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    28 June 2025, Volume 42 Issue 3
    Comparison of SNP Molecular Marker and Biochemical Marker in Genetic Monitoring of BALB / c Mice
    ZHAO Lan, LIU Wei, ZHOU Jiaqi, WEI Jie, WANG Hong, ZHANG Xinyan, LI Huan, FU Rui, YUE Bingfei
    2025, 42(3):  1-8.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 001
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    The similarities and differences between the current national standard biochemical marker method and the Sanger sequencing detection mouse SNP molecular marker method in the evaluation of genetic quality of BALB / c mice were compared and analyzed, so as to provide a reference for the genetic quality control of inbred mice. Methods The genetic quality of the same batch of BALB /
    C mice was tested using 14 conventional biochemical marker loci and 34 common genetic analysis loci of SNP.The result evaluation, variation detection, chromosome coverage, animal welfare, operation  convenience and method repeatability were compared between the two methods. Results Six BALB / cmice with No. 2, 5 and 6 variants were detected by both methods, but only homozygous variants at 4 loci were detected by biochemical marker, while homozygous variants at 9 loci and heterozygous variants at 3 loci were detected by SNP molecular marker. Further comparison showed that SNP marker method was superior to biochemical marker method in chromosome coverage, animal welfare, operation convenience and method repeatability. Conclusion SNP marker method has more advantages in genetic quality control evaluation of inbred mice than the current national standard biochemical marker detection
    method, which can be used as a standardized genetic quality control technology for inbred experimental animals.
    Validation of Inactivation of Minute Virus of Mice and the Evaluation of the Capability Verification Results of Nucleic Acid Detection
    WANG Ji, WANG Shasha, QIN Xiao, LI Wei, WANG Shujing, LI Xiaobo, WANG Hong, FU Rui
    2025, 42(3):  9-17.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 002
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    To determine the effective temperature and time for inactivation of minute virus of mice (MVM) to ensure the biosafety of the laboratory’ s samples. Through the implementation of MVM nucleic acid detection capability verification plan, we can understand the inspection capability of relevant testing institutions of laboratory animals in China, so as to improve the quality detection level and
    detection capability of laboratory animals in our country. Methods MVM with known virus titer was inactivated by water bath heating at different temperatures and different times. Some of the inactivated viruses were used for virus titer detection and some were used for nucleic acid detection by fluorescent PCR. The fecal suspension of MVM nucleic acid negative mice was taken as negative samples. The  inactivated virus was added to MVM nucleic acid-negative mouse fecal suspension to prepare positive samples. After passing the uniformity and stability test, the samples will be randomly numbered and distributed to the participating units. NIFDC will summarize and analyze the result submitted by the participating units. Results When the inactivation temperature reaches 100 ℃ and the inactivation time does not exceed one hour, the virus can be effectively inactivated. Meanwhile,the variation coefficient of nucleic acid test result in batches is less than 5%,and the difference is not significant. The satisfaction rate of the participants was 96. 4% ( 27 / 28) . Conclusion
    When the inactivation temperature reaches 100 ℃ and the inactivation time does not exceed one hour, the MVM nucleic acid sequence remains intact, and the nucleic acid detection result do not affect the experiment. There were 96. 4% of the participating laboratories had the capability of MVM nucleic acid detection, and 3. 6% needed to be improved. There are still a small number of laboratories that need to be improved in terms of operational norms and the writing of original records.
