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June 15,2014
Volume 7,Issue 11
Pages -
Subject Area:Basic Subject of Animal Husbandry, Veterinary Medicine,Animal Husbandry,Health Administration,Bone Surgery,Cardiovascular Diseases

Title: Study on the RT-LAMP as a diagnostic tool of Toxoplasma gondii in pork and its products

Authors: QU Daofeng, HAN Jianzhong, DU Aifang, LIU Xiaoning
PP: 1112-1118
Abstract: In this study, a fast, sensitive and specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the detection of Toxoplasma gondii in pork and its products was developed. We used a conserved sequence of 18S rRNA of T. gondii in GenBank to design primers for RT-LAMP test. The amplication was able to finish in 60 min under Bst DNA polymerase at 65℃ by employing a set of six primers. With the addition of the SYBR Green I, we analysized gel electrophoresis results. The assay showed 10 times higher sensitivity than RT-PCR using T. gondii RNA as template. The RT-LAMP assay was also assessed for specificity and was found to precisely discriminate between positive and negative test samples. As RT-LAMP requires very basic instruments and the results can be obtained by visual observation, this technique provides a simple and reliable tool for inspecting food which are T. gondii-contaminated.
Keywords: other subjects of animal husbandry and veterinary medicine science; Toxoplasma gondii; reverse transcription loop-mediated isothermal amplification; 18S rRNA; detection; pork and its products

Title: Seasonal changes of SOD activity in the epididymis of the soft-shelled turtle (Pelodiscus sinensis)

Authors: BIAN Xunguang, YANG Ping, CHEN Qiusheng
PP: 1108-1111
Abstract: In order to study the variation of superoxide dismutase (SOD) in epididymis of soft-shelled turtle during the reproductive cycles, the SOD activity in both tissues and fluid in each segment of epididymis were measured during reproductive seasons (June and August) and hibernation seasons (December and February). The results showed that the SOD activity in epididymal fluid had varied among each segment of epididymis throughout the reproductive cycles, and was higher in reproductive seasons than that in hibernant seasons at same segment. It showed that the highest SOD activity in caudal epididymal fluid was (1 518.53 24.88) U/mL, extremely striking contrast with it in caput and corpus (P<0.05) during reproductive seasons. However, in hibernant seasons, the SOD activity in caput epididymal fluid was the highest with (508.45 45.40) U/mL, and it was significant different with that in corpus and cauda (P<0.05). Among epididymal tissue, the SOD activity had little discrepancy, and had the highest level at cauda with (137.58 4.81) U/mg prot, which was significantly different with that in corpus (P<0.05), while no difference with that at caput (P>0.05) during reproductive seasons. The SOD activity in epididymal tissue had no significant contrast among each segments (P>0.05) in hibernation seasons, and was lower than that in reproductive seasons. In the epididymis, the SOD activity showed seasonal variation, suggesting that it might have a certain correlation with the movement and storage of sperm in the epididymis.
Keywords: basic subject of animal husbandry and veterinary medicine science; soft-shelled turtle; epididymis; superoxide dismutase

Title: Expression patterns of let-7c and miR-23a in large white pig testes and miRNA expression vector construction

Authors: ZHOU Jiawei, YE Lianzhi, LUO Lifan, GUAN Kaifeng, MA Changping, XIA Xuanyan, LEI Bin, LI Feng’e
PP: 1102-1107
Abstract: This study amplified and cloned let-7c and miR-23a, and obtained their precursor and mature sequences of miRNA. RT-PCR was used to analyze the expression pattern of let-7c and miR-23a in the porcine testes at age of 35, 60, 90, 180 d. The results showed that pri-let-7c had the highest expression at 180 d, but scarcely expressed at 60 d, while mature let-7c had a lower expression at 180 d than other age groups (P<0.05). pri-miR-23a highly expressed at 35 d and 90 d, while the mature miR-23a had a high expression at 90 d than at 35 d and 180 d (P<0.05). Using online RNAhybrid and RNA22, sperm adhesion molecule 1 (SPAM-1) and TNF receptor superfamily member 6 （FAS） genes were predicted to be the target genes of let7c and miR-23a, respectively. plet-7c and pmiR-23a expression vectors were constructed by inserting pri-miRNA fragment in, and then transfected pig swine testicular (ST) cells. RT-PCR was used for expression confirmation and obtained the corresponding expression information.
Keywords: animal husbandry; miRNAs; RT-PCR; pig; spermatogenesis

