Abstract:Objective To determine the effect of imiquimod on psoriasis?like lesions in different strains of mice and the mechanism involved. Methods Two mouse strains, BALB/ c and C57BL/6, were chosen, onto the back of which 62. 5 mg of 5% imiquimod cream was applied. PAIS score was used to determine the morphological changes of mouse back skin. The histopathological changes of skin were observed by HE staining, the thickness of the epidermis was measured under a microscope, and the relative expression of cytokines related to the IL?23/ IL?17Axis was measured by real?time PCR. Results With the passage of time, psoriasis?like skin lesions gradually formed on the back skin of both BALB/ c and C57BL/6 mice. Compared with those in C57BL/6 mice, the PAIS score of BALB/ c mice was higher, and histopathological changes and epidermal thickening ( P < 0. 05 or P < 0. 01) were also more significant. Two days after the application of imiquimod, the relative expression of IL?23, IL?17A, and IL?17F mRNA in the two mouse groups all increased significantly, and both groups showed a dynamic change of an initial increase and then a decrease. The peak of expression in BALB/ c mice appeared 2 days after the application, and in C57BL/6 mice it appeared at 4 days. Conclusions Imiquimod can induce psoriasis?like lesions in both BALB/ c mice and C57BL/6 mice, but the lesions in BALB/ c mice appeared to be more typical, potentially making them more suitable for human psoriasis research. The activation of the IL?23/ IL?17Axis is one of the most important inducers of skin lesions.

Abstract:Objective To analyze the structure and composition of the tree shrew hindgut microbiota by Illumina PE250 sequencing technology. Methods Three male 10?month?old tree shrews were selected. The V3 - V4 region was amplified with universal primers of bacteria 16SrDNA and sequenced with the Illumina PE250 platform after the extraction of total DNA. Results A total of 160 157 valid sequences and 437 operational taxonomic units (OTUs) were obtained. The bacteria in the tree shrew hindgut are from 9 phyla, 19 classes, 37 orders, 68 families, 137 genera, and 194 species. Among them, (i) Firmicutes and Proteobacteria had the highest abundance, at 63. 30% and 28. 52%, respectively. (ii) The dominant classes were Bacilli and Gammaproteobacteria ( 51. 46%, 28. 02%). ( iii ) The abundances of Lactobacillales and Enterobacteriales were high, at 45. 33% and 27. 51%. (iv) Dominant families were Lactobacillaceae (35. 30%) and Enterobacteriaceae (27. 51%). (v) Lactobacillus and Escherichia?Shigella were the dominant genera (35. 30%, 27. 49%). (vi) At the species level, the abundance of cultured bacteria was 37. 10%, that of unclassified bacteria was 27. 69%, and that of uncultured bacteria was 35. 21%. Lactobacillus salivarius was the most abundant species (26. 66%) among the cultured bacteria. Of the 20 species with the highest abundance, eight (40%) were unclassified and new species. Some differences among individuals in terms of bacterial communities were identified. Conclusions The tree shrew hindgut microbiota has rich diversity, and there are many bacteria that have not been identified and have relatively high abundance, which needs further study.

Abstract:Objective To investigate the variations of blood biochemical and physiological characteristics in the process of breeding inbred diabetic Mongolian gerbils. Methods A total of 17 blood biochemical and 22 physiological indexes of blood samples from F9 to F14 progenies were tested using an automatic biochemical analyzer and automatic blood cell analyzer. Results The results showed that uric acid (URIC), lactate dehydrogenase (LD), creatine kinase (CK), amylase (AMY), glucose (GLU), percentage of mononuclear cells (MON%), mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration (MCHC), and mean platelet volume (MPV) increased with an increasing number of generations. Meanwhile, cholesterol ( CHOL), eosinophil direct count ( EOS), percentage of eosinophils ( EOS%), red blood cells ( RBC), hematocrit ( HCT), and hemoglobin ( HGB) decreased with the generations. The levels of albumin (ALB), total protein (TP), triglyceride (TG), high?density lipoprotein (HDL), urea nitrogen (BUN), total bilirubin (TBIL), creatinine (CRE), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and the other indexes remained stable. Conclusions The blood biochemical and physiological indexes vary across the generations in the process of inbreeding diabetic gerbils, especially for those indexes related to glucose and lipid metabolism, which are indicators of the pathogenic characteristics of diabetes. The findings also imply that selective inbreeding exerts an influence on the animals’ biological characteristics.

