Table of Content

28 October 2018, Volume 35 Issue 05

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H istological Observation of Digestive and Neurvous System s of Gobiocypris rarus

2018, 35(05):  1. 

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Ob jective The histological description of digestive and neurvous systems was given by systematically observation of adult Gobiocypris rarus，which provides a more complete background of differentmammal studies on the evolution of these systems， with the detailed pictures.Method Healty adult Gobiocypris rarus were fixed，decalcified，dehydrated and vertrified in order.The specimen in solid paraffin was cut into serial slices with thickness of 3μm.The slices were dyed by Hematoxylin and Eosin（H&E）staining.The section pictures of the organs in the fish were described when observation and collection through a optical microscope were finished.Result Pharyngeal teeth，as a marker divides the pharynx conncected cavity and intestine of Gobiocypris rarus.The anterior intestine is enlaged to a“Z”，while the posterior seems to be straight.The liver， in which there are distinct central veins but typical lobule，is covering the intestine.Gall bladder， next to the heart， is singly located in the front of the thoracic cavity.Pancreas or livers with exocrine pancrestic tissue has not been observed in the fish.The brain， the most important part composed the CNS of Gobiocypris rarus， whose substances of telencephhalon， diencephalon， mesencephalon， metencephalon and myelencephaloncan have been fully developed and easily observed.Conclusion There is no stomach in Gobiocypris rarus which is accorded with the characteristic of Cyprinidae fish.Distinguished differences have been found between the intestines from anterior and posterior segments.In a degree，the nervous system of the species is fully developed.

Effects of G inkgolide B on Energy M etabolism in Rats w ith M yocardial Ischem ia/Reper fusion In jury

2018, 35(05):  7. 

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Ob jective To investigate the effects of ginkgolide B on energy metabolism in rats with myocardial ischemia/reperfusion injury.Method SD ratswere divided randomly into sham group，model group，ginkgolide B low dose group （2 mg/kg）， ginkgolide Bmedium dose group （4 mg/kg） and ginkgolide B high dose group （8 mg/kg） （n=10）.Myocardial ischem ia/reperfusion injury model was established by coronary artery ligation （30 m in）and subsequent reperfusion （2 hours） in rats.Ginkgolide B was injected via tail vein twice at1 h pre-operation and the start of the reperfusion， respectively.After the end of reperfusion， the CK and LDH activities in sera were examined by chromatometry，Na+/K+-ATPase and Ca2+/Mg2+-ATPase activities in myocardial tissue were detected by phosphorus method and the myocardial infarction size（MIS） was determ ined by staining method.Result High dose of ginkgolide B significantly reduced MIS to 16.20%，comparing to 20.12%ofmodel group（P＜0.05）.The CK activities in ginkgolide B groups were 0.69，0.64 and 0.63 U/m L，respectively，which were significantly lower than 0.77 U/mL ofmodel group （P＜0.05 or P＜0.01）.Ginkgolide B（8 mg/kg） significantly reduced LDH activities to 2.20 U/m L， comparing to 2.99 U/m L of model group （P＜0.01）.Ginkgolide B（8 and 4 mg/kg） evidently increased Na+/K+-ATPase activity to 3.69 and 3.78 mmol Pi/g， respectively， comparing to 3.09 mmol Pi/g ofmodel group （P＜0.05）.Ginkgolide B significantly increased Ca2+/Mg2+-ATPase activity to 4.50，4.79 and 4.81 mmol Pi/g，respectively，comparing to 3.97 mmol Pi/g ofmodel group （P＜0.05 or P＜0.01）.Conclusion The protective effects of ginkgolide B pretreatment in myocardial ischem ia/reperfusion injury appears to be associated with the improvement of energy metabolism.

Experim ental Study of Com bined Hyperlipidem ia Rat M odel Induced by Different K inds of Diets

2018, 35(05):  11. 