Title: Isolation and identification of three diterpenoid compounds extracted from Jiyuan rubescens

Authors: WANG Shubo, FU Ting, WANG Fanhua, HOU Ruijuan, LI Huiju, KE Yu, LIU Hongmin
PP: 1097-1101
Abstract: Objective: To study the chemical constituents of Jiyuan rubescens. Methods: The chemical constituents were isolated by method of chromatographic isolation, and their structures were elucidated by the analysis of spectral data and physiochemical properties. Furthermore, their cell cytotoxicity was also tested. Results: Three diterpenoid compounds were isolated from Jiyuan rubescens. Conclusion: These three compounds were firstly isolated from Jiyuan rubescens, in which compounds 1 and 3 showed certain cytotoxicity on human esophageal carcinoma Eca109 cells.
Keywords: medicinal chemistry; Jiyuan rubescens; chemical composition; structure identification; diterpenoid

Title: Model establishment and application of PAMPA for blood-brain barrier

Authors: MA Linnan, LIU Hui, NI Jingman
PP: 1092-1096
Abstract: Objective: The aim of the study was to identify a model for the blood-brain barrier based on the use of the parallel artificial membrane permeation assay (PAMPA) model, and to predict the blood-brain barrier permeability of a set of test compounds. Methods: The filter membrane was coated with porcine polar brain lipid (PBL) in dodecane and effective permeability of the compounds was calculated by using high performance liquid chromatography (HPLC). Results: The filter membrane was coated with 5 μL PBL in dodecane, the concentration of DMSO was below 5%, and incubated for 16 h at 25℃ while the permeation occurred. On this condition, a steady and reliable artificial membrane assay was developed. Conclusion: The PAMPA has the advantages of predicting passive blood-brain barrier penetration with low cost and reproducibility.
Keywords: pharmacy; parallel artificial membrane permeation assay; blood-brain barrier; model establishment

Title: Rules of the USA medical device unique identification database and inspiration

Authors: LIU Qingfeng
PP: 1087-1091
Abstract: In order to offer inspiration for medical devices enterprises and Chinese regulatory government for medical devices, this paper discussed the concept and the implementation significance of unique device identification (UDI), and made an induction of the UDI final rule issued by Food and Drug Administration (FDA) of the USA and introduced the relevant rules issued by the FDA about medical device unique device identification database (UDID). The results showed that China should accelerate the legislation pace of regulatory for medical devices, and related enterprises should learn and apply UDI rules as soon as possible.
Keywords: health management; unique device identification database; rule; inspiration

Title: Effect of irradiation on the multi-drug resistance of ovarian cancer cell SKVCR2.0

