Abstract: Objective To establish xenotransplant paclitaxel-resistant models with A549 human lung cancer cells for the detection of drug resistance. Methods A549-taxol cells (5×10⁶ / mL) were injected into the lungs of nude mice. After 3 weeks, survival and tumor growth were recorded. MTT assays were performed to identify the drug-resistant index and drug susceptibility. Real-time PCR and western blotting were undertaken to detect Pgp170 and MMP-7 mRNA and protein expression, respectively. Results GST-π, P-gp170, and MMP-7 expression were significantly increased (P< 0. 001), and the invasiveness of GST-π-treated A549-taxol lung cancer cells was significantly higher than that of untreated A549- taxol cells and the A549-control group. The survival rate and tumor formation of A549-taxol cells were 100% and 85%, respectively. Immunohistochemistry showed that the drug resistance protein was significantly expressed in the A549-taxol group, and was higher than that in A549-control nude mice. Conclusions GST-π, P-gp170, and MMP-7 expression was associated with paclitaxel resistance in lung cancer. Lung orthotopic transplantation has been gradually established, and the current animal model of paclitaxel-resistant lung cancer remained stable, indicating its usefulness for future experiments.

Abstract: Objective Evidence indicate that the gut microbiota of Parkinson’ s disease patients is imbalanced, but whether the Prnp-SNCA-A53T Parkinson’ s disease transgenic mouse model also has gut microbiota imbalances is unknown. This study aimed to analyze the ecological characteristics of the gut microbiota of this mouse model. Methods Illumina high-throughput sequencing technology was performed to sequence and analyze the biological information of the 16S rRNA gene V3 – V4 region of the fecal microbiota in seven female transgenic mice and 13 wild-type mice of the same sex and age. PICRUST (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) was used to predict the differential function pathways. Results Compared with the wild-type group, the gut microbial alpha diversity of mice in the A53T group had a tendency to increase, and there were also significant differences in microbial composition and species. At the phylum level, the Actinobacteria were increased (P= 0. 0094), and the Bacteroidetes were decreased (P= 0. 0498). At the class level, the Coriobacteriia were increased (P< 0. 0001), and the Bacteroidia were decreased (P= 0. 0398). At the order level, the Coriobacteriales were increased (P<0. 0001), while the Bacteroidales (P= 0. 0398) and the Rhodobacterales were decreased ( P= 0. 0185). At the family level, the Coriobacteriaceae were increased ( P< 0. 0001), while the Bacteroidaceae (P= 0. 0277) and the Rhodobacteraceae were decreased (P= 0. 0185). At the genus level, the Eggerthella were increased ( P= 0. 0002), while the Bacteroides ( P= 0. 0277) and the Rhodobacter were decreased (P= 0. 0249). In addition, there were significant differences between the A53T group and the wild-type group in nine functional pathways, including G protein-coupled receptor, steroid hormone biosynthesis, penicillin and cephalosporin biosynthesis, ubiquinone and other terpenoid-quinone biosynthesis, toluene degradation, glycan biosynthesis and metabolism, electron transfer carriers, meiosis – yeast, and African trypanosomiasis. Conclusions This study indicated that there are imbalances in the gut microbiota of transgenic mice, as well as differences in metabolic pathways between the transgenic mice and the wild-type mice.

Abstract: Objective To construct an acute lung injury mouse model induced by cholic acid with intratracheal instillation or intranasal infusion, and to screen the perfusion method and verify the feasibility of bile acid-induced lung injury. Methods Mice were grouped according to the factorial design of administration method × drug ( 3 × 3). The administration method included tracheotomy, nasal drops for 1 day, and nasal drops for 6 days. Animals were administered either cholic acid, DMSO, or PBS; in total, there were nine groups of mice. During the administration period, body weight changes were monitored. After administration, chest X-ray examination was performed, histological observation of the pathological changes of the lung tissue was carried out, arterial blood was taken to analyze the partial pressure of oxygen (PO₂), and ELISA was used to detect tumor necrosis in the lung tissue, as demonstrated by tumor necrosis factor-α (TNF-α) and interleukin 1β ( IL-1β) expression. Results The X-ray result of the intratracheal instillation and reperfusion cholic acid group showed diffuse infiltration of lung tissue. Gross lung samples showed obvious hemorrhage, while histopathology showed a large amount of inflammatory cell infiltration and alveolar wall thickening. Blood oxygen partial pressure was decreased, and TNF-α and IL-1β were significantly higher than in the other groups. The indexes of the cholic acid group after 1 day of intranasal infusion and 6 days of intranasal infusion were not as good as those in the cholic acid group after intratracheal instillation and reperfusion. Conclusions The acute lung injury model can be successfully constructed by intratracheal instillation of cholic acid.