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Ob jective To observe the influence of combined hyperlipidem ia ratmodel induced by different kinds of experimental diets.Method One hundred male SD rats were randomly divided into ten groups： basic diet group，and from one to nine experimental diet group， ten rats in each group， respectively.The rats of basic diet group were treated with basic diet，while the others were treated with corresponding experimental diet for 4 weeks.Then the levels of TC，TG，HDLC，LDLC，GLU were examined after treated for 2 and 4 weeks，and the ratio of liver weight and body weight， the ratio of fat weight and body weight， and histopathology changes of the liver were exam ined after treated for 4 weeks.Result Compared to the basic diet group，the experimental diet groups could decrease the average daily food intake， increase the level of TC， LDLC， the ratio of liver weight and body weight，and the degree of hepatic steatosis（P＜0.05）， but did not increase the level of GLU.Comparison among the experimental diet groups， the first experimental diet group could increase the level of TG（P＜0.05）， the fourth experimental diet group could increase the level of TC， LDLC， the ratio of liver weight and body weight（P＜0.05）， the seventh experimental diet group could Less affect the average daily food intake（P＜0.05）.Conclusion The nine kinds of experimental diets could induce combined hyperlipidemiaratmodel，the first experimental diet group could more advantageous to increase the level of TG，the fourth experimental diet group could more advantageous to increase the level of TC， LDLC and the hepatic lesions， the seventh experimental diet group were fit for rat，and less influence on weight.

Construction of HE4（WFDC2） Knockout M ouse Using TALEN Method

2018, 35(05):  17. 

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Ob jective To construct HE4 （WFDC2） knockoutmice using microinjection of transcription activatorlike effector nuclease（TALEN） mRNAs.Method TALENs-mediated precise knockout site specifically targeting mouse WFDC2 gene was designed，and the correspondent vectorswere constructed.TALENs cDNA was transcribed in vitro initially，then the F0-generation m ice were obtained from the microinjection of TALENs mRNA into the C57BL/6 Jmouse zygotes.F0 mice were characterized at genomic DNA level by using genotyping，and WFDC2 knockoutmice were confirmed.Result We designed and constructed TALENs which recognized and cut the first exon of Wfdc2， and got 24 F0 m ice， among which one had frameshift mutation， demonstrating that the WFDC2-knockoutm ice were generated by TALEN technique，and we also find that the homozygous null m ice died after birth.Conclusion WFDC2 gene knockout m ice model were successfully developed by using the combination of m icroinjection and TALENs-mediated deletion of mouse WFDC2 gene.This will be a useful model to study the WFDC2's physiological function in mouse，and could also provide the insight on the involvement of WFDC2 in certain diseases such as ovarian and endometrial cancer progression

The Effect of an Improved Sheep Vertebral Bone Defect Model for Bioactive Vertebral Augmentation Materials

2018, 35(05):  24. 

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Objective To improve a sheep vertebral bone defect model for bioactive vertebral augmentation substitutes study. Method Eighteen goats with the mass is (21. 56 ±2. 56 ) kg were selected. L1-L6 were provided by retroperitoneal approach under intravenous anesthesia. A bone void with a size of 6 mm × 10 mm was created in each of the vertebrae by adrill, and was filled by calcium phosphate cement ( CPC ) , CPC combined with autologous platelet rich plasma ( PRP) , or nothing in each 2 vertebrae randomly. Six goats were randomly executed 1 , 3 , 6 months after surgery, then the imaging and histological observation were performed. Result The operation time was 64 - 92 (75 ± 6.95 ) min, and the blood loss was 50 -110 (81.94 ± 15. 64 ) mL. 108 vertebral body specimens were harvested .There was nostatistically significant diference in volume ratio between the three groups (P >0.05 ) . Both two bone cement groups began to have different levels of bone cement absorption and bone growth from 3 months after surgery, but the intervention group was more obvious than control group. Conclusion It is simple and easy to establish the lumbar vertebrae bone defect by retroperitoneal approach, with few operative complications. Besides, long-term survival of animals and observation of filling materials can be achieved. In this study, the bone formation effect CPC is fine, and can be expanded when combined with PRP.

Effect of Gender on G lucocorticoid-induced Osteoporosis Rat M odel

2018, 35(05):  30. 

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Ob jective Study the effect of gender on glucocorticoid-induced osteoporosis（GIOP） rat model.Method The female and male rats of GIOP group were both injected dexamethasone 2.5 mg/kg intramuscularly from d1 to d21 and 1 mg/kg from d22 to d63 once every two days，while control group was injected equivalent normal saline intramuscularly.One h after the last injection，animals were sacrificed.The BMD of femur and L4 were measured by dual energy X-ray.ALP， Ca， P and Vitam in D3 of serum were determ ined.Tibia was collected for pathological examination.Result Compared with control group，the femur and L4 BMD ofmale rats in GIOP group were decreased， serum ALP increased while Ca， P and calcitonin decreased， and the score of tibia pathological examination increased.On the female rats of GIOP group， there're only femur BMD decreasing， ALP increasing and tibia pathological score increasing.Conclusion Compared with female rat，male rat is more suitable to be used to built glucocorticoid-induced osteoporosismodel.

