Reverse genetics systems for SARS-CoV-2: Development and applications.

The recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused serious harm to human health and struck a blow to global economic development. Research on SARS-CoV-2 has greatly benefited from the use of reverse genetics systems, which have been established to artificially manipulate the viral genome, generating recombinant and reporter infectious viruses or biosafety level 2 (BSL-2)-adapted non-infectious replicons with desired modifications. These tools have been instrumental in studying the molecular biological characteristics of the virus, investigating antiviral therapeutics, and facilitating the development of attenuated vaccine candidates. Here, we review the construction strategies, development, and applications of reverse genetics systems for SARS-CoV-2, which may be applied to other CoVs as well.

Pectate Lyase from Fusarium sacchari Induces Plant Immune Responses and Contributes to Virulence.

Fusarium sacchari is one of the primary pathogens causing Pokkah Boeng disease (PBD) in sugarcane in China. Pectate lyases (PL), which play a critical role in pectin degradation and fungal virulence, have been extensively studied in major bacterial and fungal pathogens of a wide range of plant species. However, only a few PLs have been functionally investigated. In this study, we analyzed the function of the pectate lyase gene, FsPL, from F. sacchari. FsPL is a key virulence factor of F. sacchari and can induce plant cell death. FsPL also triggers the pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) response in Nicotiana benthamiana, as reflected by increases in reactive oxygen species (ROS) production, electrolyte leakage, and callose accumulation, as well as the upregulation of defense response genes. In addition, our study also found that the signal peptide of FsPL was necessary for induced cell death and PTI responses. Virus-induced gene silencing showed that FsPL-induced cell death in Nicotiana benthamiana was mediated by leucine-rich repeat (LRR) receptor-like kinases BAK1 and SOBIR1. Thus, FsPL may not only be a critical virulence factor for F. sacchari but may also induce plant defense responses. These findings provide new insights into the functions of pectate lyase in host-pathogen interactions. IMPORTANCE Pokkah Boeng disease (PBD) is one of the main diseases affecting sugarcane in China, seriously damaging sugarcane production and economic development. Therefore, it is important to clarify the pathogenic mechanisms of this disease and to provide a theoretical basis for the breeding of PBD-resistant sugarcane strains. The present study aimed to analyze the function of FsPL, a recently identified pectate lyase gene from F. sacchari. FsPL is a key virulence factor of F. sacchari that induces plant cell death. Our results provide new insights into the function of pectate lyase in host-pathogen interactions.

Intelligent Power System Stability Assessment and Dominant Instability Mode Identification Using Integrated Active Deep Learning.

Simulation analysis is critical for identifying possible hazards and ensuring secure operation of power systems. In practice, large-disturbance rotor angle stability and voltage stability are two frequently intertwined stability problems. Accurately identifying the dominant instability mode (DIM) between them is important for directing power system emergency control action formulation. However, DIM identification has always relied on human expertise. This article proposes an intelligent DIM identification framework that can discriminate among stable status, rotor angle instability, and voltage instability based on active deep learning (ADL). To reduce human expert efforts required to label the DIM dataset when building DL models, a two-stage batch-mode integrated ADL query strategy (preselection and clustering) is designed for the framework. It samples only the most helpful samples to label in each iteration and considers both information contents and diversity in them to improve query efficiency, significantly reducing the required number of labeled samples. Case studies conducted on a benchmark power system (China Electric Power Research Institute (CEPRI) 36-bus system) and a practical large-area power system (Northeast China Power System) reveal that the proposed approach outperforms conventional methods in terms of accuracy, label efficiency, scalability, and adaptability to operational variability.

[Randomized controlled trial outcome indicators of traditional Chinese medicine for gastrointestinal dysfunction in sepsis in recent two years].

