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June 15,2012
Volume 5,Issue 11
Pages -
Subject Area:Animal Histology and Embryology,Basic Subject of Animal Husbandry, Veterinary Medicine,Clinical Diagnostics,Health Education,Biopharmaceutics

Title: The distribution and developmental changing of IL-21 in silkie immune organs

Authors: LIU Zhiwei, PENG Kemei, SONG Hui, LIU Huazhen, CHEN Min, LU Zhiqiang, LIU Yinxue
PP: 1094-1100
Abstract: Objective: To investigate interleukin- 21 (IL- 21) distribution characteristics and development changes in silkie immune organs, in order to provide the theoretical basis for breeding and disease control and prevention. Methods: The paraffin section, immunohistochemical techniques were used in the present study. Results: 1) In the thymus of silkie, IL- 21 is mainly distributed in medulla with a small amount in cortex. Around the thymus corpuscle in medulla, there are many positive cells which are brown, round, similar in size and with no obvious nucleus. 2) In silkies' spleen, IL- 21 is mainly distributed in white pulp with a small number in red pulp. More positive cells can be seen around the spleen summary in the white pulp. Positive cells can also be observed near the marginal zone and ellipsoids of the white pulp. The round or nearly round positive cells are brownish- yellow with a smaller diameter than those in the thymus and no obvious nucleus. 3) In silkies' bursa, IL- 21 is mainly distributed in mucosal epithelium, epithelial plexus and medulla with a small amount in cortex and some in lamina propria. The brownish- yellow positive cells differ in diameter and have no clear nucleus. Conclusion: IL- 21 is a kind of cell factor that plays an important role in silkies' immune system and has a wide distribution in the immune organs. The amount of IL- 21 varies in different immune organs and in silkies of different day ages.
Keywords: livestock histology embryology; interleukin- 21; silkie; immune organ; distribution; developmental changing

Title: Prokaryotic expression, purification and polyclonal antibody preparation of TRPV6 gene in laying hen

Authors: ZHENG Yanling, HOU Jiafa
PP: 1087-1093
Abstract: To prepare the polyclonal antibody of transient receptor potential vanilloid receptor 6 (TRPV6) protein in laying hen, a pair of primers was designed and synthesized to clone the 1 801 to 2 176 position of TRPV6 gene about 375 bp by RT- PCR from the total RNA, which was extracted from ovarium. The amplified segment was inserted to the expression plasmid pET- 32a(+), then the prokaryotic expression vector pET- 32a(+)- TRPV6 was constructed and transformed into BL21(DE3) and induced by IPTG. The recombination proteins were purified by using Ni2+- NTA chelating column. The purified proteins were inoculated into New Zealand adult rabbits to develop polyclonal antibody of rabbit against chicken TRPV6. ELISA and Western blotting were then performed to evaluate the specificity of the prepared antibody. The results showed that the prokaryotic expression vector pET- 32a(+)- TRPV6 was successfully constructed in this experiment and the clone recombinant protein relative molecular weight was 35 ku detected with polyacrylamide gel electrophoresis. Western blotting indicated that the TRPV6 fusion protein displayed great immunobiological activity and anti- TRPV6 polyclonal antibody, which was purified by immunizing rabbit could bind with TRPV6 protein specially expressed in E. coli and natural TRPV6 protein extracted from long bone. The titer of antiserum generated was 1∶100 000 by ELISA. Therefore, the TRPV6 fused protein and polyclonal antibody obtained could play an important role in studying the function and mechanism of TRPV6 calcium ion channel in the formation of medullary bone in laying hen.
Keywords: clinical veterinary medicine; polyclonal antibody; prokaryotic expression; purification; TRPV6; laying hen

Title: Expression, purification and antimicrobial activity analysis of antimicrobial peptide originated from intestine of grass carp

Authors: SHI Yuting, SUN Kejun, TAO Xugu, SU Jianming, CAO Rong, CHEN Tao
PP: 1081-1086
Abstract: According to the amino acid sequence of intestine of grass carp, antibacterial peptide (AP) coding genes were synthesized artificially and cloned into vector pET32a to construct a recombinant plasmid termed pET- AP which contains a 6譎is tag. Plasmid pET- AP was transferred to E.coli strain BL21. SDS- PAGE analysis of recombinant protein was carried out after induced by IPTG. Sequencing results indicated that the sequence and location of insert fragment in pET- AP were correct. Recombinant protein was purified by Ni2+- NTA column affinity chromatography. And enterokinase was used to recover the recombinant AP. Antimicrobial activity assay showed that recombinant AP could inhibit the growth of E.coli.
Keywords: veterinary medicine; antibacterial peptide; recombinant expression; purification; antimicrobial activity

