Factors associated with non-lifting of colorectal mucosal lesions.

BACKGROUND: Endoscopic submucosal dissection (ESD) is an effective treatment for colorectal tumors. However, lesions that cannot be lifted after submucosal injection are not indication for ESD. This is because the procedure is difficult, and the lesions are often considered as tumor invasion or submucosal fibrosis. The aims of this study are to evaluate the efficacy and safety of ESD for non-lifting lesions and to analyze the causes of non-lifting phenomenon. METHODS: This retrospective study included 29 patients with non-lifting colon lesions resected by ESD from February 2018 to September 2021. Cases were observed for demographics, endoscopic findings, treatment outcomes, adverse events and endoscopic follow-up. We studied the pathological features of lesions to explore the reasons for non-lifting. RESULTS: Among 29 cases of non-lifting lesions, 20 lesions (69.0%) were 30 mm in diameter or larger. Most of lesions (96.6%) were non-lifting in center, and only one lesions (3.4%) had non-lifting of one side. The en bloc and curative resection rates of ESD were 100 and 86.2%, respectively. There was one (3.4%) delayed bleeding, no perforations and other complications. No tumor recurrence occurred during the follow-up period. For pathological features, 16 (55.2%) non-lifting lesions had submucosal fibrosis and only 4 cases (13.8%) had deep submucosal invasion. There were 9 cases (31.0%) of non-lifting lesions due to musculo-fibrous of muscularis propria anomaly (MMPA). CONCLUSION: MMPA is another reason for non-lifting signs besides invasive carcinomas and submucosal fibrosis. ESD should be considered in patients with large non-lifting adenoma instead of surgery.

Effectiveness and safety of regimens containing linezolid for treatment of Mycobacterium abscessus pulmonary Disease.

OBJECTIVE: To evaluate the effectiveness and safety of linezolid-containing regimens for treatment of M. abscessus pulmonary disease. METHODS: The records of 336 patients with M. abscessus pulmonary disease who were admitted to Shanghai Pulmonary Hospital from January 2018 to December 2020 were retrospectively analyzed. A total of 164 patients received a linezolid-containing regimen and 172 controls did not. The effectiveness, safety, antibiotic susceptibility profiles, outcomes, culture conversion, cavity closure, and adverse reactions were compared in these two groups. RESULTS: The two groups had similar treatment success (56.1% vs. 48.8%; P > 0.05), but treatment duration was shorter in the linezolid group (16.0 months [inter-quartile ranges, IQR: 15.0-17.0] vs. 18.0 months [IQR: 16.0-18.0]; P < 0.01). The rates of sputum culture conversion were similar (53.7% vs. 46.5%, P > 0.05), but time to conversion was shorter in the linezolid group (3.5 months [IQR: 2.5-4.4] vs. 5.5 months [IQR: 4.0-6.8]; P < 0.01). The linezolid group had a higher rate of cavity closure (55.2% vs. 28.6%, P < 0.05) and a shorter time to cavity closure (3.5 months [IQR: 2.5-4.4] vs. 5.5 months [IQR: 4.0-6.8]; P < 0.01). Anemia and peripheral neuropathy were more common in the linezolid group (17.7% vs. 1.7%, P < 0.01; 12.8% vs. 0.6%, P < 0.01). CONCLUSIONS: The linezolid and control groups had similar treatment success rates. The linezolid group had a shorter treatment duration, shorter time to sputum culture conversion, and higher rate and shorter time to lung cavity closure. More patients receiving linezolid developed anemia and peripheral neuropathy.

RETRACTED: LncRNA NEAT1 participates in inflammatory response in macrophages infected by mycobacterium tuberculosis through targeted regulation of miR-377-3p.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor-in-Chief. After a thorough investigation, the Editor has concluded that the acceptance of this article was partly based upon the positive advice of one illegitimate reviewer report. The report was submitted from an email account which was provided to the journal as a suggested reviewer during the submission of the article. Although purportedly a real reviewer account, the Editor has concluded that this was not of an appropriate, independent reviewer. This manipulation of the peer-review process represents a clear violation of the fundamentals of peer review, our publishing policies, and publishing ethics standards. Apologies are offered to the reviewer whose identity was assumed and to the readers of the journal that this deception was not detected during the submission process.

