Valorization of pawpaw (Carica papaya L.) leaves as a source of polyphenols by ionic liquid-based microwave-assisted extraction: Comparison with other extraction methods and bioactivity evaluation.

This study aimed to valorize pawpaw (Carica papaya L.) leaves as a rich source of polyphenols through the application of ionic liquid-based microwave-assisted extraction (ILMAE). Initially, the ILMAE process was optimized using response surface methodology (RSM), resulting in a total polyphenols yield of 27.84 ± 0.33 mg GAE/g DW under the optimal conditions: [BMIM]Br concentration of 0.57 mol/L, extraction time of 14 min, microwave power of 460 W, extraction temperature of 77 °C, solvent-to-material ratio of 30 mL/g, and three extraction cycles. Compared to conventional methods such as maceration extraction (ME), heat reflux extraction (HRE), and microwave-assisted extraction (MAE), the ILMAE method exhibited a significantly higher PLTP yield. Furthermore, the PLTP extracts demonstrated strong antioxidant activity against DPPH• and ABTS+• radicals, as well as a significant inhibitory effect on α-glucosidase activity. This work demonstrates that ILMAE is a green and efficient strategy for the valorization of pawpaw leaves.

Emergence of a new designated clade 16 with significant antigenic drift in hemagglutinin gene of H9N2 subtype avian influenza virus in eastern China.

H9N2 avian influenza viruses (AIVs) pose an increasing threat to the poultry industry worldwide and have pandemic potential. Vaccination has been principal prevention strategy to control H9N2 in China since 1998, but vaccine effectiveness is persistently challenged by the emergence of the genetic and/or antigenic variants. Here, we analysed the genetic and antigenic characteristics of H9N2 viruses in China, including 70 HA sequences of H9N2 isolates from poultry, 7358 from online databases during 2010-2020, and 15 from the early reference strains. Bayesian analyses based on hemagglutinin (HA) gene revealed that a new designated clade16 emerged in April 2012, and was prevalent and co-circulated with clade 15 since 2013 in China. Clade 16 viruses exhibited decreased cross-reactivity with those from clade 15. Antigenic Cartography analyses showed represent strains were classified into three antigenic groups named as Group1, Group2 and Group3, and most of the strains in Group 3 (15/17, 88.2%) were from Clade 16 while most of the strains in Group2 (26/29, 89.7%) were from Clade 15. The mean distance between Group 3 and Group 2 was 4.079 (95%CI 3.605-4.554), revealing that major switches to antigenic properties were observed over the emergence of clade 16. Genetic analysis indicated that 11 coevolving amino acid substitutions primarily at antigenic sites were associated with the antigenic differences between clade 15 and clade 16. These data highlight complexities of the genetic evolution and provide a framework for the genetic basis and antigenic characterization of emerging clade 16 of H9N2 subtype avian influenza virus.

Enhanced pathogenicity and transmissibility of H9N2 avian influenza virus in mammals by hemagglutinin mutations combined with PB2-627K.

H9N2 avian influenza viruses (AIVs) circulate globally in poultry and have become the dominant AIV subtype in China in recent years. Previously, we demonstrated that the H9N2 virus (A/chicken/Eastern China/SDKD1/2015) naturally harbors a mammalian-adaptive molecular factor (627K) in the PB2 protein and is weakly pathogenic in mice. Here, we focused on new markers for virulence in mammals. A mouse-adapted H9N2 virus was serially passaged in mice by infecting their lungs. As expected, infected mice showed clinical symptoms and died at passage six. A comparison between the wild-type and mouse-adapted virus sequences identified amino acid substitutions in the hemagglutinin (HA) protein. H9N2 viruses with the T187P â€‹+ â€‹M227L double mutation exhibited an increased affinity to human-type (SAα2,6Gal) receptors and significantly enhanced viral attachment to mouse lung tissues, which contributed to enhancing viral replication and virulence in mice. Additionally, HA with the T187P â€‹+ â€‹M227L mutation enabled H9N2 viral transmission in guinea pigs via direct contact. AIV pathogenicity in mice is a polygenic trait. Our results demonstrated that these HA mutations might be combined with PB2-627K to significantly increase H9N2 virulence in mice, and this enhanced virulence was achieved in other H9N2 AIVs by generating the same combination of mutations. In summary, our study identified novel key elements in the HA protein that are required for H9N2 pathogenicity in mice and provided valuable insights into pandemic preparedness against emerging H9N2 strains.

