Comparative transcriptome analysis of unfractionated peripheral blood leukocytes after exercise in human.

Exercise has profound but variable effects on the immune system. However, only limited information exists about the changes of exercise-induced gene expression in whole immune cells. The aim of this study is to unravel the potential molecular changes of genes which are related to immunity after exercise. The raw expression data and corresponding clinical of GSE18966 were downloaded from Gene Expression Omnibus database. The differentially expressed genes between control group and treat groups were performed by in-house developed perl scripts. A total of 83 differentially expressed genes (DEGs) (|log2 FC|> 1, FDR < 0.05) were identified between control and treat group 1 (0 h after exercise), 128 DEGs (|log2 FC|> 1, FDR < 0.05) between control and treat group 2 (4 h after exercise), and there was no significant difference between control and treat group 3 (20 h after exercise). Next, we identified 51 overlapping genes between treat group 1 (0 h after exercise) and treat group 2 (4 h after exercise) using Venn analysis. Protein-protein interaction (PPI) network was constructed by Cytoscape 3.7.2, and nine hub genes (S100A12, FCGR3B, FPR1, VNN2, AQP9, MMP9, OSM, NCF4, HP) were identified. Finally, 9 hub genes were identified as the potential biomarkers of exercise using validation set (GSE83578) verification analysis. These hub genes might serve as potential molecular targets of monitoring exercise and training processes in the further.

miR-96-5p enhances cell proliferation and invasion via targeted regulation of ZDHHC5 in gastric cancer.

OBJECTIVE: To explore the biological function and mechanism of miR-96-5p in gastric cancer. METHODS: The expression of differently expressed microRNAs (DEMs) related to gastric adenocarcinoma (GAC) prognosis was identified in GAC tumor samples and adjacent normal samples by qRT-PCR. A target gene miR-96-5p was selected using TargetScan, miRTarBase, miRDB databases. The combination of miR-96-5p and ZDHHC5 was verified by luciferase receptor assay. To further study the function and mechanism of miR-96-5p, we treated MGC-803 cells with miR-96-5p inhibitor and si-ZDHHC5, then detected cell viability, apoptosis, migration and invasion ability, as well as the expression of ZDHHC5, Bcl-2, Bax, cleaved caspase-3, cleaved caspase-9, and COX-2 by Western blot. RESULTS: Compared with adjacent normal samples, the levels of miR-96-5p, miR-222-5p, and miR-652-5p were remarkably increased, while miR-125-5p, miR-145-3p, and miR-379-3p were significantly reduced in GAC tumor samples (P<0.01), which were consistent with bioinformatics analysis. Furthermore, ZDHHC5 was defined as a direct target gene of miR-96-5p. miR-96-5p silence significantly reduced cell viability, increased cell apoptosis, and suppressed cell migration and invasion, as well as inhibited the expression of Bcl-2 and COX-2 and promoted Bax, cleaved caspase-3 and cleaved caspase-9 level in MGC-803 cells (P<0.01). Notably, ZDHHC5 silence reversed the inhibiting effects of miR-96-5p on MGC-803 cells growth and metastasis Conclusion: Our findings identified six microRNAs (miRNAs; miR-96-5p, miR-222-5p, miR-652-5p, miR-125-5p, miR-145-3p, and miR-379-3p) related to GAC prognosis, and suggested that down-regulated miR-96-5p might inhibit tumor cell growth and metastasis via increasing ZDHHC5 expression enhance MGC-803 cell apoptosis, as well as decrease MGC-803 cell metastasis.

Mechanistic Design of Chemically Diverse Polymers with Applications in Oral Drug Delivery.

Polymers play a key role in stabilizing amorphous drug formulations, a recent strategy employed to improve solubility and bioavailability of drugs delivered orally. However, the molecular mechanism of stabilization is unclear, therefore, the rational design of new crystallization-inhibiting excipients remains a substantial challenge. This article presents a combined experimental and computational approach to elucidate the molecular features that improve the effectiveness of cellulose polymers as solution crystallization inhibitors, a crucial first step toward their rational design. Polymers with chemically diverse substituents including carboxylic acids, esters, ethers, alcohols, amides, amines, and sulfides were synthesized. Measurements of nucleation induction times of the model drug, telaprevir, show that the only effective polymers contained carboxylate groups in combination with an optimal hydrocarbon chain length. Computational results indicate that polymer conformation as well as solvation free energy are important determinants of effectiveness at inhibiting crystallization and show that simulations are a promising predictive tool in the screening of polymers. This study suggests that polymers need to have an adequate hydrophilicity to promote solvation in an aqueous environment, and sufficient hydrophobic regions to drive interactions with the drug. Particularly, the right balance between key substituent groups and lengths of hydrocarbon side chains is needed to create effective materials.

