Phase separation in DNA double-strand break response.

DNA double-strand break (DSB) is the most dangerous type of DNA damage, which may lead to cell death or oncogenic mutations. Homologous recombination (HR) and nonhomologous end-joining (NHEJ) are two typical DSB repair mechanisms. Recently, many studies have revealed that liquid-liquid phase separation (LLPS) plays a pivotal role in DSB repair and response. Through LLPS, the crucial biomolecules are quickly recruited to damaged sites with a high concentration to ensure DNA repair is conducted quickly and efficiently, which facilitates DSB repair factors activating downstream proteins or transmitting signals. In addition, the dysregulation of the DSB repair factor's phase separation has been reported to promote the development of a variety of diseases. This review not only provides a comprehensive overview of the emerging roles of LLPS in the repair of DSB but also sheds light on the regulatory patterns of phase separation in relation to the DNA damage response (DDR).

Detection of myeloma cell-derived microvesicles: a tool to monitor multiple myeloma load.

The persistence of tumor load in multiple myeloma (MM) lead to relapse in patients achieving complete remission (CR). Appropriate and effective methods of myeloma tumor load monitoring are important for guiding clinical management. This study aimed to clarify the value of microvesicles in monitoring MM tumor load. Microvesicles in bone marrow and peripheral blood were isolated by differential ultracentrifugation and detected by flow cytometry. Western blotting was applied to assess myosin light chain phosphorylation levels. Flow cytometry to detect Ps+CD41a-, Ps+CD41a-CD138+, Ps+CD41a-BCMA+ microvesicles from bone marrow can be used to predict myeloma burden, furthermore, Ps+CD41a- microvesicles may as a potential index to MRD test. Mechanistically, the releasing of microvesicles from MM cell was regulated by Pim-2 Kinase via Phosphorylation of MLC-2 protein.

Qualitative Study on Psychological Experience of Close Contacts with COVID-19 during the Centralized Isolation Medical Observation Period / 中国医学伦理学

To analyze the psychological experience of close contacts with COVID-19 during the centralized isolation medical observation (centralized isolation for short) period, 17 close contacts of COVID-19 who received medical isolation observation at designated locations in a city of China were selected by purposive sampling using qualitative study methods, and the interviewees were interviewed by full-coverage structured telephone. By analyzing the obtained data and summarizing the themes, it was found that the psychological status of the COVID-19 close contacts during the centralized isolation period could be classified into three themes: obvious negative emotions, somatization of psychological problems, and demand for social support. During the epidemic, when close contacts were quarantined as high-risk groups, they often had complex and difficult psychological experiences. Relevant departments should actively carry out online guidance channels and offline knowledge dissemination, actively conduct psychological interventions for close contacts, assist them to develop life plans to enrich the life of the centralized isolation period. At the same time, relevant departments should do a good job in logistics and life support services, create a comfortable isolated living environment, and effectively relieve the negative emotions of close contacts of COVID-19 during centralized isolation through a variety of positive and effective measures to ensure physical and mental health.

Effect of low temperature on the shaping of yeast-derived metabolite compositions during wine fermentation.

Low-temperature fermentation is considered to enrich the aroma of wine. The metabolism of Saccharomyces cerevisiae responding to low temperatures is intricate and this complexity is further enhanced by various strains and culture media. However, the real effects of low-temperature fermentation on yeast metabolism are unclear. Aiming to clarify the yeast-derived metabolite formation in a low-temperature winemaking range, fermentations were performed at 10, 15, and 20 °C, using five wine yeast strains in two media respectively. Tolerance toward low temperatures and metabolite compositions (including basic chemical compositions and volatile aroma compounds) of wine yeasts were analyzed. Results showed that ethanol, ethyl acetate, and ethyl butanoate increased with the temperature decreasing, while acetic acid, phenylethanol, phenylethyl acetate, ethyl decanoate, and ethyl hexadecanoate decreased with decreasing temperature. The linear relationship between fermentation temperature and the formation of ethanol, acetic acid, and phenylethanol might be fundamentally due to the growth changes caused by temperature. The enhanced production of ethyl acetate and ethyl butyrate followed by decreasing temperature probably resulted from low-temperature-stimulated enzymes in metabolic pathways. These findings reveal a typical profile of yeast-derived metabolites at low-temperature fermentation and provide evidence to support the application of low-temperature winemaking in the wine industry.

