Complete genome sequencing and analysis of six enterovirus 71 strains with different clinical phenotypes.

BACKGROUND: Hand, foot and mouth diseases (HFMD) caused by enterovirus 71(EV71) presents a broad spectrum of clinical manifestations ranging from mild febrile disease to fatal neurolocal disease. However, the mechanism of virulence is unknown. METHODS: We isolated 6 strains of EV71 from HFMD patients with or without neurological symptoms, and sequenced the whole genomes of the viruses to reveal the virulence factors of EV71. RESULTS: Phylogenetic tree based on VP1 region showed that all six strains clustered into C4a of C4 sub-genotype. In the complete polypeptide, 298 positions were found to be variable in all strains, and three of these positions (Val(P814)/Ile(P814) in VP1, Val(P1148)/Ile(P1148) in 3A and Ala(P1728)/Cys)/Val(P1728) in 3C) were conserved among the strains with neurovirulence, but variable in strains without neurovirulence. In the 5'-UTR region, it showed that the first 10 nucleotides were mostly conserved, however from the 11th nucleotide, nucleotide insertions and deletions were quite common. The secondary structure prediction of 5'-UTR sequences showed that two of three strains without neurovirulence (SDLY11 and SDLY48) were almost the same, and all strains with neurovirulence (SDLY96, SDLY107 and SDLY153) were different from each other. SDLY107 (a fatal strain) was found different from other strains on four positions (C(P241)/T(P241), A(P571)/T(P571), C(P579)/T(P579) in 5'-UTR and T(P7335)/C(P7335) in 3'-UTR). CONCLUSIONS: The three positions (Val(P814)/Ile(P814) in VP1, Val(P1148)/Ile(P1148) in 3A and Ala(P1728)/Cys(P1728)/Val(P1728) in 3C), were different between two phenotypes. These suggested that the three positions might be potential virulent positions. And the three varied positions were also found to be conserved in strains with neurovirulence, and variable in strains without neurovirulence. These might reveal that the conservation of two of the three positions or the three together were specific for the strains with neurovirulence. Varation of secondary structure of 5'-UTR, might be correlated to the changes of viral virulence. SDLY107 (a fatal strain) was found different from other strains on four positions, these positions might be related with death.

Ultrasensitive and green electrochemical immunosensor for mycotoxin ochratoxin A based on phage displayed mimotope peptide.

Here, we demonstrated a new approach for development of an ultrasensitive and green electrochemical immunosensor for Ochratoxin A (OTA). Phage displayed mimotope peptide of OTA was used as mimics of conventional competing antigen, which is chemical synthesized with toxic mycotoxins OTA as raw material, in a competitive sensing platform. The working electrode was modified by polyethylene glycol (PEG) for the purpose of immobilizing antibody effectively. Under the optimized test condition, the limit of detection (LOD) of the established immunosensor was 2.04 fg/mL, and the linear range was 7.17-548.76 fg/mL. Specific measurement of this established method was conducted by testing cross-reactivity of other common mycotoxins, the result showed that mimotope peptide-based immunosensor has negligible cross-reactivity with other mycotoxins. Furthermore, the novel concept of phage displayed mimotope peptide-based immunosensor may provide a potential application in general method for the ultrasensitive detection of various toxic small molecules in food.

Identification of a functional polymorphism within the 3'-untranslated region of denticleless E3 ubiquitin protein ligase homolog associated with survival in acral melanoma.