    Establishment and Preliminary Application of a TaqMan Real-time Quantitative PCR Assay for Detecting Canine Pathogenic Escherichia coli
    WU Jingqi , LIN Wei , YANG Jiamei , GUO Yingcheng , LI Liheng , ZHAO Lili
    2025, 42(3):  18-22.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 003
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    To overcome the limitations of traditional detection method for Canine pathogenic Escherichia coli ( CPEC) , a rapid, sensitive, and specific TaqMan probe-based real-time quantitative PCR ( qPCR) method was established for the accurate diagnosis of CPEC. Methods
    A qPCR detection method targeting the conserved virulence gene eaeA of CPEC was developed. Standard curves were established using serial dilutions to determine the linear range and the limit of detection ( LOD ) .Specificity and repeatability were evaluated. Forty-five clinical samples were used to compare and validate the qPCR method with conventional PCR. Results The established qPCR method exhibited a good linear relationship in the standard curve (R 2 = 0. 99) , with the equation Y = -3. 406x+40. 14. The limit of detection was 10 copies/ μL. No cross-reactivity was observed with other common pathogens.Repeatability experiments showed that both intra-assay and inter-assay coefficients of variation were below 3%. In the detection of clinical samples, the qPCR method identified 15 positive cases, and the consistency between the two methods reached 93. 3%. Conclusion This study successfully established a highly sensitive and specific TaqMan probe-based qPCR detection method, which can provide effective technical support for the early diagnosis and epidemiological monitoring of canine CPEC infections.

    Establishment and Application of a Warning Detection Method for Mouse Hepatitis Virus Barrier Facilities Based on RAA-CRISPR / Cas13a Technology
    CAI Heqing , ZHANG Xuliang , CHEN Xiaojuan , WANG Lie , DAI Fangwei , LI Wei
    2025, 42(3):  23-29.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 004
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    To establish a rapid MHV monitoring method based on environmental media for early pathogen warning in laboratory animal barrier environments. Methods Conserved sequences of MHV were analyzed to design specific RAA primers and crRNA compatible with the LwCas13a reaction system. After optimization, a RAA-CRISPR / Cas13a detection system for MHV was developed, with validation of its specificity and sensitivity, followed by application in animal environmental dust sample detection. Results A rapid MHV detection method based on Recombinase aided amplification ( RAA) combined with CRISPR / Cas13a technology was established for dust samples from laboratory animal habitats. Specificity tests demonstrated no cross-reactivity with common murine viruses including Sendai  virus, Reovirus type 3, Pneumonia virus of mice, Encephalomyelitis virus, and Murine norovirus. The method showed high specificity for MHV detection with a superior sensitivity of 10 copies/ μL detection limit. Clinical validation using PCR as reference involved parallel testing of 160 random dust samples, revealing complete consistency between both methods and identifying 26 antigen-positive samples.Conclusion The RAA-CRISPR / Cas13a detection system based on environmental dust provides efficient and sensitive technical support for pathogen surveillance in laboratory animal facilities.

    Establishment and Application of a Duplex RT-qPCR Detection Method for Group A Rotavirus and Seneca Virus
    LIU Xing, LUO Tingyu, ZHANG Yu, LI Kaili, YIN Bozhao, LI Changwen, XIA Changyou, GAO Caixia
    2025, 42(3):  30-37.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 005
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    To establish a dual RT-qPCR detection method for porcine rotavirus A ( PoRVA) and seneca virus A ( SVA) and conduct preliminary applications. Methods By comparing the sequences of PoRVA and SVA strains, conserved regions of the PoRVA NSP3 gene and SVA VP1 gene were selected to design primers and probes, respectively, and a dual RT-qPCR detection method for PoRVA and SVA was established. Subsequently, the specificity and sensitivity of the method was confirmed.Recombinant plasmid standards with concentrations ranging from 10 6 to 10 3 copies/ μL were serially diluted 10 - fold. Each concentration was tested in triplicate and the tests were repeated three times.Intra- and inter-assay coefficients of variation ( CV) were calculated to evaluate the repeatability and
    stability of the method. The established dual RT-qPCR method was compared with the detection method recommended by national standard “ GB / T 34756 - 2017” and industry standard “ SN / T 5486 - 2022” .Additionally, 15 pig fecal samples and 35 pig anal swab samples were tested to evaluate the practical applicability of the method. Results The established dual RT-qPCR method showed a linear relationship between Ct values and plasmid concentrations in the range of 109-101copies/ μL. The standard curve for PoRVA had a slope of -3. 085, an R 2 value of 0. 993, and an amplification efficiency of 110. 956%. The standard curve for SVA had a slope of -3. 085, an R 2 value of 0. 993, and an amplification efficiency of 110. 935%. The minimum detectable concentrations for both PoRVA and SVA were 101 copies/ μL. No cross-reactivity was observed with other seven common swine pathogens. Repeated testing of plasmid standards at concentrations of 106 to 103 copies/ μL showed intra-assay CVs of less than 0. 15% and inter assay CVs of less than 1. 50%. The established dual RT-qPCR method and the national standard and industry standard method were used to detect the positive rate of 50 samples. The result showed that the positive rate of PoRVA and SVA were 100% (50 / 50) and 66% (33 / 50) for the dual RT-qPCR method, respectively. The positive rate of PoRVA and SVA were 100% (50 / 50) and 24% ( 12 / 50) for “GB / T 34756- 2017” and “ SN / T 5486 - 2022” method, respectively. The positive concordance rate reached 100%. Conclusion The dual RT-qPCR detection method established in this study can effectively detect PoRVA and SVA, providing technical support for disease detection and routine monitoring in swine populations.
