AIM2 fosters lung adenocarcinoma immune escape by modulating PD-L1 expression in tumor-associated macrophages via JAK/STAT3.

This work was devised to discuss the effect of AIM2 on the immunosuppression of LUAD tumors, as well as its molecular mechanism. An allograft mouse model was built. Mouse macrophages were isolated and collected. The infiltration level of Mø and expression of M1 Mø, M2 Mø markers, and PD-L1 were assayed by IHC and flow cytometry. Expression levels of M1 Mø and M2 Mø marker genes and PD-L1 were detected by qPCR. The expression of proteins linked with JAK/STAT3 was tested by western blot. CD8+T cells and NK cells were activated in vitro and co-cultured with mouse macrophages, and their cytotoxicity was detected by LDH method. The proportion of CD206+PD-L1+ cells and the activation and proliferation of CD8+T cells were assayed by flow cytometry. Multicolor immunofluorescence was utilized to assay the co-localization of proteins. AIM2 demonstrated a high expression in LUAD, exhibiting a conspicuous positive correlation with the expression of the M2 Mø markers as well as PD-L1. Expression of M1 markers was upregulated after knockdown of AIM2, while M2 markers expression and PD-L1 were downregulated, and the colocalization of proteins linked with PD-L1 and M2 Mø was decreased. The infiltration and cytotoxicity of CD8+T cells and NK cells increased after silencing AIM2. After the knockdown of AIM2, which was enriched in the JAK/STAT3 pathway, the phosphorylation levels of JAK1, JAK2, and STAT3 were reduced, the immune infiltration level of CD8+T cells increased, and the co-localization level of PD-L1 and PD-1 dropped. The activity and proliferation level of CD8+T cells were increased with the reduced PD-1 expression. AIM2 fosters M2 Mø polarization and PD-L1 expression via the JAK/STAT3 pathway. Moreover, AIM2 promotes the immune escape of LUAD via the PD-1/PD-L1 axis. Our work may blaze a trail for the clinical treatment of LUAD.

3D human pose detection using nano sensor and multi-agent deep reinforcement learning.

Due to the complexity of three-dimensional (3D) human pose, it is difficult for ordinary sensors to capture subtle changes in pose, resulting in a decrease in the accuracy of 3D human pose detection. A novel 3D human motion pose detection method is designed by combining Nano sensors and multi-agent deep reinforcement learning technology. First, Nano sensors are placed in key parts of the human to collect human electromyogram (EMG) signals. Second, after de-noising the EMG signal by blind source separation technology, the time-domain and frequency-domain features of the surface EMG signal are extracted. Finally, in the multi-agent environment, the deep reinforcement learning network is introduced to build the multi-agent deep reinforcement learning pose detection model, and the 3D local pose of the human is output according to the features of the EMG signal. The fusion and pose calculation of the multi-sensor pose detection results are performed to obtain the 3D human pose detection results. The results show that the proposed method has high accuracy for detecting various human poses, and the accuracy, precision, recall and specificity of 3D human pose detection results are 0.97, 0.98, 0.95 and 0.98, respectively. Compared with other methods, the detection results in this paper are more accurate, and can be widely used in medicine, film, sports and other fields.

Diagnosing Coronavirus Disease 2019 (COVID-19): Efficient Harris Hawks-Inspired Fuzzy K-Nearest Neighbor Prediction Methods.

This study is devoted to proposing a useful intelligent prediction model to distinguish the severity of COVID-19, to provide a more fair and reasonable reference for assisting clinical diagnostic decision-making. Based on patients' necessary information, pre-existing diseases, symptoms, immune indexes, and complications, this article proposes a prediction model using the Harris hawks optimization (HHO) to optimize the Fuzzy K-nearest neighbor (FKNN), which is called HHO-FKNN. This model is utilized to distinguish the severity of COVID-19. In HHO-FKNN, the purpose of introducing HHO is to optimize the FKNN's optimal parameters and feature subsets simultaneously. Also, based on actual COVID-19 data, we conducted a comparative experiment between HHO-FKNN and several well-known machine learning algorithms, which result shows that not only the proposed HHO-FKNN can obtain better classification performance and higher stability on the four indexes but also screen out the key features that distinguish severe COVID-19 from mild COVID-19. Therefore, we can conclude that the proposed HHO-FKNN model is expected to become a useful tool for COVID-19 prediction.

