Experimental Animals Research Articles

Endovascular aortic occlusion improves return of spontaneous circulation after longer periods of cardiopulmonary resuscitation: A translational study in pigs

IntroductionResuscitative endovascular balloon occlusion of the aorta (REBOA) has emerged as an adjunct to CPR for nontraumatic cardiac arrest (NTCA). This translational study investigated the impact of varying low-flow duration (15- vs 30-mins) on REBOA's hemodynamic performance and ability to achieve return of spontaneous circulation (ROSC) in a porcine model. MethodsThirty-two pigs were anesthetized and placed into ventricular fibrillation. All animals received a 4-min no-flow period before CPR was initiated. Animals were randomized into four groups: 15- vs 30-minutes of CPR; REBOA vs. no-REBOA. After completion of 15- or 30-minute low-flow, ACLS was initiated and REBOA was inflated in experimental animals. ResultsIn the 15-mins groups, there were no differences in the rates of ROSC between REBOA (4/8, 50%) and control (4/8, 50%; p = 0.99). However, in the 30-min groups, the REBOA animals had a significantly higher rate of ROSC (6/8, 75%) compared to control (1/8, 12.5%; p = 0.04). In the 7-mins after REBOA deployment in the 30-min animals there was a statistically significant difference in coronary perfusion pressure (REBOA 42.1 mmHg, control 3.6 mmHg, p = 0.038). Importantly, 5/6 animals that obtained ROSC in the 30-min group with REBOA re-arrested at least once, with 3/6 maintaining ROSC until study completion. ConclusionIn our porcine model of NTCA, REBOA preferentially improved hemodynamics and ROSC after a 30-mins period of low-flow CPR. REBOA may be a viable strategy to improve ROSC after prolonged downtime, however, more hemodynamic support will be required to maintain ROSC.

Phosphorylation of cardiac sodium channel at Ser571 anticipates manifestations of the aging myopathy.

Diastolic dysfunction and delayed ventricular repolarization are typically observed in the elderly, but whether these defects are intimately associated with the progressive manifestation of the aging myopathy remains to be determined. In this regard, aging in experimental animals is coupled with increased late Na+ current (INa,L) in cardiomyocytes, raising the possibility that INa,L conditions the modality of electrical recovery and myocardial relaxation of the aged heart. For this purpose, aging male and female wild-type (WT) C57Bl/6 mice were studied together with genetically engineered mice with phosphomimetic (gain of function, GoF) or ablated (loss of function, LoF) mutations of the sodium channel Nav1.5 at Ser571 associated with, respectively, increased and stabilized INa,L. At ∼18 mo of age, WT mice developed prolonged duration of the QT interval of the electrocardiogram and impaired diastolic left ventricular (LV) filling, defects that were reversed by INa,L inhibition. Prolonged repolarization and impaired LV filling occurred prematurely in adult (∼5 mo) GoF mutant mice, whereas these alterations were largely attenuated in aging LoF mutant animals. Ca2+ transient decay and kinetics of myocyte shortening/relengthening were delayed in aged (∼24 mo) WT myocytes, with respect to adult cells. In contrast, delayed Ca2+ transients and contractile dynamics occurred at adult stage in GoF myocytes and further deteriorated in old age. Conversely, myocyte mechanics were minimally affected in aging LoF cells. Collectively, these results document that Nav1.5 phosphorylation at Ser571 and the late Na+ current modulate the modality of myocyte relaxation, constituting the mechanism linking delayed ventricular repolarization and diastolic dysfunction.NEW & NOTEWORTHY We have investigated the impact of the late Na current (INa,L) on cardiac and myocyte function with aging by using genetically engineered animals with enhanced or stabilized INa,L, due to phosphomimetic or phosphoablated mutations of Nav1.5. Our findings support the notion that phosphorylation of Nav1.5 at Ser571 prolongs myocardial repolarization and impairs diastolic function, contributing to the manifestations of the aging myopathy.

Efficacy of substances containing 3 types of active ingredients-saponins, flavones, and alkaloids in regulation of cytokines in autoimmune diseases a systematic review and Meta-analysis based on animal studies.

