Abstract:Objective The expression pattern of TET family proteins was investigated in a mouse lung cancer model. Methods The mice were divided into three groups (with 10 mice in each group): control group (Con), urethane- induced group (Ure), and HU-treated group (Ure-HU). Urethane ( 800 mg / kg) was injected intraperitoneally for 10 weeks to establish the mouse lung cancer model in the Ure and Ure-HU groups. After 10 weeks, the Ure-HU group was given the drug HU (500 mg / kg) for 21 days. The morphology was observed thereafter, while qPCR and western blotting were used to analyze the expression patterns of TET1, TET2, and TET3 in the three groups. Results Compared with that of the Con group, the weights of the Ure group and Ure-HU group decreased. Morphological observation revealed lung tissue lesions in the Ure and Ure-HU groups. qPCR result suggested that the expression of TETs was decreased in the Ure group(P < 0. 05). After HU treatment, the expression of TET1 and TET2 was increased in the Ure-HU group (P < 0. 05). The data from western blotting were consistent with the qPCR result . Conclusions The expression of TETs was abnormal in a mouse lung cancer model induced by urethane. These result suggest that TET1 and TET2 play important roles in lung cancer.

Abstract:Objective To analyze and evaluate the gait changes of Parkinson’s disease in mice using the CatWalk gait analysis system. Methods A Parkinson’s disease mouse model was established by the intraperitoneal injection of MPTP. After the model had been successfully established, the CatWalk gait analyzer was used to measure the relevant gait parameters, and the result were statistically analyzed. Results Compared with the control group, the pace of the mice in the model group was disordered. The run duration, maximum variation, stance, and step cycle were increased significantly in the MPTP group. However, the average speed, cadence, swing speed, and stride length were significantly reduced in the model group. Conclusions CatWalk can be used as an intuitive, precise, and effective new method to evaluate Parkinson’s mouse models.

Abstract:Objective To explore the mechanism of hepatocarcinogenesis by establishing a C57BL/ 6 chemically induced liver cancer mouse model to simulate the development of liver cancer. Methods A total of 110 C57BL/ 6 mice were randomly divided into a control group ( n = 30) and a model group ( n = 80). The mice in the normal control group were left untreated. The mice in the model group were intraperitoneally injected with DEN followed by the administration of CCl4 orally. The liver morphology was observed, and the levels of alanine aminotransferase ( ALT) and aspartate aminotransferase (AST) in serum were determined by spectrophotometry. HE staining was used to observe pathological changes; the body weight of all groups was also recorded. Results Compared with those in the control group, the body weight of the model group was significantly decreased, as were the levels of ALT and AST. HE staining result showed that the liver of the model group exhibited clear pathological changes. Conclusions Diethylnitrosamine combined with carbon tetrachloride can induce liver cancer in C57BL/ 6 mice, providing a reliable animal model for studying the molecular mechanism of liver cancer development.

Abstract:Objective To observe the plastic changes of pain threshold in myocardial infarction rats, and to investigate the effects of nicotine on visceral pain and whether it has protective effects on ischemic myocardium. Methods Thirty healthy male Sprague-Dawley rats weighing 180 to 200 g were selected and divided into a sham surgical group, model group, or nicotine treatment group at random. The changes of electrocardiogram (ECG), pain threshold ( thermal pain threshold PWL, mechanical pain threshold PWT), left ventricular pressure amplitude, cardiac body mass index, and creatine kinase isoenzyme (CK-MB) were observed in all groups of rats. Results Upon comparing the model group with the sham surgical group, the mechanical pain threshold decreased and the thermal pain threshold became larger in the former group (both P<0. 05), while the plasma CK-MB activity, cardiac body mass index, and left ventricular pressure significantly increased (all P<0. 05). In addition, when the nicotine treatment group was compared with the model group, the mechanical pain threshold increased in the former group, while the thermal pain threshold decreased, as did the plasma CK-MB activity, cardiac body mass index, and left ventricular pressure (all P<0. 05). Conclusions Nicotine can relieve myocardial ischemia symptoms and visceral pain caused by ischemia in rats with myocardial infarction. Its mechanism may be related to activation of the cholinergic anti-inflammatory pathway. Therefore, it can improve the pain and protect the heart of rats.