Abstract: Objective To establish a rabbit model of carotid atherosclerotic plaque by puncturing from central artery of rabbit ear and high-fat diet. Methods All rabbit were fed with high-fat diet for 4 weeks. By puncturing from central artery of rabbit ear, the balloon was inflated to injury intima of carotied artery and then followed with 8 weeks of high-fat diet. The right common carotid artery which damaged by balloon catheter were defined as experimental group and the left common carotid artery as self-control group. MRI and histopathology were used to evaluate common cervical artery lesions in rabbits. Results The success rate of modeling was 85. 0%. On MRI scan image, the wall of carotid artery in the experimental group was thickened and obviously strengthened, while there was no thickening of carotid artery wall in the control group, and no obvious enhancement was observed. In the experimental group, the intima of the common carotid artery wall presented the pathological manifestations of atherosclerotic plaques. In the control group, the inner membrane of carotid artery wall was normal. Conclusions A rabbit model of carotid atherosclerosis was successfully constructed with balloon injury by puncturing from central artery of rabbit ear accompanied high-fat diet. The modified method have advantages of small trauma, simple operation, strong repeatability, short modeling time and high success rate.

Abstract: Objective To establish a cytotoxic T-lymphocyte-associated protein 4 ( CTLA4) humanized mice model, and to provide an effective mouse model for CTLA4-targeted cancer therapeutic antibody research, development, and screening. Methods The animal models were established by CRISPR/ Cas9 and then analyzed by polymerase chain reaction ( PCR), reverse transcription-PCR ( RT-PCR), western blotting, hematoxylin-eosin ( HE ) staining and fluorescence-activated cell sorting. The mouse melanoma cell line ( B16 ) was injected subcutaneously into CTLA4 humanized mice to observe the anti-tumor effects of intraperitoneal injection of the CTLA4 monoclonal antibody, ipilimumab. Results CTLA4-humanized mice could stably express human CTLA4 but not murine CTLA4. CTLA4- humanized mice survived normally within 6 months after birth, and there were no obvious abnormalities in histopathology and the immune system. In CTLA4-humanized mice, ipilimumab slowed tumor growth. CTLA4-knockout mice did not express CTLA4 and died of autoimmune diseases within 3 ~ 5 weeks after birth. Conclusions CTLA4-humanized and CTLA4-knockout mice were established. CTLA4-humanized mice can be used as an animal model for therapeutic antibody- and CTLA4 gene-related drug experiments.

Abstract: Objective To establish a HFMD BALB/ c sulking mouse model infected by coxsackievirus A16, and evaluate immunological and pathological characteristics of this model. Methods In this study, 3-day-old BALB/ c suckling mice were infected with 100 μL coxsackievirus A16 via intraperitoneal injection. The survival rate, clinical score and the 50% lethal dose (LD₅₀ ) were monitored daily 14 days after infection. Challenge with the coxsackievirus A16 with a dose of 3LD₅₀ , 3-day-old BALB/ c suckling mice were monitored daily the survival rate, clinical score and weight within 14 d. 6 d after infection, the concentration of MCP-1, MIP-1 and G-CSF in serum was determined by cytometric bead array (CBA) method ; viral loads of the hind limb muscle, heart, brain, intestine were determined by the real-time RT-PCR; histological change of muscle and brain were determined. Results After different concentrations of virus infection, 3-day- old BALB/ c suckling mice appeared inactivity, hind limb weakness, paralysis and even death, and the virulence of coxsackievirus A16 for 3-day-old BALB/ c suckling mice was 56 LD₅₀ / mL. Within 14 days after infection, compared with the control group, there were significant differences in the survival days, mortality, clinical score, and body weight of suckling mice in the model group (P< 0. 01).6 d post infection, the concentrations of MCP-1 and G-CSF in serum are significantly increased (P< 0. 01) compared with the control group. The viral loads of hindlimb muscle, heart, brain and intestine was significantly higher than that of the control group (P< 0. 01), and the viral loads of hindlimb muscle was the highest. Pathological change showed that there were large area of atrophy and inflammation in the hind limbs muscle and atrophy in the nerve cells of the brain. Conclusions A CoxA16 infected 3-day-old BALB/ c suckling mouse model was successfully established, which can serve for medicine evaluation and mechanism.

