Abstract: Objective To investigate whether there are sex differences in inflammatory bowel disease ( IBD) among the offspring of mice with IBD. Methods BALB/ c female mice were randomly divided into Na?ve and DSS groups.The mice in the Na?ve group drank autoclaved water freely, and the DSS group freely drank 2% dextran sodium sulfate(DSS) for 7 days before it was replaced with autoclaved water for 10 days. A total of 3 ~ 4 cycles were applied, and the IBD female mice were paired with healthy male mice in cages. When the pups were 8 weeks old, they were divided into the Con group and IBD group. The Con group drank autoclaved water freely for 7 days, and the IBD group drank 3% DSS for 7 days. During the modeling period, disease activity index was scored by monitoring body weight, fecal consistency, and the presence of blood in stool every day. Pathological sections were taken to observe changes in goblet cells and the mucus layer of colon tissues. The concentrations of interleukin ( IL)-6, IL-1β, IL-33, and IL-10 in the colon were detected by enzyme-linked immunosorbent assay. Real-time quantitative PCR was used to determine the mRNA expression levels o tight-junction proteins and MUC-2 in the colon. Results Compared with female IBD mice, male IBD mice had higher DAI scores, significantly shorter colons, larger amounts of inflammatory infiltrate, more crypt abnormalities, and a higher absence of goblet cells in the colon; their relative mRNA expression of occludin mRNA was significantly reduced, levels of IL-6 and IL-33 were significantly increased, and level of IL-10 was significantly decreased. Conclusions The symptoms of colitis in the offspring of IBD mice were more severe in male than in female mice, a result that was mainly attributed to the more severely impaired intestinal epithelial barrier function in males.

Abstract: Objective To investigate the effect of fear-induced stress during pregnancy on the expression of glucose transporters (GLUT) in the placenta, providing evidence for the theory of fetal damage caused by fear-induced stress during pregnancy. Methods Twenty pregnant Wistar rats were randomly divided into a control group and a model group of 10 rats each. In the model group, a fear-induced stress model was established using the modified bystander electroshock method for 20 days. After the experiment, the number of offspring and the weights of the placenta and fetal rats were measured, and the placental efficiency was calculated. Transmission electron microscopy was used to observe the morphological changes of placental cells. Bioinformatics analysis was performed to screen for differential genes in placentas affected by pregnancy stress-phobia, and gene set enrichment analysis was performed. Protein immunoblotting ( Western Blot), Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR), and immunohistochemistry were used to detect the expression levels of GLUT1, GLUT3, GLUT6, and GLUT7 proteins and genes. Results The placental efficiency was significantly reduced in the model group compared with that in the control group. The result of transmission electron microscopy in the model group showed that the placental microvilli were sparse and short and that the mitochondria and endoplasmic reticulum were swollen. Gene set enrichment analysis revealed that placental genes were significantly enriched in cellular glucose homeostasis in the model group compared with those in the control group. The result of Western Blot, Real-time PCR, and immunohistochemistry indicated a decrease in both the protein and gene expression levels of GLUT1, GLUT6, and GLUT7 in the placenta of pregnant rats. Conclusions Prenatal exposure to fear-induced stress may lead to adverse pregnancy outcomes. These adverse outcomes are potentially associated with reduced levels of three key GLUTs in the placenta: GLUT1, GLUT6, and GLUT7.

Abstract: Objective To clarify the changes in the microRNA (miRNA) expression profile of rat alveolar lavagefluid-derived exosomes after acute ozone exposure and analyze its potential relationship with lung injury. Methods Adult male Wistar rats were exposed to 0 ppm, 0. 5 ppm, and 2 ppm ozone for 6 h in a gas exposure chamber. After 24 h, the rats were anesthetized, the alveolar lavage fluid was collected, and exosomes were extracted. Differentially expressed miRNAs in exosomes were detected by miRNA-seq technology. miRDB databases were used to predict the target genes of the top 10 differentially expressed miRNAs among the groups, and Gene Ontology function enrichment and KEGG Pathway analyses were performed on the target genes. The differentially expressed miRNAs with high expression abundance and good uniformity were verified by quantitative reverse-transcription PCR ( qRT-PCR). Results Compared with the findings in the 0 ppm group, there were 8 up-regulated and 55 down-regulated miRNAs in the 0. 5 ppm group, and 32 up-regulated and 61 down-regulated miRNAs in the 2 ppm group. Compared with the findings in the 0. 5 ppm group, 90 exosomal miRNAs were up-regulated and 74 were down-regulated in the 2 ppm group. The target genes of top 10 differentially expressed miRNAs were mainly involved in regulating DNA binding, heterocyclic compound binding, and affecting transcription, and were closely related to cancer, MAPK, PIK3-AKT, TNF, ErbB, mitophagy, and FoxO signaling pathways. The qRT-PCR showed that the sequencing result were basically reliable. Conclusions Acute ozone exposure can lead to changes in the expression profile of exosomal miRNA in the alveolar lavage fluid of rats. Differentially expressed miRNAs may be involved in pathological processes, such as lung tissue inflammation and lung injury, caused by ozone exposure.

