Table of Content

25 August 2017, Volume 37 Issue 4

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Establishment and Characterization of Patient Derived Gastric Cancer Cell Lines

GE Xiao-mei, ZHANG Yi-xin, XIE Fu-bo, LIU Ji-bin, YANG Lei, QU Ying-ying, GU Ying, Li Xue-ting, YANG Wei-min, LIU Xi-peng, ZHOU He, QIANG Fu-lin

2017, 37(4):  257-265.  DOI: 10.3969/j.issn.1674-5817.2017.04.001

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Objective To establish and characterize primary gastric cancer cell lines derived from gastric cancer patient-derived xenograft (PDX) models. Methods Tumors from 12 stable gastric cancer PDX models were selected to establish primary tumor cell lines when the tumor volume reached 500~900 mm³. The established cell lines were characterized by cell morphology, chromosome, STR and immunohistochemical analyses, in vivo tumorigenicity, as well as drug sensitivity in vitro and in vivo. Results Six primary gastric tumor cell lines were successfully established. GAXC031 and GAXC066 were selected and characterized thoroughly. STR analysis confirmed that these 2 cell lines were consistent with their parental PDX models. These 2 cell lines presented the morphological, histological and genomic characteristics of malignant gastric cancer cells. They generated xenograft readily in nude mice and these tumors maintained histology characteristics of the parental PDX tumors. Moreover, compared to the parental PDX models, these 2 cell lines initiated xenografts with higher take rate, shorter incubation period, and better uniformity in tumor growth. In vitro and in vivo drug sensitivity test showed that GAXC031 was more sensitive to 5-fluorouracil than that of GAXC066, while both cell lines were sensitive to cisplatin. Conclusions Six patient derived gastric cell lines were successfully established from PDX models in this study. Deep characterization showed that these cell lines maintained many characteristics of the original PDX models with improved take rate, latent period and uniformity, therefore represent useful tools in basic research and drug screening.



An Experimental Study on Repairing Femoral Condyle Defect by Printing Porous Titanium with 3D Technology in Rabbits

DENG Wei, ZHENG Xin, SHEN Ye-shuai, RUI Min, WANG Fan, LI Sai, GUO Ji-qiang, LIU Hong, GUO Kai-jin

2017, 37(4):  266-272.  DOI: 10.3969/j.issn.1674-5817.2017.04.002

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Objective To investigate the ability of the 3D printed porous titanium material for bone defect repairing and bone formation. Methods The critical bone defect models (6 mm in depth and 10 mm in diameter) were made of 20 six-month-old New Zealand white rabbits’ femoral condyle. One of the porous titanium material was swiftly implanted into the bone defect area (experimental group, n=20), with the opposite side as control group (also the blank group, n=20), washed with saline and sutured without any implants. The samples were harvested and observed by X-ray and CT examinations at 3 days and 4, 8, 12 weeks after the operation to assess bone growth in the bone defect area. At 12 weeks after the operation, all rabbits were euthanized and evaluate the bone defect repair through X-ray, CT, histology and Micro-CT. Results Both the experimental group and control group recovered with good condition. At 12 weeks after the operation, we can see that the bone defect area was filled with new bone in the experimental group after removing the soft tissue surrounding the bone defect area; In contrast, the bone defect in control group was seen hollow and covered with fibrous capsule. According to the radiological observation, the demarcation line between the defect area and the surrounding area was obscure in the experimental group; the bone defect area in control group saw an entire translucency with no obvious shadow reflecting the new-borne bone. The Micro-CT observations proved fresh bone tissue formed in the plantation area at 12 weeks after the operation, and the surrounding area could see bone trabecula grown, In comparison, there was no obvious bone formed in the defect area of control group, which was filled by fiber tissue and relatively less bone trabecula and new-borne bone tissue. The results of un-decalcificated bones slicing showed the implanted material appeared combined with osteoblast, and the mature harversian system was seen distributed within the new-borne bone; for control group, a small amount of unordered bone formed in the defect area, filled with a lot of fiber texture. The area ratio of new-borne bone and the defect area was calculated via the IPP5.1 software, producing differences that have statistical significance. Conclusion A series of experiments prove that the porous titanium material has high biosecurity and ability to promote bone growth and healing. The new-borne bone tissue can grow and fill the lacuna, thereby effectively repairing bone defect. In short, the 3D printed porous titanium is a kind of promising repairing material in the tissue engineering field.



