Generation of Bioactivity-Tailored FK506/FK520 Analogs by CRISPR Editing in Streptomyces tsukubaensis.

For a potential application of FK506 in the treatment of acute kidney failure only the FKBP12 binding capability of the compound is required, while the immunosuppressive activity via calcineurin binding is considered as a likely risk to the patients. The methoxy groups at C13 and C15 are thought to have significant influence on the immunosuppressive activity of the molecule. Consequently, FK506 analogs with different functionalities at C13 and C15 were generated by targeted CRISPR editing of the AT domains in module 7 and 8 of the biosynthetic assembly line in Streptomyces tsukubaensis. In addition, the corresponding FK520 (C21 ethyl derivative of FK506) analogs could be obtained by media adjustments. The compounds were tested for their bioactivity in regards to FKBP12 binding, BMP potentiation and calcineurin sparing. 15-desmethoxy FK506 was superior to the other tested analogs as it did not inhibit calcineurin but retained high potency towards FKBP12 binding and BMP potentiation.

Effects of Postural Resonance on Skin Sympathetic Nerve Activity and Blood Pressure: A Pilot Study Evaluating Vascular Tone Baroreflex Stimulation Through Biofeedback.

Heart rate and vascular tension baroreflex exhibit resonance characteristics at approximately 0.1 and 0.03 Hz. In this study, we aimed to induce postural resonance (PR) through rhythmic postural adjustments. To assess the viability of this technique, we investigated the acute impacts of postural resonance on blood pressure, sympathetic nerve activity, and mood. Fifteen healthy study participants, consisting of 8 males and 7 females, were selected for this self-controlled study. Skin sympathetic nerve activity was continuously monitored during both the intervention and stress test on the experimental day. After PR intervention, the diastolic blood pressure and mean arterial pressure in the PR group exhibited significant reductions compared to the CON group (P = 0.032, CON = 71.67 ± 2.348, PR = 64.08 ± 2.35; P = 0.041, CON = 75.00 ± 2.17, PR = 81.67 ± 2.17). After PR intervention both left brachial ankle pulse wave velocity and right brachial ankle pulse wave velocity exhibited a significant reduction compared to pre-intervention levels (from 1115.86 ± 150.08 to 1048.43 ± 127.40 cm/s, p < 0.001; 1103.86 ± 144.35 to 1060.43 ± 121.35 cm/s, p = 0.018). PR intervention also led to a significant decrease in burst frequency and duration (P = 0.049; CON = 8.96 ± 1.17, PR = 5.51 ± 1.17) and a noteworthy decrease in burst amplitude and burst threshold during the cold-pressor test (P = 0.002; P = 0.002). Additionally, VAS scores exhibited a substantial increase following PR (P = 0.035, CON = 28.4 ± 4.49, PR = 42.17 ± 4.10). PR can induce resonance effects within the cardiovascular system, resulting in the effective reduction of blood pressure, skin sympathetic nerve activity and pulse wave velocity, and decreased burst amplitude and burst threshold of the sympathetic nerve during the cold-pressor test.

Long non-coding RNA LOC103222771 promotes infection of porcine reproductive and respiratory syndrome virus in Marc-145 cells by downregulating Claudin-4.

Porcine reproductive and respiratory syndrome (PRRS) is an important swine disease caused by infection of porcine reproductive and respiratory syndrome virus (PRRSV), which leads to huge loss in swine industry. How to effectively control PRRS is challenging. Long non-coding RNA (lncRNA) are key regulator of viral infections and anti-virus immunological responses, therefore, further understanding of lncRNAs will aid to identification of novel regulators of viral infections and better design of prevention and control strategies to viral infection related diseases and immune disorders. We demonstrated that PRRSV infection upregulated the expression of lncRNA LOC103222771 in Marc-145 cells and porcine alveolar macrophage cells (PAMs) and that LOC103222771 is mainly located in cytoplasm. Knockdown of LOC103222771 could inhibit the PRRSV infection in Marc-145 cells. RNA-seq analysis and subsequent validation revealed increased expression of Claudin-4 (CLDN4) in Marc-145 when LOC103222771 was specifically downregulated，suggesting that LOC103222771 might be an upstream regulator of CLDN4, an important component of tight junctions for establishment of the paracellular barrier that controls the flow of molecules in the intercellular space between epithelial cells. We and others showed that Downregulation of CLDN4 could boost the infection of PRRSV. Collectively, LOC103222771/CLDN4 signal axis might be a novel mechanism of PRRSV pathogenesis, implying a potential therapeutic target against PRRSV infection.

