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Portal vein thrombosis （PVT） refers to thromboembolism that occurs in the extrahepatic main portal vein and/or intrahepatic portal vein branches. PVT is the result of the combined effect of multiple factors， but its pathogenesis remains unclear. Animal models are an important method for exploring the pathophysiological mechanism of PVT. Based on the different species of animals， this article reviews the existing animal models of PVT in terms of modeling methods， principles， advantages and disadvantages， and application.
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Craniocerebral injury with seawater immersion is a special kind of compound injury, with low temperature, high permeability, high alkali, high salt content, and bacterial infection being the main causes. The injury is also characterized with complex damage mechanisms, difficulty to treat, and poor prognosis. At present, the damage mechanisms of craniocerebral injury with seawater immersion are mainly studied by establishing the experimental animal models at the levels of tissue, cell, organelle, molecule, etc. However, the craniocerebral injury with seawater immersion is more complex than the simple onshore craniocerebral injury, therefore, a stable disease model is not easy to construct. Most researches on the specific injury mechanisms are relatively single and one-sided, with many different views in existence, and the damage mechanisms of craniocerebral injury with seawater immersion have hitherto not been clear. The authors reviewed the research progress in the damage mechanisms of craniocerebral injury with seawater immersion, in order to promote the in-depth study of the mechanism of craniocerebral injury with seawater immersion and provide reference for its clinical treatment.
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Objective:To examine the impact of berberine on polycystic ovary syndrome (PCOS) in mice, and to investigate the effects of berberine on the intestinal flora and the intestinal flora on PCOS.Methods:A mouse model of PCOS was established by administering dehydroepiandrosterone in combination with high fat diet, and the mouse model was given a berberine treatment. The study consisted of a blank control group (C group), a PCOS model group (M group) and a berberine treatment group (T group). During the experiment, the mice were closely monitored through timed body weight measurements and estrous cycle monitoring; intraperitoneal glucose tolerance test and insulin tolerance test were done. Upon completion of the pharmacological intervention, the wet weights of liver, ovary and fat deposits of mice were assessed and subjected to HE staining to confirm the success of PCOS modeling and the efficacy of berberine. Additionally, fecal samples were analyzed for intestinal flora through 16S rRNA analysis.Results:The PCOS model was established successfully, berberine alleviated the disturbance of estrous cycle in mice, and significantly alleviated fat accumulation and metabolic abnormalities of glucose in mice. The cross-sectional area of fat pad cells in T group was (2 858±146) μm2, which was significantly lower than that in M group [(9 518±347) μm2], and the difference was statistically significant ( P<0.001). The blood glucose levels in T group were significantly lower than those in M group ( P<0.05). The composition and structure of intestinal flora in mice of M group with PCOS (compared with C group) and in mice of T group after berberine intervention (compared with M group) were significantly altered. However, alpha diversity did not change significantly among three groups ( P>0.05). Conclusion:Berberine could alleviate PCOS by intervening in the alterations of gut microbiota.
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Objective To determine the improved effect of methylene blue(MB)on cognitive func-tion in brain-inflammatory-aging rats and investigate the underlying mechanism.Methods A total of 38 healthy 12-month-old SD rats were randomly divided into healthy control group,lipopo-lysaccharide(LPS)group,MB vehicle group and MB group,with 8 rats in the control and 10 rats in the other three groups.LPS was injected into the fourth ventricle with aid of a subcutaneous sustained release pump to establish a rat model of brain chronic inflammatory aging.MB of 0.5 mg/(kg·d)was added into the pump in the rats from the MB group.T-maze test and new object recognition test were employed to evaluate the learning and memory abilities of the rats.The acti-vation of microglia and astrocytes in the hippocampal CA1 region of the rats was detected by im-munofluorescence assay.The release of inflammatory factors IL-1β and IL-6 was measured by ELISA,and neuronal death in the CA1 region was assessed by neuronal nuclei(NeuN)fluores-cence staining.Results There was no significant difference in the exploration time for new and old objects between the LPS group and the MB solvent group(P＞0.05).The MB group spent significantly longer time in exploring the new objects than the old object(22.50±4.32 s vs 11.60± 3.01 s,P=0.000).The alternating selection rate of new arm and expression level of NeuN antigen were significantly decreased,and the expression levels of ionized calcium binding adaptor mole-cule-1(Iba-1)and glial fibrillary acidic protein(GFAP)and the contents of IL-1β and IL-6 were obviously increased(P＜0.05)in the LPS group and the MB vehicle group than the healthy con-trol group.Compared with the MB vehicle group,the MB group had notably increased alternating selection rate of new arms and higher NeuN expression level,and decreased Iba-1 and GFAP ex-pression and IL-1β and IL-6 contents(P＜0.05).Conclusion Subcutaneous administration of MB could significantly inhibit the damages of spatial learning and memory abilities in the LPS-induced brain chronic inflammatory aging rats.The mechanism may be closely associated with MB inhibi-ting inflammatory glial cells and protecting hippocampal pyramidal neurons.