Twelve out of 117 recovered COVID-19 patients retest positive in a single-center study of China.

BACKGROUND: It has been reported that a fraction of recovered coronavirus disease 2019(COVID-19) patients have retested positive for SARS-CoV-2. Clinical characteristics and risk factors for retesting positive have not been studied extensively. METHODS: In this retrospective, single-center cohort study, we included adult patients (≥ 18 years old) diagnosed as COVID-19 in Affiliated Yueqing Hospital, Wenzhou Medical University, Zhejiang, China. All the patients were discharged before March 31, 2020, and were re-tested for SARS-CoV-2 RNA by real-time reverse-transcriptase polymerase-chain-reaction (RT-PCR) after meeting the discharge criteria. We retrospectively analyzed this cohort of 117 discharged patients and analyzed the differences between retest positive and negative patients in terms of demographics, clinical characteristics, laboratory findings, chest computed tomography (CT) features and treatment procedures. FINDINGS: Compared with the negative group, the positive group had a higher proportion of patients with comorbidities (Odds Ratio(OR) =2·12, 95% Confidence Interval(CI) 0·48-9·46; p = 0·029), longer hospital stay (OR=1·21, 95% CI 1·07-1·36; p = 0·008), a higher proportion of patients with lymphocytopenia (p = 0·036), a higher proportion of antibiotics treatment (p = 0·008) and glucocorticoids treatment (p = 0·003). Multivariable regression showed increasing odds of positive SARS-CoV-2 retest after discharge associated with longer hospital stay (OR=1·22, 95% CI 1·08-1·38; p = 0·001), and lymphocytopenia (OR=7·74, 95% CI 1·70-35·21; p = 0·008) on admission. INTERPRETATION: Patients with COVID-19 who met discharge criteria could still test positive for SARS-CoV-2 RNA. Longer hospital stay and lymphopenia could be potential risk factors for positive SARS-CoV-2 retest in COVID-19 patients after hospital discharge. FUNDING: Natural Science Foundation of Zhejiang Province, Medical Scientific Research Fund of Zhejiang Province, Wenzhou science and technology project.

Melatonin attenuates the inflammatory response via inhibiting the C/EBP homologous protein-mediated pathway in taurocholate-induced acute pancreatitis.

Acute pancreatitis (AP) is a serious disease characterized by the activation of trypsin, autodigestion, edemas, hemorrhages and necrosis. However, the mechanisms of regulating the apoptosis and inflammation of acinar cells in AP remain unclear. The endoplasmic reticulum (ER) stress­related molecule, C/EBP homologous protein (CHOP), has pro­-apoptotic and pro­inflammatory properties, in addition to regulating ER stress responses. In the present study, a lentivirus­mediated RNA interference (RNAi) approach was used to specifically knockdown the expression of CHOP in the pancreatic tissue of Sprague­Dawley rats to investigate the potential role of CHOP during AP, which was induced by the retrograde injection of 5% taurocholate into the biliopancreatic duct of rats. Pre­treatment with melatonin was further used to identify the potential anti­inflammatory mechanisms in AP. Pancreatic tissues were procured for western blot analysis, histological examination, reverse transcription­quantitative polymerase chain reaction and immunohistochemical staining. ER stress was rapidly activated in the early stage and increased over time in the rat AP model. However, the silencing of CHOP expression markedly inhibited apoptosis and ER stress, reducing the activation of nuclear factor­κB and inflammation injury in AP. Melatonin also exhibited anti­inflammatory and apoptotic effects, and significantly decreased the expression of CHOP. Thus, it can be concluded that the CHOP­mediated pathway serves an important role in the development of AP, and that melatonin can reduce pancreatic damage via the inhibition of CHOP expression.

A retrospective and prospective assessment of the zero-markup drug reform in China from the perspective of policy diffusion.

The zero-markup drug policy is an important component of the new round of Chinese health care reform that began in 2009 to promote the separation between medical and pharmaceutical services, reduce patients' medical burden, and improve the medical supply security system. Over the past 8 years, the zero-markup drug reform policy has been carried out in 4 pilot rounds (a policy diffusion model with Chinese characteristics) and has been promoted throughout the mainland China. At this critical point, it is necessary to review this policy systematically. Therefore, based on the literature, government documents, and interview records, this study analyzed the characteristics, progress, achievements, challenges, and recommendations of zero-markup drug reform by using the policy diffusion theory. The study found that zero-markup drug reform has completed its initial diffusion by use of the "policy experiment" method and has reduced drug prices and patients' burden to a certain extent. However, in the next phase of policy diffusion, the reform still requires adjustment and innovative measures to respond to future challenges. Generally speaking, as China's unique health care reform practice, the experience of zero-markup drug reform could be used as a reference for other countries to control drug prices, separate medical and pharmaceutical services, and establish a modern system of hospital operation.