Authors: XU Shan, KONG Dejuan, WANG Hongzhi, CHEN Haiyang, MA Shumei, LIU Xiaodong
PP: 1080-1086
Abstract: Objective: To discuss the effect of irradiation on the multi-drug resistance (MDR) of ovarian cancer cells. Methods: MTT assay was used to detect the drug sensitivities of SKVCR2.0 and SKOV-3 cells in vincristine (VCR), cis-dichlorodiamineplatinum(II) (DDP) and therarubicin (THP) groups. We chose ovarian cancer cells SKVCR2.0 induced by VCR and then irradiate them. The methods of irradiation included conventional fraction (2 Gy/time, 1 time/d) and hyperfraction (1 Gy/time, 2 times/d, with 4 h interval). The total dose was 6 Gy. The viability of SKVCR2.0 cells was tested by cell counting kit-8 (CCK-8), and the incidence of autophagy of SKVCR2.0 cells was tested by monodansylcadaverine (MDC). Then, based on the above, the cell viability and the incidence of autophagy after adding three methyl adenine (3-MA) were tested. Results: Compared with SKOV-3, SKVCR2.0 had higher MDR. With the increase of VCR, the cell viability of SKVCR2.0 decreased, while after irradiation, cell survival increased and the incidence of autophagy increased significantly. Moreover, there was a higher incidence of autophagy in hyperfraction group. In addition, when adding 3-MA, the incidence of autophagy of the two groups decreased. Conclusion: Fractionated irradiation leads to MDR through increased the incidence of autophagy.
Keywords: radiohygiene; ovarian cancer cell; irradiation; multi-drug resistance

Title: Study on effect of silicate bacteria metabolites on the experimental asbestosis

Authors: WANG Ye, YANG Chenlu, CHENG Weibo, LIU Hui, ZHAO Yujia, LIU Huiran
PP: 1071-1079
Abstract: Objective: To detect the protective effect of the silicate bacteria metabolites on the rats that have inhaled the asbestos. Methods: 80 healthy male rats were divided into normal control group (n=5), treatment control group (n=5), asbestos-inhaled group (n=35) and asbestos-inhaled-and-treated group (n=35). The asbestos-inhaled group and asbestos-inhaled-and-treated group were also divided into 7 subunit groups i.e 1 d, 3 d, 1-week, 2-week, 4-week, 8-week, 10-week group, 5 rats in each group. For each subunit group of asbestos-inhaled group, the asbestos lung model was established by using the method of ultrasonic atomization asbestos dust. For treatment group, besides for model establishment, at the same time, the rats were treated drug with ultrasonic atomization metabolites of silicate bacterium. For normal group, the rats were only with the inhalation of nebulized distilled water. For treatment and control group, the rats were only with the inhalation of the atomization of silicate bacteria metabolites. Each group rats were sacrificed at the corresponding day. The lung and blood samples were extracted for pathological examination. The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and hydroxyproline (HYP) were detected. The results were analyzed by statistics software. Results: Normal control group looked normal. Treatment control group was similar to the normal control group, and slight inflammatory reaction could be seen occasionally. Mild acute inflammation could be seen in the asbestos-inhaled group at early stage, then it converted to chronic inflammation with fibrous tissue hyperplasia. As time went on, the fibrosis became more serious. In the asbestos-inhaled-and-treated group, inflammatory reaction was very slight and only a little fibrous tissue hyperplasia could be seen. There were no significant difference of the levels of the MDA, SOD, HYP between the normal control group and treatment control group, as well as the lung index. However, the SOD, MDA, HYP of asbestos-inhaled group and asbestos-inhaled-and-treateel group were higher than those of normal control group, while the MDA, HYP of asbestos-inhaled-and-treated group were lower than those of asbestos-inhaled group, but SOD was higher in the former. Conclusion: The silicate bacteria metabolites could slow down the development of asbestosis in a certain extent.
Keywords: occupational medicine; treatment of asbestosis; silicate bacterial metabolites; experimental research

Title: Effect of retinoic acid on the expression of CD2AP gene mRNA level

Authors: QIU Lingzhi, GAO Shan, ZHU Lianghua, ZHUANG Lili, ZHOU Guoping
PP: 1066-1070
Abstract: Objective: To investigate the effect of 9-cis retinoic acid (9-cis RA), 13-cis RA and tamibarotene (AM80) on the expression of CD2AP (CD2-associated protein) gene mRNA level. Methods: Total RNA prepared from HK-2 and HEK-293 cells, with or without 9-cis RA,13-cis RA, AM80, was subjected to real-time PCR (RT-PCR). Results: The level of CD2AP transcripts increased with 9-cis RA treatment in HK-2 and HEK-293 cells. But 13-cis RA and AM80 could not increase the expression of CD2AP gene mRNA level in both HK-2 and HEK-293 cells. Conclusion: 9-cis RA may have a therapeutic effect on the kidney disease with a decreased expression of CD2AP, while the 13-cis RA, AM80 have no similar effect.
Keywords: pediatrics; retinoic acid; CD2AP; mRNA

