Animal Models and Experimental Medicine

Background Fuke Huahuang formulation (FHF) is widely used in the treatment of vaginitis, with clinical evidence indicating its promising anti‐inflammatory properties. Methods We explored the bioactive components and potential mechanisms of FHF for treating vaginitis, and reveal its pharmacological activities against vaginitis. Results A total of 12 anti‐inflammatory components in FHF and 584 pharmacological targets were identified. Furthermore, 1427 vaginitis‐associated targets were identified, and 184 intersection targets between FHF and vaginitis were constructed for network analysis. Gene Ontology and pathway analysis revealed that the therapeutical targets of FHF against vaginitis are involved in modulating inflammatory stress, enhancing immunoregulation, reconstructing the microenvironment, and suppressing cell damage. Molecular docking analysis further suggested the possible direct binding of the bioactive compounds of FHF (fumarine) to the core targets, including AKT Serine/Threonine Kinase 1 (AKT1), Signal Transducer and Activator of Transcription 3 (STAT3), and nuclear factor‐kappaB (NF‐κB). Experimental validation found that FHF‐treated vaginitis rats exhibited reduced intracellular AKT1, STAT3, and NF‐κB protein expressions. Conclusion Overall, we identified the bioactive compounds and pharmacological mechanisms of FHF against vaginitis, thus offering a theoretical fundament for exploring FHF for treating vaginitis in the future.

Background Intracerebral hemorrhage (ICH) remains a devastating neurological disorder with limited therapeutic options. Neural stem cell (NSC)‐based therapies have emerged as a potential regenerative approach, yet the molecular mechanisms regulating NSC behavior require further elucidation. The role of miR‐21 in NSC differentiation and proliferation during ICH recovery remains unexplored. Methods In vitro NSC cultures were analyzed for miR‐21 expression dynamics during differentiation via qPCR. Lentiviral overexpression and knockdown of miR‐21 were employed to assess its functional impact. The SOX2/LIN28‐let‐7 pathway was investigated using Western blot, luciferase reporter assays, and immunofluorescence. In vivo, miR‐21‐overexpressing NSCs were transplanted into a murine ICH model, with neurogenesis evaluated by immunostaining and neurological recovery assessed through behavioral tests (mNSS, rotarod). Results miR‐21 expression significantly increased during NSC differentiation, correlating with reduced SOX2 levels. Mechanistically, miR‐21 directly targeted SOX2, disrupting the SOX2/LIN28‐let‐7 axis to promote NSC proliferation and lineage commitment. In ICH mice, transplantation of miR‐21‐overexpressing NSCs enhanced neurogenesis and improved motor coordination and neurological deficits at 28 days post‐transplantation. Conclusions Our findings identify miR‐21 as a critical regulator of NSC plasticity through SOX2/LIN28‐let‐7 signaling, highlighting its therapeutic potential for enhancing neuroregeneration and functional recovery in ICH. Targeting miR‐21 may represent a novel strategy to optimize NSC‐based therapies for hemorrhagic stroke.

Sydenham chorea (SC) is the neurological manifestation associated with acute rheumatic fever (ARF). ARF and rheumatic heart disease (RHD) are autoimmune complications triggered by a group A streptococcal (GAS) infection. In ARF/RHD and SC, tissue cross‐reactive antibodies and T‐cells generated against GAS antigens have been implicated in the pathogenesis. In SC, antibodies against GAS antigens are known to cross‐react with neuronal proteins causing neurological manifestations including choreiform movements and neuropsychiatric symptoms such as irritability, attention deficit, and obsessive‐compulsive disorder. Previous studies in a rat autoimmune valvulitis (RAV) model of RHD, have shown that injection of streptococcal M protein could cause both cardiac and neurological symptoms. In this study it was shown that adoptive transfer of serum with anti‐GAS M antibodies to naive rats caused carditis but failed to demonstrate neurobehavioral symptoms. However, when the blood–brain barrier (BBB) was disrupted using lipopolysaccharide, all animals that received anti‐GAS M protein antibodies, developed neurobehavioral defects in addition to carditis. This highlights that impaired BBB integrity is essential for the development of neurobehavioral symptoms. The use of the RAV model and the disruption of BBB required for the development of neurobehavioral changes provides a platform to further investigate the mechanisms that lead to antibodies binding to basal ganglia structures that cause SC.

