2.Long non-coding RNA TTTY15 expression in osteosarcoma and its effect on viability and invasion ability of osteosarcoma cells
Qing XIE ; Ke-Qin WU ; Qi-Rong CHEN ; Ning LU
Chinese Journal of Pathophysiology 2018;34(5):819-824
AIM:To observe the expression of long noncoding RNA TTTY 15 in osteosarcoma tissues and cell lines and to explore its effect on the viability and invasion ability of osteosarcoma cell lines.METHODS:qPCR was used to detect the expression of TTTY15 in 11 cases of osteosarcoma and its adjacent tissues.The mRNA levels of TTTY15 in os-teosarcoma cell lines(143B,Saos2,MG-63,U2OS and HOS)and human osteoblast cell line hFOB1.19 were also tested. TTTY15 was down-regulated after transfected with small interfering RNA in MG-63 cells,the cell line with the highest level of TTTY15.The effect of TTTY15 knockdown on the viability of MG-63 cells was measured by CCK-8 assay.The cell cycle distribution was analyzed by flow cytometry.The effect of TTTY15 knockdown on the cell invasion ability was detected by Transwell assay.The levels of miR-216b-5p and FOXM1 mRNA were detected by qPCR, and the changes of the related proteins were determined by Western blot.RESULTS:Compared with the adjacent tissues,the expression of TTTY15 in-creased in the osteosarcoma tissues(P<0.01).Compared with the human osteoblast cell line,the expression of TTTY15 increased in the osteosarcoma cell lines(P<0.05), and the level of TTTY15 in the MG-63 cells was the highest(P<0.01).After knockdown of TTTY15 expression in the MG-63 cells,the cell viability was decreased(P<0.05),cell cycle progression was inhibited(P<0.01), and the cell invasion ability was decreased(P<0.01).The expression of miR-216b-5p was increased(P<0.01)and the expression of FOXM1 mRNA was decreased(P<0.01).The protein expres-sion of FOXM1,CDK4,cyclin D1,MMP-2 and N-cadherin was decreased,while the protein expression of E-cadherin was increased(P <0.05).CONCLUSION: The expression of TTTY15 is increased in the osteosarcoma tissues and cell lines.The low expression of TTTY15 inhibits the cell viability and invasion ability of osteosarcoma cells.The possible mechanism is that the knockdown of TTTY 15 expression results in the increase in miR-216b-5p expression and the down-regulation of FOXM1 expression.
3.Minimally invasive anatomic research of microvascular decompression for trigeminal nerve utilizing virtual reality technique
Qing ZHOU ; Ke TANG ; Jinjuan XIE ; Jing'an ZHOU
Chinese Journal of Neuromedicine 2015;14(10):1027-1030
Objective To discuss the microanatomy features of microvascular decompression for trigeminal nerve through minimally invasive approach based on virtual reality image models.Methods CT and MRI scans were performed to five adult cadaver heads, and then, image data were inputted into vitrea virtual reality system to establish three-dimensional anatomy models of cerebellopontine angle.Suboccipito-retrosigrmoidal approach was simulated by selecting asterion as osseous landmark points of craniotomy and cistern segment of trigeminal nerve as exposed target.Anatomic exposures in surgical trajectory were observed and measured before and after minimally invasive design, respectively.Results Routes simulating microvascular decompression for trigeminal nerve through suboccipito-retrosigmoidal approach passed under the inferior edge of transverse sinus.Spacial relationship among route and surrounding nerves and vessels were displayed clearly.Measurement and comparative analysis among different routes indicated the volumes of routes: route b> route c > route a, and involved cerebellar volumes: route a >route c > route b.Conclusion Minimally invasive routes of suboccipito-retrosigmoidal approach can reduce surgical injure without sacrifice of anatomic exposure in microvascular decompression for trigeminal nerve.
4.Development of peripheral neuropathy rat model induced by 1-bromopropane.
Qing-hua WANG ; Zhi-xia ZHONG ; Jing-jing CHEN ; Ke-qin XIE ; Xiu-lan ZHAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(10):751-755
OBJECTIVETo observe the peripheral neurotoxicity of 1-bromopropane (1-BP) by developing an animal model of peripheral neuropathy through oral administration of 1-BP.