Title: Expression and function identification of PRRSV sialoadhesin fragments

Authors: XIAO Yihong, QIU Hongkai, LIU Yue, CUI Ran, ZHOU Enmin
PP: 1075-1080
Abstract: To identify the relationship of sialoadhesin and envelope glycoprotein GP5, 5 fragments of porcine sialoadhesin (pSn) were cloned from porcine alveolar macrophage (PAM) by RT- PCR. After all the fragments were cloned into pET- 28a vector and expressed. The purified proteins were identified by SDS- PAGE and Western blotting. The relationship of fragments proteins and fragments of GST- GP5 were analyzed by ELISA. The results showed that all the fragments were successfully cloned and expressed identified by SDS- PAGE and Western blotting. The binding of pSn with GP5 was confirmed by identification of ELISA. The results of ELISA showed that fragment of pSn- 3 could bind with GP5- 1 which could help to learn more about the infection mechanism of porcine reproductive and respiratory syndrome virus (PRRSV).
Keywords: veterinary medicine; porcine reproductive and respiratory syndrome virus; sialoadhesin; GP5; expression; function

Title: Analysis on regulation of gga-miR-92 on target gene ATRX

Authors: WANG Xiao, MIAO Nan, GONG Ping, PENG Xiuli, GONG Yanchang, FENG Yanping
PP: 1070-1074
Abstract: To study the regulation effect of gga- miR- 92 on chicken gonadal development, targets related to gonadal development were first predicted by using bioinformatics methods, then the regulatory relation of miR- 92 with target was identified by dual luciferase reporter assay in chicken DF- 1 cell. The results showed that alpha thalassemia/mental retardation syndrome X- linked (ATRX) was predicted to be a target of gga- miR- 92, and recombined expression vectors of ATRX 3′- UTR were constructed successfully. Dual luciferase reporter assay showed that miR- 92 decreased the activity of luciferase co- transfected with ATRX vector. It can be concluded that ATRX might be negatively regulated by gga- miR- 92.
Keywords: poultry science; chicken; miR- 92; alpha thalassemia/mental retardation syndrome X- linked; gonadal development

Title: Role of podocalyxin and TGF-β1 in patients with diabetic nephropathy

Authors: XING Lingling, YU Lianying, FU Shuxia, YANG Lin, LI Shaomei, LIU Qingjuan
PP: 1065-1069
Abstract: Objective: To investigate the relation between podocyte marker protein podocalyxin (PCX) and transforming growth factor- β1 (TGF- β1) in patients with diabetic nephropathy (DN). Methods: Twenty- four hour urinary samples from 30 patients with DN and 10 healthy volunteers as control group were collected to detect the level of PCX by ELASA. All patients with DN were divided into three groups by glomerular lesion and 10 cases of renal tumors distal renal tissue as control group. The expression of TGF- β1 in glomeruli was detected by immunohistochemistry. Results: The levels of PCX were significantly higher in the urine from patients with DN compared with those in the control group ［(32.3±3.5) mg/L vs (2.0±0.4) mg/L, P＜0.05), and gradually increased along with glomerular lesion aggravating. The expression of TGF- β1 in glomeruli was obvious in the third grade glomerular lesion, especially in sclerotic area. There were obviously positive correlation between the level of urine PCX and expression of TGF- β1 in diabetic glomeruli, 24h urine protein excretion, serum creatinine (r=0.835 P＜0.01, r=0.742 P＜0.01, r=0.599 P＜0.01, respectively). Conclusion: The level of urine PCX can be used as a novel marker for forecasting the condition of patients with DN and TGF- β1 correlates with the podocyte injury.
Keywords: medicine; podocalyxin; transforming growth factor- β1; diabetic nephropathy

Title: Glucagon-like peptide-1 prevents Alzheimer's disease-like changes in hippocampus of a rat model with type 2 diabetes

