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Effects of a Chinese traditional medicine, Huoxue Xiaoyi Fang, on embryo development after in vitro fertilization in mouse models of endometriosis

Yangyang CUI ; Weiwei SUN ; Ruihua ZHAO ; Tiansi WU

Chinese Journal of Comparative Medicine.2015;(8):31-36,43. doi:10/3969.j.issn.1671.7856.2015.008.007

Objectives To evaluate the effect of a Chinese traditional medicine , Huoxue Xiaoyi Fang , on the embryo development after in vitro fertilization ( IVF ) in mouse models of endometriosis .Methods To observe the quantity of oocytes including the average number of superovulation , fertility rate, 2-cell and 4-cell embryo cleavage rates at different stages in vitro fertilization, and morulation rate among the traditional Chinese medicine group (TCM group), endometriosis model group ( EM model group ) and sham-operated group .To explore the influence of endometriosis and the effect of Huoxue Xiaoyi Fang on the quantity of oocytes and embryonic development after IVF in the mouse models of endometriosis.Results All the average number of superovulation , fertility rate, 2-cell and 4-cell embryo cleavage rates in the three groups showed significant differences (P<0.05), and the average number of superovulation was in an order of sham operated group >TCM group>model group .There were no significant differences in the morulation rates in the three groups (P>0.05).There were significant differences among the IVF rates of the three groups (P<0.05), and to be in an order of sham operated group >TCM group>model group .There were significant differences in the final morulation rate among the three groups (P<0.05).Conclusions Endometriosis may influence the quality of embryonic development of EM model mice, including reduced average number of superovulation , fertility rate, and embryo cleavage rate .The Chinese traditional medicine Huoxue Xiaoyi Fang can improve the the quantity of embryonic development after IVF .

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Alzheimer’ s disease-like lesions may occur in and accompany the high-sugar high-fat diet-induced rat models of metabolic syndrome

Li NIU ; Dewu HAN ; Ruiling XU ; Xin ZHOU ; Chongxiao QU

Chinese Journal of Comparative Medicine.2015;(8):27-30,43. doi:10.3969/j.issn.1671.7856.2015.008.006

Objectives To explore the relativity between Alzheimer ’ s disease ( AD)-like lesions and metabolic syndrome models induced by high-sugar high-fat diet in rats.Methods Forty-eight Sprague Dawley rats were randomly divided into 2 groups.The control group (fed with normal diet, 12 rats) and high sugar and high fat group (fed with high-sucrose and high-fat diet, 12 rats) continuously for 12 months.At the end of 6, 9 and 12 months of the experiment , we observed the animal body weight and visceral fat weight .The blood lipid levels , blood glucose and MS-related biochemical parameters were determined . The brain tissues were examined by histopathology . The characteristic AD molecules hippocampus Aβand Tau were detected using ELISA and Western blotting to confirm the presence of AD lesions in the brain.Results Compared with the normal control group , the body weight and visceral fat weight of the rats in the high-sugar high-fat groups were significantly increased; the levels of TG , FPG, LDL, HOMA-IR and hippocampus Aβ,phosphorylated Tau (p-Tau) were higher, but the level of HDL was decreased (P<0.05 for all).The histopathological examination revealed inflammatory cell infiltration in the brain tissues .Conclusions Characteristic AD-like lesions may occur and accompany the rat models of metabolic syndrome , induced by high-sugar high-fat diet, and provide a new idea for the construction of Alzheimer ’ s disease animal models .

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Research progress of the relationship between autophagy andβ-amyloid peptide in Alzheimer’ s diseese

He JIN ; Rong WANG

Chinese Journal of Comparative Medicine.2015;(8):68-71,75. doi:10.3969/j.issn.1671.7856.2015.008.014

Autophagy is an important pathway in the body to remove damaged cell organelles and dysfunctional proteins to maintain protein equilibrium and intracellular homeostasis by the lysosomal system .As a degenerative disease of central nervous system , the main feature of Alzheimer ’ s disease is abnormal protein aggregation .However , the role of autophagy in the pathogenesis of Alzheimer ’ s disease is not fully clear yet .The aim of this paper is to take β-amyloid peptide as a starting point to explore the relationship between autophagy and β-amyloid peptide in the pathogenesis of Alzheimer’s disease.

