Assessment of quality deviation of pork and salmon due to temperature fluctuations during superchilling.

Superchilling is an emerging technology for meat preservation; however, the temperature changes during the process have been commonly ignored. Thus, the effects of temperature fluctuations on meat quality during superchilling are yet to be evaluated. In our study, pork loins and salmon fillets were stored for several days (0, 8, 15, 23, and 30 d) under different temperature fluctuations based on -3.5 ℃ as the target temperature. The results showed that after 15 d of superchilling storage, the values of total volatile basic nitrogen, total viable count, and lipid oxidation were significantly (P<0.05) altered in the ±2.0 ℃ fluctuation group compared with the constant temperature group. On the contrary, there was no significant difference in these parameters between the ±1.0 ℃ fluctuation group and the constant temperature group after 30 d of storage. In addition, irregular temperature changes significantly accelerated the modulation of various indicators. In brief, temperature fluctuations and irregular temperature changes accelerated the destruction of muscle structural integrity, increased the water loss, gradually widened the water loss channels, and thereby reduced the edibility by accelerating the spoilage of meat.

Glycopolymer-Cell-Penetrating Peptide (CPP) Conjugates for Efficient Epidermal Growth Factor Receptor (EGFR) Silencing.

Overexpression of epidermal growth factor receptor (EGFR) is observed in multiple cancers such as colorectal, lung, and cervical solid tumors. Regulating the EGFR expression is an efficient strategy to manage these malignancies, and it can be achieved by using short interfering RNA (siRNA). Cell-penetrating peptides (CPPs) demonstrated an excellent capability to enhance the cellular uptake of siRNA, but high knockdown efficiencies have not been achieved due to endosomal entrapment. In this work, Schiff's base reaction was used to modify a block {P[LAEMA(2-lactobionamidoethyl methacrylamide)37]-b-P[FPMA(4-formyl phenyl methacrylate)2-st-DMA(N,N-dimethylacrylamide)2], P2} and two statistical [P(LAEMA23-st-FPMA3) (P3) and P(LAEMA25-st-FPMA2-st-DMA2) (P4)] aldehyde-based and galactose-based polymers, prepared via reversible addition-fragmentation chain-transfer (RAFT) polymerization. An arginine-rich peptide (ARP, KRRKRRRRRK) was used as a cell-penetrating peptide (CPP) and conjugated to the polymers via a Schiff base reaction. The resulting glycopolymer-peptide conjugates were utilized to condense the siRNA to prepare polyplexes with multivalent CPPs (MCPPs, a nanoparticle with multiple copies of the CPP) to enhance the endosomal escape. The polyplexes have different surface properties as determined by the architecture of polymers and the insertion of dimethyl amide moieties. The enhancement of cellular internalization of ARP was observed by labeling the polyplexes with fluorescein isothiocyanate (FITC)-siRNA showing a localization of polyplexes in the cytoplasm of a HeLa (cervical cancer) cell line. In the in vitro EFGR silencing study, the statistical glycopolymer-peptide (P3-P) polyplexes had superior EGFR silencing efficiency in comparison with the other polymers that were studied. Furthermore, P3-P polyplexes led to less off-targeting silencing than lipofectamine 3000. These encouraging results confirmed the potency of decorating galactose-based polymers with CPP, like ARP for their application in siRNA delivery and management of cervical carcinomas.

Synthesis and characterization of a PAMAM dendrimer nanocarrier functionalized by HA for targeted gene delivery systems and evaluation in vitro.

Poly(amido-amine) (PAMAM), one of the most widely studied dendrimers in the field of drug and gene delivery, can enhance the stability of DNA and deliver it to cell cytosol; hyaluronic acid (HA), a simple disaccharide unit, can polymerize and is considered a polymer of non-immunogenicity, which has an intrinsic targeting property for many cancer cells by interacting with CD 44. In this study, we had synthesized and characterized a series of PAMAM modified by HA. and PAMAM was conjugated by HA with different grafting density (5%, 15%, 25%) and molecular weight (HA3850, HA17200). We had investigated the particle size, zeta potential and Agarose gel electrophoresis assays of polyplexes. Besides, the cytotoxicity, transfection efficiency and the mechanisms of transfection of new polyplexes were assessed following in vitro transfection in Hela, Bel-7402 and HepG2 cells lines. In the results, modified by HA, the cytotoxicity of polymer had reduced and the size of some polymers also below 200 nm in appropriate weight ratio, and transfection efficiency had also close to the polyplexes G4 PAMAM/DNA were observed. Compared with the unmodified dendrimers compounds, the DNA delivering capacity of PAMAM G4-HA3850-5% and PAMAM G5-HA3850-5% had improved in cancer cells line. It is a potential candidate used for targeted gene delivery.

Survivals of the Intraoperative Motor-evoked Potentials Response in Pediatric Patients Undergoing Spinal Deformity Correction Surgery: What Are the Neurologic Outcomes of Surgery?