The purpose of the study is to analyze the outcomes of randomized controlled trial(RCT) of Chinese herbal medicine formula(CHMF) in the treatment of gastrointestinal dysfunction in sepsis in recent two years. We systematically searched four Chinese databases, three English databases, and two clinical trial registries to analyze the reports of outcome indicators of clinical trials, and evaluated the risk of bias by using the ROB tool of Cochrane Collaboration. After screening, 55 clinical RCTs were included. The results showed that the current clinical studies of gastrointestinal dysfunction in sepsis reported the efficacy and safety indicators. The efficacy indicators included APACHE Ⅱ scores, gastrointestinal dysfunction scores, bowel sound scores, and inflammatory indicator such as C-reactive protein and procalcitonin. The safety indicators mainly include gastrointestinal reactions, skin reactions, and other adverse events and adverse reactions. However, there was no distinction between primary and secondary outcomes. The relevant indicators of health economics were not reported, and the quality of research methodology was poor. Therefore, we suggest that future researchers should be well prepared in the top-level design stage and actively construct the core outcome set, so as to improve the quality of clinical trials.

Developing and Validating Risk Scores for Predicting Major Cardiovascular Events Using Population Surveys Linked with Electronic Health Insurance Records.

A risk prediction model for major cardiovascular events was developed using population survey data linked to National Health Insurance (NHI) claim data and the death registry. Another set of population survey data were used to validate the model. The model was built using the Nutrition and Health Survey in Taiwan (NAHSIT) collected from 1993-1996 and linked with 10 years of events from NHI data. Major adverse cardiovascular events (MACEs) were identified based on hospital admission or death from coronary heart disease or stroke. The Taiwanese Survey on Hypertension, Hyperglycemia, and Hyperlipidemia (TwSHHH), conducted in 2002 was used for external validation. The NAHSIT data consisted of 1658 men and 1652 women aged 35-70 years. The incidence rates for MACE per 1000 person-years were 13.77 for men and 7.76 for women. These incidence rates for the TwSHHH were 7.27 for men and 3.58 for women. The model had reasonable discrimination (C-indexes: 0.76 for men; 0.75 for women), thus can be used to predict MACE risks in the general population. NHI data can be used to identify disease statuses if the definition and algorithm are clearly defined. Precise preventive health services in Taiwan can be based on this model.

Assessment of the anti-rheumatoid arthritis activity of Gastrodia elata (tian-ma) and Radix aconitic lateralis preparata (fu-zi) via network pharmacology and untargeted metabolomics analyses.

AIM: Gastrodia elata and Radix aconiti lateralis preparrata are respectively named as Tian-Ma and Fu-Zi (TF) in Chinese. We explored the active components against rheumatoid arthritis (RA) from an extensively used couplet of Chinese herbs, Gastrodia elata and Radix aconiti lateralis preparata (TF) via untargeted metabolomics and network pharmacological approaches. METHODS: Water extracts of TF were mixed at ratios 1:1, 3:2 and 2:3 (w/w). Ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) was then utilized as metabolomics screening. Human Metabolome (http://www.hmdb.ca/) and Lipidmaps (http://www.lipidmaps.org/) databases were used to annotate detected compounds. Further identification of vital genes and important pathways associated with the anti-RA properties of the TF preparations was done via network pharmacology, and verified by real-time quantitative polymerase chain reaction (RT-qPCR). RESULTS: Four key compounds involved in unsaturated fatty acid biosynthesis and isoflavonoid biosynthesis were identified through metabolomics analyses. Three key components of TF associated with anti-RA activity were linoleic acid, daidzein, and daidzin. Results of RT-qPCR revealed that all 3 tested TF couplets (1:1, 3:2, and 2:3) markedly suppressed the transcription of PTGS2. These results were consistent with our network pharmacological predictions. CONCLUSIONS: The anti-RA properties of Tian-Ma and Fu-Zi are associated with the inhibition of arachidonic acid metabolism pathway.

Hemostatic potential of cold-stored non-leukoreduced whole blood over time: An assessment of platelet function and thrombin generation for optimal shelf life.