Authors: YANG Yan, JIANG Teng, HU Shuhong, YUAN Gang
PP: 1058-1064
Abstract: Objective: This study intervened glucagon- like peptide- 1(GLP- 1) on type 2 diabetes (T2D) rats, detected the insulin signaling pathway in the brain before and after treatment, and tested whether GLP- 1 could relate to decrease the risk of Alzheimer's disease (AD) in T2D. Methods: The models of T2D rats were fed with high glucose, high fat and high protein for 8 weeks, and then injected by STZ (T2D). Liraglutide- GLP- 1 analogues were subcutaneous injection for 4 weeks in T2D rats (T2D+GLP- 1). The level of plasma insulin, plasma glucose, total tau protein, the phosphorylation of tau at individual phosphorylation sites, the activities of phosphatidylinositol 3 kinase/protein kinase B (PI3K/AKT), and glycogen synthase kinase- 3β (GSK- 3β) were detected. Results: Plasma glucose, insulin and insulin resistance levels in T2D group were obviously higher than those in CTL group. Moreover, those indexes were gradually decreased after GLP- 1 subcutaneous injection, and the decrease became significant in week 4 between T2D and T2D+GLP- 1. The level of phosphorylated tau protein at site Ser199/202 and Thr217 in hippocampus of T2D rats were found to be raised notably compared with CTL group, and evidently decreased after GLP- 1 intervention. There were decreased activities of PI3K/AKT and increased activities of GSK- 3β in hippocampus of T2D rats which indicated an impaired insulin signaling pathway, while there were a rise of PI3K/AKT activity and a decline of GSK- 3β activity after 4 weeks intervention of GLP- 1. Conclusion: Tau protein was hyperphosphorylated in hippocampus of T2D rats, and these AD- like changes could be reversed after GLP- 1 intervention for 4 weeks. GLP- 1 might reduce AD- like changes in brain of T2D rats by two ways: 1) GLP- 1 could activate brain insulin signaling pathway in T2D, therefore, it might improve the AD- like changes by down- regulating the activities of GSK- 3β, which acts as a phosphorylated kinase of tau protein. 2) GLP- 1 could improve AD- like changes in T2D brain by reducing peripheral insulin resistance.
Keywords: internal medicine; glucagon- like peptide- 1; type 2 diabetes; Alzheimer's disease; insulin signaling pathway; glycogen synthase kinase- 3β

Title: Expressions of progesterone receptor and correlations with tumor necrosis factor-α in breast cancer

Authors: YU Mingxia
PP: 1054-1057
Abstract: Objective: To determine the expression of progesterone receptor (PR) in breast cancer and its correlation with tumor necrosis factor- α (TNF- α). Methods: Strept avidin- biotin complex (SABC) method was used to detect the expressions of PR and TNF- α in 112 cases of primary breast cancer, and analyzed the association of PR with clinicopathological features of breast cancer and TNF- α. Results: 1) The positive expression rate of PR in breast cancer was 23.21%（26/112）. 2) There were no significant associations of PR with TNM staging, age, age of menarche, menopause, gravidity, parity and family history of cancer. 3) PR had a positive correlation with TNF- α in breast cancer (P<0.05). Conclusion: There was no significant association of PR with clinicopathological features of breast cancer. PR had a positive correlation with TNF- α in breast cancer, but the specific mechanism needs follow- up research.
Keywords: laboratory diagnostics; progesterone receptor; tumor necrosis factor- α; breast cancer

Title: Effect in vivo of TCRP1 mediating resistance to cisplatin in OSCC

Authors: GU Yixue, HE Zhimin
PP: 1047-1053
Abstract: TCRP1 is a novel gene which was cloned from a multi- drug resistant cell line Tca8113/PYM in previous research. The results of bioinformatics indicated that TCRP1 was located on human chromosome 11q13.4, and encoded a putative protein of 235 amino acids with molecular weight of 25 ku. The preliminary functional investigation indicated that TCRP1 was a new chemo- resistance related gene, which mediated specifically resistance to cisplatin in oral squamous cell carcinoma (OSCC) cells. This paper identified the effect in vivo and the possible mechanism through establishing the ectopic tumor animal model of over/silence expression of TCRP1.
Keywords: oncology phymatology; oral squamous cell carcinoma; cisplatin; resistance; TCRP1; apoptosis

Title: Preliminary study on labeling natural regulatory T cells with superparamagnetic iron oxide in vitro

Authors: CHEN Weiwei, YU Daowu, XIONG Wei, MU Ketao, QI Jianpin, ZHU Wenzhen
PP: 1039-1046
Abstract: Objective: To label the regulatory T cells with superparamagnetic iron oxide (SPIO) and evaluate the biological effects of SPIO on the regulatory T cells in vitro. Methods: The natural regulatory T cells were isolated from the spleen of mice with EasySep Mouse CD4+ T cell pre- enrichment kit and Mouse CD25 Selection kit. The cells were then incubated with SPIO for 2 h at the iron concentration of 0.01, 0.05, 0.10 and 0.15 g/L. Cells incubated without SPIO served as the control. The uptake capability of SPIO was evaluated by Prussian blue staining and measured quantitatively with atomic absorption spectrometry. The cell viability was monitored by MTT method. The suppressive effect of regulatory T cells on the proliferation of CD4+ T cells was analyzed in proliferation assays. Results: SPIO was uptaken by regulatory T cells in a concentration- dependence manner. Within a certain range of the concentration, SPIO had no obvious effects on the viability, phenotype and suppressive function of the regulatory T cells. The MR images showed the significant decrease of magnetic resonance (MR) signal in T2 weighted images resulted from SPIO labeling of cells. Conclusion: the regulatory T cells were successfully labeled with SPIO in vitro. In a certain range of concentration, SPIO have no effects on phenotype, viability and function of regulatory T cells.
Keywords: medical imaging; superparamagnetic iron oxide; magnetic resonance imaging; regulatory T cells; in vitro