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Objective:To observe and analyze depression-like behavioral performances of mouse models of vitiligo.Methods:Fifteen female C57BL/6 mice aged about 9 weeks were modeled for vitiligo. Whether the mouse models of vitiligo were successfully constructed or not was determined by macroscopy and full-thickness epidermal immunofluorescence staining of mouse tail tissues on day 23 after the start of the experiment; on day 8 (pre-modeling stage) and day 21 (early modeling stage), the elevated plus maze test and the open field test were used to evaluate the behavioral performances of the mice, including the number of entry into the open arms, percentages of time spent in the open arms, percentages of time spent in the central area and total distance traveled, aiming to assess whether depression-like behaviors were exhibited in the mouse models of vitiligo. To further clarify the degree of the impact of vitiligo modeling on the depression-like state in mice, 20 female C57BL/6 mice were equally divided into 2 groups: vitiligo modeling group and vitiligo modeling + chronic restraint stress group; the mice in the vitiligo modeling + chronic restraint stress group were subjected to chronic restraint stress on day 9, that is, these mice were placed in centrifuge tubes and restrained for about 6 hours every day for 28 consecutive days; on days 7, 22, 29 and 38 after the start of vitiligo modeling, the above-mentioned behavioral indicators were determined by the elevated plus maze test and open field test in the 2 groups. Repeated measurement data in a single group were compared before and after treatment by using paired t-test, and repeated measurement data at multiple time points were compared by using two-way repeated measures analysis of variance. Results:By macroscopy, the mice gradually developed well-defined white patches on the tail skin during vitiligo modeling, which were similar to the clinical manifestations of vitiligo patients; on day 23, full-thickness epidermal immunofluorescence staining of the mouse tail tissues was conducted and showed obvious infiltration of CD8 + T cells and a decrease in the number of Melan-A-positive epidermal melanocytes under a laser confocal microscope, which were consistent with typical pathological characteristics of vitiligo; based on the macroscopic results and immunofluorescence findings, a total of 12 mouse models of vitiligo were successfully constructed on day 23. The elevated plus maze test showed that the number of entry into the open arms and the percentages of time spent in the open arms were significantly lower in the 12 mouse models of vitiligo on day 21 (2.33 ± 1.78 times, 5.01% ± 5.27%, respectively) than in those on day 8 (10.75 ± 2.30 times, 29.20% ± 12.48%, t = 9.63, 6.36, respectively, both P < 0.001) ; the open field test showed that the percentages of time spent in the central area and total distance traveled were also significantly lower in the mouse models on day 21 (2.31% ± 1.53%, 2 518.31 ± 528.38 cm, respectively) than in those on day 8 (4.47% ± 2.65%, 3 533.45 ± 465.47 cm, t = 2.40, 5.47, P = 0.036, < 0.001, respectively). In the chronic restraint stress test, a total of 14 mouse models of vitiligo were successfully constructed on day 23, including 5 in the vitiligo modeling group and 9 in the vitiligo modeling + chronic restraint stress group. There were no significant differences in the number of entry into the open arms, percentages of time spent in the open arms, percentages of time spent in the central area, and total distance traveled between the vitiligo modeling group and the vitiligo modeling + chronic restraint stress group on days 7, 22, 29, and 38 ( F = 0.21, 0.20, 0.46, 2.35, P = 0.889, 0.893, 0.719, 0.134, respectively) ; moreover, all the above indicators significantly changed over time (all P < 0.001), except for the total distance traveled ( P = 0.422) . Conclusion:The mouse models of vitiligo developed depression-like behavior at the early modeling stage, and the degree of depression could not be further deepened by chronic restraint stress on the basis of vitiligo modeling.
