Abstract:Objective To study the effects of morroniside on the expression of the tissue inhibitor of metalloproteinase (TIMP) of the ischemic ipsilateral cortex 7 days after ischemia reperfusion. Methods 25 male Sprague-Dawley rats were subjected to middle cerebral artery occlusion (MCAO) model with modified Zea Longa's method, and then divided them into sham group (n=5), ischmia group (n=5), and morroniside groups (low, medium, and high dosage groups, n=5) randomly. Morroniside were then administered intragastrically once a day at dose of 30 mg/kg, 90 mg/kg and 270 mg/kg 3 hours after operation. The expression of TIMP of the ischemic ipsilateral cortex on 7 days after ischemia-reperfusion were detected by western blotting and immunofluorescent staining analysis. Results Compared with the sham group, the expression of TIMP in ischemia group increased (P < 0.05) 7 days after MCAO. Compared with the ischemia group, after treatment with morroniside at doses of 90 mg/kg and 270 mg/kg, the expression of TIMP increased significantly (P < 0.05; P < 0.01). Conclusions Morroniside could increase the expression of TIMP in the ischemic ipsilateral cortex and may contribute to blood-brain barrier protection and restoration.

Abstract:Objective To investigate the effect of SWF on the behaviors and striatal levels of amino acids neurotransmitters of LID model rats,and to explore the effect of SWF on the mechanism of dyskinesias. Methods SD rats were injected with 6-hydroxydopamine (6-OHDA) to produce hemi-lateral nitro-striatum lesion. 6-OHDA lesioned rats were daily given L-DOPA, or L-DOPA plus SWF at different doses. Sham group was also established. All rats were treated for 22 days, once per day. During the treatment, abnormal involuntary movements (AIMs) were evaluated. On 22nd day, in vivo microdialysis was used to collect striatal extracellular fluid as the rats were in sober and free-moving condition. All dialysates were analyzed by high performance liquid-fluorescence (HPLC-FLD) detection. Results After treatment of L-DOPA, 6-OHDA lesioned rats developed AIM. AIM scores of L-DOPA-treated rats were higher than the sham-operated rats (P < 0.01). L-DOPA plus high dose of SWF-treated rats had lower AIM scores than L-DOPA-treated rats (P < 0.05). On 22nd day, in L-DOPA-treated rats, striatal levels of glutamate and GABA increased (P<0.05) than sham group. 60 min after last treatment, levels of glutamate and GABA in L-DOPA-plus SWF-treated rats were significantly lower than the sham-operated and L-DOPA plus SWF-treated rats (P < 0.05, P < 0.01). Conclusions The findings in this study suggested that as SWF was combined, side effects of L-DOPA mitigated as AIMs scale rating were reduced in LID rats. SWF protected 6-OHDA lesioned rats from LID by regulating abnormal striatal concentrations of glutamate and GABA.

Abstract:Objective To investigate the effect of chronic corticosterone administration on depressive-like behaviour and synapsin-1 expression in mice. Methods C57BL/6J mice were randomly separated into 2 groups (n=10 per group): control and Corticosterone (CORT). CORT was dissolved in vehicle (0.45%hydroxypropy-β-cyclodextrin, β-CD). Corticosterone (5mg/kg/day) was delivered for 35 days in drinking water and continued when the behavioral tests were performed. Control animals received vehicle(β-CD) in drinking water during the entire experiment. The behavior tests were applied to evaluate depressive-like behavior of the model. Results The sucrose preference test showed no significant difference in sucrose intake between control and CORT group (P > 0.05). The immobility time was increased in the forced swim test (P < 0.05). The cumulative duration,distance moved and frequency in the center area were decreased in the open field test(P < 0.05,P < 0.01,P < 0.01). The cumulative duration in the border area was increased(P < 0.01). Latency to food was decreased in the novelty suppressed test(P < 0.05). Immunohistochemisty analysis showed a notably decreased expression of Syn-1 in the hippocampus CA1,CA3 and DG. Conclusions Chronic administration of CORT induced depressive-like behavior and decreased Syn-1 expression.

