bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rapid screening of high expressing Escherichia coli colonies using a novel dicistronic-autoinducible system
Authors: Shariati Fatemeh Sadat , Norouzian Dariush , Valizadeh Vahideh , Ahangari Cohan Reza , Keramati Malihe 
Published: December 2021

Abstract
Background
Identification of high-expressing colonies is one of the main concerns in the upstream process of recombinant protein development. The common method to screen high-producing colonies is SDS-PAGE, a laborious and time-consuming process, which is based on a random and qualitative way. The current study describes the design and development of a rapid screening system composed of a dicistronic expression system containing a reporter (enhanced green fluorescent protein, eGFP), protein model (staphylokinase, SAK), and a self-inducible system containing heat shock protein 27 (Hsp27).

Results
Dicistronic-autoinducible system expressed eGFP and SAK successfully in 5-ml and 1-L culture volumes. High expressing colonies were identified during 6 h via fluorescent signals. In addition, the biological activity of the protein model was confirmed semi-quantitatively and quantitatively through radial caseinolytic and chromogenic methods, respectively. There was a direct correlation between eGFP fluorescent intensity and SAK activity. The correlation and linearity of expression between the two genes were respectively confirmed with Pearson correlation and linear regression. Additionally, the precision, limit of detection (LOD), and limit of quantification (LOQ) were determined. The expression of eGFP and SAK was stable during four freeze–thaw cycles. In addition, the developed protocol showed that the transformants can be inoculated directly to the culture, saving time and reducing the error-prone step of colony picking.

Conclusion
The developed system is applicable for rapid screening of high-expressing colonies in most research laboratories. This system can be investigated for other recombinant proteins expressed in E. coli with a potential capability for automation and use at larger scales.
Activating transcription factor 5 (ATF5) promotes tumorigenic capability and activates the Wnt/b-catenin pathway in bladder cancer
Authors: Zhou Junhao , Tian Hu , Zhi Xi , Xiao Zhuoyu , Chen Taoyi , Yuan Haoyu , Chen Qi , Chen Mingkun , Yang Jiankun , Zhou Qizhao , Xue Kangyi , Guo Wenbin , Xia Ming , Bao Jiming , Yang Cheng , Duan Haifeng , Wang Hongyi , Huang Zhipeng , Zhu Ting , Liu Cundong 
Published: December 2021

Abstract
Background
In bladder cancer, up to 70% of patients will relapse after resection within 5 years, in which the mechanism underlying the recurrence remains largely unclear.

Methods
Quantitative real-time PCR, western blot and immunohistochemistry were conducted. The assays of tumor sphere formation and tumor xenograft were further performed to assess the potential biological roles of ATF5 (activating transcription factor 5). Chromatin immunoprecipitation-qPCR and luciferase activity assays were carried out to explore the potential molecular mechanism. A two-tailed paired Student's t-test, χ2 test, Kaplan Meier and Cox regression analyses, and Spearman's rank correlation coefficients were used for statistical analyses.

Results
ATF5 is elevated in bladder urothelial cancer (BLCA) tissues, especially in recurrent BLCA, which confers a poor prognosis. Overexpressing ATF5 significantly enhanced, whereas silencing ATF5 inhibited, the capability of tumor sphere formation in bladder cancer cells. Mechanically, ATF5 could directly bind to and stimulate the promoter of DVL1 gene, resulting in activation of Wnt/β-catenin pathway.

Conclusions
This study provides a novel insight into a portion of the mechanism underlying high recurrence potential of BLCA, presenting ATF5 as a prognostic factor or potential therapeutic target for preventing recurrence in BLCA.
Public transcriptome database-based selection and validation of reliable reference genes for breast cancer research
Authors: Song Qiang , Dou Lu , Zhang Wenjin , Peng Yang , Huang Man , Wang Mengyuan 
Published: December 2021

Abstract
Background
Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is the most sensitive technique for evaluating gene expression levels. Choosing appropriate reference genes (RGs) is critical for normalizing and evaluating changes in the expression of target genes. However, uniform and reliable RGs for breast cancer research have not been identified, limiting the value of target gene expression studies. Here, we aimed to identify reliable and accurate RGs for breast cancer tissues and cell lines using the RNA-seq dataset.

