Discovery of 5-(3,4-dihydroxybenzylidene)-1,3-dimethylpyrimidine- 2,4,6(1H,3H,5H)-trione as a novel and effective cardioprotective agent via dual anti-inflammatory and anti-oxidative activities.

Myocardial ischemia/reperfusion (MI/R) injury is still the huge unmet medical need without effective therapy in clinic. It is critical to develop pharmacological intervention to scavenge ROS and inhibit NLRP3 activation to have a double benefit against MI/R injury. Cinnamamide derivatives have been demonstrated to possess anti-oxidative and anti-inflammatory activities. Previously, we have reported that a cinnamamide derivative 2 exerts excellent cardioprotective effect via mediation of intracellular oxidative stress via Nrf2 up-regulation against MI/R. In the present study, seventeen compounds have been optimized using cinnamamide-barbiturate hybrid 2 as the lead compound and their cardioprotective activities against MI/R were further determined in vitro and in vivo. Among them, compound 7 showed the most potent cardioprotective effect and low cytotoxicity. While cardiomyocytes were invased by hydrogen peroxide, compound 7 exhibited more excellent cardioprotective effect than that of luteolin and metoprolol, the positive control employed in the present study, as demonstrated by dramatically elevated cell survival rate and decreased LDH leakage rate. Moreover, compound 7 markedly inhibited cardiac expressions of inflammasome activation and pro-inflammatory cytokines release (i.e. NLRP3, IL-1ß, IL-18), simultaneouly increasing endogenous antioxidative proteins (i.e. Nrf2, HO-1 and SOD) in vitro. In the rat MI/R model, compound 7 pretreatment profoundly reduced cardiac infarct size in MI/R rats and reversed abnormal changes in myocardial enzymes and lipid peroxidation levels in heart tissues. Mechanistically, compound 7 revealed significant cardioprotective effects by inhibiting NLRP3 and its downstream inflammatory chemokine IL-1ß, as well as up-regulating Nrf2 in vivo. Furthermore, at the active site of the co-crystal of NLRP3 and Nrf2, compound 7 exhibited higher binding force in the molecular docking study, which was consistent with the in vitro results. Therefore, compound 7 is expected to be a potential cardioprotective agent possessing dual anti-inflammatory and anti-oxidative activities. Our work provides an important therapeutic strategy for the treatment of ischemic-reperfused heart disease.

Discovery of cinnamamide-barbiturate hybrids as a novel class of Nrf2 activator against myocardial ischemia/reperfusion injury.

Myocardial ischemia/reperfusion (MI/R) has been a challenge for global public health. Activation of nuclear factor erythroid-2-related factor 2 (Nrf2) signaling could attenuate MI/R injury by maintaining cell redox balance and reducing oxidative damage. Cinnamamide derivatives have been proven to be a class of potential Nrf2 activators and cardioprotective agents. The development of novel cinnamamide derivatives to combat oxidative stress in cardiomyocytes is highly desirable. In this study, twenty-three cinnamamide-barbiturate hybrids were studied. Cell-based assays showed that most of the compounds exhibited excellent protective activity against H2O2-induced oxidative injury in H9c2 cells. Notably, compound 7w, which had the highest activity and low cytotoxicity, was demonstrated to remarkably reduce intracellular ROS accumulation by activating the mRNA expression of Nrf2 and its downstream antioxidant gene HO-1, indicating a novel promising antioxidant and Nrf2 activator. The probable binding mode between protein Keap1 and compound 7w was also studied via molecule docking. Furthermore, we found that the administration of compound 7w could significantly reduce the cardiac infarct size and improve the cardiac function against MI/R injury in rats, as well as decrease cardiac oxidative stress. Taken together, we report, for the first time, that cinnamamide-barbiturate hybrids are a novel class of potential cardioprotective agents. The excellent cardioprotective action of such compounds rely on enhancing the endogenous antioxidative system by upregulating the Nrf2 signaling pathway in vitro and in vivo against MI/R damage. These findings provide a new perspective for designing cinnamamide-barbiturate hybrids as a novel class of Nrf2 activator against cardiovascular diseases.

Adipocyte-secreted microvesicle-derived miR-148a regulates adipogenic and osteogenic differentiation by targeting Wnt5a/Ror2 pathway.

