Method for observing Beauveria bassiana colonization in plants.

Microbes interact in a multitude of ways with host plants, can dwell as endophytes within plants causing no apparent disease, and often provide benefits to their host. Observing microorganism distribution and colonization is a prerequisite for interactive research. To this end, we describe use of fluorescent staining for microorganism labeling and highlight its simplicity, and efficiency. Fluorescein can quickly bind to Beauveria bassiana spores, producing bright green fluorescence that can be observed even inside plant tissues. This method provides an intuitive visual image that can be utilised for subsequent data acquisition and statistical analysis.•Our protocol depends on binding of fluorescein diacetate (FDA) specifically to microorganisms. The fungus hydrolyses and metabolises FDA in cells to produce bright green fluorescent products. This fluorescent signal can easily penetrate plant epidermis and be detected by fluorescence microscopy.•FDA, which itself does not emit light, will emit a fluorescent signal only when combined with B. bassiana. Concomitant genetic testing of the fungal ITS confirmed the high level of the fluorescent staining method for detection of B. bassiana.•Compared with the previous green fluorescent protein (GFP) labeling methods, this protocol improved the labeling efficiency of microorganisms and simplifies the process.

A method for evaluating the toxicity interaction of binary mixtures.

Improving the formulation of biological insecticides for greater efficiency and competitiveness is of particular importance with respect to the successful application of these agents in the field. In this regard, mixing different agents is known to be an effective strategy for enhancing practical pest control. However, traditional chemical-based control strategies have significant limitations that compromise control effectiveness, as effects tend to become increasingly dose-dependent over time. To overcome such limitations, and thereby ensure the continuous effective control of insect pests, we herein assessed the efficacy of binary mixtures of biological agents, with the aim of establishing an optimal ratio. The optimized mixture showed a significantly higher insecticidal effect, whereby biological pest control was considerably enhanced.•We combined Beauveria bassiana spores with azadirachtin, a chemical pesticide of botanical origin, and by assessing the efficacy of these two elements combined in different ratios, obtained an optimal formula. Additionally, we evaluated the compatibility between the two elements, and then assessed potential synergistic effects, as determined by the co-toxicity coefficient.•This protocol is dependent on the results of bioassays. It is not only suitable for combining fungal and chemical agents for the purposes of biocontrol but is also applicable to a variety of biological pesticides.•The evaluated binary mixture showed enhanced effective insect control. This approach will contribute to reducing applied dosages, thereby promoting efficient pest control and reducing application costs.

Safety and effectiveness of a Tai Chi-based cardiac rehabilitation programme for chronic coronary syndrom patients: study protocol for a randomised controlled trial.

INTRODUCTION: Preliminary evidence from clinical observations suggests that Tai Chi exercise may offer potential benefits for patients with chronic coronary syndrom (CCS). However, the advantages for CCS patients to practice Tai Chi exercise as rehabilitation have not been rigorously tested and there is a lack of consensus on its benefits. This study aims to develop an innovative Tai Chi Cardiac Rehabilitation Program (TCCRP) for CCS patients and to assess the efficacy, safety and acceptability of the programme. METHODS AND ANALYSIS: We propose to conduct a multicentre randomised controlled clinical trial comprising of 150 participants with CCS. The patients will be randomly assigned in a 1:1 ratio into two groups. The intervention group will participate in a supervised TCCRP held three times a week for 3 months. The control group will receive supervised conventional exercise rehabilitation held three times a week for 3 months. The primary and secondary outcomes will be assessed at baseline, 1 month, 3 months after intervention and after an additional 3-month follow-up period. Primary outcome measures will include a score of 36-Item Short Form Survey and Chinese Perceived Stress Scale. The secondary outcome measures will include body composition, cardiopulmonary exercise test, respiratory muscle function, locomotor skills, echocardiogram, New York Heart Association classification, heart rate recovery time and laboratory examination. Other measures also include Seattle Angina Scale, Pittsburgh Sleep Quality Index, Patient Health Questionnaire-9, Generalized Anxiety Disorder-7 and Berg Balance Scale. All adverse events will be recorded and analysed. ETHICS AND DISSEMINATION: This study conforms to the principles of the Declaration of Helsinki and relevant ethical guidelines. Ethical approval has been obtained from the Ethics Committee of Chinese People's Libration Army General Hospital (approval number: S2019-060-02). Findings from this study will be published and presented at conferences for widespread dissemination of the results. TRIAL REGISTRATION NUMBER: NCT03936504.