Melatonin Attenuates Endoplasmic Reticulum Stress in Acute Pancreatitis.

OBJECTIVES: The objective of this study is to explore the effect of melatonin on endoplasmic reticulum stress in acute pancreatitis (AP) and the molecular mechanism. METHODS: Acute pancreatitis was induced in vivo in Sprague-Dawley rats by the retrograde injection of 5% taurocholate into the biliopancreatic duct and in vitro by treating AR42J cells with cerulein (10 nmol/L) plus lipopolysaccharide (LPS) (10 mg/L). The rats and cells were treated with melatonin (50 mg/kg in rats and 0.5, 1, and 2 mmol/L in AR42J cells) 30 minutes before AP was induced. After 9 hours, the cells and rat pancreas tissue were collected for Western blot, reverse transcription polymerase chain reaction, histological examination, immunohistochemistry, and immunofluorescence analysis. RESULTS: Inositol-requiring 1α (IRE1α)-mediated Jun N-terminal kinase (JNK)/nuclear factor-kappa B (NF-κB) pathway were activated early in AR42J cells and rat AP models. Melatonin significantly inhibited the expression of proinflammatory cytokines. Western blot and immunohistochemical results all indicated that melatonin regulated apoptosis-related protein expression. In addition, melatonin treatment resulted in significantly reduced pancreatic tissue injury, as revealed by histological changes and pathological scores. Furthermore, melatonin treatment significantly reduced the activation of IRE1α-mediated JNK/NF-κB pathway-related proteins. CONCLUSIONS: These findings suggest that melatonin protects AR42J cells and Sprague-Dawley rats against AP-associated injury, probably through downregulation of IRE1α-mediated JNK/NF-κB pathways.

Melatonin induces anti-inflammatory effects via endoplasmic reticulum stress in RAW264.7 macrophages.

Melatonin, which is predominantly secreted by the pineal gland and is released into the blood, appears to have anti­inflammatory properties. Several studies have shown that melatonin can relieve lipopolysaccharide-induced inflammatory responses of RAW264.7 cells. However, the mechanisms underlying this anti­inflammatory effect remain to be fully elucidated, particularly the association between melatonin and endoplasmic reticulum (ER) stress (ERS). Therefore, the present study examined the anti­inflammatory activity of melatonin in RAW264.7 cells and analyzed its molecular mechanisms in ERS. The RAW264.7 cells were stimulated by lipopolysaccharide and treated with melatonin. A Cell Counting Kit­8 assay was used to assess the toxicity of melatonin. The degree of inflammation was evaluated using ELISA. The expression levels of ERS­associated protein molecules were examined using reverse transcription­quantitative polymerase chain reaction and western blot analyses. The results revealed that melatonin had no toxic effect on the RAW264.7 cells at the range of concentrations used in the experiment. Lipo-polysaccharide stimulated the cells to produce inflammatory molecules; in the early stage, proteins associated with ERS increased, and then apoptosis occurred. The cells treated with melatonin exhibited attenuated inflammation, decreased expression of ERS­associated proteins and inhibition of apoptosis. Taken together, the results of the present study showed that melatonin may attenuate the inflammatory response by inhibiting the activation of ERS in RAW264.7 macrophages.

Impact of non-alcoholic fatty liver disease and smoking on colorectal polyps.

OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) and smoking have similar mechanisms of promoting colorectal polyps. The potential link between NAFLD and smoking in men and colorectal polyps has not been adequately evaluated. The aim is to investigate this association. METHODS: A retrospective cross-sectional study was conducted on 2409 individuals undergoing a health check. Univariate and multivariate logistic regression were performed for analyzing the association between risk factors and colorectal polyps. Individuals were divided into four groups: Q1: NAFLD (-)/smoking (-); Q2: NAFLD (+)/smoking (-); Q3: NAFLD (-)/smoking (+); Q4: NAFLD (+)/smoking (+). Logistic analyses were used to explore associations for the whole study population and stratified groups. RESULTS: The prevalence of colorectal polyps was 38.8% in males, and that of colorectal polyps in smokers and individuals with NAFLD were 47.0% (428/911) and 42.9% (267/622), respectively. With Q1 as reference, subjects with NAFLD (+) and smoking habits (+) had the highest ORs for colorectal polyps (OR = 2.64, 95% CI: 1.91 - 3.64, P < 0.001), adenomatous polyps (OR = 2.06, 95% CI: 1.38 - 3.05, P < 0.05), non-adenomatous polyps (OR = 1.97, 95% CI: 1.39 - 2.80, P < 0.05), ≥ 3 polyps (OR = 2.05, 95% CI: 1.31 - 3.22, P < 0.05) and proximal polyps (OR = 1.58, 95% CI: 1.02 - 2.45, P < 0.05) after adjusting for confounding variables. CONCLUSIONS: Men with NAFLD and smoking habits have an increasing risk of colorectal polyps.

Sex-influenced association of non-alcoholic fatty liver disease with colorectal adenomatous and hyperplastic polyps.

AIM: To investigate the relationship between non-alcoholic fatty liver disease (NAFLD) and colorectal adenomatous and hyperplastic polyps. METHODS: A retrospective cross-sectional study was conducted on 3686 individuals undergoing health checkups (2430 males and 1256 females). All subjects underwent laboratory testing, abdominal ultrasonography, colonoscopy, and an interview to ascertain the baseline characteristics and general state of health. Multinomial logistic regression analysis was performed to examine the association between NAFLD and the prevalence of colorectal adenomatous and hyperplastic polyps. Furthermore, the relationship was analyzed in different sex groups. Subgroup analysis was performed based on number, size, and location of colorectal polyps. RESULTS: The prevalence of colorectal polyps was 38.8% in males (16.2% for adenomatous polyps and 9.8% for hyperplastic polyps) and 19.3% in females (8.4% for adenomatous polyps and 3.9% for hyperplastic polyps). When adjusting for confounding variables, NAFLD was significantly associated with the prevalence of adenomatous polyps (OR = 1.28, 95%CI: 1.05-1.51, P < 0.05) and hyperplastic polyps (OR = 1.35, 95%CI: 1.01-1.82, P < 0.05). However, upon analyzing adenomatous and hyperplastic polyps in different sex groups, the significant association remained in males (OR = 1.53, 95%CI: 1.18-2.00, P < 0.05; OR = 1.42, 95%CI: 1.04-1.95, P < 0.05) but not in females (OR = 0.44, 95%CI: 0.18-1.04, P > 0.05; OR = 1.18, 95%CI: 0.50-2.78, P > 0.05). CONCLUSION: NAFLD is specifically associated with an increased risk of colorectal adenomatous and hyperplastic polyps in men. However, NAFLD may not be a significant factor in the prevalence of colorectal polyps in women.

Melatonin Induces Anti-Inflammatory Effects to Play a Protective Role via Endoplasmic Reticulum Stress in Acute Pancreatitis.

BACKGROUND/AIMS: Melatonin, which is mainly secreted by the pineal gland and released into blood, has anti-inflammatory properties in acute pancreatitis. Many studies show that melatonin can relieve inflammation in taurocholate-induced acute pancreatitis. However, the mechanisms of its anti-inflammatory effects are still undefined, especially the relationship between melatonin and endoplasmic reticulum stress. We explored the anti-inflammatory activity of melatonin in AR42J and rat models. METHODS: The CCK-8 assay was used to assess effects of melatonin on AR42J cell viability. Inflammatory degree and the expressions of endoplasmic reticulum stress related molecules were examined by quantitative RT-PCR and western blotting. The degree of inflammation in the tissue was also accessed by pathological grading. Finally, we used the western blotting method to verify apoptosis and autophagy. RESULTS: Endoplasmic reticulum stress was obviously activated in early stage inflammation in AR42J and rat models. Melatonin could induce anti-inflammatory effects via endoplasmic reticulum stress. Melatonin significantly inhibited inflammatory cytokines and the expression of ERS-related molecules. Finally, it played a protective role by promoting apoptosis and autophagy of the cells, which were damaged in the process of inflammatory reaction. CONCLUSION: Melatonin induces anti-inflammatory effects via endoplasmic reticulum stress in acute pancreatitis to play a protective role.