Application of Digital Three-dimensional Reconstruction Technique in Anatomical Measurement and Investigation of Radius in New Zealand White Rabbits

RUI Min, ZHENG Xin, LI Cheng-yu, JIANG Wei, YAO Sheng-cheng, GUO Kai-jin

2017, 37(4):  273-277.  DOI: 10.3969/j.issn.1674-5817.2017.04.003

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Objective To obtain anatomical data of radius in New Zealand white rabbits with different age by three-dimensional reconstruction technique of CT images and provide references for rabbit bone defect experiment and bone tissue engineering scaffold manufacture. Methods Forty male New Zealand white rabbits aged 3 months and 6 months were included respectively in this study. According to their age, the rabbits were divided into group A (3 month) and group B (6 month). Computed tomography(CT) scanning of bilateral radius was conducted and the contours of 3D-reconstructed radius were observed by Mimics l0.0, the anatomical parameters including the length of radius, transverse diameters and sagittal diameters of middle of radius, front curvature angleandlateral curvature angle were measured respectively. Results The contours of radius were bulky at ends, narrow in the middle. The obvious camber was observed in radial middle, and the mean front curvature were (24.19±1.18)° in group A and (23.97±1.35)°in group B, and the lateral curvature angles were (26.13±1.04)°in group A and (25.66±1.23)°in group B respectively, and there was no significant difference between the two groups (P>0.05). The straight and curvilinear length of radius were (63.05±1.33) mm and (65.03±1.10) mm in group A, and (66.56±1.53) mm and (70.41±1.06) mm in group B respectively, with no significant difference (P<0.05). The transverse diameters of the middle of radius were (4.24±0.19) mm in group A and (4.30±0.23) mm in group B. The sagittal diameters of the middle of radius were (3.23±0.16) mm in group A and (3.29±0.18) mm in group B respectively. There was no significant difference between the two groups with regard to sagittal diameters (P>0.05). And the transverse diameters and the sagittal diameters of the middle of radius were not correlated with length of radius (P>0.05). Conclusions It is feasible to accurately identify the contours and bony landmarks on the three-dimensional reconstruction of the radius. Digital measurement can provide accurate parameters for establishment of bone defect model and manufacture of bone-tissue engineering scaffold.



Immunologic Mechanism of Inhibition on Gastrointestinal Tract Tumors of ZHEMU Extract

XIE Jia-jun, ZHANG Guo-ming, QIAO Zheng-dong, LU Hong-qi, YAN Hui-fang, CHENG Mao

2017, 37(4):  278-282.  DOI: 10.3969/j.issn.1674-5817.2017.04.004

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Objective To observation the changes of immune function in normal or tumor bearing animal after dose ZHEMU extract. Methods The carbon particle clearance test, macrophage phagocytosis test. NK cells activity and lymphocyte transformation in tumor bear animal are detected. Results The carbon granula clearance ability and phagocytosis of macrophage cell in abdominal on normal mice, and the lymphocyte transformation and natural killer (NK) cell activity in tumor bearing mice were enhanced after dosing, and the lowest effective dose were 0.3 g/kg, equivalent to the crude herb 5.46 g/kg, which was 1.3 times of clinical daily dose. Conclusions ZHEMU extract can restrain the growth of gastrointestinal tract tumors, and its mechanism may be related to the direct inhibition of the gastrointestinal tract cancerous cells growth and the improvement of the body's specific and non-specific immune function.

The Feasibility of Social Housing for Adult Laboratory Rabbits

LI Cai-yun, JIANG Zi-rui, PAN Xue-ying, SONG Zheng, CHANG Yan

2017, 37(4):  283-287.  DOI: 10.3969/j.issn.1674-5817.2017.04.005

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Objective To study the feasibility of social housing for adult laboratory rabbits. Methods Totally 38 New Zealand white rabbits aged at 6 months with 19 males and 19 females were single housed. Food intake and body weight were observed one day prior to social housing, then the rabbits were divided into 2 groups (19 male, 19 female), each group respectively housed in 8 cages (5 cages with 2 rabbits, 3 cages with 3 rabbits) for 7 days, food intake, body weight, fighting and injury were observed during 7 days social housing, and then single housed for 21 days. Results The food intake of female rabbits was decreased compared with that of being single housed, the normal growth of body weight was affected; high fighting frequency was observed at the first day of social housing, fighting frequency reduced or stopped after social hierarchy established; no obvious injuries were observed. The amount of food intake and body weight were reduced significantly for adult male rabbits during social housing, frequent and continuous fights were observed that caused serious injuries even death. Conclusions Adult female rabbits can be social housed under proper nursing by animal care staff during the initial stage of social housing, social housing for adult male rabbits is infeasible.