Design, synthesis and antimicrobial activity of 6-N-substituted chitosan derivatives.

Three novel 6-N-substituted chitosan derivatives were designed and synthesised and characterized by FTIR and NMR. The degree of substitution was calculated by elemental analysis results. The antimicrobial activities of the target compounds were evaluated by twofold serial broth dilution method and poisoned food technique. The antifungal activities of 6-aminoethylamino-6-deoxy chitosan (3), 6-butylamino-6-deoxy chitosan (4) and 6-pyridyl-6-deoxy chitosan (5) were significantly increased against Rhizoctonia cerealis, Fusarium oxysporum and Botrytis cinerea, and the inhibition rate ranged from 22.48% to 63.56% at the concentration of 0.2mg/mL. The compound 3 had better antibacterial activities than chitosan, and the minimum inhibition concentration of which ranged between 6.25 and 25mg/L against gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis and Bacillus anthracis) and gram-negative bacteria (Escherichia coli, Salmonella typhi). The antibacterial activities of 6-N-substituted chitosan tended to increase with the increase of the number of -NH2 group.

Hydroboration-oxidation: A chemoselective route to cellulose ω-hydroxyalkanoate esters.

We describe the first synthesis of hydroxy-functionalized polysaccharide esters via chemoselective olefin hydroboration-oxidation in the presence of ester groups. Cellulose esters with terminally olefinic side chains were first synthesized by esterification of commercially available cellulose esters (e.g., cellulose acetate) with undec-10-enoyl chloride or pent-4-enoyl chloride. Subsequent two-step, one-pot hydroboration-oxidation reactions of the cellulose esters were performed, using 9-borabicyclo[3.3.1]nonane as hydroboration agent, followed by oxidizing the intermediate borane to a hydroxyl group using mildly alkaline H2O2. Sodium acetate was used as a weak base to catalyze the oxidation, thereby minimizing undesired ester hydrolysis. Characterization methods including FTIR, (1)H, and (13)C NMR proved the selectivity of the hydroboration-oxidation pathway, providing a family of novel cellulose ω-hydroxyalkanoyl esters that were previously difficult to access.

Synthesis of amide-functionalized cellulose esters by olefin cross-metathesis.

Cellulose esters with amide functionalities were synthesized by cross-metathesis (CM) reaction of terminally olefinic esters with different acrylamides, catalyzed by Hoveyda-Grubbs 2nd generation catalyst. Chelation by amides of the catalyst ruthenium center caused low conversions using conventional solvents. The effects of both solvent and structure of acrylamide on reaction conversion were investigated. While the inherent tendency of acrylamides to chelate Ru is governed by the acrylamide N-substituents, employing acetic acid as a solvent significantly improved the conversion of certain acrylamides, from 50% to up to 99%. Homogeneous hydrogenation using p-toluenesulfonyl hydrazide successfully eliminated the α,ß-unsaturation of the CM products to give stable amide-functionalized cellulose esters. The amide-functionalized product showed higher Tg than its starting terminally olefinic counterpart, which may have resulted from strong hydrogen bonding interactions of the amide functional groups.

A descriptive analysis of injury triage, surge of medical demand, and resource use in an university hospital after 8.12 Tianjin Port Explosion, China.

OBJECTIVE: The 8.12 Tianjin Port Explosion in 2015 caused heavy casualties. Pingjin Hospital, an affiliated college hospital in Tianjin, China participated in the rescue activities. This study aims to analyze the emergency medical response to this event and share experience with trauma physicians to optimize the use of medical resource and reduce mortality of critical patients. METHODS: As a trauma centre at the accident city, our hospital treated 298 patients. We retrospectively analyzed the data of emergency medical response, including injury triage, injury type, ICU patient flow, and medical resource use. RESULTS: There were totally 165 deaths, 8 missing, and 797 non-fatal injuries in this explosion. Our hospital treated 298 casualties in two surges of medical demand. The first one appeared at 1 h after explosion when 147 wounded were received and the second one at 4 h when 31 seriously injured patients were received, among whom 29 were transferred from Tianjin Emergency Center which was responsible for the scene injury triage. After reexamination and triage, only 11 cases were defined as critical ill patients. The over-triage rate reached as high as 62.07%. Seventeen patients underwent surgery and 17 patients were admitted to the intensive care unit. CONCLUSIONS: The present pre-hospital system is incomplete and may induce two surges of medical demand. The first one has a much larger number of casualties than predicted but the injury level is mild; while the second one has less wounded but almost all of them are critical patients. The over-triage rate is high. The hospital emergency response can be improved by an effective re-triage and implementation of a hospital-wide damage control.

