Table of Content

28 August 2022, Volume 39 Issue 4

Previous Issue   

Analysis of Blood Routine, and Biochemical Markers, and Sex Hormones of Marmota Himalayana Artificially Feeding at Low Altitudes

NIE Jinqiu, ZHU Qingfeng, YAN Bin, ZHU Bin, ZHOU Shunchang, YANG Dongliang, WANG Yuanzhi, WANG Baoju

2022, 39(4):  1-6.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 001

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Objective The purpose of this research is to provide reference for research on health management, breeding and even industrialization and standardization of Marmota himalayana under artificial feeding conditions at low altitudes. Method In this study, we tested the blood routine markers, biochemical markers and sex hormones of Marmota himalayana under low-altitude artificial feeding conditions in Xinjiang Uygur Autonomous Region. Result There was no significant difference in blood routine indexes between male and female marmots. The hemoglobin (Hb) level of marmots aged 3 years and over was significantly higher than that of marmots under 3 years old (P<0. 05) , while the platelet ( PLT) level of marmots under 3 years of age was significantly higher than that of marmots aged 3 years and over (P<0. 05) . The platelet ( PLT) level of marmots reared for less than 1 year was significantly higher than that of marmots reared for more than 1 year ( P< 0. 001) . The serum total bilirubin ( TB) and direct bilirubin (DB) levels of female marmots were significantly higher than that of males (P<0. 05) , while the serum urea nitrogen (BUN) levels of males were significantly higher than females (P<0. 05) . The serum total protein ( TP ) , albumin ( AL ) and creatinine ( CREA) levels of marmots aged 3 years and above are significantly higher than those of marmots under 3 years old ( P < 0. 05 ) . There were no statistically significant differences in the blood biochemical test indicators between the different feeding time groups. The serum estradiol level of female marmots aged 3 years and older was significantly higher than that of females under 3 years of age (P<0. 05) , and the serum estradiol level of female marmots aged 3 years and older was significantly higher than that of males of the same age ( P<0. 001) . The serum estradiol level of female marmots during the same feeding time was higher than that of males (P<0. 05) . Conclusion Marmota himalayana that grow at high altitudes can well adapt to low-altitude artificial feeding. This study has a good reference value for the use of Marmota himalayana in low-altitude areas for scientific research.


Effect of Huaiqin Ointment on perianal edema and wound healing in mouse model with hemorrhoids via NF-kB signal pathway

ZHAO Junqing, FAN Rongrong , DOU Longtao, FANG Lei , LI Shaorong , PENG Ji , WANG Qian

2022, 39(4):  7-12.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 002

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Objective To explore the effect and potential mechanism of Huaiqin ointment on perianal edema and wound healing in mouse model with hemorrhoids. Method 30 ICR mice were used to establish the mouse model with hemorrhoids via injecting acetic acid to perianal region, then mice were divided into control group and treatment group randomly. Mice in control group were treated with low concentration potassium permanganate cleaning, and mice in treatment were treated with low concentration potassium permanganate cleaning plus applying Huaiqin ointment. After 7 days treatment, the degree of perianal edema and wound healing in perianal region was analyzed. The hs-CRP, IL-6, TNF-α, hemoglobin levels in plasma or perianal tissues were measured by ELISA or qPCR. HE staining and immunofluorescence were used to detect the inflammatory cell infiltration in perianal tissues. Western blot was used to measure the phosphorylation of IkBα and intranuclear NF-kB p65 contents in perianal tissues. Result The degree of perianal edema, wound healing, the blood hs-CRP, the cytokines ( IL-6, TNF-α) in blood or perianal tissues, hemoglobin in perianal tissues of treatment group ameliorated significantly compared to control (P<0. 05) . HE staining and immunofluorescence showed that the inflammatory cell infiltration in perianal tissues in treatment group was significantly fewer than control group ( P<0. 05) . Western blot showed that the IkBα phosphorylation and content of p65 in nuclear of treatment group was also significantly lower than control ( P < 0. 05 ) . Conclusion Huaiqin Ointment treatment attenuates inflammation response and perianal edema, promotes wound healing, of which mechanism is involved in NF-kB signal pathway.


