Table of Content

25 December 2018, Volume 38 Issue 6

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Progress and Perspective for Xenotransplantation

ZHANG Qi, WANG Jian-fei

2018, 38(6):  407-411.  DOI: 10.3969/j.issn.1674-5817.2018.06.001

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Worldwide, the number of patients needing organ transplants for end-stage organ failure is growing, howeverŁ¬ the number of organs available from donation after cardiac death/donation after brain death (DCD/DBD) or living donors is very limited. Therefore, the use of pig organs/tissues for xenotransplantation is one of the final solutions for the shortage of organs. Although tremendous progress has been made in xenotransplantation research, there are still some huge hurdlers, especially immune rejection, in the clinical application of xenotransplantation. With the continuous progress of the field, especially the development of various gene editing technologies, knocking out specific porcine genes which cause hyperacute rejection and humanizing multiple genes in pigs become possible, the survival of transplanted organs in the models of non-human primates has been significantly improved. Moreover, the application of CRISPR/Cas9 technology to remove porcine endogenous retroviruses (PERV) from the pig genome eliminates the potential risk of endogenous pathogens infecting the host after xenotransplantation, which is of landmark significance. This work reviews the barriers, progress, safety, oversight and current status of xenotransplantation.

Study on Pathological Changes and Related Mechanism Induced by Brain Injection of Serum in Rats

ZHAO Zhi-jing, DIWU Fei-hu, DENG Yi-heng, YANG Li-sun

2018, 38(6):  412-416.  DOI: 10.3969/j.issn.1674-5817.2018.06.002

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Objective To investigate the pathological changes and related mechanism induced by brain injection of serum in rats. Methods Injection of fetal bovine serum (FBS) into rat brain. The control group injected the same volume normal saline, after 48h contrast two groups survival rate, behavioral change, the measurement of brain tissue water content changes, RT-PCR detection of aquaporin-4(AQP-4) mRNA expression ; The expression of AQP-4 protein was detected by Western blot. The expression of AQP-4 mRNA was detected in serum/serum-free culture of rat astrocytes, RT-PCR, and the expression of AQP-4 protein was detected by Western blot. Results The survival rate and behavioral score of the experimental group were significantly improved compared with the control group. The edema of brain tissue around the injection site of the experimental group was shown under electron microscope, the expression of AQP-4 protein and mRNA in the experimental group was significantly higher than that in the control group (P<0.01), the expression of AQP-4 protein and mRNA in the injection site of the experimental group were not significantly different from that of the control group (P<0.05); Compared with serum-free culture group, the expression of AQP-4 protein in astrocytes of rat with serum and mRNA were significantly decreased (P<0.05). Conclusion In vitro and in vivo, it is shown that the decrease of AQP-4 expression in brain tissue, especially astrocytes may be an important molecular mechanism of secondary cerebral edema after intracerebral hemorrhage (ICH), and provide a new therapeutic target for the treatment of cerebral edema after ICH in the future.



Observation on Correlation between Insulin Resistance Index and Blood Lipid in Obese Rats

SHENG Yi-xuan, ZENG Guo-wei, YAO Liang-liang, LI Bing-tao, JIANG Li, ZHANG Qi-yun, XU Guo-liang

2018, 38(6):  417-421.  DOI: 10.3969/j.issn.1674-5817.2018.06.003

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Objective To obesrve the correlation between blood lipids and insulin resistance in obese rats induced by high-fat diet. Methods Male Sprague-Dawley rats were continuously fed with high fat diet supplemented with 36% fat energy for 12 weeks. The food intake, body weight were detected, Lee's index was calculated, serum which was collected by multiple time periods and the degrees of serum insulin, fasting blood glucose, total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol(HDL-C) and low density lipoprotein cholesterol (LDL-C) were measured, and insulin resistance index was calculated. Cluster analysis was used to screen successful insulin resistance rats and their differences in body weight, Lee's index, biochemical indicators compared to normal groups were analysed. Result Rats induced by high-fat diet developed obesity after 4 weeks with higher levels of body weight and Lee's index (P<0.01, P<0.05). Sixteen rats producing insulin resistance were selected by cluster analysis based on the 12-week insulin resistance index of obese rats. After statistical analysis, 16 rats developed insulin resistance after 8 weeks, with lower levels of TC and HDL-C as compared with the normal group (P<0.05). But there was no significant change in FBG, TG and LDL-C contents. Conclusion Obese rats induced by 36% fat-energy high-fat diet began to produce insulin resistance with about 50% of the obese rats at the end of the 8th week and the lipid metabolism were significantly disordered in insulin resistance rat.