Activation of Nrf2 signaling by oltipraz inhibits death of human macrophages with mycobacterium tuberculosis infection.

Mycobacterium tuberculosis (MTB) infection can induce cytotoxicity to the host macrophages, promoting bacterial spread. We here tested the potential effect of oltipraz, a synthetic dithiolethione, in MTB-infected human macrophages. We show that oltipraz significantly inhibited MTB-induced death and apoptosis in human macrophages. MTB-induced reactive oxygen species production, mitochondrial depolarization and programmed necrosis were attenuated by oltipraz in macrophages. Oltipraz activated Nrf2 signaling, causing Keap1-Nrf2 disassociation, Nrf2 protein stabilization and nuclear translocation, simultaneously promoting expression of Nrf2-dependent genes (HO1, NQO1 and GST) in human macrophages. Nrf2 shRNA or CRISPR/Cas9-induced Nrf2 knockout completely reversed oltipraz-induced macrophage protection against MTB infection. Furthermore, CRISPR/Cas9-mediated Keap1 knockout induced Nrf2 cascade activation and protected human macrophages from MTB. Importantly, oltipraz was unable to offer further cytoprotection against MTB in Keap1 knockout macrophages. Collectively we conclude that oltipraz activates Nrf2 signaling cascade to protect human macrophages from MTB-induced oxidative injury and cell death.

LncRNA MEG3 control Mycobacterium Tuberculosis infection via controlled MiR-145-5p expression and modulation of macrophages proliferation.

BACKGROUND: Mycobacterium tuberculosis (Mtb) is an intractable pathogen for humans to overcome. While as an important part of innate immunity, macrophages play an important role in resisting foreign pathogenic microorganisms, and it has been proved that there is a close relationship between macrophages and Mtb. In recent years, with the in-depth study of LncRNA, there have been crucial breakthroughs in the diagnosis and treatment of a number of diseases, and understanding the impact of LncRNA on Mtb may also be conducive to providing new therapeutic targets for tuberculosis prevention and treatment in the future. Therefore, this study explore the role of MEG3 in the proliferation and apoptosis of Mtb-infected macrophages. METHODS: Between September 2017 and September 2019, 84 consecutive pulmonary Mtd patients admitted to our hospital were selected as the observation group (OG), and concurrently, 88 healthy controls were selected as the control group (CG). MEG3 and miR-145-5p in peripheral blood of the two groups were detected, and their diagnostic value in pulmonary tuberculosis (PTB) was analyzed by receiver operating characteristic (ROC) curves. In addition, Bacillus Calmette-Guérin (BCG) and mouse Raw264.7 macrophage strains were purchased to establish the Mtb-infected macrophage model. Colony forming unit (CFU) and flow cytometry were employed to determine the effects of MEG3 and miR-145-5p on macrophages, and the correlation between the two was performed by dual-luciferase reporter (DLR) assay. RESULTS: MEG3 was highly expressed in PTB, while miR-145-5p was lowly expressed (P < 0.050). ROC analysis demonstrated that both MEG3 and miR-145-5p enjoyed favorable predictive value for the occurrence of PTB (P < 0.001). In the Mtb-infected macrophage model, MEG3 decreased (P < 0.050) while miR-145-5p increased with the time of infection (P < 0.050). Inhibited MEG3 and overexpressed miR-145-5p resulted in increased CFU and decreased apoptosis ability of macrophages (P < 0.050). The DLR assay identified a targeting relationship between miR-145-5p and MEG3 (P < 0.050). The increase of MEG3 could inhibit the expression of in miR-145-5p (P < 0.050). CONCLUSION: MEG3 affects the biological activity of Mtb-infected macrophages by targeting miR-145-5p, which may be the key to the diagnosis and treatment of PTB and even all kinds of tuberculosis in the future.

Z-Score Reference Ranges for the Offset of the Tricuspid Septal Leaflet in Normal Fetuses.