    Effects of Mannitol on S100β Protein and TLR4 / NF-κB Pathway in Rat Brain after Cardiopulmonary Resuscitation
    FENG Kai , CHENG Aibin , MEN Xiuli , WANG Jianjun , BU Xuan , LI Shuang , BAI Jing
    2025, 42(3):  38-45.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 006
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    Objective To investigate the effects of different usage frequencies and total doses of mannitol on cerebral arterial blood flow, serum S100β protein concentration, and the protein and mRNA expression of the TLR4 and NF-κB in brain tissue after cardiopulmonary resuscitation ( CPR) in rats,and to explore the role of mannitol in brain injury after CPR. Methods A total of 120 male Wistar rats were used to establish a cardiac arrest and CPR model, and were randomly divided into 3 groups ( n = 40, sham operation group, model group, and mannitol intervention group) . The model group and mannitol intervention group underwent the same model. After successful resuscitation, the mannitol intervention group received 20% mannitol at different time points.The sham operation group only underwent intubation through the trachea and catheter placement via femoral artery and vein puncture. Laser Doppler flowmeter was used to monitor the changes in cerebral arterial blood flow in the rats at 6, 12,18, and 24 h after surgery. Tissue specimens were collected from the rats at 6, 12, 18, and 24 h after surgery for detection of serum S100β protein concentration using ELISA method. H&E staining was used to observe the pathological changes in rat brain tissue, and Western blot was used to detect the protein
    expression levels of TLR4 and NF-κB. Real-time PCR was used to detect the mRNA expression levels of TLR4 and NF-κB. Results
    The cerebral arterial blood flow in the model group was significantly lower than that in the sham operation group after the restoration of spontaneous circulation, and the difference was statistically significant ( P < 0. 05) . In the mannitol intervention group, the cerebral arterial blood flow in rats increased after mannitol administration 5 times during the 6, 12, and 18 h post-surgery periods, and decreased after 7 times of mannitol administration ( 24 h) , with no significant difference compared to the model group.Swelling of hippocampal neurons, increased nuclear staining and shrinking, increased glial cell proliferation, and disordered arrangement of neurons were observed in the model group. In the mannitol intervention group, the arrangement of neurons in the cerebral cortex and
    hippocampal tissue was more orderly than that in the model group at 12 h post-surgery, with reduced cell necrosis. A large number of deep-stained and shriveled neuronal nuclei were observed at 24 h post surgery. A significant increase in the S100β protein content in rat serum was observed in both the model group and the mannitol intervention group at 12 h post-surgery, compared to the sham operation group.The S100β protein content in the model group significantly increased at 6, 12, 18, and 24 h post-surgery (P<0. 05) , while in the mannitol intervention group, the S100β protein content significantly decreased at 6, 12, and 18 h post-surgery (P<0. 05) compared to the model group, with no significant difference at 24 h post-surgery. Compared to the sham operation group, the protein and mRNA expression levels of TLR4 and NF-κB in hippocampal tissue of rats in the model group significantly increased at 6, 12, 18,
    and 24 h post-surgery (P<0. 05) . However, in the mannitol intervention group, the protein and mRNA expression levels of TLR4 and NF-κB in the hippocampal tissue significantly decreased at 6, 12, and 18 h post-surgery (P<0. 05) , and the protein and mRNA expression levels of TLR4 in the hippocampal tissue were significantly decreased at 24 h post-surgery ( P< 0. 05) . Conclusion Moderate amounts of mannitol can improve cerebral arterial blood flow, reduce serum S100β protein, and decrease the protein and mRNA expression levels of TLR4 and NF-κB in brain tissue after CPR, thereby protecting brain tissue and alleviating secondary brain injury. Excessive mannitol may cause cerebral edema, worsen intracranial hypertension, and lead to more severe secondary brain injury.
