Molecular Mechanism of Resistance to Alternaria alternata Apple Pathotype in Apple by Alternative Splicing of Transcription Factor MdMYB6-like.

As a fruit tree with great economic value, apple is widely cultivated in China. However, apple leaf spot disease causes significant damage to apple quality and economic value. In our study, we found that MdMYB6-like is a transcription factor without auto-activation activity and with three alternative spliced variants. Among them, MdMYB6-like-ß responded positively to the pathogen infection. Overexpression of MdMYB6-like-ß increased the lignin content of leaves and improved the pathogenic resistance of apple flesh callus. In addition, all three alternative spliced variants of MdMYB6-like could bind to the promoter of MdBGLU H. Therefore, we believe that MdMYB6-like plays an important role in the infection process of the pathogen and lays a solid foundation for breeding disease-resistant cultivars of apple in the future.

Screening visual environment impact factors and the restorative effect of four visual environment components in large-space alternative care facilities.

Alternative care facilities (ACFs) based on large-space public buildings were widely used early at the start of the coronavirus disease 2019 (COVID-19) pandemic. However, studies have shown that the indoor spatial environment of ACFs can significantly induce mental health problems among users. Thus, this study hypothesizes that improving the visual environment in the interiors of large-space ACFs may reduce mental health problems among users. To verify this hypothesis, this study used critical analysis to screen the influencing factors and used analytic hierarchy process analysis to determine the weights. Particularly, the analyses were based on ACF research in Wuhan and questionnaire surveys of patients with experience using ACFs. Subsequently, virtual reality experiments were conducted to measure physiological indicators and subjective questionnaire collection based on the orthogonal experimental design of the four screened visual environment components. The results revealed the following related to large-space ACFs: 1) Lifestyle support was the most dominant patient requirement and preference for the visual environment. 2) The visual environment can influence the participants' efficiency of psychological stress relief, emotional regulation, and subjective perception. 3) Different design characteristics of the four visual environment components were causally related to restorative effects. To the best of our knowledge, this is the first study analyzing patients' preferences and psychological needs for the visual environment of large-space ACFs and combining subjective and objective measures to investigate the restorative effects of the visual environment. Improving the quality of the visual environment in large-space ACFs presents an effective intervention for alleviating the psychological problems of admitted patients.

Comparative Analysis of Alternative Splicing in Two Contrasting Apple Cultivars Defense against Alternaria alternata Apple Pathotype Infection.

Alternaria blotch disease, caused by the Alternaria alternata apple pathotype (A. alternata AP), is one of the most serious fungal diseases in apples. Alternative splicing (AS), one of the pivotal post-transcriptional regulatory mechanisms, plays essential roles in various disease resistance responses. Here, we performed RNA-Seq for two apple cultivars (resistant cultivar 'Jonathan' (J) and susceptible cultivar 'Starking Delicious' (SD)) infected by A. alternata AP to further investigate their AS divergence. In total, 1454, 1780, 1367 and 1698 specifically regulated differential alternative splicing (DAS) events were detected in J36, J72, SD36 and SD72 groups, respectively. Retained intron (RI) was the dominant AS pattern. Conformably, 642, 764, 585 and 742 uniquely regulated differentially spliced genes (DSGs) were found during A. alternata AP infection. Comparative analysis of AS genes in differential splicing and expression levels suggested that only a small proportion of DSGs overlapped with differentially expressed genes (DEGs). Gene ontology (GO) enrichment analysis demonstrated that the DSGs were significantly enriched at multiple levels of gene expression regulation. Briefly, the specific AS was triggered in apple defense against A. alternata AP. Therefore, this study facilitates our understanding on the roles of AS regulation in response to A. alternata AP infection in apples.

Quantification of human embryonic ζ-globin chains in Southeast Asian deletion (--SEA) carriers.

AIMS: Reactivation of embryonic ζ-globin is a promising strategy for genetic treatment of α-thalassaemia. However, quantification of ζ-globin as a quantitative trait in α-thalassaemia carriers and patients remains incompletely understood. In this study, we aimed to set up a reliable approach for the quantification of ζ-globin in α-thalassaemia carriers, followed by a population study to investigate its expression patterns. METHODS: ζ-globin was purified as monomers from cord blood haemolysate of a Hb Bart's fetus, followed by absolute protein quantification, which was then tested by in-house ELISA system and introduced as protein standard. It was then used for large-scale quantification in peripheral blood samples from 6179 individuals. Finally, liquid chromatography-tandem mass spectrometry (LC-MS/MS) introduced as an independent validating approach by measuring ζ-globin expression in a second cohort of 141-SEA/αα carriers. RESULTS: The ELISA system was proved sensitive in distinguishing individuals with varied extent of ζ-globin. Large scale quantitative study of this --SEA/αα carrier cohort indicated the high diversity of ζ-globin expression ranging from 0.00155 g/L to 1.48778 g/L. Significant positive correlation between ELISA and LC-MS/MS (R=0.400, p<0.001) was observed and it is more sensitive in distinguishing the samples with extreme expression of ζ-globin (R=0.650, p<0.001). CONCLUSION: Our study has reported reliable approaches for the quantification of ζ-globin and presented the expression patterns of ζ-globin among the --SEA/αα carrier population, which might lay a foundation on subsequent genotype-phenotype studies on mechanisms of delayed haemoglobin switch in α-thalassaemia.