Dynamic Monte Carlo Simulations of Strain-Induced Crystallization in Multiblock Copolymers: Effects of Asymmetric Block Rigidity.

Thermoplastic elastomers such as polyether-b-polyamides (or -polyesters), polyurethanes (or with -urea) and olefin block copolymers are commonly processed through a stretching process for achieving high elasticity and high toughness in their products, while the size diversity of semicrystalline microdomains of hard blocks appears as the key factor. By means of dynamic Monte Carlo simulations of strain-induced crystallization of locally concentrated and diluted crystallizable blocks alternatingly connected with noncrystallizable blocks in diblock and tetrablock copolymers, we have studied the size diversity of semicrystalline microdomains presumably raised by local concentration fluctuations of crystallizable blocks and found the dilution effects to persist from diblock to tetrablock copolymers. In the present work, we continued to study the effects of asymmetric block rigidity between crystallizable and noncrystallizable blocks on strain-induced crystallization of concentrated and diluted crystallizable blocks in diblock copolymers. The results showed that when crystallizable blocks hold higher thermodynamic rigidity than noncrystallizable blocks, the large semicrystalline domains become larger and the small semicrystalline domains become more, enhancing their size diversity. However, asymmetric kinetic rigidity has little effect. Our observations imply that industrial stretching processing could enhance the toughness of semicrystalline thermoplastic elastomers when their crystallizable blocks hold a higher thermodynamic rigidity relative to noncrystallizable blocks. Our integrated approach paved the way for a better understanding of the structure-property relationship in thermoplastic elastomers.

Worker Characteristics and Measures Associated With Patient and Visitor Violence in the COVID-19 Pandemic: A Multilevel Regression Analysis From China.

Objective: To analyze the patient and visitor workplace violence (PVV) toward health workers (HWs) and identify correlations between worker characteristics, measures against violence and exposure to PVV in COVID-19 pandemic. Methods: A cross-sectional survey utilizing the international questionnaires in six public tertiary hospitals from Beijing in 2020 was conducted, and valid data from 754 respondents were collected. Multilevel logistic regression models were used to determine the association between independents and exposure to PVV. Results: During COVID-19 pandemic and regular epidemic prevention and control, doctors were 5.3 times (95% CI = 1.59~17.90) more likely to suffer from physical PVV than nurses. HWs most frequently work with infants were 7.2 times (95% CI = 2.24~23.19) more likely to suffer from psychological PVV. More than four-fifth of HWs reported that their workplace had implemented security measures in 2020, and the cross-level interactions between the security measures and profession variable indicates that doctors in the workplace without security measures were 11.3 times (95% CI = 1.09~116.39) more likely to suffer from physical PVV compared to nurses in the workplace with security measures. Conclusion: Doctors have higher risk of physical PVV in COVID-19 containment, and the security measures are very important and effective to fight against the physical PVV. Comprehensive measures should be implemented to mitigate hazards and protect the health, safety, and well-being of health workers.

The Association between Parental Marital Satisfaction and Adolescent Prosocial Behavior in China: A Moderated Serial Mediation Model.

Parental marital satisfaction is a well-established protective factor for prosocial behavior in adolescents, yet the parental socialization of adolescent prosocial behavior is limited in Chinese culture. Furthermore, it remains unknown whether the association between parental marital satisfaction and adolescent prosocial behavior is mediated by parent-child intimacy and/or adolescent empathy. The present study examined these associations in 480 students (50.4% male, age = 12.7 ± 0.54 years) and one of their parents. Path analysis (Mplus) revealed that parental marital satisfaction was related to a high level of parent-child intimacy, which was, in turn, associated with a high level of adolescent empathy, which itself predicted adolescent prosocial behavior. The serial mediation effect differed between boys and girls. Specifically, compared with boys, girls' parent-child intimacy was a stronger predictor of their empathy, and empathy had a stronger predictive effect on their prosocial behavior. Several implications for interventions and policies for increasing prosocial behavior in children are suggested.

