[The role of RUNX1 in the apoptosis of epithelial cells in nasal polyps].

Objective: To explore the expression of Runt-related transcription factor 1 (RUNX1) in nasal polyps (NPs) tissues and the potential role on apoptosis of primary human nasal epithelial cells (pHNECs) in NPs. Methods: The expression level of RUNX1 in NPs tissues was determined by Western blot (WB) and immunohistochemical staining (IHC). In vitro, TNF-α (20 ng/ml) was used to stimulate pHNECs to establish the apoptosis injury model. Hoechst staining was performed to observe pHNECs apoptosis by kit. Subsequently, quantitative real-time PCR (qRT-PCR) and WB were utilized to detect the expression of apoptosis-related proteins B-cell lymphoma-2 (BCL-2), BCL2-associated X (BAX) and cysteinyl aspartate specific proteinase-3 (Caspase-3) to assess the level of apoptosis. The plasmid of sh-RUNX1-6 was transfected into the pHNECs apoptosis model, then the effect of RUNX1 silence on apoptosis was evaluated by WB and flow cytometry. Statistical analysis was performed by the SPSS 19.0 and GraphPad Prism5 software. Results: The expression of RUNX1 in NPs tissue was significantly higher than that in inferior turbinates, and the difference was statistically significant (0.274±0.042 vs 0.110±0.027, t=9.675, P<0.05). Compared with the inferior turbinates, BAX and Caspase-3 expressions were increased whereas BCL-2 was decreased in NPs, and the differences were statistically significant (BAX 0.346±0.032 vs 0.302±0.037, Caspase-3 0.228±0.061 vs 0.158±0.065, BCL-2 0.090±0.047 vs 0.276±0.057, t value was 2.680, 2.361 and 7.575, respectively, all P<0.05). The expression levels of RUNX1 and apoptosis in pHNECs increased in a time-dependent manner after TNF-α exposure (P<0.05). Plasmid of sh-RUNX1-6 transfected silenced the expression of RUNX1 in pHNECs treated by TNF-α. After silencing RUNX1 in pHNECs apoptosis model, the protein levels of BAX and Caspase-3 were decreased, while the expression of BCL-2 was increased, the rate of apoptosis was decreased (P<0.05). Conclusions: RUNX1 is increased in NPs. Silencing RUNX1 can inhibit the apoptosis and reduce cell inflammatory damage of pHNECs induced by TNF-α.

[Utility of the type 3 portable monitor for the diagnosis of sleep disordered breathing in patients with stable heart failure].

Objective: To evaluate the utility of a type 3 portable monitor (PM) at home for the diagnosis of sleep disordered breathing (SDB) in patients with stable congestive heart failure (CHF). Methods: Seventy-six consecutive patients with CHF (61 males, 15 females, mean±standard deviation age (57.0±16.9) years) were enrolled from the sleep center of Peking university People's Hospital during January 2016 to January 2019, and underwent overnight, unattended home sleep apnea testing (HSAT) with a portable monitor followed by an overnight simultaneous polysomnogram (PSG) with in-laboratory portable monitor (in-lab PM) recording within one week. The consistency of apnea hypopnea index (AHI), obstructive sleep apnea index (OAI), central sleep apnea index (CAI) between HSAT and PSG as well as the in-lab PM and PSG were analyzed by Bland-Altman plot; the sensitivity and specificity of PM for the diagnosis of SDB in patients with CHF were evaluated. Results: The number of patients included in the final analysis were 65 in HSAT, 63 in in-lab PM and 65 in PSG. AHI [M(Q1,Q3)] was 26.1 (10.9,40.1) events/h by HSAT, 27.9 (11.3,43.2) events/h by in-lab PM, both were not different from AHI 29.0 (10.2,45.0) events/h by PSG (P>0.05). The AHI, OAI and CAI assessed by HSAT correlated significantly with those by PSG (r=0.892, 0.903, 0.831, P<0.05). Bland-Altman analysis of AHI, OAI, CAI by PSG versus HSAT showed a mean difference of 3.1 events/h, 0.8 events/h, 1.2 events/h; limits of consistency were -15.2 to 21.4 events/h, -9.7 to 11.3 events/h, -10.9 to 13.2 events/h, respectively. Based on a threshold of AHI ≥5 events/h, HSAT had 94.6% sensitivity, 75% specificity, compared to PSG. For detecting Cheyne-Stokes respiration (CSR), a sensitivity of 96.4%,a specificity of 97.2% were achieved, compared to PSG. Conclusion: Type 3 PM can be used to diagnose SDB in patients with CHF.

