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Abdominal radical trachelectomy was usually utilized in ⅠB1 cervical cancer patients whose tumor ≥2 cm since the procedure removed more parametrial tissue than the vaginal approach.We raised selection criteria for ART in such patients in 2011. In this article,we will further discuss issues regarding fertility-sparing surgery in ⅠB1 cervical cancer with tumor ≥2 cm.
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Purpose To analyze the Wnt/β-catenin signaling pathway related TCF/LEF binding sites in human mesothelin gene and to identify the core promoter region of the gene in the ovarian cancer cells. Methods The possible TCF/LEF transcription factor binding sites in the promoter region of human meosothelin gene were analyzed by bioinformatics method. The 1764 bp promoter sequence near the 5'end of the human mesothelin gene were cloned. The fragments was truncated differently at the 5' end and cloned into p GL3-basic report gene vector and transfected into human ovarian cancer cell lines SKOV-3 and 3-AO. The activity of diffetent promoter fragmentis was detected by double luciferase reporter gene system. Results There were multiple potential TCF/LEF binding sites in the promoter region of the human mesothelin gene. Three fragments of mesothelin gene promoter region-1456-+ 308、-164-+ 308、+ 47-+ 308 were cloned and amplified successfully, the p GL3 vector was constructed by sequencing. After transfection of SKOV-3 and 3-AO cells, the double luciferase reporter gene system showed that the-1456-+ 308 fragments and-164-+ 308 framents had high promoting activity in both cell lines, and the activity of + 47-+308 fragments was significantly lower than that of the former two cell lines (P<0.01). Conclusion The-164-+ 47 sequence containing TCF/LEF transcription factor binding site is the core promoter region of mesothelin gene in ovarian cancer, which lays a foundation for further study on the regulation mechanism of mesothelin gene expression in ovarian cancer.
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Objective To summarize the key points of forensic medicine appraisal by retrospectively analyzing the scene.Methods The data of 193 homicide cases in a district of Weinan from 2007 to 2016 were collected and analyzed.Results The majority of the criminal suspect and victims was male,aged 21-40 years old,local,farmers and primary school educational level.Infringement was the majority criminal motivation (52.8％).The most common injuries tools were sharp instruments (63.8 ％);multiple parts damage accounted for 32.1 ％;mechanical injury induced hemorrhagic shock and craniocerebral injury were the most common causes of death in these cases (87.0％).Conclusions The demographic characteristics of suspects,criminal motivation,injury instruments,distribution of injuries and cause of death in the district of Weinan showed similar general characteristics,which could provide experience and reference for detecting homicide cases in the future.
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Due to the advantages in genetic manipulation, mice have become one of the most commonly used mammalian models for the study of mechanisms underlying myopia development. However, the vast majority of laboratory mouse strains are incapable of synthesizing melatonin, a neurohormone that may play an important role in myopia generation in humans. The present study investigated refractive development profiles in the CBA/CaJ mouse, a strain proficient in melatonin, and determined whether and how its refractive development could be affected by form-deprivation. Eccentric infrared photoretinoscopy revealed that this animal could be stably refracted, and the refractive error underwent developmental changes, which increased with age in the hyperopic direction and eventually got stable approximately 9 weeks after birth. The absolute values of refractive error in CBA/CaJ mice were larger than those of age-matched C57BL/6 mice, whereas the time points when refractive error reached steady state were similar between the two strains. Five weeks of form-deprivation applied to 3-week-old CBA/CaJ mice by translucent occluder wear caused a significant myopic shift in refractive error, indicating that this strain could be adequately used as a myopia model.
