg N hm-2), respectively. The results revealed the significant impacts of irrigation and nitrogen treatments on the activities of key starch-related enzymes, including adenosine diphosphoglucose pyrophosphrylase (ADPG-PPase), soluble starch synthase (SSS), granule-bound starch synthase (GBSS), and starch branching enzymes (SBE) in wheat grains. These treatments also influenced the starch content, amylopectin content, and, ultimately, wheat yield. In summary, our findings suggest that maintaining irrigation at a lower limit of 60% to 65% of the field capacity and applying nitrogen fertilizer at a rate of 240 kg hm-2 is beneficial for achieving both high yield and high quality in winter wheat cultivation.

Foliar Application of Melatonin Positively Affects the Physio-Biochemical Characteristics of Cotton (Gossypium hirsutum L.) under the Combined Effects of Low Temperature and Salinity Stress.

Low temperature and soil salinization during cotton sowing and seedling adversely affect cotton productivity. Exogenous melatonin (MT) can alleviate the damage caused to plants under non-biological stress; thus, applying MT is a means to improve the growth condition of crops under stress. However, achieving this goal requires a thorough understanding of the physiological regulatory mechanisms of MT on cotton seedlings under low temperature and salinity stress. This study could bring new knowledge on physio-biochemical mechanisms that improve the tolerance of cotton seedlings to combined effects of low temperature and salt stress using an exogenous foliar application of MT. The phytotron experiment comprised two temperature levels of cold stress and control and five MT treatments of 0, 50, 100, 150, and 200 µM and two salinity levels of 0 and 150 mM NaCl. Compared with the control treatments (non-salinity stress under cold stress and control), the combined stress of salt and low temperature reduced cotton seedlings' biomass and net photosynthetic rate (Pn), aggravated the membrane damage, reduced the potassium (K+) content, and increased the sodium (Na+) accumulation in the leaves and roots. Under NaCl stress, exogenously sprayed 50-150 µM MT increased the biomass and gas exchange parameters of cotton seedlings under salt and low temperature combined with salt stress, reduced the degree of membrane damage, and regulated the antioxidant enzyme, ion homeostasis, transport, and absorption of cotton seedlings. The pairwise correlation analysis of each parameter using MT shows that the parameters with higher correlation with MT at cold stress are mainly malondialdehyde (MDA), peroxidase (POD), and catalase (CAT). The highest correlation coefficient at 25 °C is observed between the K+ and Na+ content in cotton seedlings. The conclusion indicates that under salt and low-temperature stress conditions, exogenous application of MT primarily regulates the levels of Pn, superoxide dismutase (SOD), andPOD in cotton seedlings, reduces Na+ and MDA content, alleviates damage to cotton seedlings. Moreover, the most significant effect was observed when an exogenous application of 50-150 µM of MT was administered under these conditions. The current study's findings could serve as a scientific foundation for salinity and low-temperature stress alleviation during the seedling stage of cotton growth.

Suppression of Wnt/ß-catenin Signaling in PDAC via METTL16-mediated N6-methyladenosine Modification of DVL2.

Pancreatic cancer is a formidable cause of cancer-related deaths worldwide and has witnessed a more than twofold increase in incidence over the last 25 years. The most frequently occurring form of pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC), accounting for the majority of pancreatic cancer cases. N6-methyladenosine (m6A), the most abundant transcript modification, has been implicated in the pathogenesis of numerous human cancers, including pancreatic cancer. Despite this, the functional role of methyltransferase-like 16 (METTL16), a critical m6A methyltransferase, in PDAC remains elusive. In this study, we demonstrate that METTL16 expression is significantly diminished in PDAC, rendering it a promising prognostic indicator. Strikingly, both in vitro and in vivo assays revealed accelerated metastasis and invasion of PDAC cells upon METTL16 knockdown, while overexpression of METTL16 exerted an opposite effect. Mechanistically, METTL16 regulates DVL2 expression by suppressing its translation via m6A modification, thereby regulating Wnt/ß-catenin signaling., Our results unveil the downregulation of METTL16 as a concomitant increase in DVL2 levels via m6A modification promoting the progression of PDAC. Thus, we propose METTL16 as a novel therapeutic candidate for targeted PDAC treatment.

Supraspinous ligament arc tangent guided freehand thoracic pedicle screw insertion technique: high parallelism between screws and upper endplate.