Association between composite dietary antioxidant index and handgrip strength in American adults: Data from National Health and Nutrition Examination Survey (NHANES, 2011-2014).

Background: The Composite Dietary Antioxidant Index (CDAI), a composite score of multiple dietary antioxidants (including vitamin A, C, and E, selenium, zinc, and carotenoids), represents an individual's comprehensive dietary antioxidant intake profile. CDAI was developed based on its combined effect on pro-inflammatory markers Tumor Necrosis Factor-α (TNF-α) and anti-inflammatory effects of Interleukin-1ß (IL-1ß), which are associated with many health outcomes, including depression, all-cause mortality, colorectal cancer, etc. Handgrip strength is used as a simple measure of muscle strength, not only is it highly correlated with overall muscle strength, but also serves as a diagnostic tool for many adverse health outcomes, including sarcopenia and frailty syndromes. Purpose: The association between CDAI and Handgrip strength (HGS) is currently unclear. This study investigated the association between CDAI (including its components) and HGS in 6,019 American adults. Method: The research data were selected from the 2011-2014 National Health and Nutrition Survey (NHANES), and a total of 6,019 American adults were screened and included. A weighted generalized linear regression model was used to evaluate CDAI (including its components) and HGS. Results: (1) CDAI was significantly positively correlated with HGS (ß = 0.009, 0.005∼0.013, P < 0.001), and the trend test showed that compared with the lowest quartile of CDAI, the highest quartile of CDAI was positively correlated with HGS (ß = 0.084, 0.042∼0.126, P = 0.002) and significant in trend test (P for trend < 0.0100). Gender subgroup analysis showed that male CDAI was significantly positively correlated with HGS (ß = 0.015, 0.007∼0.023, P = 0.002), and the trend test showed that compared with the lowest quartile of CDAI, the highest quartile of CDAI was positively correlated with HGS (ß = 0.131, 0.049∼0.213, P = 0.006) and the trend test was significant (P for trend < 0.0100). There was no correlation between female CDAI and HGS, and the trend test was not statistically significant (P > 0.05). (2) The intake of dietary vitamin E, Zinc and Selenium showed a significant positive correlation with HGS (ß = 0.004, 0.002∼0.007, P = 0.006; ß = 0.007, 0.004∼0.009, P < 0.001; ß = 0.001, 0.001∼0.001, P < 0.001), vitamin A, vitamin C and carotenoid were significantly associated with HGS in the Crude Model, but this significant association disappeared in the complete model with the increase of control variables. Gender subgroup analysis showed that in model 3, male dietary intake levels of vitamin E, Zinc, and Selenium were significantly positively correlated with HGS (ß = 0.005, 0.002∼0.009, P = 0.011; ß = 0.007, 0.004∼0.011, P = 0.001; ß = 0.001, 0.001∼0.001, P = 0.004), the rest of the indicators had no significant correlation with HGS. Among the female subjects, dietary zinc intake was significantly positively correlated with HGS (ß = 0.005, 0.001∼0.008, P = 0.008), and there was no significant correlation between other indicators and HGS (P > 0.05). Conclusion: There was an association between the CDAI and HGS, but there was a gender difference, and there was an association between the CDAI and HGS in male, but the association was not significant in female. Intake of the dietary antioxidants vitamin E, selenium, and zinc was associated with HGS in male, but only zinc was associated with HGS among dietary antioxidants in female.

TIPE2 acts as a tumor suppressor and correlates with tumor microenvironment immunity in epithelial ovarian cancer.

BACKGROUND: Epithelial ovarian cancer (EOC) is one of the deadliest gynecologic cancers. The etiology of EOC has still not been elucidated thoroughly. Tumor necrosis factor-α-induced protein 8-like2 (TNFAIP8L2, TIPE2), an important regulator of inflammation and immune homeostasis, plays a critical role in the progression of various cancers. This study aims to investigate the role of TIPE2 in EOC. METHODS: Expression of TIPE2 protein and mRNA in EOC tissues and cell lines was examined using Western blot and quantitative real-time PCR (qRT-PCR). The functions of TIPE2 in EOC were investigated by cell proliferation assay, colony assay, transwell assay, and apoptosis analysis in vitro. To further investigate the regulatory mechanisms of TIPE2 in EOC, RNA-seq and western blot were performed. Finally, the CIBERSORT algorithm and databases including Tumor Immune Single-cell Hub (TISCH), Tumor Immune Estimation Resource (TIMER), Tumor-Immune System Interaction (TISIDB), and The Gene Expression Profiling Interactive Analysis (GEPIA) were used to elucidate its potential role in regulating tumor immune infiltration in the tumor microenvironment (TME). RESULTS: TIPE2 expression was shown to be considerably lower in both EOC samples and cell lines. Overexpression of TIPE2 suppressed EOC cell proliferation, colony formation, and motility in vitro. Mechanistically, TIPE2 suppressed EOC by blocking the PI3K/Akt signaling pathway, according to bioinformatics analysis and western blot in TIPE2 overexpression EOC cell lines, and the anti-oncogenic potentials of TIPE2 in EOC cells could be partially abrogated by the PI3K agonist, 740Y-P. Finally, TIPE2 expression was positively associated with various immune cells and possibly involved in the regulation of macrophage polarization in ovarian cancer. CONCLUSIONS: We detail the regulatory mechanism of TIPE2 in EOC carcinogenesis, as well as how it correlates with immune infiltration, emphasizing its potential as a therapeutic target in ovarian cancer.