Background Dry eye disease (DED) predominantly results from elevated tear film osmolarity, which can not only cause ocular inconvenience but may lead to visual impairments, severely compromising patient well‐being and exerting substantial economic burdens as well. Astaxanthin (AST), a member of the xanthophylls and recognized for its robust abilities to combat inflammation and oxidation, is a common dietary supplement. Nonetheless, the precise molecular pathways through which AST influences DED are still poorly understood. Methods Therapeutic targets for AST were identified using data from the GeneCards, PharmMapper, and Swiss Target Prediction databases, and STITCH datasets. Similarly, targets for dry eye disease (DED) were delineated leveraging resources such as the Therapeutic Target Database (TTD), DisGeNET, GeneCards, and OMIM databases, and DrugBank datasets. Interactions among shared targets were charted and displayed using CytoScape 3.9.0. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were conducted to elucidate the functions of pivotal targets within the protein–protein interaction network. Molecular interactions between AST and key targets were confirmed through molecular docking using AutoDock and PyMOL. Molecular dynamics simulations were performed using GROMACS 2022.3. Viability of human corneal epithelial cells (hCEC) was assessed across varying concentrations of AST. A mouse model of experimental DED was developed using 0.1% benzalkonium chloride (BAC), and the animals were administered 100 mg/kg/day of AST orally for 7 days. The efficacy of the treatments was assessed through a series of diagnostic tests to evaluate the condition of the ocular surface after the interventions. The levels of inflammation and oxidative stress were quantitatively assessed using methods such as reverse transcription‐polymerase chain reaction (RT‐PCR), Western blot, and immunofluorescence staining. Results Network pharmacology suggests that AST may alleviate DED by influencing oxidation–reduction signaling pathways and reducing oxidative stress provoked by BAC. In vivo experiments demonstrated an improved overall condition in AST‐administered mice in contrast to the control group. Immunofluorescence staining analyses indicated a decrease in Keap1 protein in the corneal tissues of AST‐treated mice and a significant increase in Nrf2 and HO‐1 protein. In vitro studies demonstrated that AST significantly enhanced cell viability and suppressed reactive oxygen species expression under hyperosmotic (HS) conditions, thereby protecting the human corneal epithelium. Conclusion AST is capable of shielding mice from BAC‐induced DED, decelerating the progression of DED, and mitigating oxidative stress damage under HS conditions in hCEC cells. The protective impact of AST on DED may operate through stimulating the Keap1‐Nrf2/HO‐1 signaling pathway. Our research findings indicate that AST may be a promising treatment for DED, offering new insights into DED treatment.

The World Health Organization has declared that COVID‐19 no longer constitutes a “public health emergency of international concern,” yet the long‐term impact of SARS‐CoV‐2 infection on bone health continues to pose new challenges for global public health. In recent years, numerous animal model and clinical studies have revealed that severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection can lead to secondary osteoporosis. The mechanisms involved are related to the virus's direct effects on bone tissue, dysregulation of the body's inflammatory response, hypoxia, noncoding RNA imbalance, and metabolic abnormalities. Although these studies have unveiled the connection between SARS‐CoV‐2 infection and osteoporosis, current research is not comprehensive and in depth. Future studies are needed to evaluate the long‐term effects of SARS‐CoV‐2 on bone density and metabolism, elucidate the specific mechanisms of pathogenesis, and explore potential interventions. This review aims to collate existing research literature on SARS‐CoV‐2 infection‐induced secondary osteoporosis, summarize the underlying mechanisms, and provide direction for future research.

Chinese hamster with Chinese characteristics is used in experiments, and it is of great value in the field of medical biology research. However, at present, there is no high‐efficiency method for evaluating the genetic quality of Chinese hamsters. Here, we developed a novel Chinese hamster genetic quality detection system using single‐nucleotide polymorphism (SNP) markers. To find SNP loci, we conducted whole genome sequencing on 24 Chinese hamsters. Then, we employed an SNP locus screening criterion that we set up previously and initially screened 214 SNP loci with wide genome distribution and high polymorphism level. Subsequently, we developed the SNP detection system using a multitarget region capture technique based on second‐generation sequencing, and a 55 SNP panel for genetic evaluation of Chinese hamster populations was developed. PopGen.32. analysis results showed that the average effective allele number, Shannon index, observed heterozygosity, expected heterozygosity, average heterozygosity, polymorphism information, and other genetic parameters of Chinese hamster population A were higher than those in population B. Using scientific screening and optimization, we successfully developed a novel Chinese hamster SNP genetic detection system that can efficiently and accurately analyze the genetic quality of the Chinese hamster population.