Research objective: To propose a technique for placing pedicle screws in the thoracic spine using the Supraspinous ligament Arc Tangent (SLAT) as a guide to increase the safety and stability of screw placement. Content and methods: A retrospective analysis of postoperative anteroposterior and lateral x-ray images was performed for 118 patients with thoracic spine diseases who received conventional freehand technique from January 2016 to May 2020 and SLAT-guided technique since June 2020 to present. The diagnoses included thoracic spinal stenosis, deformity, fractures, infections, and tumors. The angle between the screw and the upper endplate was categorized as grade 1 (0°-5°), grade 2 (5°-10°), and grade 3 (>10°). Three surgeons with more than 10 years of experience in spinal surgery measured the angle between the screw and the upper endplate in the lateral view. Chi-square test was used for statistical analysis, and p < 0.05 was considered statistically significant. Results: A total of 1315 pedicle screws were placed from T1 to T12 in all patients. In the conventional freehand technique group, 549 screws were grade 1, 35 screws were grade 2, and 23 screws were grade 3. In the SLAT-guided freehand technique group, 685 screws were grade 1, 15 screws were grade 2, and 8 screws were grade 3. The data of each group was p < 0.05 by Chi-squared test, which was statistically significant, indicating that the SLAT-guided freehand technique resulted in a higher rate of parallelism between the screws and the upper endplate. All patients underwent intraoperative neurophysiological monitoring, immediate postoperative neurological examination, postoperative x-ray examination, and assess the eventual recovery. The screws were safe and stable, and no complications related to pedicle screw placement were found. Conclusion: The SLAT-guided freehand technique for placing pedicle screws in the thoracic spine can achieve a higher rate of screw-upper endplate parallelism, making screw placement safer and more accurate. Our method provides a convenient and reliable technique for most spinal surgeons, allowing for increased accuracy and safety with less fluoroscopic guidance.

Effects of Single and Combined Drought and Salinity Stress on the Root Morphological Characteristics and Root Hydraulic Conductivity of Different Winter Wheat Varieties.

Water shortages and crop responses to drought and salt stress are related to the efficient use of water resources and are closely related to food security. In addition, PEG or NaCl stress alone affect the root hydraulic conductivity (Lpr). However, the effects of combined PEG and NaCl stress on Lpr and the differences among wheat varieties are unknown. We investigated the effects of combined PEG and NaCl stress on the root parameters, nitrogen (N) and carbon content, antioxidant enzymes, osmotic adjustment, changes in sodium and potassium, and root hydraulic conductivity of Yannong 1212, Heng 4399, and Xinmai 19. PEG and NaCl stress appreciably decreased the root length (RL), root surface area (RS), root volume (RV), K+ and N content in shoots and roots, and Lpr of the three wheat varieties, while the antioxidant enzyme activity, malondialdehyde (MDA), osmotic adjustment, nonstructural carbon and Na+ content in shoots and roots, etc., remarkably remained increased. Furthermore, the root hydraulic conductivity had the greatest positive association with traits such as RL, RS, and N and K+ content in the shoots of the three wheat varieties. Moreover, the RL/RS directly and actively determined the Lpr, and it had an extremely positive effect on the N content in the shoots of wheat seedlings. Collectively, most of the root characteristics in the wheat seedlings decreased under stress conditions, resulting in a reduction in Lpr. As a result, the ability to transport nutrients-especially N-from the roots to the shoots was affected. Therefore, our study provides a novel insight into the physiological mechanisms of Lpr.

Risk factors associated with left atrial appendage thrombosis in patients with non-valvular atrial fibrillation by transesophageal echocardiography.

PURPOSE: This study investigated possible mechanism of left atrial appendage (LAA) thrombosis and constructed a model to evaluate the future risk of LAA thrombosis and spontaneous echo contrast (SEC) in non-valvular atrial fibrillation (NVAF) patients. METHODS: This retrospective study included 2591 patients diagnosed with NVAF. Patients were divided based on the presence of transesophageal echocardiography (TEE) into a thrombus group, SEC group, and control group. General, biochemical, and echocardiography data of the three groups were analyzed. The variables independently associated with LAA thrombosis and SEC were determined by the logistic regression analysis. A nomogram was constituted based on the regression analysis and the discriminatory ability was analyzed by receiver operating characteristic (ROC) curve. RESULTS: LAA thrombosis and SEC were present in 110 (4.2%) patients and 103 (3.9%) patients, respectively. AF type (OR = 1.857), previous stroke (OR = 1.924), fibrinogen (OR = 1.636), diameters of the left atria (OR = 1.094), left ventricular ejection fraction (OR = 0.938), and LAA maximum caliber (OR = 1.238) resulted as independent risk factors for LAA thrombosis and SEC. The area under curve of the nomogram established by multivariate logistic regression was 0.824. Conclusions; Through the study, 6 independent risk factors related to the LAA thrombosis and SEC were found, and an effective nomogram was constructed to predict the LAA thrombosis and SEC in NVAF patients.

Acute Spinal Epidural Abscess of the Cervical Spine Caused by Streptococcus constellatus Leads to Paraplegia in an Adult: A Case Report.