Abstract: Objective To investigate the inhibition of gigantol on corneal neovascularization (CNV) in rats after corneal alkali burn. Methods Animal models of corneal alkali burn were made with SD rats, which were divided into normal control group, model control group, low-concentration gigantol group, high-concentration gigantol group and aflibercept group, with 10 rats in each group. The rats in low-concentration gigantol group, high-concentration gigantol group and aflibercept group were treated with 2. 5 mg / 0. 05 mL gigantol, 5 mg / 0. 05 mL gigantol, and 2 mg / 0. 05 mL aflibercept by subconjunctival injection after modeling. The CNV, corneal opacity score, and thickness of the cornea were observed and compared on the 3rd, 7th, and 14th days after alkali burn. The ratio of CNV area to corneal area was calculated. On the 14th day, all rats were sacrificed. Hematoxylin and eosin staining and immunohistochemistry were used to detect the expression of CD34 and VEGF. The protein expression of VEGF, IL-1β, and TNF-α was detected by ELISA. Results On the 7th and 14th days after alkali burn, the percentages of CNV to total corneal area in low-concentration gigantol group, high-concentration gigantol group and aflibercept group were significantly smaller than those in model control group (all P<0. 05). On the 14th day, the corneal opacity score was lower in high-concentration gigantol group than model control group (P<0. 05). The corneal thickness in model control group and low-concentration gigantol group were significantly greater than that in normal control group ( all P<0. 001). However, the corneal thickness in highconcentration gigantol group and aflibercept group were not significantly different from that in normal control group (all P>0. 05). In addition, the protein expression of VEGF, IL-1β, and TNF-α in corneal tissues in low-concentration gigantol group, high-concentration gigantol group and aflibercept group were significantly lower than that in model control group (all P<0. 01). Conclusions Gigantol administration by subconjunctival injection can inhibit the formation of CNV in rats after alkali burn and promote absorption of the corneal edema.

Abstract: Objective To conduct basic research on the structure and function of the heart in cynomolgus monkeys older than 10 years to provide data for animal selection in elderly disease research. Methods A total of 144 cynomolgus monkeys > 10 years old were selected as research subjects, including 37 females and 66 males in the 10 ~ 15 years group, and 21 females and 20 males in the 16 ~ 20 years group. Basic data on cardiac structure and function in middle-aged and elderly cynomolgus monkeys were obtained through comparative analysis of general indicators ( body mass index, blood pressure, and heart rate), blood biochemical indicators ( blood glucose, blood lipids, and ion indicators), and cardiac structure and function indicators. Results General indicators for the 10 ~ 15 years and 16 ~ 20 years groups were compared. As age increased, the blood pressure and heart rate of female and male monkeys increased, and there was a significant difference in blood pressure changes between male monkeys. A comparison of two sets of blood biochemical indicators showed that, as age increased, blood glucose, triglycerides, total cholesterol, low-density lipoprotein cholesterol, blood calcium, blood sodium, and blood potassium increased, while lactate dehydrogenase decreased, in female and male monkeys. Among these, blood glucose, triglycerides (males), total cholesterol, high-density lipoprotein cholesterol (males), low-density lipoprotein cholesterol ( males), blood calcium, blood sodium, blood potassium, and lactate dehydrogenase showed significant changes. A comparison of cardiac contractile function between the two groups showed that, as age increased, the anterior and posterior diameters of the left atrium significantly decreased in both female and male monkeys. Female monkeys showed a significant decrease in the interventricular septal end systolic diameter, left ventricular end diastole and systolic diameters, left ventricular end diastolic and systolic volumes, and left ventricular mass index, while no significant changes were seen in male monkeys. A comparison of diastolic function between the two groups showed that, as age increased, the late diastolic velocity of the mitral valve decreased significantly in male monkeys, while the early diastolic velocity of the left ventricular sidewall increased significantly in female monkeys. Correlation analysis was conducted between the metabolic indicators and the cardiac structure and function indicators of female and male monkeys.The correlations between metabolic indicators and cardiac structure and function indicators were weak in female monkeys,for which the maximum absolute Γ value did not exceed 0. 39. However, the correlations between metabolic indicators and cardiac structure and function indicators were relatively strong in male monkeys, for which the maximum absolute Γ value reached 0. 66. Conclusions Based on ultrasound analysis combined with metabolic indicators, the heart function of cynomolgus monkeys was studied, and basic data related to the structure and function of the heart in middle-aged and elderly cynomolgus monkeys were obtained. As age increased, blood glucose and lipid indicators increased in cynomolgus monkeys, while cardiac systolic and diastolic functions show a downward trend, similar to changes in middle-aged and older adult human populations. These data provide support for animal selection in research on age-related diseases related to heart function.

