Corilagin attenuates morphine-induced BV2 microglial activation and inflammation via regulating TLR2-mediated endoplasmic reticulum stress.

Morphine-induced microglia activation and neuroinflammation have been considered as the contributors of morphine tolerance. Corilagin (Cori) has been reported to exhibit strong anti-inflammatory property. The present study aims to investigate whether and how Cori alleviates morphine-induced neuroinflammation and microglia activation. Mouse BV-2 cells were exposed to different concentrations of Cori (0.1, 1 and 10 µM) prior to morphine stimulation (200 µM). Minocycline (10 µM) acted as the positive control. Cell viability was determined by CCK-8 assay and trypan blue assay. The levels of inflammatory cytokines were determined using ELISA. IBA-1 level was examined via immunofluorescence. TLR2 expression level was examined by quantitative real-time PCR and western blot. The expression levels of corresponding proteins were measured by western blot. It was found that Cori was non-toxic to BV-2 cells but greatly inhibited morphine-induced IBA-1 expression, overproduction of pro-inflammatory cytokines, activation of NLRP3 inflammasome and endoplasmic reticulum stress (ERS), and upregulation of COX-2 and iNOS. TLR2 was negatively regulated by Cori, and could promote the activation of ERS. A high affinity between Cori and TLR2 protein was confirmed via Molecular docking investigation. Moreover, TLR2 overexpression or tunicamycin (TM), an agonist of ERS, partly abolished the inhibitory effects of Cori on morphine-induced alternations on neuroinflammation and microglial activation in BV-2 cells as above. In summary, our study suggested that Cori effectively alleviated morphine-induced neuroinflammation and microglia activation through inhibiting TLR2-mediated ERS in BV-2 cells, providing a novel potential drug to overcome morphine tolerance.

In Situ Membrane Biotinylation Enables the Direct Labeling and Accurate Kinetic Analysis of Small Extracellular Vesicles in Circulation.

Circulating small extracellular vesicles (sEVs) are naturally occurring nanosized membrane vesicles that convey bioactive molecules between cells. Conventionally, to evaluate their behaviors in vivo, circulating sEVs have to be isolated from the bloodstream, then labeled with imaging materials in vitro, and finally injected back into the circulation of animals for subsequent detection. The tedious isolation-labeling-reinfusion procedures might have an undesirable influence on the natural properties of circulating sEVs, thereby changing their behaviors and the detected kinetics in vivo. Herein, we proposed an in situ biotinylation strategy to directly label circulating sEVs with intravenously injected DSPE-PEG-Biotin, aiming to evaluate the in vivo kinetics of circulating sEVs more biofriendly and accurately. Such an analysis strategy is free of isolation-labeling-reinfusion procedures and has no unfavorable influence on the natural behaviors of sEVs. The results showed that the lifetime of generic circulating sEVs in mice was around 3 days. Furthermore, we, for the first time, revealed the distinct in vivo kinetics of circulating sEV subpopulations with different cell sources, among which erythrocyte-derived sEVs showed the longest lifespan. Moreover, compared with circulating sEVs in situ or used as autograft, circulating sEVs used as allograft had the shortest lifetime. In addition, the in situ biotinylation strategy also provides a way for the enrichment of biotinylated circulating sEVs. In summary, this study provides a novel strategy for in situ labeling of circulating sEVs, which would facilitate the accurate characterization of their kinetics in vivo, thereby accelerating their future application as biomarkers and theranositic vectors.

Characterization of diacylglycerol acyltransferase 2 from Idesia polycarpa and function analysis.

