Abstract:Objective To establish reference values for blood pressure in cynomolgus monkeys in different ages. Methods The blood pressures and blood lipids indexes were detected in 521 cynomolgus monkeys using an American BECKMAN-CX4 automatic biochemical analyzer and a wrist electronic blood pressure monitor. Statistical tests were performed to analyze the data. Results Significant differences were found in blood pressure values of cynomolgus monkeys in different ages. Blood pressure values in the elderly group were higher than those of other groups. The morbidity of hypertension in the elderly group was higher than those of the other groups. Body mass index (BMI) in the hypertension group was higher than that of normal group in the same age. The incidence of hypertension in the elderly group with hyperlipemia was higher than that of other groups. Logistic regression analysis showed that age, BMI and hyperlipidemia in the hypertensive group were 1.435, 1.218, and 2.337 times higher than those of the normal group when predicting the risk of hypertension. Conclusions We have initially established reference values of blood pressure in cynomolgus monkeys in different ages. Age, BMI and hyperlipidemia are risk factors of spontaneous hypertension in cynomolgus monkeys, and the measurement of blood pressure may provide a basis for the screening of cynomolgus monkey model of spontaneous hypertensive and related research.

Abstract:Objective To analyze the effect of intrathecal injection of dexmedetomidine on the behavioral activity, pain degree and expression of protein kinase C in spinal dorsal horn of rat models of chronic neuropathic pain, and to investigate the analgesic mechanism of dexmedetomidine. Methods 75 healthy male rats were randomly divided into observation group, model group and control group, 25 rats in each group. Chronic sciatic nerve injury model was established in the observation group and model group. After modeling, intrathecal dexmedetomidine intervention was used in the observation group. The model group was treated with saline injection and there was no intervention in the control group. Before the modeling (BM)and at 3(D3), 5(D5), 7(D7), and 14 (D14)days after medicine administration, the behavioral capacity was evaluated by cumulative evaluation method and movement function evaluation, and the assessment of pain degree (mechanical withdrawal method and thermal withdrawal latency pain threshold detection method), PKC staining score (immunohistochemical SABC method), PKC mRNA assay (RT-PCR method) and PKC protein expression (Western blot) were conducted and the data were statistically analyzed. Results ① Before modeling, the behavior, the cumulative scores of motor function, MWT, and TWL showed no significant differences between the different groups (P>0.05). After modeling, the model group and observation group showed that the cumulative scores and motor function scores were increased significantly, MWT and TWL decreased significantly, and the changes in the observation group were significantly lower than those in the model group (P<0.05). After modeling, the cumulative scores, motor function scores, MWT, and TWL were significantly different between the groups (P<0.05). ② The expression of PKC was negative in the control group and positive in the model group. In the observation group, after the initial establishment of model, the PKC was strongly positive, and along with the prolonged treatment, the PKC expression intensity was decreased, and only weakly positively expressed at 14 d. ③ After modeling, the observation group and model group showed that the PKC mRNA and PKC protein expression levels were significantly higher than that of the control group (P<0.05). With the continuous drug administration, the PKC mRNA and PKC in the observation group were decreasing, and reached a level close to that of the control group at 14 d of drug administration. From the third day after modeling, at the same time points, the amount of PKC expression in the observation group was significantly lower than that in the model group (P<0.05). Conclusions Intrathecal injection of dexmedetomidine can improve the behavior of rat models with chronic neuropathic pain, and reduce the degree of pain. It may be related to the inhibition of protein kinase C expression in the spinal dorsal horn.