Abstract: Objective Network pharmacology and animal experiments were performed to study and verify the therapeutic effect of Rhizoma Chuanxiong volatile oil on angina pectoris. Methods Volatile oil components were screened using steam distillation, gas chromatography-mass spectrometry, and oral bioavailability. Targets of these components were identified using Pubchem, SwissTarget, DisGeNET, and DrugBank databases as well as R language. Angina pectorisrelated targets and intersection targets were obtained. Protein-protein interactions were analyzed using the STRING database. The ClusterProfiler package in R was used to analyze the gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment of the intersecting targets, and Cytoscape was used to construct herbal-component-target-pathwa networks. Molecular docking analysis was conducted using AutoDock Vina 1. 2. 3, Pymol 3. 0, and Discovery Studio 2016 software to evaluate the affinity between key targets and the main volatile oil components. Finally, the therapeutic effect of Rhizoma Chuanxiong volatile oil on angina pectoris was verified by animal experiments. Results In total, 10 volatile oil components and 22 key targets were identified. They were closely related to neurotransmitters, receptors on synaptic membranes, material metabolism, neuroactive ligand-receptor interaction, retinol metabolism, and drug metabolismcytochrome P450 pathways. The molecular docking result showed that 3-butylidenephthalide, alpha-selinene, transligustilide, and other volatile oil components combined with several key targets play therapeutic roles. Animal experiments showed that the volatile oil of Rhizoma Chuanxiong can regulate the ejection fraction, fractional shortening, stroke volume,and left ventricular internal diameter in systole and the activities of lactate dehydrogenase, creatine kinase and aspartate aminotransferase; promote the expression of ADRA1A and CHRM5 proteins in damaged cardiomyocytes; improve the state of myocardial fibers; reduce intercellular space; and reduce inflammatory cell infiltration. Conclusions The volatile oil of Ligusticum wallichii can effectively protect damaged myocardial tissue and thus has a role in treating angina pectoris.

Abstract: Objective To explore the preventive and therapeutic effects of Caryopteris incana decoction in a rat model of primary dysmenorrhea with cold coagulation and blood stasis syndrome. Methods Sixty healthy specific pathogen-free SD female rats were randomly divided into six groups of ten rats each: normal group, model group, ibuprofen group, C. incana high-dose group, C. incana medium-dose group, and C. incana low-dose group. All groups except the normal group were treated with cold stimulation combined with estradiol benzoate and oxytocin to establish a rat model of primary dysmenorrhea with cold coagulation and blood stasis syndrome. On the fifth day of modeling, the rats were intragastrically administered the study drugs for 10 days. Their symptoms were observed and recorded. The writhing response and hemorheological indices were measured. The serum levels of TXB₂ , 6-keto-prostaglandin F_1α ( 6-KetoPGF_1α ), estradiol ( E₂ ), and progesterone ( PROG) were measured. Additionally, the levels of prostaglandin F_2α (PGF_2α ), prostaglandin E₂(PGE₂ ), nitric oxide (NO), and calcium (Ca²⁺) in the uterine tissues were measured. The organ indices of the uterus and ovary were calculated, and histopathological changes were observed. Results Compared with the normal group, the rats in the model group showed obvious symptoms of cold coagulation and blood stasis syndrom and writhing reaction. The morphology of uterus and ovary showed obvious hyperplasia, inflammation, edema and other lesions. The plasma viscosity, packed cell volume and whole blood viscosity were significantly increased (P<0. 01). The serum levels of thromboxane B₂ and E₂and the E₂ / PROG ratio were significantly increased (P<0. 01), the levels of 6-Keto-PGF_1α and PROG were significantly decreased (P<0. 01). The uterine index and ovarian index were significantly increased (P<0. 01). The levels of PGF2α and Ca ²⁺ and the PGF_2α / PGE₂ ratio in uterine tissue were significantly increased (P<0. 01), while the levels of PGE₂ and NO were significantly decreased (P<0. 01). Compared with the model group, Caryopteris incana significantly improved the symptoms of model rats, improved the morphological lesions of the uterus and ovary, prolonged the latency time of the writhing reaction, and reduced the number of writhing episodes (P<0.01);significantly reduced the plasma viscosity, packed cell volume, and whole blood viscosity ( P<0.01); significantly reduced the serum levels of TXB₂ and E₂ and the E₂ / PROG ratio, increased the serum levels of 6-Keto-PGF_1α and PROG, and reduced the uterine and ovarian indices (P<0. 01,P<0. 05); significantly reduced the levels of PGF_2α and Ca ²⁺and the PGF_2α / PGE₂ ratio in uterine tissue (P<0. 01, P<0. 05); and significantly increased the levels of PGE₂ and NO in the uterine tissue (P<0. 01). Conclusions Caryopteris incana decoction can effectively improve the clinical symptoms of primary dysmenorrhea in rats with cold coagulation and blood stasis syndrome, and it has a good control effect. Its mechanism may be correlated with the levels of TXB₂, 6-Keto-PGF_1α , E₂ , and PROG in serum and PGF_2α , PGE₂ , NO, and Ca²⁺ in uterine tissue.