Programmed genome editing by a miniature CRISPR-Cas12f nuclease.

The RNA-guided CRISPR-associated (Cas) nucleases are versatile tools for genome editing in various organisms. The large sizes of the commonly used Cas9 and Cas12a nucleases restrict their flexibility in therapeutic applications that use the cargo-size-limited adeno-associated virus delivery vehicle. More compact systems would thus offer more therapeutic options and functionality for this field. Here, we report a miniature class 2 type V-F CRISPR-Cas genome-editing system from Acidibacillus sulfuroxidans (AsCas12f1, 422 amino acids). AsCas12f1 is an RNA-guided endonuclease that recognizes 5' T-rich protospacer adjacent motifs and creates staggered double-stranded breaks to target DNA. We show that AsCas12f1 functions as an effective genome-editing tool in both bacteria and human cells using various delivery methods, including plasmid, ribonucleoprotein and adeno-associated virus. The small size of AsCas12f1 offers advantages for cellular delivery, and characterizations of AsCas12f1 may facilitate engineering more compact genome-manipulation technologies.

Improvement of protein production by engineering a novel antiapoptotic baculovirus vector to suppress the expression of Sf-caspase-1 and Tn-caspase-1.

The baculovirus expression vector system (BEVS) is an attractive manufacturing platform for recombinant protein production in insect cells. However, baculovirus infection commonly induces host apoptosis in 3-4 days which would subsequently terminate the protein expression. Previous studies have proved that protein production by BEVS can be elevated in apoptosis-suppressed insect cells. We also developed a baculovirus vector in our previous report to inhibit the apoptosis and improve protein production in Sf9 cells. In this study, we designed five short hairpin RNA (shRNA) expression cassettes targeting a conserved region in Spodoptera frugiperda caspase-1 (Sf-caspase-1) and Trichoplusia ni caspase-1 (Tn-caspase-1), and found that introduction of C to T mutations within the stem region of the expression cassette was beneficial for the heterologous protein expression. One of the improved shRNA expression cassettes was knocked into a bacmid with the deletion of several nonessential genes. The novel baculovirus vector demonstrated the ability to suppress cell apoptosis in both Sf9 and High Five cells, and exhibited superior recombinant protein productivity of intracellularly expressed GFP and firefly luciferase and secreted glycoprotein OD-Fc. The antiapoptotic baculovirus vector developed in this study could serve as a useful tool for the protein production in scientific research and pharmaceutical industries.

Chlorogenic acid inhibits proliferation in human hepatoma cells by suppressing noncanonical NF-κB signaling pathway and triggering mitochondrial apoptosis.

Chlorogenic acid (CGA), a phenylpropanoid derived from Eucommia ulmoides Oliver, has been shown to exhibit potent cytotoxic and anti-proliferative activities against several human cancers. However, the effects of CGA on hepatocellular carcinoma (HCC) and the underlying mechanisms have not been intensively studied. In this study, the CGA treatment effects on the viability of human hepatoma cells were investigated by MTT assay. Our data showed that CGA could dose-dependently inhibit the activity of human hepatoma cells Hep-G2 and Huh-7, but did not affect the activity and growth of normal human hepatocyte QSG-7701. The genes and pathways influenced by CGA treatment were explored by RNA sequencing and bioinformatics analysis, which identified 323 differentially expressed genes (DEGs) involved in multiple pharmacological signaling pathways such as MAPK, NF-κB, apoptosis and TGF-ß signaling pathways. Further analyses by real-time quantitative PCR, Western blot and flow cytometry revealed that CGA effectually suppressed the noncanonical NF-κB signaling pathway, meanwhile it activated the mitochondrial apoptosis of HCC by upregulation of the BH3-only protein Bcl-2 binding component 3 (BBC3). Our findings demonstrated the potential of CGA in suppressing human hepatoma cells and provided a new insight into the anti-cancer mechanism of CGA.