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ObjectiveTo establish a nude mouse model of type 2 diabetes mellitus （T2DM） and pancreatic cancer that allows dynamic observation of tumor formation process and facilitates in vivo research. MethodsAt first， human pancreatic cancer PANC-1 cells were transfected with lentiviral vector GV260 to construct the pancreatic cancer cell line PANC-1-Luc with stable expression of firefly luciferase. Then， 36 specific pathogen-free nude mice were randomly divided into control group with 12 mice and model group with 24 mice （nude mice with T2DM and pancreatic cancer）. The mice in the control group were fed with breeding diet and were then given ectopic subcutaneous implantation of PANC-1-Luc cells， and those in the model group were first given high-fat diet and intraperitoneal injection of 1% STZ， followed by ectopic subcutaneous implantation of PANC-1-Luc cells. The fluorescence in vivo imaging system and the manual measurement method were used for simultaneous and dynamic monitoring of the growth of pancreatic cancer in nude mice in the two groups， and the tumor growth curve was plotted to investigate the correlation between fluorescence value and tumor volume. Subcutaneous tumors and pancreatic islets were observed under a microscope to verify whether the model was successfully established， and immunohistochemistry was used to measure the expression of Ki-67 in tumor tissue to investigate the influence of hyperglycemia on the growth of pancreatic cancer in nude mice. The independent-samples t test was used for comparison of normally distributed continuous data between groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups. ResultsThe optimal virus titer was determined as 5×107 TU/mL for the stable transfection of lentiviral vector in PANC-1 cells， and the optimal concentration selected with puromycin was 20 μg/mL， with an optimal selection time of 9 days. The fluorescence value of PANC-1-Luc cells was linearly and positively correlated with the number of cells， with the linear equation of y=42.56x－42 504 （r=0.977， P=0.004）. The blood glucose value of T2DM nude mice was 23.05 （19.25 — 26.40） mmol/L， with a blood glucose level of >11.1 mmol/L in each nude mouse， and there was a significant difference in blood glucose value between the T2DM nude mice and the control nude ［6.15 （5.20 — 7.30） mmol/L］ （Z=-8.45， P<0.001）. Compared with the control group， the model group had reductions in the number and volume of pancreatic islets， with irregular shapes and unclear boundaries， and pathological examination confirmed that the xenograft tumor was pancreatic cancer tissue， which showed that the model was established successfully. In the model group， there was a linear positive correlation between subcutaneous tumor size and fluorescence values， with the linear equation of y=232 348 691x－8 258 608 （r=0.911， P=0.031）. The model group had a significantly higher positive rate of Ki-67 than the control group （50.333%± 7.808% vs 15.917%±4.055%， t=13.55， P<0.001）， suggesting rapid tumor proliferation in the model group. ConclusionThe T2DM nude mouse model of pancreatic cancer established in this study can simulate the pathological process of the development and progression of pancreatic cancer in the context of T2DM and dynamically observe the influence of hyperglycemia on the growth of pancreatic cancer cells in vivo， thereby providing a new experimental vector for the in vivo study of the development and progression of pancreatic cancer in the context of T2DM.
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Cholestatic liver diseases (CLD) are a series of diseases due to impaired bile flow and accumulation of bile acid in the liver and/or systemic circulation caused by immune, genetic, and environmental factors. The pathogenesis of CLD remains unclear and CLD is difficult to treat. As a substitute for human diseases, animal models can provide a platform for exploring the etiology and pathogenesis of the disease and finding appropriate therapeutic targets. This article reviews the current research advances in the animal models of CLD.
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Objective:To investigate the effect of galectin-3 (gal-3) on microglia polarization after subarachnoid hemorrhage (SAH).Methods:C57BL/6 male adult mice were used to induce SAH or sham operation models. Gal-3 siRNA or negative control siRNA was injected into the lateral ventricle 48 h before the model was induced. After 24 h of model preparation, the SAH score, neurological function score, brain water content, and Evans blue exudate were measured. Western blot analysis was used to detect the expressions of M1 phenotypic markers (inducible nitric oxide synthase [iNOS], CD11b, tumor necrosis factor [TNF]-α) and M2 phenotype markers (CD206, YM1/2, arginase-1 [Arg1]).Results:After using Gal-3 siRNA to inhibit Gal-3, the neurological function score significantly increased, while the SAH score, brain water content, and Evans blue exudate significantly decreased ( P<0.001). Western blot analysis showed that the expressions of M1 phenotypic markers (iNOS, CD11b and TNF-α) in microglia were significantly decreased after Gal-3 inhibition, while the expressions of M2 phenotypic markers (CD206, YM1/2 and Arg1) were significantly increased ( P<0.001). Conclusion:Inhibition of Gal-3 expression can alleviate the early brain injury after SAH, and its mechanism may be associated with regulating the polarization of microglia from M1 to M2 phenotype.