Title: Effect of human cytomegalovirus infection on secretion of RANTES in the cultured human fibroblasts

Authors: ZHANG Yan
PP: 1035-1038
Abstract: Objective: To investigate the effect of human cytomegalovirus (HCMV) infection on the expression of regulated on activation, normal T expressed and secreted (RANTES) in human embryonic lung fibroblast (HELF) so as to explore the possible mechanism of the immune- pathological changes caused by HCMV infection. Methods: Expression of RANTES mRNA was detected by RT- PCR. Secretion of RANTES protein was quantitated by ELISA. Results: After HCMV infection, the expression of RANTES mRNA was occurred at 8 h.p.i, with a maximum at 24 h. p.i, since then it remained relatively high level. The expression of RANTES protein peaked at 24 h. p.i, then dropped sharply and was almost undetectable by the time at 72 h.p.i. Conclusion: HCMV may down- regulate extra- cellular production of RANTES protein via mimicking the receptor of β family chemokine as a mechanism to evade the local immune response to infection.
Keywords: prenatal medicine; cytomegalovirus; chemotactic factors; immune response

Title: Research on relationship between social intelligence and well- being of college students

Authors: LIANG Xiaoling, MIAO Yuanjiang, MAO Longfei
PP: 1031-1034
Abstract: Objective: To explore the relationship between social intelligence and well- being. Methods: The paper investigated 205 undergraduates of Nanchang University with the Tromso social intelligence scale (TSIS) and multiple happiness questionnaire (MHQ). Results: Social intelligence was significantly associated with well- being. Social intelligence directly affected the well- being of students. Conclusion: Social intelligence can positively predict well- being, and it is the positive impact factor for well- being.
Keywords: health education; social intelligence; well- being; college students; multiple happiness questionnaire

Title: Function of c-Myc and nuclear respiratory factor-1 in cell apoptosis

Authors: SANG Hao, ZHANG Lijuan, LI Jue
PP: 1024-1030
Abstract: Apoptosis affects the morbidity and prognosis of many diseases. Both c- Myc and nuclear respiratory factor- 1(NRF- 1) are involved in the cellular apoptotic signaling pathway. Based on the introduction of c- Myc- induced cellular apoptosis, NRF- 1- mediated mitochondria injury and the synergistic effect of c- Myc and NRF- 1 in cellular apoptosis, this paper summarizes the function of c- Myc and NRF- 1 in cellular apoptosis. Studies have indicated that c- Myc and NRF- 1 may combine cytochrome C competitively in the process of cell survival and apoptosis. The study c- Myc and NRF- 1, as well as their possible interaction may conduce to better understanding of the mechanism of cell survival and apoptosis.
Keywords: toxicology; apoptosis; reviews; c- Myc oncoprotein; nuclear respiratory factor- 1

Title: Design, synthesis and bioactivity of novel PTP1B inhibitors

Authors: TONG Yuanfeng, ZHANG Pei, YE Fei, WU Song
PP: 1018-1023
Abstract: Objective: To find novel drug candidate for diabetes, a series of compounds against protein tyrosine phosphatase 1B (PTP1B) were found by the modification of the lead compound. Methods: Starting from N- (2- tetradecanoxyphenylaminocarbonyl) benzoic acid as lead compound, a series of derivatives were designed and synthesized. The inhibitory activities of these compounds in vitro against PTP1B were evaluated. Results: 13 compounds were synthesized and the activities were evaluated against PTP1B. Conclusion: The results indicated that some of the derivatives showed similar activities against PTP1B with the lead compound and IC50 of a compound arrived at 100 nm in vitro.
Keywords: medicinal chemistry; N- (2- tetradecanoxyphenylaminocarbonyl) benzoic acid; lead compound; synthesis; protein tyrosine phosphates 1B

Title: Research on the principle and application of a novel photo- induced cross- linking of unmodied proteins

Authors: HUANG Lianqi, CHENG Biao, HUANG Kun
PP: 1011-1017
Abstract: In this paper, three methods containing traditional chemical cross- linking, affinity labeling, and newly developed photo- induced cross- linking of unmodified proteins (PICUP) are picked out to be compared. And based on the experiments results on porcine insulin (PI) and human islet amyloid polypeptide (hIAPP) in our laboratory, points of the paper are focused on the principle, application, advantages and disadvantages of the PICUP methodology. According to the results, the new method has some characteristics, such as simple- procedures, wide- range as well as short time and so on. The research progress of the method in bioscience is also discussed in this paper.
Keywords: biopharmaceutics; photo- induced cross- linking of unmodified proteins; reviews; cross- linking

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