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June 15,2013
Volume 6,Issue 11
Pages -
Subject Area:Animal Nutrition,Animal Breeding,Health Administration,Environmental Hygiene,Epidemiology

Title: Eukaryotic expression and biological activity of avian β-defensin 6 and its derivative

Authors: CAO Yanping, MA Qingquan, SHAN Anshan
PP: 1070-1077
Abstract: To explore the effect of disulfide bonds on antimicrobial activity of avian β-defensin and to obtain the eukaryotic expression of avian β-defensin 6 (AvBD-6), all the Cys residues in the maturation region of AvBD-6 were removed and then obtained the derivative AvBD-6-B. According to the codon bias of Picha pastoris, the genes of AvBD-6 and AvBD-6-B were designed and synthesized. Then, the recombinant plasmid was constructed through inserting AvBD-6 and AvBD-6-B genes into the restriction sites between XhoⅠ and EcoRⅠ of the eukaryotic expression vector pGAPHαM. The Bgl Ⅱ-linearized recombinant plasmids pGAPHαM-AvBD-6 and pGAPHαM-AvBD-6-B were transformed into P. pastoris GS115 by electroporation. The recombinant AvBD-6 and AvBD-6-B were expressed in 100 mL yeast extract peptone dextrose (YPD) (including 1% casein hydrolyzate). Tricine-SDS-PAGE demonstrated that both AvBD-6 and AvBD-6-B were expressed in P. pastoris, the concentration of recombinant AvBD-6 and AvBD-6-B reached 114.9 mg/L and 93.8 mg/L, respectively. The bacteriostatic experimental results showed that this two expression products exhibited the same antimicrobial activity against Bacillus subtilis CMCC 63501. However, for Staphylococcus aureus ATCC29213, the antimicrobial activity of AvBD-6-B was higher than AvBD-6, indicating that the disulfide bonds in AvBD-6 were not necessary for antimicrobial activity. This research provides a theoretical basis for achieving industrial production of antimicrobial peptides using genetic engineering technology.
Keywords: animal nutrition; avian β-defensin 6; disulfide bonds; Pichia pastori; expression

Title: In situ hybridization protocol using a new type of nucleic acid probe based on quantum dots

Authors: HE Jiawei, ZOU Qian, LIU Jun’an, WANG Xu, CAO Jianhua, ZHAO Shuhong
PP: 1064-1069
Abstract: Quantum dot (QD) is a new material in biomarkers, which has significant advantages of stable fluorescence, high sensitivity, wide fluorescence emission spectrum and harmonious biocompatibility. In this study, the cores of CdSe/ZnS-QDs was chemically synthesized, and then they were covalently binded with nucleic acid probes from glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene to form QD-DNA probes. This probe was proved to have the characteristics of QD and nucleic acid probe by electrophoresis and fluorescence detection. Furthermore, the QD-DNA probes were used in the in situ hybridization to conduct for the target gene detection in longissimus dorsi muscle in Yorkshire pigs. The results showed that QD-labeled probe had distinct positive signals. These results provided an experimental foundation for in situ hybridization based on QD-labeled probes, and shed light on the biomaterial applications of QDs. In addition, this research also provided a feasible method for the application of in situ hybridization in animal breeding.
Keywords: animal breeding; quantum dot; glyceraldehyde-3-phosphate dehydrogenase gene; DNA probe; in situ hybridization

Title: Prokaryotic expression and polyclonal antibody preparation of VP1 gene of duck hepatitis virus type 1 E53