Abstract:Objective To screen the polymorphisms of the promoter regulatory region of the PPARγ2 gene in Wuzhishan minipigs. Methods Five pairs of primers were designed based on the promoter regulatory region of the PPARγ2 gene in pigs. Then, the polymorphisms of 57 outbred Wuzhishan minipigs were detected; moreover, the promoters, transcription factor?binding sites, and secondary structures of RNA and CpG islands of the regulatory region were predicted and analyzed by various types of bioinformatic software. Results Nine single?nucleotide polymorphisms (SNPs) were found, which were - 1595T/ C, - 1534G/ A, - 1262C/ A, - 1220C/ A, - 1017A/ G, - 963A/ G, -955G/ A, -866A/ G, and - 333G/ A. SNPs were not located in the region of promoters that were identified by the software, but the -333G/ A mutation was located in the core promoter region ( - 656 to - 23 bp). Transcription factor?binding sites were altered near the mutational locus. The minimum free energy of secondary structures of RNA was changed and the structure of RNA was significantly altered. CpG islands were not found in the target sequence. Conclusions The screened SNPs of the promoter region of the PPARγ2 gene in Wuzhishan minipigs may affect regulation of the expression of this gene, which lays the foundation for further study of its function.

Abstract:Objective To explore the effect of compound Ejiao slurry on mouse uterus. Methods Female mice were randomly divided into four groups: sham operation group, sham operation with Chinese medicine group, ovariectomized group, and ovariectomized with Chinese medicine group. The sham operation with Chinese medicine group and ovariectomized with Chinese medicine group were treated with compound Ejiao slurry gavage. HE staining, immunohistochemical staining, and Western blot were used to observe and compare the endometrium among the groups. Results Compared with that of the sham operation group, the endometrium of the sham operation with Chinese medicine group was thicker. In addition, the number of glands and the expression of vascular endothelial growth factor?A (VEGF?A) in endometrium slightly increased ( P < 0. 05). However, the differences between the ovariectomized group and ovariectomized with Chinese medicine group were not significant. Conclusions The compound Ejiao slurry can affect the growth of reproductive organs in female mice. It may regulate the endometrium by acting on the ovary.

Abstract:Objective To observe the changes of DR5 and DcR2 expression in ovarian granulosa cells of polycystic ovary syndrome (PCOS) rats, and to explore the possible mechanism of follicular development disorder in these rats. Methods Rats were subcutaneously injected with sodium prasterone sulfate to establish the PCOS model. Immunohistochemical and RT?qPCR analyses were used to observe and compare the expression of DR5 and DcR2 in ovarian granulosa cells of the rats in the PCOS group and a control group. Results Immunohistochemical results showed that, in PCOS rats, in ovarian granulosa cells at each level, the expression of DR5 was significantly higher than that in the control group ( P < 0. 05). In ovarian preantral and antral follicular granulosa cells of the rats in the PCOS group, the expression of DcR2 was significantly increased compared with that in the same follicle level of the control group ( P < 0. 05), while there was no significant difference in DcR2 expression in primordial follicle granulosa cells ( P > 0. 05). RT?qPCR analysis showed that DR5 mRNA and DcR2 mRNA were expressed in the two groups, and their expression in the rat ovarian granulosa cells of the PCOS group was significantly higher than that in the normal control group ( P < 0. 05). Conclusions The abnormal expression of DR5 and DcR2 in granulosa cells may play a role in the abnormal development of follicles in PCOS rats. They may affect the development of follicles by participating in the process of granulosa cell apoptosis.

Abstract:Objective To investigate the possibility of establishing the high?cholesterol forage?feeding tree shrew as a cholesterol?induced gallstone model and to determine the association of substance P (SP) with this model. Methods Upon establishment of the tree shrew cholesterol?induced gallstone model, the SP expression of the digestive tract was detected with immunohistochemistry method and compared among the model groups and the control group. Results In the model groups, immunohistochemistry showed that gastrointestinal SP expression was lower than that in the control group ( P < 0. 05). The tree shrew gallstone model was negatively correlated with SP expression level. Conclusions Tree shrew gallstones are associated with the regulation mechanisms of hormone secretion in the digestive tract.