OBJECTIVES: To determine Z-score equations and reference ranges for mitral valve-tricuspid valve distance (MTD) and the MTD index in the fetal heart. METHODS: A prospective cross-sectional study was performed in 899 normal singleton fetuses from 14 to 40 weeks' gestation. The MTD and interventricular septum length (IVSL) were measured offline after electronic cardiac spatiotemporal image correlation volume acquisition. The MTD index was determined as the ratio of MTD to IVSL. Z-score reference ranges of these measurements were determined against gestational age (GA) and estimated fetal weight (EFW), using regression analysis of the mean and standard deviation (SD). RESULTS: Strong positive correlations were found between the MTD and the independent variables. A simple linear regression model was the best description of the mean and SD of MTD based on GA, while a cubic regression best fitted the mean MTD against EFW. In contrast, the MTD index decreased progressively with the independent variables. Fractional polynomials best fitted the MTD index in terms of GA and EFW. CONCLUSION: Normal reference values and Z-scores of fetal MTD and MTD index were provided against GA and EFW, which may be useful tools for quantitative assessment of some cardiac and extracardiac diseases.

Extract of Paecilomyces hepiali mycelia induces lipolysis through PKA-mediated phosphorylation of hormone-sensitive lipase and ERK-mediated downregulation of perilipin in 3T3-L1 adipocytes.

BACKGROUND: Cordyceps sinensis has been used for centuries in China as one of the most valued herbal medicine and tonic food. Paecilomyces hepiali, a fungal strain isolated from natural C. sinensis, has been used widely as a substitute of C. sinensis in medicine and health food. P. hepiali has been reported to have various pharmaceutical benefits, including triglyceride-lowing activity. However, its effects on triglyceride metabolism in adipocytes remain unknown. The purpose of the present study was to evaluate the effect of P. hepiali mycelia on adipocyte lipolysis and to clarify the underlying mechanisms. METHODS: The fully differentiated 3T3-L1 adipocytes were treated with methanol extract of Paecilomyces hepiali mycelia (PHME). Contents of glycerol released into the culture medium and intracellular triglyceride were measured as indices of lipolysis using glycerol assay kit and Oil red O staining, respectively. Then, effects of PHME on the main lipases or kinases involved in lipolysis regulation were investigated. Protein expression of adipose triglyceride lipase (ATGL) and perilipin, as well as phosphorylation of hormone-sensitive lipase (HSL), AMP-activated protein kinase (AMPK), and mitogen-activated protein kinases (MAPKs) were determined by western blotting. Moreover, nucleosides, important constituents of PHME, were analyzed using high performance liquid chromatography (HPLC). RESULTS: Treatment with PHME led to a significant increase in glycerol release thereby reduced intracellular triglyceride accumulation in fully differentiated adipocytes. PHME upregulated protein kinase (PK) A-mediated phosphorylation of HSL at serine residues of 563 and 660. Meanwhile, PHME treatment also upregulated phosphorylation of extracellular signal-regulated kinase (ERK), and downregulated the protein level of perilipin. Pretreatment with the PKA inhibitor, H89, blunted the PHME-induced lipolysis and the phosphorylation of HSL (Ser 563 and 660). Moreover, pretreatment with ERK inhibitor, PD98059, weakened the PHME-caused glycerol release and downregulation of perilipin expression. HPLC analysis indicated there were adenosine, cordycepin, uridine and vernine in PHME. CONCLUSIONS: Our results showed that PHME significantly induced lipolysis in 3T3-L1 adipocytes, which is mainly mediated by activation of HSL through PKA pathway and by downregulation of perilipin through activation of ERK pathway.