Abstract:Objective To observe the sensitivity of Wuzhishan, Tibetan and Bama minipigs to exogenous fats. Methods A total of 15 male minipigs including 5 WZS minipigs, 5 Tibetan minipigs and 5 Bama minipigs, were used in this study. The minipigs were intravenously injected with fat emulsion and fed with high-fat diet, and the changes of serum total cholesterol (TC), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C) and triglyceride (TG) levels were detected at 0.5 h, 1 h, 1.5 h, 2 h, 2.5 h and 3 h after intravenous injection of fat emulsion and at 3 h and 12 h, 24 h, 36 h, 48 h, 60 h, 72 h and 84 h after fed with high-fat diet, respectively, and calculated the changes of area under the curve (ΔAUC) values. Results The triglyceride (TG) in the three kinds minipigs were significantly increased (P<0.01), and reached a peak at 0.5 h after injection. The degree of elevation of ΔAUC values were different showing on increasing order: Tibetan minipig > Wuzhishan minipig > Bama minipig, while TC, LDL-C and HDL-C showed no significant changes (P>0.05). Moreover, TC, LDL-C, HDL-C and TG were significantly increased in the three stocks of minipigs induced by feeding with high-fat diet (P<0.05, P<0.01). Among them, the TC and LDL-C of Wuzhishan minipigs peaked at 36 h, HDL-C peaked at 48 h and TG peaked at 24 h after feeding, respectively. TC, LDL-C and HDL-C in the Bama minipigs and Tibetan minipigs peaked at 48-60 h and TG peaked at 36 h after feeding, and the ΔAUC values of TC, LDL-C and HDL-C were in an increasing order of Wuzhishan minipigs > Bama minipigs > Tibetan minipigs. Conclusions The three stocks of minipigs are sensitive to TG after intravenous injection of fat emulsion, and the lipid tolerance values are in an order of Tibetan minipig > Wuzhishan minipigs > Bama minipigs. Meanwhile, the three stocks of minipigs are also sensitive to TC, LDL-C and HDL-C after feeding with high-fat diet, and the lipid tolerance values are in an increasing order of Wuzhishan minipig > Bama minipigs > Tibetan minipigs.

Abstract:Objective To investigate the changes of the platelet function in APP/PS-1/tau(3xTg) mice, a murine model for Alzheimer's disease,and explore its mechanisms. Methods We assessed the change of function of platelet in 3xTg-AD mice by flow cytometry. Adhesion assay and Western blotting were used to compare with the data of wild type mice. Results Platelets from aged 3xTg-AD mice were normal in number and glycoprotein expression (P> 0.05), but adhere more avidly on matrices such as fibrinogen, compared with the platelets from age-matching wild type mice (P<0.05). The washed platelets of 3xTg-AD mice were adherent to fibrinogen, and also showed increased phosphorylation of selected signaling proteins, including PI3 kinase effector Akt and p38MAP kinase (P<0.05). In contrast, activation induced by several agonists in 3xTg-AD mice was similar to that of wild type platelets (P>0.05). Conclusions These results demonstrate that Alzheimer's mutations result in a significant hyper-activated adhesion state of circulating platelets, evident with the progression of the disease.

Abstract:Objective To study the effect of heterozygous deficiency of SGT gene on the hematological and biochemical parameters of mice in young and elderly ages.Methods Blood samples were analyzed for complete hematological and biochemical parameters from heterozygous SGT-deficient and wild-type mice of 10-weeks and 6-months old mice, respectively. Results Age-related changes in most indexes were found statistically significantly different between young and elderly mice with the same genotype. Compared with the wild type at the same age, the platelet large cell ratio (P-LCR) was lower in young heterozygous SGT-deficient mice. However, platelet count, plateletcrit (PCT) and neutrophil count were more significantly lower in elderly heterozygous SGT-deficient mice (P<0.05). There was no significant difference for biochemical parameters ALT, AST, LDH, urea nitrogen, creatinine and blood glucose. Total and unconjugated bilirubin as well as ALP were significantly higher in elderly heterozygous SGT-deficient mice but not for conjugated bilirubin (P<0.05). In addition, significant differences existed for the lipids between two elderly groups (P<0.05). Conclusions Heterozygous deficiency of SGT gene induced changes of some hematological and biochemical parameters in elderly mice. It provides helpful information for further investigation on SGT involvement in some biological and pathological processes.

