Transcriptomic analysis of Andrias davidianus meat and experimental validation for exploring its bioactive components as functional foods

Andrias davidianus(Chinese giant salamander,CGS)is the largest and oldest extant amphibian species in the world and is a source of prospective functional food in China.However,the progress of functional peptides mining was slow due to lack of reference genome and protein sequence data.In this study,we illustrated full-length transcriptome sequencing to interpret the proteome of CGS meat and obtain 10703 coding DNA sequences.By functional annotation and amino acid composition analysis,we have discovered various genes related to signal transduction,and 16 genes related to longevity.We have also found vast variety of functional peptides through protein coding sequence(CDS)analysis by comparing the data obtained with the functional peptide database.Val-Pro-Ile predicted by the CDS analysis was released from the CGS meat through enzymatic hydrolysis,suggesting that our approach is reliable.This study suggested that transcriptomic analysis can be used as a reference to guide polypeptide mining in CGS meat,thereby providing a powerful mining strategy for the bioresources with unknown genomic and proteomic sequences.

Site-specific determination of TTR-related functional peptides by using scanning tunneling microscopy

For the design and optimization of functional peptides, unravelling the structures of individual building blocks as well as the properties of the ensemble is paramount. TI＇R1, derived from human transthyretin, is a fibril-forming peptide implicated in diseases such as familial amyloid polyneuropathy and senile systemic amyloidosis. The functional peptide TTR1-RGD, based on a TFR1 scaffold, was designed to specifically interact with cells. Here, we used scanning tunneling microscopy （STM） to analyze the assembly structures of TTRl-related peptides with both the reverse sequence and the modified forward sequence. The site- specific analyses show the following： i） The TIR1 peptide is involved in assembly, nearly covering the entire length within the ordered [3-sheet structures, ii） For TTR1-RGD peptide assemblies, the TTR1 motif forms the ordered [3-sheet while the RGDS motif adopts a flexible conformation allowing it to promote cell adhesion. The key site is clearly identified as the linker residue Gly13. iii） Close inspection of the forward and reverse peptide assemblies show that in spite of the difference in chemistry, they display similar assembling characteristics, illustrating the robust nature of these peptides, iv） Glycine linker residues are included in the ^-strands, which strongly suggests that the sequence could be optimized by adding more linker residues. These garnered insights into the assembled structures of these peptides help unravel the mechanism driving peptide assemblies and instruct the rational design and optimization of sequence- programmed peptide architectures.

The Level of Serum Intact Terminal Peptide of Procollagen and Its Clinical Significance in Patients with Chronic Keshan Disease

Objectives To evaluate the changes of serum intact terminal peptide of procollagen in patients with chronic Keshan disease (KD) and investigate their clinical significance. Methods The concentrations of serum intact N-terminal peptide of type procollagen (P NP) and intact N-terminal peptide of type procollagen were measured by radioimmunoassay in 35 patients with chronic KD and 31 normal control. Doppler ultrasounds was used to determine several parameters of left ventricular systole and diastole functions. Results The concentration of serum P NP (74.07±16.74)μg/L and the ratio of P NP/ P NP (18.02 ±4.60) in chronic KD were significantly increased as compared to the control (39.63±12.07 μg/L, 12.12±4.24; P< 0.001). Serum P NP (4.19±0.64)μg/L in chronic KD was higher than that in the control (3.36±0.65 μg/L,P < 0.001) too. The higher of serum concentration of P NP and the ratio of P NP/ P NP, the worse of cardiac function in patients with chronic KD. A negative correlation was found between serum P NP/ P NP, P NP and VE/VA, LVEF (γ=-0.4502, -0.4608, P< 0.01 and γ=-0.3936, -0.3904, respectively; P<0.05). Conclusions These findings suggested that tissue synthesis of collagen type and type was abnormally increased in chronic KD. On the other hand, our results indicated that P NP and P NP were related to several functional alterations of the left ventricle. Serum procollagen peptide measurements might therefore provide indirectly diagnostic information on myocardial fibrosis associated with chronic KD.