Abstract: Objective The method of chronic unpredictable moderate stress combined with rapid and chronic improvement of multi platform water environment sleep deprivation was used to duplicate the rats model of depressive insomnia. Methods 84 Sprague Dawley rats were randomly divided into blank control group, environmental control group, chronic unpredictable moderate stress group (CUMS), 72 h sleep deprivation group ( 72 h SD), chronic unpredictable moderate stress + 72 h sleep deprivation group (CUMS + 72 h SD), 21 days sleep deprivation group (21 d SD), chronic unpredictable moderate stress + 21 days sleep deprivation group (CUMS + 21 d SD), 7 groups, 12 in each group. Weight measurement and food consumption statistics were conducted weekly. Behavioral tests in rats after 14 days of stress modeling and the end of compound modeling were carried out to test the depression and insomnia like behavior of rats. The contents of CRH, ACTH, CORT, Glu, GABA in hypothalamus were detected by ELISA. The contents of NE, 5-HT and DA in hypothalamus of rats were determined by HPLC-ECD. HE staining was used to observe the pathological changes of hypothalamus. Results Compared with the blank control group, CUMS + 72 h SD group and CUMS + 21 d group showed a significant decrease in body weight growth rate and food intake, a significant decrease in the number of autonomous activities, an increase in fecal particles, a significant increase in immobility time in forced swimming, a significant decrease in sugar water consumption, an increase in sleep latency, and a significant reduction in sleep duration, especially in CUMS + 21 d. The contents of CRH, ACTH and CORT in serum, Glu and GABA in hypothalamus were abnormal, contents of monoamine transmitters in hypothalamus were significantly decreased, the arrangement of neurons in hypothalamus was disordered, the gap was enlarged, and the vacuole shape was significant. Conclusions The rat model of depressive insomnia can be duplicated stably by using the method of chronic unpredictable moderate stress combined with sleep deprivation of multi platform water environment for 21 consecutive days.

Abstract: Objective To explore the relieving effect of electroacupuncture on the anxiety behaviors of pain- aversion rats and its effects on N-methyl-D-aspartate (NMDA) and γ-aminobutyric acid (GABA) receptors in the anterior cingulate cortex (ACC). Methods Healthy rats were randomly divided into four groups: control group ( NS group), model group (C-CPA group), normal electroacupuncture group (EA1 group), and pre-electroacupuncture group (EA2 group). The rats of the C-CPA, EA1, and EA2 groups were established pain-aversion models that were injected subcutaneously in the hind paw with complete Freund’ s adjuvant (CFA) and were acclimatized to one side of the cage apparatus to induce conditioned place avoidance (CPA) responses. The rats in the EA1 group underwent intervention with electroacupuncture after modeling, while those in the EA2 group underwent intervention 3 days before modeling. The paw withdrawal thresholds ( PWTs) and anxiety behaviors of rats were observed. Western blotting was used to detect the expression of GABA receptor ( GABAAR, GABABR) and NMDA receptor ( NR2A, NR2B) in the ACC. Results Compared with the NS group, the CPA scores of the C-CPA, EA1, and EA2 groups were decreased (P< 0. 01), and there was no significant difference among the groups, which indicated that the establishment of the pain aversion rat model was successful. Moreover, there was no significant changes in PWTs before modeling. After modeling, rats in the C-CPA group showed hyperalgesia (P< 0. 01) and pain-related anxiety behaviors (P< 0. 01). PWTs and pain-related anxiety behaviors of the rats in the EA1 and EA2 groups were significantly relieved (P< 0. 01 and P< 0. 05, respectively), and there were no significant differences between the two groups (P> 0. 05). Compared with the NS group, GABAAR protein expression in the ipsilateral ACC of the C-CPA, EA1, and EA2 groups was significantly increased (P< 0. 05). There was no significant difference in GABAAR expression in the ipsilateral ACC between the C-CPA and EA1 groups (P> 0. 05), but GABAAR expression in the ipsilateral ACC of the EA2 group was decreased compared with that of the C-CPA group (P< 0. 05). There was no significant difference between the EA1 and EA2 groups (P> 0. 05). GABABR, NR2A, and NR2B expression in the ipsilateral ACC exhibited no differences among the NS, C-CPA, EA1, and EA2 groups (P> 0. 05). Conclusions Both normal- and pre-electroacupuncture intervention could relieve the pain and pain-related anxiety behaviors of C-CPA pain aversion models. The efficacy of pre-electroacupuncture intervention may be related to inhibition of GABAAR expression in the ipsilateral ACC.