BACKGROUND: High expression of denticleless E3 ubiquitin protein ligase homologue (DTL) correlates with poor disease-free survival and overall survival in cutaneous melanoma, but the molecular features and clinical significance of this gene in acral melanoma (AM) remain unclear. METHODS: The expression levels of DTL were compared between AM and benign melanocytic nevi using existing Gene Expression Omnibus data and validated in fresh frozen tissues. Two candidate tag single-nucleotide polymorphisms (SNPs) in the 3'-untranslated region (3'UTR) of DTL in patients with AM were sequenced and analysed for their association with survival in a discovery cohort (n = 570), and the significant SNP was subjected to a replication cohort (n = 201). The expression of DTL was evaluated by immunohistochemistry. The microRNA interacting with rs11275300:C > G was predicted using in silico target prediction tools and validated by in vitro analysis. RESULTS: DTL was overexpressed in AM compared with benign melanocytic nevi. rs11275300:C > G was found to be significantly associated with progression-free survival and overall survival of patients with AM in both cohorts and the combined cohort. Furthermore, the DTL expression level in the patients with the rs11275300:G allele was higher than that in patients with the CC genotype. In vitro analysis demonstrated that DTL was a direct target of hsa-miR-4672, and the rs11275300:G allele interfered with the binding affinity of hsa-miR-4672 with the 3'UTR of DTL and thereby increased DTL expression. CONCLUSION: The rs11275300:G allele in the 3'UTR of DTL may lead to a poor prognosis and allele-specific increase in the expression of DTL by post-transcriptional regulation in AM.

[Advances in the study of excipient inhibitors of intestinal P-glycoprotein].

P-glycoprotein (P-gp) located in the apicalmembranes of intestinal absorptive cells is an energy-dependent efflux pump which can reduce the bioavailability of a wide range of substrate drugs. There is increasingly interest in enhancing the bioavailability of these molecules by inhibiting intestinal P-gp. A classification of excipient inhibitors of intestinal P-gp nonionic surfactants, poly (ethylene glycol), derivates of beta-cyclodextrin and thiolated chitosan will be presented and then the inhibition mechanism will be discussed. Compared with traditional P-gp inhibitor, excipient inhibitors appear to have minimal nonspecific pharmacological activity, thus potential side effects can be mostly avoided. These excipient inhibitors, which hold the promise of replacing the traditional ones, will be extensively employed to significantly improve the intestinal absorption of poorly soluble and absorbed drugs as a result of P-gp inhibition, and thus to enhance the bioavailability of these drugs. However, the further studies of both the mechanism and clinical application are urgently needed.

[Effect of polyoxyl ether analogous surfactants on the activity of cytochromes P450 3A in rats in vivo].

To evaluate the effects of p-octyl polyethylene glycol phenyl ether (Triton X-100), polyoxyl 35 caster oil (EL35) and polyoxyl 40 hydrogenated caster oil (RH40) on the activity of Cytochrome P450 3A (CYP3 As) in vivo. Rats were administered with saline, ketoconazole (75 mg x kg(-1) x d(-1)), Triton X-100 (30 mg x kg(-1) x d(-1)), EL35 (150 mg x kg(-1) x d(-1)) and RH40 (150 mg x kg(-1) x d(-1)) intragastrically for 5 consecutive days, and then given midazolam 10 mg x kg(-1) 20 min after the last treatment of ketoconazole or three surfactants with the same dose through duodenal administration. Pharmacokinetics parameters for midazolam and its metabolite 1'-hydroxymidazolam were estimated from the plasma concentration-time data by a noncompartmental approach. The results showed that multiple dose administration of Triton X-100, EL35 and RH40 decreased the ratios of 1'-hydroxymidazolam and midazolam AUC0-infinity from 1.14 to 0.90, 1.03 and 0.64, respectively. In contrast, multiple dose administration of ketoconazole caused the ratios of 1'-hydroxymidazolam and midazolam a significant decrease to 0.50. This study indicated that Triton X-100 and EL35 would have no inhibition on CYP3A, while RH40 had significant inhibition on CYP3A. Therefore, RH40 might be used to prepare drug formulations in pharmaceutical industry and would increase the bioavailability of some drugs transformed by CYP3As and further lead to significant clinical pharmacologic effects.

Self-organized polymer aggregates with a biomimetic hierarchical structure and its superhydrophobic effect.