Effect of Sodium Pantothenate on Cardiovascular in Beagle Dogs

ZHANG Cheng-mei, JI Jian-bo, WU Chuan-tao, SONG Wen-yan, SHI Yan-qiu

2017, 37(4):  288-291.  DOI: 10.3969/j.issn.1674-5817.2017.04.006

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Objective To observe the general influence of sodium pantothenate on the cardiovascular indicators of Beagle dogs. Method Sixteen healthy Beagle dog with half female and male. They were randomly divided into four groups: high, medium and lower dose of sodium pantothenate with 0.26 mg/kg, 0.52 mg/kg, 1.04 mg/kg and normal saline control group (NS). The dogs were anesthetized with intravenous injection of 3% pentobarbital sodium. The unilateralcarotid artery was dissociated so as to insert the artery cannulae for measuring systolic and diastolic blood pressure. Then unilateralfemoral vein is isolated for a single intravenous injection of 1 mL/kg sodium pantothenate, and electrodes were placed into their limbs to record the indexs of blood pressure, heart rate and electrocardiograph before administration and at different time points after drug administration while each observation index of the Beagle dogs was constant. Result In this experiment, sodium pantothenate did not evidently change the value of blood pressure, heart rate and electrocardiographic in Beagle dogs after intravenous injection (P>0.05). Conclusion The sodium pantothenate has no evident effects on the cardiovascular indicators of Beagle dogs.

Influence of Butyric Acid Bacteria on Harmful Gas Emissions in Mice

PAN Li-jie, YU Wen-lan, REN Bin-bin, CHEN Jia, MA Xiao-li, ZHANG Jun-jun, LIU Yong-lun, LIU Zhong-hua

2017, 37(4):  292-294.  DOI: 10.3969/j.issn.1674-5817.2017.04.007

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Objective To explore the influence of butyric acid bacteria on harmful gas emissions in KM mice. Methods Eighty KM mice were randomly divided into four groups (group I, II, III and IV) with equal number of females and males in each group. The mice in group II, III and IV were fed with butyric acid bacteria at the dose of 500 mg/kg、1000 mg/kg、1500 mg/kg by gavage for 21 d, respectively, and group I as control. The concentration of ammonia and hydrogen sulfide were monitored every day, and the growth performance, the activity of urease in faeces in each group were tested at 21 d. Result Compared to the control group, the average daily weight gain of group III and group IV were improved. On day 7 of gavage, the ammonia emission of group III and IV were significantly lower than that of group I and group II (P<0.05), while the H₂S emission were not edfected. The urease activities of group III and IV were lower than group I and II, but not significantly (P>0.05). Conclusion The butyric acid bacteria may improve the production performance and reduce the ammonia emission of mice.



An Introduction of 《Public Health Service Policy on Humane Care and Use of Laboratory Animals (Revised 2015)》

SHENG Jun-jie, HU Qing-yuan, WANG Ke-zhou, LV Su-jun

2017, 37(4):  303-308.  DOI: 10.3969/j.issn.1674-5817.2017.04.010

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《Public Health Service Policy on Humane Care and Use of Laboratory Animals》is an important part of the legal system of laboratory animal welfare in the United States. It is the main policy for U.S. government to implement the management of laboratory animal industry, and also the foundation for caring and using laboratory animals in scientific research. The PHS Policy covers all the research activities conducted or supported by the Public Health Service (PHS). The Office of Laboratory Animal Welfare, National Institute of Health (NIH), which has responsibility for the general administration and coordination of the Policy, revised the PHS Policy in June 2015. The authors translated the full-text PHS Policy into Chinese and provided reference to understand and study the laws and regulations on laboratory animal welfare of U.S. The essay may be used for reference to learn the methods and experience on humane care and use of laboratory animals.

Effects of Irx5a Gene Overexpression on Early Hematopoietic in Zebrafish Embryos

JIN Lu, LI Yan, LI Zhi-cao, WU Xi-jun, ZHOU Yan-hua, HE Zhi-xu, SHU Li-ping

2017, 37(4):  309-314.  DOI: 10.3969/j.issn.1674-5817.2017.04.011

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Objective To investigate the expression and influence of early hematopoietic related genes in zebrafish embryos. Method The single-cell-stage zebrafish embryos were micro-injected with iroquois homebox 5a-enhanced green fluorescent protein (Irx5a-EGFP) mRNA as experimental group, EGFP mRNA as experimental control group, wild type zebrafish as blank control group. Comparised the hematopoietic transcript factor genes with three groups by quantificational reverse transcription-PCR (qPCR). Such as primitive hematopoiesis scl and lmo2, definitive hematopoiesis c-myb and runx1, erythroid hematopoiesis gata-1, myeloid hematopoiesis pu.1. The zebrafish embryos’ time point are 9 h、12 h、18 h、24 h、30 h post fertilization (pf). Results From the real-time fluorescence quantitative PCR can be seen in the original hematopoietic transcription factor lmo2 and scl, the experimental group from 9 hpf to 24 hpf significantly lower than the other two groups. In the targeted hematopoietic transcription factor, c-myb experimental group from 12 hpf to 24 hpf significantly lower than the other two groups, runx1 experimental group from 9 hpf to 30 hpf significantly lower than the other two groups. Myeloid hematopoietic transcription factor pu.1 and erythroid hematopoietic transcription factor gata-1, the experimental group from 9 hpf to 30 hpf were significantly lower than the other two groups. The differences were statistically significant. Conclusion Irx5a gene plays an inhibitory role in the transcriptional regulators of primitive hematopoiesis, adult hematopoiesis, erythroid hematopoiesis and myeloid hematopoiesis in the zebrafish.