Dynamic Monte Carlo simulations of strain-induced crystallization in multiblock copolymers: effects of dilution.

Multiblock copolymers containing alternating semicrystalline and molten blocks are good thermoplastic elastomers. Their crystallization in the stretching process is however complicated by the dilution effects, prior microphase separation and contrast chain rigidity of the molten blocks. We designed our systematic investigation with three integrated steps, and herein, as the first step, we considered only the dilution effects without prior microphase separation and contrast chain rigidity. We compared two extreme situations of local dilution separately corresponding to parallel-posited and antiparallel-posited block copolymers upon strain-induced crystallization. Our dynamic Monte Carlo simulations of diblock and tetrablock copolymers demonstrated that the stretching introduces a constraint on the diffusion of locally posited crystallizable blocks along the stretching direction for crystallization and thus enhances the dilution effects to result in a higher diversity in crystal stabilities. We observed that the strain-induced crystallization of parallel-posited copolymers behaved like the melt crystallization of homopolymers; in contrast, the strain-induced crystallization of antiparallel-posited copolymers yielded crystallites near the block junction, which are relatively small and less stable due to their local dilution suppressing their melting points. Similar to the case of spider dragline silks, two contrasting stabilities of crystallites in semicrystalline multiblock copolymers explain their good toughness. Our modeling approach paves the way toward a better understanding of the structure-property relationship in the semicrystalline thermoplastic elastomers.

Forecasting Future Demand of Nursing Staff for the Oldest-Old in China by 2025 Based on Markov Model.

BACKGROUND: An aging population and an increase in the proportion of disabled elderly have brought an unprecedented global challenge, especially in China. Aside lack of professional long-term care facilities, the shortage of human resource for old-age care is also a major threat. Therefore, this study tries to forecast the demand scale of nursing staff for the oldest-old in 2025 in China servicing as a reference for the development plan of human resource for elderly nursing. METHODS: Based on CLHLS (Chinese Longitudinal Healthy Longevity Survey) 2011 and 2014, Logit model was used to construct the transition probability matrix of the elderly's health status (health/mild/moderate/severe disability and death). By using the data of the elderly population aged 65 or over in the 2010 national population census, we projected the number of Chinese oldest-old population in different health status by 2025 through Markov model and projected the scale of the demand of nursing staff combined with the human population ratio method. RESULTS: The forecast shows that the Chinese oldest-old population is about 52.6 million, among which 46.9 million are healthy, 3.7 million are mild, 0.8 million are moderate, and 1.2 million are severely disabled in 2025. Concurrently, the demand scale of nursing staff will be 5.6 million according to the low standard and 11.5 million according to the high standard. Thus, human resource supply of long-term care is worrying. CONCLUSION: In 2025, the population size of the Chinese oldest-old will be further expanded, and the demand of care will increase accordingly, leading to a vast gap in the nursing staff. Therefore, it is urgent to build a professional nursing staff with excellent comprehensive quality and reasonable quantity, to ensure the sustainable development of China's elderly care service industry.

Effects of the joint prevention and control of atmospheric pollution policy on air pollutants-A quantitative analysis of Chinese policy texts.

Joint prevention and control of atmospheric pollution (JPCAP) policies play a vital role in alleviating regional pollution. Based on Latent Dirichlet Allocation (LDA) model, we construct two policy strength measures of effectiveness and number, and investigate the effects of policy strength on air pollutant emissions for four types of JPCAP policies. The results show that the effects of economic incentive policy tools and supporting policy tools on emission reduction deviate significantly from policy preferences. Economic incentive policy tools are the most effective in promoting emission reductions in SO2, NOx and dust, but their effectiveness are the lowest in reality. Supporting policy tools, with the highest strength, have little effect on emission reduction. Command-control policies and persuasion policies are both relatively high in quantity and effectiveness. In addition, policy strength plays a more important role in reducing air pollutants in key regions than in non-key regions. JPCAP policies have gradually changed from a single policy tool to multiple policy tools, and the government shifted its attention to improving the legal effectiveness of policies after 2015. Finally, we propose some policy implications to optimize JPCAP policies and address regional air pollution problem.

