bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immortalization effect of SV40T lentiviral vectors on canine corneal epithelial cells
Authors: Guo Long , Wang Zhihao , Li Jun , Li Jianji , Cui Luying , Dong Junsheng , Meng Xia , Qian Chen , Wang Heng 
Published: May 2022

Abstract
Background
Primary canine corneal epithelial cells (CCECs) easily become senescent, and cell proliferation is limited. Therefore, sampling for experimentation requires a large number of animals, which is problematic in terms of animal welfare and fails to maintain the stability of the cells for in vitro analyses.

Results
In this study, CCECs were separated and purified by trypsin and dispase II enzymatic analysis. Next, the cells were immortalized by transfection with a lentiviral vector expressing Simian vacuolating virus 40 large T (SV40T). The immortalized canine corneal epithelial cell line (CCEC-SV40T) was established by serial passages and monoclonal selection. The biological characteristics of CCEC-SV40T cells were evaluated based on the cell proliferation rate, cell cycle pattern, serum dependence, karyotype, and cytokeratin 12 immunofluorescence detection. In addition, we infected CCEC-SV40T cells with Staphylococcus pseudintermedius (S. pseudintermedius) and detected the inflammatory response of the cells. After the CCEC-SV40T cells were passaged continuously for 40 generations, the cells grew in a cobblestone pattern, which was similar to CCECs. The SV40T gene and cytokeratin 12 can be detected in each generation. CCEC-SV40T cells were observed to have a stronger proliferation capacity than CCECs. CCEC-SV40T cells maintained the same diploid karyotype and serum-dependent ability as CCECs. After CCEC-SV40T cells were infected with S. pseudintermedius, the mRNA expression levels of NLRP3, Caspase-1 and proinflammatory cytokines, including IL-1β, IL-6, IL-8 and TNF-α, were upregulated, and the protein levels of MyD88, NLRP3 and the phosphorylation of Iκbα and p65 were upregulated.

Conclusions
In conclusion, the CCEC-SV40T line was successfully established and can be used for in vitro studies, such as research on corneal diseases or drug screening.
MiR-129-5p exerts Wnt signaling-dependent tumor-suppressive functions in hepatocellular carcinoma by directly targeting hepatoma-derived growth factor HDGF
Authors: Huge Nicole , Reinkens Thea , Buurman Reena , Sandbothe Maria , Bergmann Anke , Wallaschek Hannah , Vajen Beate , Stalke Amelie , Decker Melanie , Eilers Marlies , Schäffer Vera , Dittrich-Breiholz Oliver , Gürlevik Engin , Kühnel Florian , Schlegelberger Brigitte , Illig Thomas , Skawran Britta 
Published: May 2022

AbstractBackgroundIn hepatocellular carcinoma (HCC), histone deacetylases (HDACs) are frequently overexpressed. This results in chromatin compaction and silencing of tumor-relevant genes and microRNAs. Modulation of microRNA expression is a potential treatment option for HCC. Therefore, we aimed to characterize the epigenetically regulated miR-129-5p regarding its functional effects and target genes to understand its relevance for HCC tumorigenesis.MethodsGlobal miRNA expression of HCC cell lines (HLE, HLF, Huh7, HepG2, Hep3B) and normal liver cell lines (THLE-2, THLE-3) was analyzed after HDAC inhibition by miRNA sequencing. An in vivo xenograft mouse model and in vitro assays were used to investigate tumor-relevant functional effects following miR-129-5p transfection of HCC cells. To validate hepatoma-derived growth factor (HDGF) as a direct target gene of miR-129-5p, luciferase reporter assays were performed. Survival data andHDGFexpression were analyzed in public HCC datasets. After siRNA-mediated knockdown ofHDGF, its cancer-related functions were examined.ResultsHDAC inhibition induced the expression of miR-129-5p. Transfection of miR-129-5p increased the apoptosis of HCC cells, decreased proliferation, migration and ERK signaling in vitro and inhibited tumor growth in vivo.Direct binding of miR-129-5p to the 3′UTR ofHDGFvia a noncanonical binding site was validated by luciferase reporter assays.HDGFknockdown reduced cell viability and migration and increased apoptosis in Wnt-inactive HCC cells. These in vitro results were in line with the analysis of public HCC datasets showing thatHDGFoverexpression correlated with a worse survival prognosis, primarily in Wnt-inactive HCCs.ConclusionsThis study provides detailed insights into the regulatory network of the tumor-suppressive, epigenetically regulated miR-129-5p in HCC. Our results reveal for the first time that the therapeutic application of mir-129-5p may have significant implications for the personalized treatment of patients with Wnt-inactive, advanced HCC by directly regulatingHDGF. Therefore, miR-129-5p is a promising candidate for a microRNA replacement therapy to prevent HCC progression and tumor metastasis.
A novel nanobody as therapeutics target for EGFR-positive colorectal cancer therapy: exploring the effects of the nanobody on SW480 cells using proteomics approach
Authors: Lamtha Thomanai , Krobthong Sucheewin , Yingchutrakul Yodying , Samutrtai Pawitrabhorn , Gerner Christopher , Tabtimmai Lueacha , Choowongkomon Kiattawee 
Published: May 2022