Effects of Cigarette Smoke on Lung Function and Serum Cytokine Levels in Chronic Obstructive Pulmonary Disease Rat

XIONG Ying, ZHANG Jin-juan, LI Lin, CHEN Yan, WU Yan-jun, HUANG Hui-yan

2017, 37(4):  315-319.  DOI: 10.3969/j.issn.1674-5817.2017.04.012

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Objective To investigate the influence of cigarette smoke on lung function and serum cytokine levels in chronic obstructive pulmonary disease (COPD) rats which were established by the complex stimulation of lipopolysaccharide (LPS) and smoked cigarettes. Methods Seventy-two male SD rats were randomly divided into six groups, 4 weeks, 6 weeks and 8 weeks model group, 3 weeks and 6 weeks natural healing group, and control group. A COPD rat model was established by the complex stimulation of two times LPS injection and smoked cigarettes. The lung function of the rats was measured during passive smoking and quit smoking. At the same time serum interleukin-8(IL-8), IL-10 and tumour necrosis factor-alpha (TNF-〈) levels were monitored by Enzyme-linked immunosorbent assay. Correlation analysis concerning cytokine levels and pulmonary function index were conducted. Results Compared with control group, forced expiratory volume in 0.3 second (FEV_0.3), forced expiratory volume in 0.3 second as a percentage of forced vital capacity (FEV_0.3/FVC) and IL-10 were decreased (P<0.05), IL-8 and TNF-〈 levels were increased in the model groups (P<0.05). And as the building time prolonged, the changes of those cytokines and lung function indexes were more obvious than before. IL-8 and TNF-〈 levels were inversely associated with FEV_0.3/FVC(P<0.001, r=-0.579; P=0.034, r=-0.354). Compared with control group, IL-8 and TNF-〈 levels were decreased in the 6 weeks healing group, at the same time IL-10 levels was increased(P<0.05), the FEV_0.3 and FEV_0.3/FVC were decreased in the healing groups (P<0.05). Conclusions Cigarette smoke is the primary cause of changes on cytokine levels in this animal model.



The Protective Effect of Oxymatrine on Asthma Mice

LAN Lu-sha, ZHAO Bing-bing, YANG Hong-yu, LUO Hong, JIANG Yan

2017, 37(4):  320-323.  DOI: 10.3969/j.issn.1674-5817.2017.04.013

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Objective To study the protective effect of Oxymatrine (OMT) on asthma mice. Methods The asthmatic BALB/c mice were induced by ovalbumin (OVA) and aluminium hydroxide.The general situations and weights of mice were observed every day. Then these mice were treated by using OMT and/or dexamethasone and were sacrificed at 5 or 15 days after treatment. The bronchoalveolar lavage fluid (BALF) and lung tissue of mice were obtained. Their pathological reaction was observed through HE stain. The total number of cells and eosinophilscells in BALF were counted. Result OMT can improve the weights of asthma mice. On day 5 and 15, the pathological reactions of the lung tissue in each treatment groups were the same with that of the asthma group, but the number of inflammatory cells in lung tissue was much less than that of the asthma group. The total number of cells and eosinophils cells in BALF were reduced compared with the asthma group (P<0.05 or P<0.01). Conclusion OMT has protective effect on the asthma mice.



Primiparous Reproductive Performance of Outbred Guizhou Miniature Pigs and its Correlation and Path Analysis

CHEN Ming-fei, YAO Gang, ZHAO Hai, QIAN Ning, WU Shu-guang

2017, 37(4):  324-327.  DOI: 10.3969/j.issn.1674-5817.2017.04.014

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Objective To screen the reproductive traits significantly associated with reproductive performance of outbred Guizhou Miniature Pigs. Methods Ten reproductive traits of 30 gilts were determined and analyzed with correlation and path analysis.The traits were as following: totol number of born (X1), number of alive born (X2), average weight at birth (X3) and litter weight (X4), average number alive(X6) and weight (X5) and litter weight (X7) at 21 days, number weaned (X8), average weaning weight (X9) and weaning weight of litter (Y). Result There were extremely significant (P<0.01) positive correlation between weaning weight of litter and other reproductive traits. Path analysis showed that X7~X9 had positive direct effects on weaning weight of litter but X5~X6 had positive indirect effects, and X1~X4 had weak effects. Conclusion Compared with genetic factor, the life environment had decisive influence on weaning weight of litter.