Differential expression of two C-type lectins in grass carp Ctenopharyngodon idella and their response to grass carp reovirus.

The cDNAs of two C-type lectins in grass carp Ctenopharyngodon idella, galactose-binding lectin (galbl) and mannose-binding lectin (mbl), were cloned and analysed in this study. Both of them exhibited the highest expression level in liver, whereas their expression pattern differed in early phase of embryonic development. Following exposure to grass carp reovirus (GCRV), the mRNA expression level of galbl and mbl was significantly up-regulated in liver and intestine.

Baicalin decreases SGK1 expression in the hippocampus and reverses depressive-like behaviors induced by corticosterone.

The present study was to investigate whether baicalin can prevent repeated exogenous corticosterone injection-induced depressive-like behaviors and explore its possible mechanisms. After a 21-day treatment with baicalin (10 and 20 mg/kg), sucrose preference in the sucrose preference test (SPT) and immobility time in forced swimming test (FST) were observed, serum corticosterone levels and brain-derived neurotrophic factor (BDNF) contents in the hippocampus were examined by enzyme-linked immunosorbent assay (ELISA). In addition, quantitative real-time polymerase chain reaction (qPCR) and western blot were used to detect the mRNA and protein expression in the hippocampus. The results showed that 21-day cortiscosterone injections caused depressive-like behaviors in mice, including the reduced sucrose preference and increased duration of immobility. Baicalin reversed these behavioral changes described above and restored serum corticosterone levels. Additionally, baicalin up-regulated the mRNA and protein expression of glucocorticoid receptor (GR) and BDNF, accompanied with the down-regulation of serum- and glucocorticoid-regulated kinase 1 (SGK1) in the hippocampus. Moreover, baicalin significantly increased the protein expression of 11ß-hydroxysteroid dehydrogenase-2 (11ß-HSD2) in the hippocampus. The present results confirmed the antidepressant-like effects of baicalin in a mice model of depression induced by corticosterone and suggested that its mechanism was possibly involved in reducing serum corticosterone and thereby increasing BDNF in the hippocampus.

Characterizations of four toll-like receptor 4s in grass carp Ctenopharyngodon idellus and their response to grass carp reovirus infection and lipopolysaccharide stimulation.

In this study, the subcellular localization, tissue distribution and response to grass carp reovirus (GCRV) infection and lipopolysaccharide (LPS) stimulation of four grass carp Ctenopharyngodon idellus toll-like receptor 4 (tlr4) genes were investigated. All four genes were constitutively expressed in all tissues studied, but the subcellular localization and tissue exhibiting the highest expression differed for each protein. Following GCRV infection, all the four tlr4s were upregulated in all tissues examined, and stimulation of C. idellus kidney (CIK) cells with LPS resulted in downregulation of all four tlr4s. These results provide a foundation for further investigation of tlr4 genes in bony fishes.

Enhancement of dissolution of cyclosporine A using solid dispersions with polyoxyethylene (40) stearate.

A solid dispersion containing cyclosporine A (CyA) and polyoxyethylene (40) stearate (PS) was prepared by the solvent-melt method and characterized by powder X-ray diffraction (PXRD), hot-stage microscopy (HSM), scanning electron microscopy (SEM) and dissolution studies. The crystalline peaks of CyA disappeared in the PXRD spectra of solid dispersions but were seen in those of physical mixtures, demonstrating the amorphous state of the drug in solid dispersions. The solubility of CyA in aqueous solutions of PS was increased linearly with increasing amount of PS in water. Dissolution of the drug from solid dispersions and physical mixtures was dramatically enhanced compared to the drug powder alone in water at 37 degrees C.