AIMS: Adipocyte-secreted microvesicles (MVs)-derived microRNAs (miRNAs) are relevant to adipogenic and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in osteonecrosis of the femoral head (ONFH). Our aims are to investigate the mechanism of adipocyte-derived MVs-miR-148a in ONFH. MATERIALS AND METHODS: Adipocyte-derived MVs were identified via transmission electron microscopy and specific markers expression. The adipogenic and osteogenic differentiation were investigated by Oil-Red O staining, alkaline phosphatase (ALP) activity, Alizarin Red S (ARS) staining and osteogenic or adipogenic factors levels. Genes and proteins expression were detected by using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The relationship between miR-148a and Wnt5a was tested via dual-luciferase reporter analysis. The adipogenic differentiation and osteogenic differentiation in methylprednisolone (MPS)-induced ONFH rat model were assessed via hematoxylin-eosin (HE) staining, and immunohistochemical staining of collagen I (COL I). KEY FINDINGS: Adipocyte-derived MVs promoted adipogenic differentiation via increasing Oil-Red O staining positive cells, adiponectin (Adipoq), acid-binding protein 2 (aP2) and peroxisome proliferator-activated receptor γ (PPAR-γ) levels, and repressed osteogenic differentiation of BMSCs via decreasing ARS staining positive cells, ALP, Runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN) levels. MiR-148a was present in adipocyte-derived MVs, and miR-148a knockdown inhibited adipogenic differentiation and promoted osteogenic differentiation. Furthermore, Wnt5a expression was regulated by miR-148a. MiR-148a overexpression facilitated adipogenic differentiation and suppressed osteogenic differentiation via regulating the Wnt5a/Ror2 pathway. Adipocyte-derived MVs promoted adipogenic differentiation and inhibited osteogenic differentiation in MPS-induced ONFH rat model. SIGNIFICANCE: Adipocyte-derived MVs-miR-148a promoted adipogenic differentiation and suppressed osteogenic differentiation via targeting the Wnt5a/Ror2 pathway.

Modelling and assessing the effects of medical resources on transmission of novel coronavirus (COVID-19) in Wuhan, China.

The coronavirus disease 2019 (COVID-2019), a newly emerging disease in China, posed a public health emergency of China. Wuhan is the most serious affected city. Some measures have been taken to control the transmission of COVID-19. From Jan. 23rd, 2020, gradually increasing medical resources (such as health workforce, protective clothing, essential medicines) were sent to Wuhan from other provinces, and the government has established the hospitals to quarantine and treat infected individuals. Under the condition of sufficient medical resources in Wuhan, late-stage of epidemic showed a downward trend. Assessing the effectiveness of medical resources is of great significance for the future response to similar disease. Based on the transmission mechanisms of COVID-19 and epidemic characteristics of Wuhan, by using time-dependent rates for some parameters, we establish a dynamical model to reflect the changes of medical resources on transmission of COVID-19 in Wuhan. Our model is applied to simulate the reported data on cumulative and new confirmed cases in Wuhan from Jan. 23rd to Mar. 6th, 2020. We estimate the basic reproduction number R0 = 2.71, which determines whether the disease will eventually die out or not under the absence of effective control measures. Moreover, we calculate the effective daily reproduction ratio Re(t), which is used to measure the 'daily reproduction number'. We obtain that Re(t) drops less than 1 since Feb. 8th. Our results show that delayed opening the 'Fire God Hill' hospital will greatly increase the magnitude of the outbreak. This shows that the government's timely establishment of hospitals and effective quarantine via quick detection prevent a larger outbreak.

Overexpression of Tripartite Motif-Containing 48 (TRIM48) Inhibits Growth of Human Glioblastoma Cells by Suppressing Extracellular Signal Regulated Kinase 1/2 (ERK1/2) Pathway.