Abstract:Objective To investigate the effects of ginseng polysaccharides (GSP) on ethanol-induced behavioral sensitization and its possible mechanism. Methods Eighty Kunming mice were randomly divided into five groups: blank, model, GSP low-dose (100 mg / kg), GSP middle-dose (200 mg / kg), and GSP high-dose (400 mg / kg). Except for the blank group, all animals were provided water or ethanol freely. The concentration of ethanol was gradually increased from 3% to 12% by increasing ethanol 3% every four days. After a 4-day conversion period, the initial dose of 3% ethanol was provided to establish a behavioral sensitization model. Ethanol consumption and locomotor activities of each group were measured daily during the experimental period. During the transition period, an open field test (OFT) was carried out. Activation of behavioral sensitization was determined by locomotor test during the expression period. Western blot and real- time PCR were used to detect protein and mRNA expression, respectively, of ERK, p-ERK, and c-fos, in the prefrontal cortex. Results Three doses of GSP significantly reduced the hyperactivity caused by 9% and 12% ethanol during the transition period, and significantly inhibited the increase of locomotor activity and ethanol intake induced by 3% alcohol challenge (day 25). In addition, middle and high doses of GSP could significantly modulate the number of crawls and standing times in the OFT. GSP could significantly inhibit ethanol-induced increases in malondialdehyde levels in the prefrontal cortex, as well as increased superoxide dismutase and catalase activities. Western blot and real-time PCR result showed that GSP could downregulate protein and mRNA overexpression of p-ERK and c-fos in the prefrontal cortex. Conclusions GSP had an inhibitory effect on ethanol-induced behavioral sensitization and ethanol intake in mice, and this effect may be related to the ERK/ c-fos pathway.

Abstract:Objective To study the effects of aerobic exercise combined with curcumin on insulin resistance and liver pathological changes in ob / ob diabetic mice, to determine the effects of curcumin combined with aerobic exercise on the hepatic fatty acid β-oxidation signal pathway, and to explore the specific mechanisms behind these effects. Methods Thirty-two ob / ob diabetic mice were divided into a diabetic control group (ON group), aerobic exercise intervention group (OA group), curcumin intervention group ( OC group), and aerobic exercise + curcumin intervention group ( OAC group), while another eight C57 / BL6J mice served as a normal control group (NC group). During the experiment, aerobic exercise training was performed on OA group and OAC group mice. Curcumin treatment (50 μg / (g·d)) was applied to OC group and OAC group mice. After 8 weeks of intervention, glucose tolerance and insulin resistance of the mice were determined. The pathological changes and fatty acid β-oxidation signaling pathways in the liver were also measured. Results After the experimental intervention, the insulin sensitivity of the OA group, OC group, and OAC group was significantly increased, and the OAC group almost reached the level of the NC group; the serum TC and TG of the OA group, OC group, and OAC group were lower than those of the ON group, and the liver function was significantly improved; the liver fibrosis level of diabetic mice was significantly improved, and these levels of the OA group, OC group, and OAC group were significantly lower than those of the NC group. There was no significant difference between the OAC group and the NC group in the diabetic control group. There was also no significant change in the expression of fatty acid synthesis-related genes in the liver, but the expression of fatty acid β-oxidation-related genes in the OA group, OC group, and OAC group was significantly higher than that in the ON group, and the expression of fatty acid β-oxidation-related genes in the OAC group was significantly higher than that in the OA group and OC group. Western blotting showed that the fatty acid β- oxidation pathway in the liver of the OA group, OC group, and OAC group. The expression of carnitine palmityl transferase I (CPT-I) was significantly higher than that in the ON group. Conclusions Aerobic exercise combined with curcumin can promote the expression of the rate-limiting enzyme CPT-I involved in fatty acid β-oxidation in the liver of diabetic mice, thereby promoting fatty acid β-oxidation to enhance lipid decomposition, improving pathological changes in the liver and enhancing insulin sensitivity. These result may provide new ideas for the treatment of diabetes.