Olefin cross-metathesis as a source of polysaccharide derivatives: cellulose ω-carboxyalkanoates.

Cross-metathesis has been shown for the first time to be a useful method for the synthesis of polysaccharide derivatives, focusing herein on preparation of cellulose ω-carboxyalkanoates. Commercially available cellulose esters were first acylated with 10-undecenoyl chloride, providing esters with olefin-terminated side chains. Subsequent cross-metathesis of these terminal olefin moieties with acrylic acid was performed in solvents including acrylic acid, THF, and CH2Cl2. Complete conversion to discrete, soluble cross-metathesis products was achieved by using the Hoveyda-Grubbs second generation ruthenium catalyst and an excess of acrylic acid. Oligomerization during storage, caused by a free radical mechanism, was observed and successfully suppressed by the addition of a free radical scavenger (BHT). Furthermore, the cross-metathesis products exhibited glass transition temperatures (Tg) that were at least 50 °C higher than ambient temperature, supporting the potential for application of these polymers as amorphous solid dispersion matrices for enhancing drug aqueous solubility.

Isolation and in vitro partial characterization of hemolytic proteins from the nematocyst venom of the jellyfish Stomolophus meleagris.

Jellyfish venom contains various toxins and can cause itching, edema, muscle aches, shortness of breath, blood pressure depression, shock or even death after being stung. Hemolytic protein is one of the most hazardous components in the venom. The present study investigated the hemolytic activity of the nematocyst venom from jellyfish Stomolophus meleagris. Anion exchange chromatography, DEAE Sepharose Fast Flow, and gel filtration chromatography, Superdex200 had been employed to isolate hemolytic proteins from the nematocyst venom of jellyfish S. meleagris. Hemolysis of chicken red blood cells was used to quantify hemolytic potency of crude nematocyst venom and chromatography fractions during the purification process. Native-PAGE profile displayed one protein band in the purified hemolytic protein (SmTX); however, two protein bands with apparent molecular weights of ≈ 45 kDa and 52 kDa were observed in the reducing SDS-PAGE analysis. Approximately 70 µg/mL of SmTX caused 50% hemolysis (HU50) of the erythrocyte suspension. The hemolytic activity of SmTX was shown to be temperature and pH dependent, with the optimum temperature and pH being 37°C and pH 5.0. The present study is the first report of isolation and partial characterization of hemolytic proteins from the nematocyst venom of the jellyfish S. meleagris. The mechanism of the hemolytic activity of SmTX is not clear and deserves further investigation.

Microwave-assisted degradation of chitosan for a possible use in inhibiting crop pathogenic fungi.

Degradation of chitosan by H(2)O(2) under microwave irradiation was investigated. The oxidative degradation of chitosan was highly accelerated by microwave irradiation under the condition of low temperature and low concentration of H(2)O(2). The degraded chitosans with low molecular weight (M(w)) were characterized by gel permeation chromatography, Fourier-transform infrared spectroscopy, ultraviolet-visible spectroscopy, X-ray diffraction and elemental analysis. The decrease of M(w) led to transformation of crystal structure and increase of water solubility, whereas no significant chemical structure change in the backbone of chitosan was observed. Antifungal activities of chitosans with different M(w) against crop pathogenic fungi Phomopsis asparagi, Fusarium oxysoporum f. sp. Vasinfectum and Stemphylium solani were investigated at the concentrations of 100, 200 and 400 mg/L. All degraded chitosans with low M(w) exhibited enhanced antifungal activity compared with original chitosan and the chitosan of 41.2 kDa showed the highest activity. At 400 mg/L, the chitosan of 41.2 kDa inhibited growth of P. asparagi at 89.3%, stronger than polyoxin and triadimefon, the inhibitory effects of which were found to be 55.5% and 68.5%. All the results indicated that oxidative degradation under microwave irradiation was a promising technique for large-scale production of low M(w) chitosan for use in crop protection.

Application of nanoLC-MS/MS to the shotgun proteomic analysis of the nematocyst proteins from jellyfish Stomolophus meleagris.