Methods
First, we compiled the transcriptome profiling data from the TCGA database involving 1217 samples to identify novel RGs. Next, ten genes with relatively stable expression levels were chosen as novel candidate RGs, together with six conventional RGs. To determine and validate the optimal RGs we performed qRT-PCR experiments on 87 samples from 11 types of surgically excised breast tumor specimens (n = 66) and seven breast cancer cell lines (n = 21). Five publicly available algorithms (geNorm, NormFinder, ΔCt method, BestKeeper, and ComprFinder) were used to assess the expression stability of each RG across all breast cancer tissues and cell lines.

Results
Our results show that RG combinations SF1 + TRA2B + THRAP3 and THRAP3 + RHOA + QRICH1 showed stable expression in breast cancer tissues and cell lines, respectively, and that they displayed good interchangeability. We propose that these combinations are optimal triplet RGs for breast cancer research.

Conclusions
In summary, we identified novel and reliable RG combinations for breast cancer research based on a public RNA-seq dataset. Our results lay a solid foundation for the accurate normalization of qRT-PCR results across different breast cancer tissues and cells.
A thermoreversible antibacterial zeolite-based nanoparticles loaded hydrogel promotes diabetic wound healing via detrimental factor neutralization and ROS scavenging
Authors: Qi Yao , Qian Kun , Chen Jin , E Yifeng , Shi Yijie , Li Hongdan , Zhao Liang 
Published: December 2021

Abstract
Background
As recovery time of diabetic wound injury is prolonged by the production of detrimental factors, including reactive oxygen species (ROS) and inflammatory cytokines, attenuating the oxidative stress and inflammatory reactions in the microenvironment of the diabetic wound site would be significant.

Experimental design
In our study, we prepared thermoreversible, antibacterial zeolite-based nanoparticles loaded hydrogel to promote diabetic wound healing via the neutralization of detrimental factors such as inflammatory cytokines and ROS.

Results
The cerium (Ce)-doped biotype Linde type A (LTA) zeolite nanoparticles synergistically eliminated mitochondrial ROS and neutralized free inflammatory factors, thus remodeling the anti-inflammatory microenvironment of the wound and enhancing angiogenesis. Moreover, the thermoreversible hydrogel composed of Pluronic F127 and chitosan demonstrated strong haemostatic and bactericidal behavior.

Conclusions
In conclusion, the obtained thermoreversible, antibacterial, zeolite-based nanoparticles loaded hydrogels represent a multi-targeted combination therapy for diabetic wound healing.

Graphical Abstract
A radioenhancing nanoparticle mediated immunoradiation improves survival and generates long-term antitumor immune memory in an anti-PD1-resistant murine lung cancer model
Authors: Hu Yun , Paris Sébastien , Barsoumian Hampartsoum , Abana Chike O. , He Kewen , Sezen Duygu , Wasley Mark , Masrorpour Fatemeh , Chen Dawei , Yang Liangpeng , Dunn Joe D. , Gandhi Saumil , Nguyen Quynh-Nhu , Cortez Maria Angelica , Welsh James W. 
Published: December 2021

Abstract
Background
Combining radiotherapy with PD1 blockade has had impressive antitumor effects in preclinical models of metastatic lung cancer, although anti-PD1 resistance remains problematic. Here, we report results from a triple-combination therapy in which NBTXR3, a clinically approved nanoparticle radioenhancer, is combined with high-dose radiation (HDXRT) to a primary tumor plus low-dose radiation (LDXRT) to a secondary tumor along with checkpoint blockade in a mouse model of anti-PD1-resistant metastatic lung cancer.