BACKGROUND Herein, we found that tripartite motif-containing 48 (TRIM48) was reduced in human glioblastoma (GBM) cell lines. We investigated whether and how TRIM48 functions in human GBM in vitro. MATERIAL AND METHODS Human GBM cells (U87 MG and U138 MG) were infected with lentivirus to overexpress TRIM48, and 1 human GBM cell line (T98G) was infected with siRNAs to knock down TRIM48 expression. Techniques used included cell proliferation assay, measured by CCK-8 and BrdU-ELISA method, and cell cycle assay, determined using flow cytometry. Curcumin, a specific activator of extracellular signal regulated kinases (ERK1/2), or PD98059, a specific inhibitor of ERK1/2, was used to activate or block the ERK1/2 pathway, respectively. Expression of phosphorylated (p)-ERK1/2, and its downstream targets (Cyclin D1) were measured to assess the mechanism. RESULTS Our data suggest that overexpression of TRIM48 reduces the viability of U87 MG and U138 MG and leads to cell cycle arrest (in G0-G1 phase), which is associated with blockade of the ERK1/2 pathway and reduction of Cyclin D1. In contrast, knockdown of TRIM48 resulted in the opposite effects. Interestingly, the inhibitory effect of TRIM48 overexpression on human GBM cell growth and the inactivation of ERK1/2 were significantly alleviated with additional curcumin treatment, while it the promoted the effect of siTRIM48 on human GBM cell growth, and the activation of ERK1/2 was significantly alleviated with additional PD98059 treatment. CONCLUSIONS TRIM48 suppressed the growth of human GBM cell via the prevention of ERK1/2 activation.

[Spatial-temporal pattern of sustainable intensification of agricultural land-use in Shandong Province, China.] / å±±ä¸œçœå†œåœ°åˆ©ç”¨å¯æŒç»­é›†çº¦åŒ–çš„æ—¶ç©ºæ ¼å±€.

Under the theoretical framework of sustainable intensification of agricultural land-use (SIALU), We used material flow analysis (MFA) method to establish evaluation index system for SIALU by utilizing data in 2000, 2005, 2010 and 2015 to quantify the level of SIALU of 17 cities in Shandong Province, and analyzed the variation in input-output of resources factors of agricultural land, spatial distribution of resource productivity and environmental economic efficiency, in order to reveal spatial-temporal differentiation of SIALU. Results showed that the direct material input to agricultural lands decreased, whereas hidden flow, stock and pollutant emissions increased gradually from 2000 to 2015. The material productivity of all cities in the province showed that the coastal areas in the peninsula were relatively lower than the southern region, and the level of material productivity in the northwest region was relatively higher. Environmental economic efficiency was gradually enhanced, and the western region was relatively higher than coastal area of the peninsula. During the period examined here, the spatial pattern of SIALU of various cities showed clustered distribution change, with the western region tending to gradually increase and the eastern region tending to gradually reduce. The dynamics of SIALU among different regions were divided into six grades: Northwestern Shandong > Northern Shandong > Southwestern Shandong > Southern Shandong > Central Shandong > Coastal areas of Shandong Peninsula.

Early postoperative radiotherapy is associated with improved outcomes over late postoperative radiotherapy in the management of completely resected (R0) Stage IIIA-N2 non-small cell lung cancer.

AIMS: The aim of this study was to evaluate the ideal timing of PORT in the management of completely resected (R0) Stage IIIA-N2 NSCLC. PATIENTS AND METHODS: Between January 2008 and December 2015, patients with known histologies of pathologic Stage IIIA-N2 NSCLC who underwent R0 resection and received PORT concurrent with or prior to two sequential cycles of chemotherapy ("early PORT") or with PORT administered after two cycles of chemotherapy ("late PORT") at multiple hospitals. The primary endpoint was OS; secondary end points included pattern of the first failure, LRRFS, and DMFS. Kaplan-Meier OS, LRRFS, and DMFS curves were compared with the log-rank test. Cox regression analysis was used to determine prognosticators for OS, LRRFS, and DMFS. RESULTS: Of 112 included patients, 41 (36.6%) and 71 (63.4%) patients received early PORT and late PORT, respectively. The median OS, LRRFS, and DMFS were longer for those who received early PORT than for those who received late PORT at the median follow-up of 29.6 months (all p < 0.05). Uni- and multi-variate analyses showed that number of POCT cycles and the combination schedule of PORT and POCT were independent prognostic factors for OS, LRRFS, and DMFS. CONCLUSIONS: Early PORT is associated with improved outcomes in pathologic Stage IIIA-N2 R0 NSCLC patients.

Prognostic significance of the pretreatment serum gamma-glutamyltransferase levels in Chinese patients with non-metastatic cervical cancer.