Investigating genetic characteristics of hepatitis B virus-associated and -non-associated hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is a primary liver malignancy that mainly occurs in patients with chronic liver disease and cirrhosis. Risk factors for HCC include hepatitis B virus (HBV) infection. However, the specific role of HBV infection in HCC development is not yet completely understood. In order to reveal the effects of HBV on HCC, we compare the genes of HCC patients infected with HBV with those who are not infected. METHODS: We encoded the genes of these two types of HCC in databases using enrichment scores of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway terms. A random forest algorithm was employed in order to distinguish these two types in the classifier, and a series of feature selection approaches was used in order to select their optimal features. Novel HBV-associated and -non-associated HCC genes were predicted, respectively, based on their optimal features in the classifier. A shortest-path algorithm was also employed in order to find all of the shortest-paths genes connecting the known related genes. RESULTS: A total of 54 different features between HBV-associated and -non-associated HCC genes were identified. In total, 1236 and 881 novel related genes were predicted for HBV-associated and -non-associated HCC, respectively. By integrating the predicted genes and shortest path genes in their gene interaction network, we identified 679 common genes involved in the two types of HCC. CONCLUSION: We identified the significantly different genetic features between two types of HCC. We also predicted related genes for the two types based on their specific features. Finally, we determined the common genes and features that were involved in both of these two types of HCC.

The impact of cranioplasty on cerebral blood perfusion in patients treated with decompressive craniectomy for severe traumatic brain injury.

BACKGROUND: A large cranial defect following decompressive craniectomy (DC) is a common sequela in patients with severe traumatic brain injury (TBI). Such a defect can cause severe disturbance of cerebral blood flow (CBF) regulation. This study investigated the impact of cranioplasty on CBF in these patients. METHODS: Patients who underwent DC and secondary cranioplasty were prospectively studied for a severe TBI. CT perfusion was used to measure CBF before and after cranioplasty. The basal ganglia, parietal lobe and occipital lobe on the decompressed side were chosen as zones of interest for CBF evaluation. RESULTS: Nine patients representing nine cranioplasty procedures were included in the study. Before cranioplasty, CBF on the decompressed side was lower than that on the contralateral side. During the early stage (10 days) after cranioplasty, CBF on the decompressed side was increased and this increase was significant in the parietal and occipital lobe. CBF was also increased on the contralateral side. In addition, the difference in CBF between the contralateral side and the decompressed side was reduced after cranioplasty. Further, the CT perfusion showed that the CBFs decreased again 3 months post-cranioplasty among four cases, but was still higher than those before cranioplasty. CONCLUSIONS: This study indicates that cranioplasty may increase CBF and benefit the recovery in patients with DC for TBI.

The novel mTORC1/2 dual inhibitor INK-128 suppresses survival and proliferation of primary and transformed human pancreatic cancer cells.

Pancreatic cancer has one of worst prognosis among all human malignancies around the world, the development of novel and more efficient anti-cancer agents against this disease is urgent. In the current study, we tested the potential effect of INK-128, a novel mammalian target of rapamycin (mTOR) complex 1 and 2 (mTORC1/2) dual inhibitor, against pancreatic cancer cells in vitro. Our results demonstrated that INK-128 concentration- and time-dependently inhibited the survival and growth of pancreatic cancer cells (both primary cells and transformed cells). INK-128 induced pancreatic cancer cell apoptosis and necrosis simultaneously. Further, INK-128 dramatically inhibited phosphorylation of 4E-binding protein 1 (4E-BP1), ribosomal S6 kinase 1 (S6K1) and Akt at Ser 473 in pancreatic cancer cells. Meanwhile, it downregulated cyclin D1 expression and caused cell cycle arrest. Finally, we found that a low concentration of INK-128 significantly increased the sensitivity of pancreatic cancer cells to gemcitabine. Together, our in vitro results suggest that INK-128 might be further investigated as a novel anti-cancer agent or chemo-adjuvant for pancreatic cancer treatment.

[Sonographic features and diagnostic analysis of breast granulomatous diseases: a report of 32 cases].

