Effects of Temperature, Moisture, and Ultraviolet Light on Germination, Infection, and Survival of Gymnosporangium yamadae Teliospores and Basidiospores.

Apple rust caused by Gymnosporangium yamadae is a significant disease in China's main apple production areas. We evaluated the effects of temperature, moisture, and ultraviolet (UV) light on the germination, infection, and survival of teliospore horns and basidiospores under artificially controlled environmental conditions. The temperature required for the germination and infection of teliospores and basidiospores of G. yamadae ranged from 5 to 25°C, with an optimum temperature of approximately 17°C. The teliospore horns germinated after soaking in distilled water for 5 min and required at least 2.3 h of development to produce basidiospores under the most favorable conditions. The basidiospores germinated only in free water and produced germ tubes 0.8 h after being placed in the water. The half-life of the basidiospore was 72.5 h in the dark and only 9.5 h when exposed to intense UV light. The basidiospores inoculated on the host leaves required at least 2.3 h of water exposure to cause rust lesions. A revised Weibull model could describe the relationships between the germination and infection of teliospore horns and basidiospores with temperature and wetness duration. Collectively, these results can serve as a valuable guide for developing a model to predict future apple rust epidemics and establish a method for effective control strategies.

Detection of Cytb Point Mutation (G143A) that Confers High-Level Resistance to Pyraclostrobin in Glomerella cingulata Using LAMP Method.

Glomerella leaf spot (GLS) caused by Glomerella cingulata is a newly emerging disease that results in severe defoliation and fruit spots in apples. In China, the compound of pyraclostrobin and tebuconazole was registered to control GLS in 2018 and has achieved excellent control efficiency. In this study, we showed that the high-level resistant isolates of G. cingulata to pyraclostrobin, caused by the point mutation at codon 143 (GGT→GCT, G143A) in the cytochrome b gene, has appeared in apple orchards in Shandong Province in 2020, and the resistance frequency was 4.8%. Based on the genotype of the resistant isolates, we developed a loop-mediated isothermal amplification (LAMP) assay for detection of the pyraclostrobin resistance. The LAMP assay was demonstrated to have good specificity, sensitivity, and repeatability, and it exhibited high accuracy in detecting pyraclostrobin resistance in the field. This study reported the resistance status of GLS to pyraclostrobin in Shandong Province and developed a molecular tool for the detection of pyraclostrobin resistance, which is of practical significance for the scientific control of GLS.

The Evaluation Indexes Suitable for Nonhuman Primates can be Extracted from Clinical Consciousness Disorder Assessment Scales: A Hypothesis.

BACKGROUND: Currently, case studies or clinical trials in different patient populations remain the main resource underlying the understanding of disorder of consciousness (DoC). This provides a low efficacy for the derivation of data and the implementation of associated controlled experimental designs. Preclinical models provide precise controls, reduced variability, rich data output and limited ethical complexity. Nonhuman primates are suitable model animals for disorders of consciousness due to their brain structure being very similar to that of humans. Behavioral tests remain the primary standard for assessing the consciousness status of humans. However, there is currently no behavioral assessment scale available for evaluation of the state of consciousness disorder in nonhuman primates. This presents a significant challenge for the establishment of different models of consciousness disorder. Therefore, there is considerable motivation to focus on the development of a proper tool for assessment of the state of consciousness associated with nonhuman primate models that are based on clinically common consciousness assessment scales. METHODS: It is assumed that the Delphi and level analysis methods based on clinical consciousness disorder assessment scales may provide an effective way to select and include assessment indexes for levels of consciousness in nonhuman primates. RESULTS: 8 first-level indicators with 41 second-level indexes were selected preliminary as a pool of evaluation entries of state of consciousness of nonhuman primates. CONCLUSIONS: It may be practicable to extract appropriate indicators for non-human primates from the clinical consciousness disorder assessment scales. Besides, a combination of Delphi method, behavioral analysis, electroencephalography, neuroimaging (such as positron emission tomography-computed tomography) and functional magnetic resonance imaging is necessary to test the reliability and validity of the novel scale reported here.

A randomized controlled trial to explore the efficacy and safety of transcranial direct current stimulation on patients with post-stroke fatigue.