Methods
Mice were inoculated with 344SQR cells in the right legs on day 0 (primary tumor) and the left legs on day 3 (secondary tumor). Immune checkpoint inhibitors (ICIs), including anti-PD1 (200 μg) and anti-CTLA4 (100 μg) were given intraperitoneally. Primary tumors were injected with NBTXR3 on day 6 and irradiated with 12-Gy (HDXRT) on days 7, 8, and 9; secondary tumors were irradiated with 1-Gy (LDXRT) on days 12 and 13. The survivor mice at day 178 were rechallenged with 344SQR cells and tumor growth monitored thereafter.

Results
NBTXR3  +  HDXRT  +  LDXRT  +  ICIs had significant antitumor effects against both primary and secondary tumors, improving the survival rate from 0 to 50%. Immune profiling of the secondary tumors revealed that NBTXR3  +  HDXRT  +  LDXRT increased CD8 T-cell infiltration and decreased the number of regulatory T (Treg) cells. Finally, none of the re-challenged mice developed tumors, and they had higher percentages of CD4 memory T cells and CD4 and CD8 T cells in both blood and spleen relative to untreated mice.

Conclusions
NBTXR3 nanoparticle in combination with radioimmunotherapy significantly improves anti-PD1 resistant lung tumor control via promoting antitumor immune response.

Graphical Abstract
Development and validation of a guideline on sexual and reproductive health services for polycystic ovary syndrome in Iran: a mixed-methods study protocol
Authors: Kalhor Mehri , Mohammadi Eesa , Shahali Shadab , Amini Leila , Moghaddam-Banaem Lida 
Published: December 2021

AbstractBackgroundSexual and reproductive health (SRH) is an important aspect of women's health. Polycystic ovary syndrome is a common disease among women and has long-term negative effects on women’s health. Evidence shows that polycystic ovary syndrome has different impacts on SRH needs among women. The aim of this study is to design and validate an SRH services guideline for healthcare providers in treating women with polycystic ovary syndrome.MethodsThe guideline will be developed and validated using an exploratory sequential mixed-methods approach in three phases based on the National Institute for Health and Care Excellence (NICE) model: (1) scoping phase (describing the SRH needs of women with polycystic ovary syndrome from the results of both review and qualitative studies); (2) development phase (developing a primary guideline for SRH services); (3) validation phase (validation of the guideline will be performed by a panel of experts and stakeholders using the AGREE [Appraisal of Guidelines for Research and Evaluation] tool).DiscussionA specific and practical guideline on the SRH of Iranian women with polycystic ovary syndrome will be developed, which will be compatible with their specific needs and culture, considering the limited resources available. It will help service providers identify and address the specific needs of women with polycystic ovary syndrome.
Recommendations to improve insurance coverage for physiotherapy services in Iran: a multi criteria decision-making approach
Authors: Shahabi Saeed , Skempes Dimitrios , Behzadifar Masoud , Tabrizi Reza , Nazari Behrooz , Ghanbari Mahboubeh Khaton , Heydari Seyed Taghi , Zarei Leila , Lankarani Kamran Bagheri 
Published: December 2021

Abstract
Background
High toll of traffic-related injuries, climate change, natural disasters, population aging, as well as chronic diseases have all made considerable demands on receiving physiotherapy services in Iran. Nevertheless, there is an assortment of complications facing utilization of such services, particularly poor insurance coverage. Therefore, the present study investigated and identified gaps in insurance coverage in order to inform future policy reforms and the design of a more comprehensive and universal benefits package for physiotherapy services in Iran.

Methods
This project was carried out in Iran, using a mix-methods (viz. qualitative-quantitative) approach. Within the first phase, a qualitative study was completed to find policy recommendations. Such recommendations were then prioritized through the Analytical Hierarchy Process (AHP), in the second phase, based on effectiveness, acceptability, cost, fairness, feasibility, and time.

Results
Within the first phase, a total number of 30 semi-structured interviews with health policy-makers, health insurers, faculty members, rehabilitation experts, and physiotherapists were completed. Several policy recommendations were also proposed by the study participants. Following the second phase, prioritized recommendations were provided to promote stewardship (e.g., informing policy-makers about physiotherapy services), collection of funds (e.g., placing value-added taxes on luxury goods and services), pooling of funds (e.g., moving allocated resources towards insurance (viz. third-party) mechanism), purchasing (e.g., using strategic purchasing), and benefit package (e.g., considering preventive interventions) as the main components of insurance coverage.