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Animals , Mice , Disease Models, Animal , Eye , Mice, Inbred C57BL , Mice, Inbred CBA , Myopia , Refraction, Ocular , Sensory DeprivationABSTRACT
Objective: To investigate the influence of maximum-tolerated dose chemotherapy with cisplatin (MTD-DDP) and low-dose metronomic chemotherapy with DDP (LDM-DDP) on human ovarian carcinoma xenograft tumor in nude mice and the expression of aldehyde dehydrogenase 1 (ALDH1) in xenograft tumor. Methods: Murine subcutaneous xenograft tumors of human epithelial ovarian cancer SKOV3 cells were established in nude mice and treated with MTD-DDP and LDM-DDP, respectively. The growth of xenograft tumor was observed. The xenograft tumor primary cells were established. ALDH1-positive cells were obtained by fluorescence-activated cell sorting. The abilities of colony formation and tumorigenicity of ALDH1-positive cells in vitro and in vivo were detected by colony formation assay and tumor formation in nude mice, respectively. The expression levels of Ki-67, breast cancer resistant protein (BCRP) and CD133 in ALDH1-positive cells were detected by Western blotting. Results: The growth of xenograft tumor in nude mice was inhibited by MTD-DDP and LDMDDP, and this inhibition effect was significant in LDM-DDP group (P < 0.001). The proportion of ALDH1-positive cells in xenograft tumor primary cells was lower in MTD-DDP group and higher in LDM-DDP group as compared with that in the control group (0.9% NaCl solution) (both P < 0.001). The abilities of colony formation and tumorigenicity of ALDH1-positive cells were higher than those of ALDH1-negative cells (both P < 0.001). The expression level of Ki-67 was lower but the expression levels of BCRP and CD133 were higher in ALDH1-positive cells than those in ALDH1-negative cells (all P < 0.001). Conclusion: LDM-DDP can effectively inhibit the growth of human ovarian carcinoma xenograft tumors in nude mice and reduce the proportion of ALDH1-positive cells.
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Objective: To investigate the impact of low-dose metronomic chemotherapy (LDM) and maximum tolerated dose chemotherapy (MTD) with cisplatin on BALB/c nude mice bearing human ovarian cancer and the xenograft tumors of SKOV3 cells, and to explore the influence of these two chemotherapeutic protocols on expression levels of breast cancer resistant protein (BCRP), lung resistance protein (LRP) and tumor stem cell marker CD133. Methods: The human ovarian cancer SKOV3 cells were used to establish BALB/c nude mice model bearing subcutaneous tumor xenografts. Thirty-six mice were randomly divided into three groups: control group (n = 12), LDM group (n = 12) and MTD group (n = 12). The general situation of nude mice and the growth of tumor xenografts were observed during chemotherapy, and the tumor inhibition rate was calculated. The expression levels of BCRP, LRP and CD133 proteins in tumor xenografts in nude mice were detected by Western blotting. The small metastatic lesions in internal organs and the pathological feature of tumor xenografts were observed by hematoxylin-eosin (HE) staining. Results: The volume of tumor xenografts of MTD and LDM groups were significantly smaller than that of the control group (P < 0.05), while the tumor volume of LDM group was significantly smaller than that of MTD group. The tumor inhibition rates of MTD group and LDM group were 27.00% and 54.29%, respectively. The body weight and food-intake of mice in MTD group and LDM group were both significantly lower than those in the control group (all P < 0.05). As compared to LDM group and the control group, white blood cell (WBC) count in MTD group was lower (P < 0.05). Hepatocellular fatty degeneration was observed in two mice in MTD group. The expression levels of BCRP, LRP and CD133 proteins in tumor xenografts in MTD group and LDM group were significantly higher than those in the control group (all P < 0.05), while the BCRP and CD133 protein expression levels in LDM group were lower than those in MTD group. Conclusion: LDM can lead to greater tumor growth inhibition and less bone marrow suppression as compared with MTD, at the same total cumulative dose. Tumor cells with partial tumor stem cell properties and high expression levels of drug resistance-related proteins were easier to be enriched in MTD model. LDM is a new potential therapeutic mode which may reduce tumor recurrence and increase the sensitivity to chemotherapy.