The incidence of a spinal epidural abscess (SEA), which can cause serious neurological complications, is low; however, the incidence of SEA caused by Streptococcus is even lower, most of which are reported in the thoracolumbar spine and lumbosacral segment. We reported a case of cervical SEA caused by Streptococcus constellatus infection, resulting in paralysis of the patient. The acute onset of SEA in a 44-year-old male led to decreased upper limb muscle strength, lower limb paralysis, and loss of bowel and bladder function, and imaging and blood tests suggested pyogenic spondylitis. Emergency decompression surgery and antibiotic therapy were given, the patient gradually recovered, and the muscle strength of the lower limbs gradually improved. This case report suggests the importance of early decompressive surgery and effective antibiotic therapy.

Irrigation modulates the effect of increasing temperatures under climate change on cotton production of drip irrigation under plastic film mulching in southern Xinjiang.

Introduction: Warming and drought brought about by climate change seriously harm sustainable agricultural production in southern Xinjiang. It is still unclear how irrigation can improve the ability of crops to cope with climate change. Methods: Therefore, in this study, we calibrated and validated the AquaCrop model using data collected in cotton production from 2017 to 2018. The model effectively simulated the growth, biomass, and yield of cotton plants at the experimental site under different warming and irrigation conditions. The meteorological data collected from 1987 to 2016 were used in a simulation to predict cotton production under 3 temperature scenarios (temperature increased by 0°C, 1°C, and 2°C) and 6 levels of irrigation (198, 264, 330, 396, 495, and 594 mm) to explain the modulating effect of plastic film mulching-coupled drip irrigation on cotton production in terms of increasing temperatures under climate change in southern Xinjiang. Results and discussion: Model prediction showed that an increase in temperature reduced cotton yield under a low irrigation level, while an increase in irrigation mitigated the impact of climate change on cotton yield. An increase of 1°C did not significantly reduce cotton yield at 198-330 mm of irrigation. Under a 2°C increase, 396-594 mm of irrigation was required to ensure plant growth and yield formation. Both aboveground biomass and yield increased with the rise in the irrigation level at the same temperature. High water use efficiency was achieved at 495 mm of irrigation without significant yield loss. Therefore, in the low-temperature scenario, it can be preferentially considered to achieve sustainable water use through water management, while in the high-temperature scenario innovative agricultural measures are required to avoid yield loss. Optimizing irrigation strategies can reduce warming-induced damage to crops under climate change.

Effect of Plant Growth Regulators on Cotton Seedling Root Growth Parameters and Enzyme Activity.

It is well known that the survival rate of cotton seedlings is low, and the growth and development status at this stage is crucial to improve productivity. Plant hormones are important factors that promote the growth and development of cotton seedlings. Growth regulators have the same function as plant hormones. The purpose of this research is to explore the effects of different concentrations of growth regulators on cotton root morphological parameters and enzyme activities, and to find suitable plant growth regulators and their optimal concentrations to improve the growth of the cotton seedling root system. Three cotton varieties, "Zhongmian 619" (Z619), "Xinluzao 27" (Z27), and "Xinluzao 39" (Z39), and three growth regulators, gibberellin (GA3), salicylic acid (SA), and paclobutrazol (PP333), at three concentrations were used in our experiment. In Z619 and Z27, 0.050 mg/L GA3 significantly increased the total root length. Similarly, 0.010 mmol/L SA significantly increased the root growth parameters of Z619 and Z39. In Z619, 0.1 mg/L PP333 significantly increased the total root length and total surface area and reduced the average root diameter. For all three cotton varieties, 0.050 mg/L GA3 increased peroxidase (POD) activity in the roots. In Z27 and Z39, 0.80 mg/L GA3 increased superoxide dismutase (SOD) activity in the roots. All SA concentrations increased SOD activity in roots of Z619 and Z27; 0.10 mg/L PP333 significantly increased SOD and POD activities in the roots of Z619 and significantly increased SOD activity in Z27. Principal component analysis indicated that 0.10 mmol/L SA was the optimal treatment for promoting the development of the roots of Z619 and 0.050 mmol/L SA was the optimal treatment for promoting the development of the roots of Z27 and Z39.

Endoplasmic reticulum stress promoted acinar cell necroptosis in acute pancreatitis through cathepsinB-mediated AP-1 activation.