STUDY DESIGN: This is a retrospective cases study from a prospective patient register. OBJECTIVE: To clarify the clinical implication regard to the survivals of motor-evoked potential (MEP) response. SUMMARY OF BACKGROUND DATA: Intraoperative neurophysiological monitoring has become an essential component for decreasing the incidence of neurological deficits during spine surgeries. Significant motor-evoked potential (MEP) loss but does not vanish completely is common especially in some high-risk and complicated pediatric spine deformity surgeries. METHODS: A total of 1820 young patients (mean age = 12.2 years) underwent spinal deformity correction were mainly analyzed. Intraoperative monitoring (somatosensory-evoked potential, MEP, free-run electromyography, free-run electromyography) and postoperative neurologic outcomes were mainly analyzed in this study. All patients with monitoring alerts were divided into two groups: group 1, intraoperative MEP recovery group; and group 2, no obvious MEP recovery group. Moreover, the patients would be followed up strictly if he/she showed IOM alerting. The surviving MEP response was identified as significant monitoring alerts (80%-95% MEP Amp. loss) associated with high-risk surgical maneuvers. RESULTS: The results showed that there were 32 pediatric patients (group 1, 21 cases and group 2, 11 cases) presenting significant MEP monitoring alerts (80%-95% loss) relative to baseline. The patients in group 1 presented the partial/entire signal recovery from MEP alerts and they did not show spinal cord deficits postoperation. The patients in group 2 without obvious intraoperative MEP recovery showed different levels of new spinal deficits, no patient showed postoperative complete paraplegia or permanent spinal cord/nerve root deficits. CONCLUSION: When the intraoperative MEP changes significant and persistent but without totally disappeared, the rate of postoperative neural complication is relatively low. The chance of recovery of these neurological deficits is very high. Therefore, this phenomenon may be used to predictive of nonpermanent paraplegia. LEVEL OF EVIDENCE: 3.

Preparation of gene drug delivery systems of cationic peptide lipid with 0G-PAMAM as hydrophilic end and its biological properties evaluation.

As an efficient gene delivery, non-viral vectors should have high transfection efficiency, excellent endosomal escape, low cytotoxicity, and the ability to rapidly release the gene into the cytoplasm.Cationic liposome have been widely used as efficient gene carriers, but the cytotoxicity, rapid degradation and low cellular uptake are major drawback impeding its further appolication. Herein, with double lauric acid as hydrophobic chains, tartaric acid as skeleton, 0 generation PAMAM modified with lysine as hydrophilic head, a new type cationic peptide lipid was synthetised. The alkyl chain promote lipid across cell membranes and with membrane fusion, 0 generation PAMAM modified with lysine hydrophilic end amino can contain a large number of protons which can change into ammonium and combine with the DNA negatively charge phosphate groups. It is expected that this carrier has low toxicity, high transfection efficiency and targeting property. By adjusting the cationic liposome/gene weight ratio, the transfection system was optimized to improved gene transfection efficiency, reduce cytotoxicity, and increase property and stability, etc.

Multi-armed poly(L-glutamic acid)-graft-polypropyleneinime as effective and serum resistant gene delivery vectors.

A new series of multi-armed MP-g-PPI dendrimers were synthesized by polymerization of BLG-NCA using G2.0PPI as macromolecular initiator and subsequent aminolysis with G1.0PPI or G2.0PPI. The chemical structure and composition of the MP-g-PPI dendrimers were characterized by Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance spectroscopy ((1)H NMR). The MP-g-PPI showed a great ability to combine with pDNA to form complexes, which protect the pDNA from nuclease degradation. Dynamic light scattering (DLS) measurement illustrated that the sizes of complexes were in range of 111-219 nm. The transmission electron microscope (TEM) and atomic force microscope (AFM) observation showed that the morphology of these complexes was spherical. The MTT assay demonstrated that cytotoxicity of the MP-g-PPI was lower than that of PEI 25K. The in vitro transfection test indicated that MP-g-PPI gene vectors displayed relative high transfection efficiency than that of PEI 25K and Lipofectamine 2000 in serum-containing medium. Furthermore, MP-g-PPI at the weight ratio of 7.5 displayed better serum-resistant capability than that of PEI 25K and Lipofectamine 2000. The above facts revealed that multi-armed MP-g-PPI dendrimers may be promising gene vectors with low cytotoxicity, high transfection efficiency and serum-resistant ability.

Study on galactose-poly(ethylene glycol)-poly(L-lysine) as novel gene vector for targeting hepatocytes in vitro.

