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A nanopore counter for highly sensitive DNA methylation evaluation was developed by relating the methylation level to the concentration of the generated fixed-length DNA duplex amplicons, which achieved a limit of detection of 0.61 aM.

Objectives To investigated the clinical characteristics and prognosis of hypothyroid myopathy Methods The clinical characteristics were analyzed in 10 patients diagnosed with polymyositis-like hypothyroid myopathy. Symptoms and indicators at baseline and after levothyroxine replacement therapy were compared. The correlations of muscle enzyme levels with thyroid function were analyzed. Results A total of 10 patients, including 3 males and 7 females, were enrolled. The average onset age was 46.1 ± 10.3 years and duration of disease was 2.1 ± 0.9 months. The first manifestation comprised proximal muscle weakness and myalgia or arthralgia. Liver dysfunction, hyperlipidemia, muscle enzyme increased, pericardial effusion, pleural effusion, and fatty liver was common. All patients were Hashimoto's thyroiditis and negative for myositis antibodies. Pathological muscle biopsies revealed myxoid degeneration and muscle atrophy. The proportion of patients who experienced proximal muscle weakness, myalgia, and/or arthralgia, ALT increased, AST increased, CHOL increased, TG increased, CK increased , CK-MB increased, LDH increased, fT3 decreased, and fT4 decreased were significantly lower after treatment(all PPPConclusion Hypothyroid myopathy was more common in female. Most patients had liver dysfunction, hyperlipidemia and abnormal muscle enzyme levels, serous cavity effusion and fatty liver. Electromyography and muscle pathology had no specific alterations. The prognosis was well after levothyroxine replacement therapy.

Solid-state nanopores have been proven as a powerful platform for label-free single-molecule analysis. However, due to its relatively low resolution and selectivity, developing biosensors with good translocation signals faces two significant challenges: (1) small-sized chemical or biological targets show difficulty in producing recognizable translocation signals because of their weak interaction with the nanopore and (2) protein interferents that widely exist in biological samples or buffers would considerably deteriorate the noise level of the nanopore, submerging the translocation signal. Herein, we demonstrate an effective way to overcome both the challenges. DNA cubes were used as signal transducers that can achieve an ultra-high (50 : 1) signal-to-noise ratio (SNR) translocation signal, which is maintained even in protein interferent-rich buffers. A sensing strategy was constructed

Lysine lipoylation, a highly conserved lysine post-translational modification, plays a critical role in regulating cell metabolism. The catalytic activity of a number of vital metabolic proteins, such as pyruvate dehydrogenase (PDH), depends on lysine lipoylation. Despite its important roles, the detailed biological regulatory mechanism of lysine lipoylation remains largely unexplored. Herein we designed a powerful affinity-based probe, KPlip, to interrogate the interactions of lipoylated peptide/proteins under native cellular environment. Large-scale chemical proteomics analysis revealed a number of binding proteins of KPlip, including sirtuin 2 (Sirt2), an NAD+-dependent protein deacylase. To explore the potential activity of Sirt2 toward lysine lipoylation, we designed a single-step fluorogenic probe, KTlip, which reports delipoylation activity in a continuous manner. The results showed that Sirt2 led to significant delipoylation of KTlip, displaying up to a 60-fold fluorescence increase in the assay. Further kinetic experiments with different peptide substrates revealed that Sirt2 can catalyze the delipoylation of peptide (DLAT-PDH, K259) with a remarkable catalytic efficiency (kcat/Km) of 3.26 × 103 s-1 M-1. The activity is about 400-fold higher than that of Sirt4, the only mammalian enzyme with known delipoylation activity. Furthermore, overexpression and silencing experiments demonstrated that Sirt2 regulates the lipoylation level and the activity of endogenous PDH, thus unequivocally confirming that PDH is a genuine physiological substrate of Sirt2. Using our chemical probes, we have successfully established the relationship between Sirt2 and lysine lipoylation in living cells for the first time. We envision that such chemical probes will serve as useful tools for delineating the roles of lysine lipoylation in biology and diseases.

Histone deacetylases (HDACs) play important roles in regulating various physiological and pathological processes. Developing fluorescent probes capable of detecting HDAC activity can help further elucidate the roles of HDACs in biology. In this study, we first developed a set of activity-based fluorescent probes by incorporating the Kac residue and the O-NBD group. Upon enzymatic removal of the acetyl group in the Kac residue, the released free amine reacted intramolecularly with the O-NBD moiety, resulting in turn-on fluorescence. These designed probes are capable of detecting HDAC activity in a continuous fashion, thereby eliminating the extra step of fluorescence development. Remarkably, the amount of turn-on fluorescence can be as high as 50-fold, which is superior to the existing one-step HDAC fluorescent probes. Inhibition experiments further proved that the probes can serve as useful tools for screening HDAC inhibitors. Building on these results, we moved on and designed a dual-purpose fluorescent probe by introducing a diazirine photo-cross-linker into the probe. The resulting probe was not only capable of reporting enzymatic activity but also able to directly identify and capture the protein targets from the complex cellular environment. By combining a fluorometric method and in-gel fluorescence scanning technique, we found that epigenetic readers and erasers can be readily identified and differentiated using a single probe. This is not achievable with traditional photoaffinity probes. In light of the prominent properties and the diverse functions of this newly developed probe, we envision that it can provide a robust tool for functional analysis of HDACs and facilitate future drug discovery in epigenetics.