Improvement and evaluation of a rat model of postpartum depression induced by hormone withdrawal following hormone-simulated pregnancy

WANG Xiaofei , YAN Yu , ZHAO Zhan , HE Mu , WANG Jingxuan , WU Junzhu, YANG Huaijie

2022, 39(4):  13-17.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 003

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Objective To improve the method of establishing a postpartum depression rat model after hormone withdrawal following hormone-simulated " pregnancy" and evaluate the model. Method 25 SD female rats were randomly divided into normal group ( n = 10) , experimental group ( n = 15) . Qualified animals were selected after ovariectomizing bilaterally in the experimental group, which received daily injections of the hormones estradiol 2. 5 μg and progesterone 4 mg for the first 16 days, estradiol 50 μg for the next 7 days. In the normal group, no treatment was performed, and the same amount of olive oil was injected for 23 days. In the first week after modeling, the behavioral tests were conducted through Open Field Test ( OFT) and Forced Swim Test ( FST) . Result Compared with the normal group, the experimental group exhibits significantly shortened horizontal movement distance in the OFT and increased immobility time in the FST. The difference was statistically significant (P<0. 05) . Conclusion By improving the method of establishing model, a model of postpartum depression rat after hormone withdrawal following hormone-simulated " pregnancy" was successfully established, and a reliable evaluation method was found.





Analysis on the predictive effect of excitability in early screen of guide dogs

DONG Jianyi , LI Changyi , ZHANG Min, BAI Jing , WANG Fujin, ZHOU Zijuan, WANG Jingyu

2022, 39(4):  18-22.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 004

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Objective To analysis on the predictive effect of excitability in early screen of guide dogs, four items related to excitability, PLAY, chase test, passive test and distance test, were evaluated in temperament test of the trained dogs. Method The subjects were 92 Labrador Retrievers dogs from China Guide Dog Training Centre, guide dogs( n = 59) and elimination dogs( n = 33) . By 4 excitability tests ( PLAY, Chase, Passive, Distance) and 9 behavioral variables elated items ( interest, grabbing, tug-of-war, chase average, grabbing average, moving range, moving time, posture change, exploration) were tested in Labrador Retrievers. We were trying to analyze the difference between performances of guide dogs and elimination dogs, between dogs of different sex , between guide dogs and elimination dogs of the same sex. Result Between guide dogs ( n = 59) and eliminated dogs ( n = 33) . According to the results of statistical analysis, The 4 excitability tests average score were significantly difference between guide dogs and eliminated dogs ( t = 2. 31, P < 0. 05 ) . The passive test showed significant difference between guide dogs and eliminated dogs ( t = 2. 05, P<0. 05) . Between male ( n = 54) and female ( n = 28) ,no significant difference was detected for the scores of the 4 excitability tests between male dogs and female dogs ( t = 0. 03,P>0. 05) , the passive test in male dogs was significantly higher than that in eliminated dogs( t = 2. 56, P<0. 05) . While, Between female guide dogs ( n = 24) and eliminated dogs ( n = 14) ,The distance test in female guide dogs was significantly higher than female eliminated dogs ( t = 2. 20, P<0. 05) . In the 9 behavioral variables elated items, moving range test ( t = 2. 25, P<0. 05) , exploration test( t = 2. 20, P<0. 05) in male guide dogs was significantly higher than that in female eliminated dogs. Analysis between male guide dogs ( n = 35) and eliminated dogs ( n = 19) , no significant difference was detected for the scores of the 9 behavioral variables elated items between male guide dogs and male dogs. Conclusion This study indicated that potential that the assessment of excitability is meaningful in early screen of guide dogs.