Abstract:Objective To investigate the effect of Low molecular weight heparin sodium (LMWH-Na) adjuvant on humoral immune response induced by hepatitis A virus(HAV)vaccine in mice. Methods ICR mice were randomly divided into 8 groups, six for each, and the mouse of each group are injected respectively with live attenuated hepatitis A vaccine HepA-1(18 EU) +different doses (100 μL, 20 μL, 10 μL, 1 μL, 0.1 μL) of LMWH-Na, while those injected with physiological saline as a negative control group (Blank), those injected with HepA- 1(18 EU) as positive control group, those injected with Al(OH)₃(300 μg)+HepA-1(18 EU) as the aluminium adjuvant control group, 200μL for each. The specific IgG level against HAV in sera of mice at weeks 4, 8, 12 and 16 after immunization were determined by indirect ELISA. During the experiment, the health status of the mice were observed, the kidney, spleen, liver, lung and heart of mice immunized with LMWH-Na 100 μL at the maximum dosage and those in negative control group were taken out at week 16 after immunization, and prepared into sections for pathological observation. Results In addition to the negative control group, mice in various group produced anti -HAV IgG at 4 weeks after the immunization, of which the levels increased as time went on, and reached peak values at week 8 then decreased gradually. At 4 week and 8 week, the anti -HAV IgG levels of aluminum adjuvant control group, LMWH-Na 100 μL group, LMWH-Na 20μL group, LMWH-Na 10 μL group were higher than the positive control group and the difference was statistically significant (P < 0.05). At 12 week, the anti -HAV IgG levels of aluminum adjuvant control group, LMWH-Na 20 μL group and LMWH-Na 10 μL group were higher than the positive control group and the difference was statistically significant (P < 0.05). 16 week,the anti -HAV IgG levels of aluminum adjuvant control group and LMWH-Na 10 μL group were higher than positive control group and the difference was statistically significant (P < 0.05),but the anti-HAV IgG levels of various LMWH-Na groups were lower than the aluminum adjuvant control group. Conclusions LMWH-Na can obviously enhance the HAV antigen induced humoral immune response in mice,and have a value of becoming a new type of vaccine adjuvant.

Abstract:Objective To explore the modified increasing intracranial pressure method to set up the brain death model for rabbits and sustain the state of brain death. Methods 15 healthy New Zealand rabbits were divided into brain death group (Group A,n=9) and sham operation group(Group B,n=6) randomly. The 9 brain death rabbits were set up by increasing intracranial pressure in a modified way. The sham operation rabbits were only sustained with anesthesia.Recorded the mean artery pressure (MAP) and heart rate (HR). Results In group A, the 8 brain death rabbits were successfully set up. Only 1 died of heart failure. Compared to Group B, the MAP and HR of Group A rise up greatly during increasing intracranial pressure and presented an increasing trend. The peak MAP and HR were (111.63±7.71) mmHg and (262.38±16.60)beats/min which were much higher than the MAP(80.63±8.45) mmHg and HR(153.25±14.35) beats/min before increasing intracranial pressure(P < 0.05). After brain death, MAP decreased sharply to 45mmHg. 2 hours after brain death, HR (218.50±10.27) beats/min presented an decreasing trend too, which showed a significantly statistic difference to peak HR(262.38±16.60) and HR(153.25±14.35) beats/min before brain death(P < 0.05). Conclusions Compared to traditional method, the brain death model could be established stably and securely in the modified way and maintained for long time with the respiration and circulation supports.