Title: In situ cartilage repair using NCP/PLLA scaffold incorporated with bFGF

Authors: HUANG Xin, SHI Zhongli, WENG Wenjian, YANG Disheng
PP: 1061-1065
Abstract: Objective: To evaluate the effort of the novel nano-α/β calcium phosphate (NCP) /poly-L-lactic acid (PLLA) incorporated with basic fibroblast growth factor (bFGF) as the carrier for articular cartilage regeneration in vivo. Methods: Full-thickness defects (diameter 4 mm×4 mm in depth) were created in the medial femoral condyles in 12 adult male rabbits. The defect in one side of condyle was filled with NCP/PLLA scaffold with bFGF mixture and another was left without treatment as a control. Macrography and histological analysis were evaluated after 4 and 8 week postoperation. Results: At 4 week postoperation, the defects in experimental group were filled with white regenerated tissue. Histological analysis showed that abundant of undifferentiated cells or immature cartilaginous cells were present in the micropores with clusters of cartilaginous extracellular matrix formation. At 8 week postoperation, well-formed and mature cartilage was resurface the defects. Whereas only fibrous tissue replacement was observed for the control either at 4 or 8 weeks. Conclusion: The novel NCP/PLLA incorporated with bFGF can effectively treat the cartilage defects.
Keywords: bone surgery; cartilage regeneration; tissue engineering; basic fibroblast growth factor

Title: Study on the synergic action of human herpes simplex virus and inorganic mercury in animal model

Authors: XIONG Tianqing, TAN Baihong, LI Yanchao
PP: 1055-1060
Abstract: In order to investigate the possible synergic mechanism of inorganic mercury and human herpes simplex virus in the central nervous system, about 75 μL HSV-1 (2 × 106 PFU) and 75 μL 2 g/L HgCl2 was injected bilaterally into the footpads of 14-week old C57BL/6 for 2 d, then 75 μL HSV-1 (2×106 PFU) was injected into the footpads. The positive neurons for mercury were detected by autometallography. Immunofluorescence was used to reveal the changes of Tau protein and phosphorylation of Tau protein. The results showed that after footpad injection, mercury was found within the motoneurons in the spinal anterior horn after the 4th day of injection. As compared with the control group and mercury-injected group, Tau protein and phosphorylated Tau protein were significantly increased in the spinal cord in the group which was injected with both mercury and HSV-1. These findings suggested that there might be a synergic mechanism between mercury and HSV-1, which was related to the neurodegenerative changes in the central nervous system.
Keywords: neurology; human herpes simplex virus; inorganic mercury; animal model; Tau protein

Title: A potential therapeutic effect of arachidonic acid catalysed by cytochrome P450 epoxygenases on anti-LPS activity

Authors: LIU Wanjun, ZENG Hesong
PP: 1049-1054
Abstract: Objective: To test mechanisms for that arachidonic acid catalysed by cytochrome P450 (CYP) epoxygenases resistance to the damage and expression of inflammatory cytokines induced by lipopolysaccharides (LPS) in human umbilical vein endothelial cells (HUVECs). Methods: In our study, we used CYP2C8 gene transfection and direct epoxyeicosatrienoic acids (EETs) intervention before LPS infused in HUVECs, then examined the expression of NF-κB and the apoptsis related protein Bcl-2, Bax by Western blotting and apoptosis by flow cytometry examination. Results: CMV-CYP2C8 transfection and direct intervention of EETs markedly suppressed LPS induced inflammatory cytokines IL-6 and MCP-1 expression via activation of NF-κB. In addition, the expression of anti-apoptosis proteins Bcl-2 decreased and the pro-apoptosis proteins Bax increased. Moreover, after using of peroxisome proliferator-activated receptor γ (PPARγ) inhibitors, these effects were partially inhibited. Conclusion: Our results demonstrated that CYP2C8-derived EETs had anti-vascular inflammatory and anti-apoptosis effects, at least in part, through activation of PPARγ.
Keywords: cardiology; arachidonic acid cytochrome P450 epoxygenase; atherosclerosis; lipopolysaccharide; inflammation; apoptosis

