Objective: To investigate the effect of taurine on learning and memory impairment, cytokines secretion in rats intrahippocampally injected with ?-amyloid (A?) 1-40. Methods: SD rats were randomly divided into control group, A? injected group, taurine (0.3g/kg?d, 0.6g/kg?d) groups. The rats were fed with taurine for 7 days, and then subjected to bilateral intrahippocampus injection of A?1-40 or vehicle. Two weeks later, all rats performed Morris water maze test. The contents of IL-6, TNF-? were checked by way of radio-immunity assay for hippocampus samples. Results: Compared with A?model group, the escape latency and distance were significantly reduced in taurine (0.6g/kg?d) group; the ratio of swimming distance in the target quadrant to that in the whole pool of the probe trial; the content of cytokines of IL-6 and TNF-?in hippocampus were reduced significantly. Conclusion: Taurine can effectively attenuate the cognitive dysfunction caused by A?1-40 in rats. The reduced cytokines content in hippocampus might contribute to this effect.

Objective To investigate the relationship between serum follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and clinicopathological features and prognosis of serous ovarian cancer retrospectively. Methods A total of 73 patients with serous ovarian cancer treated in the Department of Obstetrics and Gynecology of Tianjin Medical University General Hospital from January 2000 to December 2015 were included in this study. The relationship between serum FSH, LH, PRL and clinicopathological features was analyzed by Mann-Whitney U method. Kaplan-Meier (K-M) method was used to analyze survival rates of patients with different clinical features. Multivariate Cox proportional hazards regression analysis was used to analyze prognostic factors of serous ovarian cancer patients. Results The mean concentrations of serum FSH and LH were significantly higher in the>50 year-old group than those in the<50 year-old group (P<0.05). The mean concentrations of FSH and LH were significantly higher in menopause group than those in non-menopause group (P<0.05). There were no significant differences in serum levels of FSH and LH in patients with other different clinicopathological features (P>0.05). There was no significant correlation between serum PRL concentration and clinicopathological features (P>0.05). Analysis results showed that poor prognosis of patients was related with high serum levels of FSH (>40.13 IU/L), PRL (>14.96 μg/L) and FIGO stage (Ⅲ+Ⅳ) (P<0.05). There was no significant correlation between serum LH concentration and prognosis (P>0.05). COX regression analysis showed that the serum PRL>14.96 μg/L was risk factor for prognosis of serous ovarian cancer [HR(95%CI): 3.530(1.180-10.557),P=0.024]. Conclusion The serum levels of FSH and LH are significantly increased in postmenopausal women than those in menopause women. The serum level of PRL is correlated with the prognosis of serous ovarian cancer.

OBJECTIVETo study the role of Ezh2 in the all-trans retinoic acid RA induced P19 neural differentiation.

METHODSThe expression of Ngn1 in the RA induced P19 cells was detected at the mRNA and protein levels using real time RT-PCR and Western blot assays. The binding of Ezh2 and H3K27me3 on the Ngn 1 promoter was analyzed using chromatin immunoprecipitation assay.

RESULTIn the RA induced P19 cells, the recruitment of Ezh2 and its methylated substrate H3K27me3 on the promoter of Ngn 1 gene elevated in the first 2 days, and then declined rapidly, followed by the initiation of neuronal differentiation.

CONCLUSIONSEzh2 produces a repressive histone mark H3K27me3 in the early stage of RA induced P12 cells. By avoiding the premature expression of Ngn1 gene, Ezh2 can ensure the normal differentiation of P19 cells.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Differentiation ; drug effects ; physiology ; Enhancer of Zeste Homolog 2 Protein ; Histone-Lysine N-Methyltransferase ; genetics ; metabolism ; Histones ; metabolism ; Mice ; Nerve Tissue Proteins ; genetics ; metabolism ; Neurons ; cytology ; drug effects ; metabolism ; Polycomb Repressive Complex 2 ; Tretinoin ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo investigate the effect of the new Traditional Chinese Compound Shenwu Capsule on the damage of lymphocyte DNA and lipid peroxidation in peripheral blood of rats induced by beta-amyloid injection.

METHODThe animal model was made by injection of beta-amyloid25-35 into hippocampus of rats. DNA damage of lymphocytes was measured by the single cell gel electrophoresis (SCGE) combined with the laser scanning confocal microscopy (LSCM). The content of malondialdehyde (MDA) was determined by TBA assay.

RESULTShenwu Capsule decreased the rate of the comet-like cell, comet-like cell lengh (TCL), and tail moment (TM) of lymphocytes in peripheral blood of model rats. Shenwu Capsule also declined the MDA content in serum of Abeta model rats.