Abstract:Objective To preliminary study nutrition requirement of Mugilogobius chulae in the transitional period and provide reference data for the research and development of compound feed and relevant standards. Methods Artemia (Artemia nauplii, CK) and 7 commercial compound feeds (P30L8, P35L8, P50L8, P55L8; P50L12, P53L12, P55L12) were used to feed goby in transitional period. At the end of 8 weeks growing experiment, some data like survival rate (SR), weight gain (WG) and specific growth rate (SGR) and so on were measured. While, 8 groups' body compositions were also measured. Results SR Treatment groups (51.78±8.98～68.22±4.95) and CK (58.22±6.51) have no significant different (P > 0.05). WG Treatment groups except P30L8 (130.76±23.67) and P50L12 (220.86±16.9) were significant larger than CK (206±41.91, P < 0.05). Using equal lipid (8% and 12%) dietary, WG increased with protein level rising.SGR Treatment groups' SGR also increased with protein levels rised.FR, FCR P50L8, P55L8; P50L12, P53L12, P55L12's FR and FCR were better than group CK (P < 0.05). Using equal lipid (8%) dietary, FR and FCR were decreased with protein level rised. In addition, different feed feeding group's body composition showed no significant difference (P < 0.05). Conclusions Rear goby with compound feed replace the living food was viable and supported appreciable growth. Their crude protein and lipid have no effect to the goby's body composition (P < 0.05). Dietary with crude protein 50% and crude lipid 8% could obtain a better growth in transitional period.

Abstract:Objective To illuminate energy demands of Mongolian gerbil by analyze the diversity in intestinal morphology and the energy metabolism of different species. Methods 10 SD rats, 10 ICR mice and 10 mongolian gerbils were feed individually in metabolic cages. Gather and measure the energy of their feces and urine in 24 h for 5days. Then, all the animals were weighed and anaesthetized, collect and measure the length of the intestine. Results Rats digestive energy and metabolic energy respectively are 314.56 kJ/d and 314.55 kJ/d, mice are 10.608 kJ/d and 9.799 kJ/d, gerbils are 99.828 kJ/d and 99.927 kJ/d. Rats digestive energy and metabolic energy used to increase body weight respectively are 19.273 kJ/g and 18.831 kJ/g, mice are 71.842 kJ/g and 72.390 kJ/g, gerbils are 56.142 kJ/g and 55.965 kJ/g. In these animals, the length of gerbils' digestive tract is the shortest, the length of rats' small intestine is the longest, and the percentage of gerbils' appendix is the largest. Conclusions There are significant differences in the length of the gut and energy needs.

Abstract:Objective To investigate the influence of acupuncture,warm-needle acupuncture, electro-acupuncture on cartilage metabolism-related factors of the experimental knee osteoarthritis model, and explore the similarities and differences of mechanism in different kinds of acupuncture treatment. Methods New Zealand rabbits were equally randomized into control, model,acupuncture, warm-needle acupuncture and electro-acupuncture groups. KOA model was duplicated by anterior cruciate ligament transaction (ACLT). After KOA model was established one week, acupointsof“Dubi”(ST35),“Zusanli”(ST36) were used for treatment with different acupuncture treatmentstwenty times totally. The concentration of IL-1β, TNF-α, TGF-β1,MMP-1,MMP-3 and TIMP-1 in serum were detected by enzyme linked immunosorbent assay(ELISA). Results In comparison with control group, the concentration of IL-1β, TNF-α, TGF-β1,MMP-1 increased significantly in model group(P < 0.05),while MMP-3 reduced significantly(P < 0.05); compared with model group, IL-1β,TNF-α, TGF-β1,MMP-1 reduced significantly in each treatmentgroup(P < 0.05 or P < 0.01),MMP-3 increased significantlyin warm-needle acupuncture group(P < 0.05); comparison of IL-1β,TNF-α, TGF-β1,MMP-1in serum was not statistically significant (P >0.05) between treatment groups; MMP-3, TIMP-1 were significantly lower in electro-acupuncture group compared with acupuncture and warm-needle acupuncture group (P < 0.05 or P < 0.01).Conclusions Acupuncture,warm-needle acupuncture and electro-acupuncture all can regulate cartilage metabolism-related factors and effectively delay cartilage degeneration,especially warm-needle acupuncture.