METHODSForty male Wistar rats were randomly and equally divided into low-dose group (200 mg/kg), medium-dose group (400 mg/kg), high-dose group (800 mg/kg), and control group. The rats in the low-dose, medium-dose, and high-dose groups were orally given 1-BP (dissolved in corn oil), while the rats in the control group were orally given an equal volume of corn oil. The oral administration (0.2 ml/100 g BW) was performed once per day, 5 days per week, for 16 consecutive weeks. Neurobehavioral indices including gait score, hindlimb grip strength, and hindlimb landing foot splay were recorded periodically. Hematological and biochemical parameters were also measured during and after 1-BP exposure.
RESULTSThe gait scores were significantly higher in the high-dose group (after 8 ∼ 16 weeks of 1-BP exposure), medium-dose group (after 14 ∼ 16 weeks of 1-BP exposure), and low-dose group (after 15 ∼ 16 weeks of 1-BP exposure) than in the control group (P < 0.05, P < 0.01). Compared with the control group, the high-dose group showed significantly decreased hindlimb grip strength after 9, 12, and 14 weeks of 1-BP exposure (P < 0.05, P < 0.01), with the hindlimbs paralyzed after 16 weeks of 1-BP exposure. After 16 weeks of 1-BP exposure, the hindlimb grip strengths of rats in the medium-dose and low-dose groups were decreased to 72.6% and 91.2% of the control value (P < 0.01, P < 0.05). Compared with the control group, the high-dose group showed significantly increased hindlimb landing foot splay after 12, 14, and 16 weeks of 1-BP exposure, and the medium-dose group showed significantly increased hindlimb landing foot splay after 14 and 16 weeks of 1-BP exposure (P < 0.05, P < 0.01). The high-dose and medium-dose groups showed significantly higher serum alanine aminotransferase (ALT) activity than the control group after 8 weeks of 1-BP exposure, and so did the low-dose group after 16 weeks of 1-BP exposure (P < 0.01).
CONCLUSIONThe nervous system is sensitive to the toxic effect of 1-BP, and 1-BP exposure can induce peripheral neuropathy in rats.
Animals ; Disease Models, Animal ; Hydrocarbons, Brominated ; administration & dosage ; toxicity ; Male ; Peripheral Nervous System Diseases ; chemically induced ; physiopathology ; Rats ; Rats, Wistar
5.Protective effect of garlic oil given at different time against acute liver injury induced by CCl4.
Gui-li ZHANG ; Tao ZENG ; Qing-shan WANG ; Xiu-lan ZHAO ; Fu-yong SONG ; Ke-qin XIE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2010;28(3):190-194
OBJECTIVETo observe and compare the protective effect of garlic oil against carbon tetrachloride (CCL)-induced acute liver injury.
METHODSThe experiments include 4 preventive groups and 2 therapeutic groups. In every preventive and therapeutic group, the mice were randomized into 6 groups with 15 each, including one negative control group, one solvent control group, one CCl4 model group and 3 garlic oil groups (25, 50, and 100 mg/kg body weight). Before given a single gavage of CCl4 (80 mg/kg), the mice were pretreated with garlic oil by gavage in preventive group 1 (30 days, once daily), preventive group 2 (5 days, once daily), preventive group 3 (ahead of 2 h, once), preventive group 4 (immediately, once) or the vehicle (corn oil, 10 ml/kg) in solvent control group. In therapeutic groups, the mice were gavaged garlic oil 2 h (once, in therapeutic 1) or for 5 days (once daily, in therapeutic 2) after administration CCl. After 24 h of the last administration, blood was collected and centrifuged at 2500 r/min at 4 degrees C for 10 min, and serum was removed to measure ALT and AST activities. The liver was dissected, weighed to calculate the liver coefficient (relative liver weight). At the same time, the liver samples were studied by histological examinations.
RESULTSCompared with negative group, the liver coefficient and the activities of ALT and AST in serum of model group were increased remarkably (P < 0.01). Compared with CCl model group, the liver coefficient and the activities of ALT and AST in serum were decreased significantly (P < 0.01) by garlic oil dose-dependently in each preventive group. Simultaneously, histological assessment showed that garlic oil effectively alleviated hepatocyte injuries induced by CCl4. Comparing the preventive effects of garlic oil in every group, it was better in preventive group 3 than others. However, all indexes and histological examinations in therapeutic group 1 did not show the difference with those of CCl4 model group. In therapeutic group 2, all indexes recovered after 5 d of CCl4 administration.
CONCLUSIONSGarlic oil can prevent acute liver injury induced by CCl4 and the effect is better in ahead of 2 h group than others.
Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Carbon Tetrachloride Poisoning ; drug therapy ; prevention & control ; Chemical and Drug Induced Liver Injury ; drug therapy ; prevention & control ; Garlic ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Plant Oils ; administration & dosage ; therapeutic use
6.Time dependent changes in antioxidation of rat serums and sciatic nerves caused by toxicology.
Ying-jian ZHU ; Qing-shan WANG ; Li-ping ZHANG ; Xin GUO ; Ke-qin XIE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(7):394-398
OBJECTIVETo study the time dependent antioxidation changes of serum and sciatic nerve in rats intoxicated with acrylamide.
METHODSMale Wistar rats weighing 180 to 220 g were given acrylamide dissolved in physiological saline (40 mg/kg ip 3 days/week). The control groups received normal saline. The gait was observed and antioxidant indexes of rat serum and sciatic nerve were determined on 0, second, fourth, sixth, 10th week.
RESULTSWith the extension of the intoxication period, compared with the control, the contents of glutathione in serum and sciatic nerve gradually decreased (P < 0.05; after 6 and 10 weeks to 92% and 77%; after 2, 4, 6 and 10 weeks to 92%, 82%, 67% and 66%); the levels of malondialdehyde gradually increased (P < 0.05; after 4, 6 and 10 weeks to 113%, 118% and 120%; after 4, 6 and 10 weeks to 153%, 167%, 174%); the abilities of the resistance to reactive oxygen species gradually decreased (P < 0.05; after 10 weeks to 82%; after 6 and 10 weeks to 76% and 71%); the activities of glutathione peroxidase gradually increased (P < 0.05; after 2, 4, 6 and 10 weeks to 122%, 130%, 160% and 124%; after 4, 6 and 10 weeks to 134%, 152% 164%); the activities of glutathione reductase increased at early stage (P < 0.01; after 4 and 6 weeks to 300% and 217%; after 4 weeks to 142%) and decreased later (P < 0.01; 6 and 10 weeks to 59% and 33% in sciatic nerve); the activities of superoxide dismutase increased primitively (P < 0.05; after 2 weeks to 110%; after 4 weeks to 124%) and decreased later (P < 0.05; after 10 weeks to 85% in serum). The changes of antioxidant indexes in serum and sciatic nerve according to gait score were similar. The level of MDA in serum was in high correlation (P < 0.01) with that in sciatic nerve. The regression coefficients were 0.99 and 0.96 according to the administration time and gait score respectively.
CONCLUSIONThe changes of the antioxidant indexes in serum and sciatic nerve of rat treated with acrylamide are time dependent. The changes in serum and sciatic nerve are similar but those in sciatic nerve are more remarkable.
Acrylamide ; toxicity ; Animals ; Glutathione ; blood ; metabolism ; Glutathione Reductase ; blood ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Malondialdehyde ; blood ; metabolism ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; blood ; metabolism ; Sciatic Nerve ; drug effects ; metabolism ; Superoxide Dismutase ; blood ; metabolism
7.Effects of garlic oil on 2, 5-HD induced lipid peroxidation damage in rat nerve tissues.
Xu-Cong GAO ; Tao ZENG ; Qian WANG ; Qing-Shan WANG ; Ke-Qin XIE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(11):649-653
OBJECTIVETo investigate the effects of garlic oil (GO) against the peroxidation damage of rat nerve tissue and the peripheral motor neuropathy induced by 2, 5-HD.
METHODSMale Wistar rats were divided into four groups, with 10 in each group. The model group, and low and high doses of GO groups were administrated with 2, 5-HD (ip, 300 mg/kg), respectively; The control group was treated with sodium chloride, five times per week for six weeks. Pretreatment with GO gavaged (40 mg/kg or 80 mg/kg) started one week be-fore 2, 5-HD treatment, and lasted to the end of the experiment. Neurobehavioral indexes were examined at the zero, second and fourth week. At the end of the experiment, the scores of the gait, and the concentration of MDA and GSH, the level of TAOC and the ability of inhibition of.OH in cerebrum, spinal cord and sciatic nerve were examined.