PEGylated recombinant human tumor necrosis factor alpha: preparation and anti-tumor potency.

AIM: To assess the merits of polyethylene glycol-modified recombinant human tumor necrosis factor alpha (PEG-rHuTNF-alpha). METHODS: The rHuTNF-alpha was modified with N-succinimidyl succinate monomethoxy polyethylene glycol (SS-PEG) of three different molecular weights. The PEG-rHuTNF-alpha was separated into fractions of various molecular weights by gel filtration chromatography. In vitro activities of various fractions were determined with L929 cell assay and in vivo anti-tumor potencies of main fractions were studied with respect to necrosis of S-180 solid tumor. RESULTS: The rHuTNF-alpha could be modified using SS-PEG under mild conditions. The main fraction of PEG5000-rHuTNF-alpha contained four PEG molecules, and PEG12000-rHuTNF-alpha and PEG20000-rHuTNF-alpha contained two PEG molecules, respectively. There was a higher activity when rHuTNF-alpha was coupled to less numbers of the same molecular weight PEG molecules. When PEG-rHuTNF-alpha was of the same molecular weight, rHuTNF-alpha modified with bigger molecular weight PEG molecules had a higher activity. PEG-rHuTNF-alpha was resistant to proteolysis, and over 70 % activity remained after 8 h, but the activity of rHuTNF-alpha was time-dependently diminished by incubation with bovine trypsin. PEG5000-rHuTNF-alpha (1500 IU per mouse) had a similar anti-tumor potency compared with rHuTNF-alpha (3000 IU per mouse). PEG12000-rHuT NF-alpha (1500 IU per mouse) had an increased anti-tumor potency compared with rHuTNF-alpha (3000 IU per mouse). In particular, PEG20000-rHuTNF-alpha at a dose of 1500 IU per mouse had a higher anti-tumor potency than rHuTNF-alpha at a dose of 6000 IU per mouse. CONCLUSION: PEG-modified rHuTNF-alpha could be more suitable for therapeutic use

PEGylated polycyanoacrylate nanoparticles as salvicine carriers: synthesis, preparation, and in vitro characterization.

AIM: To synthesized poly(methoxypolyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate) (PEGylated PHDCA) with polyethylene glycol (PEG, Mr = 5000), prepare PEGylated PHDCA and poly(n-hexadecyl cyanoacrylate) (PHDCA) nanoparticles loading salvicine and determine their in vitro characterizations. METHODS: The structure of PEGylated PHDCA was determined with 1H-NMR, 13C-NMR and Fourier transform infrared spectrum (FTIR). Its molecular weight was determined by gel permeation chromatography (GPC). Nanoparticles were prepared by emulsion/solvent evaporation method. RESULTS: 1H-NMR, 13C-NMR, and FTIR were consistent with structure of PEGylated PHDCA, whose average molecular weight is 6680. Entrapment efficiency could be determined by high pressure liquid chromatography (HPLC) method without endogenous interference at the retention time of salvicine. The entrapment efficiency was 92.6 % for PEGylated PHDCA nanoparticles and 98.9 % for PHDCA nanoparticles. The nanoparticles size was about 250 nm. The values of the zeta potential were obviously influenced by the composition of the copolymer. Compared with PHDCA nanoparticles (-23.1 mV), PEGylated PHDCA nanoparticles showed a low surface potential (-9.6 mV). Salvicine release from nanoparticles showed an initial burst effect, then a plateau for an extended period, and finally sustained release phase. CONCLUSION: These results showed that the PEGylated PHDCA nanoparticles could be an effective carrier for salvicine delivery in the respect of anti-tumor potency.