Title: Multidrug resistance-reversing effect of epigallocatechin gallate on human nasopharyngeal carcinoma cell lines by decreasing expression of P-glycoprotein

Authors: ZHU Meichan, XIE Mao, SHI Shujing, LI Xiaoxue, QI Chenglin, LI Heng, YI Xiang, TANG Anzhou
PP: 1044-1048
Abstract: Objective: To evaluate the value of epigallocatechin gallate (EGCG) on the multi-drug resistance (MDR) reversing effect human nasopharyngeal carcinoma (NPC) HNE1/DDP cell lines and its possible mechanisms. Methods: Cytotoxicity of EGCG and its sensibility to cis-platinum in HNE1/DDP cell line, were evaluated by MTT assay. Furthermore, the expression of P-glycoprotein (P-gp) was detected by quantitative real-time PCR (QRT-PCR) and Western blotting respectively. Results: The 10% inhibition concentration of EGCG to HNE1/DDP cell line was 154.6 μmol/L. Therefore, 60, 80, 120 μmol/L of EGCG were used to detect the reversal effect of MDR. 50% inhibition concentration (IC50) for DDP was 4.22 mg/L. EGCG could increase the drug susceptibility of HNE1/DDP cell to DDP, and the IC50 for DDP decreased to 3.82, 2.56, 1.78 mg/L, and the modulating factors were 1.10, 1.65 and 2.37 fold respectively. The expressions of P-gp in HNE1/DDP cells incubated with EGCG were deceased when detected by QRT-PCR and Western blotting. Conclusion: EGCG could partially reverse the multidrug resistance of the NPC HNE1/DDP cells in vitro. Moreover, the reversal effect may be due to the increase of intracellular accumulation of chemotherapeutic drugs and the decrease of the expression of P-gp.
Keywords: oncology; nasopharyngeal carcinoma; epigallocatechin gallate; P-glycoprotein

Title: Effects and mechanisms of suppressing Hedgehog signaling pathway on proliferation in colorectal cancer cells

Authors: WANG Jun, DONG Weiguo, GUO Xufeng, SONG Jia, YU Shijie, LIU Min
PP: 1038-1043
Abstract: Objective: To explore the effect of Hedgehog (Hh) signaling pathway in colorectal cancer cells and its mechanism. Methods: Human colorectal cancer LoVo cells were selected and intervened by inhibitors cyclopamine and GANT61, respectively. The MTT colorimetry was used to detect the growth inhibitory rates, and the apoptosis potential of cells was assessed by flow cytometry. In addition, Hochest33258 fluorochrome staining was employed to observe cell apoptosis and the expressions of Hedgehog signaling related protein such as Shh, PTCH, Smo and Gli-1and apoptosis proteins were detected by Western blotting. Results: Cyclopamine and GANT61 inhibited the growth of LoVo cells in a dose- and time-dependent manner. Compared with the cyclopamine group, the inhibitory rate of GANT61 group was higher. The apoptosis rates also increased after drug intervention. Hochest33258 fluorochrome staining showed that there were many apoptosis cells in cyclopamine and GANT61 groups. The expression of Gli-1 and Bcl-2 were decreased, while the expression of Bax was increased. Conclusion: Hh signaling pathway was activated in human colorectal cancer cells. Inhibiting this pathway, especially specific blocking Gli-1, can inhibit cell proliferation and induce apoptosis, the mechanism may be that the nuclear transcription factor Gli-1 of Hh signaling pathway downstream is blocked, thus causes Bcl-2 down-regulation and Bax up-regulation.
Keywords: oncology; colorectal cancer; Hedgehog signaling pathway; Gli-1; proliferation; apoptosis