RESULTSCompared with the zero week, except of the control group rats, the hind limb landing foot splay of three groups rats decreased by 44%, 50% and 49% at the fourth week, respectively without significant difference. The threshold value of balance in model, GO low and high doses groups rats decreased by 30%, 45% and 68% at the fourth week, respectively, and lower than the control group rats (P < 0.01). GO low and high doses groups rats showed the serious abnormality at the fourth week, before one week of the model group rats. The scores of gait of model, and GO low and high doses groups rats increased significantly compared with control group rats, and the GO high dose group rats were higher than model group rats (P < 0.05). Increase of the concentration of MDA, and decrease of the level of the ability of inhibition of.OH were induced by 2, 5-HD in cerebrum, spinal cord and sciatic nerve. The concentration of MDA increased, and the level of the ability of inhibition of.OH decreased (P < 0.05 or P < 0.01), respectively. The results showed that the concentration of MDA decreased, and the level of the ability of inhibition of.OH induced by GO in cerebrum, spinal cord and sciatic nerve increased, the concentration of MDA of GO low doses group rats decreased, the level of the ability of inhibition of.OH increased, the concentration of MDA of GO high doses group rats decreased (P < 0.01) respectively, and the level of the ability of inhibition of.OH increased (P < 0.01) in nerve tissue.
CONCLUSIONGO has antagonist effect on the 2, 5-HD induced peroxidation damage, but can not improve the function of the peripheral motor nerve, indicating that the lipid peroxidation does not play an important role in 2, 5-HD neurotoxicity.
Allyl Compounds ; pharmacology ; Animals ; Drug Antagonism ; Garlic ; chemistry ; Hexanones ; toxicity ; Lipid Peroxidation ; drug effects ; Male ; Nerve Tissue ; drug effects ; metabolism ; pathology ; Plant Oils ; pharmacology ; Rats ; Rats, Wistar ; Sulfides ; pharmacology
8.Dynamic study of neurofilament contents in rat's spinal cord induced by 2, 5-hexanedione.
Fu-Yong SONG ; Qing-Shan WANG ; Tao ZENG ; Li-Hua YU ; Zhen-Ping ZHU ; Ke-Qin XIE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(10):588-591
OBJECTIVETo investigate the dynamic changes of neurofilament contents in rat's spinal cord induced by 2, 5-hexanedione (2, 5-HD), and explore the molecular mechanism of n-hexane neuropathy.
METHODSMale Wistar rats were administered at a dosage of 400 mg/kg/day 2, 5-HD for 2, 4 and 8 weeks respectively. HD-induced neurological defects were detected and quantified using gait score, and the relative lev-els of NF-H, NF-M, and NF-L in spinal cords of rats were determined by Western Blotting.
RESULTSExposure to 2, 5-HD produced progressive gait abnormalities, which suggested that the rat model of 2, 5-HD-induced neurotoxicity was established successfully. Western-Blotting results showed that NFs content in spinal cord demonstrated a progressive decline as the intoxication continued. In the supernatant fraction, compared to the controls, NF-H con-tent decreased by 15.7%, 57.0%, and 58.0% respectively after 2, 4, and 8-week treatment with 2, 5-HD (P < 0.01); accordingly, NF-M decreased by 36.0%, 61.3%, and 65.2% respectively (P < 0.01); NF-L decreased by 20.8%, 43.9%, and 44.3% respectively (P < 0.01). In the pellet fraction, the contents of NF-H in groups of 4 and 8 weeks' exposure to HD decreased by 35.6% and 43.2%, respectively (P < 0.01), and those of NF-L decreased by 26.4% and 42.1%, respectively (P < 0.01) when compared to the control. Further-more, NF-M contents in groups of 2, 4 and 8 weeks' exposure decreased by 23.3%, 33.9%, and 63.7% respectively (P < 0.01). The NFs level in spinal cords was highly correlated with gait abnormality of treated rats as the intoxication went on. Multiple correlation coefficients of NF-H, NF-M, and NF-L content with gait score of HD-treated rat were 0.8912, 0.9282 and 0.8981 (P < 0.01) respectively.
CONCLUSIONThe declines of NFs are high-ly related to neurobehavioral abnormality of 2, 5-HD-treated animals, and involved in the development of n-hexane neuropathy.
Animals ; Disease Models, Animal ; Gait ; drug effects ; Hexanones ; toxicity ; Male ; Neurofilament Proteins ; metabolism ; Rats ; Rats, Wistar ; Spinal Cord ; drug effects ; metabolism
9.In vivo and in vitro determination of changes of electrophysiology in rats with allyl chloride poisoning.