Abstract:Objective To observe the dose-effect relationship of xingnaojing injection in treating coma in beagle dogs. Methods Male beagles were divided into nine groups at random: Control, Model, Naloxone ( 0. 01 mg / kg),Xingnaojing group (dose 1) (0. 215 mL/ kg), Xingnaojing group (dose 2) (0. 43 mL/ kg), Xingnaojing group ( dose 3, single dose) ( 0. 86 mL/ kg), Xingnaojing group ( dose 3, double dose) (( 0. 43 + 0. 43) mL/ kg), Xingnaojing group (dose 4, single dose) ( 1. 29 mL/ kg), and Xingnaojing group ( dose 4, double dose) (( 0. 65 + 0. 65) mL/ kg). Lu mianning was used for anesthesia, fixed in a custom-made hammer, and a model of traumatic brain injury in dogs was established. MRI was performed, intravenous medication for 14 days. The waking time, coma degree, and cortical electroencephalogram were measured. After the last intravenous administration of medication, MRI was performed and blood was taken to determine liver and kidney function, along with routine blood tests and measurements of NSE and S-100B. Cerebrospinal fluid was used to measure Glu, GABA, and His. Histopathological examination was performed on the heart, liver, spleen, lung, kidney, adrenal gland, brain, pituitary, sciatic nerve, and spinal nerve from animals in each group, and some organs were weighed. Results Xingnaojing injection doses 1, 2 (single dose), and 3 (two doses) significantly reduced the waking time of dogs. Xingnaojing doses 1, 2, and 4 significantly reduced the levels of S100B and NSE in serum. Xingnaojing doses 1, 2, 3 (2 doses), 4, and 5 (2 doses) significantly increased the area of brain injury recovery in dogs. The numbers of lymphocytes and eosinophils in xingnaojing injection group 1 and group 2 significantly decreased. Xingnaojing injection dose 1 significantly reduced the number of mononuclear cells in dogs. Xingnaojing injection doses 4 and 5 had no significant effect on canine blood routine findings. There were no significant differences in ALT, AST, TBil, BUN, and CRE between the xingnaojing injection groups. Xingnaojing injection showed no significant difference in histopathology of canine heart, liver, spleen, lung, kidney, and brain. Conclusions Xingnaojing injection clearly ameliorated the coma caused by traumatic brain injury in beagle dogs, while having no clear toxic effects at a high dose.

Abstract:Objective To explore the damaging effect of busulfan on hematopoiesis and immune functions of KK/ upj-Ay / J (KKA^y) mice with type 2 diabetes mellitus. Methods Male KKA^y mice were randomly divided into busulfan treatment and vehicle control groups. The former was intraperitoneally injected with busulfan at a dose of 40 mg / kg, while the latter was injected with an identical volume of 5% dimethyl sulfoxide; B6 mice were grouped and treated in the same way. Fifteen days after administration, all mice were euthanized for peripheral blood count and classification of hematopoietic progenitor cell (HPC), hematopoietic stem cell (HSC), and long-term hematopoietic stem cell (LT-HSC) frequencies. The function of HPCs was assessed by colony-forming unit-granulocyte and macrophage ( CFU-GM) assay. Results Fifteen days after busulfan treatment, weight, white blood cell (WBC), red blood cell (RBC), blood platelet (PLT), lymphocyte (LYM), and neutrophil (NEU), as well as percentages of HPC, HSC, and LT-HSC in the bone marrow of KKA^y mice were significantly reduced. In addition, the number of CFU-GM was significantly reduced, although proportions of spleen and thymus cells were not significantly changed. After busulfan treatment, WBC, RBC, HGB, and LYM of KKA^y mice were significantly reduced compared with B6 mice, but proportions of PLT and HSC were significantly increased. PLT and HSC were significantly reduced in KKA^y mice compared with B6 mice, while the decrease of HPC in KKA^y mice was significantly enhanced compared with B6 mice. Conclusions Busulfan (40 mg / kg) impaired function of the hematopoietic system in mice. After busulfan was administered, WBC, RBC, HGB, and LYM of KKAy mice were severely damaged, PLT and HSC were less damaged and more tolerant, and the tolerance of HPC was reduced compared with B6 mice.