Ginsenoside Rg1 Alleviated Epithelial Mesenchymal Transition of Airway in Chronic Obstructive Pulmonary Disease Rat

LIU Qian, GUAN Si-bin, HAN Feng-feng, GUO Xue-jun

2018, 38(6):  422-427.  DOI: 10.3969/j.issn.1674-5817.2018.06.004

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Objective To investigate the effect and the related mechanism of ginsenoside Rg1 in epithelial mesenchymal transition (EMT) in chronic obstructive pulmonary disease (COPD). Methods The COPD rat models were induced by exposure to cigarette smoke. Ginsenoside Rg1 was oral gavage dosed. The pulmonary function test and lung histologic HE staining were used to evaluate emphysema and small airway destruction. The expressions of E-cad and α-SMA were measured using immunohistochemistry assay, real-time PCR to evaluate the effect of ginsenoside Rg1 on EMT. Results Ginsenoside Rg1 ameliorated emphysema and airway destruction in COPD rats, improved pulmonary function, alleviated airway EMT. Conclusion Ginsenoside Rg1 can protest EMT in lung tissue,and may be act as potent intervention against small airway fibrosis in COPD.



Effects of Icariin on Human BALB/c-nu Prostate Cancer Model in Nude Mice

SONG Deng-peng, RAO Hong, HAN An-yan, WU Fu-yun, CHEN De-sen

2018, 38(6):  428-433.  DOI: 10.3969/j.issn.1674-5817.2018.06.005

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Objective To explore the possible mechanism of icariin inhibiting phosphoinositide 3-kinase/protein kinase B (P13K/Akt) signaling pathway and phosphorylation of related proteins in human prostate cancer (LNCaP) BALB/c-nu nude mice. Methods Totally 40 male BALB/c-nu nude mice were randomly divided into 4 groups (control group, prostate cancer group, positive drug group and experimental group) and the human prostate cancer model was established by intraprostate injection of LNCaP. The experimental group was injected 0.8% icariin by 0.1 mL/(10 g·d) tail vein, the positive group was injected 0.1 mL/10 g tail vein P13K/Akt inhibitor LY294002, and control group and the prostate cancer group were injected the same volume of normal saline. The body weight, wet weight, volume and P13K/Akt signaling pathway-related protein expression in nude mice were compared after 4 weeks of injection. Results The wet weight, volume, P13K, P-Akt, P-AR, AR-V7 and Calcitonin of prostate cancer tissues in the experimental group were lower than those in the model group (P<0.05), while E-cadherin was higher (P<0.05). Conclusion Icariin inhibits the progress of LNCaP by regulating PI3K/ Akt signaling pathway, which is related to the phosphorylation of Akt and AR.



Effects of Ganoderma Lucidum Polysaccharides on TLR4/NF-κB and Downstream Signal Transduction Pathway Activated by Lipopolysaccharide in Macrophage

YANG Yan-yan, XIE Jin-dong, ZHOU Jian-hua, LIN Wei, WANG Xun-Ii

2018, 38(6):  434-439.  DOI: 10.3969/j.issn.1674-5817.2018.06.006

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Objective To investigate the inhibitory effects of ganoderma lucidum polysaccharides (GLPs) on toll like receptor 4 (TLR4) signal transduction pathways and inflammatory cytokines regulating mechanism activateby lipopolysaccharide (LPS) in macrophage. Method Macrophage was stimulated with LPS for 16 hours and treated with the GLPs (low dose group 40 µg/mL, mid dose group 80 µg/mL, high dose group 160 µg/mL) for 2 hours simultaneously. In basis of this, the real-time fluorescent quantitative PCR and Western blotting methods were used to respectively detecting mRNA and protein of all groups of TLR4, myeloid differentiation factor 88 (MyD88), nuclear factor-kappa B (NF-κB), tumor necrosis factor receptor-associated factor 6 (TRAF-6), tumor necrosis factor alpha (TNF-α) and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1). Result (1) The mRNA relative expression were activated of TLR4 and primary element of downstream MyD88 dependent signal transduction pathway in LPS group, such as TLR4, MyD88, NF-κB and TRAF-6, which had significant differences compared with normal group (P<0.05). The GLPs could restrain in a degree the mRNA expression of TLR4, MyD88, NF-κB and TRAF-6 after being interfered in. The TLR4, MyD88 and TRAF-6 mRNA of GLPs-Mid and GLPs-High dose group had significant differences compared with LPS group (P<0.05). The NF-κB mRNA of all GLPs groups had significant differences compared with LPS group (P<0.05). And then, the relative expression of TLR4, MyD88, NF-κB and TRAF-6 protein in each group was consistent with the expression of mRNA. (2) Compared with normal group, the mRNA expression of TNF-α and LOX-1 were significantly increased in LPS group (P<0.05). The GLPs could restrain in a degree the mRNA expression of TNF-α and LOX-1 after being interfered in, which had significant differences compared with LPS group (P<0.05). And then, the expression of TNF-α and LOX-1 protein in each group was consistent with the expression of mRNA. Conclusion It was speculated that GLPs might restrain the expression of TLR4 and primary element of downstream MyD88 dependent signal transduction pathway. It was also illustrated that GLPs might restrain the expression of inflammatory cytokines such as TNF-α and LOX-1.