Currently, villages “besieged with garbage” have become a serious problem in rural areas of China. Separation of rural residential solid waste (RRSW) is one of the main strategies for waste reduction. Although previous studies have analyzed the social and psychological motivations of residents’ separation intention for municipal solid waste (MSW), little attention has been paid to the situation in rural areas. This paper investigates key factors influencing rural residents’ separation intention, as well as analyzing the moderating effects of perceived policy effectiveness on the relationship between the determinants and the intention, using survey data of 538 rural residents in the province of Sichuan in China. The results show that all the proposed key factors influence the separation intention significantly. Furthermore, the policies were divided into two types and the moderating effects were tested for each type. The results show that the perceived effectiveness of both the inducement policy and the capacity building policy moderated the relationship between attitude and separation intention positively, while the perceived effectiveness of the inducement policy moderated the relationship between subjective norms and intention negatively. The findings provide insightful information for policymakers to design effective RRSW separation policies.

To compare the difference of clinical efficacy on sleeping disorder in the children with encephalopathy between the combined therapy of acupuncture at head points and seed-pressure at ear points and the simple acupuncture at head points.Thirty cases of sleeping disorder induced by encephalopathy werei randomized into an observation group and a control group, 15 cases in each one. In the observation group, the combined therapy of acupuncture at head points and seed-pressure at ear points was adopted. The head points in cluded Sishencong (EX-HN 1), Shenting (GV 24) and Benshen (GB 13). The ear points were the positive reactive sites in the cymba and cavum conchae. In the control group, acupuncture was applied simply to the acupoints on the head. The treatment was given once on every Tuesday and Friday a week separately, 30 min each time. Totally, 16 treatments were required. Children's sleeping habit questionnaire (CSHQ) was used to observe the sleep improvements and the efficacy in the patients of the two groups.In the observation group, the results of sleep resistance, sleep anxiety, night sleep wake, parasomnias, sleep dyspnea, daytime somnolence and the total score after treatment were all improved apparently as compared with those before treatment (all P0. 05). In the control group, the results of night sleep wake, parasomnias, daytime somnolence and the total score after treatment were improved apparently than those before treatment (all P0. 05). In the observation group, the results of sleep resistance, sleep dyspnea and the total score after treatment were better than those in the control group (all P0. 05) and the scores of sleep anxiety and daytime somnolence in the control group were better than those in the observation group after treatment (both P0. 05).The combined therapy of acupuncture at head points and seed-pressure at the positive reactive sites in the cymba and cavum conchae achieves the superior efficacy on sleep resistance and sleep dyspnea as compared with the simple acupuncture. The efficacy of simple acupuncture is more satisfactory on sleep anxiety and daytime somnolence.

A real‐time environment information acquisition system is essential if models and vegetable‐crop information are to be integrated with a greenhouse management expert system for good decision making. Our greenhouse management expert system has four functional modules: (1) cultivation techniques, (2) pest and disease diagnosis and prevention, (3) nutrient deficiency diagnosis and fertilisation, and (4) environment control. The hardware and software of the environment information acquisition system are incorporated into the expert system, which also offers a multi‐interface for sensors and is easily extended and maintained. Implementation was accomplished with the whole system to ensure its reliability and applicability for expert system, on‐line decision making. The results show that a dynamic integration of a greenhouse management system and an environment information acquisition system can supply sufficient information for good control strategies and for decision‐support.

Objective: To study the nature of preoperative nursing care of a Laryngoscope facilitated vocal polyp excision procedure using general anesthesia. Methods: Clinical data from 238 cases of the aforementioned laryngoscope procedure from May 2012 to May 2014 in our hospital’s Otolaryngology department was collected. Psychological nursing was administered including preoperative preparation, postoperative monitoring, close observation of vital signs, post-operative nursing, diet, care, and also enhanced nursing skills such as discharge guidance. Results: 238 cases of vocal cord polyp patients were all completely cured and discharged, there was no postoperative infection and other complications. After reviewing the condition of the vocal cords, primarily if the sutures at the incision are fine, there is apparent improvement. Conclusion: The laryngoscope facilitated vocal polyp excision under general anesthesia procedure can deliver satisfying results when coupled with close patient observation and improved understanding of diseases by the preoperative nurses.