A dual-modal electrochemical aptasensor based on intelligent DNA Walker with cascade signal amplification powered by Nb.BbvCI for Pb2.

As a unique nanomachine, DNA Walker can move continuously along a specific orbit to amplify signal. Therefore, based on DNA Walker and endonuclease assisted signal amplification strategy, a novel dual-mode visual electrochemical aptasensor was constructed for the detection of Pb2+. Ceric dioxide@mesoporous carbon (CeO2/CS)@AuNPs not only could improve the conductivity of sensing interface but also could fix the aptamer. DNA Walker moved on the surface of the electrode to realize the pairing with the Ag-γFe2O3/cDNA probe, forming a special base sequence that could be spliced by the Nb.BbvCI. Under the action of endonuclease Nb.BbvCI, the Ag-γFe2O3/cDNA probe was continuously sheared and the amount on the electrode was decreased to amplify the signal. Besides, the nanoenzyme of Ag-γFe2O3 could catalyze 3'3'5'5'-tetramethylbenzidine (TMB) to blue color realizing the visual detection of Pb2+. The sensor has been successfully applied to the visual and accurate rapid detection of Pb2+ in aquatic products. The fabricated method of the sensor open up a new way for visual and accurate the detection of environmental pollutants.

Association between dynamic cardiorespiratory fitness and aerobic endurance among university students / 中国学校卫生

Objective@#To screen dynamic cardiorespiratory fitness indices for effective assessment of aerobic endurance using incremental load exercise tests.@*Methods@#From March to November 2019, 266 volunteers who were able to understand the trial and voluntarily cooperated with the entire testing process were randomly recruited from 5 universities in Beijing. Gas metabolism and cardiac function were monitored in real time using the German Cortex Metalyzer 3B system and the US Cheetah NICOM system, and maximum oxygen uptake was measured by a linear incremental loading scheme. Aerobic endurance and dynamic cardiopulmonary function indices were selected according to the study design, and the relationship between the two indicators was analyzed using correlation and typicality correlation.@*Results@#The absolute maximum oxygen uptake (VO 2max ), relative maximum oxygen uptake, maximum stroke volume (SV max ), and maximum stroke volume index (SVI max ) were significantly higher in males than in females, and the dynamic cardiopulmonary function index K was significantly lower than in females ( t =17.8, 10.1, 8.5, 4.3 , -6.3, P < 0.05 ). Simple correlation revealed that absolute VO 2max and relative VO 2max were negatively correlated with K and b, and absolute VO 2max and relative VO 2max were positively correlated with SV max and SVI max ( P <0.01); Both dynamic cardiopulmonary function indices were significantly correlated with aerobic endurance. Aerobic endurance indexes closely related to dynamic cardiopulmonary function were absolute VO 2max , relative VO 2max ; dynamic cardiopulmonary function indexes closely related to aerobic endurance were K, b, SV max , SVI max .@*Conclusion@#Improvement of dynamic cardiorespiratory fitness contributes to the improvement of aerobic endurance, the dynamic cardiorespiratory fitness indices K, b, SV max , and SVI max can be used as important candidates to predict the assessment of aerobic exercise capacity.

Effects of trunk posture on cardiovascular and autonomic nervous systems: A pilot study.