OBJECTIVE: To evaluate the sonographic features of different pathological types of breast granulomatous diseases and analyze the feasibility of ultrasonic diagnosis. METHODS: A total of 32 patients with different pathological types of breast granulomatous diseases were recruited. Their clinical and sonographic findings were retrospectively reviewed. There were granulomatous mastitis (n = 12), breast xanthogranuloma (n = 5), lipogranuloma (n = 2), foreign body granuloma (n = 1) and nonspecific granulation hyperplasia (n = 12). RESULTS: Based on major sonographic appearances, they were divided into 4 patterns of tubular, mass, diffuse and cystic mass. In 12 patients with granulomatous mastitis and 12 patients with nonspecific granulation hyperplasia, the major sonographic appearance was of tubular pattern (n = 6, 5), followed by mass pattern (n = 4, 5) and diffuse pattern (n = 2, 2). Five patients with breast xanthogranuloma and 1 patient with foreign body granuloma all showed mass pattern. In 2 patients with lipogranuloma, one was of mass pattern and another cystic pattern. In patients with granulomatous mastitis and patients with nonspecific granulation hyperplasia, it showed a high diagnostic reliability of ultrasound. The ratio of inflammatory lesion as the first sonographic diagnosis was 10/12 and 8/12 respectively and ultrasonic BI-RADS 4b or above both only 1/12. However, the ratio of sonographic imaging in patients with xanthogranuloma and Lipogranuloma mimic breast cancer, in which ultrasonic score as breast imaging-reporting and data system (BI-RADS) 4b or above was 4/5 and 1/1 respectively. CONCLUSIONS: Ultrasound is valuable in evaluating the lesions in patients with granulomatous mastitis and nonspecific granulation hyperplasia. However a definite diagnosis is still dependent on histopathology.

Clinical Features of Nontuberculous Mycobacterial Pulmonary Disease in the Yangtze River Delta of China: A Single-Center, Retrospective, Observational Study.

With increased focus on nontuberculous mycobacterial pulmonary disease (NTM-PD), and the improvement in detection methods, the global incidence continues to increase every year, but the diagnosis and treatment are difficult with a high misdiagnosis rate and poor curative effect. This study aimed to analyze the clinical indicators of different pathogenic NTM in the Yangtze River Delta. The study retrospectively analyzed the medical records of patients with NTM-PD, who resided in the Yangtze River Delta and were diagnosed using sputum or bronchial lavage fluid and hospitalized in Shanghai Pulmonary Hospital from March 2017 to February 2019. The clinical data of confirmed patients were collected. Among the 513 cases of NTM-PD, 482 cases were infected by four common bacteria: Mycobacterium intracellulare (224, 46.5%), M. abscessus (138, 28.6%), M. kansasii (84, 17.4%), and M. avium (36, 7.5%). The analysis found that different NTM strains have their corresponding positive and negative correlation factors (p < 0.05). M. intracellulare, M. abscessus, M. kansasii, and M. avium were the main pathogenic bacteria isolated from patients with NTM-PD in the Yangtze River Delta were. Different strains resulted in different clinical features, assisting in the early diagnosis and treatment of NTM-PD.

Pelvic floor dysfunction and electrophysiology in postpartum women at 6-8 weeks.

Objective: To investigate the incidence of pelvic floor dysfunction (PFD) and electrophysiological indicators in postpartum women at 6-8 weeks and explore the influence of demographic characteristics and obstetric factors. Methods: A survey questionnaire collected information about the conditions of women during their pregnancy and puerperal period and their demographic characteristics; pelvic organ prolapse quantitation (POP-Q) and pelvic floor muscle electrophysiology (EP) examination were conducted in postpartum women at 6-8 weeks. Results: Vaginal delivery was a risk factor for anterior pelvic organ prolapse (POP) (OR 7.850, 95% confidence interval (CI) 5.804-10.617), posterior POP (OR 5.990, 95% CI 3.953-9.077), anterior and posterior stage II POP (OR 6.636, 95% CI 3.662-15.919), and postpartum urinary incontinence (UI) (OR 6.046, 95% CI 3.894-9.387); parity was a risk factor for anterior POP (OR 1.397,95% CI 0.889-2.198) and anterior and posterior stage II POP (OR 4.162, 95% CI 2.125-8.152); age was a risk factor for anterior POP (OR 1.056, 95% CI 1.007-1.108) and postpartum UI (OR 1.066, 95% CI 1.014-1.120); body mass index (BMI) was a risk factor for postpartum UI (OR 1.117, 95% CI 1.060-1.177); fetal birth weight was a risk factor for posterior POP (OR 1.465, 95% CI 1.041-2.062); and the frequency of pregnancy loss was a risk factor for apical POP (OR 1.853, 95% CI 1.060-3.237). Conclusion: Pelvic floor muscle EP is a sensitive index of early pelvic floor injury. The changes in muscle strength and fatigue degree coexist in different types of postpartum PFD, and each has its own characteristics.