Abstract:Objective To investigate the therapeutical effect of Ginkgo Biloba extract (GBE) on different ages of parkinsono Biloba e (PD) mice that induced by MPTP. Methods Sixty adult mice (8 weeks) and sixty older mice (32 weeks) were randomly divided into 6 groups: negative group, model group, positive group, high, medium and low dose GBE group. PD animal model were built through injecting with MPTP and then treated with different drugs accordingly. The limb motor function of mice were tested by using pole-climbing, hanging and swimming test. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the content of malondial dehyde (MDA) in the brain tissue were measure using spectro photography. The number of positive neurons expressing tyrosine hydroxylase (TH) in substantial nigra pars compacta and dopamine transporters (DAT) in striatum were detected by using the method of immunohistochemistry. Results 1. The motor function score, the activity of GSH-Px, and the mean optical density of TH and DAT in brain tissues of model group of 8 weeks significantly decreased compared with negative group, while the content of MDA was significantly raised, showing the model of PD was successfully established. The function score, the activities of SOD and GSH-Px, the mean optical density of TH and DAT in brain tissues of the GBE treatment group significantly raised compared with the model group, while the content of MDA was significantly decreased. 2. The motor function score, the activity of GSH-Px, and the mean optical density of TH and DAT in brain tissues of model group of 32 weeks significantly decreased compared with negative group, while the content of MDA was significantly raised, showing the model of PD was successfully established. The function score, the activities of SOD and GSH-Px, the mean optical density of TH and DAT in brain tissues of the GBE treatment group significantly raised compared with the model group, while the content of MDA was significantly decreased. Conclusions The preventive medication of GBE shows a protective effect on mice of different ages with PD induced by MPTP.

Abstract:Objective To optimize the background of the congenital cataract in rats, the blood pressure, body temperature and weight of rats were measured and correlation analysis. Methods We took three kinds of rats(cataracts rat,microphthalmos cataracts rat and normal rat)as subjects, each kind with half males and females was divided into three groups by weight, each of which contains 30 rats. We used the Multi-channel physiological signal acquisition system for test. Results Blood pressure has no significant difference among the three segments of male and female rats(P > 0.05). With the increase of body weight,blood pressure and body weight formed a relevant relationship gradually.In the same male strain,there was significant difference between the standard weight(207 g) and the aging weight(494 g)(P < 0.01). In the same body weight range, the temperature difference between male and female rats was significant(P < 0.01),and the male's temperature was lower than the female. Conclusions The changes in blood pressure,body weight and temperature of cataract rats conformed to the physiological characteristics of rats,which provide a theoretical basis for the use of catatacts rats in the future

Abstract:Objective To establish a practical animal model of chronic metabolic syndrome (MS) and to investigate the correlation and pathological mechanism between hyperuricemia and glucose/lipid metabolism. Methods High fat purine diet + oteracil potassium were used to establish the MS model. Quail serum uric acid and triglyceride were tested every one/two weeks and other parameters were determined at the end of the procedure along with histopathology examination. Results Uric acid and triglyceride levels in serum were markedly enhanced by the long-term stimulation of high fat purine diet + oteracil potassium. Glucose tolerance test showed that the peak value of glucose and the area under curve in model group were higher than the others, as well as the insulin levels. Meanwhile, the modeling agents increased aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen, creatinine and induced liver, kidney and artery injuries. These injuries were partially alleviated by bezafibrate or febuxostat. Conclusions The results of the present study suggest that high fat purine diet + oteracil potassium could induce a simple, stable and durable animal model of chronic metabolic disorders.

Abstract:Objective To evaluate the efficacy and renal histopathological observation, microsphere was used as an embolic agent for unilateral renal artery. Methods Eight beagle dogs are included in this study. According to the experimental design, eight beagle dogs were randomly divided into 4 groups with each group 2 dogs. At the first, fourth, twelfth and twenty-sixth week after embolization, Each dog was divided into embolization group and normal control group by self-control. The left renal arteries were selected as the target organs for embolization with microspheres. Two dogs were chosen for the examination of angiography and DSA to observe the change of the embolized kidneys,respectively. Then the dogs were sacrificed and histological evaluation of the embolized kidneys was performed. Results Gross examination showed the size of the kidney was gradually smaller than the contralateral kidney at the fourth,twelfth and twenty-sixth week after embolization. The kidney was light colored, uneven. The surface of the kidney was gradually rough, uneven. Renal capsule was hard to separated from the kidney. Light microscopy showed that microspheres may still be seen in the small branches of arcuate artery at the fourth, twelfth and twenty-sixth week after embolization. Arterial wall hyaline, peripheral arterial fibrosis. With the time of embolism extended, A variety of degenerative changes such as atrophy, degeneration, renal infarction can be seen in the early stage; Many progressive changes such as renal tubule regeneration, granulation tissue and remodeling and other pathological changes can be seen in later stage. There is almost normal glomerulus and renal tubule structure in the embolismic kidney.Conclusions Microsphere has an specific effect of embolization on unilateral renal artery.