This study was performed to evaluate the prognostic significance of the pretreatment serum gamma-glutamyltransferase (GGT) levels in a Chinese cohort of patients with early-stage or locally advanced cervical cancer. The pretreatment serum GGT levels were examined in 290 cervical cancer patients with stage I-III disease and 230 healthy controls selected from a cancer-free population in the same region. Patients were assigned to normal or high-risk GGT groups, as previously described, and the GGT levels were correlated to clinicopathologic parameters and survival data. The GGT levels in cervical cancer patients were significantly higher than those in healthy controls (35.6 ± 29.1 vs. 24.1 ± 14.7 U/L, P < 0.001). In addition, the pretreatment serum GGT levels were associated with the histology type (P = 0.023), lymph node involvement (P = 0.040), stage (P = 0.029), recurrence (P = 0.015) and death (P = 0.005), but not with age (P = 0.432), tumor size (P = 0.067) or degree of differentiation (P = 0.901). Moreover, univariate survival analysis revealed that patients with high GGT levels tended to have poorer disease-free survival (DFS) [hazard ratio (HR), 1.721; 95% confidence interval (CI), 1.189-2.491; P = 0.004] and overall survival (OS) (HR, 1.929; 95% CI, 1.294-2.876; P = 0.001) compared to those with normal GGT levels. However, a multivariate Cox-regression model did not support these data (HR, 1.373; 95% CI, 0.925-2.039; P = 0.116 for DFS and HR, 1.357; 95% CI, 0.887-2.078; P = 0.160 for OS, respectively) after adjusting for other confounding variables. High pretreatment serum GGT was associated with more advanced tumor behavior, but could not serve as an independent prognostic indicator in patients with early-stage or locally advanced cervical cancer.

Necroptosis in tumorigenesis, activation of anti-tumor immunity, and cancer therapy.

While the mechanisms underlying apoptosis and autophagy have been well characterized over recent decades, another regulated cell death event, necroptosis, remains poorly understood. Elucidating the signaling networks involved in the regulation of necroptosis may allow this form of regulated cell death to be exploited for diagnosis and treatment of cancer, and will contribute to the understanding of the complex tumor microenvironment. In this review, we have summarized the mechanisms and regulation of necroptosis, the converging and diverging features of necroptosis in tumorigenesis, activation of anti-tumor immunity, and cancer therapy, as well as attempts to exploit this newly gained knowledge to provide therapeutics for cancer.

[Geo-spectrum characteristics of land use change in Jiangsu Province, China.] / 江苏省土地利用变化的图谱特征.

This paper studied the spatial-temporal characteristics and rules of land use change in Jiangsu Province using theories and methods of geo-spectrum. Based on the land use data translated from remote sensing images of 1990, 2000 and 2010, we synthesized the geo-spectrum of the mode of arable land use change and that of land use change in two corresponding phases, 1990-2000 and 2000-2010, in Jiangsu using ArcGIS 10.0. The results showed that in the phase of 1990-2000, the major characteristics of land use change were swaps between the geo-spectrum unit of arable land and urban-rural construction land, arable land and water body, and arable land and grassland. Specifically, the patterns of "arable land → urban-rural construction land" and "arable land→ water body" were highly significant. We also found the reduction of arable land area and the concentration of its spatial distribution. In the phase of 2000-2010, the "arable land → urban-rural construction land" pattern was still the most salient characteristic. In addition, the patterns of "grassland → water body" and "urban-rural construction land → water body" became more spatially concentrated and tended to expand. Compared with the previous phase, the area of the land use in the phase of 2000-2010 had been changed expanded and became more scattered. Overall, the geo-spectrum of arable land use change in Jiangsu was mainly shaped by the anaphase change type and partially by the prophase change type, with a tiny influence of the repeated and continuous change.

Phenanthrenes, 9,10-dihydrophenanthrenes, bibenzyls with their derivatives, and malate or tartrate benzyl ester glucosides from tubers of Cremastra appendiculata.