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Due to the failure of multiple translational researches, Stroke Therapy Academic Industry Roundtable (STAIR) recommends the use of large animal models of focal cerebral ischemia for preclinical researches. Especially, stroke treatment has currently entered a new era of vascular recanalization. Large animals commonly used in acute ischemic stroke models include dogs, swine, sheep, and non-human primates, which can be used to simulate various aspects of cerebral ischemia and reperfusion (vascular recanalization) in patients. Although large animals have significant advantages due to their proximity to humans in anatomy and physiology, there are also issues with anatomical and physiological specificity and ethical limitations. This article summarizes the large animal ischemic stroke models prepared by craniotomy and endovascular intervention, hoping to help researchers select the most appropriate large animal ischemic stroke model, and then promote the development of stroke translational research.
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Vertebrobasilar dolichoectasia (VBD) is a cerebrovascular variant disease. Researches have shown that further development of VBD may lead to severe disability and even death. The pathogenesis of VBD is still unclear, and there is no specific clinical prevention and treatment scheme. Therefore, establishing a stable and reliable animal model helps to further understand the pathophysiological mechanisms and potential therapeutic targets of VBD. This article reviews the establishment methods and research progress of the available VBD animal models.
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Cerebral cavernous malformations (CCMs) are the common cerebrovascular malformation. Its incidence was 0.16%-0.5%. CCM can exist in both sporadic and familial forms, with the latter being inherited in an autosomal dominant manner. Its pathogenesis is associated with mutations in the CCM1, CCM2, and CCM3 genes. The somatic mutations of these genes are the basis for the occurrence of brain lesions. In order to explore the pathogenesis of CCM and identify therapeutic targets, various CCM animal models have been developed, providing assistance for the study of the pathological and physiological mechanisms of CCM. However, each CCM model has its own advantages, disadvantages, and applicability. Mice are the most commonly used animals to model CCM. Therefore, this article summarizes the characteristics and research progress of current murine CCM models.
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Establishing appropriate animal models of diabetes is of great importance for gaining in-depth understanding of the pathogenesis of diabetes, conducting diagnosis, and developing novel therapeutic interventions. Exploring methods for treating limb ischemia and comprehending its clinical pathophysiological progression is a significant approach achieved through the use of animal models of lower limb ischemia. In recent years, with the rapid advancements of various techniques, the establishment of lower limb arterial ischemia models and diabetes models has made considerable progress. The establishment of these models carries vital implications for researching and comprehending the mechanisms underlying lower limb arterial ischemia and the impact of diabetes. Successfully and reliably preparing these two models is crucial for elucidating the pathogenesis, pathophysiology, and treatment of lower limb vascular lesions associated with diabetes. This paper provides an overview of the research on the establishment methods of a diabetic rat model of lower limb ischemia, covering the principles, methods, and relevant parameters of establishing a diabetic rat model and a lower limb ischemia model in diabetes.
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Objective:To investigate the effects of modified Buzhong Yiqi Decoction on intestinal microflora in a rat model of chronic obstructive pulmonary disease. Methods:From April to June 2021, 60 specific pathogen-free Wistar rats were selected for this study. They were randomly divided into blank control, model, traditional Chinese medicine, and western medicine groups with 15 rats per group. Rat models of chronic obstructive pulmonary disease with lung and spleen deficiency were established in all groups except the blank control group. Rat models in the traditional Chinese medicine and western medicine groups were administered modified Buzhong Yiqi Decoction and synbiotics. Rat models in the model and blank control groups were identically administered 0.9% sodium chloride injection. After 7 days, the feces of rats in each group were collected for 16S rRNA sequencing of intestinal flora. Effective sequences were clustered to obtain operational taxonomic units for principal coordinate analysis, species composition analysis, and alpha diversity analysis. The effects of modified Buzhong Yiqi Decoction on the structure, diversity, and abundance changes of intestinal flora were analyzed. Results:The dominant bacteria in the traditional Chinese medicine and western medicine groups were Firmicutes, while the dominant bacteria in the blank control and model groups were Bacteroides. The dominant bacterial groups in each group were mainly Lactobacillus and Bacteroides. Alpha diversity analysis showed that the Shannon index in the community diversity indices of traditional Chinese medicine, western medicine, and blank control groups was (3.65 ± 0.35), (3.65 ± 0.36), and (3.59 ± 0.20), respectively, which were significantly higher than (3.37 ± 0.26) in the model group ( t = 2.49, 2.44, 2.60, all P < 0.05). There was no significant difference in the Shannon index among traditional Chinese medicine, western medicine, and blank control groups (all P > 0.05). The Sobs index of the traditional Chinese medicine, western medicine, and blank control group was (458.67 ± 73.11), (454.80 ± 95.13), and (525.93 ± 101.88), respectively, which were significantly higher than (337.40 ± 37.49) in the model group ( t = 5.72, 4.45, 6.73, all P < 0.05). The Sobs index in the blank control group was higher than that in the western medicine group. There was no significant difference in the Sobs index between blank control and traditional Chinese medicine groups and between western medicine and traditional Chinese medicine groups (both P > 0.05). Principal coordinate analysis revealed that compared with the blank control group, Actinomycetes decreased and Proteobacteria and Desulfurization bacteria increased at the phylum level in the model group, while compared with the blank control group, Bacteroides, Rombutzia,Trichospirillus, and Parabacteroides increased, and Prevotella, Clostridium, Brucella, and Ruminococcus decreased at the genus level. Compared with the western medicine group, Bacillus, Prevotella, Brucella, and Prevotellidae in the traditional Chinese medicine group increased, while Clostridium, Pectinobacter, Christensen, and Trichospirillus decreased in the traditional Chinese medicine group. There was a statistically significant difference in the composition of the bacterial population between groups (all P < 0.05). Conclusion:There is an imbalance in intestinal microecology in a rat model of chronic obstructive pulmonary disease. Modified Buzhong Yiqi Decoction can regulate the intestinal microecology environment, improve the structure of intestinal flora, and increase the diversity and abundance of intestinal flora.