BACKGROUND: Post-stroke fatigue seriously affects the quality of life for stroke patients. There is no effective treatment at present. transcranial direct current stimulation (tDCS) is a non-invasive brain stimulation which may have therapeutic effect on post-stroke fatigue. This study will explore about this. METHOD: A total of 60 patients with post-stroke fatigue were randomly divided into the control group and the treatment group with 30 patients each by minimization randomization. Both groups received basic treatment and conventional rehabilitation. In the treatment group, patients were treated with active tDCS, while in the control group, sham tDCS. Both active and sham tDCS were administered 6 times a week for 4 weeks. Before and after the trial, the Fatigue Severity Scale (FSS), Fugl-Meyer Assessment (FMA) and Modified Barthel Index (MBI) were evaluated and analyzed. And comparisons were made among groups. And there were an 8-week follow-up after the intervention. RESULT: Before the intervention, there were no significant differences in baseline data and assessment scores between the groups (P > 0.05). After 4 weeks of intervention, FSS scores in the treatment group were significantly lower than those in the control group (P = 0.012), and FMA and BMI scores were significantly higher than those in the control group (P < 0.05). There was no significant change in FSS scores after 8 months of follow-up (P > 0.05). DISCUSSION: TDCS is a safe treatment that can effectively reduce the degree of fatigue after stroke, improve the motor function and daily activity ability of patients after stroke, and the efficacy is better than only routine rehabilitation training. TRIAL REGISTRATION NUMBER: Chinese Clinical Trial Registry, ChiCTR2000031120. Registered on March 22, 2020.

Progression of Symptoms Caused by Botryosphaeria dothidea on Apple Branches.

Until recently, the causal agent of Botryosphaeria canker was assumed to differ from that causing ring rot on fruit and warts on branches on apple trees in China and East Asia. However, recent research documented that Botryosphaeria dothidea caused both disease symptoms on apple. Inoculations with strains isolated from cankers and warts on branches were conducted to investigate symptom progression caused by B. dothidea and conditions inducing the two symptom types. The results confirmed that both cankers and warts are caused by B. dothidea. Warts are the results of hyperplasia and suberization of bark tissues induced by fungal infection, whereas cankers result from the rapid growth of hyphae from inside warts, lenticels, or wounds. Resistance to B. dothidea exists in living apple branches. When a living branch is infected via lenticels, the pathogen induces proliferation and suberization of cortical cells that restricts the growth and expansion of the hyphae, leading to warts. However, under certain stress conditions such as drought, the hyphae inside host tissues expand rapidly and kill cortical cells, leading to canker development. Host resistance may recover during active growth periods, which suppresses or even stops rapid expansion of the hyphae, leading to the intermediate symptom of canker warts. Abiotic factors, such as drought or high temperature in early spring, can result in rapid extension of colonized hyphae in branches and conversion of warts to cankers. Preventing this transition can be an important measure in managing Botryosphaeria canker on apple.

Effects of Temperature and Moisture on Conidia Germination, Infection, and Acervulus Formation of the Apple Marssonina Leaf Blotch Pathogen (Diplocarpon mali) in China.

Apple Marssonina leaf blotch (AMLB; Diplocarpon mali) is a severe disease of apple that mainly causes premature leaf defoliation in many apple growing areas worldwide. AMLB epidemic development is closely related to temperature and rainfall. In this study, the effects of temperature and moisture on conidium germination, infection on leaves, and acervulus production were investigated under controlled environments. The temperature required for conidium germination and infection ranged from 5 to 30°C, with the optimum at approximately 23°C. The temperature required for acervulus formation was slightly higher, with the optimum at 24.6°C. Wetness was needed in order for conidia to germinate and infect; only a few conidia germinated at 100% RH. However, lesions can produce acervuli in dry conditions. The minimum duration of leaf wetness required for conidia to complete the entire infection process was 14, 8, 4, and 6 h at 10, 15, 20, and 25°C, respectively. A model describing the effect of temperature and leaf wetness duration was built. The model estimated that the optimum temperature for conidial infection was 22.6°C and the minimum wetness duration required was 4.8 h. This model can be used to forecast D. mali conidial infection to assist in disease management in commercial apple production.

Effect of Rainfall and Temperature on Perithecium Production of Botryosphaeria dothidea on Cankered Apple Branches.

Botryosphaeria dothidea is a fungal pathogen causing canker, dieback, and fruit rot of apple trees worldwide. Ascospores are an important source of inoculum of Botryosphaeria canker in China. Experiments were conducted under both controlled and natural conditions to study perithecium formation in relation to environmental conditions. Perithecia of B. dothidea were detected on cankered lesions throughout the apple growing season except in July and in some years including August under natural conditions. On newly formed canker lesions, the first perithecium was detected as early as August, about 1 week after rainfall. Perithecia matured successively, lasting from early August to June of the next year, with a peak in late September or early October. Temperature and rainfall are two key environmental factors affecting perithecium formation. Under controlled conditions, perithecia were produced only on cankered shoots incubated at test temperatures of 20 and 25°C and wetted by >3 days of simulated rainfall per week. The number of perithecia produced on canker lesions increased with the increase in rainfall duration. Perithecia were formed on canker shoots exposed to rainfall only in June, July, and August but not in September. Rainfall of >3 days per week can be used to predict the initial formation of perithecia in the main apple production areas in China to assist disease management.