Multifunctional building elements for the construction of peptide drug conjugates

Engineering of smart building molecules is key basis in designing intelligent drug delivery systems.As an emerging sophisticated delivery system strategy,the powerful functions of peptide drug conjugates(PDCs)are attributed to a smart linker and multifunctional peptide domain.Peptides exhibit a wide range of functions and properties,including easy chemical synthesis and versatile modification,tunable biocompatibility,diversified self-assembled nanostructures,specific recognition/binding,and deep penetration of the cell membrane/extracellular matrix.In addition,various types of linkers enable PDCs to release drugs responsively according to the disease microen-vironment or treatment needs.Owing to these inherent advantages,PDCs have been widely explored for drug delivery.Herein,the latest developments in functional peptides and linkers commonly used to construct smart PDCs are reviewed.The purpose is to bring widespread attention to PDC design strategies and their contribution to fighting various diseases,as well as to provide guidance for research on intelligent PDC drug delivery systems.

Correlations among Persistent Viral Infection,Heart Function and Chinese Medicine Syndromes in Dilated Cardiomyopathy Patients

Objective：To investigate the correlations among persistent viral infection,heart function and Chinese medicine（CM） difined-syndromes in patients with dilated cardiomyopathy（DCM）.Methods：Fifty patients with DCM in the First Affiliated Hospital of Zhejiang Chinese Medical University from October 2009 to December 2011 were selected as the research subjects,and 30 healthy people were simultaneously selected as the normal control group to detect persistent viral infections after admission.The CM syndrome type and grade of heart function were then evaluated.The expression level of Coxsackie adenovirus receptor（CAR） was detected using the flow cytometry（FCM） technique,coxsackie virus RNA（CVB-RNA） using reverse transcription polymerase chain reaction（RTPCR）,and the plasma brain natriuretic peptide（BNP） level with a Triage meter plus diagnosis instrument.Finally,the parameters such as left ventricular end diastolic diameter（LVEDd） and left ventricular ejection fraction（LVEF） were measured by ultrasonic cardiogram.Person correlation analysis was used for measured data,Spearman correlation analysis for rating data,and the Chi-square test for numerical data.Results：CVB-RNA was positive in 22 patients（44%） with DCM,while only 6 cases（20%） were CVB-RNA-positive in the normal control group,with a significant difference between the two groups（P〈0.01）.The expression level of CAR was significantly elevated in the DCM group compared with the normal control group（P〈0.01）.In CVB-RNA-positive patients（22 cases）,the expression level of CAR was significantly higher than in CVB-RNA-negative patients（28 cases;P〈0.01）.In the DCM patients,there was a positive correlation between the CAR expression and the BNP level（r=0.34,P〈0.05）,while no significant difference was found between the CAR expression and the LVEF and LVEDd（r=-0.32,0.30,P〉0.05）.There was no clear correlation between virus infection and the CM syndrome types in DCM patients（r=-0.22,P〉0.05）.According to the sequence of syndrome types：phlegm → qi deficiency → blood stasis → hydroretention with asthenic yang（from low to high）,a positive correlation was existed between the BNP levels and CM syndrome types（r=0.139,P〈0.05）.Conclusion：The expression of CAR on the surface of white cells could be used to detect persistent viral infection.The expression level of CAR and heart function in DCM patients were highly correlated.The expression level of BNP may serve as an objective index for differentiating CM syndromes for patients with DCM.

Influence of Cardiac Structure, Blood Blow, Velocity and Heart Function on Circulating Atrial Natriuretic Peptide and Reninangiotension System

Heart function and plasma atrial natriuretic peptide (ANP).plasma renin actiity(PRA) andangiotensionⅡ(Ang Ⅱ) were examined with echocardiography and radioimmunoassay in patients with dilated cardiomyopathy(DCM),