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Objective: To investigate the relationship between the autophagy and the sensitivity to DDP (cisplatin) in epithelial ovarian cancer SKOV3 and SKOV3/DDP cells. Methods: The formation of autophagosomes in SKOV3 and SKOV3/DDP cells was observed under a TEM (transmission electron microscope). MTT assay was used to screen the dose of nontoxic reaction of autophagy inhibitor 3-MA (3-methyladenine) and to examine the sensitivity to DDP in SKOV3 and SKOV3/DDP cells after treatment with the combination of DDP and 3-MA. The apoptotic rate and the expressions of autophagic marker protein LC3 (microtubule-associated protein 1 light chain 3) and autophagy-related protein Beclin1 in SKOV3 and SKOV3/DDP cells after treatment with DDP and 3-MA alone and the combination of DDP and 3-MA were detected by flow cytometry and Western blotting, respectively. Results: Much more autophagosomes in SKOV3/DDP cells were observed as compared with that in SKOV3 cells. As compared with DDP treatment alone, when DDP was used in combination with 3-MA, the IC50 (half inhibitory concentration) value of DDP for SKOV3/DDP cells was decreased from (17.71±0.15) μg/mL to (8.85±0.11) μg/mL (P < 0.05), and the apoptotic rate was increased from (6.93±0.94) % to (14.82±0.85) % (P < 0.05), as well as the expression levels of LC3 and Beclin1 proteins in SKOV3/DDP cells were significantly decreased (P < 0.05), but there were no significant changes could be observed in SKOV3 cells. Conclusion: The increased activity of autophagy is related to DDP-resistance of ovarian cancer cells. The inhibition of autophagy may enhance the sensitivity to DDP in SKOV3/DDP cells. Copyright © 2013 by TUMOR.
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<p><b>OBJECTIVE</b>Metabonomic analysis has been increasingly used to monitor metabolic abnormalities in cells and their micro-environment in order to detect the biomarkers recently. This study evaluated the feasibility of applying 1H-nuclear magnetic resonance (1H-NMR) based metabonomic method to detect the differences of the early development of cleft palate in the plasma from control group and experimental group.</p><p><b>METHODS</b>Pregnant mice (inbred C57BL/6J strain) with vitamin B12 injected only were assigned as the control group, pregnant mice with excessive Dex, injected after vitamin B12 as the experimental group, each group includes 12 mice. And the effect of B12 to rate of cleft palate was observed. The technology of nuclear magnetic resonance (NMR) was used to detect the endogenous small molecule metabolites. Finally, changes of metabolites ingredients were ascertained by using the method of principal component analysis (PCA).</p><p><b>RESULTS</b>There was significant difference in PCA scores plot between the two groups according to whether cleft palate occurred.</p><p><b>CONCLUSION</b>The 1H-NMR based metabonomic approach might be used as a feasible and efficient method for a deep exploration of the pathogenesis of cleft lip and palate and an early exploration of the mechanism of vitamin B12.</p>
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Animals , Female , Mice , Pregnancy , Cleft Lip , Cleft Palate , Dexamethasone , Magnetic Resonance Imaging , Magnetic Resonance Spectroscopy , Metabolomics , Mice, Inbred C57BL , Principal Component Analysis , Vitamin B 12ABSTRACT
<p><b>OBJECTIVE</b>To study possible impairment mechanisms of learning and memory abilities from unsaturated fatty acids in hippocampus of mice exposed to lead.</p><p><b>METHODS</b>Forty-eight healthy mice were divided into 4 groups: low dose (0.625 g/L), middle dose (1.250 g/L) and high dose (2.500 g/L) of lead solution in diet and control group (distilled water). The mice in treatment groups were fed with lead solution every day while the mice in control group were fed with distilled water for 50 days. After learning and memory abilities were measured, the mice were killed and contents of oleic acid (C18:1), linoleic acid (C18:2), linolenic acid (C18:3), arachidonic acid (AA,C20:4), eicosapentaenoic acid (EPA,C20:5) and docosahexaenoic acid (DHA, C22:6 ) in hippocampus of mice were determined by high performance liquid chromatography (HPLC).