Abstract:Objective To provide a pathological model for the screening and study of drugs for the treatment of hyperuricemic nephropathy by establishing a reasonable and stable mouse model with hyperuricemic kidney injury. Methods By administering one, two, or three of five drugs, oteracil potassium, hypoxanthine, adenine, ethambutol, and yeast extract, a model was established with hyperuricemia renal injury in KM mice, at different modeling times by using different modeling doses and modeling methods. The changes in serum uric acid, urea nitrogen, creatinine, liver xanthine oxidase (XOD), and adenosine deaminase (ADA) activity were determined, and the variations in weight of each group were analyzed. Results Compared with the normal group, the serum uric acid level and urea nitrogen level in the mice significantly increased ( P < 0. 01) in the group with 1?day combination medication of hypoxanthine and oteracil potassium, the renal tubular type was abundantly observed in renal cortex, and salt crystals were abundantly observed in renal medulla. The serum uric acid level and urea nitrogen level in the mice increased significantly ( P < 0. 01), and liver XOD activity decreased ( P < 0. 05) in the group with 7?day combination medication of hypoxanthine, ethambutol, and oteracil potassium, and eosinophilic insoluble proteins were found in some of the proximal tubules of the renal cortex. The serum uric acid level, urea nitrogen level, and creatinine level in the mice significantly increased ( P < 0. 01) in the group with 14?day combined medication of yeast extract and oteracil potassium and the group with 14?day combined medication of yeast extract, adenine, and oteracil potassium. Renal tubular epithelial cells fell off in the renal cortex, and eosinophilic insoluble proteins were visible in some of the proximal tubules of the mice in the group with combined medication of yeast extract and oteracil potassium. Salt crystals were abundantly observed in the renal medulla of the mice in the group with combined medication of yeast extract, adenine, and oteracil potassium. The body weight of the yeast extract and oteracil potassium group increased faster than that of the yeast extract, adenine, and oteracil potassium group, with a significant difference between their body weights ( P < 0. 05). Conclusions Compared with other modeling method , the mouse model with hyperuricemic renal damage induced by the combined medication of yeast extract and oteracil potassium was more stable and its establishment had no significant effect on the body weight of the mice. Therefore, it is more suitable to use yeast extract combined with oteracil potassium once a day for 14 days to establish a mouse model of hyperuricemia renal damage.

Abstract:Objective To construct an intrahepatic model of advanced liver cancer (hepatocellular carcinoma, colorectal cancer liver metastasis, breast cancer liver metastasis, non?small cell lung cancer liver metastasis) by injecting cancer cells through the hepatic portal vein. Methods MHCC97?H, a highly aggressive cell line of hepatocellular carcinoma; SW480, a metastatic colorectal cancer cell line; MDA?MB?231, a human triple?negative breast cancer cell line; or A549, a non?small cell lung cancer cell line, was cultured and seeded into the liver of BALB/ c nude mice by injecting cancer cells through the hepatic portal vein. After 3 - 4 weeks of growth, mice were analyzed by PET/ CT screening. Mice were injected with approximately 200 μCi nuclide probe18 F?FDG through the tail vein. The BALB/ c nude mice were analyzed by PET/ CT screening after 30 -40 min. The mice were then collected, and their livers were obtained and photographed to determine the presence of tumor nodules on the liver of BALB/ c nude mice. Results MHCC97?H, A549, SW480, or MDA?MB?231 cells grew and multiple diffuse tumor nodules formed in the nude mouse livers. Nodules in the nude mouse livers formed by cancer cells could be identified by PET/ CT screening; the PET/ CT intensity of nodules formed after A549, SW480, or MDA?MB?231 cell injection was weaker than that of MHCC97?H cells. Conclusions An intrahepatic tumor animal model of advanced liver cancer has been successfully constructed by injecting cancer cells through the hepatic portal vein. This is a promising model for research on liver cancer or antitumor drugs.