Stealth polycyanoacrylate nanoparticles as tumor necrosis factor-alpha carriers: pharmacokinetics and anti-tumor effects.

The objective of this study was to investigate the pharmacokinetics and in vivo anti-tumor effect of recombinant human tumor necrosis factor-alpha (rHuTNF-alpha) encapsulated in poly(methoxypolyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate) (PEG-PHDCA) nanoparticles. Our experimental results showed that PEG-PHDCA nanoparticles could extend the half-life of rHuTNF-alpha to 7.42 h and obviously change the protein biodistribution in tissues, and in particular, increase accumulation of rHuTNF-alpha in tumor. Compared with PHDCA nanoparticles and free rHuTNF-alpha, PEG-PHDCA nanoparticles loaded with rHuTNF-alpha showed higher antitumor potency at the same dose, which might be related to its higher accumulation in tumor tissues and longer plasma circulation time. Therefore, PEG-PHDCA nanoparticles could be an effective carrier for rHuTNF-alpha.

PEGylated recombinant human tumor necrosis factor alpha: pharmacokinetics and anti-tumor effects.

The aim of the present work was to investigate and assess the merit of PEGylated recombinant human tumor necrosis factor-alpha (rHuTNF-alpha) following our previous work. The rHuTNF-alpha was modified using activated polyethylene glycol (PEG), N-succinimidyl succinnate monomethoxy polyethylene glycol (SS-PEG). The pharmacokinetics and anti-tumor effect were investigated. The experimental results showed that PEGylated rHuTNF-alpha could obviously alter in vivo behavioral characteristics of rHuTNF-alpha. Among the synthesized PEG-rHuTNF-alphas with different PEG molecules, PEG20000-rHuTNF-alpha demonstrated the longest circulating half-life (24.8 h) which was about 50 times longer than that of rHuTNF-alpha (28.8 min). In addition, there was much more PEG20000-rHuTNF-alpha distributed into tumor tissues than other PEG-rHuTNF-alphas or rHuTNF-alpha with time, and PEG20000-rHuTNF-alpha also showed the highest anti-tumor potency. These results indicated that PEG20000-rHuTNF-alpha was a useful long circulating molecule with selective localization in tumor tissues and enhanced anti-tumor activity of rHuTNF-alpha.

PEGylated polycyanoacrylate nanoparticles as tumor necrosis factor-alpha carriers.

The aim of this study was to find an effective carrier for recombinant human tumor necrosis factor-alpha (rHuTNF-alpha). The influence of solvent systems containing poly(methoxy-polyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate) (PEGylated PHDCA) on the biological activity of rHuTNF-alpha was investigated. The PEGylated PHDCA nanoparticles loading rHuTNF-alpha were prepared with the double emulsion method. The influence of main experimental factors on the entrapment efficiency was evaluated by the Uniform Design. The physicochemical characteristics and in vitro release of rHuTNF-alpha from the nanoparticles were determined. The results showed that serum albumin such as human serum albumin (HSA) or bovine serum albumin (BSA) could play a protective action on rHuTNF-alpha in the preparation process. At > or =2.0% (w/v) HSA concentration, more than 85% of rHuTNF-alpha activity remained and the role of HSA was not affected by copolymer concentrations from 0.5 to 3.0% (w/v). The entrapment efficiency of the nanoparticles was about 60% and the nanoparticle size was about 150 nm. The nanoparticles were spherical in shape and uniform with the value of the zeta potential about -9 mV. The rHuTNF-alpha release from the nanoparticle showed an initial burst and then continued in a sustained fashion. The results showed that the PEGylated PHDCA nanoparticles could be an effective carrier for rHuTNF-alpha.

Pharmacokinetics of intragastric ipriflavone solid dispersion in rats.