Objective: Although regular and moderate physical activity has been shown to improve the cardiovascular and autonomic nervous systems, little has been done to study the effects of postural changes in the movement on the heart and autonomic nervous system. To uncover changes in cardiac function and autonomic nerves induced by different underlying posture transitions and explore which trunk postures lead to chronic sympathetic activation. Therefore, this study investigated the effects of trunk posture on the cardiovascular and autonomic nervous systems. Methods: Twelve male subjects (age 24.7 ± 1.3) underwent this study. The non-invasive cardiac output NICOM monitoring equipment and the FIRSTBEAT system are used to dynamically monitor seven trunk postures in the sitting position simultaneously (neutral position, posterior extension, forward flexion, left lateral flexion, right lateral flexion, left rotation, right rotation). Each posture was maintained for 3 min, and the interval between each movement was 3 min to ensure that each index returned to the baseline level. Repeated analysis of variance test was used to compare and analyze the differences in human cardiac function, heart rate variability index, and respiratory rate under different postures. Results: Compared with the related indicators of cardiac output in a neutral trunk position: the cardiac index (CI) was significantly reduced in forwarding flexion and left rotation (3.48 ± 0.34 vs. 3.21 ± 0.50; 3.48 ± 0.34 vs. 3.21 ± 0.46, Δ L/(min/m2)) (p = 0.016, p = 0.013), cardiac output decreased significantly (6.49 ± 0.78 vs. 5.93 ± 0.90; 6.49 ± 0.78 vs. 6.00 ± 0.96, Δ L/min) (p = 0.006, p = 0.014), the stroke volume (stroke volume)decreased significantly (87.90 ± 15.10 vs. 81.04 ± 16.35; 87.90 ± 15.10 vs. 79.24 ± 16.83, Δ ml/beat) (p = 0.017, p = 0.0003); heart rate increased significantly in posterior extension (75.08 ± 10.43 vs. 78.42 ± 10.18, Δ beat/min) (p = 0.001); left rotation stroke volume index (SVI) decreased significantly (47.28 ± 7.97 vs. 46.14 ± 8.06, Δ ml/m2) (p = 0.0003); in the analysis of HRV-related indicators, compared with the neutral trunk position, the LF/HF of the posterior extension was significantly increased (1.90 ± 1.38 vs. 3.00 ± 1.17, p = 0.037), and the LF/HF of the forward flexion was significantly increased (1.90 ± 1.38 vs. 2.85 ± 1.41, p = 0.041), and the frequency-domain index LF/HF of right rotation was significantly increased (1.90 ± 1.38 vs. 4.06 ± 2.19, p = 0.008). There was no significant difference in respiratory rate (p > 0.05). Conclusion: A neutral trunk is the best resting position, and deviations from a neutral trunk position can affect the cardiovascular and autonomic nervous systems, resulting in decreased stroke volume, increased heart rate, and relative activation of sympathetic tone.

Singapore Grouper Iridovirus VP131 Drives Degradation of STING-TBK1 Pathway Proteins and Negatively Regulates Antiviral Innate Immunity.

Iridoviruses are large DNA viruses which cause great economic losses to the aquaculture industry and serious threats to ecological diversity worldwide. Singapore grouper iridovirus (SGIV), a novel member of the genus Ranavirus, causes high mortality in grouper aquaculture. Previous work on genome annotation demonstrated that SGIV contained numerous uncharacterized or hypothetical open reading frames (ORFs), whose functions remained largely unknown. Here, we reported that the protein encoded by SGIV ORF131R (VP131) was localized predominantly within the endoplasmic reticulum (ER). Ectopic expression of GFP-VP131 significantly enhanced SGIV replication, while VP131 knockdown decreased viral infection in vitro, suggesting that VP131 functioned as a proviral factor during SGIV infection. Overexpression of GFP-VP131 inhibited the interferon (IFN)-1 promoter activity and mRNA level of IFN-related genes induced by poly(I:C), Epinephelus coioides cyclic GMP/AMP synthase (EccGAS)/stimulator of IFN genes (EcSTING), TANK-binding kinase 1 (EcTBK1), or melanoma differentiation-associated gene 5 (EcMDA5), whereas such activation induced by mitochondrial antiviral signaling protein (EcMAVS) was not affected. Moreover, VP131 interacted with EcSTING and degraded EcSTING through both the autophagy-lysosome pathway and ubiquitin-proteasome pathway, and targeted for the K63-linked ubiquitination. Of note, we also found that EcSTING significantly accelerated the formation of GFP-VP131 aggregates in co-transfected cells. Finally, GFP-VP131 inhibited EcSTING- or EcTBK1-induced antiviral activity upon red-spotted grouper nervous necrosis virus (RGNNV) infection. Together, our results demonstrated that the SGIV VP131 negatively regulated the IFN response by inhibiting EcSTING-EcTBK1 signaling for viral evasion. IMPORTANCE STING has been identified as a critical factor participating in the innate immune response which recruits and phosphorylates TBK1 and IFN regulatory factor 3 (IRF3) to induce IFN production and defend against viral infection. However, viruses also distort the STING-TBK1 pathway to negatively regulate the IFN response and facilitate viral replication. Here, we reported that SGIV VP131 interacted with EcSTING within the ER and degraded EcSTING, leading to the suppression of IFN production and the promotion of SGIV infection. These results for the first time demonstrated that fish iridovirus evaded the host antiviral response via abrogating the STING-TBK1 signaling pathway.