Conclusion
The study findings provided a favorable ground to improve insurance coverage for physiotherapy services in Iran. As well, decision- and policy-makers can place these recommendations on the agenda in the health sector to protect population health status, especially that of groups with disabilities.
A systematic review of artificial intelligence chatbots for promoting physical activity, healthy diet, and weight loss
Authors: Oh Yoo Jung , Zhang Jingwen , Fang Min-Lin , Fukuoka Yoshimi 
Published: December 2021

Abstract
Background
This systematic review aimed to evaluate AI chatbot characteristics, functions, and core conversational capacities and investigate whether AI chatbot interventions were effective in changing physical activity, healthy eating, weight management behaviors, and other related health outcomes.

Methods
In collaboration with a medical librarian, six electronic bibliographic databases (PubMed, EMBASE, ACM Digital Library, Web of Science, PsycINFO, and IEEE) were searched to identify relevant studies. Only randomized controlled trials or quasi-experimental studies were included. Studies were screened by two independent reviewers, and any discrepancy was resolved by a third reviewer. The National Institutes of Health quality assessment tools were used to assess risk of bias in individual studies. We applied the AI Chatbot Behavior Change Model to characterize components of chatbot interventions, including chatbot characteristics, persuasive and relational capacity, and evaluation of outcomes.

Results
The database search retrieved 1692 citations, and 9 studies met the inclusion criteria. Of the 9 studies, 4 were randomized controlled trials and 5 were quasi-experimental studies. Five out of the seven studies suggest chatbot interventions are promising strategies in increasing physical activity. In contrast, the number of studies focusing on changing diet and weight status was limited. Outcome assessments, however, were reported inconsistently across the studies. Eighty-nine and thirty-three percent of the studies specified a name and gender (i.e., woman) of the chatbot, respectively. Over half (56%) of the studies used a constrained chatbot (i.e., rule-based), while the remaining studies used unconstrained chatbots that resemble human-to-human communication.

Conclusion
Chatbots may improve physical activity, but we were not able to make definitive conclusions regarding the efficacy of chatbot interventions on physical activity, diet, and weight management/loss. Application of AI chatbots is an emerging field of research in lifestyle modification programs and is expected to grow exponentially. Thus, standardization of designing and reporting chatbot interventions is warranted in the near future.

Systematic review registration
International Prospective Register of Systematic Reviews (PROSPERO): CRD42020216761.
Universal coverage of the first antenatal care visit but poor continuity of care across the maternal and newborn health continuum among Nepalese women: analysis of levels and correlates
Authors: Khatri Resham B , Karkee Rajendra , Durham Jo , Assefa Yibeltal 
Published: December 2021

Abstract
Background
Routine maternity care visits (MCVs) such as antenatal care (ANC), institutional delivery, and postnatal care (PNC) visits are crucial to utilisation of maternal and newborn health (MNH) interventions during pregnancy-postnatal period. In Nepal, however, not all women complete these routine MCVs. Therefore, this study examined the levels and correlates of (dis)continuity of MCVs across the antenatal-postnatal period.

Methods
We conducted further analysis of the 2016 Nepal Demographic and Health Survey. A total of 1,978 women aged 15–49 years, who had live birth two years preceding the survey, were included in the analysis. The outcome variable was (dis)continuity of routine MCVs (at least four ANC visits, institutional delivery, and PNC visit) across the pathway of antennal through to postnatal period. Independent variables included several social determinants of health under structural, intermediary, and health system domains. Multinomial logistic regression was conducted to identify the correlates of routine MCVs. Relative risk ratios (RR) were reported with 95% confidence intervals at a significance level of p<0.05.