Abstract: Objective To systematically evaluate the effect of differentially expressed placental proteins on the growth and development of placenta in rat models with gestational diabetes mellitus (GDM). Methods Databases including CBM, CNKI, Cochrane Library, EMBase, Ovid / LWW, PubMed, Scopus, VIP, WanFang Data and Web of Science were searched by computer and relevant references were manually searched from inception to September 30, 2020. Two researchers independently screened literature, extracted data, assessed quality, and analyzed the outcomes. Results of sixteen studies were included in the analysis. (1) Differentially expressed placental proteins affected the proliferation and apoptosis of trophoblast cells, interfered with placental angiogenesis, increased vascular permeability, enhanced placental oxidative stress and inflammatory responses, and caused abnormal growth and development of placenta in rat models with GDM. (2) Differentially expressed placental proteins were mainly localized in the cytoplasm and / or nuclei of spongiotrophoblasts, labyrinth trophoblasts, giant cells and fetal vascular endothelial cells in rat placenta. (3) Pathological changes included increased trophoblast area, decreased blood vessel density and disordered tissue structure in the placenta of rats with GDM. Conclusions Differential expression of placental proteins may change placental structure and function to cause abnormal growth and development of placenta and thereby promote or aggravate the occurrence of GDM and its complications in rats. However, many high-quality studies are still needed for further verification, because of the limitations of included studies in terms of rigorous experimental design and internal authenticity.

Abstract: Objective To establish an animal model suitable for evaluating the efficacy of an anti-damp-heat (DH) ulcerative colitis compound in Chinese medicine and for new drug development, and to explore the difference and connection between ordinary ulcerative colitis (UC) and UC with DH syndrome. Methods Twenty-four SD rats were randomly divided into the blank control group, UC group and UC+DH group. Rats in the UC group were induced by 5% dextran sodium sulfate (DSS), and the UC+DH group was induced by “ high-fat and high-sugar diet + alcohol + hunger and satiety alternately + high temperature and humidity environment + 5% DSS” for 29 d. The general condition of rats was observed every day for the disease activity index(DAI) and Geboes colonic histopathological scores. Serum levels of ALP, ALT, AST, CHOL, GLU, HDL-C, LDL-C, LDH and TG, and colonic activity of GSH, MDA and MPO were detected. Serum cortisol and colon expression of HSPs, IL-10, MIP-1α, MIP-1β and TNF-α were detected by ELISA. Results The two models exhibited different degrees of intestinal inflammation, increased DAI and Geboes scores, and different degrees of damage to liver function. Compared with the UC group, pathological changes were more severe in the UC+DH group. In addition, serum levels of TG, LDL-C, ALP, ALT, AST, CHOL, cortisol, DAO, HSPs, MDA, MPO, MIP-1α, MIP-1β and TNF-α, and colonic levels of GSH and IL-10 were significantly increased in the UC+DH group, whereas the serum concentration of HDL-C was significantly decreased. Conclusions Intestinal mucosal injury and liver injury were found in both rat UC models. However, the hot and humid environment aggravated the degree of intestinal mucosal injury induced by DSS in UC rats, and its mechanism may be related to promoting inflammation, lipid peroxidation, and intestinal mucosal permeability.