Association between Handgrip Strength and the Systemic Immune-Inflammation Index: A Nationwide Study, NHANES 2011-2014.

(1) Background: The Systemic immune-inflammatory index (SII) has been proven to be an effective biomarker of human immune and inflammatory levels and has prognostic significance for most diseases. Handgrip strength (HGS) is a simple and low-cost strength measurement method, which is not only highly correlated with overall muscle strength but also accurately and reliably predicts the risk of multiple chronic diseases and mortality; (2) Purpose: Association between HGS and the SII is unclear. The purpose of this study was to investigate the association between HGS and the SII in American adults; (3) Methods: We used the data from the 2011-2012 and 2013-2014 cycles of the National Health and Nutrition Examination Survey (NHANES), involving a total of 8232 American adults (aged 18-80 years). The SII was calculated as the Platelet count × Neutrophil count/Lymphocyte count; HGS was recorded as the ratio of the sum of the highest grip-strength values of each hand to body mass index taken as the relative grip strength. A weighted generalized linear regression model and analysis of restricted cubic spline regression, adjusted for confounding factors, were used in this study to assess associations between HGS and the SII in American adults; (4) Results: There was a negative correlation between the HGS and the SII of different sexes (p < 0.05), and there was a significant negative nonlinear relationship between the HGS and the SII in males (p for nonlinear = 0.0035), and the SII showed a downward trend with the increase in the HGS in males (Q2: ß = -61.03, p = 0.01; Q3: ß = -61.28, p = 0.04, Q4: ß = -64.36, p = 0.03, p for trend = 0.04), when the HGS exceeds 3.16, with the HGS increasing, the downward trend of increasing the SII slowed down. The nonlinear relationship between the HGS and the SII in females was not significant (p for nonlinear = 0.1011), and the SII showed a linear downward trend with the increase in the HGS (Q2: ß = -24.91, p = 0.25; Q3: ß = -62.01, p = 0.03, Q4: ß = -74.94, p = 0.03, p for trend = 0.01); (5) Conclusions: HGS is inversely and independently associated with SII levels, and although the limited cubic spline regression analysis showed gender differences, the overall trend of the HGS and the SII in different genders was consistent, with both showing that the SII decreased with increasing the HGS. In addition, HGS has high general applicability based on its ease of measurement; it is possible to understand one's own grip-strength level through routine grip-strength tests, and to make preliminary predictions on the current level of immunity and inflammation in the body.

Enhancing instantaneous oxygen uptake estimation by non-linear model using cardio-pulmonary physiological and motion signals.

Oxygen uptake (VO2) is an important parameter in sports medicine, health assessment and clinical treatment. At present, more and more wearable devices are used in daily life, clinical treatment and health care. The parameters obtained by wearables have great research potential and application prospect. In this paper, an instantaneous VO2 estimation model based on XGBoost was proposed and verified by using data obtained from a medical-grade wearable device (Beijing SensEcho) at different posture and activity levels. Furthermore, physiological characteristics extracted from single-lead electrocardiogram, thoracic and abdominal respiration signal and tri-axial acceleration signal were studied to optimize the model. There were 29 healthy volunteers recruited for the study to collect data while stationary (lying, sitting, standing), walking, Bruce treadmill test and recuperating with SensEcho and the gas analyzer (Metalyzer 3B). The results show that the VO2 values estimated by the proposed model are in good agreement with the true values measured by the gas analyzer (R2 = 0.94 ± 0.03, n = 72,235), and the mean absolute error (MAE) is 1.83 ± 0.59 ml/kg/min. Compared with the estimation method using a separate heart rate as input, our method reduced MAE by 54.70%. At the same time, other factors affecting the performance of the model were studied, including the influence of different input signals, gender and movement intensity, which provided more enlightenment for the estimation of VO2. The results show that the proposed model based on cardio-pulmonary physiological signals as inputs can effectively improve the accuracy of instantaneous VO2 estimation in various scenarios of activities and was robust between different motion modes and state. The VO2 estimation method proposed in this paper has the potential to be used in daily life covering the scenario of stationary, walking and maximal exercise.