To investigate the efficacy of substances containing 3 types of active ingredients-saponins, flavones, and alkaloids on experimental animals with autoimmune diseases (AIDs). The protocol for this systematic review and Meta-analysis was prospectively registered with PROSPERO (CRD42023395741). Searches were conducted in the China National Knowledge Infrastructure, Wanfang, Chinese Science and Technology Journals, China Biomedical, PubMed, Cochrane Library, and Embase databases to screen for animal studies investigating the therapeutic effects of saponins, flavones, or alkaloids on autoimmune diseases; consequently, corresponding data extraction tables were prepared. Systematic Review Centre for Laboratory Animal Experimentation was used to assess the risk of methodological bias in the included literature. RevMan 5.4 was used for the Meta-analysis on the 8 serum cytokines. A total of 31 studies were included, all of which were randomized controlled studies. Meta-analysis indicated that substances rich in saponins, flavones, and alkaloids reduced serum levels of interleukin (IL)-1β [standardized mean difference (SMD) = -1.94, 95% confidence interval (CI) (-2.99, -0.90), P = 0.0003], IL-6 [SMD = -1.65, 95% CI (-2.33, -0.97,) P < 0.000 01], IL-17 [SMD = -2.41, 95% CI (-3.61, -1.20), P < 0.0001], tumor necrosis factor (TNF)-α [SMD = -1.84, 95% CI (-2.61, -1.06), P < 0.0001], and interferon (IFN)-γ [SMD = -1.54, 95% CI (-2.43, -0.65), P = 0.0007], but increased serum levels of IL-4 [SMD = 1.30, 95% CI (0.15, 2.44), P = 0.03) and IL-10 [SMD = 2.05, 95% CI (1.39, 2.70), P < 0.000 01) in animal models. However, no significant regulatory effect of these three active components was observed on serum levels of IL-2 [SMD = -0.63, 95% CI (-1.82, 0.57), P = 0.30]. Substances containing saponins, flavones, and alkaloids regulated the changes of immune-related cytokines, it may be a novel dietary substance to relieve and control autoimmune diseases in the future.

Network Pharmacology and Experimental Validation of Qingwen Baidu Decoction Therapeutic Potential in COVID-19-related Lung Injury

Background and Purpose: Coronavirus disease 2019 (COVID-19) is a lifethreatening disease worldwide due to its high infection and serious outcomes resulting from acute lung injury. Qingwen Baidu decoction (QBD), a well-known herbal prescription, has shown significant efficacy in patients with Coronavirus disease 2019. Hence, this study aims to uncover the molecular mechanism of QBD in treating COVID-19-related lung injury. Methods: Traditional Chinese Medicine Systems Pharmacology database (TCMSP), DrugBanks database, and Chinese Knowledge Infrastructure Project (CNKI) were used to retrieve the active ingredients of QBD. Drug and disease targets were collected using UniProt and Online Mendelian Inheritance in Man databases (OMIM). The core targets of QBD for pneumonia were analyzed by the Protein-Protein Interaction Network (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) to reveal the underlying molecular mechanisms. The analysis of key targets using molecular docking and animal experiments was also validated. Results: A compound-direct-acting target network mainly containing 171 compounds and 110 corresponding direct targets was constructed. The key targets included STAT3, c-JUN, TNF-α, MAPK3, MAPK1, FOS, PPARG, MAPK8, IFNG, NFκB1, etc. Moreover, 117 signaling pathways mainly involved in cytokine storm, inflammatory response, immune stress, oxidative stress and glucose metabolism were found by KEGG. The molecular docking results showed that the quercetin, alanine, and kaempferol in QBD demonstrated the strongest affinity to STAT3, c- JUN, and TNF-α. Experimental results displayed that QBD could effectively reduce the pathological damage to lung tissue by LPS and significantly alleviate the expression levels of the three key targets, thus playing a potential therapeutic role in COVID-19. Conclusion: QBD might be a promising therapeutic agent for COVID-19 via ameliorating STAT3-related signals.