Abstract:Objective To explore the effect of inhibition of miR-214 expression on the proliferation of hepatocellular carcinoma cells via regulation of E2F3 expression. Methods The expression of miR-214 in SMMC-7721, HepG2, SK-Hep-1 and Huh 7 cells was examined by RT-PCR. Hepatocellular carcinoma cells were transfected with miR-214 NC and miR-214 mimics with liposomes. The expression of miR-214 was detected by RT-PCR. The cell viability was detected by MTT assay. Cell apoptosis was detected by Hoechst staining. Cell cycle was detected by flow cytometry. Western blot, RT-PCR and dual luciferase reporter gene assay were used to detect whether E2F3 was a downstream target gene of miR-214. Results The expression of miR-214 in SMMC-7721, HepG2, SK-Hep-1 and Huh 7 cells was 0.83±0.08, 0.32±0.03, 0.33±0.03, and 0.08±0.01, respectively. The expression of miR-214 in the HepG2 cells was the lowest, so HepG2 cells were selected as the subsequent experimental cell line. Compared with the miR-214 NC group, the expression of miR-214 (0.65±0.06 vs. 0.14±0.01) was up-regulated, the cell viability (0.35±0.03 vs. 0.69±0.06) was decreased, cell apoptosis rate [(36.37±3.43)% vs. (3.74±0.34)%] was increased, the G1 phase of the cell cycle (57.79±5.78 vs. 45.319±4.53) was prolonged, the expression of E2F3 protein (0.23±0.02 vs. 0.98±0.09) and mRNA (0.24±0.02 vs. 0.99±0.10) was significantly down-regulated in the miR-214 mimics group (P<0.01). Conclusion miR-214 mimics suppress the HepG2 cell proliferation via targeted down-regulation of E2F3 expression.

Abstract:Objective To establish a C57BL/6 mouse model of intestinal infection induced by S. Typhimurium. Methods In order to improve the infectious sensitivity of S. Typhimurium, C57BL/6 mice were intragastrically given 5% (w/v) NaHCO_3. Then mice were challenged with S. Typhimurium. The health condition, survival and body weight of mice were observed from day 0 to day 7 after the bacterial infection. The pathological changes were also examined. Results the mice challenged with S. Typhimurium showed decreased body weight and typical clinical signs, including in appetence, piloerection and low survival rate. Macroscopic dissection revealed that intestinal hyperemia and swelling were founded in the mice challenged with S. Typhimurium. Histopathology showed intestinal epithelial and mucosal damages. Conclusions We have successfully established a C57BL/6 mouse model of S. Typhimurium infection. This model may be of crucial significance for studying the biological functions of associated immunological molecules or cytokines in the process of inflammatory bowel disease induced by S. Typhimurium.

Abstract:Objective To report a newly developed method and procedure to establish a rat model of subarachnoid hemorrhage in detail, and to provide a better model simulating the clinical subarachnoid hemorrhage caused by a ruptured aneurysm for related research. Methods One hundred and twenty healthy SPF 2-3-month old male Sprague-Dawley rats were divided into 4 groups, 30 rats in each group. The three experimental groups were sacrificed at 6, 24 and 72 hours after modeling. Rat models of subarachnoid hemorrhage were established by inserting a fiber core in the internal carotid artery and piercing this artery. Successful establishment of the subarachnoid hemorrhage model was confirmed by observation of breathing, pupil, defecation, urination and inspection at autopsy dissection. The controllability and reproducibility of this model were verified by observation of clinical manifestation and explored by mortality analysis. Results Subarachnoid hemorrhage was successfully induced by fiber core piercing the internal carotid artery at the needed location. Conclusions This method of model preparation is stable and understandable. The operation is nimble, with a good reproducibility. This model can be successfully performed after a short time learning, well simulate the sudden hemorrhage caused by a ruptured aneurysm, and suitable for research on early brain injury and vasospasm after subarachnoid hemorrhage.