Background Due to the widespread use of cell phone devices today, numerous research studies have focused on the adverse effects of electromagnetic radiation on human neuropsychological and reproductive systems. In most studies, oxidative stress has been identified as the primary pathophysiological mechanism underlying the harmful effects of electromagnetic waves. This paper aims to provide a holistic review of the protective effects of melatonin against cell phone‐induced electromagnetic waves on various organs. Methods This study is a systematic review of articles chosen by searching Google Scholar, PubMed, Embase, Scopus, Web of Science, and Science Direct using the keywords ‘melatonin’, ‘cell phone radiation’, and ‘animal model’. The search focused on articles written in English, which were reviewed and evaluated. The PRISMA process was used to review the articles chosen for the study, and the JBI checklist was used to check the quality of the reviewed articles. Results In the final review of 11 valid quality‐checked articles, the effects of melatonin in the intervention group, the effects of electromagnetic waves in the case group, and the amount of melatonin in the chosen organ, i.e. brain, skin, eyes, testis and the kidney were thoroughly examined. The review showed that electromagnetic waves increase cellular anti‐oxidative activity in different tissues such as the brain, the skin, the eyes, the testis, and the kidneys. Melatonin can considerably augment the anti‐oxidative system of cells and protect tissues; these measurements were significantly increased in control groups. Electromagnetic waves can induce tissue atrophy and cell death in various organs including the brain and the skin and this effect was highly decreased by melatonin. Conclusion Our review confirms that melatonin effectively protects the organs of animal models against electromagnetic waves. In light of this conclusion and the current world‐wide use of melatonin, future studies should advance to the stages of human clinical trials. We also recommend that more research in the field of melatonin physiology is conducted in order to protect exposed cells from dying and that melatonin should be considered as a pharmaceutical option for treating the complications resulting from electromagnetic waves in humans.

The graphical abstract of this review highlights the critical role of humanized mice in the safety evaluation of antibody drugs. First, the inherent immunogenicity of antibody drugs can lead to the production of antidrug antibodies and hypersensitivity reactions on repeated administration. Therefore, it is essential to use transgenic animal models to assess the preclinical safety of antibody drugs at preclinical evaluation, which helps identify potential safety issues related to humanized antibodies. This article focuses on human immune system mice and transgenic humanized mice, discussing their applications in drug safety evaluation, including cytokine release induced by drugs and the assessment of drug side effects. However, humanized animal models face challenges such as graft‐versus‐host disease and poor reconstitution of human immune cells. Additionally, this article compares the differences between hIgG1 transgenic mice, human FcRn (hFcRn) transgenic mouse models, and wild‐type mice in the evaluation of antibody drugs. Despite the broad prospects of humanized animal models, there are still obstacles. With the advancement of induced pluripotent stem cell (iPSC) technology, these models are expected to be further improved. In the future, the development of various humanized immune checkpoint models will facilitate the discovery of antibody drugs, research on combination therapies, and guidance for personalized immunotherapy. image

Background Developing a granulomatous liver injury preclinical model may pave the way to understanding hepatic‐TB (tuberculosis) and autoimmune granulomatous liver diseases. Antitubercular (ATT) and other drugs' metabolism in the presence of a specific type of liver injury is not well understood. The present study aimed to establish a preclinical model of granulomatous hepatitis by using the BCG (Bacillus Calmette‐Guérin) vaccine, further studying it in the presence of ATT dosing, and analyze the pharmacokinetics of isoniazid, rifampicin, and their respective primary metabolites. Methods We used 56 rats in seven equal groups. Group I functioned as a normal control (NC) receiving normal saline only. Groups II–IV received intravenous injections of low‐, medium‐, and high‐dose BCG vaccine daily for 21 days. Groups V, VI, and VII received isoniazid (H) alone, rifampicin (R) alone, and isoniazid + rifampicin(HR) for a subsequent 15 days in addition to high dose BCG for the first 21 days, respectively. Liver function tests (LFT) were monitored on days 0, 21, 28, and 36. Rats were sacrificed later for oxidative stress and histopathological examination. Results The study observed BCG dose‐specific LFT derangements in groups II–IV compared to group I on day 21 (p < 0.05). Isoniazid, rifampicin, and combination intervention groups demonstrated normalization of the BCG‐led LFT changes. Histology and oxidative stress parameters confirmed model development and biochemical changes. Isoniazid area under the curve (AUC) showed a reduction of 16.9% in BCG + HR group in comparison to the BCG + H group (p = 0.01). Des‐acetyl‐rifampicin AUC and maximum‐concentration value demonstrated a significant rise in BCG + HR group in comparison to the BCG + R group (p = 0.001). Conclusion A novel preclinical model of granulomatous liver injury was developed using the BCG vaccine strain and validated with ATT response.