Prevalence and Rapid Identification of Toxoplasma gondii In fection am ong Laboratory Anim als in China

2018, 35(05):  35. 

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Ob jective In order to establish a rapid identification method for Toxoplasma gondii（T.gondii）， and acquire the prevalence data on T.gondii infestation in laboratory animals in China，and provide reference for the revision of national standard， a survey was conducted.Method A total of 12 394 laboratory animals（including 290 monkeys， 425 mini-pigs， 579 dogs， 1 234 rabbits， 372 hamsters， 1 363 guinea pigs， 987 rats， 7 144 mice）came from 60 different manufactures in China between December 2012 and December 2016 were chosen as respondents.Using enzyme-linked immunosorbent assay（ELISA）， sera were tested for IgG antibody and CAg antigen of T.gondii.Polymerase chain reaction（PCR） was used to detect p30 gene of T.gondii.Direct microscopy real-time dynamic video recording technique was performed to identify the toxoplasma pathogen.Result Of 12 394 exam ined animals 4 were positive for T.gondii IgG antibody.Molecular characterization by PCR amplification of the p30 gene demonstrated the presence of T.gondii in these animals.Tachyzoites of T.gondii were detected in these animals by using directm icroscopy real-time dynamic video recording technique.Among the total 12 394 laboratory animals investigated from December 2012 to December 2016，the prevalence of T.gondii infection was 0.03% （4/12 394）.Conclusion ELISA and PCR combined with direct microscopy real-time dynamic video recording techniques can be used for the detection of T.gondii.T.gondii infection was present among laboratory animals in China.Thus，it is suggested to design reasonable screening programs for preventing animal-transmitted toxoplasmosis.New diagnostic tools， especially rapid detection technologies， which is necessary to be developed in the future studies on this important zoonotic disease.

Effects Induced by Different Cond itions on the Differentiation of Tree Shrew Bone M arrow M esenchym al Stem Cells into Osteob lasts

2018, 35(05):  46. 

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Ob jective To investigate the effects of different mediums on the differentiation of tree shrew bone marrow mesenchymal stem cells（BM-MSCs） into osteoblasts in vitro.Method Tree shrew bone marrow mesenchymal stem cells（MSCs） were isolated and cultured in vitro by density gradient centrifugation and adherent culture.The tree shrew bonemarrow mesenchymal stem cells of P3 generations were divided into 7 groups.Group A：high glucose DMEM+1 μmol/L dexamethasone+100 μmol/L vitam in C+10 mmol/Lβ-glycerophosphate；Group B：high glucose DMEM+50 ng/m L BMP-2；Group C：high glucose DMEM+80 ng/m L BMP-2；Group D：high glucose DMEM+50μmol/L Vitam in C+10 mmol/Lβ -glycerophosphate+20 ng/m L BMP-2；Group E： high glucose DMEM+1 μmol/L dexamethasone+100 μmol/L Vitamin C+20 mmol/L βphosphoglyceride； Group F： high glucose DMEM+100 μmol/L dexamethasone+100 μmol/L Vitam in C+10 mmol/Lβ-phosphoglyceride；Group G：DMEM/F12+0.1μmol/L dexamethasone+50μmol/L Vitamin C+10 mmol/Lβ-phosphoglyceride.After 18 d，stain the cells with alkaline phosphatase and alizarin red.Result The result of each group of alkaline phosphatase staining were showed positive in different degree；the result of alizarin red staining in group A，B，C，D and group E were observed significantm ineral nodules.Conclusion Group A，group B，group C，group D and group E induced differentiation mediums could induced tree shrew BM-MSCs to differentiate into osteoblasts，among which group A and group B were the most effective.

The Effect of Sub Packaged Freezing-thaw ing on Serum Biochem ical Test in SD Rats at-20℃for Cryop reservation

2018, 35(05):  52. 