Authors: FU Peifen, LIU Yan, LI Xin, MU Xiaoyu, DONG Jingquan, WANG Junwei, MA Bo
PP: 1058-1063
Abstract: According to the genome sequence of duck hepatitis virus type 1 (DHV-1) E53, VP1 gene was amplified by RT-PCR and cloned into prokaryotic expression vector pET-30a to construct pET-30a-VP1 vector. After sequencing, the recombniant expression vector pET-30a-VP1 was transformed into the host Rosetta(DE53)plysS. SDS-PAGE analysis indicated that the VP1 fusion protein was expressed in Escherichia coli successfully after induced by isopropyl β-D-1-ihiogalactopyranoside (IPTG). Western blotting analysis showed that the fusion protein was expressed correctly. Then the rabbits were immunized with purified recombinant VP1 protein and Western blotting analysis showed that the sera from immunized rabbits had specific reaction with DHV-1. In addition, the antibody titer was 1:12 800 by indirect ELISA, and neutralization test showed that its neutralization antibody titer was up to 1:32. Therefore, the expressed recombinant protein and polyclonal antibodies from this study provided important data and material for the further study of VP1 protein function and the development of diagnostics.
Keywords: veterinary immunology; duck hepatitis virus type 1; VP1; prokaryotic expression; immunogenicity; neutralization

Title: Effect of fixed rotational grazing time on spatial heterogeneity of soil available nitrogen in Stipa breviflora desert steppe

Authors: WEI Zhijun, LIU Hongmei, LV Shijie, WU Yanling, SUN Shuqing, YUN Xiangjun
PP: 1050-1057
Abstract: This study focused on spatial heterogeneity distribution of soil available nitrogen in fixed rotational grazing time. And the effect of spatial distribution of soil available nitrogen on the response of fixed rotational grazing time in desert steppe was investigated using variation analysis, geostatistical analysis and fractal theory methods. The results showed that the soil available nitrogen content significantly increased with rotational grazing time delaying, and the spatial distribution differences decreased both in unit mean and spatial autocorrelation, while the fractal dimension increased gradually. Furthermore, in the mid-rotational grazing areas (RG4), the spatial distribution of soil available nitrogen had special distribution characteristics, which may had higher complexity spatial distribution. Therefore, it was not appropriate for grazing with fixed rotational grazing time in rotational grazing grassland.
Keywords: grassland science; fixed rotational grazing time; desert steppe; spatial heterogeneity; soil available nitrogen

Title: EGFR activation is involved in TRAIL resistance in gastric cancer SGC7901 cells

Authors: XU Ling, QU Xiujuan, LIU Yunpeng, HOU Kezuo, ZHANG Ye, LIU Jing
PP: 1046-1049
Abstract: The resistant mechanism of gastric cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is still unclear. The research is to identify whether the activation of epidermal growth factor receptor (EGFR) is involved in TRAIL resistance in gastric cancer SGC7901 cells. In this study, protein expression was detected by Western blotting and cell apoptosis was determined using flow cytometry. Transient transfection with small interfering RNA (siRNA) was used to inhibit the expression of EGFR. The results showed that treatment with TRAIL induced the phosphorylation of EGFR in SGC7901 cells. After SGC7901 cells were transiently transfected with EGFR siRNA for 48 h, the expression of EGFR was obviously decreased. Compared to non-silencing control, TRAIL-induced EGFR phosphorylation was downregulated in EGFR knockdown clones. The inhibition of EGFR expression finally increased TRAIL-induced apoptosis in SGC7901 cells (P<0.05). Therefore, the activation of EGFR was one of the reasons of TRAIL resistance in gastric cancer SGC7901 cells.
Keywords: oncology; gastric cancer; tumor necrosis factor-related apoptosis-inducing ligand; epidermal growth factor receptor; resistance

Title: Study on osteogenic differentiation potential of umbilical cord mesenchymal stem cells

Authors: YU Yongchun, WANG Weixin, SUN Jie, LIU Lei
PP: 1040-1045
Abstract: Objective: To explore the osteogenic differentiation potential of umbilical cord mesenchymal stem cells and provide a rich source of stem cells for bone tissue engineering. Methods: Trypsin and collagenaseⅠdigestion method was used to obtain the umbilical cord mesenchymal stem cells. Surface markers were identified using immunocytochemical method. Chemical induction was used to induce umbilical cord mesenchymal stem cells into osteoblasts. Inverted microscope was used to observe the morphological changes. Alizarin red staining was used to stain the induced cells to check the osteogenic differentiation. Results: The cells isolated from umbilical cord using trypsin and collagenaseⅠdigestion method showed the adherent growth and fibroblast morphology. Immunocytochemistry showed that the umbilical cord mesenchymal stem cells were positive for CD29, 40 CD44 and CD105 while negative for CD31, CD34 and CD105. Umbilical cord mesenchymal stem cells were accumulated into growth after osteogenic induction and the nodular clumps formed in the center of the gathering cells. Then the osteogenic differentiation was determined by the production of calcium deposite detected by alizarin red staining. Conclusion: The umbilical cord mesenchymal stem cells have the osteogenic potential, which can provide the seeding cells with an abundant source, low immunity, and poor differentiation for bone tissue engineering.
Keywords: oral and maxillofacial surgery; osteogenic differentiation; chemical induction; umbilical cord mesenchymal stem cells