Background Cannabidiol (CBD) has numerous therapeutic properties, and is used to treat neurological conditions, such as neuroinflammation. However, the optimal dose of CBD to penetrate the brain requires further investigation. The primary aim of this study was to use a mouse model and the intrabuccal route for CBD administration to determine the optimal dose at which CBD can penetrate the brain. The secondary aim was to determine whether sex is a confounding factor. Methods Thirty adult Kramnik mice, divided equally into three groups, were administered CBD oil intrabuccally at three doses—10, 20, and 30 mg/kg, euthanized 6 h later, and whole brain, urine, and blood samples were collected. Liquid chromatography with tandem mass spectrometry was used to analyze the collected samples. Results CBD and its three metabolites—7‐carboxy cannabidiol (7‐COOH‐CBD), 7‐hydroxy cannabidiol (7‐OH‐CBD) and 6‐hydroxy cannabidiol (6‐OH‐CBD), were identified and quantified in all samples. The 10 and 20 mg/kg doses of CBD produced similar results in the brain, but the group given the 10 mg/kg dose had the least variation. The 30 mg/kg dose yielded the highest abundance of CBD and its metabolites in all samples, but also the greatest variation. Sex only became a confounding factor at 30 mg/kg. Conclusions This study shows that the intrabuccal route of CBD administration is reliable and the 10 mg/kg dose of CBD is recommended in mice because there were good CBD metabolite concentrations in all samples, with the least variation among the doses, and sex was not a confounder at 10 mg/kg.

Adenomatous polyposis coli (APC) mutations are the most frequently identified genetic alteration in sporadic colorectal cancer (CRC) cases, and a myriad of genetically engineered Apc‐mutant CRC mouse models have been developed using various genetic manipulation techniques. The advent of the CRISPR/Cas9 system has revolutionized the field of genetic engineering and facilitated the development of new genetically engineered mouse models. In this study, we aimed to develop a novel Apc knockout allele using the CRISPR/Cas9 system and evaluate the phenotypic effects of this new allele in two different mouse strains. For this purpose, exon 16 of mouse Apc gene was targeted with a single‐guide RNA, and the mouse carrying an Apc frameshift mutation at codon 750 (Δ750) was chosen as the founder. The mutant FVB‐ApcΔ750 mice were backcrossed with wild‐type C57BL/6 mice, and the phenotypic effects of the knockout allele were evaluated in F8‐FVB‐ApcΔ750, F4‐B6;FVB‐ApcΔ750, and F1‐B6;FVB‐ApcΔ750 by a macroscopic and microscopic examination of the gastrointestinal system. The result showed that the mean polyp number was significantly higher in F4‐BL6;FVB‐ApcΔ750 than in F8‐FVB‐ApcΔ750. Intestinal polyposis was more prominent in F4‐BL6;FVB‐ApcΔ750, whereas a higher number of colon polyps than intestinal polyps were observed in F8‐FVB‐ApcΔ750. Additionally, F1‐BL6;FVB‐ApcΔ750 mixed background mice developed gastric polyps that morphologically resembled the pyloric gland adenoma of humans. In conclusion, we developed a novel CRISPR‐mediated Apc knockout allele using two mouse strains. We showed that this allele can exert a strain‐specific effect on the phenotype of mice and can cause gastric polyp formation.

Background Refined models of kidney disease are critical to better understand disease processes and study novel treatments while minimizing discomfort in research animals. The objective of this study was to report a technique for minimally invasive partial kidney embolism in cats and describe outcomes following transcatheter administration of embolic microspheres with subsequent contralateral nephrectomy. Methods Eleven, apparently healthy, male, purpose‐bred cats underwent unilateral kidney embolism with 0.25 or 0.5 mL of embolic microparticle (40–120 μm) suspension (0.2 mL microspheres/mL) delivered into the right renal artery under fluoroscopic guidance, followed 5 months later by contralateral nephrectomy. One month after nephrectomy, blood and urinary markers of kidney function were evaluated, and embolized kidneys were harvested for histopathology evaluation. Results Renal artery embolization was possible in all cats. Two cats did not complete the study, one after experiencing congestive heart failure (n = 1) and the other following evidence of complete kidney embolism precluding nephrectomy (n = 1) post‐embolization. At study end, compared to baseline, cats had significant increases in median (range) serum creatinine (159.1 μmol/L [141.4–530.4] versus 128.2 μmol/L [92.8–150.3]; p = 0.0004), urea nitrogen (15.71 mmol/L [9.29–47.85] versus 7.50 mmol/L [6.07–8.57]; p < 0.0001), and symmetric dimethylarginine (0.74 μmol/L [0.59–3.12] versus 0.67 μmol/L [0.54–0.72]; p = 0.0288) concentrations. No differences in markers of kidney function were documented between dose groups. Conclusions Minimally invasive kidney embolism is a promising technique for modeling kidney disease in cats. Understanding optimal dose, timing of nephrectomy, and longer‐term consequences requires additional work.