OBJECTIVES: Cold-stored low-titer whole blood (WB) is becoming increasingly used as the preferred product for initial hemorrhagic shock resuscitation. The purpose of this study was to identify whether the current 21-day shelf life is the optimal duration for storage of WB, maintaining hemostatic efficacy. METHODS: Five units of fresh low-titer group O WB (non-leukoreduced) were acquired from our regional blood center. These units were stored at 4°C for up to 21 days as per current clinical storage guidelines in our emergency department. Hemostatic parameters were measured in vitro at 0 days, 7 days, 14 days, and 21 days. Assessments of hemostatic potential included cell count, rapid thrombelastography (r-TEG) and kaolin thrombelastography (TEG), multiplate impedance aggregometry, and calibrated automated thrombogram (CAT). Univariate analysis, including one-way analysis of variance with repeated measures, was performed (STATA 12.1). RESULTS: Compared with baseline product (0 days), both platelet count and platelet function of WB showed sharp decreases at 7 days and again at 14 days. Platelet function deterioration was noted by r-TEG c (MA), TEG-MA, and multiplate arachidonic acid and adenosine diphosphate (all p < 0.001). With respect to clot initiation, r-TEG ACT and TEG R-time were similar over the 21-day shelf life (p = 0.058 and p = 0.620, respectively). Thrombin generation assessed by CAT demonstrated stable endogenous thrombin potential over the course of storage (p = 0.162), but increased peak thrombin generation and quicker time to peak generation after 7 days. CONCLUSION: While the platelet function of WB degrades significantly at 7 days (and again at 14 days), clot initiation remains stable over time, and thrombin generation appears to be improved at 7 days. This study supports a current storage limit for cold-stored, low-titer WB of 14 days.

Cost-drivers of medical expenses in burn care management.

BACKGROUND: Profound differences exist in the cost of burn care globally, thus we aim to investigate the affected factors and to delineate a strategy to improve the cost-effectiveness of burn management. METHODS: A retrospective analysis of 66 patients suffering from acute burns was conducted from 2013 to 2015. The average age was 26.7 years old and TBSA was 42.1% (±25.9%). We compared the relationship between cost and clinical characteristics. RESULTS: The estimated cost of acute burn care with the following formula (10,000 TWD) = -19.80 + (2.67 × percentage of TBSA) + (124.29 × status of inhalation injury) + (147.63 × status of bacteremia) + (130.32 × status of respiratory tract infection). CONCLUSION: The majority of the cost were associated with the use of antibiotics and burns care. Consequently, it is crucial to prevent nosocomial infection in order to promote healthcare quality and reduce in-hospital costs.

The thermodynamics of guest complexation to octa-acid and tetra-endo-methyl octa-acid: reference data for the sixth statistical assessment of modeling of proteins and ligands (SAMPL6).

Although computer-aided drug design has greatly improved over time, its application in the pharmaceutical industry is still limited by the accuracy of association constant predictions. Towards improving this situation, the Statistical Assessment of the Modeling of Proteins and Ligands (SAMPL) is a series of community-wide blind challenges aimed to advance computational techniques as standard predictive tools in rational drug design (https://en.wikipedia.org/wiki/SAMPL_Challenge). As an empirical contribution to the sixth assessment (SAMPL6), we report here the association constant (Ka ) and thermodynamic parameters (∆G, ∆H, -T∆S) of eight guests (G0-G7) binding to two subtly different hosts (OA and TEMOA) using ITC. Both hosts contain a unique, well-defined binding pocket capable of storing guests with up to ten non-hydrogen atoms, whilst the selection of amphiphilic guests contain a range of saturated and unsaturated substituents from C6 to C10. The thermodynamic data from this study will allow the challenge participants of SAMPL6 to test the accuracy of their computational protocols for calculating host-guest affinities.

A Spatio-Temporal Pattern and Socio-Economic Factors Analysis of Improved Sanitation in China, 2006⁻2015.