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The pathogeneses of oral squamous cell carcinoma and most oral mucosal diseases are unclear. Therefore, establishing animal models with similar pathogeneses is significant for clinical prevention, diagnosis, and treatment of related diseases. At present, scholars have established animal models for different focuses. This paper aims to introduce the methods for establishing animal models of oral squamous cell carcinoma and common oral mucosal diseases, compare their advantages and disadvantages, and provide evidence for related basic research.
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Objective:To explore the effects of prenatal dexamethasone (DEX), postnatal pulmonary surfactant (PS) and respiratory support on the lung fluid clearance in premature rabbits at gestational age (GA) of 25-28 d (full term: 31 d) and their relationship with dynamic compliance of respiratory system (Cdyn), pulmonary morphology and other parameters.Methods:In our previous publications, premature rabbits were divided into four groups according to the intervention strategy: control group, PS-only group, DEX-only group and DEX+PS group in which data of several parameters including wet-to-dry lung weight ratio (W/D), Cdyn and volume density of alveoli (Vv) were retrieved and the lung tissue sections were scanned to recalculate the ratio of perivascular sheath to vascular sectional area (S/V) and lung injury scores-edema (LIS-E). W/D, LIS-E, S/V and Vv were adjusted for birth weight (BW) (divided by BW, represented as W/D/BW, LIS-E/BW, S/V/BW and Vv/BW) and mean Cdyn (Cdyn-m) was adopted. Based on the grouping of previous studies, the intervention groups in this study were divided as DEX group and non-DEX group, and PS group and non-PS group to analyze the influence of DEX and PS on the above parameters. Two independent samples t-test, one-way analysis of variance, LSD test, Kruskal-Wallis H test, Mann-Whitney U test and Pearson correlation analysis were used for statistical analysis. Results:A total of 196 newborn rabbits receiving mechanical ventilation after birth were included in this study. (1) Effects of DEX: compared with the non-DEX group, the DEX group showed increased W/D/BW (489±69 vs 421±113, t=－2.09), LIS-E/BW (188±57 vs 138±55, t=－2.61) and Vv/BW (20.1±4.9 vs 14.2±4.7, t=－3.60), but decreased S/V (0.33±0.23 vs 0.51±0.25, t=2.23) and S/V/W/D (0.05±0.03 vs 0.07±0.04, t=2.22) at 25 d of gestation; at 26 d of gestation, W/D/BW (472±76 vs 303±44, t=－8.75), LIS-E/BW (189±63 vs 106±36, t=－5.23), Cdyn-m [(0.16±0.07) vs (0.05±0.03) ml/(kg?cmH 2O), 1 cmH 2O=0.098 kPa; t=－7.29] and Vv/BW increased (22.4±5.0 vs 12.2±3.8, t=－7.46), while S/V (0.23±0.19 vs 0.62±0.38, t=4.10), S/V/BW (15.7±12.4 vs 25.7±17.3, t=2.20), S/V/W/D (0.03±0.03 vs 0.08±0.05, t=3.92) and propensity scores decreased [(12.5±1.2) vs (15.1±1.2) scores, t=7.00]; at 27 d of gestation, Cdyn-m increased [(0.23±0.12) vs (0.16±0.07) ml/(kg?cmH 2O), t=－2.43], but S/V (0.32±0.23 vs 0.57±0.39, t=2.57) and S/V/W/D decreased (0.05±0.04 vs 0.09±0.06, t=2.55); at 28 d of gestation, W/D/BW (270±64 vs 162±33, t=－8.09), LIS-E/BW (72±32 vs 35±20, t=－5.17), S/V (0.90±0.60 vs 0.59±0.48, t=－2.81), S/V/BW (34.0±23.6 vs 15.2±12.7, t=－3.77) and Vv/BW increased (16.9±4.3 vs 9.2±2.9, t=－8.04); the differences were all statistically significant (all P<0.05). (2) Effects of PS: compared with the non-PS group, the PS group had decreased LIS-E/BW at 25, 26 and 27 d of gestation, increased Cdyn-m and Vv/BW at 25 and 27 d of gestation and higher propensity scores at 25 d of gestation (all P<0.05). (3) The correlation between gestational age and each index: gestational age was positively correlated with S/V ( r=0.31, P<0.05), but negatively correlated with W/D/BW and LIS-E/BW ( r=－0.73 and－0.63, both P<0.05). Conclusions:The pharmacological action of prenatal DEX on lung fluid clearance is mainly confined to preterm rabbits at the GA of 28 d which is supported by mechanical ventilation. Prenatal treatment with DEX and/or postnatal PS can improve the early respiratory function in preterm rabbits between GA of 25-27 d, but had no substantial impact on lung fluid clearance. The GA-related lung maturation appears to play a crucial role, in comparison with medications, in lung fluid clearance.