ß-Cyanoalanine Synthase Regulates the Accumulation of ß-ODAP via Interaction with Serine Acetyltransferase in Lathyrus sativus.

ß-N-Oxalyl-l-α,ß-diaminopropionic acid (ß-ODAP), found in Lathyrus sativus at first, causes a neurological disease, lathyrism, when over ingested in an unbalanced diet. Our previous research suggested that ß-ODAP biosynthesis is related to sulfur metabolism. In this study, ß-cyanoalanine synthase (ß-CAS) was confirmed to be responsible for ß-ODAP biosynthesis via in vitro enzymatic analysis. LsCAS was found to be pyridoxal phosphate (PLP)-dependent via spectroscopic analysis and dual functional via enzymatic activity analysis. Generation of a M135T/M235S/S239T triple mutant of LsCAS, which are the key sites to control the ratio of CAS/cysteine synthase (CS) activity, switches reaction chemistry to that of a CS. LsCAS interactions were further screened and verified via Y2H, BiFC and pull-down assay. It was suggested that LsSAT2 interacts and forms a cysteine regulatory complex (CRC) with LsCAS in mitochondria, which improves LsSAT while reduces LsCAS activities to affect ß-ODAP content positively. These results provide new insights into the molecular regulation of ß-ODAP content in L. sativus.

Metabolic acidosis in critically ill patients with cirrhosis: Epidemiology and short-term mortality risk factors.

BACKGROUND/AIMS: Metabolic acidosis is a common complication in patients with cirrhosis at the intensive care units (ICUs) and associated with increased mortality. The aim of our research was to explore the epidemiology and risk factors of metabolic acidosis in critically ill patients with cirrhosis. MATERIALS AND METHODS: A total of 975 patients with cirrhosis were selected into our study, and all participants were followed up for at least 28 days. Cox regression model and machine-learning algorithm were used to identify the importance of different risk factors, respectively. Finally, an improved prognostic model as Model for End-stage Liver Disease and metabolic acidosis (MELD-MA) was developed. RESULTS: Among the 975 patients with liver cirrhosis, 506 had metabolic acidosis, including 257 patients who had decompensated metabolic acidosis at ICU admission. The 28-day mortality was 41% (206/506) in patients with metabolic acidosis. Bilirubin (hazard ratio (HR): 1.023, 95% confidence interval (CI): 1.011-1.036), international normalized ratio (HR: 1.527, 95% CI: 1.332-1.750), pH (HR: 0.173, 95% CI: 0.047-0.640), BE-Lac (HR: 0.907, 95% CI: 0.868-0.948), and BE-Na (HR: 0.923, 95% CI: 0.859-0.991) were considered as independent prognostic parameters for 28-day mortality. MELD-NA had significantly higher discrimination (area under the receiver operating characteristic curve 0.79) than MELD and Child-Pugh score. CONCLUSION: Critically ill patients with cirrhosis have a high mortality rate and poor prognosis because of the high prevalence of metabolic acidosis. Lactic acidosis is the worst prognosis of all types of metabolic acidosis. MELD-MA performs well on the short-term mortality assessment in critically ill patients with cirrhosis and metabolic acidosis.

Coordinated Regulation of Grape Berry Flesh Color by Transcriptional Activators and Repressors.