Eleven previously unknown compounds and 23 known compounds, including 20 phenanthrene or 9,10-dihydrophenanthrene derivatives, five bibenzyls, seven malate or tartrate benzyl ester glucosides, adenosine and gastrodin were isolated from tubers of Cremastra appendiculata. Among the obtained compounds, two are the first isolated dimers with one phenanthrene or bibenzyl unit connected to C-3 of 2,3,4,5-tetrahydro-phenanthro[2,1-b]furan moiety. In addition, 33 of these compounds were evaluated in vitro for their cytotoxic activity against two cancer cell lines. Among the compounds examined, one compound showed moderate cytotoxic activity, while five showed weak cytotoxic activity against the A549 cell line.

Simultaneous determination of atractylenolide II and atractylenolide III by liquid chromatography-tandem mass spectrometry in rat plasma and its application in a pharmacokinetic study after oral administration of Atractylodes Macrocephala Rhizoma extract.

Atractylenolide II (AII) and atractylenolide III (AIII) are the major active components in Atractylodes Macrocephala Rhizoma (AMR). In this study, a sensitive, rapid and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of AII and AIII in rat plasma using loliolide as internal standard (IS). After protein precipitation with ethyl acetate, the analytes were injected into an LC-MS/MS system for quantification. Chromatography was performed using a C(18) column, eluting with water and acetonitrile (45:55, v/v) at 0.2 mL/min. All analytes including IS were monitored under positive ionization conditions by multiple reaction monitoring with an electrospray ionization source. The validated method was successfully applied to the pharmacokinetic study of AII and AIII in rat plasma after oral administration of AMR extract. The results provided a meaningful basis for evaluating the clinical applications of traditional Chinese medicine.

Chemical constituents of Spatholobus suberectus.

AIM: To investigate chemical constituents of Spatholobus suberectus Dunn. METHODS: Isolation and purification were carried out by column chromatographic methods. Compounds were characterized based on their physical characteristics and spectra data. RESULTS: Seventeen compounds were isolated from ethanol extract of S. suberectus. The structures were elucidated as prestegane B (1), (2R, 3R)-buteaspermanol (2), (+)-medioresinol (3), (2R, 3R)-3,7-dihydroxyflavanone (4), benzeneethanol (5), 4, 7, 2'-trihydroxy-4'-methoxyisoflavanol (6), naringenin (7), blumenol A (8), protocatechuic acid ethyl ester (9), liquiritigenin (10), 7, 4'-dihydroxy-8-methoxy-isoflavone (11), 3, 5, 7, 3', 5'-pentahydroxyflavanone (12), protocatechuic acid (13), glycyroside (14), 8-methylretusin-7-O-ß-D-glucopyranoside (15), 3, 3', 4', 5, 6, 7, 8-heptahydroxyflavan (16), and dulcisflavan (17). CONCLUSION: All compounds are firstly isolated from the title plant and compounds 1, 3 were isolated from the Spatholobus genus for the first time.

Simultaneous determination of four sesquiterpenoids in Atractylodes Macrocephala Rhizoma by GC-FID: optimisation of an ultrasound-assisted extraction by central composite design.

INTRODUCTION: Atractylodes Macrocephala Rhizoma (AMR) is a traditional Chinese medicine containing several sesquiterpenoids with a series of effects. These bioactive compounds may be used as chemical markers for the quality control of AMR. It is necessary to optimise the extraction method and conditions in order to improve extraction productivity. OBJECTIVE: To develop a simple and effective method for the extraction of sesquiterpenoids from AMR and then to simultaneously determine four sesquiterpenoids, selina-4 (14), 7(11)-dien-8-one (SA), atractylenolide II (AII), atractylenolide III (AIII) and atractylenolide VII (AVII), in AMR. METHODOLOGY: Ultrasound-assisted extraction (UAE) was optimised by central composite design (CCD) to obtain the maximum efficiency. The gas chromatography method was validated and applied for the quantification of four sesquiterpenoids. RESULTS: The optimum values of factors were: particle size (120 mesh), extraction time (26 min), extraction temperature (39°C) and 31 mL of chloroform. The selectivity, linear range, limits of detection (LOD) and quantification (LOQ), accuracy, precision and repeatability of the method developed indicated its validity. The application of the method showed that the contents of four sesquiterpenoids in AMR were rather variable. CONCLUSION: The results indicated that the described GC method could be used for the quality control of AMR and its related preparations. Meanwhile, this research revealed that UAE under optimum conditions could be considered as a powerful tool for the extraction of phytochemicals from plants.