    Effect of Mononoside on Oxidative Stress in Rats with Heart Failure after Myocardial Infarction
    FEI Yihuan , LIU Tingting , XU Zhidong, SUN Fangling , TIAN Xin , ZHENG Wenrong, ZHU Zixin , WANG Yufeng , WANG Wen
    2025, 42(3):  46-53.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 007
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    Objective To investigate the effect of morroniside on oxidative stress in heart failure rats induced by ligation of the left anterior descending coronary artery. Methods The rat model of heart failure was established by ligating the left anterior descending coronary artery and inducing myocardial infarction. The rats that survived the procedure were then divided into six groups: a model group, a low dose group of morroniside (60 mg / kg) , a medium-dose group of morroniside (120 mg / kg) , a high-dose group of morroniside (240 mg / kg) , captopril group (100 mg / kg) and the sham operation group was only threaded without ligation. Following a modelling period of 3 hours, the drugs were administered by gavage according to the corresponding drugs and dosage, once a day for a period of eight weeks. Hematoxylin eosin (HE) staining was used to detect the histopathological changes of the rat heart. The levels of SOD and MDA in the plasma of rats in each group were detected by WST-1 method and TBA method,
    respectively. Finally, Western blot analysis was used to detect the expression of SIRT1 and SIRT3 proteins in rat myocardium. Results
    Following a period of eight weeks, the result of HE staining demonstrated that, in comparison with the sham operation group, the myocardial cells in the model group exhibited a state of disarray, accompanied by the presence of broken and necrotic muscle fibers, as well as inflammatory cell infiltration. In comparison with the model group, the myocardial structure of rats in the morroniside group and captopril group was evidently enhanced and the infiltration of inflammatory cells was mitigated. The result of the SOD detection showed that the plasma SOD concentration in the model group was significantly lower than that in the sham operation group (P<0. 001) . In contrast, the morroniside group exhibited a significant increase in SOD concentration (P<0. 05,P<0. 01) , as did the Captopril group increased significantly ( P < 0. 01) . The result of the MDA test demonstrated that the concentration of MDA in the model group was significantly higher than that in the sham operation group (P< 0. 01) . In contrast, the morroniside group exhibited a significant decrease in MDA concentration (P<0. 05,P<0. 01) , and that in the Captopril group demonstrated a comparable decrease ( P<0. 01) .The result of the Western blot analysis demonstrated that the expression levels of SIRT1 and SIRT3 proteins in the model group were significantly lower than those in the sham operation group ( P<0. 05) . Compared with the model group, the expression level of morroniside in the High-dose group was significantly increased (P<0. 05) , and there was an increasing trend in the captopril group, though this was not statistically significant. Conclusion Morroniside may improve the oxidative stress injury in rats with heart failure subsequent to myocardial infarction by increasing the concentration of SOD in plasma, decreasing the concentration of MDA and Up-regulating the expression levels of SIRT1 and SIRT3 proteins, thereby reducing myocardial injury.