Abstract
Background
The epidermal growth factor receptor (EGFR) overexpression is found in metastatic colorectal cancer (mCRC). Targeted molecular therapies such as monoclonal antibodies (mAbs) and tyrosine kinase inhibitors (TKI) are becoming more precise, targeting specifically for cancer therapeutics. However, there are adverse effects of currently available anti-EGFR drugs, including drug-resistant and side effects. Nanobodies can overcome these limitations. Our previous study has found that cell-penetrable nanobodies targeted at EGFR-tyrosine kinase were significantly reduced EGFR-positive lung cancer cells viability and proliferation. The aim of the present study was to determine the effect of cell-penetrable nanobody (R9VH36) on cell viability and proteomic profile in EGFR-positive human colorectal cancer cell lines.

Methods
The human colorectal carcinoma cell line (SW480) was treated with R9VH36, compared with gefitinib. Cell viability was monitored using the MTT cell viability assay. The proteomic profiling was analyzed by LC–MS/MS .

Results
The half-maximal inhibitory concentration (IC50) values determined for R9VH36 and gefitinib against SW480 were 527 ± 0.03 nM and 13.31 ± 0.02 μM, respectively. Moreover, both the gefitinib-treated group and nanobody-treated group had completely different proteome profiles. A total 6626 differentially expressed proteins were identified. PCA analysis revealed different proteome profiling in R9VH36 experiment. There were 8 proteins in R9VH36 that significantly exhibited opposite expression directions when compared to gefitinib. These proteins are involved in DNA-damage checkpoint processes.

Conclusion
The proteomics explored those 6,626 proteins had different expressions between R9VH36 and gefitinib. There were 8 proteins in R9VH36 exhibited opposite expression direction when comparing to gefitinib. Our findings suggest that R9VH36 has the potential to be an alternative remedy for treating EGFR-positive colon cancer.
Evaluating official development assistance-funded granting mechanisms for global health and development research that is initiated in high-income countries
Authors: Cassola Adèle , Baral Prativa , Røttingen John-Arne , Hoffman Steven J. 
Published: May 2022

Abstract
Background
Several countries allocate official development assistance (ODA) for research on global health and development issues that is initiated in the donor country. The integration of such research within domestic research systems aligns with efforts to coordinate ODA investments with science, technology and innovation policies towards achieving the Sustainable Development Goals (SDGs).

Methods
Through a document synthesis and interviews with research funders in ODA donor and recipient countries, we evaluated the performance of this funding approach across seven donor-country programmes from five donor countries and examined the institutional design elements that increase its chances of advancing development goals and addressing global challenges.

Results
We found that carefully designed programmes provide a promising pathway to producing valuable and contextually relevant knowledge on global health and development issues. To achieve these outcomes and ensure they benefit ODA-receiving countries, programmes should focus on recipient-country priorities and absorptive capacity; translate research on global public goods into context-appropriate technologies; plan and monitor pathways to impact; structure equitable partnerships; strengthen individual and institutional capacity; and emphasize knowledge mobilization.