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Objective:To investigate the efficacy of acidified aliphatic ester in the treatment of atopic dermatitis (AD) in mouse models, and to preliminarily explore its mechanisms of action.Methods:Twenty female BALB/c mice aged 6 to 8 weeks were randomly divided into 2 groups: 5 mice in the blank control group were topically treated with absolute ethanol on both ears (14.3 μl per ear) every day, and 15 mice in the model group were topically treated with calcipotriol liniment (14.3 μl per ear) and 20 g/L ovalbumin (25 μl per ear) on both ears every day for 10 consecutive days to establish AD-like mouse models. From day 11, 15 mice in the model group were randomly divided into 3 groups (5 mice in each group), including AD model group, aliphatic ester group, and acidified aliphatic ester group; in the forenoon, all the 3 groups continued to be topically treated with calcipotriol liniment and ovalbumin to maintain AD-like models; in the afternoon, the aliphatic ester group and acidified aliphatic ester group were topically treated with aliphatic ester and acidified aliphatic ester respectively (10 μl per ear), and no treatment was given to the AD model group. Changes in body weight, ear thickness, ear skin lesion scores, and scratching frequency were observed. Ear skin swabs were obtained from the mice on days 10 and 14 for 16S rRNA gene - based microbial diversity tests. On day 14, mice were sacrificed after reflectance confocal microscopy examinations of the ear skin, ear tissues were resected for hematoxylin and eosin staining, mast cell staining, and real-time fluorescence-based quantitative PCR (RT-qPCR), and blood samples were collected for detection of serum IgE levels. One-way analysis of variance was used for analysis of data that met homogeneity of variance criteria, and least significant difference- t test for multiple comparisons. Results:On day 14, the severity of mouse ear lesions was the highest in the AD model group, followed in turn by the aliphatic ester group, acidified aliphatic ester group, and blank control group; compared with the AD model group, the acidified aliphatic ester group showed significantly decreased mouse ear thickness ( F = 897.50, P < 0.001), skin lesion scores ( F = 268.80, P < 0.001), scratching frequency ( F = 64.36, P < 0.001), and epidermal thickness ( F = 256.20, P < 0.001). In addition, RT-qPCR indicated that the expression of inflammatory factors such as interleukin (IL) -33, thymic stromal lymphopoietin, IL-4, and tumor necrosis factor-α in lesional areas, and the degree of mast-cell infiltration were all significantly lower in the acidified aliphatic ester group than in the AD model group ( F = 3.38, 8.70, 41.73, 44.30, 134.30, P = 0.049, = 0.001, < 0.001, < 0.001, <0.001, respectively). Microbial diversity tests showed that the acidified aliphatic ester treatment could inhibit the colonization of Staphylococcus spp. in the ears of AD-like mouse models, and the Shannon index and Simpson index significantly differed among the 4 groups ( F = 9.00, 7.92, P = 0.001, 0.002, respectively) . Conclusion:Acidified aliphatic ester could improve skin lesions of AD-like mouse models, possibly by regulating immunity, suppressing inflammation, and restoring skin microecological diversity.