Effect of IL-6 / JAK2 / STAT3 signaling pathway on proliferation of gastric cancer

HUANG Yalin, NIU Shiwei , GONG Hongxia, SU Yun, ZHANG Han, ZENG Yuanding, LI Feifei

2022, 39(4):  23-27.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 005

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Objective To study the effect of IL-6 / JAK2 / STAT3 signaling pathway on the proliferation of gastric cancer. Method MFC gastric cancer tumor-bearing mice models were established and randomly divided into model group,AG490 blocker group, and Stattic blocker group, each with 12 mice, and another 12 healthy mice were selected as the blank group. After the intervention,the tumor tissue was taken to test the weight and calculate tumor inhibition rate;ELISA method to detect IL-6 content in mouse serum; Immunohistochemical method was used to detect IL-6,JAK2,STAT3,c-Myc and Cyclin D1 protein expression in tumor tissues;Western blot was used to detect IL-6,JAK2,p-JAK2,STAT3,p-STAT3,c-Myc and Cyclin D1 protein expression in tumor tissues;RT-qPCR method to detect IL-6,JAK2,STAT3,c-Myc and Cyclin D1 mRNA expression in tumor tissues. Result Compared with the control group,the IL-6 content in the serum of the model group was increased (P<0. 01) ;Compared with the model group, serum IL-6 content,tumor mass,the protein content of IL-6,STAT3,p-JAK2,p-STAT3,c-Myc and Cyclin D1,and the mRNA content of IL-6, JAK2, STAT3, c-Myc and Cyclin D1 decreased ( P < 0. 05, P < 0. 01 ) . Conclusion Blocking the IL-6 / JAK2 / STAT3 signaling pathway can affect the expression of downstream target proteins,thereby inhibiting the proliferation of gastric cancer. 

Establishment and Application of Fluorescent Quantitative PCR assay for Pigeon Circovirus

LI Xiaobo, WANG Ji, WANG Shasha, LI Wei, QIN Xiao, HUANG Zongwen, YUE Bingfei, WANG Shujing, FU Rui

2022, 39(4):  28-32.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 006

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Objective To establish and apply TaqMan probe fluorescent quantitative PCR method for pigeon circovirus( PiCV) . Method By comparing the sequences of various isolates of PiCV published by NCBI, selecting the conservative sequence of polymerase gene to design primers and probes, establishing a fluorescent quantitative PCR method for PiCV, and verifying the specificity and sensitivity of the method ; the plasmid standards which concentration was 3. 48×10 5 and 3. 48 ×10 4 copies/ μL, each concentration was tested in 6 parallels, the test was repeated 3 times, the coefficient of variation intra-assay and inter-assay was calculated to verify the repeatability and stability of the method ; The method was used to detect 40 samples of pigeon liver, spleen, lung, bursa of Fabricius and 36 samples of pigeon cecum collected from a breeding farm in Beijing, verifying the capability of the method in practical application. Result The established PiCV fluorescence quantitative PCR method has a linear relationship between the Ct value and the plasmid concentration in the range of 3. 48×10 7 -3. 48×10 1 copies/ μL. The standard curve Slope is - 3. 381, the R 2 value is 1, and the amplification efficiency is 97. 61%. The lowest detectable concentration of PiCV is 34. 8 copies/ μL. Compared with the conventional PCR method , the detection sensitivity is 2 orders of magnitude higher; There was no amplification curve for porcine circovirus type 2, avian adenovirus type I and III, avian leukemia virus, reticuloendotheliosis virus as templates; repeated detection of plasmid standards at 3. 48×10 5 and 3. 48×10 4 copies/ μL showed that the coefficient of variation intra-assay was less than 1. 63%, and the coefficient of variation inter-assay was less than 7. 23%. The established fluorescence quantitative PCR method was used to detect pigeon tissue samples. The positive rates of liver, spleen, lung, bursa of Fabricius and cecum were 97. 5%, 100%, 100%, 100%, and 97. 2% respectively. The chi-square test showed no significant difference ( P > 0. 05) in virus content of each tissue. Conclusion The established PiCV FQ-PCR method can effectively detect PiCV in pigeon tissues, providing technical support and basic data for the formulation of relevant microbial standards for experimental pigeons.