Abstract: Objective To establish a rat model of chronic hyperuricemia renal damage, and to provide a model tool for the development of drugs for anti-chronic hyperuricemia renal damage. Methods Forty male sprague-dawley (SD) rats were randomly divided into five groups: normal group, group A (fed with 2% oteracil potassium + 12% yeast extract + 86% common feed), group B ( fed with 0. 15% adenine + 10% yeast extract + 89. 85% common feed), group C (100 mg / kg adenine + 1500 mg / kg oteracil potassium, intragastric administration in the morning and evening), and group D (50 mg / kg adenine + 1500 mg / kg oteracil potassium, intragastric administration every morning). The observation time was 5 weeks. The body weight, serum uric acid, creatinine, urea nitrogen, and other indicators of rats in each group were detected every week. After 5 weeks, the ratio of bilateral kidney weight to body weight as well as a pathological section of kidney were observed. Results The weight of rats decreased, the ratio of kidney weight to body weight increased, serum creatinine and urea nitrogen all increased, kidney color changed significantly, and kidney damage was identified by hematoxylin-eosin (HE) staining and urate staining in group C compared with the normal group. Conclusions In conclusion , intragastric administration of 100 mg / kg adenine + 1500 mg / kg oteracil potassium in the morning and evening was the best way to establish a rat model of chronic hyperuricemia renal damage.

Abstract:Blood stasis syndrome is one of the common syndromes in traditional Chinese medicine, and it often appears mixed with other syndromes. The diagnosis and treatment of this syndrome is significant in traditional medicine. With increasing investigation of the blood stasis syndrome in modern medicine, it is important to establish more reliable animal models of this syndrome. The purpose of this paper is to review the research progress using blood stasis syndrome animal models in recent years and to provide a reference for the future application and development of blood stasis syndrome models.

Abstract:Alzheimer’ s disease (AD) is an age-related progressive neurodegenerative disease of the brain. The main clinical feature is cognitive dysfunction. As the global population ages, AD has become a major focus of domestic and international research. Appropriate animal models are of great significance for experimental research on disease mechanisms and drug screening for AD. The AD animal models include natural aging and transgenic models, as well as physically and chemically induced AD models. In this article, we compare and summarize the commonly used animal models in AD research and provide a reference for the selection of appropriate animal models in future studies of AD.

Abstract:In recent years, owing to the increasing incidence of intervertebral disc degeneration and age-related early onset, spinal diseases caused by intervertebral disc degeneration have reduced quality of life. The uncertainty of the mechanism of degeneration increases the complexity of the treatment method . To further explore the causes of intervertebral disc degeneration and its pathology, we conducted a literature review to explore the differences between animal models of intervertebral disc degeneration to enable the optimal simulation of human intervertebral disc degeneration to be identified.

Abstract:Uterine fibroids are benign tumors formed by hyperplasia of uterine smooth muscle tissue, and are the most common tumors in women, but their exact cause has not yet been determined. Hysteromyoma is a common clinical feature of hysterectomy. However, hysterectomy can seriously impair fertility. For asymptomatic women who wish to maintain their reproductive health, medication remains the optimal choice. Therefore, animal models are necessary to explore the pathogenesis of fibroids and to develop effective drugs for clinical application.

Abstract:Parkinson’ s disease ( PD) is a regressive disease of the central nervous system. The movement symptoms of PD mainly includes resting tremor, muscle rigidity, bradykinesia, and postural instability. Because of the lack of research on the human brain and the absence of equivalent spontaneous diseases in animals, the development of rational scientific animal models will provide essential research tools in basic and clinical research into PD. Such models may help us to reveal the pathogenesis of PD, and to develop new treatment strategies and drugs.

Abstract:Traditional Chinese medicine (TCM) syndrome models have only recently been explored. To promote the modernization of TCM, in-depth research, and discussion of TCM syndrome, animal models are required. As a common syndrome type of cardiovascular disease, the establishment of an animal model of qi deficiency and blood stasis syndrome is a hot issue in the experimental study of TCM syndrome. Through research on various animal models of heart qi deficiency and blood stasis syndrome, the macroscopic signs, echocardiography, hemodynamics, and other indicators of experimental animals were observed, and the modeling method were Objective ly evaluated. At the same time, the current animal model of TCM syndrome was proposed. The existing problems in the study and prospects of heart qi deficiency and blood stasis syndrome clinical treatment could be addressed by the development of new models.

Abstract:Non-human mammalian cell lines play an increasingly important role in medical and biological research. Like human cells, non-human mammalian cells face the problems of misuse, pollution and variation. However, compared with human cells, the authentication of non-human mammalian cell lines has received less attention and has not yet been fully explored. To date, research into the authentication of non-human mammalian cell lines has mainly focused on cell lines from mice, Chinese hamsters, and African green monkeys. The authentication method mainly use short tandem repeat (STR) markers, supplemented by single nucleotide polymorphism (SNP) analysis. Here, we review the development of the authentication of non-human mammalian cell lines in different species.