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Objective:To determine S100A10 protein expression in psoriatic lesions, and to investigate its effect on psoriasis-like skin inflammation in imiquimod (IMQ) -induced mouse models.Methods:From January 2020 to June 2022, skin lesions were surgically collected from 28 patients with psoriasis in the Department of Dermatology, the First Affiliated Hospital of Xinxiang Medical University; normal skin tissues were collected from 18 healthy subjects in the Department of General Surgery and Department of Ophthalmology during the same period, and served as a control group. Immunohistochemical staining was performed to determine the S100A10 protein expression in skin lesions from psoriasis patients and normal skin tissues from healthy controls. Ten wild-type (WT) C57BL/6J female mice and 10 S100A10 -/- C57BL/6J female mice were selected to establish the IMQ-induced mouse models of psoriasis-like skin inflammation. Then, the mice were randomly divided into gene knock-out (KO) /IMQ group, WT/IMQ group, KO/vaseline (VAS) group, and WT/VAS group by using a random number table, and there were 5 mice in each group. The mice in the KO/IMQ group and WT/IMQ group were topically treated with IMQ cream (62.5 mg) on the shaved back daily to establish the mouse models of psoriasis-like skin inflammation, while the mice in the KO/VAS group and WT/VAS group were topically treated with vaseline cream (62.5 mg) daily, and both treatments lasted 7 consecutive days. Skin lesions on the back were observed daily. On day 7, the mice were sacrificed by cervical dislocation, and their dorsal skin tissues were excised. The IMQ-induced psoriasis-like skin inflammation was evaluated by the psoriasis area and severity index (PASI), pathological manifestations of skin lesions were observed by hematoxylin and eosin staining, the expression of S100A10 and Ki-67 in skin lesions was determined by immunohistochemical staining, the expression of STAT3, IL-17A and other cytokines was determined by Western blot analysis, and IL-17 mRNA expression was determined by real-time fluorescence-based quantitative PCR (qPCR). Statistical analysis was carried out by using the independent sample t-test, nonparametric U test, and chi-square test. Results:Immunohistochemical staining showed that the S100A10 protein expression was significantly lower in the psoriasis vulgaris lesions than in the normal control skin tissues ( Z = -3.47, P < 0.001). In the mouse models, the S100A10 protein expression was significantly lower in the skin lesions of mice in the WT/IMQ group than in the skin tissues of mice in the WT/VAS group ( t = 3.64, P = 0.007), and the Ki-67 expression was significantly higher in the KO/IMQ group than in the WT/IMQ group ( t = 2.97, P = 0.041). Additionally, the mice in the KO/IMQ group presented with more severe clinical manifestations such as scales and infiltration compared with those in the WT/IMQ group. Western blot analysis showed that the phosphorylated STAT3/STAT3 expression and IL-17A protein expression was significantly higher in the KO/IMQ group than in the WT/IMQ group ( t = 3.27, 3.48, P = 0.031, 0.025, respectively), and qPCR revealed that the IL-17A mRNA expression was also significantly higher in the KO/IMQ group than in the WT/IMQ group ( t = 2.73, P = 0.029) . Conclusion:S100A10 protein was underexpressed in the skin lesions of psoriasis patients, and the deletion of S100A10 protein aggravated IMQ-induced psoriasis-like skin inflammation in mice, possibly by upregulating STAT3 phosphorylation in the epidermis.