Effects of unsaturated fatty acids on the expression of P21, P53, P13K and PKB / Akt in liver cancer transplanted mice

ZHU Xichun, ZHANG Yongxing , YU Farong

2022, 39(4):  33-38.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 007

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Objective To establish SCID mice tumor model for screening new antitumor drugs. Method Liver cancer tissue was transplanted subcutaneously into severe combined immune deficiency ( SCID) mice. When the transplanted tumor grew to 6-7 mm, fifty SCID tumor-bearing mice were divided into unsaturated fatty acid low, medium and high dose groups [50 mg / ( kg·d) , 100 / ( kg·d) , 200 mg / ( kg·d) ] . Pentafluorouracil ( 5-FU) group [100 mg / ( kg·d) ] and model group [ distilled water 2 mL / ( kg·d) ] were administered continuously for 21 d. The transplanted tumor was removed to measure its quality and volume, and the tumor growth was compared. The protein expression levels of P21, P53, P13K and PKB / Akt in tumor tissues were determined by enzyme-linked immunosorbent assay (ELISA) . Result The weight or volume of liver cancer transplanted tumor in low, medium and high dose of unsaturated fatty acids groups and 5 -FU group was significantly lower than that in model group (P<0. 01) . The level of P21 was significantly higher compared with model group (P<0. 05, P<0. 01) ,the levels of P53, P13K and PKB / Akt were significantly lower than those in model group (P<0. 05, P<0. 01) . Conclusion Unsaturated fatty acids can inhibit the growth of liver cancer transplanted tumor, The possible mechanism is related to up-regulation of P21 level, down-regulation of P53, P13K, PKB / Akt levels, and inhibition of PI3K-Akt signal transduction pathway activity.

Curcumin reduces seizures in mouse

WU Lianlian, QIN Ying, HU Ankang

2022, 39(4):  39-43.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 008

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Objective To explore the effect and mechanism of curcumin on seizures in mice. Method 60 C57 / BL6 male mice were randomly divided into normal group, epilepsy model group and curcumin treatment group, with 20 mice in each group. Racine grade score of seizures and the latency of seizures in each group were observed and recorded. The expressions of GFAP and IBA-1 in hippocampal CA3 region were detected by immunofluorescence staining, and the expression levels of hippocampal IL-1β, NLRP3, caspase1 were detected by Western blot. Result Racine score of seizures in curcumin treatment group was significantly lower than that in epilepsy model group(P< 0. 05) , and the seizure latency was significantly prolonged ( P < 0. 05 ) . Compared with the control group, the expression of GFAP and IBA-1 in hippocampal CA3 region in the model group was significantly increased ( P < 0. 01) , glial cells were significantly activated, and the expression levels of hippocampal IL-1β, NLRP3, caspase1 were upregulated significantly ( P < 0. 01 ) . Compared with the model group, the number of GFAP and IBA-1 positive cells in hippocampal CA3 of curcumin group were decreased dramatically( P< 0. 05) , and the expression levels of hippocampal IL-1β, NLRP3, caspase1 were decreased notably ( P < 0. 05) . Conclusion Curcumin can reduce the severity of seizures, which may be through the regulation of NLRP3 / caspase1 to inhibit hippocampal inflammation. 