Grouper interferon-induced protein 35, a CP-interacting protein, inhibits fish nodavirus replication via positively regulating host interferon and inflammatory immune response.

Interferon (IFN)-induced protein 35 (IFI35, also known as IFP35), a member of IFN induced genes (ISGs), participates in virus infection, cancer progression and the chronic inflammatory diseases. However, its roles during fish nodavirus infection still remained largely unknown. In the present study, a homolog of IFI35 from orange spotted grouper (Epinephelus coioides) (EcIFI35) was cloned and characterized. The open reading frame of EcIFI35 was composed of 1,128 bp, and encoded a 375 amino acid polypeptide, which contained two conserved N-myc-interactor (Nmi)/IFP35 domains (NIDs). Homology analysis indicated that EcIFI35 shared 95.73% and 31.96% identity with homologs of giant grouper (E. lanceolatus) and human (Homo sapiens), respectively. The transcription of EcIFI35 was significantly up-regulated in grouper spleen (GS) cells after challenged with red-spotted grouper nervous necrosis virus (RGNNV), polyinosinic:polycytidylic acid [poly(I:C)] or lipopolysaccharide (LPS). The subcellular localization analysis showed that EcIFI35 encoded a cytoplasmic protein. The ectopic expression of EcIFI35 inhibited RGNNV replication by reducing viral genes transcription and protein synthesis. Co-immunoprecipitation (Co-IP) assay demonstrated that EcIFI35 interacted with RGNNV coat protein (CP), and partly co-localized with CP. EcIFI35 overexpression promoted the expression of IFN-related molecules and pro-inflammatory factors, including IFN regulatory factor 7 (IRF7), mitochondrial antiviral signaling protein (MAVS) and myxovirus resistance gene I (MxI), nuclear factor κB (NF-κB), interleukin 6 (IL-6) and IL-8. Together, our results revealed that EcIFI35 interacted with CP and inhibited fish nodavirus replication through positively regulated host innate immune response.

Ultrasensitive electrochemiluminescence aptasensor based on ABEI reduced silver nanoparticles for the detection of profenofos.

An ultrasensitive electrochemiluminescence (ECL) aptasensor for detection of profenofos was constructed by the reducibility and chemiluminescence property of N-(aminobutyl)-N-(ethylisoluminol) (ABEI). ABEI was used to reduce silver nitrate (AgNO3) to silver nanoparticles (AgNPs), which could be adsorbed on the lattice of graphene oxide (GO) to form ABEI-AgNPs-GO complex. This compound could achieve excellent luminescence. The aptamer (Apt) modified (5') by sulfhydryl groups could be immobilized on AgNPs to capture profenofos. When profenofos was present, the ECL signal of the aptasensor would be weakened. To further demonstrate the successful construction of the aptasensor, cyclic voltammetry tests were performed on an electrochemical workstation and an ECL analyzer, respectively. The standard curve and specificity experiment both showed that the sensor had the advantages of low limit of detection (LOD) and good specificity. Under the optimal conditions, the aptasensor had a good linear response for profenofos in the range of 1 × 10-1-1 × 104 ng/mL. It also had a LOD of 6.7 × 10-2 ng/mL and a correlation coefficient (R2) of 0.9991. The aptasensor had been successfully applied to the detection of profenofos in vegetables. The recovery range of the proposed ECL aptasensor was 98 % ~ 107.4 %.

[Analysis of the Characteristics of the Outcomes of Job-Transfer Subspecialty Training Program in Pediatrics in Sichuan].

Objective: To discuss the characteristics of physician trainee outcomes after completion of the job-transfer subspecialty training in pediatrics, a program designed to increase the number of pediatricians, in Sichuan Province and to provide countermeasures for alleviating the shortage of pediatricians. Methods: We collected with questionnaire surveys information on changes in the workload and salaries experienced by physicians who completed the job-transfer subspecialty training program in pediatrics between February 2017 and May 2020 in Sichuan Province. Then, we compared the characteristics of physicians who successful became pediatricians and those who did no. Results: A total of 208 physicians completed the job-transfer subspecialty training program in pediatrics. Among them, 178, accounting for 85.6%, completed the questionnaire survey, and 120, accounting for 67.4%, had a background in other subspecialties than pediatrics. The majority (>90%) of physicians who participated in the training program came from secondary or lower levels of hospitals from the cities and prefectures all over Sichuan Province. In this study, we found that the rate of successful job transfer from being a physician to being a pediatrician in Sichuan Province in the past four years was 85.0% (102/120), with the year-by-year results being 88.2% (15/17) in 2017, 72.7% (16/22) in 2018, 86.7% (39/45) in 2019, and 94.% (32/34) in 2020. There was no significant difference between physicians who had successful job transfer and became pediatricians and those who failed to do so in terms of gender, age, hospital level, specialization prior to the job transfer, whether or not the hospital had a pediatrics department, amount of support for the pediatrics department, whether or not the physician was working at a new hospital after the job transfer, salaries, and changes of responsibilities during COVID-19 (all P>0.05). There was significant difference in the change of workload after completion of the training program between physicians who had successful job transfer and became pediatricians and those who failed to do so ( χ 2=9.037, P=0.003), and 78.4% of the trainees stated that their workload had increased after the job transfer. There was a moderate correlation between successful job transfer and changes in workload after the transfer (|Phi[ψ] |=0.729). Conclusions: The policy of government-supported job-transfer subspecialty training in pediatrics has played an active and important role in the swift resolution of the shortage of pediatricians. However, finding the root cause of and addressing the problem of the overwhelming workload of pediatricians remain challenging issues to be resolved.