A non-viral gene-delivery system has been used to deliver plasmid DNA into specific cell types because of its safety and ease of manufacture. Receptor-mediated gene transfer is currently a promising gene-delivery technique. To specifically target genes to asialoglycoprotein receptor of hepatocytes, a galactose moiety was combined into the poly(ethylene glycol) (PEG)-terminal end by reductive coupling using lactose and sodium cyanoborohydride. A synthesis method of conjugating poly(L-lysine) (PLL) derivatives with terminally galactose-graft-PEG was developed using ring-opening polymerization of N(ε)-benzyloxycarbonyl-L-lysine-N(α)-carboxyan-hydride (Z-Lys-NCA) initiated onto galactose graft amine-terminated PEG (galactose-PEG-NH2) as a macro-initiator. The synthesis was characterized with ¹H-, ¹³C-NMR, IR and UV spectroscopy, and all of them successfully verified the formation of the co-polymers. The gel-retardation assay of the complexes between galactose-PEG-PLL and plasmid DNA indicated that these polymeric gene carriers demonstrated the potent ability to condense plasmid DNA electrostatically as well as PLL. The particle size and zeta potential of polymer/DNA complexes were measured, and their cytotoxicity and transfection efficiency in different cells were evaluated. The results indicate that galactose-PEG-PLL can form a complex with plasmid DNA and serve as an effective gene-delivery carrier with lower cytotoxicity compared to that of PLL. Transfection experiments clearly showed that galactose-PEG-PLL effectively delivered DNA into hepatoma cells in vitro. Such data demonstrates that galactose and its complex with plasmid DNA may serve as a safe and effective gene-transfer system targeting hepatocytes.

[Survey on host animal and molecular epidemiology of hantavirus in Chuxiong prefecture, Yunnan province].

OBJECTIVE: To determine the hosts of hantavirus (HV) and its molecular epidemiological characteristics, to provide evidence for prevention and control on hemorrhagic fever with renal syndrome (HFRS). METHODS: Rodents were captured by a special trap within the residential area. The antigens of HV in lung tissues were detected by direct immuno-fluorescence assay (DFA). Nucleotide sequences of HV were amplified by RT-PCR with HV genotype-specific primer. The amplified genes were then sequenced. Phylogenetic tree were built on nucleotide sequence with ClustalX 1.83 software. RESULTS: 1421 rodents were captured and classified into 8 species of 4 Genera in the epidemic area within 10 counties of Chuxiong prefecture, Yunnan province, between 2005 and 2006. Out of the 1421 rodents, 1056 (74.31% ) of them were Rattus norvegicus and 280 (19.70%) belonged to Rattus flavipectus. The antigens of HV were detected by DFA in lung tissues and the total positive rate of HV was 5.15% (53/ 1029). After applying the sequencing nucleotide method to the 53 positive specimens, data showed that 21 specimens were positive and all of them belonged to Seoul type (15 samples were from Rattus norvegicus, 4 samples Rattus flavipectus, 2 samples Rattus nitidus). The partial S segments from 12 specimens were sequenced which appeared homologic with R22, L99 and HLD65 from GenBank in relatively high level (87.1% -99.7%). When compared to 76-118 strain of Hantaan type, their homologic degree was only 64.4%-69.1%. Results from Phylogenetic analysis showed that 12 specimens belonged to Seoul type. As for their homology, they were significantly similar to Seoul type and could be tentatively divided into two subtypes S1 and S3. CONCLUSION: It was confirmed that the Seoul type virus, as HFRS's pathogenetic agent mainly carried by rats, prevailed widely in Chuxiong prefecture. Owing to the local ecological environment, we also noticed the characteristics of different HV subtypes among Seoul type.

Poly(ethylene glycol)-block-polyethylenimine copolymers as carriers for gene delivery: effects of PEG molecular weight and PEGylation degree.

An ideal gene carrier is required both in safety and efficiency for transfection. Polyethylenimine (PEI), a well-studied cationic polymer, has been proved with high transfection efficiency, but is reported as toxicity in many cell lines. In this study, PEI was coupled with polyethylene glycol (PEG) to reduce its cytotoxicity. PEG-PEI copolymers were synthesized with isoporon diisocyanate (IPDI) in two steps. A set of PEG-PEI with different PEG molecular weights (MWs) and amounts of PEG were synthesized. The molecular structure of the resulting copolymers was evaluated by nuclear magnetic resonance spectroscopy ((1)H NMR), infrared spectroscopy (IR), and gel permeation chromatography (GPC), all of which had successfully verified formation of the copolymers. The particle size and zeta potential of polymer/DNA complexes were measured, and their cytotoxicity and transfection efficiency in Hela cells were evaluated. We found that the copolymer block structure significantly influenced not only the physicochemical properties of complexes, but also their cytotoxicity and transfection efficiency. PEG (5 kDa) significantly reduced the diameter of the spherical complexes. The zeta potential of complexes was reduced with increasing amount of PEG grafting. Cytotoxicity was dependent not on PEG MW but on the amount of PEG grafting. Copolymer PEG-PEI (2-25-1) with 1.89 PEG (2 kDa) was proved to be more efficient for in vitro gene transfer. In conclusion, PEG MW and the degree of PEGylation were found to significantly influence the biological activity of PEG-PEI/DNA complexes. These results provide new sights into the studies using block copolymer as gene delivery systems.