</p><p><b>RESULTS</b>(1) In the four training days, the mice treated with lead in the middle dose group and high dose group significantly increased the escape latencies compared with the mice treated with distilled water (P<0.05), and on the 4th day, the low dosage mice's escape latencies were delayed (P<0.05). The escape latencies of the 1st, 2nd, 3rd and 4th day had significantly positive linear relation with lead dose. Their relative coefficient in turn is r=0.973, 0.985, 0.929 and 0.936, indicating that lead harmed spatial memory of mice in Morris water maze (MWM). (2) The contents of C18:2 and AA were obviously enhanced in hippocampus of middle and high dosage (P<0.05); while there was evident decrease in the contents of C18:3, EPA and DHA (P<0.05); the content of C18:1 was decreased significantly in high dosage group (P<0.01). The mice's escape latencies had significantly negative linear relation with contents of C18:1, C18:3, EPA and DHA, while there was positive linear relation significantly with contents of C18:2 and AA. Their relative coefficient in turn was r=-0.901, -0.914, -0.893, -0.855, 0.936, 0.727.</p><p><b>CONCLUSION</b>Lead interferes with the metabolism of hippocampus fatty acids and affects membrane function in hippocampus of mice, which might contribute to change of the synthesis, metabolism and release of central neurotransmitter and decrease of the learning and memory abilities.</p>
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Animals , Female , Male , Mice , Fatty Acids, Unsaturated , Metabolism , Hippocampus , Metabolism , Lead , Toxicity , Maze Learning , Mice, Inbred ICRABSTRACT
<p><b>OBJECTIVE</b>To understand the adverse and curative effects of film coated praziquantel for treatment of schistosomiasis japonica.</p><p><b>METHODS</b>A questionnaire survey was conducted among 6 to 65 years old inhabitants from epidemic areas of schistosomiasis japonica in Jiangxi, Anhui and Hubei provinces, while indirect haemagglutination assay (IHA) was also administrated. The Serological positives were tested by Kato-Katz technique. A total of 509 habitats of serological positives and healthy persons were given praziquantel. 460 targets with no relative symptoms were followed up for adverse effect of drugs in one month and efficacy evaluation of praziquantel was carried out in 104 parasitological positives 3 months post-treatment.</p><p><b>RESULTS</b>The 84.7% (144/170) individuals thought film coated tablets were no smell or only with little smell while 92.9%(315/339) targets in control group fell that tablets smelled bad. The total side effect rates of film coated praziquantel group and control group in 1-2 hours post-treatment were 20.30% (27/133) and 83.18% (272/327) respectively. There was significantly different between two groups (chi(2) = 164.316, P < 0.05). The adverse effect rates of film coated praziquantel group in neuromuscular system, digestive system and cardiovascular system were 15.79% (21/133), 9.77% (13/133), 2.26% (3/133) individually which were significantly lower than those of control groups with 81.65% (267/327), 49.24% (161/327), 12.84% (42/327) in corresponding systems (chi(2)(neuromuscular system) = 175.188, chi(2)(digestive system) = 62.601, chi(2)(cardiovascular system) = 12.010, P < 0.05). And the adverse effect rate of allergic reaction of film coated group was no significant difference with control group (2.26%(3/133) versus 0.92% (3/327), chi(2) = 1.315, P = 0.235). One day after treatment, the adverse effect rate of film coated tablets of praziquantel decreased to 3.01% (4/133), significantly lower than that of control group (P < 0.05). There was no significant difference after 2 weeks treatment while the adverse effect rates decreased to 0.75%(1/133), 0.61% (2/327) in film coated praziquantel group and control group respectively (chi(2) = 0.029, P = 0.642). The curative rates 3 months post treatment were 84.91% (45/51) in film coated tablets of praziquantel group and 82.35% (42/53) in control group without significant difference (chi(2) = 1.536, P = 0.215).</p><p><b>CONCLUSION</b>The smell and adverse effects of film coated tablets of praziquantel were decreased significantly and its efficacy for treatment of schistosomiasis japonica was equal to the control tablets. The film coated tablets of praziquantel might be applied in field widely after a further verification.</p>