    Performance Avaluation of Self-developed Biosafety Individually Ventilated Cages
    WANG Xiwei , YU Chengwen, CHEN Hongyan , LIU Huairan , JIN Zhimin , WANG Wei , XIA Changyou
    2025, 42(3):  54-58.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 008
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    ObjectiveTo explore the detection methods for various indicators of biosafety independent ventilation cages ( IVCs) and evaluate the performance indicators of self-developed biosafety mouse IVCs.Methods According to the detection methods of RB / T199 - 2015 and DB23 / T 2057. 1 - 2017,various performance indicators of a certain imported biosafety type IVC ( IVC1) and a self-developed biosafety
    type IVC ( IVC2) were tested, and their safety was determined. Results All indicators of the prototype IVC and control IVC meet the requirements of national standards, the detection results of airflow velocity,air cleanliness, falling bacteria, noise, illumination,temperature and relative humidity are relatively consistent between the two IVCs.However, compared with IVC1, IVC2 had certain differences in
    pressure difference, ventilation frequency, and airtightness indicators. Conclusion The detection method has excellent operability and can be used for on-site inspection of biosafety mouse IVC. The self-developed biosafety mouse IVC has excellent performance, safety and reliability, and can be used for thebreeding of highly pathogenic animals in high-level biosafety laboratories.
    Protective Effect of Dicoumarol on Pathological Cardiac Hypertrophy Induced by Stress Overload
    ZHOU Siyi , LI Wei , LIU Jiayi , ZHANG Xiaojing , QU Weiyi , ZHANG Peng
    2025, 42(3):  59-65.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 009
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    Objective This study aimed to investigate the protective effects of dicoumarol on pathological cardiac hypertrophy induced by stress overload, thus providing scientific evidence for its potential therapeutic role. Methods We established pathological cardiac hypertrophy models in vitro using primary cardiomyocytes and in vivo using mice, administering dicoumarol and evaluating its protective effects against cardiac hypertrophy. Isoproterenol stimulation was employed to induce cardiomyocyte hypertrophy in the primary cell culture. The cells were categorized into control and treatment groups. Immunofluorescence staining was conducted after 24 hours of stimulation to evaluate cellular area. In the mouse model, pathological cardiac hypertrophy was induced via aortic constriction. Mice were divided into solvent control and treatment groups, with each group comprising 9 mice. The treatment group received dicoumarol at a dosage of 10 mg / kg every day. Four weeks post-surgery, cardiac structure and function were evaluated using echocardiography, while the degree of cardiac hypertrophy and fibrosis was  assessed via histopathological staining. Results In the in vitro model, dicoumarol significantly attenuated isoproterenol-induced cardiomyocyte hypertrophy.In the mouse model of myocardial hypertrophy, dicoumarol administration markedly ameliorated pressure overload-induced cardiac dysfunction, myocardial hypertrophy, and fibrosis compared to the solvent control group. Conclusion Dicoumarol exhibits protective effects against pressure overload-induced pathological myocardial hypertrophy and fibrosis, thereby improving cardiac function.
    Investigation on the Patterns of Estrous Cycles and Dynamics of Sex Hormone Levels in Apodemus peninsulae
    WANG Zi’ an , LUO Sanpin , JIN Weihan , ZHANG Yiyun , ZHOU Xingxuan , ZHOU Jiazheng , ZHOU Zijuan , DONG Jianyi , CHEN Jun , WANG Liang , WANG Jingyu
    2025, 42(3):  66-73.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 010
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    Objective This experiment aims to explore the estrous cycle of Apodemus peninsulae and the changes in sex hormone levels at different stages of the estrous cycle. Methods Select female Apodemus peninsulae during the breeding period, and use a combination of external genital observation and vaginal smear method to determine the estrous cycle of Apodemus peninsulae. Extract serum from different stages of the estrus cycle of Apodemus peninsulae and the concentrations of estrogen, progesterone, and luteinizing hormone were measured using enzyme-linked immunosorbent assay ( ELISA) . At the same  time, hematoxylin and eosin (HE) staining was used to observe the morphological changes of the uterus and ovaries of Apodemus peninsulae at different stages of the estrous cycle. Finally, the uterine and ovarian tissues of Apodemus peninsulae were extracted, and the differential expression of sex hormone related receptor genes at different stages of the estrous cycle was analyzed using qPCR technology to explore their regulatory role during the estrous cycle. Results The estrous cycle of female Apodemus peninsulae lasts 4-7 days, with an average estrous period of 0. 97 days and an average interestrous period of 2. 41 days. Estrogen and luteinizing hormone levels were significantly higher during estrus than in the post-estrus and interestrous phases ( P < 0. 001 ) , while progesterone levels were significantly higher during interestrous phase compared to pre-estrus and estrus ( P < 0. 001) . The significant morphological and structural differences in the uterus and ovaries during the estrous cycle reflect the critical regulatory role of cyclic fluctuations in sex hormone levels on the morphological and structural changes in the reproductive organs of Apodemus peninsulae. Conclusion Estrous Cycle of Apodemus peninsulae was clearly divided into four distinct stages. The result revealed consistent morphological, hormonal, and structural changes across different phases.