CONCLUSIONShenwu Capsule has protective effect on peripheral blood lymphocyte DNA.

Amyloid beta-Peptides ; Animals ; Capsules ; DNA Damage ; drug effects ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Lymphocytes ; drug effects ; Male ; Neurodegenerative Diseases ; chemically induced ; pathology ; Neuroprotective Agents ; administration & dosage ; pharmacology ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Pueraria ; chemistry ; Rats ; Rats, Sprague-Dawley

Objective To establish a REDE-DHPLC method for detecting the EGFR and KRAS mutations in plasma DNA from tumor patients, and investigate its clinical significance. Methods Restriction endonucleases Mse Ⅰ , Msc Ⅰ , BstN Ⅰ and Bgl Ⅰ were used to digest the wild type fragments of exon 19,exon 21 of EGFR gene and coden 12, 13 of KRAS gene for enriching the mutation fragments, and REDE-DHPLC method was established to detect EGFR and KRAS mutations. The sensitivities of REDE-DHPLC and conventional DHPLC were analyzed by using a series of plasmids containing 50%, 10%, 5%, 1% and 0. 1% mutation genes. Then, Plasma samples and paraffin-embedded tissue samples of 120 NSCLC patients and 120 colorectal cancer patients were detected by REDE-DHPLC. Compared with conventional DHPLC and sequencing, the diagnostic efficiency of REDE-DHPLC method was evaluated by detecting the mutation status of 2 genes in plasma of NSCLC and colorectal cancer patients. Results The sensitivity values of REDE-DHPLC and conventional DHPLC for detecting mutations in 4 loci were 0. 1% and 1%respectively. Plasmid DNA containing 0.1% mutation gene was detected to be positive continually for 2 to 3 times by REDE-DHPLC. EGFR mutation rates of 120 plasma from NSCLC patients detected by REDE-DHPLC, conventional DHPLC and sequencing methods were 27. 5%, 16. 7% and 12.5% respectively, and KRAS mutation rates of 120 plasma from colorectal cancer patients were 38. 3%, 25. 8% and 16. 7%,respectively. The positive rates of EGFR and KRAS mutation detected by REDE-DHPLC were significantly higher than conventional DHPLC(x2 = 4. 092, 4. 301, all P ＜ 0. 05 ) and sequencing method (x2= 8. 438,14. 127,all P ＜ 0. 05 ). In comparison with conventional DHPLC, the sensitivities of REDE-DHPLC for detecting EGFR and KRAS mutation were 100% (20/20,31/31), the specificities were 87. 0% (87/100)and 83. 2% (74/89). In comparison with sequencing method, the sensitivities of REDE-DHPLC were 100%( 15/15,20/20), the specificities were 82.9% (87/105)and 74. 0% (74/100). The coincidence rate of the two methods for detecting EGFR and KRAS mutation were 89. 2% ( 107/120, Kappa = 0. 690, P ＜ 0. 05 ) and 87.5% ( 105/120, Kappa= 0. 718, P ＜ 0. 05 ). The Consistency of EGFR and KRAS mutation status in plasma and tissues detected by REDE-DHPLC were 91.7% (33/36, Kappa =0. 939,P ＜0. 05)and 90. 2 %(46/51, Kappa = 0. 914, P ＜ 0. 05 ), respectively. Conclusions The REDE-DHPLC method is highly sensitive and specific for detecting EGFR and KRAS mutations in plasma DNA from tumor patients. The results are easy to be interpreted without missing homozygous point mutation, which indicate that the detection of EGFR and KRAS mutations in plasma DNA by REDE-DHPLC could therefore extend to be usedin clinical laboratory.

OBJECTIVETo study the effects of exogenous ER beta on the growth of breast cancer MCF-7 cells under different treatment.

METHODSAn eukaryotic expression vector containing 1.6 kb of human entire coding sequence of ER beta (pCDNA3-ER beta) was transfected into human breast cancer MCF-7 cells using lipofectamine 2000. The biological activity of ER beta was detected with the luciferase reporter containing estrogen responsive element (ERE) and the expression of ER beta protein by Western blot. The growth properties of MCF-7, pCDNA 3-transfected MCF-7 and pCDNA 3-ER beta-transfected MCF-7 cells under different treatment, including E2 (17beta-estradiol) and 4-OHT (4-hydroxytamoxifen), were observed.