Title: miR-133a increases the sensitivity of laryngeal cancer cells to cisplatin and affects expression of ATP7B

Authors: WANG Xurui, WANG Ping, ZHAO Xiaodong, DAI Xiaotian, ZHU Wei
PP: 1033-1039
Abstract: Objective: To investigate the influence of miR-133a on chemosensitivity of laryngeal carcinoma drug resistant cells, and clarify the molecular mechanisms of miR-133a regulation of cisplatin on killing tumor cells. Methods: miRNA were firstly poly tailed, and then applied PCR to detect miR-133a levels in normal pharyngeal epithelial cells NP69, Hep-2 and Hep-2/VCR. Then, lentiviral recombinant vector pEZX-miR-133a was transfected into Hep-2 cells and Hep-2/VCR cells, the effects of different concentrations of cisplatin on cell survival were observed by using cell viability experiments. The differences of ATP7B expression at transcription and protein level after being transfected pEZX-miR-133a were also detected by immunofluorescence staining and real-time PCR. Results: ATP7B fluorescence value was significantly higher in Hep-2 and Hep-2/VCR than in the NP-69 cells. The survival of pEZX-miR-133a Hep-2/VCR cells was significantly lower than the pEZX-control Hep-2/VCR cells after adding 50 μmol/L and 100 μmol/L cisplatin. In addition, miR-133a reduced transcription and expression of ATP7B in Hep-2/VCR cells. Conclusion: miR-133a could increase the chemosensitivity of laryngeal carcinoma cells to cisplatin, and the mechanism might be related to down-regulating of ATP7B protein expression by miR-133a.
Keywords: otorhinolaryngology; laryngeal carcinoma; miRNA; chemotherapy; ATP7B

Title: TAK-778 induces osteogenesis in ovariectomized rats through an estrogen receptor-dependent pathway

Authors: LIU Pengcheng, CAI Ming
PP: 1027-1032
Abstract: Objective: To test if TAK-778 induces osteogenesis through an estrogen receptor (ER) dependent pathway using an ovariectomized (OVX) rat model. Methods: Two weeks after test animals underwent ovariectomy, TAK-778 and/or tamoxifen was administered orally over 3 months. Vehicle-treated and sham-operated rats were served as controls. The bone mineral density (BMD) of the lumbar vertebrae and sagittal two-dimensional images of the L3 vertebral body were measured. In addition, bone formation rates (BFR) and serum calcium and osteocalcin levels were investigated. Results: TAK-778 significantly increased BMD, serum calcium and osteocalcin levels, and BFR when comparing to those of the vehicle-treated group. However, tamoxifen clearly inhibited the increase in these parameters induced by TAK-778. In addition, micro-CT tomography scanning showed that treatment with TAK-778 increased the structure model index (SMI), bone volume/tissue volume (BV/TV), and trabecular thickness (Tb.Th) parameters, while decreased the trabecular spacing (Tb. Sp) in OVX rats. Tamoxifen suppressed these effects when was combined with TAK-778. Conclusion: Taken together, the present study showed that TAK-778 enhanced osteogenesis in ovariectomized rats through an estrogen receptor-dependent pathway.
Keywords: surgery; osteogenesis; TAK-778; tamoxifen; estrogen receptor; ovariectomized rats

Title: Study on the internal control factors of new rural cooperative medical system based on the internal control theory