Title: TRB3 mediates apoptosis of renal tubular cells in diabetic nephropathy

Authors: WANG Weiwei, LÜ Shasha, CHENG Jing, LIU Xiangchun, GUAN Guangju
PP: 1031-1037
Abstract: Objective: To research the expression of tribble 3 (TRB3) in the kidney of diabetic nephropathy (DN) rats and the relationship between TRB3 expression and apoptosis of renal tubular cells. Methods: 40 male Wistar rats were randomly divided into the control group, diabetes mellitus (DM) group, DM+vehicle group and DM+TRB3-siRNA group. The DM rat model was induced by streptozotocin (STZ) injection in abdominal cavity, and intravenous injection of lentivirus silenced the TRB3 expression. The body weight, blood glucose and urinary albumin excretion (UAE) were regularly monitored.The immunohistochemical, RT-PCR and Western blotting were used to check TRB3 expression in kidney. In situ terminal deoxynucleotidyl transferase-dUTP nick-end labeling (TUNEL) and Caspase 3 activity measurement were used to check the apoptosis of tubular cells. Results: Compared with the control, TRB3 expression was obviously upregulated in kidneys of diabetic rats. Immunohistochemical staining localized TRB3 expression in renal tubules, but not in glomeruli. TUNEL staining showed that apoptotic cells were rarely observed in kidneys of control rats, but significantly increased in tubules of diabetic kidneys. In addition, Caspase 3 activity was markedly higher in kidneys of diabetic rats compared with the control. However, the number of apoptotic tubular cells and Caspase 3 activity were significantly decreased after TRB3 silencing. Conclusion: TRB3 is upregulated in kidneys of diabetic rats and mediates apoptosis of renal tubular cells.
Keywords: internal medicine; diabetic nephropathy; apoptosis; gene silencing; TRB3

Title: Protection of luteolin on rat cardiomyocytes following ischemia/reperfusion through ERK/PP1a/PLB/SERCA2a/JNK pathways

Authors: XU Tongda, WU Xin, LI Dongye, ZHU Shasha, CHEN Qiuping
PP: 1023-1030
Abstract: Objective: In order to analysize the possible molecular mechanisms that luteolin (Lut) improves contractile function of rat cardiomyocytes during ischemia-reperfusion (I/R) through ERK1/2 and JNK pathways by the assay of cardiomyocytes shortening amplitude and some related apoptosis proteins. Methods: The hearts of the normal rats were further isolated into single ventricular myocytes to simulate the I/R process using the following groups: 1) DMSO group; 2) I/R group; 3) Lut+I/R group; 4) PD98059 pretreatment (PD+I/R) group; 5) PD +Lut+I/R group; 6) SP600125 pretreatment (SP+I/R) group. The cells were harvested for measuring of single cardiomyocyte shortening amplitude and Western blotting assay checked the expression of Bcl-2, Bax, ERK1/2, p-ERK1/2, JNK, p-JNK, PP1a, p-PP1a, PLB, p-PLB and sarcoplasmic reticulum calcium-ATPase (SERCA2a). Results: Lut and SP pretrearment could improve the cell cardiomyocyte shortening amplitude and increase the Bcl-2/Bax ration. Lut affected the expression of the shortening relevant proteins PP1a, PLB and SERCA2a. However, SP pretreatment had no effect on contract related proteins. Administration of PD before Lut significantly reversed the protection of Lut. Conclusion: The protect effect of Lut is related to the expression of Bcl-2, Bax, p-ERK1/2, p-PP1a, SERCA2a and p-PLB, at least in part, by activation of the ERK1/2 signal pathway, and thus protects the function of the heart.
Keywords: internal medicine; luteolin; ischemia/reperfusion; contraction function; ERK1/2 pathway; JNK pathway

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