Construction of a dual-model aptasensor based on G-quadruplexes generated via rolling circle amplification for visual/sensitive detection of kanamycin.

A dual-model colorimetric and electrochemical aptasensor was designed using a large number of G-quadruplexes generated by rolling circle amplification (RCA). Specific binding between target and aptamer during RCA yielded large numbers of G-quadruplexes. A colorimetric sensor was fabricated based on the interaction between the G-quadruplex and hemin, which altered the 3,3',5,5'-Tetramethylbenzidine (TMB)-catalyzed color reaction and facilitated the visual and semi-quantitative detection of kanamycin. An electrochemical sensor was constructed based on the strong interaction between the G-quadruplex and the methylene blue electrical signal molecule. Combining nanocomposites multi-walled carbon nanotubes-chitosan/gold nanoparticles (MWCNTs-CS/AuNPs) and RCA realized double-amplified electrochemical signals. Under optimized conditions, a linear relationship was obtained as the logarithm of different concentrations of kanamycin (KAN). The colorimetric aptasensor had a linear range of 1 × 102 nM to 1 × 103 nM with a detection limit of 1.949 nM. The electrochemical aptasensor had wider a linear range from 1 × 10-3 nM to 2.5 × 103 nM and a lower detection limit of 0.333 pM. The sensor combined the advantages of simple colorimetric visualization with the ultra-precision of electrochemical methods. Aptasensor showed good specificity and prevented interference. Furthermore, the prepared dual-model aptasensor facilitated the practical monitoring of KAN in milk.

The effect of the problem based learning teaching model combined with mind mapping on nursing teaching: A meta-analysis.

BACKGROUND: To evaluate the effect of the problem-based learning (PBL) teaching model combined with mind mapping on nursing teaching. METHODS: The Cochrane Library, PubMed/MEDLINE, EBSCO, OVID, Science Citation Index (SCI), China Knowledge Resource Integrated (CNKI), VIP, WF and other databases were searched for studies published from 2000 to 2021 on the application of the PBL model in nursing teaching. Literature screening and data extraction were conducted by two independent evaluators based on inclusion and exclusion criteria, and heterogeneity was analysed. A meta-analysis was performed using RevMan 5.3 software. RESULTS: A total of 1907 papers were initially retrieved, and 8 randomized controlled studies were eventually included in the meta-analysis after screening. Theoretical test scores, practical test scores, and students' self-learning ability in the experimental group were all significantly higher than those in the control group (P < 0.05); however, practical test scores were highly heterogeneous (I2 = 76%). CONCLUSION: The PBL teaching model combined with mind mapping works well for nursing teaching in China. It can improve students' self-learning ability, which is conducive to Chinese nursing students mastering theoretical knowledge and improving practical abilities, and can be affected by a variety of factors in practical teaching.

Grouper USP12 exerts antiviral activity against nodavirus infection.