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Adolescent , Adult , Aged , Child , Humans , Male , Middle Aged , Young Adult , China , Praziquantel , Therapeutic Uses , Schistosomiasis japonica , Drug Therapy , Tablets , Treatment OutcomeABSTRACT
<p><b>BACKGROUND</b>Significant progress has taken place over the past 50 years in the control of schistosomiasis japonica in China. However, the available data suggested that schistosomiasis has re-emerged shortly after the World Bank Loan Project which was conducted from 1992 to 2001. The national control program with a revised strategy to control schistosomiasis by using integrated measures has been implemented since 2005. In this study, we aimed to evaluate the effect of the national program on schistosomiasis control from 2005 to 2008.</p><p><b>METHODS</b>A retrospective study was carried out to analyze the epidemic patterns of acute infections with Schistosoma japonicum (S. japonicum), based on the number of acute cases annually collected from the web-based national communicable diseases reporting system from 2005 to 2008.</p><p><b>RESULTS</b>A total of 564, 207, 83 and 57 acute cases infected with S. japonicum were reported nationwide in 2005, 2006, 2007 and 2008, respectively, with an average annual reduction rate of 46.35% during last four years. Six outbreaks of acute infection with S. japonicum were reported in 2005 but none in the period of 2006 to 2008. All acute cases that were reported mainly came from the lake regions and became infected during the higher risk periods from the 27th to 43rd weeks of the year. Most of these cases are students (44.87%), farmers (31.51%) and fishermen (7.79%) who got the infection by water contact mainly through swimming (41.49%) and production activities (40.25%). With time, the proportion of imported cases among all acute cases increased due to more frequent movement of people that has occurred with a more mobile population.</p><p><b>CONCLUSIONS</b>The national control program on schistosomiasis aliened with the revised control strategy has been effectively brought into effect. However, there is still a significant risk of infection among students, farmers and fishermen living in the lake regions. Therefore, it is important to strengthen control measures among risk populations in the high risk areas of transmission, or the lake regions.</p>
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Animals , Humans , Acute Disease , Epidemiology , China , Epidemiology , Retrospective Studies , Schistosoma japonicum , Virulence , Schistosomiasis , EpidemiologyABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of survivin antisense oligonucleotides (ASODN) on survivin and Smac gene expression in ovarian cancer SKOV3 cells and explore the role of survivin and Smac genes in ASODN-induced ovarian cancer cell cycle alteration and apoptosis and its molecular mechanisms.</p><p><b>METHODS</b>Survivin ASODN was introduced via Lipofectamine(TM)2000 into SKOV3 cells, whose growth activity was detected subsequently with MTT assay. The apoptosis index, cell cycle and changes in survivn and Smac protein expression were determined using flow cytometry. The changes in survivn and Smac mRNA expression were detected by RT-PCR.</p><p><b>RESULTS</b>In comparison with the control cells, cells transfected with difference concentrations of survivin ASODN exhibited significantly inhibited growth, and 48 h after the transfection, the IC(50) was about 600 nmol/L. After a 48-hour transfection of the cells with 600 nmol/L ASODN, the apoptosis index significantly decreased (t=6.3671, P<0.05), and the cell percentage in (1)/G(0) phase increased (t=10.3832, P<0.01), resulting also in significantly down-regulated survivin mRNA and protein expressions (t=3.5031, P<0.05; t=7.8146, P<0.01) and up-regulated Smac mRNA and protein expressions (t= 2.8011, P<0.05; t= 11.3917, P<0.01).</p><p><b>CONCLUSION</b>ASODN against survivin can induce apoptosis of ovarian cancer cell line SKOV3 and results cell growth arrest in G(1)/G(0) phase by up-regulating Smac and down-regulating survivin expression. Survivin and Smac are closely correlated in their action on SKOV3 cell cycle and apoptosis, which is one of the important mechanisms of ovarian cancer development.</p>