Abstract:Objective To establish a rapid and efficient method for determining right ventricular pressure in mice without the aid of a ventilator and explore the measurement techniques. Methods Twenty-three C57BL/ 6 J mice (8 weeks old) were randomly divided into two groups: control group ( n = 12) and hypoxic group ( n = 11). The hypoxic group was reared in a hypoxic chamber (10% O2 _{) for 4 weeks to prepare the pulmonary hypertension model, while the control group was reared in normal air; otherwise, the conditions were the same. Then, we connected a multi-channel physiological instrument and measured right ventricular pressure in mice with a 0. 6×25 mm disposable infusion needle by puncturing the intercostal muscle directly when the chest cavity was closed. Thoracotomy was performed to verify the puncture point. Next, the heart was separated, the right ventricle (RV), and left ventricular and septum (LV + S) were weighed, and the right ventricular hypertrophy index ( RVHI) was calculated. Pathological sections of left lung were taken and stained with hematoxylin-eosin and α-smooth muscle actin to observe the structural changes of pulmonary artery. Results A right ventricular pressure curve was obtained by directly puncturing the right ventricle through the intercostal muscle, and the whole process was completed within 3- 4 minutes; the success rate of RVSP measurement was 95. 6% ( 1 failed in the control group). The average RVSP and RV/ (LV + S) of the hypoxia group were 30. 48±2. 95 mmHg and 35. 31±3.78%, which were higher than those of the control group with 17. 50±3. 40 mmHg and 23. 14±4. 29% ( both P < 0. 05), respectively. In the hypoxic group, the pulmonary arterioles were thickened and the smooth muscle of the tunica media proliferated. Conclusions The method of measuring the right ventricular pressure curve of mice by intercostal muscle puncture and keeping the animal chest closed without ventilator-assisted ventilation is simple and fast; it can rapidly assess the hemodynamic state of the right ventricle of mice with a high success rate and is worthy of being popularized.}

Abstract:Objective To study the age-related changes in SVZ gene expression of 3-week-old, 6-month-old, and 20-month-old mice. Methods RNA was extracted from the SVZ of mice at the three ages. The difference in gene expression of SVZ at the different ages was detected by RNA sequencing and quantitative real-time PCR. Results Compared with the levels in young mice, 253 differentially expressed genes were detected in adult mice and 519 in old mice. Compared with the levels in adult mice, 147 differentially expressed genes were detected in old mice. GO analysis found that the differentially expressed genes are mainly involved in nutrition and metabolism in adult mice compared with young, and central nervous system neuron differentiation among others in old mice compared with adult mice. KEGG analysis found that the differentially expressed genes are mainly involved in protein digestion and absorption, and neuroactive ligand–receptor interaction, among others. The differentially expressed genes were analyzed by GO and KEGG pathways analyses. Conclusions This study provides basic data on the changes of gene transcription levels with age in the SVZ, and also provides a new theoretical basis for treatment and resistance to nerve aging in the future.

Abstract:Objective To isolate, culture, identify, and analyze the drug sensitivity of pathogenic bacteria in inflamed lung tissue from a dead rhesus monkey, to determine the drug susceptibility of the bacteria, and to guide proper clinical application of such information. Methods The rhesus monkey was dissected under aseptic conditions. The lung was cut open and then insert focus of infection in the lung with inoculating loop to sampling and inoculating on plating medium. The presence of bacteria was observed, biochemical identification was performed, and eight kinds of antibiotic susceptibility test were carried out. Results Bacterial colonies on SS agar medium were white or ivory white, round,moist, and opaque. After the biochemical identification, the bacterium was identified as Enterobacter cloacae (ID: 99. 9%, T: 0. 71, ID number: 35074753311). The drug sensitivity test showed that the susceptibility of Enterobacter cloacae to eight antimicrobial agents was as follows: ampicillin and cefuroxime, intermediate sensitivity; and norfloxacin, amikacin, gentamicin, cefatriaxone, cefoperazone, and cefotaxime, saitivity. The Enterobacter cloacae isolated from rhesus monkey also showed a high risk to Kunming mice. Conclusions We conclude that the rhesus monkey lung was infected with Enterobacter cloacae. Cefoperazone and cefotaxime can be used clinically to treat cases like this in rhesus monkey.