Authors: CHEN Li, FENG Zhanchun
PP: 1021-1026
Abstract: For the fund management of new rural cooperative medical system, the problems of the fund security and its irrational use exist. This article mainly applied the internal control theory to analyze the new rural cooperative medical care fund management, and found that the internal control of new rural cooperative medical system included 5 aspects: environment controlling, risk assessment, activity controlling, information and communication, monitoring and evaluation. According to the above 5 aspects, we evaluated and controlled the new rural cooperative medical operating effect, which had a great significance in securing the safety of the new rural cooperative medical fund, guaranteeing the fund utilized reasonably and improving the benefiting degree of farmers who participated in the new rural cooperative medical system.
Keywords: health management; new rural cooperative medical system; internal control theory; fund management

Title: Sequence analysis on complete genome of an Enterovirus 71 strain SDLY96 isolated from a severe case

Authors: SI Luying, WEN Hongling, GAO Feng, ZHAO Li, SONG Yanyan, XU Hongzhi, YUAN Xiaojing, WANG Zhiyu
PP: 1014-1020
Abstract: Objective: To analyze the complete genome sequence of an Enterovirus 71 (EV71) strain SDLY96 isolated from a severe case and lay the foundation for constructing infectious clone of EV71. Methods: The complete genomes of SDLY96 from a sufferer of hand-foot-and-mouth disease (HFMD) in Linyi city of Shandong province were amplified by RT-PCR. The homology of nucleotide and amino acid sequences were aligned, and phylogenetic relations of the sequences were analyzed. Results: The full length of SDLY96 strains was 7 405 bp. SDLY96 strain shared the high homology with Fuyang strain EU703813 both in nucleotide and in amino acid sequences, with 98.4% identity in nucleotides and 99.1% in amino acids. The full VP1 region based phylogenetic analysis also indicated that strain SDLY96 had the close genetic relationship with Fuyang strain EU703813. Conclusion: SDLY96 should be classified as C4a branch of subgenogroup C4 of EV71, and SDLY96 strain has close genetic relationship with Fuyang strain.
Keywords: sanitary technology; Enterovirus 71; sequence analysis; complete genome

Title: Dose evaluations for the examinees undergoing common types of diagnostic X-ray projections

Authors: LIU Haikuan, ZHUO Weihai
PP: 1008-1013
Abstract: Objective: To evaluate the entrance surface doses (ESD), organ doses and effective doses from common projections of diagnostic X-ray examinations. Methods: Based on the in-phantom measurements, the ESD and organ doses of 6 common projections were measured with the radiophotoluminescent glass dosemeters, and the effective doses were evaluated by combining the organ doses with the corresponding tissue weighting factors. Results: The ESD, organ doses and effective doses varied with 6 kinds of projections, and the mean ESD ranged from 0.25 to 8.64 mGy. In addition, the values and distributions of organ doses differed from different projections, and the mean effective doses ranged from 0.04 to 1.08 mSv among 6 projections. Conclusion: The doses for examinees undergoing different projections of diagnostic X-ray examinations are different, the effective doses can be fast estimated with the ESD and the conversion coefficients for different projections.
Keywords: radiohygiene; entrance suface dose; antropomorphic phantom; radiophotoluminescent glass dosemeters; organ dose; effective dose

Title: Effects of iodine-deficient on apoptosis in hippocampus of pups

Authors: DONG Jing, WANG Yuan, MIN Hui
PP: 1004-1007
Abstract: Objective: To observe the effects of iodine deficiency on apoptosis in hippocampus of pups. Methods: Two-month old heathy female Wistar rats (n=14) after pregnancy were randomly divided into control group and iodine-deficient group (7 in each group). From gestational day 6 to postnatal day (PND) 28, iodine-deficient group was administered with iodine-deficient diet and tap water, while control group was fed with normal diet. 5 pups from each group were sacrificed and intracardiac perfused in PND14, PND21 and PND28 respectively. Brains were removed，fixed and sectioned coronally. The apoptosis in the hippocampus was determined by TUNEL assay. Results: In PND14 and PND21, the apoptosis cells was increased in the iodine-deficient group in hippocampus (P<0.05). In PND28, the positive cells were scarcely observed in all groups. Conclusion: Iodine deficiency may result in the increase of apoptosis cells and delay of apoptosis peak in hippocampus of pups.
Keywords: environmental hygiene; iodine deficiency; hippocampus; apoptosis