Efficacy and safety of an innovative short-course regimen containing clofazimine for treatment of drug-susceptible tuberculosis: a clinical trial.

In preclinical studies, a new antituberculosis drug regimen markedly reduced the time required to achieve relapse-free cure. This study aimed to preliminarily evaluate the efficacy and safety of this four-month regimen, consisting of clofazimine, prothionamide, pyrazinamide and ethambutol, with a standard six-month regimen in patients with drug-susceptible tuberculosis. An open-label pilot randomized clinical trial was conducted among the patients with newly diagnosed bacteriologically-confirmed pulmonary tuberculosis. The primary efficacy end-point was sputum culture negative conversion. Totally, 93 patients were included in the modified intention-to-treat population. The rates of sputum culture conversion were 65.2% (30/46) and 87.2% (41/47) for short-course and standard regimen group, respectively. There was no difference on two-month culture conversion rates, time to culture conversion, nor early bactericidal activity (P > 0.05). However, patients on short-course regimen were observed with lower rates of radiological improvement or recovery and sustained treatment success, which was mainly attributed to higher percent of patients permanently changed assigned regimen (32.1% vs. 12.3%, P = 0.012). The main cause for it was drug-induced hepatitis (16/17). Although lowering the dose of prothionamide was approved, the alternative option of changing assigned regimen was chosen in this study. While in per-protocol population, sputum culture conversion rates were 87.0% (20/23) and 94.4% (34/36) for the respective groups. Overall, the short-course regimen appeared to have inferior efficacy and higher incidence of hepatitis but desired efficacy in per-protocol population. It provides the first proof-of-concept in humans of the capacity of the short-course approach to identify drug regimens that can shorten the treatment time for tuberculosis.

Liver transplantation in Crigler-Najjar syndrome type I disease.

BACKGROUND: Crigler-Najjar syndrome type I (CNS I) is a very rare autosomal recessive inherited disease that liver transplantation can properly deal with. METHODS: We present one case of an 18-month-old child with CNS I diagnosed by clinical findings and genetic detecting. LTx was performed 5 days after kernicterus broke out and neurological symptoms were successfully reversed. RESULT: Magnetic resonance imaging and magnetic resonance spectroscopy showed encouraging results that brain pathology had a trend to return to normal in 1-year follow-up, combined with electroencephalogram and motor development estimate studies. CONCLUSIONS: Liver transplantation can cure CNS I with reversible neurological symptoms to some extent in time. Magnetic resonance spectroscopy may be a future option of predicting brain conditions and selecting suitable patients with CNS I for transplantation.

Factors Associated with Treatment Outcome in Patients with Nontuberculous Mycobacterial Pulmonary Disease: A Large Population-Based Retrospective Cohort Study in Shanghai.

Infectious diseases caused by nontuberculous mycobacteria (NTM) are increasingly common. This retrospective cohort study examined factors associated with outcomes in patients from Shanghai who had NTM pulmonary disease (NTMPD) from January 2014 to December 2018. The causative bacterial species, drug susceptibility test results, treatment outcomes, sputum culture conversion rate, and risk factors associated with treatment failure were determined. The most common species were Mycobacterium avium complex (MAC) (50%), M. abscessus (28%), and M. kansasii (15%). Over five years, the proportions of M. kansasii and M. abscessus increased, and that of MAC decreased. The treatment success rate was significantly greater for patients infected with M. kansasii (89.9%) than MAC (65.0%, p < 0.001) and M. abscessus (36.1%, p < 0.001). Multivariate analysis indicated the risk factors for treatment failure were pathogenic NTM species (M. abscessus: aOR = 9.355, p < 0.001; MAC: aOR = 2.970, p < 0.001), elevated ESR (>60 mm/h: aOR = 2.658, p < 0.001), receipt of retreatment (aOR = 2.074, p < 0.001), and being middle-aged or elderly (>60 years-old: aOR = 1.739, p = 0.021; 45-60 years-old: aOR = 1.661, p = 0.034). The main bacterial species responsible for NTMPD were MAC, M. abscessus, and M. kansasii. Patients who were infected by M. abscessus or MAC, with elevated ESR, received retreatment, and were middle-aged or elderly had an increased risk of treatment failure.