The nematocyst proteins of jellyfish Stomolophus meleagris, a complicated mixture, contain many important bioactive molecules. In present study, to gain comprehensive insight into the protein component and search some novel bioactive molecules in the nematocyst proteins, shotgun proteomic analysis of the nematocyst proteins was carried out by nano liquid chromatography tandem mass spectrometry (nanoLC-MS/MS) for the first time. Digested peptides of the nematocyst proteins were analyzed by nanoLC-MS/MS and all MS/MS spectra were then automatically searched by the SEQUEST program. A total of 181 proteins had been identified, with the molecular weight ranging from 5268.06 to 843,487.57 and the pI from 4.49 to 11.39. Bioinformatic analysis was also applied to better understand the identified proteins. In the gene ontology (GO) annotation, all the identified proteins were classified into 13, 9 and 7 groups according to biological process, cellular component and molecular function, respectively. Pathways analysis of the identified proteins was conducted with 33 corresponding pathways found. On the basis of pathways analysis, we also constructed the gene network to analyze the relationship of those genes each other, which contained enzyme-enzyme relation, protein-protein interaction and gene expression interaction.

Isolation, identification and characterization of a novel antioxidant protein from the nematocyst of the jellyfish Stomolophus meleagris.

Reactive oxygen species (ROS) can damage the lipids, proteins and DNA when produced excessively in cells. Here, we describe the isolation and identification of a novel antioxidant protein named SmP90 from the nematocyst of jellyfish Stomolophus meleagris by 50% ammonium sulfate precipitation and gel filtration chromatography, superdex75. HPLC and SDS-PAGE analysis revealed >95% purity of SmP90 with apparent molecular weight of 90 kDa, approximately. The identification of SmP90 was confirmed by both N-terminal amino acids sequencing, with the sequences of NLDTPYCFYSGDYGG, and peptide mass fingerprint (PMF) analysis by MALDI-TOF-MS. However, no known protein had been completely matched in the database, which indicated that SmP90 might be a novel protein. The antioxidant assay result showed that it had strong superoxide anion radical-scavenging activity with the half-scavenging concentration (EC(50)) of about 16 µg/mL. Therefore, the present study is the first time to demonstrate a high efficient method of isolating a novel antioxidant protein from the nematocyst of jellyfish S. meleagris.

Molecular weight and pH effects of aminoethyl modified chitosan on antibacterial activity in vitro.

Aminoethyl modified chitosan derivatives (AEMCSs) with different molecular weight (Mw) were synthesized by grafting aminoethyl group on different molecular weight chitosans and chitooligosaccharide. FTIR, (1)H NMR, (13)C NMR, elemental analysis and potentiometric titration results showed that branched polyethylimine chitosan was synthesized. Clinical Laboratory Standard Institute (CLSI) protocols were used to determine MIC for Gram-negative strain of Escherichia coli under different pH. The antibacterial activity of the derivatives was significantly improved compared with original chitosans, with MIC values against E. coli varying from 4 to 64 µg/mL depending on different Mw and pH. High molecular weight seems to be in favor of stronger antibacterial activity. At pH 7.4, derivatives with Mw above 27 kDa exhibited equivalent antibacterial activity (16 µg/mL), while oligosaccharide chitosan derivative with lower Mw (~1.4 kDa) showed decreased MIC of 64 µg/mL. The effect of pH on antibacterial activity is more complicated. An optimal pH for HAEMCS was found around 6.5 to give MIC as low as 4 µg/mL, while higher or lower pH compromised the activity. Cell integrity assay and SEM images showed evident cell disruption, indicating membrane disruption may be one possible mechanism for antibacterial activity.

Synthesis and characterization of dithiocarbamate chitosan derivatives with enhanced antifungal activity.

In this study, ammonium dithiocarbamate chitosan (ADTCCS) and triethylene diamine dithiocarbamate chitosan (TEDADTCCS) derivatives were obtained respectively by mixing chitosan with carbon disulfide and ammonia (triethylenediamine). Their structures were confirmed by FT-IR, 1H NMR, XRD, DSC, SEM, and elemental analysis. Antifungal properties of them against the plant pathogenic fungi Fusarium oxysporum and Alternaria porri were investigated at concentrations ranged from 31.25 to 500 mg/L. The dithiocarbamate chitosan derivatives had enhanced antifungal activity compared with chitosan. Particularly, they showed obvious inhibitory effect on Fusarium oxysporum. At 500 mg/L, TEDADTCCS inhibited growth of F. oxysporum at 60.4%, stronger than polyoxin and triadimefon whose antifungal indexes were found to be 25.3% and 37.7%. The chitosan derivatives described here deserve further study for use in crop protection.