Biological effects and mechanism of β-amyloid aggregation inhibition by penetrable recombinant human HspB5-ACD structural domain protein

Alzheimer's disease (AD) is a global medical challenge. Studies have shown that neurotoxicity caused by pathological aggregation of β-amyloid (Aβ) is an important factor leading to AD. Therefore, inhibiting the pathological aggregation of Aβ is the key to treating AD. The recombinant human HspB5-ACD structural domain protein (AHspB5) prepared by our group in the previous period has been shown to have anti-amyloid aggregation effects, but its inability to penetrate biological membranes has limited its development. In this study, we prepared a recombinant fusion protein (T-AHspB5) of TAT and AHspB5. In vitro experiments showed that T-AHspB5 inhibited the formation of Aβ1-42 protofibrils and had the ability to penetrate the blood-brain barrier; in cellular experiments, T-AHspB5 prevented Aβ1-42-induced oxidative stress damage, apoptosis, and inflammatory responses in neuronal cells, and its mechanism of action was related to microglia activation and mitochondria-dependent apoptotic pathway. In animal experiments, T-AHspB5 improved memory and cognitive dysfunction and inhibited pathological changes of AD in APP/PS1 mice. In conclusion, this paper is expected to reveal the intervention mechanism and biological effect of T-AHspB5 on pathological aggregation of Aβ1-42, provide a new pathway for the treatment of AD, and lay the foundation for the future development and application of T-AHspB5.

The Participation of Ca2+ Channels in Epilepsy: A Bibliometric Analysis of the Scientific Literature in Latin America.

Bibliometric analysis allows us to quantify and evaluate scientific activity, and it has become increasingly important in all areas of scientific literature. Thanks to these analyses, we can infer where science should put greater efforts into elucidating the underlying mechanisms of diseases that have yet to be fully described or investigated. This paper delves into published articles related to the involvement of calcium (Ca2+) channels in epilepsy, which is a condition with a high prevalence in Latin America. We followed the scientific publication on SCOPUS and analyzed the impact of publications from Latin America in the field of epilepsy and the study of Ca2+ channels. We identified the countries with the largest number of publications and found that 68% of them were experimental (animal models), while 32% were clinical. We also identified the main journals, growth over time, and citation numbers. We found a total of 226 works produced by Latin American countries from 1976 to 2022. The countries that have contributed the most to the topic are Brazil, Mexico, and Argentina, with occasional collaborations between them to make contributions to the study of epilepsy and Ca2+ channels. Additionally, we found that the journal with the most citations is Nature Genetics. The number of authors per article ranges from 1 to 242, and neuroscience journals are the preferred target for researchers, with a predilection for publishing original articles, although 26% of the publications are review articles.

A Review of Cartilage Defect Treatments Using Chitosan Hydrogels in Experimental Animal Models.

Chitosan (CS) is a polycationic polysaccharide comprising glucosamine and N-acetylglucosamine and constitutes a potential material for use in cartilage tissue engineering. Moreover, CS hydrogels are able to promote the expression of cartilage matrix components and reduce inflammatory and catabolic mediator production by chondrocytes. Although all the positive outcomes, no review has analyzed the effects of CS hydrogels on cartilage repair in animal models. This study aimed to review the literature to examine the effects of CS hydrogels on cartilage repair in experimental animal models. The search was done by the descriptors of the Medical Subject Headings (MeSH) defined below: "Chitosan," "hydrogel," "cartilage repair," and "in vivo." A total of 420 articles were retrieved from the databases Pubmed, Scopus, Embase, Lilacs, and Web of Science. After the eligibility analyses, this review reported 9 different papers from the beginning of 2002 through the middle of 2022. It was found that cartilage repair was improved with the treatment of CS hydrogel, especially the one enriched with cells. In addition, CS hydrogel produced an upregulation of genes and proteins that act in the cartilage repair process, improving the biomechanical properties of gait. In conclusion, CS hydrogels were able to stimulate tissue ingrowth and accelerate the process of cartilage repair in animal studies.

Exploring the multicomponent synergy mechanism of Zuogui Wan on postmenopausal osteoporosis by a systems pharmacology strategy.