Study on the efficiency of tubal transplantation in mice with different genetic backgrounds

DU Miao, CHEN Fanghua, ZHAN Dawei

2022, 39(4):  44-46.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 009

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Objective Effects of different numbers of embryos transferred in mice of different genetic backgrounds on the litter rate. Method The 2-cell embryos were transplanted into pseudopregnant mice by fallopian tube transfer technique. Then, Statistical analysis was performed on the litter size of pregnant animals. Result Compared with the closed colony, the transplant efficiency of the inbred strain animals is lower, the average litter rate is 37. 5%, and the closed colony is 45%. Immunodeficient animals have higher requirements for the number of embryos to be transplanted, and mice with 11-15 cells have the highest litter rate, 63. 5%; In this study, mice with different genetic backgrounds have different mean litter rate: C57BL / 6J ( 49. 8%) > KM ( 45. 8%) > ICR (44. 1%) > NOD-SCID (35. 6%) ; > BAL B / c -nu ( 29. 3%) . In this study, it was found that mice of different genetic backgrounds required different numbers of embryos to achieve higher litter rate. ICR, C57 and NOD-SCID mice need to transfer 11-15 embryos, KM mice need to transfer 16-30 embryos; BALB / c-nu mice need to transfer ≤10 embryos. Conclusion Tubal transplantation differs in mice with different genetic backgrounds. The litter rate of mice of the same genetic background varies with the number of transferred embryos.

Establishment of a Method for Pulmonary Function Test System in Rats and Influence of Environmental Factors and Body Weight 

WANG Wendi, SUN Qi, DU Huibo, ZHAO Zhenao, ZHAO Zigang, NIU Chunyu

2022, 39(4):  47-50.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 010

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Objective To develop a stable and reliable method for pulmonary function testing in rats. Method Before the experiments, the small animal pulmonary function test system was calibrated. Lung function indices of rats were measured by the system after tracheal intubation. The influence of environmental factors and body weight on lung function indices was evaluated. Result Environmental factors, including changes of air pressure or airflow, can influence the pressure sensor and alter the waveform. The rats with heavier body weights (340-370) g showed significantly increased functional residual capacity ( FRC ) , dynamic compliance ( Cdyn ) , inspiratory resistance (RI) , forced expiratory volume in 100 ms ( FEV100) , forced vital capacity ( FVC) and mean mid expiratory flow (MMEF) when compared to the rats with lower body weights ( 280 - 300) g. However, the body weights of rats did not show significant effects on inspiratory resistance ( RI) . Conclusion An easy and reliable method was successfully established for pulmonary function testing in rats. This experiment should be performed in a stable environment, and the body weights between different groups should be consistent. 

Material and detection method of experimental rabbit thymus

FANG Hui , TIAN Yuefeng

2022, 39(4):  51-54.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 011

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Objective To observe the developmental involution process of the rabbit thymus and the precautions when harvesting it. Method Eight large ear white rabbits were obtained and randomly divided into a blank group and an immunosuppressive model group with four rabbits in each group. The immunosuppression model was prepared with cyclophosphamide in the model group, which was injected intraperitoneally at 60 mg / kg for 7 consecutive days. Twenty days after the end of modeling, thymus tissues were harvested for HE staining. Result The general morphology showed that the thymus was mostly bilobed on the left and right, with a hard texture and more tough than adipose tissue. Among them, the thymus of the blank group was mostly light pink, and the thymus of the cyclophosphamide model group was mostly white. HE result showed that the thymus was structurally intact, the corticomedullary boundary was clear, and the cortex was densely lymphocytic; in immunosuppression model group, the thymus lost its normal structure, the corticomedullary boundary was unclear, the cortex was lymphopenic, and the surrounding adipose tissue was more. Conclusion The immunosuppression model caused by cyclophosphamide would produce irreversible destruction to the morphological structure of thymus tissue in the organism.