    Expression of Cre Recombinase in Intestinal Epithelial-specific Cre Transgenic Mice
    TANG Yuling , LI Site , LI Zixuan , QIAN Hong’ an , WANG Jin , WU Wenqing , QIU Yefeng
    2025, 42(3):  74-79.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 011
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    Objective To investigate the expression of Cre recombinase in a kind of new intestinal epithelial-specific Cre transgenic mouse. Methods
    The expression of Cre recombinase in Vil1-Cre mouse could be investigated by detecting the expression of the reporter gene YFP, of which the Rosa26-YFP fluorescence reporter gene expression could be induced by Cre recombinase. Results In adult, Vil1-Cre mouse expressed Cre recombinase in the whole intestinal epithelium, including duodenum, jejunum,ileum, and colon. Meanwhile, a few scattered cells expressing Cre recombinase were observed in other tissues such as the kidney, pancreas, testis and ovary. During embryonic development, Cre recombinase is already expressed in the midgut at embryonic day 10. 5 ( E10. 5 ) , just one day later than the expression of endogenous villin protein at E9. 5. Besides, there is also a certain expression of Cre recombinase in the septum transversum, genital ridge and mesonephric tubule at E11. 5 and E12. 5.Conclusion The Vil1-Cre mouse showed a perfect expression pattern of Cre recombinase throughout the intestinal epithelium with high specificity and activity, which can be used to construct an intestinal epithelial-specific gene modified mouse model for related study.
    Study on Mouse Model of Liver Yin Deficiency Syndrome in Alcoholic Liver Injury
    FENG Guangxu, JIA Lan, LIU Jiaqi, HU Jinxi, FENG Yingtong, WANG Jingxia
    2025, 42(3):  80-87.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 012
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    Objective Based on the mouse model of chronic alcoholic liver injury induced by Lieber DeCarli liquid alcohol feed, intragastric administration of thyroxine to mice to make a model of liver yin deficiency syndrome of alcoholic liver injury. The model was explored from two aspects: thyroxine concentration and experimental cycle. Methods ICR mice were randomly divided into blank group,PTH 0. 16 g / kg +15 d group, PTH 0. 32 g / kg +15 d group, PTH 0. 64 g / kg +15 d group, PTH 0. 16 g /kg +30 d group, PTH 0. 32 g / kg +30 d group, and PTH 0. 64 g / kg +30 d group, with 10 mice in each group. The mouse model of liver yin deficiency syndrome of alcoholic liver injury was prepared by gavage with Lieber-DeCarli alcohol liquid feed and different concentrations of thyroid hormone in different cycles. After 15 or 30 days, the state of mice in each group was observed, body weight, body temperature, heart rate were measured, serum transaminase activity, cyclic nuclear adenosine, liver lipid, antioxidant enzyme activity and cytokine levels in liver tissue were measured, and the liver was  taken for HE pathological examination. Results Compared with the blank group, the mice in PTH 0. 32 g / kg +30 d group had significantly reduced body weight, increased body temperature, accelerated heart rate, dull hair, fatigue, irritability, dry stool, yellow urine and other symptoms(P<0. 05) . The levels of serum ALT, AST, cAMP and MDA、TC、TG、IL-6 in liver tissue increased significantly ( P< 0. 01, P< 0. 05) . The levels of cGMP in serum and SOD、IL-10 in liver tissue decreased significantly ( P<0. 01,P< 0. 05 ) . Lipid deposition and hepatocyte necrosis occurred in the liver. Conclusion Successfully established a mouse model of alcoholic liver injury with liver yin deficiency syndrome.