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Female , Humans , Apoptosis , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Inhibitor of Apoptosis Proteins , Intracellular Signaling Peptides and Proteins , Genetics , Metabolism , Microtubule-Associated Proteins , Genetics , Metabolism , Mitochondrial Proteins , Genetics , Metabolism , Oligonucleotides, Antisense , Genetics , Metabolism , Ovarian Neoplasms , Genetics , MetabolismABSTRACT
<p><b>BACKGROUND</b>In a previous study, we have verified that CXCR4 expression is correlated with tumor aggressive progression and poor prognosis in patients with epithelial ovarian cancer. The aim of this study was to explore the effect of CXCL12-CXCR4 axis on the metastasis of human ovarian cancer.</p><p><b>METHODS</b>The expressions of CXCR4 and CXCL12 mRNA and protein in human ovarian cancer cell line CAOV-3 was detected by RT-PCR and immunocytochemistry. Methythiazolyltetrazolium (MTT) was used to analyze the effect of different concentrations of CXCL12 on the proliferation of CAOV-3 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of CXCL12 and ascites on the migration and invasion of CAOV-3 cells. The expressions of integrin beta(1) and vascular endothelial growth factor-C (VEGF-C) mRNA were detected by RT-PCR. Data were analyzed using ANOVA by SAS 6.12.</p><p><b>RESULTS</b>Under serum-free suboptimal culture conditions, CXCL12 (100 ng/ml) significantly enhanced the proliferation of CAOV-3 cells compared with the control and 10 ng/ml CXCL12 groups (0.428 +/- 0.051 vs. 0.325 +/- 0.045 and 0.328 +/- 0.039, P < 0.05). This enhancing effect of CXCL12 was significantly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml CXCR4 antagonist AMD3100. However, 10 microg/ml neutralizing CXCR4 antibody could not inhibit cell proliferation without CXCL12. The levels of migration and invasion of the CAOV-3 cells treated with 100 ng/ml CXCL12 were significantly higher than those in the control (migration: 523.3 +/- 25.2 vs 108.0 +/- 7.2; invasion: 39.3 +/- 4.0 vs. 4.0 +/- 1.0). The enhancing effect of CXCL12 on cell migration and invasion increased with the concentration of CXCL12 (100 ng/ml vs10 ng/ml: migration, 523.3 +/- 25.2 vs 211.7 +/- 24.7; invasion, 39.3 +/- 4.0 vs 15.7 +/- 3.1, P < 0.05), and was strongly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml AMD3100. The number of migrated and invading cells in the CAOV-3 added with ascites was significantly higher than those in the 100 ng/ml CXCL12 group (migration: 706.6 +/- 30.6 vs 523.3 +/- 25.2, invasion: 61.7 +/- 7.6 vs 39.3 +/- 4.0, P < 0.05). The level of integrin beta(1) mRNA was greatly increased at 3 hours after being treated with CXCL12 (0.53 +/- 0.10 vs. 1.53 +/- 0.16, P < 0.05), and VEGF-C mRNA displayed significant augment at 24 hours after being treated with CXCL12 (0.52 +/- 0.09 vs 1.11 +/- 0.15, P < 0.05).</p><p><b>CONCLUSIONS</b>CXCL12 and its receptor CXCR4 can promote the proliferation, migration, invasion of ovarian cancer cell line CAOV-3 and enhance its secretion of integrin beta(1) and VEGF-C. These effects can be inhibited by neutralizing CXCR4 antibody or AMD3100. CXCL12-CXCR4 axis plays an important role in ovarian cancer growth and metastasis.</p>