Effects of Temperature and Moisture on the Infection and Development of Apple Fruit Rot Caused by Phytophthora cactorum.

Phytophthora fruit rot, caused by Phytophthora cactorum, is an important disease of apple in China, often causing more than 50% fruit rot in rainy years. We examined the effects of temperature and moisture on the development of the disease and effects of the variables on zoospore release and germination, infection, and lesion development. In vitro, a temperature range of 5 to 20°C had no significant effects on zoospore release dynamics but did significantly affect the quantities of released zoospores. The largest quantity of zoospores was released at 9.9°C according to a fitted model. Zoosporangia released zoospores within 15 min at the test temperatures (0 to 20°C), which peaked at the fourth hour. Zoospores germinated in vitro, requiring free water, at temperatures from 5 to 35°C. The optimum germination temperature was 25.1°C according to a fitted model. The minimum wetness duration required for zoospores to complete the infection process and induce visible lesions on Fuji fruit was 0.40 h at the optimal temperature of 23.0°C according to the fitted model, whereas observed values were 4.5, 1.5, 0.5, 1.5 and 8.5 h at 10, 15, 20, 25, and 30°C, respectively. The number of zoospore infections on fruit at various temperatures and wetness durations were well fitted by the modified Weibull model; based on the model, the optimal temperature for zoospore infections was 23.0°C. Young apple fruit infected by zoospores developed visible lesions from 10 to 30°C, with a predicted optimum of 23.5°C; no lesions developed at 5 or 35°C. The shortest incubation period of the disease was 4 days. These results can be used to develop disease forecasting models for improved fungicide control.

A evaluation for uncertainty analysis of the verification result of ELISA analyzer(microplate reader) / 中国医学装备

Objective: To analyze various factors that affect the verification result in the verification by using ELISA analyzer and assess the accuracy of verification results of ELISA analyzer. Methods: Through the comparison of verification results and the calculation for the repeatability of measurement standard, the accuracy of standard device was analyzed and verified. Results: Through analyzed the effect of repeatability of operation and standard device, the factors that affected verification results were eliminated. And the accuracy of verification results were guaranteed. Conclusion: In the process of verification for ELISA analyzer, operators should pay attention to the setting of the verification instrument. And the operation regulation can ensure the accuracy and reliability of the measured value of spectrophotometer. Besides, operators also should pay attention to the analysis of verification results and confirm whether the repeatability of the verification results can meet the requirement of practice.

Improvement of medical equipment quality control management module / 医疗卫生装备

Objective To improve medical equipment quality control management module to report medical equipment adverse events,evaluating medical equipment,managing medical metrology certificate and etc.Methods The module with four functions was developed with the original equipment management information system as well as the list structures of eq_measuring and eq_measuring_detailed and Microsoft Visual Studio 2010.Rwesults The system realized the management of adverse events,evaluation,scrapping and certificate,and could be used to master the quality of in-use equipment based on data analysis.Conclusion The system behaves well in stability and usage,and thus is worthy promoting for enhancing medical equipment quality coutrol and management.

Improvement of medical equipment quality control management module / 医疗卫生装备

Objective To improve medical equipment quality control management module to report medical equipment adverse events,evaluating medical equipment,managing medical metrology certificate and etc.Methods The module with four functions was developed with the original equipment management information system as well as the list structures of eq_measuring and eq_measuring_detailed and Microsoft Visual Studio 2010.Rwesults The system realized the management of adverse events,evaluation,scrapping and certificate,and could be used to master the quality of in-use equipment based on data analysis.Conclusion The system behaves well in stability and usage,and thus is worthy promoting for enhancing medical equipment quality coutrol and management.

Effects of Temperature, Humidity, and Wound Age on Valsa mali Infection of Apple Shoot Pruning Wounds.