To explore the multi-component synergistic mechanism of Zuogui Wan (, ZGW) in treating postmenopausal osteoporosis (PMOP). The main components and target genes of ZGW were screened via the Traditional Chinese Medicine Systems Pharmacology (TCMSP). In addition, the target gene sets of PMOP were derived from the GeneCards and Online Mendelian Inheritance in Man databases. The search tool for recurring instances of neighbouring genes (STRING) 11.0 software was used to analyze the interaction among intersecting genes. Cytoscape 3.6.1 software and the Matthews correlation coefficient (MCC) algorithm were used to screen the core genes. Fifty Sprague-Dawley female rats were randomly divided into the sham-operated (Sham) group and the four ovariectomized (OVX) subgroups. Rats subjected to Sham or OVX were administered with the vehicle (OVX, 1 mL water/100 g weight), 17β-estradiol (E2, 50 μg·kg-1·d-1), and lyophilized powder of ZGW at a low dose of 2.3 (ZGW-L) and high dose of 4.6 (ZGW-H) g·kg-1·d-1 for three months. The bone density and bone strength were assessed using dual-energy X-ray and three-point bending tests, respectively. Furthermore, enzyme-linked immun-osorbent assay, Hematoxylin-eosin staining, and western blot analysis were used to determine the potential pharmacological mechanisms of action of ZGW in PMOP. A total of 117 active compounds of ZGW were screened from the TCMSP. Furthermore, 108 intersecting genes of drugs and diseases were identified. Using STRING software and the MCC algorithm, ten core genes, including C-X-C chemokine living 8 (CXCL8), C-C chemokine receptor type 2 (CCR2), alpha-2a active receptor (ADRA2A), melatonin receptor type 1B (MTNR1B), and amyloid-beta A4 protein (APP), were identified. The anti-osteoporosis regulation network of ZGW was constructed using the Cytoscape software. The animal experiments demonstrated that ZGW groups significantly reduced the serum levels of β-C-terminal telopeptide of type I collagen (β-CTX) and increased serum levels of bone-specific alkaline phosphatase (BALP) (P < 0.05, P < 0.01). The OVX group exhibited a significant decrease in bone mineral density and bone strength compared with the Sham group (P < 0.01). Moreover, treatment with ZGW resulted in increased trabecular thickness, improved arrangement of trabecular structure, and reduced empty bone lacunae. Furthermore, treatment with ZGW significantly increased the protein expression of CXCL8, ADRA2A, and CCR2 (P < 0.05, P < 0.01), and significantly decreased the protein expression of Runx2 (P < 0.01). Furthermore, the ZGW and E2 groups demonstrated significantly increased BMD (P < 0.05, P < 0.01), improved bone strength (P < 0.05, P < 0.01), reduced expression of CXCL8, ADRA2A, and CCR2, and increased runt-related transcription factor 2 levels in bone tissue (P < 0.05, P < 0.01) compared with the OVX group. However, there were no significant differences in MTNR1B and APP expression among the groups. ZGW shows synergistic mechanisms in PMOP through multiple components, targets, and pathways.

Copper exposure induces inflammation and PANoptosis through the TLR4/NF-κB signaling pathway, leading to testicular damage and impaired spermatogenesis in Wilson disease

Copper is a toxic heavy metal that causes various damage when it accumulates in the body beyond the physiological threshold. Wilson disease (WD) is an inherited disorder characterized by impaired copper metabolism. Reproductive damage in male patients with WD is gradually attracting attention. However, the underlying mechanisms of copper toxicity are unclear. In this study, we investigated the role of inflammation and PANoptosis in testicular damage and impaired spermatogenesis caused by copper deposition using the WD model toxic milk (TX) mice. Copper chelator-penicillamine and toll-like receptor 4 (TLR4) inhibitor-eritoran were used to intervene in TX mice in our animal experiment methods. Testis samples were collected from mice for further analysis. The results showed that the morphology and ultrastructure of the testis and epididymis in TX mice were damaged, and the sperm counts decreased significantly. The TLR4/nuclear factor kappa-B (NF-κB) signaling pathway was activated by copper deposition, which led to the upregulation of serum and testicular inflammatory factors in TX mice. Meanwhile, pyroptosis, apoptosis, and necroptosis were significant in the testis of TX mice. Both chelated copper or inhibited TLR4 expression markedly suppressed the TLR4/NF-κB signaling pathway, thereby reducing the expression of inflammatory factors. PANoptosis in the testis of TX mice was also reversed. Our study indicated that pathological copper exposure induces inflammation and PANoptosis through the TLR4/NF-κB signaling pathway, leading to toxic testicular damage and impaired spermatogenesis in WD.