Ensuring an adequate and safe access to sanitation is essential to prevent diseases. Using provincial spatial panel data reported in the China Health Statistical Yearbook and the China Statistical Yearbook, this paper analyzed the spatio-temporal characteristics of improved rural sanitation in 30 Chinese provinces during the period 2006⁻2015, and analyzed factors that may affect improved sanitation rates in rural China. Spatial autocorrelations of improved sanitation rates were computed via Global and Local Moran's I firstly, and then, inter-provincial disparities of improved sanitation were assessed by using the Theil index estimator; finally, the spatial panel model was employed to examine the potential socio-economic factors. Spatial autocorrelations results suggested that the provincial improved sanitation rates changes affect both the provinces themselves and the adjacent regions; Analysis of the spatial panel model revealed that factors such as GDP per capita, investment proportion ratio, centralized water supply, rural residents' expenditure were positively associated with improved sanitation rates, and illiteracy rate of people older than 15 was negatively related with improved sanitation rates. Socio-economic factors had affected the improved sanitation rates in 30 provinces in rural China. Thus, a series of policies, socio-economic measures and personal latrine literacy education should be given to improve the status of improved sanitation rates in rural China.

The significance of magnetic resonance imaging in severe femoral trochlear dysplasia assessment.

The clinical diagnosis of femoral trochlear dysplasia primarily relies on imaging. In the past, plain imaging was the major source of diagnosis. The present study investigated the application of magnetic resonance imaging (MRI) in the objective assessment of severe femoral trochlear dysplasia. A retrospective analysis was performed on knee MRIs from 30 normal subjects (30 knees) and 59 patients (61 knees) with severe femoral trochlear dysplasia based on the Dejour morphological classification. Cartilage and subchondral bone landmarks were used to compare a series of measurements between patient and control groups. These measurements included the femoral trochlear groove depth, sulcus angle, the lateral trochlear inclination, trochlear facet asymmetry, the femoral medial and lateral condyle symmetry, and the ratios between the femoral medial/lateral condyles and the maximal trochlear width. The measurement values based on the two types of landmarks were also compared within the patient and control groups, separately. In addition, the femoral trochlear groove depth, sulcus angle, lateral trochlear inclination and trochlear facet asymmetry of patients with different Dejour types were compared. Significant differences were observed in the femoral trochlear groove depth, sulcus angle, lateral trochlear inclination and trochlear facet asymmetry between the patient and control groups (P<0.05). Based on the two types of landmark, all indexes were significantly different (P<0.05; with the exception of lateral trochlear inclination) between the patient and control groups. Among patients with various Dejour types, the femoral trochlear groove depth, sulcus angle, lateral trochlear inclination and trochlear facet asymmetry demonstrated no significant differences. MRI exhibited advantages in revealing articular cartilage over conventional radiography and computed tomography. Therefore, cartilage landmarks in MRI images may be utilized to objectively evaluate femoral trochlear dysplasia in patients with severe femoral trochlear dysplasia.

Evaluation of two observational pain assessment scales during the anaesthesia recovery period in Chinese surgical older adults.

AIMS AND OBJECTIVES: To evaluate the reliability and validity of the Pain Assessment in Advanced Dementia scale and the Checklist of Nonverbal Pain Indicators in Chinese older adults post surgery during the anaesthesia recovery period. BACKGROUND: Pain assessment in older surgical patients is complicated by factors such as anaesthesia and opioid administration. Although observational pain behavioural assessment tools have been validated for those unable to self-report, research on their application during the anaesthesia recovery period is limited. DESIGN: A prospective correlational design. METHODS: Ninety-three older patients admitted for scheduled abdominal surgery were recruited in a university-affiliated hospital. The two observational scales were used to conduct pain assessments during the anaesthesia recovery period. On the first and the third postoperative day, participants recalled their pain intensity during the recovery period using the Numeric Rating Scale or the Faces Pain Scale-Revised. RESULTS: The internal consistency reliability of the Pain Assessment in Advanced Dementia scale and the Checklist of Nonverbal Pain Indicators was 0·81 and 0·69 respectively. The correlation between scores of the two observational scales was 0·95. The recalled self-reports of pain intensity were significantly correlated. The correlation of the Pain Assessment in Advanced Dementia scale and patients' recalled self-reports was 0·55, 0·54, and the correlation between the Checklist of Nonverbal Pain Indicators and the two recalled pain scores was both 0·60. CONCLUSIONS: Both the two observational scales had good reliability and validity when used to assess pain in Chinese surgical older adults during the anaesthesia recovery period. RELEVANCE TO CLINICAL PRACTICE: Observational pain scales can be useful as a tool for patients unable to self-report. Accurate use of one of the observational pain tools can help identify pain during the anaesthesia recovery period, when patients are unable to self-report, to support effective pain management during this period.