Conclusions
Global health and development research programmes and partnerships have an important role to play in achieving the SDGs and addressing global challenges. Governments should consider the potential of ODA-funded research programmes to address gaps in their global health and development frameworks. In the absence of concrete evidence of development impact, donor countries should consider making increases in ODA allocations for research additional to more direct investments that have demonstrated effectiveness in ODA-receiving countries.
Accelerometer-measured physical activity and sedentary time among children and their parents in the UK before and after COVID-19 lockdowns: a natural experiment
Authors: Salway Ruth , Foster Charlie , de Vocht Frank , Tibbitts Byron , Emm-Collison Lydia , House Danielle , Williams Joanna G. , Breheny Katie , Reid Tom , Walker Robert , Churchward Sarah , Hollingworth William , Jago Russell 
Published: May 2022

Abstract
Background
Restrictions due to the coronavirus disease 2019 (COVID-19) pandemic reduced physical activity provision for both children and their parents. Recent studies have reported decreases in physical activity levels during lockdown restrictions, but these were largely reliant on self-report methods, with data collected via unrepresentative self-report surveys. The post-pandemic impacts on children’s activity levels remain unknown. A key question is how active children become once lockdown restrictions are lifted.

Methods
Active-6 is a repeated cross-sectional natural experiment. Accelerometer data from 1296 children aged 10–11 and their parents were collected in 50 schools in the Greater Bristol area, UK in March 2017-May 2018 (pre-COVID-19 comparator group), and compared to 393 children aged 10–11 and parents in 23 of the same schools, collected in May-December 2021. Mean minutes of accelerometer-measured moderate-to-vigorous physical activity (MVPA) were derived for weekdays and weekend and compared pre- and post-lockdown via linear multilevel models.

Results
After adjusting for seasonality, accelerometer wear time and child/parent demographics, children’s mean weekday and weekend MVPA were 7.7 min (95% CI: 3.5 to 11.9) and 6.9 min (95% CI: 0.9 to 12.9) lower in 2021 than in 2018, respectively, while sedentary time was higher by 25.4 min (95% CI: 15.8 to 35.0) and 14.0 min (95% CI: 1.5 to 26.5). There was no evidence that differences varied by child gender or household education. There was no significant difference in parents’ MVPA or sedentary time, either on weekdays or weekends.

Conclusions
Children’s MVPA was lower by 7–8 min/day in 2021 once restrictions were lifted than before the pandemic for all groups, on both weekdays and weekends. Previous research has shown that there is an undesirable age-related decline in children’s physical activity. The 8-min difference reported here would be broadly comparable to the decline that would have previously been expected to occur over a three-year period. Parents’ physical activity was similar to pre-pandemic levels. Our results suggest that despite easing of restrictions, children’s activity levels have not returned to pre-pandemic levels. There is an urgent need to understand why these changes have occurred and how long they are maintained.
Interleukin 13 promotes long-term recovery after ischemic stroke by inhibiting the activation of STAT3
Authors: Chen Di , Li Jiaying , Huang Yichen , Wei Pengju , Miao Wanying , Yang Yaomei , Gao Yanqin 
Published: May 2022

AbstractBackgroundMicroglia/macrophages are activated after cerebral ischemic stroke and can contribute to either brain injury or recovery by polarizing microglia/macrophage into distinctive functional phenotypes with pro- or anti-inflammatory properties. Interleukin-13 (IL-13) is an anti-inflammatory cytokine that regulates microglia/macrophage polarization toward an anti-inflammatory phenotype. However, it is not clear whether IL-13 is beneficial after ischemic stroke long-term and the underlying molecular mechanism(s) remain unknown. Thus, we examined the effect of IL-13 on long-term recovery and microglia/macrophage polarization in mice with transient middle cerebral artery occlusion model (tMCAO).MethodstMCAO was induced in adult male C57BL/6J mice. IL-13 (60 μg/kg) was administered intranasally starting 2 h after stroke and continued for seven consecutive days. Sensorimotor function, spatial learning and memory function, as well as brain infarct volume were assessed up to 35 days after stroke. White matter integrity was evaluated by electrophysiology, immunofluorescence staining, and transmission electron microscopy. Microglia/macrophage activation was assessed using immunofluorescence staining and quantitative real-time polymerase chain reaction. Changes in immune cells in the brain and the periphery, and expression of IL-13 receptors in different brain cells were detected by flow cytometry. Primary neuron/microglia co-cultures and a STAT3 inhibitor were used for mechanistic studies.ResultsPost-treatment with IL-13 improved long-term neurofunctional recovery and decreased brain tissue atrophy after stroke. Intranasal delivery of IL-13 enhanced the structural and functional integrity of white matter after stroke. Furthermore, the neuroprotection afforded by IL-13 administration was not due to a direct effect on neurons, but by indirectly regulating the anti-inflammatory phenotype of microglia/macrophages. IL-13 treatment also had no effect on peripheral immune cells. Mechanistically, IL-13 improved the long-term outcome after ischemic stroke by promoting the polarization of microglia/macrophages toward the anti-inflammatory phenotype at least partially by inhibiting the phosphorylation of STAT3.ConclusionsIL-13 promotes white matter repair and improves neurofunctional outcomes after ischemic stroke by modulating microglia/macrophages via inhibition of STAT3 phosphorylation.
Manual therapy versus advice to stay active for nonspecific back and/or neck pain: a cost-effectiveness analysis
Authors: Aboagye Emmanuel , Lilje Stina , Bengtsson Camilla , Peterson Anna , Persson Ulf , Skillgate Eva 
Published: May 2022