Abstract:Objective To study the influence of footshock (FS) -stress duration on anxiety-like behavior in the elevated plus-maze and open-field tests. Methods Rats (24) were randomly divided into 2 groups: the control group and the FS group. In the FS group, the anxiety model of rat was produced by inevitability footshock (5 minutes in total, once a day; 0.8mA, lasted for 10 seconds, 20 seconds interval) combined with separation. Anxiety-like behavior was evaluated by elevated plus maze and open-field tests after 7 and 14 days of FS-stress. Results After 7 days and 14 days of FS-stress, FS group showed significantly enhanced anxiety-like behavior and reduced locomotion indicated by total entries as compared to the control group (P < 0.01). In the open field test, the entrance times and the retention time in the central zone, the total movement distance, the numbers of rears and grooming in rats in the FS group were lower than those in the control group, which showed lower locomotion and exploratory activity in the FS groups (P < 0.01,P < 0.05).Compared to 7 days of FS-stress, both CT% (the percentage of time spent in the close arm) and CE% (the percentage of entries to the close arm) were reduced after 14 days of FS-stress showing the decreasing anxiety (P < 0.05), as well as the total movement distance and the numbers of rears and grooming (P < 0.05). The reduced locomotion and exploratory activity and self-regarding in the FS group demonstrated the enhanced depression-like behavior in rats.Conclusions Chronic FS-stress contributes to lower locomotion and exploratory activity in rats and leads to anxiety-like behavior in rats. The increased duration of chronic FS-stress may lead to the behavior change from anxiety-like behavior to depression-like behavior, and 7-days is better than 14-days in making the anxiety model in rats.

Abstract:Objective Investigating the key factors in constructing animal model of osteosarcoma. Methods 24 C3h mice (C3h/HeJ type 6, C3h/HeN type 18 at 4-8 weeks old) were randomly divided into 2 groups: A group: Subcutaneous inject 0.2 mL of 5×10⁷cells/ml mL LM8 suspension in lateral armpit before treated with depilatory creams; B group: subcutaneous inject 0.2 mL of 5×10⁷cells/mL LM8 suspension in lateral armpit before treated with shaver, and the tumor growth was recorded. Results The rate of A was 30% (4/12), and the rate of B was 100%. Conclusions Injected techniques and the way of post handle play important role in constructing osteosarcoma model.

Abstract:Diethylnitrosamine (DEN) is highly toxic to both human and animals, even the parenteral or oral administration of the smallest quantities results in severe liver damage. Due to the robustness of the induced hepatic alterations, the application of DEN in rodents has become an attractive experimental model for studies aimed at understanding the pathogenetic alterations underlying the formation of liver cancer. The liver injury induced by DEN in mice can cause liver cirrhosis and tumors, which makes it an ideal animal model to analyze the pathogenesis and process of human hepatocellular carcinoma. However, some studies have indicated that the carcinogenic effect of Diethylnitrosamine may be related to the different genetic background, gender, age and other factors. Therefore, this article reviews the relevant literatures, in order to establish a stable, high success rate, low mortality DEN model, and to provide references for different purposes.

Abstract:Embryonic Stem cell Test(EST) is an alternative testing method for testing embryotoxicity and developmental toxicity which have been validated by the European Centre for Validation of Alternative Methods(ECVAM).Under the promote of new technology, EST method has been refined by many ways including gene transfection, utilizing human-derived stem cell, increase the assay throughput, integrating test endpoints and quantifying the test indexs,which enhanced scientific relevance and testing efficiency.Within the principle of integration testing strategy(ITS),conducting pre-validation tests and standardized experiments on a large scale of chemicals, the new EST method as a individual test or a part of integrated test systerm is promising to apply on the research of lead drugs and the embryotoxicity prediction and mechanism study of chemicals.