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Ob jective To investigate the effect of sub packaged freezing and thawing on serum biochemical test in SD rats at-20℃for cryopreservation.Method A fter the instrument was calibrated and quality control was passed，the blood of 40 blank female SD rats was collected，stored for 30 min at 2～8℃，then centrifuged，the serum was dispensed and sealed with a sealing film，and each animalwas dispensed in 4 portions.Serum detection analysis was done under room temperature for 4 h and the resultwas the reference level.The remaining 3 sera were stored at-20 ℃ for cry preserved and were re-thawed and detected after 2 d，7 d，14 d respectively.The effects of biochemical test result at room temperature for 4 h were compared with those at-20℃for cryopreservation for different time.Result There were statistical differences of the concentration of TP，ALB，GLU and CREA in the serum biochem ical test result between the condition of freezing-thawing at-20℃for 14 d and room temperature for 4 h （P＜0.05）.There were statistical differences of the concentration of ALT， TP， ALB， GLU， UREA，CREA and TC in the serum biochemical test result between the condition of freezing-thawing at-20℃for 7 d and room temperature for 4 h （P＜0.05）.There were statistical differences of the concentration of ALT， TP， ALB，GLU and CREA in the serum biochem ical test result between the condition of freezing-thawing at-20℃for 2 d and room temperature for 4 h（P＜0.05）.There was no statistical difference of the concentration of AST and TG in the serum biochemical test result between the condition of freezing-thawing at-20℃ for 2 d，7 d，14 d and room temperature for 4 h （P＞0.05）.Conclusion The serum biochemical test result of AST and TG were stable and consistent between the condition of freezing and thawing at-20℃ for 2 d，7 d，14 d and room temperature for 4 h while the other seven result have changed.This experiment shows that m inim ize the number of serum freezethawing as few as possible if there is no special circumstance，and suggests that it is best to detect in that day otherwise itwill affect the accuracy of the test result.

The Study of Exp ression of CaM in M ice Liver after Welfare Im paired by Restrain Stress

2018, 35(05):  56. 

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Abstract：Ob jective To explore the effect of restraint stress on the expression of calmodulin in mice liver.Method Twenty three-week-old ICR rats were divided into two groups：control and rats under restraint stress.Individual weight gain and feed consumption were recorded during the experiment.After one week of continuous restraint treatment， blood samp ling were taken to measure the neuroendocrine， immunology indexes and liver function indexes，the expression of CaM and CaMKⅡ in liver were evaluated by Western Bolt.Result Compared with the control group， the individual weight gain and FER of restraint group were decreased （P＜0.01）， the organ index of spleen and thymus were significantly decreased（P＜0.01，P＜0.05）， CORT， ACTH and liver function indexes AST， ALT， ALP were significantly increased（P＜0.01，P＜0.05）.The protein expression of CaM and CaMKⅡ were much higher than those in the control group（P＜0.05）.Conclusion After welfare damaged by restraint stress， CaM and CaMKⅡ were involved in the process of welfare being impaired.

Analysis of Random Testing Results of Laboratory Anim als in Beijing Area from 2014 to 2016

2018, 35(05):  60. 

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Ob jective To provide reference to production， management and detection of laboratory animal by analyzing of the recently 3 years'test result（2014—2016） on m icrobes and genetics of laboratory animals in Beijing.Method According to national and local standard and relevant law and regulation，Beijing Adm inistration Office of Laboratory Animal organized quality monitoring test for qualified laboratory animal manufacturers.According to the 3-year data from test reports，we can analyze the quality of different kinds of laboratory animal（including mice， rats， guineas， shrews，rabbits， dogs， monkeys and minipigs） in Beijing area.Result From 2014 to 2016， there were 120，133 and 130 batches of random testing annually， while 16，17，23 unqualified batches respectively.The main reason for disqualification is the immunity level dissatisfaction（49 batches in rabbits， dogs and minipigs）.There were also 3 batches genetic variation and 1 batch Pasteurella Pneumotropica positive in mice.1 batch Salmonella positive and 1 batch reovirusⅢ antibody positive were found in guineas，and 1 batch Salmonella positive was found in dogs.Conclusion Quality monitoring of laboratory animal is the important means for the quality management，and it can provide scientific data for the production and management of laboratory animals.

Effect of Raspberry Extract on Anxiety of Nicotine With d rawal in Rats

2018, 35(05):  67. 