AIM: To evaluate pharmacokinetic behavior of ipriflavone solid dispersion in rats. METHODS: The plasma concentrations of ipriflavone in rats were determined by HPLC with UV detector. RESULTS: Plasma concentration-time curves after ig ipriflavone solid dispersion 250 mg.kg-1 in rats were fitted with one-compartment model. Pharmacokinetic parameters were as follows: Ke = 0.21 h-1, T1/2Ke = 5.19 h, Ka = 1.71 h-1, T1/2Ka = 0.41 h, Tmax = 0.67 h, Cmax = 429 micrograms.L-1, AUC = 3916 micrograms.h.L-1; The relative bioavailability of ipriflavone solid dispersion was 323%. CONCLUSION: Ipriflavone in solid dispersion was absorbed more effectively than that in physical mixture in rats.

Preparation and dissolution property of ipriflavone solid dispersion.

AIM: To prepare and identify ipriflavone (IP) solid dispersion, and determine its dissolution property. METHODS: The solvent method was used for preparation and differential scanning calorimetry (DSC), X-ray diffraction and infrared spectrophotometry for identification of IP solid dispersion. The dissolution of the dispersion was determined with paddle method. RESULTS: The dissolution of IP solid dispersion consisting of IP and povidone-k30 (PVP) (1:8) in artificial gastric juice is 6.15 times as high as that of IP alone. The DSC curves, X-ray diffraction patterns and infrared spectrophotometries of IP have been changed obviously by the dispersion. CONCLUSION: The dissolution of IP is increased by solid dispersion method.

An improved HPLC assay for ciprofloxacin in biological samples.

A simple and sensitive high performance liquid chromatographic method was developed for determination of ciprofloxacin (Cip) in 0.1 ml of rabbit and human serum and CSF. Separation of Cip and pipemidic acid (internal standard) was achieved on Nucleosil C18 (4.6 mm x 250 mm) using fluorescence detection with lambda exc 274 nm and lambda emi 418 nm. The mobile phase was composed of acetonitrile and phosphate buffer 10 mmol.L-1 (pH 2.7) containing tetrabutylammonium hydrogen sulfate 5 mmol.L-1 (18:82, vol:vol) and the flow rate was set at 1.0 ml.min-1. The retention times were 5.9 min for Cip and 4.0 min for pipemidic acid. The inter-day coefficient of variation was < 6.97% (n = 5) and the intra-day coefficient of variation was < 3.33% (n = 5). The limits of detection were 3 ng.ml-1 serum and 5 ng.ml-1 CSF, (r > or = 0.9996). Application of this method was demonstrated with simultaneous measurements of concentration-time profiles of Cip in rabbit serum and CSF during iv infusions at constant rates of 0.33, 1.0, and 2.5 mg.kg-1.h-1.

Clinical study on the pharmacokinetics of lithium carbonate.

To investigate clinical pharmacokinetics of lithium carbonate, 28 hospitalized mental patients and 8 healthy volunteers were studied. The serum concentration/time profile in the single dose peroral experiments could be fitted to a pharmacokinetic model using an open two compartmental system. Three methods, 72-h and 24-h residual method and Ritschel's repeated one-point method were used and the results compared with one another. According to the results of this paper, we suggest the use of Ritschel's repeated one-point method for clinical dosage regimen determination.

Effects of phenobarbital steady state levels on antipyrine clearance and distribution in the rat.

A group of 15 rats received two intravenous bolus doses of antipyrine (15 mg/kg) separated by a 57 hour infusion (with bolus dose) of phenobarbital. Phenobarbital bolus doses and infusion rates were based on a preliminary pharmacokinetic study (7 rats) and were varied to achieve a broad range of steady state levels. Antipyrine and phenobarbital blood levels were measured by high pressure liquid chromatography. Antipyrine kinetics obeyed first order monoexponential decay, and the parameters (clearance, volume, half-life) were determined. Antipyrine clearance increased in all animals during phenobarbital infusions with a per cent increase ranging between 54.6 and 269 per cent. However, no significant correlation was found between the per cent increase in antipyrine clearance and phenobarbital concentration (r = 0.19). The volume of distribution of antipyrine increased in 14 of 15 animals with increases ranging between 7.7 and 45.8 per cent.