Oral microbiota dysbiosis alters chronic restraint stress-induced depression-like behaviors by modulating host metabolism

Studies have shown that the microbiota-gut-brain axis is highly correlated with the pathogenesis of depression in humans. However, whether independent oral microbiome that do not depend on gut microbes could affect the progression of depression in human beings remains unclear, neither does the presence and underlying mechanisms of the microbiota-oral-brain axis in the development of the condition. Hence this study that encompasses clinical and animal experiments aims at investigating the correlation between oral microbiota and the onset of depression via mediating the microbiota-oral-brain axis. We compared the oral microbial compositions and metabolomes of 87 patients with depressive symptoms versus 70 healthy controls. We found that the oral microbial and metabolic signatures were significantly different between the two groups. Significantly, germ-free (GF) mice transplanted with saliva from mice exposing to chronic restraint stress (CRS) displayed depression-like behavior and oral microbial dysbiosis. This was characterized by a significant differential abundance of bacterial species, including the enrichment of Pseudomonas, Pasteurellaceae, and Muribacter, as well as the depletion of Streptococcus. Metabolomic analysis showed the alternation of metabolites in the plasma of CRS-exposed GF mice, especially Eicosapentaenoic Acid. Furthermore, oral and gut barrier dysfunction caused by CRS-induced oral microbiota dysbiosis may be associated with increased blood-brain barrier permeability. Pseudomonas aeruginosa supplementation exacerbated depression-like behavior, while Eicosapentaenoic Acid treatment conferred protection against depression-like states in mice. These results suggest that oral microbiome and metabolic function dysbiosis may be relevant to the pathogenesis and pathophysiology of depression. The proposed microbiota-oral-brain axis provides a new way and targets for us to study the pathogenesis of depression.

Effect of ethanolic stem bark extract of Annona muricata (soursop) on reproductive hormones and ovary in female Wistar rats

The effect of ethanol extract of stem bark of Annona muricata (soursop) on reproductive hormones and ovaries of Wistar rats was studied. The phytochemical analysis revealed the presence of alkaloids, flavonoids, flavones, flavonols, xanthones, flavanonols, flavonones, and condensed tannins. Twenty (20) experimental animals were randomly grouped into five groups of 4 rats per group and were allowed to acclimatize for 2 weeks. Rats in group A served as the control and were fed with rat feed and water, rats in group B severed as positive control and received low dose clomiphene citrate (20 mg/kg) daily for 21 days, rat feed and water, rats in groups C to E were fed with normal rat feed, water and received different doses of fresh stem bark of Annona muricata extract at 250, 500 and 1000 mg/kg respectively for 21 days. The animals were sacrificed under anesthesia after the experiment and the ovaries were surgically extracted, weighed and sent for histology. Blood samples were collected for analysis (hormonal assay of luteinizing hormone (LH), estrogen, follicle stimulating hormone (FSH), progesterone and prolactin. This was carried out using enzyme linked immunosorbent assay (ELISA) technique. The results showed that there were significant changes in the hormone level of the test groups when compared with the control rats. Significant increase was recorded in progesterone, estrogen, prolactin, follicle stimulating hormone and luteinizing hormone p&lt; 0.05. Hence Annona muricata improves fertility in female Wistar rats. There were no histological changes in the ovaries and hence, it was concluded that Annona Muricata does not cause organ damage with increasing dose.