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Ob jective In the present study， we investigated the effect of raspberry on anxiety-like behavior and the levels of norepinephrine in hippocampus during nicotine withdrawal in rats.Method Sprague-Dawley rats（220—250 g）were treated with 0.4 mg/kg of nicotine or saline by daily subcutaneous injection for 7 days.24 hours after the last injection，rats in the contour group were given diazepam，the experimental group were given raspberry（100 mg· kg-1·d-1、200 mg·kg-1·d-1）， for five days.One hour after the final dose of raspberry， the anxiogenic response was evaluated using an elevated plus maze，the concentrations of norepinephrine in hippocampus were measured by high performance liquid chromatography.Result Rats undergoing nicotine withdrawal exhibited substantial anxiety-like behavior，which was characterized by the decrease in time spent in the open arms of the elevated plus maze，which were greatly attenuated by doses of raspberry.The high performance liquid chromatography analysis revealed that nicotine withdrawal significantly increased norepinephrine levels in the hippocampus.Similar to the result from the elevated plus maze test，the raspberry significantly inhibited the elevation of norepinephrine.Conclusion These result suggest that raspberry attenuates anxiety-like behavior induced by nicotine withdrawal through modulation of the hypothalamic norepinephrine system in the brain.

Discussion on the Welfare Ethics for Rats in the Use of M olar Rods

2018, 35(05):  71. 

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Ob jective The teeth of rats grow rapid ly.In order to improve the animalwelfare in rats， to promote the normal growth and development of teeth，to ensure the correctness of the tests and the result，and to determine the feasibility of the using of the Lesser Khingan Range white birch molars，to provide theoretical basis for future research on animal toys.Method We use the molar rod thatmade up with the Betula papyrifera in the Lesser Khingan Range， M illettia Laurentii， im itation wood ivory and stainless steel water mouth to explore the body weights， food consumption， viscera indexes and serum biochemical indexes in the SD rats feeding process.Result Showed that the Betula papyrifera molar rod have no significant effects on the growth and development of rats.The Betula papyrifera molar rod in the Lesser Khingan Range have practicality and feasibility，can helps rats grow and develop，improve animal welfare in rats.

Pathological Observation of Canine Ovarian Sex-Cord Strom al Tum ors

2018, 35(05):  75. 

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Objective To three kinds of canine ovarian sex-cord stromal tumors pathology diagnosis and analysis，canine ovarian sex-cord stromal tumors pathological characteristics，provide data for the diagnosis of canine ovarian sex-cord stromal tumors.Method China Agricultural University National Animal TSE Lab collected from China Agricultural University animal hospital clinical cases of ovary neoplasm， and made into slices， H.E.staining.Result Compare histopathological features of dog granulosa cell tumor， thecoma and luteoma， summarize the clinical features of ovarian sex-cord stromal tumors，for the diagnosis and differential diagnosis of this tumor in the future to provide effective information.Conclusion Granulosa cell tumor， thecoma and luteoma， for common ovarian sex-cord stromal tumors， common in older dogs.Due to tumor cell development degree is different， presents different histopathological features.

The In fluence of the Comm unal Nest on Social Recognition of C57BL/6J M ice

2018, 35(05):  79. 

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To explore the effect of early environmental enrichment on social recognition，60 adolescent C57BL/6J m ice collected from the standard nest and communal nest was tested on the basis of the habituation-dishabituation model and analyzed，compared.Both the female and male C57BL/6 Jm ice showed the ability of social recognition in the standard nest and the communal nest.And compared with in the standard nest，the social recognition of C57BL/6 Jm ice did not significantly been enhanced in the communal nest.We conclude that social recognition of the animals that did not significantly been enhanced may be because social recognition is themost basic ability，and the environment has little effect on it.In addition，the research on the communal nest would be help ful to study other higher social behavior.

Research Progress of Circu lar RNAs and Nerve System Disease

2018, 35(05):  83. 

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Circular RNA（circRNA） is one kind of closed circular RNA molecule that produced by backsplice，they are stable， conserved， diverse and exhibit tissue or developmental stage specific expression.The functions of circRNA include roles asm icroRNA（m iRNA） sponges， regulating the expression of its parent gene by cis or trans，and participating in protein translation.In addition，circRNA in the physiological process of neural development and related diseases， which is expected to become biomarker.In this review， we focus on the characteristics of circRNA molecular， formation mechanism， functions， databases in common use and its relationship with diseases of the nervous system.

The Research Progress of Anim al M odel of Leukem ia

2018, 35(05):  88. 

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Mice is an idealmodel animal for leukemia research， which provides important reference function to the leukemogenesis， the development law， the optimization of treatment and the judging prognosis.And it can also make human avoid the huge risk of human trials，being one of the effective tools in basic research.After researchers'ongoing efforts and constant innovation and optimization of establishment method，the biological properties ofm ice model has been closer to clinic and reproduced the relevant performance of clinical leukem ia，which has laid a foundation for leukemia research.This paper mainly summarizes the establishment method，research progress and identification method of leukem ia m ice model in recent years.