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Comparison of different special staining techniques of chondrocytes and their application values

Fei YU ; Hui ZENG ; Hongyan YU ; Ming LEI ; Hao YUAN ; Deming XIAO

Chinese Journal of Comparative Medicine.2015;(8):58-61,75. doi:10.3969/j.issn.1671.7856.2015.008.012

Objective To compare the advantages and values of several special staining methods of chondrocytes . Methods Twelve 7-day old healthy C57BL/6J mice were killed to obtain the cartilage tissue of the knee joint in order to isolate the chondrycytes .Type II collagen was used to assess the chondrocytes .Then the chondrocyte climbing slices were prepared.The materials were fixed, and HE staining, Safranin O-fast green staining, SA-β-gal staining and immunohistochemical staining of Type II collagen were performed and compared .Results HE staining showed clear morphology of the chondrocytes .The cell nuclei were stained blue and the cytoplasm was pink .Safranin O-fast green staining showed that the nuclei were pink and the cytoplasm green .SA-β-gal staining showed that the aging cells were green while the young cells were colorless .Immunohistochemical staining of type II collagen showed the distribution of type II collagen and they were stained brown while the cell nuclei were blue .Conclusions HE staining and safranin O-fast green staining can provide more information than the other staining techniques .SA-β-gal staining is useful in the analysis of aging chondrocytes .Immunohistochemical of type II collagen can be used to study type II collagen .

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Investigation of pathogen infections in Mongolian gerbils raised in a conventional facility

wei Fang DAI ; tao Jiang DU ; sang Sha ZHOU ; Yu LV ; Fei GUI ; ming Xiao SONG ; feng Xiao CHU

Chinese Journal of Comparative Medicine.2017;27(11):15-18,31. doi:10.3969.j.issn.1671-7856.2017.11.004

Objective To investigate the pathogen infections of Mongolian gerbils raised in a conventional facility,and to provide a basis for the establishment of local standards for pathogen detection in Mongolian gerbils. Methods A total of 16 species of bacteria,11 species of viruses and 8 species of parasites were detected in 30 gerbils raised in a conventional facility, according to the national standards of microorganism and parasite detection in mice and rats. Results Gerbils raised in this conventional facility were infected with lymphocytic choriomeningitis virus(a positive rate of 6. 7%), sendai virus(3. 3%), pneumonia virus of mice(100. 0%), reovirus type III(6. 7%), mouse encephalomyelitis virus(10. 0%), mycoplasma spp.(6. 7%), Tyzzer's organism(6. 7%)and Helicobacter spp. (56.7%),according to our antibody detection results. Meanwhile,the detected positive rate of Pasteurella pneumotropica was 3.3%,Staphylococcus aureus 10.0%,Escherichia coli O115 a,C,K(B)6.7%,Tritrichomonas muris 100.0% and flagellates 100.0%. Conclusions The results of our study provide a reference for the establishment of classification standards for gerbils according to their pathogen and parasite infections.

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Correlation between cognitive development and levels of dopamine and 3,4-dihydroxyphenylacetic acid in the hippocampus in 80-day-old neonatal rats born of fear-impaired pregnant rats

min Xin LI ; ping Li YANG ; jiao Hai WAN ; hui Jiang ZHU ; lin Jun HOU ; hong Xiang ZHAN

Chinese Journal of Comparative Medicine.2017;27(11):10-14. doi:10.3969.j.issn.1671-7856.2017.11.003

Objective To investigate the correlation between cognitive development and levels of dopamine(DA) and 3,4-dihydroxyphenylacetic acid(DOPAC)in the hippocampus of 80-day-old neonatal rats born of fear-impaired pregnant rats. Methods The pregnant rat model of fear-impaired kidney was established by watching other rats shocked with electricity. The cognitive development of the neonatal rats was assessed by Morris water maze test at 80 days after birth. The brain microdialysis samples of the right hippocampus were collected using a stereotaxic instrument, and the levels of DA and DOPAC were determined by HPLC-ECD. Results Compared with the control group, the mean escape latency of the 80-day-old neonatal rats in the model group was increased. Their swimming speed was slower. The 20% time spent in the peripheral zone became longer, and the frequency of platform crossing was decreased, showing significant differences(P< 0.05). The levels of DA and DOPAC in the extracellular fluid of the hippocampus of the 80-day-old neonatal rats in the model group were decreased at each time points of perfusion, showing a significant difference(P <0.05). The level of DA, a kind of monoamine neurotransmitter in the hippocampus, was positively correlated with the mean escape latency,the frequency of passing the platform of the 80-day-old rats,while was negatively correlated with the 20% time spent in the peripheral zone, with a significant correlation(P < 0.05). In addition, there was a positive correlation between the DOPAC level and the mean escape latency, the frequency of passing the platform(P< 0.05). Conclusions Fear-impaired pregnant rats can affect the spatial learning and memory ability of their 80-day-old neonatal rats,with reduction in the levels of DA and DOPAC in the hippocampus, which is closely correlated with the cognitive development of the neonatal rats.