Yan73 is a teinturier Vitis vinifera variety with red berry flesh, but the molecular mechanisms underlying its flesh coloration remain unclear. We analyzed the flavonoid metabolic and transcriptome profiles of Yan73 berry red and white flesh using HPLC-ESI-MS/MS and RNA-sequencing technologies. Anthocyanins are the main flavonoids responsible for Yan73 berry flesh color, and the coloration is coordinately regulated by the VvMYBA1 transcriptional activator and VvMYBC2-L1 transcriptional repressor. Furthermore, yeast one- and two-hybrid, dual luciferase, and bimolecular fluorescence complementation assays suggested that VvMYBA1 positively regulates Yan73 berry flesh color via interactions with VvWDR1 and the activation of the VvCHI3, VvOMT, and VvGST4 promoters, whereas VvMYBC2-L1 negatively regulates Yan73 berry flesh color, possibly by competing with the R2R3-MYB transcriptional activators for bHLH partners or by repressing VvOMT and VvGST4 expression. Our findings provide new insights into the molecular mechanisms regulating grape flesh color.

Immersion immunization with recombinant baculoviruses displaying cyprinid herpesvirus 2 membrane proteins induced protective immunity in gibel carp.

Cyprinid herpesvirus 2 (CyHV-2) is the causative pathogen of herpesviral haematopoietic necrosis disease, which has caused huge economic losses to aquaculture industry in China. In this study, nine truncated CyHV-2 membrane glycoproteins (ORF25, ORF25C, ORF25D, ORF30, ORF124, ORF131, ORF136, ORF142A, ORF146) and a GFP reporter protein were respectively expressed using baculovirus surface displaying system. Western blot showed that the proteins were successfully packaged in the recombinant virus particles. In baculovirus transduced gibel carp kidney cells, the target proteins were expressed and displayed on the fish cell surface. Healthy gibel carp were immunized by immersion with the recombinant baculoviruses and the fish treated with phosphate-buffered saline (PBS) were served as mock group. The expression of interleukin-11 (IL-11), interferon α (IFNα) and a complement component gene C3 were significantly up-regulated in most experimental groups, and interferon γ (IFNγ) expression in some groups were also induced after immunization. Subsequently, the immunized gibel carp were challenged by intraperitoneal injection of CyHV-2 virus. All the immunized groups exhibited reduced mortality after CyHV-2 challenge. In the groups immunized with baculoviruses displaying and expressing ORF25, ORF25C and ORF146, the relative percentage survival values reached 83.3%, 87.5% and 70.8%, respectively. Our data suggested that baculovirus-displayed ORF25, ORF25C and ORF146 could be potential vaccine candidates for the prevention of CyHV-2 infection in gibel carp.

Smart Camera Aware Crowd Counting via Multiple Task Fractional Stride Deep Learning.

Estimating the number of people in highly clustered crowd scenes is an extremely challenging task on account of serious occlusion and non-uniformity distribution in one crowd image. Traditional works on crowd counting take advantage of different CNN like networks to regress crowd density map, and further predict the count. In contrast, we investigate a simple but valid deep learning model that concentrates on accurately predicting the density map and simultaneously training a density level classifier to relax parameters of the network to prevent dangerous stampede with a smart camera. First, a combination of atrous and fractional stride convolutional neural network (CAFN) is proposed to deliver larger receptive fields and reduce the loss of details during down-sampling by using dilated kernels. Second, the expanded architecture is offered to not only precisely regress the density map, but also classify the density level of the crowd in the meantime (MTCAFN, multiple tasks CAFN for both regression and classification). Third, experimental results demonstrated on four datasets (Shanghai Tech A (MAE = 88.1) and B (MAE = 18.8), WorldExpo'10(average MAE = 8.2), NS UCF_CC_50(MAE = 303.2) prove our proposed method can deliver effective performance.

A viral expression factor behaves as a prion.