Assessment of the antibacterial activity and the antidiarrheal function of flavonoids from bayberry fruit.

Chinese bayberry fruits are used as a folk medicine to cure diarrhea. However, the active compounds have not yet been reported. We found that bayberry fruit extract showed significant antibacterial activity against Salmonella, Listeria, and Shigella, and the minimal inhibitory concentration (MIC) ranged from 2.07 to 8.28 mg/mL. Positive relationships were found between the antibacterial activity and the total polyphenol (r = 0.88) and flavonoid contents (r = 0.92) of samples using different extraction times. The active compounds showed green or blue fluorescence under UV light using the bioautography method and were purified using a polyamide column. The fraction F1 with the most activity was comprised of flavonoids, which included cyanidin-3-O-glucoside, myricetin deoxyhexoside, quercetin-3-O-glucoside, and quercetin deoxyhexoside, and it also possessed an antidiarrheal activity (p < 0.10) at 80 mg/kg in mice. These findings provide scientific evidence for the antidiarrheal function of bayberry.

Assessment of coronary heart disease risk by combined analysis of coagulation factors.

OBJECTIVE: Prospective studies have reported a positive association of coagulation factors with risk of coronary heart disease (CHD). It is unclear whether these coagulation factors interact. METHODS AND RESULTS: Using a prospective case-cohort design, we analyzed by Cox proportional hazard regression interactions between soluble thrombomodulin (sTM) and fibrinogen, factor VIII (FVIII), FVII, or plasminogen activator inhibitor-1 (PAI-1) in 410 CHD cases and 721 non-cases from the Atherosclerosis Risk in Communities (ARIC). There was a significant interaction between sTM and fibrinogen (p=0.027). We next assessed risk ratios (RR) by combined tertile analysis. Combined analysis revealed that being in the upper sTM tertile counteracted the CHD risk imposed by higher fibrinogen whereas being in the lower sTM tertile amplified the CHD risk of higher fibrinogen. sTM and fibrinogen mutually influenced CHD incidence in a concentration-dependent manner. When analyzed as single factors by tertiles, FVIII, FVII and PAI-1 were not associated with CHD. However, when analyzed together with sTM, FVIII and PAI-1 were both positively associated with CHD for those in the lower sTM tertile. CONCLUSION: There is a complex interaction between sTM and prothrombotic coagulation factors. Combined analysis improves CHD risk assessment.

Role of sonography in the preoperative assessment of neurilemmoma.

This case report illustrates the role of high-resolution sonography in the preoperative assessment of a neurilemmoma of the median nerve in the forearm. Sonography identified the 3-dimensional localization of the nerve tumor its origin from a nerve fascicle, and its relationship with noninvolved nerve fascicles, facilitating the surgeon's approach to preserve noninvolved fascicles.

Quantitative assessment of osteoporosis from the tibia shaft by ultrasound techniques.

Bone mineral density (BMD) is used as a clinical estimate of the risk of fracture. Ultrasound provides an alternative or complement to X-ray based methods of bone densitometry for determining BMD. Among ultrasonic characteristics, the speed of sound (SOS) is a useful tool for assessment of osteoporosis because, as recently reported, it represents a combination of density and compressibility of bone tissue. Thus, it might provide better information on bone quality to estimate the fracture risk. In this paper, a dual-transducer ultrasound technique was employed to measure the mean ultrasound propagation speed of the cortical layer as well as the cancellous layer at the tibia shaft. Encouraging results from 18 outpatients showed a high correlation (r = 0.93) between measurements of BMD and those from dual energy X-ray absorptiometry (DEXA).