Platform study of genotyping-guided precision medicine for rare solid tumours: a study protocol for a phase II, non-randomised, 18-month, open-label, multiarm, single-centre clinical trial testing the safety and efficacy of multiple Chinese-approved targeted drugs and PD-1 inhibitors in the treatment of metastatic rare tumours.

INTRODUCTION: Limited clinical studies have been conducted on rare solid tumours, and there are few guidelines on the diagnosis and treatment, including experiences with targeted therapy and immunotherapy, of rare solid tumours in China, resulting in limited treatment options and poor outcomes. This study first proposes a definition of rare tumours and is designed to test the preliminary efficacy of targeted and immunotherapy drugs for the treatment of rare tumours. METHODS AND ANALYSIS: This is a phase II, open-label, non-randomised, multiarm, single-centre clinical trial in patients with advanced rare solid tumours who failed standard treatment; the study aims to evaluate the safety and efficacy of targeted drugs in patients with advanced rare solid tumours with corresponding actionable alterations, as well as the safety and efficacy of immune checkpoint (programmed death receptor inhibitor 1, PD-1) inhibitors in patients with advanced rare solid tumours without actionable alterations. Patients with advanced rare tumours who fail standardised treatment and carry actionable alterations (Epidermal growth factor receptor (EGFR) mutations, ALK gene fusions, ROS-1 gene fusions, C-MET gene amplifications/mutations, BRAF mutations, CDKN2A mutations, BRCA1/2 mutations, HER-2 mutations/overexpressions/amplifications or C-KIT mutations) will be enrolled in the targeted therapy arm and be given the corresponding targeted drugs. Patients without actionable alterations will be enrolled in the PD-1 inhibitor arm and be treated with sintilimab. After the patients treated with vemurafenib, niraparib and palbociclib acquire resistance, they will receive combination treatment with sintilimab or atezolizumab. With the use of Simon's two-stage Minimax design, and the sample size was estimated to be 770. The primary endpoint of this study is the objective response rate. The secondary endpoints are progression-free survival in the targeted treatment group and single-agent immunotherapy group; the duration of response in the targeted therapy and single-agent immunotherapy groups; durable clinical benefit in the single-agent immunotherapy group; and the incidence of adverse events. ETHICS AND DISSEMINATION: Ethics approval was obtained from the Chinese Academy of Medical Sciences (ID: 20/132-2328). The results from this study will be actively disseminated through manuscript publications and conference presentations. TRIAL REGISTRATION NUMBERS: NCT04423185; ChiCTR2000039310.

Peripheral Lymphoid Volume Expansion and Maintenance Are Controlled by Gut Microbiota via RALDH+ Dendritic Cells.

Lymphocyte homing to draining lymph nodes is critical for the initiation of immune responses. Secondary lymphoid organs of germ-free mice are underdeveloped. How gut commensal microbes remotely regulate cellularity and volume of secondary lymphoid organs remains unknown. We report here that, driven by commensal fungi, a wave of CD45(+)CD103(+)RALDH(+) cells migrates to the peripheral lymph nodes after birth. The arrival of these cells introduces high amounts of retinoic acid, mediates the neonatal to adult addressin switch on endothelial cells, and directs the homing of lymphocytes to both gut-associated lymphoid tissues and peripheral lymph nodes. In adult mice, a small number of these RALDH(+) cells might serve to maintain the volume of secondary lymphoid organs. Homing deficiency of these cells was associated with lymph node attrition in vitamin-A-deficient mice, suggesting a perpetual dependence on retinoic acid signaling for structural and functional maintenance of peripheral immune organs.

Regularized linear discriminant analysis and its application in microarrays.