Abstract:Objective To provide approaches for modeling the development of anti-hyperuricemia drugs, acute and chronic hyperuricemia animal models were established, and positive drug was applied to verify the curability of these two animal models. Methods The acute hyperuricemia mouse model was established by the intraperitoneal injection of hypoxanthine and subcutaneous injection of oteracil potassium. Allopurinol tablets were administered to the positive group half an hour after the model establishment. Finally, the concentrations of serum uric acid, serum creatinine, and serum urea nitrogen and the activity of liver xanthine oxidase (XOD) were determined 2 h after model establishment. In contrast, adenine and ethambutol dihydrochloride were used to establish the chronic hyperuricemia rat model by gavage. The positive group was administered allopurinol tablets daily. After 21 days, the concentrations of serum uric acid, serum creatinine, and serum urea nitrogen, the activity of XOD, and the histopathological change of kidneys were determined. Results The concentrations of serum uric acid and serum creatinine were significantly elevated (P< 0. 05) in both the acute and the chronic hyperuricemia models. Additionally, serum urea nitrogen and XOD activity were also clearly elevated (P<0. 05), while renal tubule interstitial injury and urate crystallization could be found, the scores of which were significantly increased in the chronic hyperuricemia model. However, the concentration of serum uric acid was decreased after treatment with allopurinol tablets for both acute and chronic hyperuricemia animals; moreover, in the chronic hyperuricemia animals, the concentration of serum creatinine and activity of liver XOD were decreased, while kidney injury and urate crystallization were also improved. Conclusions The approaches established in this study are suitable for creating acute and chronic hyperuricemia animal models, and can be used for drug pharmacodynamic observation.

Abstract:Objective To identify the genetic pattern and locate the genes related to late-onset congenital cataract FVB mouse. Methods First, the phenotype of cataract mice was identified using histopathological slides and the genetic pattern of congenital cataract phenotype was identified by constructing pedigrees. Second, the linked chromosome was located by whole-genome SNP scanning based on multiplex PCR targeted sequencing. Finally, screening of candidate mutant genes was performed by sequencing the whole exome of two cataract mice, one wild-type mouse, and four F2 cataract mice with a mixed genome. Results The result of whole-genome scanning of 100 F2 mice showed the highest linkage between rs4228772 SNP on chromosome 11 and the cataract phenotype, with a 82. 11% homozygous rate. The result of whole-exome sequencing further demonstrated three genes that had spontaneously mutated on chromosome 11:Sfi1,Obscn,andPtrh2. Conclusions The combination of whole-genome SNP scanning and whole-exome sequencing strategies revealed that congenital late-onset cataract was caused by mutated genes on chromosome 11 and inherited as a dominant trait. This method may also be applied to the study of gene function in other model mammals with a clear genetic background.

Abstract:Objective To investigate the effects of SOX6 gene interference on apoptosis, mitochondrial membrane potential, and the PI3K/ AKT signaling pathway of astrocytes (AS) induced by H₂O₂ . Methods AS cells were isolated from the cerebral cortex of neonatal rats and divided into the following groups: negative control group (NC group): AS transfected with siRNA without interference; H₂O₂ group: AS cells treated with 20 mol/LH₂O₂ for 24 h; H₂O₂ +NC group: non-interfering siRNA transfected into AS cells for 24 h, then treatment of cells with 20μmol /L H₂O₂ for 24 h;and H₂O₂+si-SOX6 group: SOX6-specific siRNA transfected into AS cells for 24 h, followed by treatment of the cells with 20 μmol / L H₂O₂ for 24 h. The cytotoxicity, apoptosis rate, and mitochondrial membrane potential were determined by LDH kit, flow cytometry, and JC-1 probe. The expression of SOX6, Cyt. C, caspase 3, caspase9, Bcl-2, Bax, and p-AKT protein was determined by Western blotting. Results H₂O₂ significantly upregulated the expression of SOX6, Cyt. C, caspase3, caspase9, and Bax; downregulated the expression of Bcl-2 and p-AKT; increased cytotoxicity; and induced apoptosis. Interfering with SOX6 gene expression significantly reduced the effects of H₂O₂ on SOX6, Cyt.C, caspase3, caspase9, Bcl-2, Bax, and p-AKT expression, cytotoxicity, and apoptosis. Conclusions Interference with SOX6 gene expression can protect against AS injury induced by H₂O₂ through the anti-apoptotic pathway, which is related to the mitochondrial pathway and PI3K/ AKT signaling activation.