Abstract:Objectives To observe the anesthetic effects of an improved anesthetic method on goat by comparing with the original method, and to provide theoretical and practical reference for anesthesia during the establishment of goat models. Methods A total of 72 goats were selected to make tibial fracture models, 36 were enrolled as group A and others as group B. The group A was injected with Sumianxin and diazepam. The group B was injected with Sumianxin, diazepam and pumped propofol by intravenous pump. The times of animal struggle, total dose of additional Sumianxin, death rate, and recovery time were evaluated. Results The death rate of group A was 16.67% and there was no death in the group B. The times of struggle, the total dose of additional Sumianxin and the recovery time of the group A were higher than group B, showing a significant difference (P<0.05). Conclusions The improved anesthetic method can decrease the times of goat struggle times, reduce the dose of anaesthetics, and lower the death rate.

Abstract:Objective To observe the morphological features of mouse cornea after fixation with 6 different types of fixatives, and choose an optimal fixative suitable for fixation of mouse cornea. Methods 60 eyes of NIH mice were randomly divided into 6 groups, 10 eyes in each group. The eyes were respectively fixed in the 6 types of fixatives, namely, 95% ethanol, 10% neutral buffered formalin, 10% formalin, Bouin's solution, Formaldehyde-acetic acid fixative solution, and Davidson's solution. After fixation, the specimens were dehydrated with 80%, 95%, 100% gradient alcohol, and clearing was performed using TO-type histological clearing agent, then soaked in melted wax, and finally embedded in paraffin. The paraffin sections were 4 μm thick, and stained with hematoxyline and eosin. The histological features of the cornea were observed and various regions of the cornea were measured. Results Different fixatives showed different histological characteristics and thickness of the mouse cornea differently. The acetic acid-formaldehyde fixative caused least changes of the morphology and thickness of the mouse cornea.Conclusions It is important to consider the impact of different fixatives on the corneal tissue morphology. In order to avoid histological artifacts, formaldehyde-acetic acid fixative among the 6 examined fixatives is the most suitable for histological examination of mouse cornea.

Abstract:Objective To provide a simple, convenient, and safe anesthesia method for the establishment of a M1 segment of middle cerebral artery occlusion model in rhesus monkey or other large laboratory animals. Method Twenty male rhesus monkeys weighing 7-11 kg (ages 7-9 years) from Academy of Military Medical Sciences were used in this study. Sumianxin injection combined with 0.1 mg/kg ketamine was given before endotracheal intubation (ID:4.5-5.5#). The animals were then transported to an interventional operation room, where the intravenous access was established and a urinary catheter was inserted into the urinary bladder. Mechanical ventilation was used during the surgery, propofol was continuously injected in a speed of 2-4 mg/kg/h, and Sumianxin-ketamine could be given if necessary to maintain adequate anesthesia depth. The dose was adjusted according to vital signs of the rhesus such as body movements, physiological parameters, and demand of surgery. Brain MRI examination was performed before and after thrombolysis. Anesthetic injection was suspended and the animals were allowed to have a spontaneous breathing every time before the MRI text. Heart rates, temperature, non-invasive blood pressure, and SpO₂ were monitored during the whole surgery. Blood samples were taken from the radial artery for blood gas analysis after anesthesia induction and during operation. Results All the 20 animals underwent the operation successfully, no animal had restlessness, respiratory depression, arrhythmia and other serious complications. At the end of the study, animals awake soon. Fifteen of them survived longer than 24 hours, only 5 died from serious cerebral hemorrhage and larger cerebral infarction. Conclusions General endotracheal anesthesia is safe for rhesus monkeys during such interventional operation and MRI examination.