Evaluation of anti- inflammatory and analgesic activity of Bhasmas in experimental animal models

Acute toxicity studies of both the bhasmas showed no mortality at 2000 mg/kg.The Scanning electron microscopy of both Tamra and Lauha bhasma indicated the nanomaterial characteristics which justifies claim of Ayurvedic bhasmas as ancient nanomaterials. Tamra and Lauha bhasma were found to be effective as anti-inflammatory agent in acute models of inflammation. It inhibits all phases of inflammation induced by carrageenan. Tamra and Lauha bhasma were found to be effective as anti-inflammatory agent in chronic model of inflammation. It significantly inhibits wet as well as dry weight of cotton pellet granuloma at highest doses only. Analgesic activity of both Tamra and Lauha bhasma in hot plate and tail immersion models were found to be significant at 120-240 mins. Analgesic activity of both Tamra and Lauha bhasma in acetic acid models were found to be non-significant in nature, while found to be significant at late phase in formalin induced model. Tamra and Lauha bhasma also were found to have significant anti-arthritic activity in animal study.The whole pharmacological investigations were indicated that activity was only significant at therapeutic and more than therapeutic dose but nonsignificant at dose lower than therapeutic doses. The obtained results indicates the anti-inflammatory and analgesic potential of both these bhasmas probably by inhibiting inflammatory mediators.

Mechanistic analyses reveal that Pueraria montana var. lobata (Willd.) is effective in inhibiting ovarian cancer progression.

Ovarian cancer (OC) is a common malignancies of the female genitalia. P. montana var. lobata (Willd.), a herb with anti-tumor effects, is widely used in the clinical treatment of ovarian cancer (OC), but the ingredients and molecular mechanism of action remains to be explored. In this study, we extracted the main active ingredients of P. montana var. lobata (Willd.) from the TCMSP database, and predicted its potential targets of action against OC from the DisGeNET and GeneCards databases. Protein-protein interaction (PPI) was constructed using the STRING database, while pathway enrichment analyses were performed using the DAVID database. Next, we generated an Ingredient-Target-Pathway network using Cytoscape 3.7.2, then processed the key targets of action and main active ingredients for molecular docking.The results showed that seven active ingredients of P montana var. lobata (Willd.) were associated with treating for OC, namely beta-sitosterol, coumestrol, daidzein, formononetin, genistein, puerarin and scoparone, two important targets Casp3 and Jun, and signaling pathways of P. montana var. lobata (Willd.) against the progression of OC. TUNEL staining, enzyme-linked immunosorbent assay (ELISA), and Western blot assays, the pharmacodynamic effect of puerarin in the treatment of OC and the major targets were verified. Animal experiment demonstrated that application of puerarin at different times of modeling not only upregulated expression of Casp3, Smac, and c-jun proteins, but also promoted apoptosis in tumor cells, hence inhibiting progression of OC. This study demonstrates that P. montana var. lobata (Willd.) can thereby induce apoptosis in tumor cells and inhibit malignant progression through activating expression of Casp3, smac, and c-jun proteins to regulate related apoptosis pathways, as validated by network pharmacology predictions and animal experiments, and can be verifed by large-scale clinical trials in the future. This study also provides theoretical support and new research perspectives for this disease.

A Review on Techniques of Blood Withdrawal in Experimental Small Animals

In preclinical research, experimental studies in small animals like pharmacokinetics/ toxicokinetics, serum biochemistry and hematology necessitate the biological matrix sampling to estimate drug level, serum enzyme, and blood parameters respectively. The biological matrix is withdrawn from experimental animals by employing the recommended blood collection technique and must be conducted in compliance with the ethical guidelines. It has demanded a minimal painful and least stressful procedures, which can be achieved using inhalant/parenteral anesthesia and/or proper restraint technique. This leads to better experimental outcome. The study personnel conducting blood withdrawal must be well-versed. All these procedures must be approved by IAEC protocol and should include scientist and study personnel names who are involved in animal handling and blood withdrawal. In this review, the different techniques of blood withdrawal from small animals, including collection guidelines, limit volumes and recovery periods, and advantages and disadvantages of sites used for blood collection are demonstrated and discussed.

Zinc and Inflammatory Bowel Disease: From Clinical Study to Animal Experiment.

Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gastrointestinal tract (GI) with a high incidence rate globally, and IBD patients are often accompanied by zinc deficiency. This review aims to summarize the potential therapeutic value of zinc supplementation in IBD clinical patients and animal models. Zinc supplementation can relieve the severity of IBD especially in patients with zinc deficiency. The clinical severity of IBD were mainly evaluated through some scoring methods involving clinical performance, endoscopic observation, blood biochemistry, and pathologic biopsy. Through conducting animal experiments, it has been found that zinc plays an important role in alleviating clinical symptoms and improving pathological lesions. In both clinical observation and animal experiment of IBD, the therapeutic mechanisms of zinc interventions have been found to be related to immunomodulation, intestinal epithelial repair, and gut microbiota's balance. Furthermore, the antioxidant activity of zinc was clarified in animal experiment. Appropriate zinc supplementation is beneficial for IBD therapy, and the present evidence highlights that alleviating zinc-deficient status can effectively improve the severity of clinical symptoms in IBD patients and animal models.

How to optimize communication about animal and animal-free research methods

There is a long-standing debate about experimental non-human animals and animal-free methods in scientific research. Among the various stakeholders involved in the debate are the scientists. During media broadcasts we, animal researchers and animal-free methods researchers, were positioned as ‘opponents’. In this essay we describe our initial rational thoughts and emotions after these events, and how we came together to explore our common ground on animal(−free) experimentation. Realizing that all models have advantages and limitations, our common ground lies in the principles of good scientific research and responsible experimentation. Our communication emanating from the broadcasts has been instrumental in improving communication on animal(−free) experimentation issues by teaming up. We strongly believe that this is essential for making well-informed decisions for the methods we are using now and will be using in the future.

Lijing-Anfei Decoction Attenuates COPD in a Mouse Model: Network Pharmacology Analysis, Molecular Docking, and Experimental Validation

Background Today, network pharmacology and molecular docking were utilized for investigation and prediction of possible mechanisms in fields such as pharmacognosy and traditional Chinese medicine formulas when treating disease. Purpose Chronic obstructive pulmonary disease (COPD) is caused by exposure to toxic particles and leads to chronic bronchitis and lung dysfunction. Lijing-Anfei decoction (LAD) is a traditional Chinese herbal decoction used to cure COPD. This study explored the effects of this decoction and its intrinsic mechanism in a COPD mouse model. Materials and methods Possible targets were retrieved from several databases and combined using Veny. Protein-protein interaction (PPI) networks were constructed via STRING database. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed via the Metascape database. The COPD model was established by exposing the mice to cigarette smoke for 12 weeks. After the model was established, LAD was administered for four weeks. Enzyme-linked immunosorbent assay (ELISA) was performed, and body weights and lung wet/dry ratios were measured. Lung morphology was observed by hematoxylin-eosin staining. Results After the screening, 32 overlapping targets were used to generate PPI networks and the following hub genes were identified: JUN, IL6, RELA, IL1B, HIF1A, MAPK1, PPARG, PTGS2, MAPK14, and NOS2. The KEGG analysis indicated that hypoxia-inducible factor-1 (HIF-1) and other signaling pathways are possible signaling pathways. The molecular docking results demonstrated tight links between major compounds and their targets. In animal experiments, LAD attenuated body weight loss, downregulated inflammatory cell numbers, and inhibited the enhancement of Fe2+ levels. The mean linear intercept index changes indicated that LAD ameliorated the lung structural damage. ELISA revealed that it downregulated pro-inflammatory cytokine levels and inhibited excessive oxidant production, TdT-mediated dUTP-biotin nick end labeling assay demonstrated that it alleviated cell death in the lungs, and immunohistochemical staining revealed that it inhibited HIF-1α and PTGS2. ML228—an agonist of HIF-1—blocked the effect of LAD, validating the role of HIF-1-mediated ferroptosis in the pharmacological effects of this decoction. Conclusion This study revealed that LAD was successful in treating COPD.