Results
Approximately 41% of women completed all routine MCVs with a high proportion of discontinuation around childbirth. Women of disadvantaged ethnicities, from low wealth status, who were illiterate had higher RR of discontinuation of MCVs (compared to completion of all MCVs). Similarly, women who speak Bhojpuri, from remote provinces (Karnali and Sudurpaschim), who had a high birth order (≥4), who were involved in the agricultural sector, and who had unwanted last birth had a higher RR of discontinuation of MCVs. Women discontinued routine MCVs if they had poor awareness of health mother-groups and perceived the problem of not having female providers.

Conclusions
Routine monitoring using composite coverage indicators is required to track the levels of (dis)continuity of routine MCVs at the maternity care continuum. Strategies such as raising awareness on the importance of maternity care, care provision from female health workers could potentially improve the completion of MCVs. In addition, policy and programmes for continuity of maternity care are needed to focus on women with socioeconomic and ethnic disadvantages and from remote provinces.
Mechanism of drug-induced liver injury and hepatoprotective effects of natural drugs
Authors: Zhou Yongfeng , Wang Junnan , Zhang Dingkun , Liu Jiaxin , Wu Qinghua , Chen Jiang , Tan Peng , Xing Boyu , Han Yanzhong , Zhang Ping , Xiao Xiaohe , Pei Jin 
Published: December 2021

AbstractDrug-induced liver injury (DILI) is a common adverse drug reaction (ADR) and a serious threat to health that affects disease treatments. At present, no targeted clinical drugs are available for DILI. Traditional natural medicines have been widely used as health products. Some natural medicines exert specific hepatoprotective effects, with few side effects and significant clinical efficacy. Thus, natural medicines may be a promising direction for DILI treatment. In this review, we summarize the current knowledge, common drugs and mechanisms of DILI, as well as the clinical trials of natural drugs and their bioactive components in anticipation of the future development of potential hepatoprotective drugs.
Testing an early online intervention for the treatment of disturbed sleep during the COVID-19 pandemic in self-reported good and poor sleepers (Sleep COVID-19): study protocol for a randomised controlled trial
Authors: Sawdon Olivia L. , Elder Greg J. , Santhi Nayantara , Alfonso-Miller Pamela , Ellis Jason G. 
Published: December 2021

Abstract
Background
Theoretical models of insomnia suggest that stressful life events, such as the COVID-19 pandemic, can cause acute insomnia (short-term disruptions to sleep). Early interventions may prevent short-term sleep problems from progressing to insomnia disorder. Although cognitive behavioural therapy for insomnia (CBT-I) is effective in treating insomnia disorder, this can be time and resource-intensive. Further, online interventions can be used to deliver treatment to a large number of individuals. The objective of this study is to investigate if an online behavioural intervention, in the form of a leaflet, which has been successfully used alongside CBT-I for acute insomnia, can reduce symptoms of acute insomnia in poor sleepers.

Methods
A total of 124 self-reported good and poor sleepers will be enrolled in an online stratified randomised controlled trial. After baseline assessments (T1), participants will complete a 1-week pre-intervention sleep monitoring period (T2) where they will complete daily sleep-diaries. Poor sleepers (n = 62) will be randomly allocated to an invention or wait-list group, where they will receive the intervention (T3), or will do so after a 28-day delay. Good sleepers (n = 62) will be randomly assigned to an intervention or no intervention group. All participants will complete a 1-week post intervention sleep monitoring period using daily sleep diaries (T4). Participants will be followed up at 1 week (T5), 1 month (T6) and 3 months (T7) post intervention. The primary outcome measure will be insomnia severity, measured using the Insomnia Severity Index. Secondary outcome measures will include subjective mood and subjective sleep continuity, measured using sleep diaries. Data will be analysed using an intention-to-treat approach.

Discussion
It is expected that this online intervention will reduce symptoms of acute insomnia in self-reported short-term poor sleepers, and will also prevent the transition to poor sleep in good sleepers. We expect that this will demonstrate the feasibility of online interventions for the treatment and prevention of acute insomnia. Specific advantages of online approaches include the low cost, ease of administration and increased availability of treatment, relative to face-to-face therapy.

Trial registration
ISRCTN43900695 (Prospectively registered 8th of April 2020).