Idesia polycarpa is an oil-producing tree native to China and Northeast Asia. The fruits of I. polycarpa which are named oil grape are unique in that they contain large amounts saturated and unsaturated lipids. Diacylglycerol acyltransferase 2 (DGAT2) is a key enzyme catalyzing the final step of triacylglyceride (TAG) synthesis. However, expression and bioinformatics of DGAT2 in I. polycarpa are still blank. In order to further understand the lipogenesis of oil grape, we contrasted seven various growth periods fruits from seed formation to seed maturation. Lipid accumulation rates and final lipid content were significantly different among the different periods. We cloned and characterized the DGAT2 gene from fruits of I. polycarpa. A partial fragment of 239 bp of IpDGAT2 was amplified by PCR. We cloned the open-reading frame (ORF) of IpDGAT2 by RACE technique. The ORF of IpDGAT2 contains 984 bp and encodes 327 amino acids. The qPCR analysis manifested that IpDGAT2 was expressed in all oil grape growing periods and expression was highest on September 20 (seed maturation). In I. polycarpa fruits the expression of IpDGAT2 was positively correlated with the lipid accumulation rates. Rhodotorula glutinis expression analysis showed that IpDGAT2 have a diacylglycerol acyltransferase bio-functional. Heterologous expression of the 35S::IpDGAT2 in Arabidopsis thaliana confirmed that the isolated IpDGAT2 could catalyze lipid synthesis. The lipid content increased by 40 % in transgenic plants relative to the control. which suggests that high lipid content fruits can be created by the overexpression of IpDGAT2 in I. polycarpa.

Magnetic Chip Based Extracorporeal Circulation: A New Tool for Circulating Tumor Cell in Vivo Detection.

In vivo detection of circulating tumor cells (CTCs) which inspect all of the circulating blood in body seems to have more advantages on cell capture, especially in earlier cancer diagnosis. Herein, based on in vivo microfluidic chip detection system (IV-chip-system), an extracorporeal circulation was constructed to effectively detect and monitor CTCs in vivo. Combined with microfluidic chip and immunomagnetic nanosphere (IMN), this system not only acts as a window for CTC monitoring but also serves as a collector for further cancer diagnosis and research on CTCs. Compared with the current in vivo detection method, this system can capture and detect CTCs in the bloodstream without any pretreatments, and it also has a higher CTC capture efficiency. It is worth mentioning that this system is stable and biocompatible without any irreversible damage to living animals. Taking use of this system, the mimicked CTC cleanup process in the blood vessel is monitored, which may open new insights in cancer research and early cancer diagnosis.

Insecticide residues in head lettuce, cabbage, Chinese cabbage, and broccoli grown in fields.

The residues of four insecticides belonging to different families were studied on head lettuce (Lactuca sativa L. var. capitata L.), cabbage (Brassica oleracea Linn. var. capitata DC.), Chinese cabbage (Brassica pekinensis Skeels), and broccoli (Brassica oleracea var. italica) after pesticide application. To reduce application variability, a tank mix of acetamiprid 20% SP, chlorpyrifos 22.5% EC, deltamethrin 2.4% SC, and methomyl 40% SP was applied at recommended and double doses. Initial deposits of all pesticides on head lettuce were higher than those of the other three crops. The residues of chlorpyrifos and deltamethrin were higher than the maximum residue limits (MRLs) at recommended preharvest intervals (PHIs) on head lettuce and Chinese broccoli treated with higher doses. The residues of methomyl on head lettuce also showed the same phenomenon.

Low-dose whole organ CT perfusion of the pancreas: preliminary study.

OBJECTIVES: To investigate the feasibility of low-dose whole pancreas CT perfusion in the clinical practice. METHODS: Sixty-one patients suspected pancreatic disease underwent low-dose whole pancreas CT perfusion scan (by body weight, group A: 70 kV, 120 mAs; group B: 80 kV, 100 mAs) and the individualized pancreas scan. Forty-six patients were enrolled. Perfusion characteristics, such as, blood flow, blood volume and permeability, were analyzed. The effective radiation dose of the whole pancreas CT perfusion and the total CT scan protocol were recorded. CT findings were histologically confirmed by surgical intervention or diagnostic puncture. RESULTS: Of the 46 cases, 33 were pancreatic adenocarcinoma, 5 were solid-pseudo-papillary tumors of pancreas, 8 cases of pancreatic endocrine tumors on the perfusion study. There was significant interobserver agreement on the measurement of normal pancreatic CT perfusion parameters of group A (n = 28)and group B (n = 18), respectively (p > 0.05). For the normal pancreas, there was no significant difference on CT perfusion parameters between group A and group B (p > 0.05). There were significant differences on blood flow as well as blood volume between the pancreatic adenocarcinomas and the normal pancreas (p < 0.001), whereas no difference on the permeability (p > 0.05). The time to peak of the normal pancreas is 28.94 ± 4.37 s (range from 24 to 38 s). Different pancreatic tumors had different types of time attenuation curve (TAC). TACs were different between pancreatic adenocarcinomas and normal pancreas. The effective radiation dose of the whole pancreas CT perfusion of Group A and Group B were 3.60 and 4.88 mSv (DLP 246 and 325 mGy cm), respectively, and the total radiation dose was around 8.01-16.22 mSv. CONCLUSIONS: Low-dose whole pancreatic CT perfusion can effectively reduce radiation dose, and provide the best phase for the individualized pancreas scan, which has great value in the clinical practice.