LncRNA PART1 promotes malignant biological behaviours associated with head and neck cancer cells via synergistic action with FUT6

The aim of this study was to determine the role of lncRNA PART1 and downstream FUT6 in tumorigenesis and progression of head and neck cancer (HNC). Bioinformatics analysis and qRT-PCR revealed that lncRNA PART1 was expressed at low levels in HNC patients. The proliferation, apoptosis, migration and flow cytometry results showed that low expression of lncRNA PART1 inhibited apoptosis and promoted HNC cell migration and proliferation. In addition, animal experiments have also shown that low expression of lncRNA PART1 can promote tumor growth. LncRNA PART1 overexpression promoted apoptosis and inhibited HNC cell migration and proliferation. Through bioinformatics analysis, FUT6 was found to be expressed at low levels in HNC and to be correlated with patient survival. Immunohistochemical and qRT-PCR results revealed that FUT6 was underexpressed in tumour tissues and HNC cells. Cell and animal experiments showed that overexpression of FUT6 could inhibit tumour proliferation and migration. Bioinformatics analysis revealed that lncRNA PART1 was positively correlated with FUT6. By qRT-PCR and western blot, we observed that after knockdown of lncRNA PART1, both the mRNA and protein expression levels of FUT6 were reduced. The above results indicated that lncRNA PART1 and FUT6 play an important role in HNC, and that lncRNA PART1 affected the development of tumor by downstream FUT6.

Characterization and Efficacy of C60 Nano-Photosensitive Drugs in Colorectal Cancer Treatment

BackgroundFullerenes C60 shows great potential for drug transport. C60 generates large amounts of singlet oxygen upon photoexcitation, which has a significant inhibitory effect on tumor cells, so the photosensitive properties of C60 were exploited for photodynamic therapy of tumors by laser irradiation. MethodsIn this study, C60-NH2 was functionalized by introducing amino acids on the surface of C60, coupled with 5-FU to obtain C60 amino acid-derived drugs (C60AF, C60GF, C60LF), and activated photosensitive drugs (C60AFL, C60GFL, C60LFL) were obtained by laser irradiation. The C60 nano-photosensitive drugs were characterized in various ways, and the efficacy and safety of C60 nano-photosensitive drugs were verified by cellular experiments and animal experiments. Bioinformatics methods and cellular experiments were used to confirm the photosensitive drug targets and verify the therapeutic targets with C60AF. ResultsPhotosensitised tumor-targeted drug delivery effectively crosses cell membranes, leads to more apoptotic cell death, and provides higher anti-tumor efficacy and safety in vitro and in vivo colorectal cancer pharmacodynamic assays compared to free 5-FU.C60 photosensitized drug promotes tumor killing by inhibiting the colorectal cancer FLOR1 tumor protein target, with no significant toxic effects on normal organs. ConclusionC60 photosensitized drug delivery systems are expected to improve efficacy and reduce side effects in the future treatment of colorectal cancer. Further and better development and design of drugs and vectors for colorectal cancer therapy.

Development of an ICT Laparoscopy System with Motion-Tracking Technology for Solo Laparoscopic Surgery: A Feasibility Study

The increasing demand for laparoscopic surgery due to its cosmetic benefits and rapid post-surgery recovery is juxtaposed with a shortage of surgical support staff. This juxtaposition highlights the necessity for improved camera management in laparoscopic procedures, encompassing positioning, zooming, and focusing. Our feasibility study introduces the information and communications technology (ICT) laparoscopy system designed to aid solo laparoscopic surgery. This system tracks a surgeon’s body motion using a controller, manipulating an embedded camera to focus on specific surgical areas. It comprises a camera module, a camera movement controller, and a motor within the main body, operating connected wires according to controller commands for camera movement. Surgeon movements are detected by an inertial measurement unit (IMU) sensor, facilitating precise camera control. Additional features include a foot pedal switch for motion tracking, a dedicated trocar for main body stability, and a display module. The system’s effectiveness was evaluated using an abdomen phantom model and animal experimentation with a porcine model. The camera responded to human movement within 100 ms, a delay that does not significantly affect procedural performance. The ICT laparoscopy system with advanced motion-tracking technology is a promising tool for solo laparoscopic surgery, potentially improving surgical outcomes and overcoming staff shortages.