Valsa canker, caused by Valsa mali, is a destructive disease of apple in China. The pathogen infects apple branches, mainly through pruning wounds, and causes branch and tree death. To determine the conditions required for V. mali infection through pruning wounds and growth within the xylem, pruning wounds on 1- to 4-year-old apple branches were inoculated with conidia in vitro under artificially controlled conditions and in vivo in the orchard. The effects of temperature, wetness duration, and wound age on conidial infection through pruning wounds as well as hyphal growth in the xylem were examined. The results showed that, after invading through pruning wounds, V. mali hyphae grew along xylem vessels, tracheids, and rays, expanding longitudinally and laterally. The hyphae could enter adjacent xylem vessels and tracheids through micropores to form a dense hyphal network. Wetness duration did not exhibit an essential effect on conidial infection from pruning wounds. Conidia spread to pruning wounds with rainwater could infect the xylem without any other extra moisture. Temperature for V. mali conidia infection through pruning wounds and hyphal extension in the xylem ranged from 5 to 35°C, with the optimum at 20°C. Pruning wounds made in late March were susceptible to V. mali infection in March, April, and May; the susceptibility was markedly deceased by June, and the pathogen could barely infect through the pruning wounds in November. The infected pruning wounds began to show symptoms from the spring of the following year. More than half of the observed Valsa canker lesions emerged in the spring of the second year, and new canker twigs were also developed from the inoculations in the spring of the third year. March, April, and May are the critical periods for protecting pruning wounds against infection by V. mali in China, and coating pruning wounds with protective film immediately after pruning is an easy and effective measure to protect the pruning wounds.

Bioactive consistency evaluation of cardiotonic pill using marker genes / 药学学报

The study is aimed to develop a method in evaluation of the bioactive consistency of cardiotonic pill (CP). HepG2 cell line was employed as a biological detector. After treated with CP for 24 h, gene chip and qRT-PCR were used to select mRNAs that can represent the bioactivity of CP. Then similarity between different batches of CP were calculated based on expression levels of marker genes to evaluate the bioactive consistency of CP. Marker genes were selected according to the criteria as follows:① fold change 1.5; ② potential relevance to curative effects; ③ extensive involvement in the cellular functions and clustering analysis categories; ④ dose-dependent effect. A total of 10 genes were selected as bioactive markers of CP. Angular cosine was calculated to evaluate the similarity between two samples. The method was validated using intra-day precision and inter-day precision. Using angular cosine similarity, the intra-day and inter-day precision were 0.4% and 0.6%, respectively. The similarities of 6 batches of CDPs ranged from 0.992 to 0.999, and 1 batch of Compound Danshen Tablet was 0.534. The established method is specific and accurate, and provides comprehensive and objective evaluation of bioactive quality of CDPs. It can also benefit the bioactive consistency evaluation of other compounds in traditional Chinese medicines.

Identification of chemical constituents in Yangxue Qingnao granule by UPLC-Q-TOF/MSE / 药学学报

In order to clarify the chemical constituents in Yangxue Qingnao granule, we established a rapid ultra-performance liquid chromatography/orthogonal acceleration time-of-flight mass spectrometry (UPLCQ- TOF/MSE) method. According to the high resolution MS spectra data, fragmentation ion information and retention time, 142 peaks were identified or tentatively presumed by comparison with reference standards data and literature reports. The herbal sources of these peaks were assigned. The results implied that phenolic acids, alkaloids, anthraquinones, phthalides, monoterpene glycosides were included in the main components of Yangxue Qingnao granule. The method is rapid for systematically elucidation of the constituents of Yangxue Qingnao granule and the results would facilitate the quality control of Yangxue Qingnao granule for safe and efficacious use.

Effects of Temperature, Wetness Duration, and Moisture on the Conidial Germination, Infection, and Disease Incubation Period of Glomerella cingulata.

Glomerella leaf spot (GLS) caused by Glomerella cingulata is a newly emergent disease that results in severe defoliation and fruit spots. Currently, GLS is not effectively controlled in China due to a lack of understanding of its epidemiology. Therefore, the effects of temperature, wetness duration, and moisture on conidial germination, infection, and the disease incubation period of GLS were examined by inoculating cv. Gala apple leaves with a conidial suspension and performing in vitro germination assays. Conidia could germinate and form appressoria at temperatures ranging from 5 to 35°C, with an optimum temperature of 27.6°C. The germination of conidia required free water or a nearly saturated relative humidity, with only a few conidia germinating and forming appressoria when the RH was less than 99%. The conidial germination dynamics at 10, 25, and 30°C were well represented by three logistic models. The infection of cv. Gala apple leaves by conidia occurred at temperatures ranging from 15 to 35°C. The minimum wetness duration required for infection by conidia at different temperatures was described using a polynomial equation, and the lowest minimum wetness duration was 2.76 h, which occurred at 27.6°C according to the polynomial. Successful infection by conidia was represented by the number of lesions per leaf, which increased with extended wetness durations at the conidial infection stage for six tested temperatures, with the exception of 10°C, when the minimum wetness durations were satisfied. The associations of successfully infected conidia with wetness duration at temperatures of 15, 20, 25, and 30°C were described by four logistic models. Conidia infections developed into visible lesions at temperatures ranging from 15 to 30°C, and the shortest incubation period of 2 days was observed at 25°C. These data and models can be used to construct forecasting models and develop effective control systems for Glomerella leaf spot.