Integrin α4ß1/VCAM-1 Interaction Evokes Dynamic Cell Aggregation Between Immune Cells and Human Lung Microvascular Endothelial Cells at Infectious Hemolysis.

Bacterial and viral infection is a common cause of pneumonia, respiratory failure, and even acute respiratory distress syndrome. Increasing evidence indicates that red blood cells (RBCs) may contribute to immune response and inflammation. However, the precise molecular mechanisms that link RBC and hemolysis to the development and progression of inflammatory pathologies are not entirely understood. In this study, we used bacterial endotoxin, lipopolysaccharide (LPS), to mimic an infectious hemolysis and found that RBCs dynamically regulated cell aggregation between immune cells and human lung microvascular endothelial cells (HLMVEC). When RBCs were treated with LPS, integrin α4ß1 was increased and was accompanied by cytokines and chemokines release (TNF-α, IL-1ß, IL-6, IL-8, IFN-γ, CXCL12, CCL5, CCL7 and CCL4). Upon α4ß1 elevation, RBCs not only facilitated mature monocyte derived dendritic cell (mo-DCs) adhesion but also promoted HLMVEC aggregation. Furthermore, co-culture of the supernatant of LPS pre-treated RBCs with mo-DCs could promote naïve CD4 T cell proliferation. Notably, the filtered culture from LPS-lysed RBCs further promoted mo-DCs migration in a concentration dependent manner. From a therapeutic perspective, cyclic peptide inhibitor of integrin α4ß1 combined with methylprednisolone (α4ß1/Methrol) remarkably blocked RBCs aggregation to mo-DCs, HLMVEC, or mo-DCs and HLMVEC mixture. Moreover, α4ß1/Methrol dramatically reduced mo-DCs migration up-regulated glucocorticoid-induced leucine zipper in mo-DCs, and ultimately reversed immune cell dysfunction induced by hemolysis. Taken together, these results indicate that integrin α4ß1 on RBCs could mediate cell-cell interaction for adaptive immunity through influencing cell adhesion, migration, and T cell proliferation.

Alisol A 24-acetate stimulates lipolysis in 3 T3-L1 adipocytes.

BACKGROUND: Alisol A 24-acetate (AA-24-a), one of the main active triterpenes isolated from the well-known medicinal plant Alisma orientale (Sam.) Juz., exhibits multiple biological activities including hypolipidemic activity. However, its effect on lipid metabolism in adipocytes remains unclear. The present study aimed to clarify the effect of AA-24-a on adipocyte lipolysis and to determine its potential mechanism of action using 3 T3-L1 cells. METHODS: We assayed the release of glycerol into culture medium of 3 T3-L1 cells under treatment with AA-24-a. Protein and mRNA expression and phosphorylation levels of the main lipases and kinases involved in lipolysis regulation were determined by quantitative polymerase chain reaction and western blotting. Specific inhibitors of protein kinase A (PKA; H89) and extracellular signal-regulated kinase (ERK; PD98059), which are key enzymes in relevant signaling pathways, were used to examine their roles in AA-24-a-stimulated lipolysis. RESULTS: AA-24-a significantly stimulated neutral lipolysis in fully differentiated adipocytes. To determine the underlying mechanism, we assessed the changes in mRNA and protein levels of key lipolysis-related genes in the presence or absence of H89 and PD98059. Both inhibitors reduced AA-24-a-induced lipolysis. Moreover, pretreatment with H89 attenuated AA-24-a-induced phosphorylation of hormone-sensitive lipase at Ser660, while pretreatment with PD98059 attenuated AA-24-a-induced downregulation of peroxisome proliferator-activated receptor-γ and perilipin A. CONCLUSIONS: Our results indicate that AA-24-a promoted neutral lipolysis in 3 T3-L1 adipocytes by activating PKA-mediated phosphorylation of hormone-sensitive lipase and ERK- mediated downregulation of expression of perilipin A.