Prions are proteins that can fold into multiple conformations some of which are self-propagating. Such prion-forming proteins have been found in animal, plant, fungal and bacterial species, but have not yet been identified in viruses. Here we report that LEF-10, a baculovirus-encoded protein, behaves as a prion. Full-length LEF-10 or its candidate prion-forming domain (cPrD) can functionally replace the PrD of Sup35, a widely studied prion-forming protein from yeast, displaying a [PSI+]-like phenotype. Furthermore, we observe that high multiplicity of infection can induce the conversion of LEF-10 into an aggregated state in virus-infected cells, resulting in the inhibition of viral late gene expression. Our findings extend the knowledge of current prion proteins from cellular organisms to non-cellular life forms and provide evidence to support the hypothesis that prion-forming proteins are a widespread phenomenon in nature.

Non-Invasive Remote Temperature Monitoring Using Microwave-Induced Thermoacoustic Imaging.

Non-invasive temperature monitoring of tissue at depth in real-time is critical to hyperthermia therapies such as high-intensity focused ultrasound. Knowledge of temperature allows for monitoring treatment as well as providing real-time feedback to adjust deposited power in order to maintain safe and effective temperatures. Microwave-induced thermoacoustic (TA) imaging, which combines the conductivity/dielectric contrast of microwave imaging with the resolution of ultrasound, shows potential for estimating temperature non-invasively in real-time by indirectly measuring the temperature dependent parameters from reconstructed images. In this work, we study the temperature dependent behavior of the generated pressure in the TA effect and experimentally demonstrate simultaneous imaging and temperature monitoring using TA imaging. The proof-of-concept experiments demonstrate millimeter spatial resolution while achieving degree-level accuracy.

Correlation between spiritual health and empathy among undergraduate nursing students / 中国实用护理杂志

Objective To investigate the present situation of spiritual health and empathy of undergraduate nursing students, and explore the correlation between them. Methods Convenient sampling was used and 498 senior nursing students were recruited from 2 universities in Shandong Province. They were investigated using the general questionnaire, the Spiritual Health Scale Short Form and Chinese version of the Interpersonal Reactivity Index. Results The total score of spiritual health was (92.87±12.28), the 5 dimensions of the item score from high to low were connection to others (4.46± 0.62), meaning derived from living (4.15 ± 0.69), self-understanding (4.03 ± 0.72), transcendence (3.95 ± 0.74) and religious attachment (2.60±1.08). The total score of Interpersonal Reactivity Index was (57.50± 8.35), the 5 dimensions of the item score from high to low were perspective taking (2.78±0.61), empathy concern (2.72±0.56), fantasy (2.61±0.57), personal distress (2.32±0.78). Nursing students’spiritual health score and empathy was positively correlated (r=0.334,P<0.01). Regression analysis, single-child (B=0.202, P<0.05), exercise habit (B=0.363, P<0.01), religious belief (B=0.381, P<0.01), academic performance (B=0.205, P<0.05), nursing career in the future (B=0.285, P<0.01) were the influencing factors of nursing students′ spiritual health. Conclusions Spiritual health is closely related to empathy; the higher the level of spiritual health, the higher the empathy ability. It suggests that nursing educators can improve nursing students′ empathy by improving the spiritual health of nursing students.

Anticipatory grief and its influencing factors in major caregivers of patients with advanced cancer / 中国实用护理杂志

Objective To explore the status of anticipatory grief in major caregivers of patients with advanced cancer and its influencing factors. Methods There were 329 major caregivers of patients with advanced cancer recruited between October 2017 and May 2018. The basic data questionnaire, Anticipatory Grief Scale and Perceived Social Support Scale were used in the investigation. Results The scores of anticipatory grief and perceived social support were 88.49 ± 16.47 and 60.33 ± 11.58 respectively. The result of multiple linear regression revealed that relationship with patients, perceived social support, gender and religion were associated with anticipatory grief and could explain 54.2％of the total variance. Conclusions Medical workers should provide assistance to the major caregivers of patients with advanced cancer, respect and encourage the appropriate participation of religious activities, encourage caregivers to fully express their feelings, decrease their adverse emotional reactions and improve their bereavement outcome.