Abstract: Objective To observe the neuropsychiatric behavioral performance of kainic acid ( KA)-induced epilepsy rats; investigate gender differences in acute seizure and behavioral performance tasks relating to sense, motor,learning, and memory in the remission phase; and explore the potential neurobiological mechanisms of action. Methods Healthy SD rats aged 4 weeks were randomly divided into control and model groups, with 22 rats in each group (11 males and 11 females). An epileptic rat model was induced by intraperitoneal injection of KA. Seizure latency and frequency within 2 hours of KA injection were observed, seizure grade was assessed using the Racine grade standard, and a cortical electroencephalogram (EEG) was recorded. Behavioral performance was observed in a series of tasks including open field testing, balance beam walking, elevated plus maze, Y-maze, and novel object recognition. The level of GABA in the hippocampus was detected by ELISA, injury to hippocampal neurons was observed by Nissl staining, and the protein expression of synapsin-1 and synaptotagmin 1 in the hippocampus were detected by Western Blot. Results Both male and female rats presented typical epileptic behaviors after KA injection. However, compared with the effects in males, the latency of the first seizure (p=0. 014) and IV ~ V grading in female model rats were more pronounced (P<0. 01), and the frequency of epileptic seizures within 2 hours was significantly reduced (p=0. 019). In the open field testing, KAinduced epileptic rats presented more motor but fewer hedonic behaviors, as indicated by the decrease in total movement distance in the central area, compared with the control group. Moreover, grooming frequency was significantly reduced in the female model rats compared with not only that in the control but also that in male model rats (P<0. 01). The model rats spent more time completing and had a higher score in the balance beam walking task, indicating their poorer stability and balance. In the elevated plus maze, the exploration times of male model rats in the closed arm was increased. The preference index of rats for the novel arm or object decreased in the Y-maze and novel object recognition, suggesting impairments to their learning and memory abilities. Moreover, neuronal injuries were found in the hippocampus of the model rats that were accompanied with a decline in GABA concentration and protein expression of synapsin-1 and synaptotagmin 1, with no gender differences. Conclusions Intraperitoneal injection of KA successfully induced an epilepsy rat model. However, there was a gender difference in the characters of acute seizures and performance of sensory, motor, and learning memory during epileptic remission. There was no gender differences in the hippocampal GABA concentration or expression of synaptic plasticity-related proteins, and thus no evidence was found for the mechanisms underlying the gender differences.