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Objective:To investigate the feasibility of construction of rat models of psoriasis-like lesions by using interleukin (IL) -23/T-helper 17 (Th17) axis-related cytokines combined with imiquimod.Methods:A total of 110 Wistar rats were randomly divided into normal control group, imiquimod alone group, imiquimod combined with interferon (IFN) -α2a (180 000, 60 000, 20 000 IU/kg) groups, imiquimod combined with tumor necrosis factor (TNF) -α (45 000, 15 000, 5 000 IU/kg) groups, and imiquimod combined with IL-2 (90 000, 30 000, 10 000 IU/kg) groups, and there were 10 rats in each group. After hair removal from the central area (2 cm × 2 cm) of the rat back, rats in the imiquimod alone group were topically treated with imiquimod 5% cream at a dose of 20 mg/cm 2 on the shaved back; rats in the imiquimod combined with different cytokine groups were treated with topical imiquimod 5% cream at the same dose on the shaved back for 15 minutes followed by intraperitoneal injections of cytokines at corresponding doses once a day for 10 consecutive days. During the treatment, skin lesions on the rat back were evaluated by using the psoriasis area and severity index (PASI) scores every day. On day 10, serum samples were collected from the rats after anesthesia, and enzyme-linked immunosorbent assay (ELISA) was performed to detect levels of IL-17A, TNF-α, IL-23, IFN-α and IL-1β in the serum samples in each group; then, the rats were sacrificed, lesional skin tissues on the rat back were taken for histopathological examinations and evaluated by Baker scores; an immunohistochemical study was conducted to determine the expression of CD4 and CD8 in some skin lesions. One-way analysis of variance was used for comparisons among multiple groups, and least significant difference (LSD) - t test for multiple comparisons; for data with heterogeneous variance, the Kruskal-Wallis H test was used. Results:On day 3 after molding, the rats in the imiquimod alone group and combination groups gradually presented with psoriasis-like skin manifestations, such as erythema, scales and epidermal thickening; the PASI scores reached a peak on day 7 in the imiquimod alone group, and on day 6 in the combination groups. On day 10, histopathological examination of the skin lesions in the imiquimod alone group and combination groups both showed different psoriasis-like pathological features, such as hyperkeratosis, parakeratosis, acanthosis, thinning or disappearance of the granular layer. There were significant differences in the PASI scores and Baker scores among the normal control group, imiquimod alone group and combination groups ( H = 43.33, F = 42.15, both P < 0.001). The PASI scores were higher in the imiquimod combined with IFN-α2a (180 000 IU/kg) group and the imiquimod combined with IL-2 (90 000 IU/kg) group (9.4 ± 1.1, 8.8 ± 0.6, respectively) than in the imiquimod alone group (7.5 ± 1.1, P = 0.002, 0.030 respectively) ; the Baker scores were higher in the imiquimod combined with IFN-α2a (180 000, 60 000 IU/kg) groups, the imiquimod combined with TNF-α (45 000 IU/kg) group, and the imiquimod combined with IL-2 (90 000 IU/kg) group than in the imiquimod alone group (all P < 0.05). The serum levels of TNF-α, IL-17A, IL-1 β and IL-23 significantly differed among groups ( F = 128.97, F = 6.90, H = 27.45, H = 21.10, all P < 0.05). Compared with the imiquimod alone group, the IL-17A level significantly increased in the imiquimod combined with IL-2 (30 000 IU/kg) group (5.54 ± 1.78 pg/ml vs. 4.20 ± 1.14 pg/ml, P = 0.009), and the IL-23 level significantly increased in the imiquimod combined with IL-2 (90 000 IU/kg) group (37.89 ± 32.85 pg/ml vs. 8.56 ± 6.08 pg/ml, P = 0.036). Immunohistochemical study showed significant differences in the expression of CD4 and CD8 in skin lesions among all groups ( F = 7.21, H = 18.32, both P < 0.001), and the expression of CD4 and CD8 in skin lesions was significantly higher in the imiquimod combined with IL-2 (90 000 IU/kg) group than in the imiquimod alone group ( t = -2.46, -2.32, respectively, both P < 0.05) . Conclusion:Imiquimod combined with IFN-α2a or IL-2 could promote the occurrence of psoriasis-like skin lesions in rats, aggravate the development of psoriasis and prolong the maintenance time of the rat models.
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Animal models play a critical role in the research on the pathogenesis and treatment of white matter injury in premature infants. Rodent modeling is often used to mimic the pathological manifestations of white matter injury in premature human infants. Currently, the most used models include the common carotid artery occlusion combined with the hypoxia model, prenatal/postpartum infection model, chronic hypoxia model, hyperoxia exposure model, neuronal excitotoxicity model, transgenic model, etc. This article reviews the modeling methods, advantages, and disadvantages of the above models.
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ABSTRACT Objective This study verified the replication efficiency of the Rocio virus in a primary culture of mouse neural cells. Methods Mixed primary cultures (neurons/glia) obtained from the brains of newborn isogenic BALB/c mice were inoculated with Rocio virus on the 7 th day of culture, and the development of cytopathogenic effects was monitored. The infection was confirmed via immunocytochemistry (anti-ROCV), while viral replication was quantified in infected primary cultures. The titration method used depended on the infection period. Results Rocio virus efficiently infected primary cultured neural cells, with the highest viral titer causing cytopathic changes was observed at 2 days post infection. The virus-infected primary culture survived for up to 7 days post infection, and viral load quantitation showed viral replication kinetics compatible with the cell death kinetics of cultures. Conclusion The findings of this study suggest that mouse neural cell primary cultures support Rocio virus replication and could be used as an alternative system for studying Flavivirus infection in the central nervous system.