Background Over the past 50 years, the incidence of obesity has gradually increased, necessitating investigation into the multifactorial contributors to this disease, including the gut microbiota. Bacteria within the human gut microbiome communicate using a density‐dependent process known as quorum sensing (QS), in which autoinducer (AI) molecules (e.g., N‐acyl‐homoserine lactones [AHLs]) are produced to enable bacterial interactions and regulate gene expression. Methods We aimed to disrupt QS using quorum quenching (QQ) lactonases GcL and SsoPox, which cleave AHL signaling molecules in a taxa‐specific manner based on differing enzyme affinities for different substrates. We hypothesized that QQ hinders signals from obesity‐associated pathobionts, thereby slowing or preventing obesity. Results In a murine model of diet‐induced obesity, we observed GcL and SsoPox treatments have separate sex‐dependent and dose‐dependent effects on intestinal community composition and diversity. Notably, male mice given 2 mg/mL SsoPox exhibited significant changes in the relative abundances of gram‐negative taxa, including Porphyromonadaceae, Akkermansiaceae, Muribaculaceae, and Bacteroidales (Kruskal–Wallis p < 0.001). Additionally, we used covariance matrix network analysis to model bacterial taxa co‐occurrence due to QQ enzyme administration. There were more associations among taxa in control mice, particularly among gram‐negative bacteria, whereas mice receiving SsoPox had the fewest associations. Conclusions Overall, our study establishes proof of concept that QQ is a targetable strategy for microbial control in vivo. Further characterization and dosage optimization of QQ enzymes are necessary to harness their therapeutic capability for the treatment of chronic microbial‐associated diseases.

Background Human adipose‐derived stem cells (hADSCs) are seed cells with application prospects in cartilage repair. However, the mechanism of hADSC chondrogenic differentiation is still unclear. This study identifies a novel circRNA, circNR3C2, which is significantly upregulated during the chondrogenic differentiation of hADSCs. Methods To analyze their role in hADSC chondrogenic differentiation, hADSCs were separated and identified by flow cytometry. Thereafter, we conducted Alcian Blue staining to assess chondrogenic differentiation levels. Additionally, RT‐qPCR was carried out to detect levels of the cartilage‐related genes COL2, Aggrecan and SOX9. Moreover, overlapping target SOX9 and circNR3C2 miRNAs were detected by bioinformatics and luciferase analyses. Finally, the role of circNR3C2 was confirmed in vivo using animal models. Results We confirmed that the cell surface receptors CD44, CD90 and CD105 were positively expressed on hADSCs, and their cartilage differentiation levels dramatically increased after 2 weeks. Expression of the cartilage‐related genes COL2 and Aggrecan and circNR3C2 also markedly increased. CircNR3C2 overexpression enhanced cartilage differentiation of hADSCs, while up‐regulating COL2, SOX9 and Aggrecan. Bioinformatics analysis identified hsa‐miR‐647 as the target miRNA of circNR3C2 and SOX9. Hsa‐miR‐647 overexpression in hADSCs can antagonize the effect of circNR3C2 on chondrogenic differentiation, and reverse its effect on regulating the expression of COL2, Aggrecan, and SOX9. We also showed that hADSCs overexpressing circNR3C2 promote cartilage repair in vivo. Conclusions We show that circNR3C2 modulates SOX9 expression to promote hsa‐miR‐647‐mediated hADSC chondrogenic differentiation; targeting circNR3C2 may help to develop new treatments to manage cartilage‐related disorders.

Background Alzheimer's disease (AD) and lung cancer are leading causes of mortality among the older population. Epidemiological evidence suggests an antagonistic relationship between them, whereby patients with AD exhibit a reduced risk of developing cancer and vice versa. However, the precise mechanism by which AD antagonizes lung cancer progression warrants further elucidation. Methods To this end, we established a co‐morbidity model using 5xFAD transgenic mice induced with the carcinogen urethane. We visualized and quantified surface lung tumor colonies, assessed pathological parameters associated with lung cancer and AD using histopathological analysis, and employed single‐cell sequencing and molecular pathological analyses to explore the mechanisms by which AD confers resistance to lung cancer. Results Our findings revealed a significant reduction in lung tumor incidence in the AD group compared with that in the wild‐type (WT) group. The results indicated a close association between AD‐induced inhibition of lung tumor progression and iron homeostasis imbalance and increased oxidative stress. Moreover, greater CD8⁺ T cytotoxic lymphocyte and effector natural killer cell infiltration in the lung tumor tissues of AD mice and enhanced CD8⁺ T cytotoxic lymphocyte‐mediated killing of target cells may be the primary factors contributing to the inhibition of lung tumor growth in the presence of AD. Conclusion This study identified essential mechanisms through which AD suppresses lung tumorigenesis, thereby providing targets for potential therapeutic interventions in these diseases.