Nature always gives us inspirations to fabricate functional materials by mimicking the structure design of biomaterials. In this article, we report that polymeric aggregates with morphology similar to the papilla on lotus leaf can be self-organized in the polymer solution by adding 16 wt% water into 5 mg/ml polycarbonate solution in N,N'-dimethylformamide. The hierarchically structured aggregates at micro- and nano-scale alone show superhydrophobic effect without the need of modification with low surface energy compound. Small amount of liquid can be wrapped by the aggregates to form the so-called liquid marble. Influence of the amount of water added into the solution on the morphology of resultant polymer aggregates was investigated. By using the hierarchical aggregates as the surface building blocks, superhydrophobic coating with a static water contact angle larger than 160 degrees and sliding angle less than 5 degrees (for a water drop of 5 microl) was formed. Other solutions, like acid, basic and blood plasma are also repelled on the coating.

[The inhibitory effect of pluronic on P-glycoprotein drug pump].

To investigate the inhibitory effect of Pluronic on P-glycoprotein (P-gp) drug efflux pump, Caco-2 cells and animal models were established to study the influence of Pluronic on celiprolol transport across Caco-2 cell monolayer and intestinal mucous membrane with verapamil set as a positive control. Drug concentration was measured by HPLC and the apparent permeability coefficient (P(app)), absorption rate constant (k(a)) and the effective permeability coefficient (P(eff)) were calculated. P(app) of basolateral to apical side and apical to basolateral side was (2.10 +/- 0.13) x 10(-6) and (0.333 +/- 0.018) x 10(-6) cm x s(-1), respectively. Transports of celiprolol across Caco-2 cell monolayer were influenced by both verapamil and Pluronic. The absorption constants (k(a)) of celiprolol at duodenum, jejunum, ileum, and colon were (0.09 +/- 0.03), (0.14 +/- 0.04), (0.11 +/- 0.03) and (0.05 +/- 0.02) h(-1), k(a) of celiprolol in verapamil group were (0.14 +/- 0.03), (0.24 +/- 0.02), (0.25 +/- 0.03) and (0.23 +/- 0.02) h(-1), and k(a) of celiprolol in Pluronic group were (0.13 +/- 0.02), (0.22 +/- 0.02), (0.22 +/- 0.03) and (0.20 +/- 0.03) h(-1), respectively. Pluronic showed significant effect on inhibiting P-gp of Caco-2 cell and intestinal mucosa in rats.

[Relationship between beta amyloid protein 1-40 and post-operative delirium after oral and maxillofacial surgery in elderly patients].

OBJECTIVE: To determine the incidence of post-operative delirium after oral and maxillofacial surgery under general anesthesia in elderly patients, and to examine its association with plasma concentrations of beta amyloid protein 1-40 (Abeta1-40). METHODS: Fifty patients underwent elective oral and maxillofacial surgery were divided into two groups: Group C (n=20) aged from 20 to 60 years old, and Group T (n=30) aged from 62 to 78 years old. The two group received the same general anesthesia. Delirium rating scale-revised-98 (DRS-R-98) was used as an instrument to diagnose and access the postoperative delirium of the two groups. The scores of DRS-R-98 were recorded before operation (T0) and at 24 h (T1), 48 h (T2), 72 h(T3) and 96 h(T4) after the operation. Serial measurements of serum concentrations of Abeta1-40 were also performed at the same time. RESULTS: The incidence of post-operative delirium after oral and maxillofacial surgery in Group T was 20.0%. The concentrations of plasma Abeta1-40 in group T were much higher than group C at TO, T1, T2 and T3 significantly (P < 0.01). The concentrations of plasma Abeta1-40 at T1 and T2 were higher than those at TO in the same group (P < 0.05). The scores of DRS-R-98 in Group T at T3 and T4 were much higher than those at T1 and Group C significantly (P < 0.01). CONCLUSION: The constant increase of plasma Abeta1-40 may be one of the important factors related to post-operative delirium in elderly patients underwent oral and maxillofacial surgery.