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Female , Humans , Cell Line, Tumor , Cell Movement , Cell Proliferation , Chemokine CXCL12 , Chemokines, CXC , Physiology , Neoplasm Invasiveness , Neoplasm Metastasis , Ovarian Neoplasms , Pathology , Receptors, CXCR4 , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate effect of lysophosphatidic acid (LPA) on growth and metastasis of intraperitonally transplanted tumor in nude mice.</p><p><b>METHODS</b>Sixteen nude mice randomly divided into control group and LPA treatment group, in which 6x10(6) untreated and LPA-treated human ovarian cancer cells skov3 were respectively transplanted intraperitoneally. After death of the nude mice without interventions, the weight of the transplanted tumors and number of metastatic tumors were recorded.</p><p><b>RESULTS</b>The rate of tumor formation was 100%in both groups, but the treatment group showed more rapid tumor growth, greater tumor weight and more numerous metastatic tumors than the control group (P<0.01).</p><p><b>CONCLUSION</b>LPA simulates the growth and promotes invasiveness of transplanted ovarian cancer cells in the peritoneal cavity of nude mie.</p>
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Animals , Female , Mice , Lysophospholipids , Toxicity , Mice, Nude , Neoplasm Invasiveness , Neoplasm Transplantation , Ovarian Neoplasms , Pathology , Peritoneal Neoplasms , Random Allocation , Transplantation, Heterologous , Tumor BurdenABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of N-methyl-D-asparate (NMDA) receptor subunit proteins after administration of different doses of pentylenetetrazol (PTZ).</p><p><b>METHODS</b>After ip injection of a subconvulsant (35 mg/kg) and convulsant (50 mg/kg) dose of PTZ, the rats were decapitated at different time points. The levels of cortical NR1 NR2A and NR2B subunit proteins were detected by immunoblotting.</p><p><b>RESULT</b>35 mg/kg PTZ and 50 mg/kg PTZ elicited different behavioral changes (P<0.001). The NR2A subunit in the cortex significantly increased 1 h after PTZ injection (P<0.05). For the 50 mg/kg group, both the NR2A and NR2B subunits proteins increased at 1 h in the cortex and then decreased; the protein levels returned to normal after 48 h. However, NR1 subunit had no changes.</p><p><b>CONCLUSION</b>The NR2 subunit is involved in PTZ-induced seizure.</p>
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Animals , Male , Rats , Cerebral Cortex , Chemistry , Dose-Response Relationship, Drug , Pentylenetetrazole , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate , Seizures , MetabolismABSTRACT
Objective To explore the effect of chemokine CXCL12 and its receptor CXCR4 on proliferation,migration and invasion of epithelial ovarian cancer cells.Methods CXCR4 and CXCL12 mRNA and protein expression of human ovarian cancer cell line CAOV3 was detected by RT-PCR and immunocytochemistry.Integrin ?1 and vascular endothelial growth factor-C(VEGF-C)mRNA expression were detected in CAOV3 cells stimulated by CXCL12.The CAOV3 cells were divided into 6 groups:control group(un-stimulated),experimental group 1(stimulated by 100 ng/ml CXCL12),experimental group 2 (stimulated by 10 ng/ml CXCL12),experimental group 3(100 ng/ml CXCL12 and 10 ?g/ml neutralizing CXCR4 antibody),experimental group 4(100 ng/ml CXCL12 and 1 ?g/ml CXCR4 antagonist AMD3100),experimental group 5(10 ?g/ml neutralizing CXCR4 antibody or ascites).Methyl thiazolyl tetrazolium(MTT)was used to analyze the effects of different concentrations of CXCL12 on CAOV3 cell proliferation.Transwell invasion chamber and reconstructed basement membrane(Matrigel)were used to evaluate effect of various concentrations of CXCL12 and ascites on CAOV3 cell migration and invasion. Results CAOV3 cells expressed CXCR4 mRNA(0.70?0.10)and protein,but did not express CXCL12 mRNA or protein.Immunostaining of CXCR4 was mainly located in cytoplasm.CXCR4 mRNA was up- regulated after 100 ng/ml CXCL12 stimulation(1.24?0.14;t=-7.1088,P=0.0021).Integrin ?1 mRNA was greatly increased at 3 hours by stimulation of 100 ng/ml CXCL12(before and after stimulation 0.53?0.10,1.53?0.16;P0.05).Experimental group 1 stimulated the migration and invasion of CAOV3 cells in chemotaxis assay compared with control group and experimental group 2(number of cell migration respectively 523.3?25.2,108.0?7.2,211.7 ?24.7,number of cell invasion respectively 39.3?4.0,4.0?1.0,15.7?3.1;P