Anticipatory grief and its influencing factors in major caregivers of patients with advanced cancer / 中国实用护理杂志

Objective@#To explore the status of anticipatory grief in major caregivers of patients with advanced cancer and its influencing factors.@*Methods@#There were 329 major caregivers of patients with advanced cancer recruited between October 2017 and May 2018. The basic data questionnaire, Anticipatory Grief Scale and Perceived Social Support Scale were used in the investigation.@*Results@#The scores of anticipatory grief and perceived social support were 88.49±16.47 and 60.33±11.58 respectively. The result of multiple linear regression revealed that relationship with patients, perceived social support, gender and religion were associated with anticipatory grief and could explain 54.2% of the total variance.@*Conclusions@#Medical workers should provide assistance to the major caregivers of patients with advanced cancer, respect and encourage the appropriate participation of religious activities, encourage caregivers to fully express their feelings, decrease their adverse emotional reactions and improve their bereavement outcome.

Correlation between spiritual health and empathy among undergraduate nursing students / 中国实用护理杂志

Objective@#To investigate the present situation of spiritual health and empathy of undergraduate nursing students, and explore the correlation between them.@*Methods@#Convenient sampling was used and 498 senior nursing students were recruited from 2 universities in Shandong Province. They were investigated using the general questionnaire, the Spiritual Health Scale Short Form and Chinese version of the Interpersonal Reactivity Index.@*Results@#The total score of spiritual health was (92.87±12.28), the 5 dimensions of the item score from high to low were connection to others (4.46±0.62), meaning derived from living (4.15±0.69), self-understanding (4.03±0.72), transcendence (3.95±0.74) and religious attachment (2.60±1.08). The total score of Interpersonal Reactivity Index was (57.50±8.35), the 5 dimensions of the item score from high to low were perspective taking (2.78±0.61), empathy concern (2.72±0.56), fantasy (2.61±0.57), personal distress (2.32±0.78). Nursing students’ spiritual health score and empathy was positively correlated (r=0.334, P<0.01). Regression analysis, single-child (B=0.202, P<0.05), exercise habit (B=0.363, P<0.01), religious belief (B=0.381, P<0.01), academic performance (B=0.205, P<0.05), nursing career in the future (B=0.285, P<0.01) were the influencing factors of nursing students′ spiritual health.@*Conclusions@#Spiritual health is closely related to empathy; the higher the level of spiritual health, the higher the empathy ability. It suggests that nursing educators can improve nursing students′ empathy by improving the spiritual health of nursing students.

A follow-up study about self-acceptance and automatic thoughts in permanent colostomy patients / 中国实用护理杂志

Objective To explore the dynamic changes and correlation of self-acceptance and automatic thoughts in permanent colostomy patients. Methods There were 89 low position rectal cancer patients after Miles and permanent colostomy were recruited from the Department of Oncological Surgery of the First Affiliated Hospital of Xi'an Jiaotong UniversityfromFebruary 2016 to October 2016. Self-acceptance Questionnaire and Automatic Thoughts Questionnaire were used to collect data in 1 month, 3 months, 6 months and 12 months after colostomy. Results The self-acceptance scores were 33.64 ± 5.52, 33.24 ± 5.71, 34.56 ± 6.04 and 38.91 ± 6.33 respectively, which had significant differences (F=11.012, P<0.01). The automatic thoughts scores were 63.20 ± 16.78, 60.88 ± 16.53, 58.64 ± 15.87 and 55.32 ± 14.76 respectively, which had significant differences (F=3.462, P<0.05). The correlation between self-acceptance and automatic thoughts were-0.576--0.372, which were all significant (P<0.01). Conclusion There were negative correlations between self-acceptance and automatic thoughts in permanent colostomy patients within 1 year after operation. Health professionals should learn to identify patients′automatic thoughts accurately, lead off negative emotions, promote early self-acceptance and improve their quality of life.