Abstract: Objective In this study, we aimed to use network pharmacology techniques to predict the key targets of a prescription of Ziziphi spinosae semen formula (ZSSF) compound for depression, and to verify its mechanism of action using a zebrafish model of rifampicin-induced depression. Methods The drug targets of ZSSF were retrieved from the TCMSP database, and the target names were corrected using the UniProt database. Depression-related targets were identified using the GeneCards, OMIM, and NCBI databases. Protein-protein interaction information for the shared targets was predicted using the STRING database. The collected data were then analyzed using the Metascape database to determine GO and KEGG pathway enrichment, and the result were visualized using microbiotics. Behavioral experiments and reverse-transcription quantitative PCR experiments were conducted to verify the therapeutic effects of ZSSF on a zebrafish depression model induced by risperdal. Results 188 targets were screened to find the interactions between depression and ZSSF. The protein-protein interaction result showed that ZSSF primarily targeted TNF-α, IL-2, IL-6, IL1β, and IL-10 to produce its antidepressant effect. KEGG pathway enrichment analysis revealed that ZSSF exerted its effects on depression through various signaling pathways, including the TNF, PI3K-Akt, and cGMP-PKG signaling pathways. The result of the animal experiments showed that the treatment groups given high, medium, and low doses of ZSSF exhibited significant improvements in movement distance under acoustic and light stimulation compared with the model group(P<0. 05). The speed of movement of the treatment groups was also significantly faster(P<0. 01). Additionally,the mRNA expression levels of TNF-α, IL-2, IL-6, IL-1β, and IL-10 were up-regulated in the brain tissues of zebrafish in the high-, medium-, and low-dosage groups of ZSSF compared those in the model group ( P<0. 001). Conclusions ZSSF exerts its antidepressant effect through multiple components and targets, and its antidepressant effects may be associated with its inhibition of inflammatory factors.

Abstract: Objective To review the characteristics of animal models of sequelae of pelvic inflammatory disease (SPID) to provide a reference for standardizing the modeling process and improving the modeling rate. Methods Literature relevant to animal models of SPID from the past 40 years was searched, and animal species, modeling method, modeling cycles, modeling substances, positive control drugs, and evaluation indexes were summarized and analyzed. Results A total of 243 study manuscripts were included, most of which induced the SPID model in rats via the phenol paste or microbial infection method. The modeling cycles were typically between 14 and 16 days, and the success of the models was mostly determined by pelvic tissue morphology observation and pathological HE staining. Few research reports have focused on the traditional Chinese medicine(TCM) disease combination model. Conclusions No consistent criteria have been established for SPID animal model modeling, and thus it is recommended that researchers evaluate changes to animal behavior, pelvic histomorphology, and pathology. TCM syndrome modeling lacks effective method and evaluation standards, which need further research and development. Finally, the selection and use of positive-control drugs need to be further explored and perfected.

Abstract:The annual incidence of neurodegenerative disease has been increasing with the aging of the global population, seriously affecting the quality of life of elderly patients and imposing a heavy burden on society. Glycyrrhetinic acid, which inhibits neuroinflammation and protects neurons, is one of the main active ingredients of the traditional Chinese medicine Glycyrrhiza glabra. Increasing numbers of studies are focusing on the mechanism of action of glycyrrhetinic acid and its derivatives in neurodegenerative disease. This review summarizes studies on the effects and mechanisms of action of glycyrrhetinic acid and its derivatives in Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, multiple sclerosis, and cerebellar atrophy. Additionally, the future applications of glycyrrhetinic acid and its derivatives in neurodegenerative disorders are discussed.

Abstract:Necroptosis is a regulated process of programmed cell death independent of aspartic acid-specific cysteine protease, which can induce inflammation. Studies have shown that necroptosis is closely related to the progression and prognosis of pancreatic disease and plays an important two-way regulatory role in its progression. Related necroptosis inhibitors and inducers are expected to be used in the treatment of pancreatic disease. We herein review the mechanism of necroptosis and its role in the progression of pancreatic disease to provide a new understanding of the pathogenesis and treatment of pancreatic diseases and offer a theoretical basis for the research and development of targeted drugs.

Abstract:Parkinson’s disease is the second most common neurodegenerative disease in middle-aged and elderly people. It is characterized by a long disease course and complex treatment process, introducing great challenges to society.Behavioral changes in animal models of Parkinson’ s disease can intuitively reflect the modeling situation of experimental animals and the effects of drug interventions. Therefore, selecting standardized animal models and appropriate behavioral assays is fundamental for both understanding the mechanisms of Parkinson’s disease and developing anti-Parkinson drugs.In this paper, we summarize the method of behavioral experiments of Parkinson’s disease using mice and rats at home and abroad and systematically summarize the experimental equipment, experimental method, evaluation indexes, and precautions of commonly used Parkinson’s behavioral experiments. We also provide an overview of the commonly used animal models of Parkinson’s disease and analyze their modeling mechanisms, alignment with the clinical features of Parkinson’s disease, and respective advantages and disadvantages. This analysis will help researchers in choosing appropriate animal models of Parkinson’s disease and behavioral testing method according to the purpose of the study.