Sets of serum exosomal microRNAs as candidate diagnostic biomarkers for Kawasaki disease.

Although Kawasaki disease is the main cause of acquired heart disease in children, no diagnostic biomarkers are available. We aimed to identify candidate biomarkers for diagnosing Kawasaki disease using serum exosomal microRNAs (miRNAs). Using frozen serum samples from a biobank, high-throughput microarray technologies, two-stage real-time quantitative PCR, and a self-referencing strategy for data normalization, we narrowed down the list of biomarker candidates to a set of 4 miRNAs. We further validated the diagnostic capabilities of the identified miRNAs (namely, CT(miR-1246)-CT(miR-4436b-5p) and CT(miR-197-3p)-CT(miR-671-5p)) in 79 samples from two hospitals. We found that this 4-miRNA set could distinguish KD patients from other febrile patients as well as from healthy individuals in a single pass, with a minimal rate of false positives and negatives. We thus propose, for the first time, that serum exosomal miRNAs represent candidate diagnostic biomarkers for Kawasaki disease. Additionally, we describe an effective strategy of screening for biomarkers of complex diseases even when little mechanistic knowledge is available.

A novel NKX2.6 mutation associated with congenital ventricular septal defect.

Congenital heart disease (CHD) is the most common birth defect and is the most prevalent non-infectious cause of infant death. Aggregating evidence demonstrates that genetic defects are involved in the pathogenesis of CHD. However, CHD is genetically heterogeneous and the genetic determinants for CHD in an overwhelming majority of patients remain unknown. In this study, the coding regions and splice junctions of the NKX2.6 gene, which encodes a homeodomain transcription factor crucial for cardiovascular development, were sequenced in 210 unrelated CHD patients. As a result, a novel heterozygous NKX2.6 mutation, p.K152Q, was identified in an index patient with ventricular septal defect (VSD). Genetic analysis of the proband's available family members showed that the mutation cosegregated with VSD transmitted as an autosomal dominant trait with complete penetrance. The missense mutation was absent in 400 control chromosomes and the altered amino acid was completely conserved evolutionarily across species. Due to unknown transcriptional targets of NKX2.6, the functional characteristics of the identified mutation at transcriptional activity were analyzed by using NKX2.5 as a surrogate. Alignment between human NKX2.6 and NKX2.5 proteins displayed that K152Q-mutant NKX2.6 was equivalent to K158Q-mutant NKX2.5, and introduction of K158Q into NKX2.5 significantly reduced its transcriptional activating function when compared with its wild-type counterpart. This study firstly links NKX2.6 loss-of-function mutation with increased susceptibility to isolated VSD, providing novel insight into the molecular mechanism underpinning VSD and contributing to the development of new preventive and therapeutic strategies for this common form of CHD.

Potent cationic inhibitors of West Nile virus NS2B/NS3 protease with serum stability, cell permeability and antiviral activity.

West Nile virus (WNV) has spread rapidly around the globe, efficiently crossing species from migrating birds into humans and other mammals. The viral protease NS2B-NS3 is important for WNV replication and recognizes dibasic substrate sequences common to other flaviviral proteases but different from most mammalian proteases. Potent inhibitors of WNV protease with antiviral activity have been elusive to date. We report the smallest and most potent inhibitors known for this enzyme, cationic tripeptides with nonpeptidic caps at the N-terminus and aldehyde at the C-terminus. One of these, compound 3 ( Ki = 9 nM) is stable in serum (>90% intact after 3 h, 37 degrees C), cell permeable, and shows antiviral activity (IC 50 1.6 microM) without cytotoxicity (IC 50 >400 microM), thereby validating the approach of inhibiting WNV protease to suppress WNV replication.

[Isolation and identification of imazethapyr degradable bacteria and its degradation characteristics].

One strain of imazethapyr degradable bacteria was isolated from the mixture of imazethapyr production factory's sludge and the soil that was contaminated with imazethapyr for a long time. The strain could degrade imazethapyr more than 90% within 72 h when it grew in 500 mg/L imazethapyr condition. When pH was 5, the bacteria could degrade all the imazethapyr of 500 mg/L in 72 h, but when pH was 8 and 9, only 50% imazethapyr was degraded which showed acid condition was more compatible than that of alkalescency for this bacteria growing. The higher degradation ratio at 25 degrees C and 30 degrees C was observed and the optimum condition for the bacteria to degrade imazethapyr was pH = 5 and 25 degrees C. Characterizing by physiological and biochemical properties as well as 16S rRNA sequence analysis, the strain is related and shared characteristics of the genus Alcaligenes sp.

[Non-dilated coronary artery lesion and myocardial perfusion in children with Kawasaki disease: analysis of 43 cases].

OBJECTIVE: The study was designed to investigate the impact of non-dilated coronary artery wall lesion on myocardial perfusion. METHODS: Doppler tissue image (DTI) was used to measure regional ventricular wall motion in 43 Kawasaki children with non-dilated coronary arterial wall echocardiographic abnormalities (rough intima and arterial wall thickening) detected by two-dimensional echocardiography (2DE) at acute phase. A total of 31 cases who had both non-dilated coronary lesion and lowered ventricular wall motion velocity at subacute and convalescence phase underwent submaximal exercise single photon emitting computerized tomography (SPECT) for the evaluation of myocardial perfusion. RESULTS: In 43 cases of Kawasaki disease with non-dilated coronary arterial wall abnormalities, 36 cases (83.7%) still had such lesions at subacute phase and 32 (74.4%)at convalescence. At the same time, lowered regional ventricular wall motion (RVWM) was found in 34 cases at subacute phase and in 31 cases at convalescence. DTI and 2DE had a very good correlation in the detection of such abnormalities (chi(c)2 = 9.64, P < 0.01 in subacute period, and chi(c)2 = 7.14, P < 0.01 in convalescence). In 31 cases accepting SPECT, 17 were positive. A total of 22 ischemic regions were detected. Eighteen out of 22 cases having ischemic regions had abnormal RVWM on DTI. SPECT ischemic regions were significantly in accordance with lowered RVWM in ventricular septum and anterior wall (chi(c)2=5.07 and 7.48, P < 0.05 and P < 0.01, respectively) noted in DTI. CONCLUSION: Non-dilated coronary arterial wall abnormality is one of the forms of coronary artery wall lesions which could reduce myocardial flow perfusion. Its clinical significance is worthy of attention.