Abstract:Objective To investigate the relationship between B?lymphocyte?induced maturation protein?1 (Blimp?1) and the proliferation, apoptosis, and invasion of the hepatocellular carcinoma cell line SMMC?7721. Methods The experiment included a control group, shRNA?NC group, and shRNA?Blimp1 group. The expression of the Blimp?1gene was silenced in hepatocellular carcinoma cells. Real?time quantitative PCR (RT?PCR) and Western blotting were used to detect Blimp?1 mRNA and protein expression. Proliferation and apoptosis were measured by the CCK?8 method and flow cytometry. The Transwell method was used to observe the invasion ability of cells. The protein levels of IFN?γ and IL?2 were analyzed by ELISA. Results Compared with the levels in the control group, the expression of Blimp?1 mRNA and protein in the shRNA?Blimp1 group was significantly decreased ( P < 0. 05). Silencing Blimp?1 had significant effects on inhibiting SMMC?7721 cell proliferation ( P < 0. 05), promoting cell apoptosis ( P < 0. 05), significantly reducing the invasive ability of cells ( P < 0. 05), and significantly increasing the level of IFN?γ protein ( P < 0. 05), but had nosignificant effect on IL?2 protein level. Conclusions Silencing Blimp?1 can promote the expression of IFN?γ protein and inhibit the proliferation, invasion, and apoptosis of hepatoma cells.

Abstract:Objective To improve the results of in vitro fertilization (IVF) for non?obese diabetic (NOD) mice. Methods NOD female mice were divided into four age groups: 4, 6, 8, and 10 weeks old. Sperm from 3 -6?month?old male NOD mice were used for in vitro fertilization. The next day, the number of usable embryos per female mouse was counted and the experiment was repeated twice. NOD mouse embryo transfer data collected from 2016 -2018 were used for the comparison of pup rates. Results Twenty?four hours after the first experiment, the means ± SD of the number of two?cell?stage embryos per group were 13. 4 ±2. 5, 22. 7 ±2. 9, 22 ± 7. 0, and 30. 5 ± 10. 9, respectively. Twenty?four hours after the second experiment, the numbers of two?cell?stage embryos per group were 7. 8 ± 2. 6, 16. 5 ± 4. 5, 14. 2 ± 6. 5, and 24. 5 ±5. 9, respectively. The mean ± SD rates of pups born from transferred embryos after in vitro fertilization in 4, 8, and 10 weeks were (35. 32 ±9. 9)%, (36. 19 ±6. 2)%, and (43. 15 ±6. 9)%, respectively. Conclusions Ten?week?old NOD mice respond better to IVF than 4?, 6?, or 8?week?old ones. Therefore, we suggest the selection of 10?week?old NOD female mice for in vitro fertilization.

Abstract:On the basis of investigating and combing the current situation of laboratory animal resources and their supply and marketing in China, this article analyzes the application needs of user groups in laboratory animal industry for information services on laboratory animal resources, and explores the service platform mode for sharing laboratory animal resources in China under the “Internet + ” environment, fully utilizing the optimization and integration role of “Internet + ” in the allocation of social resources, and contributing to the construction and sharing of resources in the laboratory animal industry. This article focuses on the one?stop laboratory animal resource?sharing service platform as an example, and introduces ideas for the design and functional module realization of the platform.

Abstract:Objective To compare the livers of four commonly used laboratory animals (mouse, rat, guinea pig, and rabbit) with the human liver regarding tissue morphology, in order to provide a reference for study of the liver and the choice of laboratory animal. Methods Livers of normal mice, rats, guinea pigs, and rabbits were obtained. The morphology, hepatic lobules, organ coefficient, and nucleoplasm ratio of the livers of the four animals were investigated and compared with those of human. Results The results showed that the morphological characteristics of rabbit liver were the most similar to those of human liver, and the shapes of rabbit liver and human liver were irregular wedge and wedge, respectively. The nucleoplasm ratio of rabbit was also the closest to that of human, at 0. 158 and 0. 149, respectively. The hepatic lobules of guinea pig were the closest to those of human, with both five lobules. The liver coefficient of guinea pig was also the closest to that of human, at 3. 096% and 2. 35%, respectively. Conclusions The organization and morphological characteristics of the liver of rabbit and guinea pig are closest to human, so these two animals may be more suitable for liver?related research.