AbstractBackgroundLow back and neck pain are the most common musculoskeletal disorders worldwide, and imply suffering and substantial societal costs, hence effective interventions are crucial. The aim of this study was to evaluate the cost-effectiveness of manual therapy compared with advice to stay active for working age persons with nonspecific back and/or neck pain.MethodsThe two interventions were: a maximum of 6 manual therapy sessions within 6 weeks, including spinal manipulation/mobilization, massage and stretching, performed by a naprapath (index group), respectively information from a physician on the importance to stay active and on how to cope with pain, according to evidence-based advice, at 2 occasions within 3 weeks (control group). A cost-effectiveness analysis with a societal perspective was performed alongside a randomized controlled trial including 409 persons followed for one year, in 2005. The outcomes were health-related Quality of Life (QoL) encoded from the SF-36 and pain intensity. Direct and indirect costs were calculated based on intervention and medication costs and sickness absence data. An incremental cost per health related QoL was calculated, and sensitivity analyses were performed.ResultsThe difference in QoL gains was 0.007 (95% CI − 0.010 to 0.023) and the mean improvement in pain intensity was 0.6 (95% CI 0.068–1.065) in favor of manual therapy after one year. Concerning the QoL outcome, the differences in mean cost per person was estimated at − 437 EUR (95% CI − 1302 to 371) and for the pain outcome the difference was − 635 EUR (95% CI − 1587 to 246) in favor of manual therapy. The results indicate that manual therapy achieves better outcomes at lower costs compared with advice to stay active. The sensitivity analyses were consistent with the main results.ConclusionsOur results indicate that manual therapy for nonspecific back and/or neck pain is slightly less costly and more beneficial than advice to stay active for this sample of working age persons. Since manual therapy treatment is at least as cost-effective as evidence-based advice from a physician, it may be recommended for neck and low back pain. Further health economic studies that may confirm those findings are warranted.Trial registrationCurrent Controlled Trials ISRCTN56954776. Retrospectively registered 12 September 2006,http://www.isrctn.com/ISRCTN56954776.
Safety of surgical hip dislocation in femoral head fracture and dislocation (FHFD) and avascular necrosis risk factor analysis of FHFD: midterm results confirmed by SPECT/CT and MRI
Authors: Yoon Yong-Cheol , Oh Chang-Wug , Kim Joon-Woo , Heo Jeong , Song Hyung Keun 
Published: May 2022

Abstract
Background
The study aim was to report the treatment outcomes of trochanteric flip osteotomy (TFO) with surgical hip dislocation (SHD) for femoral head fracture and dislocation (FHFD) and to investigate the risk factors for avascular necrosis (AVN) of the femoral head.

Methods
The data of 34 patients (29 men, 5 women; mean age 37.9 years) diagnosed with FHFD and treated with TFO with SHD between May 2009 and February 2018 with an average follow-up period of 5.1 years (range 2.8–10.5 years) were analyzed. Clinical outcomes were evaluated using the Merle d'Aubigné–Postel score and Thompson–Epstein Scale. Radiologic outcomes were classified according to the Matta classification. AVN was confirmed using magnetic resonance imaging or single-photon emission computed tomography/computed tomography. The occurrence of complications was examined, and factors influencing complications, AVN.