Title: Relationship between FABP2 Ala54Thr polymorphism and the risk of colorectal cancer

Authors: XU Lin, FENG Fei, HU Xiaoqin, YAN Jin, YANG Yanfang
PP: 997-1003
Abstract: Objective: To explore the relationship between the human intestinal fatty acid binding protein 2 (FABP2) Ala54Thr polymorphism as well as gene-environment interaction and colorectal cancer (CRC). Methods: Using a 1:1 matched case-control study to collect the general demography data and eating habits of 151 new cases of CRC and 151 healthy subjects (control) by face-to-face interview. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect the FABP2 Ala54Thr genotypes. Conditional logistic regression was used to analyze the relationship between FABP2 Ala54Thr polymorphisms and environmental factors with CRC. Results: There was no statistically significant in the distribution of alleles frequency and genotype frequency between CRC group and control group (P>0.05). However, there were positive interaction between polymorphism of FABP2 Ala54Thr and consumption of pickled vegetables［odds ratio, OR=4.83, 95% confidence interval (CI): 1.23-18.89］ and sweet food (OR=2.60, 95%CI: 1.15-5.88). Conclusion: No relationship is found between FABP2 Ala54Thr polymorphisms and CRC. However, the interaction between FABP2 Ala/Thr, Thr/Thr genotype and environmental factors may increase the risk of CRC.
Keywords: epidemiology; colorectal cancer; genetic polymorphism; fatty acid binding protein 2; environmental factor; interaction

Title: Prophylactic effect of capsaicin on alimentary obesity of rats

Authors: QIN Hong, YUAN Quan, REN Guofeng
PP: 992-996
Abstract: Objective: To observe the effects of capsaicin (CAP) on body fat, plasma lipids and glucose in overweight rats and investigate the prophylactic effect of CAP on alimentary obesity. Methods: 40 rats were divided randomly into five groups: normal-control group (ND), model group (HD), low CAP supplement group (HD+5), middle CAP supplement group (HD+10) and high CAP supplement group (HD+20). The rats were fed with high fat and carbohydrate diet to induce overweight, and then treated with CAP for four weeks. We observed the changes of body weight and wet weight of the rat visceral fat. The plasma glucose (GLU), triglycerides (TG) and total cholesterol (TC) were assayed with enzymic method. Low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were assayed with selective precipitation method. Results: The body weight of CAP treated groups was lower than that of the HD group. The wet weight of visceral fat of HD+10 group was lower than that of the HD group, and the wet weight of visceral fat and the visceral fat/body weight of HD+20 were lower than that of the HD group (P<0.01). Compared with the HD group, the plasma TC, LDL-C, TG and GLU were reduced in CAP treated groups, while the HDL-C in these three groups were increased (P<0.01), and there was a dose dependent relationship in the CAP treated groups. However, there was no significant difference of each index between HD+20 group and ND group. Conclusion: CAP may reduce the body weight, inhibit the concentration of body fat, and regulate the level of plasma lipids and glucose in overweight rats. Therefore, CAP has a beneficial effect on the prophylaxis of the alimentary obesity.
Keywords: nutriology; capsaicin; prophylaxis; overweight; alimentary obesity; rat

Title: Research progress of the effect of Lycium barbarum polysaccharides on anioxidation and improving diabetic blood glucose and lipid metabolism

Authors: LIU Fukang, CAI Huizhen, SUN Guiju
PP: 985-991
Abstract: This paper summarized the function of Lycium barbarum polysaccharides (LBP) on antioxidation and regulating diabetic blood glucose and lipid metabolism. LBP could protect the islet cell by increasing the body antioxidant capacity, and therefore decreased the level of diabetic blood glucose. In addition, LBP was able to regulate the body related cytokines to improve insulin resistance and lipid metabolism. This review provides theoretical guidance for clinical application in the treatment of patients with diabetes.
Keywords: nutriology; Lycium barbarum polysaccharide; review; decreasing glucose; antioxidation; improving lipid mechanism

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