Application of Self-made Hooks in Bilateral Ovariectomy of Rats

ZHANG Ying, SUN Xiaoyong, ZHANG Ailin, TIAN Feng

2022, 39(4):  55-58.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 012

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Objective Explore more efficient surgical approaches to bilateral ovarian excision in experimental rats. Method Ordinary ovarian extraction method is to use ophthalmic tweezers to remove the ovaries from the abdominal cavity and then ligate the uterine horns and the ovaries bilaterally, and then use ophthalmic scissors for resection, and finally inject them with penicillin intramuscularly. This method has a small amount of blood oozing, which is prone to intra-abdominal infection. In order to improve the success rate of surgery, after many explorations, this experiment made a tool dedicated to exploring and fixing the ovaries, changing bilateral ligation to unilateral ligation, penicillin intramuscular injection to intraperitoneal injection. Result Comparison experiment showed that the new method was simpler and easier to operate, it was not easy to bleed during the operation, and the success rate of the postoperative operation was higher. Conclusion The new experimental method is simpler to operate and has a higher postoperative success rate.

Progress in experimental animals with chronic circadian disturbances and its pathogenesis

LIU Weiwei, YU Yang, WEN Deliang

2022, 39(4):  59-63.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 013

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In modern lifestyles, people are affected by shifts, light pollution, lack of sleep and irregular mealtimes, causing chronic dissynchronization between internal clocks and the environment, which can lead to disrupted circadian rhythms. When circadian rhythms are disrupted or maladjusted, they can have adverse health consequences and increase the risk of diseases such as cancer, cardiovascular disease or metabolic disorders. In recent years, a wide range of animal models of circadian rhythm disorders have been established by means of core gene knockout, chemical induction, physical and dietary intervention, etc. In this paper, we summarized the commonly used animal models of circadian rhythm disorders, and briefly summarized the pathogenesis of circadian rhythm disorders.

Advances in Animal Models of MPTP Induced Parkinson’ s Disease

LYU Haiyan, HOU Guangyu, ZHU Mei

2022, 39(4):  64-68.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 014

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Parkinson’ s Disease ( PD) is a neurodegenerative disease, mainly manifested by motor symptoms such as resting tremor, muscle rigidity, bradykinesia and postural instability, accompanied by mental and cognitive disorders and autonomic dysfunction and other non-motor symptoms. The degeneration and death of dopamine (DA) neurons in the substantia nigra is the main feature of PD , and its pathological sign is the formation of Lewy bodies. People have established a variety of animal models to study different aspects of PD. Disease models can be used base on neurotoxin or gene method in non-human primates ( NHP ) , rodents, zebrafish, C. elegans, drosophila and other animals, the models prepared are to simulate the pathological changes of PD, and have their own advantages and limitations. At present, the PD model induced by 1-methyl-4-phenyl-1, 2, 3, 6- tetrahydropyridine (MPTP) is more and more widely used in medical research.

Research Progress of Single Nucleotide Polymorphismin Genetic Monitoring of Inbred Laboratory Mice 

SHANG Yanjiao, ZHOU Xiaojun, YUAN Zheng, QIU Yefeng

2022, 39(4):  69-73.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 015

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With the rapidly advancesin molecular biology technology, single nucleotide polymorphisms ( SNP ) detection technology has been constantly developed. As the third generation of genetic marker, SNP detection has become an important means of genetic monitoring in mice. SNP detection is carried out at the DNA level, thus its accuracy depends on annotation of mice genome. In this paper, the progress of genomics and SNP in genetic monitoring of inbred mice were summarized.

Research Progress of Drug-induced Zebrafish Cardiovascular Disease Models

REN Zhetan , XIE Yuyao , LIU Anqi

2022, 39(4):  74-78.  DOI: 10. 3969 / j. issn. 1006-6179. 2022. 04. 016

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This article describes the method for various drug-induced heart failure models in zebrafish. Zebrafish have been shown to mimic human cardiac disease due to their similarity to mammalian hearts, ease of genetic manipulation, and rapid and acclimatized development. The article describes the advantages and disadvantages of zebrafish as a humanized animal model in heart failure disease models, discusses the use of different drugs to build zebrafish models to reveal the specific mechanism of heart failure and cardiac remodeling, and concludes that zebrafish fish model will contribute to the in-depth study of the pathophysiological mechanism of human cardiovascular disease, and provide new prevention and treatment ideas for heart failure.