In this paper, we introduce a modified version of linear discriminant analysis, called the "shrunken centroids regularized discriminant analysis" (SCRDA). This method generalizes the idea of the "nearest shrunken centroids" (NSC) (Tibshirani and others, 2003) into the classical discriminant analysis. The SCRDA method is specially designed for classification problems in high dimension low sample size situations, for example, microarray data. Through both simulated data and real life data, it is shown that this method performs very well in multivariate classification problems, often outperforms the PAM method (using the NSC algorithm) and can be as competitive as the support vector machines classifiers. It is also suitable for feature elimination purpose and can be used as gene selection method. The open source R package for this method (named "rda") is available on CRAN (http://www.r-project.org) for download and testing.

Global transcriptional response to interferon is a determinant of HCV treatment outcome and is modified by race.

Interferon (IFN)-alpha-based therapy for chronic hepatitis C is effective in fewer than 50% of all treated patients, with a substantially lower response rate in black patients. The goal of this study was to investigate the underlying host transcriptional response associated with interferon treatment outcomes. We collected peripheral blood mononuclear cells from chronic hepatitis C patients before initiation of IFN-alpha therapy and incubated the cells with or without IFN-alpha for 6 hours, followed by microarray assay to identify IFN-induced gene transcription. The microarray datasets were analyzed statistically according to the patients' race and virological responses to subsequent IFN-alpha treatment. The global induction of IFN-stimulated genes (ISGs) was significantly greater in sustained virological responders compared with nonresponders and in white patients compared with black patients. In addition, a significantly greater global induction of ISGs was observed in sustained virological responders compared with nonresponders within the group of white patients. The level of IFN-induced signal transducer and activator of transcription (STAT) 1 activation, a key component of the Janus kinase (JAK)-STAT signaling pathway, correlated with the global induction of ISGs and was significantly higher in white patients than in black patients. In conclusion, both treatment outcome and race are associated with different transcriptional responses to IFN-alpha. Because this difference is evident in the global induction of ISGs rather than a selective effect on a subset of such genes, key factors affecting the outcome of IFN-alpha therapy are likely to act at the JAK-STAT pathway that controls transcription of downstream ISGs.

Microarray analysis of differentially expressed genes in vaginal tissues from women with stress urinary incontinence compared with asymptomatic women.

BACKGROUND: The pathophysiology of pelvic floor dysfunction resulting in stress urinary incontinence (SUI) in women is complex. Evidence suggests that there is also a genetic predisposition towards SUI. We sought to identify differentially expressed genes involved in extracellular matrix (ECM) metabolism in vaginal tissues from women with SUI in the secretory phase of menses compared with asymptomatic women. METHODS: Tissue samples were taken from the periurethral vaginal wall of five pairs of premenopausal, age-matched SUI and continent women and subjected to microarray analysis using the GeneChip Human Genome U133 oligonucleotide chip set. RESULTS: Extensive statistical analyses generated a list of 79 differentially expressed genes. Elafin, keratin 16, collagen type XVII and plakophilin 1 were consistently identified as up-regulated ECM genes. Elafin, a serine protease inhibitor involved in the elastin degradation pathway and wound healing, was expressed in pelvic fibroblasts and confirmed by Western blot, quantitative competitive PCR and immunofluorescence cell staining. CONCLUSIONS: Genes involved in elastin metabolism were differentially expressed in vaginal tissue from women with SUI, suggesting that elastin remodelling may be important in the molecular aetiology of SUI.

Microarray analysis reveals differences in gene expression of circulating CD8(+) T cells in melanoma patients and healthy donors.

Circulating T cells from many cancer patients are known to be dysfunctional and undergo spontaneous apoptosis. We used microarray technology to determine whether gene expression differences exist in T cells from melanoma patients versus healthy subjects, which may underlie these abnormalities. To maximize the resolution of our data, we sort purified CD8(+) subsets and amplified the extracted RNA for microarray analysis. These analyses show subtle but statistically significant expression differences for 10 genes in T cells from melanoma patients versus healthy controls, which were additionally confirmed by quantitative real-time PCR analysis. Whereas none of these genes are members of the classical apoptosis pathways, several may be linked to apoptosis. To additionally investigate the significance of these 10 genes, we combined them into a classifier and found that they provide a much better discrimination between melanoma and healthy T cells as compared with a classifier built uniquely with classical apoptosis-related genes. These results suggest the possible engagement of an alternative apoptosis pathway in circulating T cells from cancer patients.