Abstract:Objective To explore the application of a quality control circle (QCC) for reducing the rate of non?standardized behaviors in animal experiments. Methods A QCC team named “Love and Welfare” was set up. According to the steps of QCC activity, the causes of non?standardized behaviors during animal experiments of the researchers in our hospital were analyzed, and the related countermeasures were formulated and implemented. Results Through this QCC activity, the rate of non?standardized behaviors occurred during animal experiments in our hospital was reduced from 82. 5 times to 22. 33 times per week. Conclusions QCC activity has significant effects on reducing the rate of non?standardized behaviors and can be used for enhancing the quality of animal experiment management.

Abstract:Objective To investigate whether the animal nose?only inhalation exposure device can operate normally and produce aerosol particles of a specific size, to explore the air tightness and uniform distribution of the aerosol during operation, and to examine the disinfection effect of disinfectant on the system. Methods Escherichia coli (E. coli) aerosol and mouse hepatitis virus MHV?JHM aerosol were developed by the nose?only inhalation exposure device, and a solution of 75% ethanol and 3% hydrogen peroxide was run through the nebulizer to disinfect the system. E. coli and MHV?JHM were collected respectively in the exposure chamber, at sites where leaking easily occurred, including after disinfection. The sampled E. coli was cultured. In addition, RNA from the sampled MHV?JHM was extracted and one?step RT?PCR was then used to test the presence of viral RNA from the samples. Results The result showed that the mass median aerodynamic diameter of the E. coli aerosol was 1. 27 ± 0. 61 μm, that colonies grew after coating of a plate with sample liquid collected in the exposure chamber, that the numbers of cultured colonies by NC membrane sampling of different ports were similar, and that no colony growth was found on the sedimentation plate at sites where leaking readily occurred or on the sampling plates after disinfection. There was a positive result for MHV?JHM virus RNA of the sampling solution from the liquid impinger, while there were negative results for MHV?JHM virus RNA of the sampling solutions from the sites where leaking easily occurred and after disinfection. Conclusions The results show that the animal nose?only inhalation exposure device can operate normally and develop an aerosol that is passed through and distributed evenly in each port of the exposure chamber during operation; moreover, the device was shown to have good air tightness. The system can be effectively disinfected by using the corresponding disinfectant, and the disinfection effect is comprehensive.

Abstract:Objective To develop a biosafety negative?pressure poultry isolator. Methods In accordance with national standards for animal isolation equipment, taking into account the actual needs of airtight transfer and disinfection functions, the structure of an isolator was designed. Results The isolator was integrated with an operating system, ventilation and filtration system, transfer system, automatic control system, sterilization connection system, and sewage collection and treatment system. Upon testing by a third party, it was confirmed that its technical indicators meet the national standards. Conclusions The isolator is suitable for feeding and experimental research of animals with highly pathogenic infections, meeting the practical needs of biosafety laboratories in China.

Abstract:Circular RNAs (circRNAs) are a newly discovered class of covalently closed circular endogenous noncoding RNAs that differ from miRNAs and lncRNAs. Many reports have recently been published on the involvement of circRNAs in the mechanism of disease development, providing a new theoretical basis for tumor prevention, diagnosis, and precise treatment. This article reviews the biological characteristics and mechanisms of circRNAs and their role in the development of several common oral squamous cell carcinomas.

Abstract:Rat is a commonly used experimental animal. The open?field behavior test is one of the most commonly used methods to detect emotional behavior in animals, particularly for rats in recent years. The conditions of the open?field behavior test can cause changes in the physiological and psychological state of rats, affecting the interpretation of rat behavior, and thus affecting the determination of the rat model of diseases. To select suitable experimental conditions and achieve reasonable interpretation of behavior and an appropriate model, this article presents a review of the research literature on the open?field behavior test of rats published in the last 5 years.

Abstract:With the improvement of human material living standards and the change of lifestyle associated with modern living, metabolic?related diseases, which affect the health of people globally, have become an important issue. To study the occurrence, development, transformation, and prevention of this kind of disease, it is vital to construct an appropriate animal model. Tree shrews (Scandentia animals) have a closer phylogenetic relationship with primates, which makes the former a useful animal model, compared with the widely used rodent models, for the simulation of disease. At present, animal models including for diabetes, atherosclerosis, nonalcoholic fatty liver, gallstones, and osteoporosis have been established using tree shrews, and their mechanisms and related factors have been studied. This article outlines the progress of research on tree shrews for insights into human metabolic diseases.