    Long Term Animal Experimental Study on a New-style Mitral AnnuloplastyRing Made of a hyperelastic Nitinol
    JIA Liujun , JIANG Xin , CHEN Zhuo , YANG Liu , DUAN Xuejing , HE Ting , ZHANG Qi , WANG Xuemin , FAN Junying , ZHANG Xue , YUE Guangxin
    2025, 42(3):  88-93.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 013
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    Objective We aim to evaluate the effectiveness and safety of a " new-style " cut-out mitral annuloplasty ring made of a hyperelastic nitinol through long-term in vivo animal experiments. Methods These " new-style" annuloplasty rings and Edward annuloplasty ring as control were implanted into healthy Small Tailed Han sheep, the cardiac function, the mitral valve and the heart morphology, the integrity, the local inflammation, the endothelialization and the thrombus of these " rings" , the hematolysis, the infection and the liver and kidney function were detected at post implantation day 90 and 180 through  echocardiography, pathology, complete blood count and blood chemistry. Results Echocardiography showed that there were similar effects these two-type rings on the cardiac structure and function, and the mitral valve shape. Autopsy, revealed that all rings tightly coupled with the mitral ring of receptors without breaking, cracking, or deformation, the circumferential fiber around the rings were thin and full, but without thrombus.Histopathology found revealed that both types of formed rings had complete fibrous capsules, with only mild local inflammatory reactions around them.Scanning electron microscope discovered endothelialization was complete. The blood tests showed that there were no significant changes in the blood routine and liver and kidney functions of all individuals compared to before the operation.Conclusion This" new-style " mitral annuloplasty ring made of a hyperelastic nitinol continued functioning post implanting in Small Tailed Han sheep, it is in no way inferior to Edward annuloplasty ring.
    Improved Method for Intratracheal Lipopolysaccharide Administration to Establish a Mouse Model of Acute Respiratory Distress Syndrome
    YANG Luyu, CAO Zhimin , WANG Jiali , HAO Dongxia , ZHOU Jie , YE Huan
    2025, 42(3):  94-101.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 014
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    Objective To improve the success rate of ARDS disease model, we explored the optimal condition for constructing the model through endotracheal instillation of LPS, eventually providing a technical support for the basic research of ARDS. Methods A total of 116 C57 / BL6N mice were included in this research.We explored the optimal administration conditions by comparing the drug distribution and actual modeling effects of different body positions ( slant position, supine position) and LPS volumes ( 30 μL / 20 g, 60 μL / 20 g, 90 μL / 20 g ) . Further, different anesthesia methods were compared according to the physiopathological status of mice. The above optimized conditions were validated by ARDS modeling. Results Compared to the supine position, LPS instilling to mice in a slant position can distribute more evenly across lung lobes. A dosage of 60 μL LPS / 20 g body weight can meet the modeling requirement, such as thickening of alveolar walls and increasing of neutrophils according to  H&E staining. Compared to the control group, significant differences were observed in injury score,vascular permeability and systemic inflammation(P<0. 001) . Last but not least, intraperitoneal injection of anesthesia exacerbates inflammation in mice and affects blood oxygen saturation ( P < 0. 01 ) .Conclusion C57 / BL6N male mice (6-8 weeks old, 20 g body weight) were used for establishing an ARDS disease model. Isoflurane inhalation anesthesia was selected before surgery, which was performed in a slant position. The optimal volume of LPS was 60 μL / 20 g ( body weight) for inducing ARDS.
    Tribromoethanol by intraperitoneal injection was not recommended for anesthesia, due to side effects such as extra inflammatory burden and reduction of blood oxygen saturation.
    Investigation of the Anesthetic Effects of the Brimonidine-Chlorpromazine  Combined Anesthetic in Rabbits
    LI Xin, YU Xiaomeng, CHEN Bin, WANG Xiaolin, WANG Xiaohui
    2025, 42(3):  102-108.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 015
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    Objective To develop a compound anesthetic by proportionally combining brimonidine and chlorpromazine, evaluate its anesthetic effects on rabbits via intravenous injection, and conduct a comprehensive assessment of the compound anesthetic. Methods A dose-response screening experiment was conducted to determine the optimal formulation of the brimonidine-chlorpromazine ( BL) compound anesthetic.