The Effect of Curcumin Pretreatment on Flora Expression of Ileum and Colon Contents in Heatstroke Rats with Simulated Desert Environment

XU Qin, DONG Xiang, LI Jian-ying, ZHANG Dong-hui, LI Jia-jia, YANG Ya-ping, YANG Qiu-yue, SHI Wen-hui, MA Na, SONG Lai-yang, LIU Jiang-wei

2018, 38(6):  440-445.  DOI: 10.3969/j.issn.1674-5817.2018.06.007

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Objective To investigate the effect of Curcumin on the flora expression of ileum and colon content in heatstroke rats with simulated dry heat desert environment. Methods Fifty male SD rats were randomly divided into 5 groups, with 10 rats in each group, including normal temperature and humidity group (control group), mild heat stroke group, moderate heat stroke group, severe heat stroke group, and severe heat stroke with Curcumin intervention group. The rats in Curcumin intervention group were intragastrically administrated with 200 mg/kg Curcumin suspension (1 mL/100 g body weight) for 7 successive days and the rest of the groups were continuously irrigated with saline for 7 days. Different degree of heat stroke rat models were established with the northwest special artificial environment laboratory chamber in our laboratory as previous ways. Animals were anesthetized and sterile specimens of ileum and colon contents were frozen in liquid nitrogen, PCR was used to analyze the expression of Escherichia coli, Enterococcus, Lactobacillus, Bacillus bifidus in the rats of each group. Results Heatstroke rats in simulated dry heat desert environment, the Escherichia coli and Enterococcus expression in their ileum and colon were increased significantly (P<0.05), the expression of Lactobacillus did not change significantly, and Bifidobacterium expression was significantly decreased (P<0.01). After intervention with Curcumin, Escherichia coli, Enterococcius expression was significantly decreased (P<0.05), the expression of Lactobacillus did not change significantly, but Bifidobacterium expression was significantly increased (P<0.01). Conclusion Curcumin may improve the expression of bacteria Bifidobacterium in the ileum and colon of heatstroke rats, inhibit opportunistic pathogenic Escherichia coli and enterococci proliferation in ileum and colon, therefore, keep the intestinal flora in homeostasis, reduce the degree of intestinal infection and enhance the rats' tolerance to heatstroke.

Probing on Establishment of Nail Regeneration Model in Mouse

ZHANG Xin-yan, GUAN Yu-ting, LI Da-li

2018, 38(6):  446-449.  DOI: 10.3969/j.issn.1674-5817.2018.06.008

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Objective To study the role of potential functional genes in the growth of mouse nails, and to establish and optimize mouse nail regeneration model of nail stem cells based on foreign research. Methods The 8-week-old C57BL/6 mice were selected as the experimental object, and precise position amputation surgery was performed after anesthesia under the microscope. The exploration and practice of a series of research methods such as animal modeling, real-time observation and recording, sampling measurement and section staining were carried out, and the research procedures were optimized as appropriate. Results A systematic study method for mouse nail regeneration animal model was developed. Conclusion This model can evaluate the growth of mouse nails at the animal level and tissue level, and it can be an effective means to study nail stem cells and related functional genes.



Study on Hematological and Biochemical Characteristics in Laboratory British-shorthair Cats