Abstract:There is currently no effective treatment for some motor neuron diseases, such as spinal muscular atrophy and amyotrophic lateral sclerosis. With the development of induced pluripotent stem cells and cell reprogramming technology, it is possible to induce the production of motor neurons from a patient’s somatic cells in vitro for autologous transplantation. The differentiated motor neurons can also be used as cell models for studying disease mechanisms and for drug screening. Many researchers are now focusing on the differentiation of motor neurons from fibroblasts in vitro; here, we summarize three different methods for this, namely, differentiation through induced pluripotent stem cells, neural stem cells, and direct conversion.

Abstract:The stellate ganglion (SG) is functionally a sympathetic ganglion. Stellate ganglion block (SGB) refers to a variety of methods for sympathetic blockade of the head, face, neck, upper limbs, and upper chest, which are innervated by stellate ganglia. SGB has been demonstrated to produce therapeutic or protective effects in multiple systems throughout the body, but the exact mechanism involved has not yet been elucidated. Rat SG resembles the anatomy and function of the equivalent system in human, so rat is an ideal model for mimicking human SGB and can provide a basis for researching the molecular biological mechanism behind the effects of SGB. At present, the methods of rat SGB have rarely been described and mostly have been simply mentioned as part of the experimental operation, and there is no literature to summarize the approaches used. Therefore, the method for establishing the rat SGB model is reviewed here.

Abstract:Objective The rational use of antibiotics is critical for the prevention of surgical site infection. Therefore, revealing its relationship with the infection of clean incisions and its impact on the clinical surgical infection rate would provide a scientific basis for the rational use of drugs and clinical interventions during the surgical period. Methods Patients who had undergone cleaning of infected incisions were registered and tracked according to the medical records from an electronic information platform, which allows retrospective and real?time investigation. The following information was recorded and analyzed in an Excal form: name, age, sex, length of hospital stay, operation, amount of bleeding, information on antibacterial drug use, combination drug therapy, nosocomial infection and incision healing, and the database was built. Wound infection was diagnosed based on Hospital Infection Diagnostic Criteria (Trial) and Surgical Site Hospital Infection Prevention and Control Technical Guidelines issued by the Ministry of Health. Finally, the effect of rational use of antibiotics on clinical operation was evaluated according to the results of statistical analysis. Results There were a total of 970 cases registered with incision cleaning in this study. Prophylactic antibiotics were used in 653 cases, with a use rate of 67. 32%. The cases with antibiotics usage in the perioperative period were divided into three groups based on the time distribution of antibiotic treatment: there were 437 cases (66. 92%) with the usage of antibiotics 0. 5 - 2 h before the operation, 190 cases (29. 10%) with the usage after the operation, of which emergency operation accounted for 123 cases, and 26 cases (3. 98%) with the usage 1 day before the surgery. Among the 653 cases, 194 cases (29. 71%)were recorded with antibiotic usage in the perioperative period < 24 h, 164 cases (25. 11%) 24 - 48 h after the operation, 101 cases (15. 47%) 48 - 72 h after the operation, and 194 cases (29. 71%) > 72 h after the operation. There were 516 cases with reasonable indications, at a rate of 79. 02%. Among the total of 970 patients included in the study with incision cleaning in surgery, the number of grade A healing cases was 945, with the grade A healing rate being 97. 42%, leaving 5 cases (0. 52%) suffering from incision infection. Therefore, the infection rate met the national regulations on nosocomial infection rate of clean incisions of ≤ 1. 5%. Conclusions Through a comparative analysis of the rationality of 563 cases with perioperative use of antibacterial drugs, 79. 02% of the cases were reasonable in terms of antibiotic usage during the operation, and the clean incision infection rate was less than 1. 5%, in line with national regulations. However, there is a certain gap between the use of antibiotics in the perioperative period and the requirements of the Ministry of Health. The management and supervision of the rational use of antimicrobial drugs should be further strengthened.