Factors and optimizations of healthcare workers' perception in alternative care facilities.

Background: Diverse measures have been carried out worldwide to establish Alternative Care Facilities (ACFs) for different ends, such as receiving, curing or isolating patients, aiming to cope with tremendous shock in the urban medical system during the early passage of the COVID-19 epidemic. Healthcare workers always felt anxious and stressed during multiple major public health emergencies in medical facilities. Some active measures to improve healthcare workers' perceptions, such as temporary training, workflow improvement, and supplementary facilities, were proved insufficient in several past public health emergencies. Therefore, this study aims to analyze the contributing factors of the healthcare workers' perceptions of the ACFs in this pandemic, which can help find an innovative path to ensure their health, well-being and work efficiency. Method: This paper conducted semi-structured in-depth interviews with the world's first batch of healthcare workers who have worked in ACFs through a qualitative study based on Grounded Theory. The healthcare workers interviewed from Heilongjiang, Shandong, Fujian, and Hubei provinces, have worked in one of the four different ACFs built in Wuhan. The results are obtained through the three-level codes and analyses of the interview recordings. Results: The factors affecting the perception of healthcare workers in ACFs during the epidemic situation can be summarized into five major categories: individual characteristics, organization management, facilities and equipment, space design, and internal environment. The five major categories affecting the composition of perception can be further divided into endogenous and exogenous factors, which jointly affect the perception of healthcare workers in ACFs. Among them, individual characteristics belong to endogenous factors, which are the primary conditions, while other categories belong to exogenous factors, which are the decisive conditions. Conclusion: This paper clarifies factors affecting the perception of healthcare workers in ACFs and analyzes the mechanism of each factor. It is posited that the passive strategies are a promising solution to protect healthcare workers' health, improve their work efficiency, and help reduce the operation stress of ACFs. We should train multidisciplinary professionals for future healthcare and enhance collaborations between healthcare workers and engineers. To sum up, this paper broadens new horizons for future research on the optimization of ACFs and finds new paths for alleviating healthcare workers' adverse perceptions of ACFs.

Long non­coding RNA LINC00312 regulates breast cancer progression through the miR­9/CDH1 axis.

Long non­coding RNAs (lncRNAs) are important mediators of the initiation and progression of tumors, including breast cancer (BC). The exact role of long intergenic non­coding RNA 00312 (LINC00312) in BC and its mechanism of action have not been reported to date. In the present study, LINC00312 was found to be downregulated in human BC tissues and cell lines by RT­qPCR. The findings of a functional study indicated that overexpression of LINC00312 suppressed the proliferation, colony forming ability, migration and invasiveness of BC cell lines. Mechanistically, LINC00312 was found to induce suppression of cell migration and invasion by directly binding to miR­9. Overexpression of LINC00312 increased the expression of cadherin 1 (CDH1), a direct target of miR­9, and decreased the expression of vimentin (VIM), a major cytoskeletal component of mesenchymal cells as determined by western blot analysis. miR­9 partly abrogated the upregulation of CDH1 and downregulation of VIM induced by LINC00312. Taken together, the results of the present study indicate a role for the LINC00312/miR­9/CDH1 axis in the progression of BC, and suggest a novel lncRNA­based diagnostic biomarker or therapeutic target for BC.

[The application of multiplex ligation-dependent probe amplification technology in diagnosis and prenatal diagnosis of α-thalassemia].

OBJECTIVE: To investigate the multiplex ligation-dependent probe amplification (MLPA) technology in the detection of gene deletion and prenatal diagnosis of α-thalassaemia. METHODS: Phenotypes were analyzed by whole blood cell counting and hemoglobin component detection of peripheral blood samples from the subjects. The gene deletions and point mutations of α- thalassaemia were detected with regular gap-PCR and reverse dot blot (RDB) method. At last, the MLPA method was applied for detection of α-globin gene deletion. All the prenatal diagnosis samples were detected with both gap-PCR and MLPA method. RESULTS: α-thalassaemia phenotype was found in 75 samples from 1256 (628 couples) peripheral blood samples for pre-pregnancy or prenatal thalassemia gene screening. Among them, 71 samples carrying α-gene mutations and consistent with phenotypes were detected by routine methods. In the other 3 samples with no α-gene mutations detected and 1 sample with HbH phenotype but genotype of ï¹£α(4.2)/αα were analyzed by MLPA and found each one samples of whole α-globin gene cluster deletion, respectively. Seventeen high risk couples were screened. Among the 17 prenatal diagnosis samples, 2 villus samples contaminated by exogenous DNA were confirmed by MLPA method. CONCLUSION: MLPA is an effective complement for α-thalassaemia gene deletion detection. The molecular diagnosis strategy and process of gap-PCR combined with MLPA for α- thalassaemia gene deletion detection can prevent the missing of gene deletion, and false-positive or false-negative misdiagnosis of α-thalassaemia in prenatal diagnosis.