2009 pandemic characteristics and controlling experiences of influenza H1N1 virus 1 year after the inception in Hangzhou, China.

This paper described the epidemiology and controlling experiences of influenza H1N1 in Hangzhou in the past 1 year. A total of 2,078 cases confirmed by real-time quantitative PCR till March 31, 2010, were analyzed by SPSS 12.0 software. During the early pandemic stage, a patient must be tested for H1N1 nucleic acid once he/she had influenza-like symptoms with the epidemiological history in 7 days, and be diagnosed if it was positive. But in the pandemic peak, we made efforts to identify and save severe cases combined with pneumonia or hypoxemia or respiratory failure or septic shock or multiple organ dysfunctions and failure. In general, the prevalence was 2.77/100,000 (2,078/7,510,844); severity rate, 10.44% (217/2,078); fatality rate, 0.48% (10/2,078). The carrier and secondary attack rate were 9.52% (58/612) and 8.66% (53/612), respectively. About 50% of serious cases and 100% of deaths had the basic underlying diseases: cardiovascular diseases, 13.66% (25/217); chronic lung disease, 12.02% (22/217); pregnant, 7.1% (13/217). Of all cases aged from 1 month to 89 years, 52.99% (1,435/2,708) were in the 10-29 years, with most of them distributed in downtown area. The timeline showed two epidemic peaks occurred in September and November 2009, respectively. Furthermore, the hemagglutinin gene remained 99% identical with the American and vaccine strains, but only 70% with the 1947-2008 Chinese strains. In conclusion, Hangzhou pandemic influenza H1N1 was caused by the highly conserved virus, with low prevalence, transmission, and mortality, because we took efficient controlling methods.

An initial qualitative study of dual-energy CT in the knee ligaments.

OBJECTIVE: To study the clinical application of dual-energy CT (DECT) in the knee ligaments. METHODS: Twelve cases (24 knees) were scanned using dual-energy CT for the knee. Two- and three-dimensional images were used for display in all cases by means of multi-planar reformation (MPR) and volume rendering technique (VRT). All images were ranked by two radiologists according to the grade of knee ligament displayed, the definition of edge and attachment points of the knee ligament. RESULTS: The partial ligaments of 24 knees, such as the patellar ligament, fibular collateral ligament, anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) were clearly displayed; the tibial collateral ligament was not satisfactorily displayed. The transversal ligaments, such as lateral patellar retinaculum and medial patellar retinaculum, and the posterior ligament, such as oblique popliteal ligament could not be shown clearly. CONCLUSION: The dual-energy CT is a new and valuable tool to qualitatively display the main ligaments of the knee.

[Application of the trend-surface analysis for studying the geographical distribution of lung cancer mortality].

OBJECTIVE: To apply trend-surface analysis on lung cancer mortality in China. METHODS: To overview the theory and approach of trend-surface analysis and to fit the first to fifth order regression equations, where dependent variables were latitude and longitude, and the independent one was the standardized mortality ratio (SMR) of lung cancer for various provinces. Data was from the second mortality survey in the whole country, then proper regression equation was chosen. RESULTS: Fifth-order regression equation was established for mortality of lung cancer in males with its fit goodness as 76.06%. The third-order regression equation was established for mortality of lung cancer in females with its fit goodness as 89.77%. There were positive residuals in Shanghai, Hubei and Tianjin, while the negative residuals were in Zhejiang, Anhui, Henan, Beijing and Guangxi for males. There were positive residuals in Tianjin, Hubei and Guangdong, while negative residuals appeared in Zhejiang, Shanxi, Guizhou, Hainan, Henan, Jilin and Neimenggu for females. CONCLUSION: The geographic distribution trend of lung cancer mortality between males and females appeared to be different. The mortality of lung cancer among males was high in the northeast provinces while the mortality of lung cancer for females was high in the southeast areas. In those areas where the residual values were anomalous, further studies need to be carried out.