    After administering 1 mL / kg intravenously, the depth and duration of anesthesia were observed in three dose groups of rabbits, and the optimal dose of BL was selected for further experiments. The BL regimen was compared with the dexmedetomidine-chlorpromazine (DL) regimen. Hemodynamic parameters, including blood pressure, heart rate, and body temperature, were monitored during anesthesia. The analgesic effects of both regimens were evaluated based on the animals’ responses and heart rate changes before and after pain stimulation. Additionally, the effect of brimonidine on increasing the pain threshold was assessed using a laser stimulation test. Results
    In the dose-response screening,both the high-dose group (0. 6 mg / mL brimonidine and 0. 5 mg / mL chlorpromazine) and the mediumdose group ( 0. 4 mg / mL brimonidine and 0. 5 mg / mL chlorpromazine ) exhibited longer anesthesia maintenance times
    compared to the low-dose group ( 0. 2 mg / mL brimonidine and 0. 5 mg / mL chlorpromazine) , with no significant difference between the high-dose and medium-dose groups. The medium dose was chosen as the optimal dose for BL intravenous anesthesia. The anesthesia induction time for the BL group was ( 2. 4 ± 1. 5) min, and the anesthesia maintenance time was ( 33. 6 ± 5. 8 ) min.During anesthesia, heart rate decreased, while blood pressure remained relatively stable, and body temperature showed a slight decrease at 75 and 90 min, all of which were superior to the corresponding indicators in the DL group. The BL regimen effectively inhibited mild to moderate pain within 20 min of administration. The average percentage increase in pain threshold for the high-dose, medium-dose, low
    dose, and control groups of brimonidine was ( 81. 43 ± 15. 52 ) %, ( 47. 81 ± 11. 04 ) %, ( 32. 99 ±6. 14) %, and ( 5. 24 ± 6. 80 ) %, respectively. Conclusion The depth and duration of anesthesia provided by the BL regimen are comparable to those of the DL regimen. However, the BL regimen has a lesser impact on cardiovascular dynamics and body temperature and can more effectively suppress mild to moderate pain. Intravenous brimonidine increases the pain threshold in a dose-dependent manner. The brimonidine-chlorpromazine
    compound anesthetic demonstrates favorable sedative and analgesic properties, making it suitable for surgeries involving mild to moderate pain stimulation.
    High Quality Development of Laboratory Animals Based on Enhancing Drug Testing Capabilities
    HE Zhengming , CHEN Zhenwen , SUN Yansong , CHEN Hongyan , DONG Qinghua , GONG Wei , LYU Longbao , LIANG Chunnan
    2025, 42(3):  109-113.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 016
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    Based on a comparative analysis of the quality requirements for experimental animals in the Chinese Pharmacopoeia (2020 edition, three parts) and national standards, this paper elaborates on the differences and limitations between the two.With the aim of improving drug testing capabilities, it explores the problems and solutions in the diversity of laboratory animals genetic resources, the application of gene modified animals, as well as animal experimental ethics and biosafety. The purpose is to provide reference for promoting the development of drug testing technology, and for ensuring drug quality and ensuring medication safety for the people.
    Research Progress on the Construction and Evaluation of Mouse Models of Pelvic Inflammatory Disease
    CHEN Yuhang , YU Ting , HUANG Ying
    2025, 42(3):  114-120.  DOI: 10. 3969 / j. issn. 1006-6179. 2025. 03. 017
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    Selecting ideal animal models of pelvic inflammatory diseases is an important basis for the study of pelvic inflammatory diseases, and scientific and systematic model evaluation is the key to further study. In this paper, the pathogenesis, modeling principle, modeling method, modeling evaluation and TCM syndrome evaluation of PID rat model were comprehensively expounded and analyzed, so as to
    provide reference for the related research of pelvic inflammatory diseases.