Acinar cell death and inflammatory response are two important events which determine the severity of acute pancreatitis (AP). Endoplasmic reticulum (ER) stress and necroptosis are involved in this process, but the relationships between them remain unknown. Here, we analyzed the interaction between ER stress and necroptosis and the underlying mechanisms during AP. Experimental pancreatitis was induced in Balb/C mice by caerulein (Cae) and lipopolysaccharide (LPS) or L-arginine (L-Arg) in vivo, and pancreatic acinar cells were also used to follow cellular mechanisms during cholecystokinin (CCK) stimulation in vitro. AP severity was assessed by serum amylase, lipase levels and histological examination. Changes in ER stress, trypsinogen activation and necroptosis levels were analyzed by western blotting, enzyme-linked immunosorbent assay (ELISA), adenosine triphosphate (ATP) analysis or lactate dehydrogenase (LDH) assay. The protein kinase C (PKC)α -mitogen-activated protein kinase (MAPK) -cJun pathway and cathepsin B (CTSB) activation were evaluated by western blotting. Activating protein 1 (AP-1) binding activity was detected by electrophoretic mobility shift assay (EMSA). We found that ER stress is initiated before necroptosis in CCK-stimulated acinar cells in vitro. Inhibition of ER stress by 4-phenylbutyrate (4-PBA) can significantly alleviate AP severity both in two AP models in vivo. 4-PBA markedly inhibited ER stress and necroptosis of pancreatic acinar cells both in vitro and in vivo. Mechanistically, we found that 4-PBA significantly reduced CTSB maturation and PKCα-JNK-cJun pathway -mediated AP-1 activation during AP. Besides, CTSB inhibitor CA074Me markedly blocked PKCα-JNK-cJun pathway -mediated AP-1 activation and necroptosis in AP. However, pharmacologic inhibition of trypsin activity with benzamidine hydrochloride had no effect on PKCα-JNK-cJun pathway and necroptosis in CCK-stimulated pancreatic acinar cells. Furthermore, SR11302, the inhibitor of AP-1, significantly lowered tumor necrosis factor (TNF) α levels, and its subsequent receptor interacting protein kinases (RIP)3 and phosphorylated mixed lineagekinase domain-like (pMLKL) levels, ATP depletion and LDH release rate in CCK-stimulated pancreatic acinar cells. To sum up, all the results indicated that during AP, ER stress promoted pancreatic acinar cell necroptosis through CTSB maturation, thus induced AP-1 activation and TNFα secretion via PKCα-JNK-cJun pathway, not related with trypsin activity. These findings provided potential therapeutic target and treatment strategies for AP or other cell death-related diseases.

Dopamine D2 Receptor Signaling Attenuates Acinar Cell Necroptosis in Acute Pancreatitis through the Cathepsin B/TFAM/ROS Pathway.

Acute pancreatitis (AP) is an inflammatory disease that is associated with trypsinogen activation, mitochondrial dysfunction, cell death, and inflammation. Dopamine D2 receptor (DRD2) plays an essential role in alleviating AP, while it is unclear whether it is involved in regulating acinar cell necroptosis. Here, we found that DRD2 agonist quinpirole alleviated acinar cell necroptosis via inhibiting cathepsin B (CTSB). Moreover, CTSB inhibition by CA-074Me ameliorated AP severity by reducing necroptosis. Notably, knockdown of TFAM reversed the therapeutic effect of either quinpirole or CA-074Me. We identified a new mechanism that DRD2 signaling inhibited CTSB and promoted the expression of mitochondrial transcription factor A(TFAM), leading to reduction of ROS production in AP, which attenuated acinar cell necroptosis ultimately. Collectively, our findings provide new evidence that DRD2 agonist could be a new potential therapeutic strategy for AP treatment.

RELA-induced MiR-21 Exerts Oncogenic Effects on PDAC via Targeting of ARHGAP24.

Inflammation is one of the inducing factors of pancreatic ductal adenocarcinoma (PDAC), and microRNAs have been confirmed to be involved in the occurrence and development of PDAC. However, whether RELA, an inflammatory regulator, is involved in the regulation of PDAC by miRNA remains to be further studied. In the present study miR-21 was characterized and its upstream regulatory mechanism was investigated, as well as its functional effects and target genes in pancreatic ductal adenocarcinoma (PDAC). In situ hybridization analysis confirmed increased miR-21 expression levels in PDAC tissues. The results of the chromatin immunoprecipitation and dual-luciferase reporter assays demonstrated that transcription factor RELA modulated miR-21 transcription in the PDAC, PANC-1 and MIA PaCa-2 cell lines. Subsequently, a cell viability assay, EdU staining assay and flow cytometry analysis, demonstrated that miR-21 promoted cell proliferation and cell cycle progression, but inhibited cell apoptosis in vitro. Furthermore, a xenograft assay demonstrated that miR-21 accelerated tumor growth in vivo. Mechanistically, miR-21 directly regulated the expression of Rho GTPase activating protein 24 (ARHGAP24), which was indicated to be a tumor suppressor gene. Moreover, both miR-21 and ARHGAP24 were strongly associated with clinical features and may therefore serve as valuable biomarkers in PDAC prognosis.