Background This study investigated the impacts and mechanisms of yunweiling in the management of Functional Constipation (FC) using network pharmacology and experimental research. Methods Using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Genecard, and Online Mendelian Inheritance in Man (OMIM) databases, a potential gene target for yunweiling in treating FC was found. A pharmacological network was built and viewed in Cytoscape. A protein interaction map was created with STRING and Cytoscape. ‘clusterProfiler’ helped uncover its mechanism. Molecular docking was done with AutoDock Vina. In a constipation mouse model, Western blot was used to assess yunweiling's effectiveness. Results To investigate yunweiling's therapeutic effects on FC, we employed a loperamide‐induced constipation model. Successful model establishment was confirmed by first black stool time, reduced stool output, and impaired gastrointestinal motility. Yunweiling treatment, especially at high and medium doses, significantly alleviated constipation symptoms by reducing first black stool time, increasing stool output, and enhancing gastrointestinal motility. HE staining revealed yunweiling's ability to restore colon tissue structure. Yunweiling modulated the expression of key proteins TP53, P‐AKT, P‐PI3K, RET, and Rai, implicating its involvement in the PI3K‐Akt signaling pathway. Comparative analysis showed yunweiling to be more effective than its individual components (shionone, β‐sitosterol, and daucosterol) in improving constipation. The combination of yunweiling with TP53 and PI3K‐Akt inhibitors further enhanced its therapeutic effects, suggesting a synergistic mechanism. Conclusions The integration of network pharmacology and experimental investigations indicated the effectiveness of yunweiling in managing FC, offering essential theoretical support for clinical application.

Background The composition of the intestinal flora and the resulting metabolites affect patients' sleep after surgery. Methods We intended to elucidate the mechanisms by which disordered intestinal flora modulate the pathophysiology of postoperative sleep disturbances in hosts. In this study, we explored the impacts of anesthesia, surgery, and postoperative sleep duration on the fecal microbiota and metabolites of individuals classified postprocedurally as poor sleepers (PS) and good sleepers (GS), as diagnosed by the bispectral index. We also performed fecal microbiota transplantation in pseudo‐germ‐free (PGF) rats and applied Western blotting, immunohistochemistry, and gut permeability analyses to identify the potential mechanism of its effect. Results Research finding shows the PS group had significantly higher postoperative stool levels of the metabolites tryptophan and kynurenine than the GS group. PGF rats that received gut microbiota from PSs exhibited less rapid eye movement (REM) sleep than those that received GS microbiota (GS‐PGF: 11.4% ± 1.6%, PS‐PGF: 4.8% ± 2.0%, p < 0.001). Measurement of 5‐hydroxytryptophan (5‐HTP) levels in the stool, serum, and prefrontal cortex (PFC) indicated that altered 5‐HTP levels, including reduced levels in the PFC, caused sleep loss in PGF rats transplanted with PS gut flora. Through the brain–gut axis, the inactivity of tryptophan hydroxylase 1 (TPH1) and TPH2 in the colon and PFC, respectively, caused a loss of REM sleep in PGF rats and decreased the 5‐HTP level in the PFC. Conclusions These findings indicate that postoperative gut dysbiosis and defective 5‐HTP metabolism may cause postoperative sleep disturbances. Clinicians and sleep researchers may gain new insights from this study.

Background Pregnancy affects learning and memory in women. Thus, to investigate the effects of pregnancy, the authors examined the brain electrophysiology of pregnant mice. Methods Using the whole‐cell patch‐clamp technique on isolated brain slices, we detected and compared the electrophysiological changes in the hippocampal CA1 (HIP CA1) region, medial prefrontal cortex (mPFC), and basolateral amygdala (BLA) among 15 pregnant and 15 nonpregnant mice. Results In pregnant mice, there was a trend toward an increase in the frequency of miniature excitatory postsynaptic currents (mEPSCs) (p = 0.092) and a trend toward a decrease in the amplitude of miniature inhibitory postsynaptic currents (mIPSC) (p = 0.071) in the HIP CA1. In the BLA, both the amplitudes of mEPSCs and mIPSCs were significantly reduced (p = 0.004 and 0.042, respectively). In the mPFC, the amplitudes of mEPSCs and hyperpolarization‐activated currents (Ih), as well as the frequencies of mIPSCs, were higher compared to nonpregnant mice (p = 0.035, 0.009, and 0.038, respectively). Conclusions In pregnant mice, the electrophysiological change in neurons in the mPFC and BLA might contribute to the cognitive and emotional changes during pregnancy. A trend toward electrophysiological change in the HIP CA1 revealed that the mechanism of cognitive change during pregnancy might differ from that of other conditions.