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Effect of Osteoking on blood physiological parameters and organ indexes in female tree shrews with osteoporotic fracture

Chao WU ; Wei TANG ; bin Hong ZHAO ; tao Jin LI ; lin Jian JIAO ; Xin YUAN ; Hong ZHENG

Chinese Journal of Comparative Medicine.2017;27(11):1-5. doi:10.3969.j.issn.1671-7856.2017.11.001

Objective To evaluate the effect of Osteoking on blood physiological parameters and organ indexes in the ovariectomized tree shrew model of osteoporotic fracture(OPF). Methods A total of 30 4-month-old female tree shrews(Tupaia belangeri)were randomly divided into control group, model group and treatment group, with 10 in each group. Bilateral ovaries of the tree shrews in the treatment group and model group were removed and osteoporosis was confirmed 6 months later. Then tibial fractures were generated. From the next day of fracture,tree shrews in the treatment group were gavaged with Osteoking once every two days,and tree shrews in the model group and control group were gavaged with equal volume of sterile saline. After gavaging for 3 months,16 blood physiological parameters and 6 organ indexes of the tree shrews in each group were measured. Results Among all of the 16 blood physiological parameters,the number of white blood cells,the absolute value and percentage of lymphocytes(P < 0.01)and monocytes(P < 0.05)in the model group were significantly lower than those in the control group after 3 months. Compared with the model group, the absolute value and percentage of monocytes(P < 0.01)and lymphocytes,the number of white blood cells(P < 0.05) and the spleen coefficient were significantly increased(P < 0.05)in the treatment group. Conclusions Osteoking can improve the immunity of tree shrews with osteoporotic fracture.

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Progress in research on the susceptibility of tuberculosis

meng Meng SUN ; Jun TANG ; Chuan QIN ; jun Ling ZHAN

Chinese Journal of Comparative Medicine.2017;27(11):106-112. doi:10.3969.j.issn.1671-7856.2017.11.022

Tuberculosis(TB),a chronic infectious disease caused by Mycobacterium tuberculosis(MTB), is the outcome of the interaction between polygenes and environmental factors,which has been shown in lots of genetic studies. In recent years,the research of TB susceptibility genes and their single nucleotide polymorphism(SNP)loci has become a hot topic,but genes and SNPs exactly associated with TB susceptibility are rarely reported. What's more,some susceptibility genes are different among races, populations and geographical distributions, resulting in inconsistency, or even contrary conclusions drawn from local studies of different populations or areas. In order to obtain susceptibility genes that are widely applicable and exactly associated with TB,it is necessary to screen the genes preliminarily in the laboratory animals with gene diversity, and thereafter, verify those genes in animals or humans. The Collabrative Cross(CC)mice have the advantages of diverse and clear genetic background,which can provide a new tool for the research of TB susceptibility. This review summarizes the advances in the studies of TB susceptibility genes and their SNPs, and looking forward to the application of CC mice in these studies and its significance.

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Recent progress in research of heterogeneous nuclear ribonucleoprotein K related with tumor pathogenesis and progression

Hao HUANG ; jiu Xing YANG ; yuan Meng LI ; min Rui ZHU ; Ran GAO

Chinese Journal of Comparative Medicine.2017;27(11):100-105. doi:10.3969.j.issn.1671-7856.2017.11.021

Over the past few decades, the classification of oncogenes or tumor suppressor genes has been an important topic in cancer biology. However,it is difficult to classify some genes. Heterogeneous nuclear ribonucleoprotein K(HNRNPK)is a nucleic acid-binding protein,which is involved in the regulation of gene expression,signal transduction and many other cellular processes. In recent years, it has been found that HNRNPK is overexpressed in many types of tumors,and its overexpression is negatively correlated with the prognosis of patients,suggesting that HNRNPK may play a role as an oncogene in tumorigenesis. In contrast,however,HNRNPK has also been considered as a tumor suppressor gene in acute myeloid leukemia(AML). Therefore, in this article we summarize and discuss the recent progress in the molecular functions and regulatory mechanisms of HNRNPK in tumorigenesis and progression.

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A simplified method for isolation and culture of primary mouse hepatocytes

jun Shi FAN ; Xin LIU ; Dong YANG ; feng Yuan ZHU ; ling Yong LU ; Jiang ZHENG

Chinese Journal of Comparative Medicine.2017;27(11):75-79. doi:10.3969.j.issn.1671-7856.2017.11.015

Objective To simplify and optimize the method for isolation and culture of primary mouse hepatocytes on the basis of conventional extraction method,and to provide a reference for related research. Methods Mouse liver was reversely perfused with the isolation solution(i.e.,through the vena cava inferior in,and portal vein out),cut into small pieces and digested with enzymes. Then the hepatocytes were isolated by density gradient centrifugation and transferred into culture medium. The cell viability was detected by trypan blue staining. The purity of the hepatocytes was analyzed by flow cytometry and the cell morphology was observed with an inverted microscope. Results The hepatocytes obtained by this improved method showed high viability and purity,with typical characteristics of cell morphology. Conclusions The liver perfusion is facilitated by reversed perfusion, and the isolated hepatocytes are with high viability and purity confirmed by many times of experiments. This optimized procedure is an easy and efficient method for isolation of primary mouse hepatocytes.

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