The ubiquitin-specific proteases (USPs) have attracted particular attention due to their multiple functions in different biological processes. USP12, a member of the USP family, has been demonstrated to exert critical roles in diverse cellular processes, including cell death, cancer and antiviral immunity. Here, we cloned a USP12 homolog from orange spotted grouper (Epinephelus coioides, E. coioides), and its roles in fish RNA virus replication were investigated. EcUSP12 contained a 1119-bp open reading frame (ORF) encoding a 372-amino acid polypeptide, which shared 100.00% and 91.32% identity with USP12 homolog of Etheostoma cragini and Homo sapiens, respectively. Sequence analysis indicated that EcUSP12 contained a conserved peptidase-C19G domain (aa 40-369). qPCR analysis showed that EcUSP12 transcript was most abundant in head kidney and spleen of grouper E. coioides. The expression of EcUSP12 was significantly upregulated in grouper spleen (GS) cells in response to red-spotted grouper nervous necrosis virus (RGNNV) infection. Subcellular localization analysis showed that EcUSP12 was evenly distributed throughout the cytoplasm, and mainly co-localized with endoplasmic reticulum (ER). Interestingly, during RGNNV infection, the endogenous distribution of EcUSP12 was obviously altered, and mostly overlapped with viral coat protein (CP). Co-Immunoprecipitation (Co-IP) assay indicated that EcUSP12 interacted with viral CP. In addition, overexpression of EcUSP12 significantly inhibited the replication of RGNNV in vitro, as evidenced by the decrease in viral gene transcription and protein synthesis during infection. Consistently, knockdown of EcUSP12 by small interfering RNA (siRNA) promoted the replication of RGNNV. Furthermore, EcUSP12 overexpression also increased the transcription level of inflammatory factors and interferon-related genes, including tumor necrosis factor α (TNF-α), interleukin (IL)-1ß, IL-6, IL-8, interferon regulatory factor 3 (IRF3), and IRF7. Taken together, our results demonstrated that EcUSP12, as a positive regulator of IFN signaling, interacted with viral CP to inhibit virus infection.

Microglial replacement in the aged brain restricts neuroinflammation following intracerebral hemorrhage.

Aged microglia display augmented inflammatory activity after neural injury. Although aging is a risk factor for poor outcome after brain insults, the precise impact of aging-related alterations in microglia on neural injury remains poorly understood. Microglia can be eliminated via pharmacological inhibition of the colony-stimulating factor 1 receptor (CSF1R). Upon withdrawal of CSF1R inhibitors, microglia rapidly repopulate the entire brain, leading to replacement of the microglial compartment. In this study, we investigated the impact of microglial replacement in the aged brain on neural injury using a mouse model of intracerebral hemorrhage (ICH) induced by collagenase injection. We found that replacement of microglia in the aged brain reduced neurological deficits and brain edema after ICH. Microglial replacement-induced attenuation of ICH injury was accompanied with alleviated blood-brain barrier disruption and leukocyte infiltration. Notably, newly repopulated microglia had reduced expression of IL-1ß, TNF-α and CD86, and upregulation of CD206 in response to ICH. Our findings suggest that replacement of microglia in the aged brain restricts neuroinflammation and brain injury following ICH.

Burnout, depression, anxiety and insomnia among medical staff during the COVID-19 epidemic in Shanghai.

Background: Coronavirus disease 2019 (COVID-19) has progressively impacted our daily lives, resulting in unexpected physical and mental stress on medical staff. This study is designed to investigate the levels of and risk factors for burnout, depression, anxiety, and insomnia among medical staff during the COVID-19 epidemic breakout in Shanghai, China. Methods: This cross-sectional survey was conducted from May 1 to May 31, 2022, among medical staff who were on the frontline during the epidemic breakout in Shanghai from different institutions. The MBI-HSS was used to assess burnout, PHQ-9, GAD-7 and ISI were used to evaluate mental status and insomnia. Results: A total of 543 valid questionnaires were collected. The depersonalization, depression, anxiety, and insomnia scores of medical staff were significantly higher during the pandemic in Shanghai compared with norms, while lack of personal achievement scores were decreased. Working time, work unit, work environment and age are important influencers of burnout, depression and anxiety of medical staff. Long working hours are the most likely causes of burnout and emotional disorders. Medical staff in primary hospitals were most likely to suffer from burnout and emotional disorders, while medical staff in tertiary hospitals had a reduced sense of personal achievement. Young medical staff are prone to negative emotions such as depression and anxiety, while older medical staff have a lower sense of personal accomplishment. Medical staff who were not in the shelter hospitals or designated hospitals were more likely to have problems of emotional exhaustion, depersonalization and anxiety than those who were in the shelter hospitals or designated hospitals. Contracting COVID-19 had no effect on medical staff. Emotional exhaustion and depersonalization were positively correlated with anxiety, depression, and sleep disorders while personal achievement was negatively correlated with these factors. Conclusion: Medical staff in Shanghai had high burnout, depression, anxiety and insomnia levels during the epidemic outbreak in Shanghai. During the COVID-19, medical staff may suffer different psychological problems which should be concerned. Care and supports about burnout, mental health and insomnia need to be taken to promote the mental health of medical staff according to different characteristics of medical staff.