Qing-shan WANG ; Ying-jian ZHU ; Li-ping ZHANG ; Xin GUO ; Ke-qin XIE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2007;25(8):460-464
OBJECTIVETo compare the results of in vivo and in vitro in determination of the changes of allyl chloride (AC)-induced electrophysiology in rats sciatic nerve.
METHODSNinety male Wistar rats weighted 180 approximately 220 g were divided randomly into two groups, i.e. experimental group (n=40) and control group (n=50). The rats in experimental group were treated with AC dissolved in corn oil (200 mg/kg ip 3 days/week) by gavage for 12 weeks. Electrophysiological indexes of each group were determined on 3, 6, 9 and 12 weeks of AC intoxication. The indexes included measurements of sciatic nerve conduct velocity (NCV), compound action potential amplitude (CAPA), potential latency (PL), time course (TC), threshold potential (TP) and max stimulate potential (MSP).
RESULTSCompared to the corresponding time-matched control rats, on 6, 9 and 12 weeks of AC intoxication, NCV were decreased by 23.6%, 40.4% and 48.6% (P<0.05, P<0.01) in vivo, while in vitro it was decreased by 15.4% (P<0.05) on 12 week, CAPA were reduced by 31.7% in vivo, while in vitro it was reduced by 31.7%, 38.9% and 58.9% (P<0.05, P<0.01), respectively, PL were prolonged 22.6% and 40.7% (P<0.01) on 9, 12 weeks in vivo, while in vitro it was prolonged 8.0% (P<0.05), TC were increased 22.5%, 34.6% and 47.5% (P<0.01) in vivo, while in vitro it was increased 11.6%, 20.0% (P>0.05) and 19.5% (P<0.01), respectively, TP were elevated 12.1% (P>0.05), 32.3% and 40.0% (P<0.05) in vivo, while in vitro it was elevated 16.4% (P>0.05), 29.2% and 35.6% (P<0.05), respectively, MSP were increased 40.5% (P>0.05), 69.0% and 86.5% (P<0.01) in vivo, while in vitro it was increased 29.7% (P>0.05), 52.0% and 61.9% (P<0.01), respectively.
CONCLUSIONThe two methods of in vivo and in vitro showed that AC could significantly affect the electrophysiology of sciatic nerve, and the time-dependent changes occurred. The NCV is the most sensitive indicator in vivo to the early diagnosis of AC intoxication, while CAPA is the most sensitive indicator in vitro.
Action Potentials ; drug effects ; physiology ; Allyl Compounds ; poisoning ; Animals ; Disease Models, Animal ; In Vitro Techniques ; Male ; Neural Conduction ; drug effects ; physiology ; Random Allocation ; Rats ; Rats, Wistar ; Sciatic Nerve ; physiopathology
10.Effect of carbon disulfide exposure at different phases on the embryonic development in mid-pregnancy of female mice.
Zhi-ping WANG ; Ke-qin XIE ; Hui-qing LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2005;23(2):139-141
OBJECTIVETo explore the sensitive point of embryotoxicity of carbon disulphide on female mice.
METHODSAt the phases of follicle developing, implanting and post-implantation of blastocyst, female mice were injected intraperitoneally with carbon disulphide 631.4 mg/kg per day for three days while controls with plant oil. All indexes were detected at the fourteenth day of pregnancy.
RESULTS(1) In follicle developing CS(2) exposed group, the weight of embryos fossa [(1.23 +/- 0.36) g was 41% less than that in controls [(2.08 +/- 0.48) g, P = 0.000], and in implanting CS(2) exposed group the weight of embryo fossa, and embryos [(1.27 +/- 0.97) g, and (0.12 +/- 0.09) g respectively] were 39% and 37% less than those in controls [(2.08 +/- 0.48), (0.19 +/- 0.06) g, P = 0.068, P = 0.045]; (2) In both follicle developing and implanting CS(2) exposed group, the weights of uterus and placenta were also less than those in controls (P < 0.01). (3) In post-implantation CS(2) exposed group, the above parameters were not significantly different from those in controls.
CONCLUSIONFollicle developing phase as well as implanting of blastocyst may be sensitive point for embryotoxicity induced by carbon disulphide.
Animals ; Carbon Disulfide ; toxicity ; Embryo Implantation ; drug effects ; Embryonic Development ; drug effects ; Female ; Male ; Mice ; Mice, Inbred Strains ; Pregnancy ; Teratogens ; toxicity