Human herpesvirus, a specific group within the herpesvirus family, is responsible for a variety of human diseases. These viruses can infect humans and other vertebrates, primarily targeting the skin, mucous membranes, and neural tissues, thereby significantly impacting the health of both humans and animals. Animal models are crucial for studying virus pathogenesis, vaccine development, and drug testing. Despite several vaccine candidates being in preclinical and clinical stages, no vaccines are current available to prevent lifelong infections caused by these human herpesviruses, except for varicella‐zoster virus (VZV) vaccine. However, the strict host tropism of herpesviruses and other limitations mean that no single animal model can fully replicate all key features of human herpesvirus‐associated diseases. This makes it challenging to evaluate vaccines and antivirals against human herpesvirus comprehensively. Herein, we summarize the current animal models used to study the human herpesviruses including α‐herpesviruses (herpes simplex virus type 1(HSV‐1), HSV‐2, VZV), β‐herpesviruses (human cytomegalovirus (HCMV), γ‐herpesviruses (Epstein–Barr virus (EBV)) and Kaposi's sarcoma herpesvirus (KSHV)). By providing concise information and detailed analysis of the potential, limitations and applications of various models, such as non‐human primates, mice, rabbits, guinea pigs, and tree shrews, this summary aims to help researchers efficiently select the most appropriate animal model, offering practical guidance for studying human herpesvirus.

Background Qingyangshen (Cynanchum otophyllum C.K. Schneid) is a folk drug for treating depression and other mental disorders induced by social defeat stress. Neuroplasticity in the hippocampus is essential for the modulation of cognition and emotion, and its impairment may contribute to the development and progression of depression. Our previous studies have found that Qingyangshen glycosides (QYS) can improve depression‐like behavior in social failure mouse models, mainly through PGC‐1α/FNDC5/BDNF signaling pathways activation, but its effects and mechanisms on hippocampal neuroplasticity remain unknown. Methods Chronic social defeat stress (CSDS) was used to induce social defeat in mice. Morphological changes in the hippocampus were observed by H&E staining and Golgi staining. Immunofluorescence double staining was used to detect the expression of synaptophysin (SYN) and postsynaptic density protein‐95 (PSD‐95), while western blot was employed to evaluate PSD‐95, SYN, and doublecortin (DCX) proteins. The pathological processing of social defeat and the therapeutic effects of QYS on it was confirmed through behavioral assessment associated with morphologic observation. Results During the whole study, the sucrose preference indices and OFT activity time of CSDS mice were significantly decreased (p ≤ 0.05), and the tail suspension immobility time was significantly increased (p ≤ 0.05), suggesting that the mice had significant depressive symptoms. Treatment with QYS (25, 50, and 100 mg/kg) significantly alleviated depressive symptoms in CSDS mice, which was demonstrated by significantly (p ≤ 0.05 or p ≤ 0.01) reducing the duration of tail‐hanging immobility and increasing the tendency of sucrose preference indices and OFT activity time. QYS treatment also significantly increased the expression of DCX, PSD‐95, and SYN proteins, which play a crucial role in depression. Conclusions QYS alleviated these symptoms by enhancing hippocampal neuroplasticity through upregulating the expression of synapse‐associated proteins (SAPs). The therapeutic mechanism of QYS may involve modulating the neuroplasticity of hippocampus neurons by altering the expression of SAPs.

Acquired immune deficiency syndrome (AIDS) is the name used to describe several potentially life‐threatening infections and disorders that happen when HIV has severely compromised the immune system. The primary effect of HIV is to decrease host immunity, exposing the host to external pathogens. The development of pharmaceutical drugs that directly cure the infection is crucial because of the current wide‐ranging epidemic of HIV. Most therapeutic anti‐HIV drugs are nucleosides. However, their high toxicity and potential for drug resistance restrict their use. Many of the most effective clinical drugs used to inhibit HIV, the activation of latent HIV, and AIDS have been obtained from natural sources. This review focuses on potential natural medicinal products for treating and managing HIV and AIDS. Notwithstanding, further clinical research studies are needed to understand the subject and its dynamics.

Background The mechanisms underlying cardiac remodeling in aortic valvular (AoV) disease remain poorly understood, partially due to the insufficiency of appropriate preclinical animal models. Here, we present a novel murine model of aortic regurgitation (AR) generated by trans‐apical wire destruction of the AoV. Methods Directed by echocardiography, apical puncture of the left ventricle (LV) was performed in adult male C57BL/6 mice, and a metal guidewire was used to induce AoV destruction. Echocardiography, invasive LV hemodynamic and histological examination were conducted to assess the degree of AR, LV function and remodeling. Results AR mice exhibited rapid aortic regurgitation velocity (424 ± 15.22 mm/s) immediately following successful surgery. Four weeks post‐surgery, echocardiography revealed a 54.6% increase in LV diastolic diameter and a 55.1% decrease in LV ejection fraction in AR mice compared to sham mice. Pressure‐volume catheterization indicated that AR mice had significantly larger LV end‐diastolic volumes (66.2 ± 1.5 μL vs. 41.8 ± 3.4 μL), reduced LV contractility (lower dP/dtmax and Ees), and diminished LV compliance (smaller dP/dtmin and longer Tau) compared to sham mice. Histological examination demonstrated that AR mice had significantly larger cardiomyocyte area and more myocardial fibrosis in LV tissue, as well as a 107% and a 122% increase of heart weight/tibial length and lung weight/tibial length, respectively, relative to sham mice. Conclusions The trans‐apex wire‐induced destruction of the AoV establishes a novel and efficient murine model to develop AR, characterized by significant eccentric LV hypertrophy, heart failure, and pulmonary congestion.