Abstract:Objective To establish a rapid, simple, sensitive, and specific multiplex real-time PCR method for quantitative detection of Campylobacter jejuni, Salmonella and Shigella in tree shrews. Methods Specific primers and probes were designed, according to the HipO gene of Campylobacter jejuni, inV gene of Salmonella and ipaH gene of Shigella. The primers were confirmed by single pathogen quantitative PCR, and the sensitivity and specificity of the multiplex PCR were analyzed. Finall, the samples of experimental tree shrews were detected by this multiplex PCR method. Results The PCR element of TaqMan-MGB real-time PCR assay was able to quantitatively amplify the Campylobacter jejuni, Salmonella or Shigella. Appropriate standard amplification curves of Campylobacter jejuni, Salmonella and Shigella in the multiplex quantitative PCR were obtained. The sensitivity of this method was 1×10³ ng/μL. There was no false positive detection from other bacterial strains. Conclusions This multiplex quantitative real-time PCR method has good application and development prospects in the detection of microorganisms in tree shrews.

Abstract:The quarantine inspection and acceptance of laboratory animals is an important work, which can protect animals from pollution, occurrence and spread of diseases in the surounding area, and it is the key point to realize the quality standardization of laboratory animals. Beagle dogs are acknowledged widely as specialized laboratory dogs which is widely employed in experiments of drug safety evaluation because of the good genetic stability, environmental adaptability, disease resistance and consistency testing in the experiments. Establishment of standard operating procedures of beagle dog quarantine acceptance check for drug GLP organization tests, refining technical points, strengthen the technical training of quarantine officers, and efforts to improve the level of quarantine are needed to finally ensure the quality of laboratory animals.

Abstract:Animal experiment on influenza virus infection carries certain biohazard risk, with a threat to the health of researchers and public health. The risk levels differ by influenza virus types and subtypes. This article combs the domestic and national laws and rules, and explores the biosafety management of animal study on influenza virus.

Abstract:There are approximately 240 million patients with chronic hepatitis B virus (HBV) in the world. Despite the production of HBV vaccines and antiviral drugs, HBV still remains a serious threat as an infectious disease to human health. Due to the lack of an ideal animal model, the study of HBV is limited. This review was to analyze in accordance with existing HBV animal model, to explore the differences in applications of various models regarding their etiology, pathology and pathophysiology. thus, to provide a reference for researchers in future HBV research to better use the resources.

Abstract:Plasmacytoid dendritic cells (pDCs) as innate immune cells can produce a large amount of interferon-alpha (IFN-α). During the stage of acute human immunodeficiency virus (HIV) infection, pDCs can inhibit HIV replication by releasing IFN-α and activating adaptive immune responses. In the stage of chronic HIV infection, pDCs play a role in immune suppression by regulating immunocytes and damage of the immune system by depletion of the lymphocytes. Finally, pDCs have influence on the disease progression of acquired immune deficiency syndrome (AIDS).

Abstract:CD45 is a transmembrane molecule with phosphatase activity, and plays a major role in immune cells. CD45 is required for the antigen receptor signal transduction, and attributed as an apoptosis regulator. Impairment of this function may result in autoimmune, immunodeficiency, and malignant diseases. The role of CD45 in HIV-1 infection is one of important research topics. This paper summarizes the research progress on the role of CD45 in HIV-1 infection.

Abstract:Zebrafish (Danio rerio), as a new vertebrate model organism, has been widely used in human disease research in recent years, especially in the study of human bone diseases. The zebrafish models show some advantages, such as expression of disease-related genes and application in pharmacodynamic screening and evaluation, etc. It is also of great practical significance for the traditional medicine, which has a characteristic efficacy in the treatment of bone diseases. This paper reviews the research and application of zebrafish in research on bone diseases such as osteoporosis, fracture, osteoarthritis, and so on, aiming to provide some inspiration for the future clinically relevant research.

Abstract:Right ventricular failure(RVF)is a severe clinical syndrome that affects human health seriously.The establishment and application of experimental animal models are critical to research the pathogenesis, pathophysiology, prevention and treatment of RVF. Recently, numerous researchers have studied the modeling methods and testing indexes of RVF of animal models, which has improved the success rate and quality of the models. However, even if so many models, there is still no widely accepted experimental animal model of RVF because of the different and complex modeling methods, material selection and experimental conditions. This article reviews the methods and mechanism of multiple experimental animal models of RVF and compares them, so as to provide reference for choosing and applying experimental animal models of RVF in the study.