Results
Regarding the Pipkin’s classification, there were 7 patients with type II, 2 patients with type III, and 25 patients with type IV fractures. Posterior wall fractures accompanied all associated acetabular fractures in the patients with Pipkin type IV fractures. Radiologically, the union of acetabular and femoral head fractures was observed within 6.1 months on average (range 4–10 months) in 32 patients, except two patients who developed femoral head AVN. Clinically, the average Merle d'Aubigné–Postel score was 14.4 points (range 8–17 points), and 22 patients had good or excellent results on the Thompson–Epstein Scale. Two patients developed femoral head AVN with both having displaced femoral neck fractures associated with FHFD. AVN was significantly correlated with femoral neck fractures (P = 0.000).

Conclusion
TFO with SHD is a safe and useful approach for the treatment of FHFD. Particular attention should be paid when treating femoral head fractures associated with displaced femoral neck fractures because of the high risk of AVN development.
Quality of life following surgical repair of acute type A aortic dissection: a systematic review
Authors: Eranki Aditya , Wilson-Smith Ashley , Williams Michael L. , Saxena Akshat , Mejia Ross 
Published: May 2022

AbstractBackgroundThe outcomes of surgery for acute Stanford Type A aortic dissection (ATAAD) extend beyond mortality and morbidity. The aim of this systematic review was to summarise the literature surrounding health related quality of life (HR-QOL) following ATAAD, compare the outcomes to the standardised population, and to assess the impact of advanced age on HRQOL outcomes following surgery.MethodsA systematic review of studies after January 2000 was performed to identify HR-QOL in patients following surgery for ATAAD. Electronic searches of three databases were performed and clinical studies extracted by two independent reviewers. Strict inclusion and exclusion criteria were applied. Quality appraisal was conducted utilizing predefined criteria on pilot forms. HR-QOL results were synthesized through a narrative review of included studies.ResultsThere was significant attrition in HR-QOL of patients following surgery for ATAAD. Outcomes fared worse when compared to an age adjusted normative population. Of note, elderly patients were physically vulnerable, whereas younger populations may be more mentally vulnerable to postoperative sequalae. The included studies were quite heterogeneous in their study designs, methods, HR-QOL measures reported and follow up time-frames which limited direct comparison between studies.ConclusionHR-QOL outcomes are adversely affected when compared to preoperative status and physical health demonstrates significant attrition over time. HR-QOL outcomes are worse off when compared to an age matched general population. In terms of age, advancing age is associated with worse physical component scores but emotional health may fare better than younger patients.
A mixed method study on the impact of living with spinal muscular atrophy in Malaysia from patients’ and caregivers’ perspectives
Authors: Ch’ng Gaik Siew , Koh Karina , Ahmad-Annuar Azlina , Taib Fahisham , Koh Cha Ling , Lim Edmund Soon Chin 
Published: May 2022

Abstract
Background
Spinal Muscular Atrophy (SMA) is a rare, recessively inherited neuromuscular disorder that causes progressive muscle weakness. There is a low degree of awareness about SMA amongst the public and healthcare providers, which may impact the perception of the disease and its proper management. To understand how this lack of awareness may have affected diagnosis, care and support for SMA patients and their caregivers, this study aims to investigate the impact of SMA on the lives and daily activities of SMA patients and their caregivers in Malaysia.

Methods
Nationwide recruitment was carried out via invitations coordinated by a local SMA advocacy organization. A mixed method cross-sectional study consisting of a self-administered questionnaire followed by in-depth interviews (IDIs) and focus group discussions (FGDs) was conducted. The interview sessions were audio-taped, and verbatim transcripts analyzed thematically.

Results
Participants reported feeling stressed, anxious and depressed. There were issues with delayed diagnosis, lack of information from healthcare professionals about the disease progression, and limited access to supportive services like physiotherapy. Participants expressed their concerns living with self-doubt and turmoil with having to modify their lifestyles, relationships with family and friends, and challenges with educational and career opportunities. Various themes of their hopes for the future touched on having access to treatment, clinical trials, holistic care for symptom management, as well as improving infrastructure for disability access.

Conclusion
This study, to the best of our knowledge represents the first comprehensive study on SMA in South East Asia, highlights a plethora of issues and challenges experienced by persons with spinal muscular atrophy (PWSMA) and their caregivers in Malaysia, from the point of SMA diagnosis and throughout the management of care, in addition to the deep psychosocial impact of living with SMA. The significant findings of this study may contribute to a better understanding among stakeholders to make improvements in clinical practice, the education system, the work environment as well as holistic care support and society at large.