The laryngeal muscle evoked potential (LMEP) is a neurophysiological outcome parameter that guarantees integrity of the nerve‐electrode interface during experiments with vagus nerve stimulation (VNS). This paper discusses a large series of minimally invasive LMEP recordings in 46 female Lewis rats, implanted with a custom‐made VNS electrode around the left cervical vagus nerve. After a 3‐week recovery, LMEPs were recorded twice in each animal, with swapping the anode and cathode positions of the VNS electrode (polarity inversion). A VNS‐induced LMEP was identified as the initial negative peak wave post‐stimulation artifact, consistently recorded in all sweeps at a given stimulation output current. Latency was defined as the time from stimulation onset to this negative peak, and stimulation threshold as the lowest current showing a clear and reproducible LMEP. An LMEP response was shown by 37/46 animals (80.4%), with stimulation intensity threshold of 0.37 ± 0.27 mA and latency of 2.39 ± 0.45 ms. Administering the cathodic pulse phase first at the caudal electrode contact resulted in the shortest LMEP latencies (MWU: p = 0.049. 2.36 ± 0.43 ms vs. 2.41 ± 0.47 ms). Minimally invasive LMEP recording provides a feasible and reliable means for checking electrode functioning and correct implantation.

Background Reliable animal models are crucial to drug development for focal segmental glomerulosclerosis (FSGS), a rare kidney disease. Variability in success rates in literature and significant ethical concerns with animal welfare necessitate further optimization of adriamycin (ADR)–induced FSGS model developed on BALB/c mice. Methods High‐performance liquid chromatography (HPLC) was used to assess ADR stability in water and upon light exposure. To identify the optimal ADR level, single intravenous ADR injections with dosing levels from 10 to 17 mg/kg body weight were administered to BALB/c mice to induce FSGS‐like pathology. Body weight and proteinuria of FSGS mice were monitored and analyzed for FSGS model–associated morbidity. Animals were euthanized for hematological and kidney histological assessments 8 weeks post induction. To identify the suitable experiment time frame of the ADR‐induced FSGS mouse model, a longitudinal study was performed, with an 11‐week continuous monitoring of the symptoms. Results ADR was found to be unstable in aqueous media and light sensitive. A dosing level of 10.5 mg/kg of ADR was optimal for consistent FSGS mouse model induction on BALB/c strain, characterized by minimal mortality and sustained FSGS‐like symptoms. Findings from the longitudinal study suggest that 6 weeks post ADR induction may represent the peak of FSGS pathology severity in this mouse model. This time frame may be used for FSGS drug development projects. Conclusion Based on the outcome from this study, we identified the optimal ADR dosing level and model testing duration. A standard operating procedure (SOP) for the ADR‐induced FSGS mouse model was established to facilitate FSGS basic research and drug development.

Background Donor nerve selection is a crucial factor in determining clinical outcomes of facial reanimation. Although dual innervation approaches using two neurotizers have shown promise, there is a lack of evidence‐based comparison in the literature. Furthermore, no animal model of dual reinnervation has yet been published. This study aimed to establish such a model and verify its technical and anatomical feasibility by performing dual‐innervated reanimation approaches in Wistar rats. Methods Fifteen Wistar rats were divided into four experimental groups and one control group. The sural nerve was exposed and used as a cross‐face nerve graft (CFNG), which was then anastomosed to the contralateral buccal branch of the facial nerve through a subcutaneous tunnel on the forehead. The CFNG, the masseteric nerve (MN), and the recipient nerve were coapted in one or two stages. The length and width of the utilized structures were measured under an operating microscope. Return of whisker motion was visually confirmed. Results Nine out of the eleven rats that underwent surgery survived the procedure. Whisker motion was observed in all experimental animals, indicating successful reinnervation. The mean duration of the surgical procedures did not differ significantly between the experimental groups, ensuring similar conditions for all groups. Conclusions Our experimental study confirmed that the proposed reanimation model in Wistar rats is anatomically and technically feasible, with a high success rate, and shows good prospects for future experiments.