HU Ying, YANG Fei, CHEN Hong-ting

2018, 38(6):  450-454.  DOI: 10.3969/j.issn.1674-5817.2018.05.009

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Objective To study sexual and age differences of hematological and blood biochemical values in laboratory British-shorthair cats and to provide the basis for the cat related research. Methods Hematological parameters and blood biochemical indices of laboratory British-shorthair cats at 5 month old and 11 month old were measured, and the results were compared according to age and gender. Results There were age-related difference in 2 hematological indicators of female cats[hemoglobin(HGB), hematocrit(HCT)and in 5 indicators of male cats[HCT, mean corpuscular volume(MCV), erythrocyte(RBC), HGB, and mean corpsular hemoglobin(MCH)]. Sexual difference were observed in 2 hematological data [leukocyte(WBC), lymphocyte(LYMPH)from 5 month old cats and in 3 hematological data(RBC, HGB, LYMPH%) from 11 month old cats. Among the blood biochemical values, age-related difference exist in 4 parameters of female cats[alkaline phosphatase (ALKP), phosphorus(PHOS), cholesterol (CHOL) and creatinine(CREA)]as well as in 3 parameters of male cats(ALKP, PHOS, CREA), gender effect were also observed on CHOL of female cats and on 3 variables of male cats[albumin(ALB), CREA and alanine aminotransferase(ALT)]. Conclusion There are gender difference in some hematological and biochemical values of laboratory British-shorthair cat both in 5 month old and in 11 month old, as well as age-related difference between 5 month old cats and 11 month old cats. Our finding indicate that effects of gender and age should be taken into consideration in establishing hematological and biochemical reference intervals of laboratory British-shorthair cat.

The Literature Analysis on Dose of Intranasal Administration and Anesthetics in Mouse from China Journal Reports in Last 7 Years

LU Shuang-shuang, RONG Rong, SHI Xiao-meng, LIU Xiao-yu, LU Xuan-cheng

2018, 38(6):  455-458.  DOI: 10.3969/j.issn.1674-5817.2018.06.010

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Objective To investigate the domestic situation of intranasal administration in mouse. Method The data was retrieved from Wanfang database in the search term “mouse” and “intranasal administration” as defined by the 2011 to 2017 Journal Citation Reports. Perform a statistical analysis on the articles which meet the criterion. Result 770 articles were retrieved, and 513 articles described the dose of intranasal administration. In the 513 articles, there are 272 articles which the dose of intranasal administration is between 30 µL to 50 µL, and the ratio is 53.3%. The ratio of the articles that did not describe whether the personals use anesthetics or not in intranasal administration is 49.2%. The most used three anesthetics are diethyl ether (51%), barbiturates (24%) and chloral hydrate (12%) in the 249 articles which described the anesthetics used in intranasal administration. Conclusion Except the especial experiments, the dose of intranasal administration should be limited in 50 µL and the mouse should be anesthetized. The most used anesthetics in intranasal administration is diethyl ether. As diethyl ether is inflammable and explosive, we suggest it should be replaced by isoflurane in intranasal administration.

The Significance of Transgenic Animal Technology for Graduated Students in Medical University and Review of Teaching Experience

DU Xiao-yan, CHEN Bai-an, WU Yi, GUO Meng

2018, 38(6):  459-462.  DOI: 10.3969/j.issn.1674-5817.2018.06.011

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Transgenic animal is very important in medical and biological research fields. Many important investigation leaps in severe diseases are relied on transgenic animals. It is essential to offer a course of transgenic animal technology for graduated students. In present article, we shared a teaching review including teaching material compiling, the course outline, and test reformations to provide a reference and experiences for other colleagues.



Research Progress on Animal Model of Partial Bladder Outlet Obstruction

CHEN Lin, YANG Ya-fei, YANG Jin, HE Pin-lin, WANG Kai, WANG Yun-han, WANG Zi-li

2018, 38(6):  463-467.  DOI: 10.3969/j.issn.1674-5817.2018.06.012

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Constructing an ideal animal model of bladder outlet partial obstruction (pBOO) is the key to study the pathogenesis of bladder obstructive diseases and conduct drug therapy research. In this article, we summarized the animal model induction methods, including the characteristics and limitations of animal models induced by suprapubic bladder neck ligation, extraurethral splenic suture, and subcutaneous hyaluronate injection outside the urethra, and the characteristics and disadvantages of induced pBOO model.

Research Progress on Pathogenesis of Hereditary Diseases Caused by Mutations of Oxidoreductase DHTKD1 and Related Mouse Models

SHEN Yan, XU Wang-yang, ZHU Hou-bao

2018, 38(6):  468-472.  DOI: 10.3969/j.issn.1674-5817.2018.06.013

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As an oxidoreductase, dehydrogenase E1 and transketolase domain-containing 1(DHTKD1)catalyzes the oxidation dehydrogenation of 2-ketoadipic acid to produce glutaryl-CoA. Then the final product, acetyl-CoA enters the tricarboxylic acid cycle, finally producing adenosine-triphosphate (ATP) to provide energy for the cell and organism. So far, researchers have found that mutations in multiple loci of DHTKD1 gene are associated with mitochondrial dysfunction, leading to genetic diseases in human. This article focuses on the correlation between DHTKD1 gene mutation and human genetic diseases, and discussion about the corresponding mechanism. Furthermore, we dialectically discuss two available knockout mouse models.