RESULTSA stronger activation of the reporter by ER beta in the presence of E2 was observed in the pCDNA 3-ER beta-transfected MCF-7 cells than in the pCDNA 3-transfected MCF-7 and in MCF-7 cells. Western blot analysis showed that the protein level of ER beta in the pCDNA 3-ER beta-transfected MCF-7 cells was markedly increased. Exogenous ER beta expression did not change the growth properties and the morphology of MCF-7 cells under normal condition. The pCDNA 3-ER beta-transfected MCF-7 cells proliferated at the same rate as naive cells in the presence of 4-OHT, whereas a strong inhibition of the proliferation of the pCDNA 3-ER beta-transfected MCF-7 cells in the presence of E2 was observed.

CONCLUSIONExogenous ER beta expression does not increase the resistance to 4-OHT, and a strong inhibition of the proliferation may occur in the presence of E2.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estradiol ; pharmacology ; Estrogen Antagonists ; pharmacology ; Estrogen Receptor beta ; genetics ; metabolism ; Female ; Humans ; Tamoxifen ; analogs & derivatives ; pharmacology ; Transfection

OBJECTIVETo provide more detailed information on the roles of lipid peroxidation in the pathogenesis of chronic pancreatic injuries in a pre-clinical rat model.

METHODSTotally 72 rats were divided into 6 groups (12 in each group) Rats in 5 experimental groups (n = 12) were fed with a high-fat diet (1% cholesterol, 10% lard, 0.3% sodium tauroglycocholate, 87.3% standard rodent chow as the control group) for 2, 4, 6, 10 and 16 weeks, respectively. Morphological studies in the pancreas tissue samples from rats were investigated by using various histological methods. Pancreatic stellate cells (PSCs) were identified by immunohistochemical staining for Desmin and α-smooth muscle actin (α-SMA). The expression of the lipid peroxidation was detected by immunostaining for 4-hydroxy-2-nonenal (4-HNE) and thromboxane A2 receptor (TxA2r). The co-localization of α-SMA and 4-HNE or α-SMA and TxA2r in PSCs was also analyzed in this study.

RESULTSPancreatic cells with positive staining for Desmin and α-SMA in HFD rats were distributed in a more extensive way when compared to that in the control group. The levels of pancreatic 4-HNE and TxA2r were increased in rats from HFD groups significantly. The co-localization of 4-HNE and TxA2r were also found within activated PSCs in both of groups.

CONCLUSIONThe results showed that a chronic HFD feeding may increase the lipid peroxidation process and collagen synthesis through a critical signaling pathway of activated PSCs following pancreatic injuries in rats.

Actins ; metabolism ; Aldehydes ; metabolism ; Animals ; Collagen ; biosynthesis ; Desmin ; metabolism ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Lipid Peroxidation ; Male ; Oxidative Stress ; Pancreas ; metabolism ; pathology ; Pancreatic Diseases ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Receptors, Thromboxane A2, Prostaglandin H2 ; metabolism

Objective: To explore the cumulative effects of unintentional injury among children in rural area, in order to provide information for early intervention of unintentional injury. Methods: Through multistage clustering sampling method, 2 109 primary caregivers of students from 8 rural primary and elementary schools of Heilongjiang Province were recruited. Strengths and Difficulties Questionnaire (SDQ), Injury Behavior Checklist (IBC), Perceptions of Risks and Hazards were used to collect as the risk factors, while Perceptions of Risks and Hazards (PSAPQ), Home Observation for Measurement of the Environment (HOME) and Knowledge, Attitude and Practice for Children Unintentional Injury (KAP) were also used as the protective factors. Risk factors index (RFI) and protective factors index (PFI) were computed in the study. Results: The severity of unintentional injury were positively correlated with SDQ, IBC and perceptions of risks and hazards(r=0.15, 0.23, 0.12, P<0.01), and were negatively correlated with HOME, PSAPQ and KAP(r=-0.25, -0.14, -0.09, P<0.01). Hierarchical linear regression showed that the total scores of SDQ, IBC and environmental of HOME predicted the severity of unintentional injury which could explain 34% variant of unintentional injury. It also indicated that the severity of unintentional injury were positively correlated with RFI (β=0.21) and negatively correlated with PFI(β=-0.18), the interaction was significant(β=-0.11,R2=0.31)(P<0.01). Conclusion Both risk and protective factors of unintentional injury have cumulative effects on the severity of injury among rural children. The relationship between risk factors and injury could be mediated by protective factors.

BACKGROUND: Pancreatic stellate cells (PSCs) activation plays a critical role in the development of chronic pancreatitis. Previous studies confirmed that thromboxane A2 receptor (TxA2r) was overexpressed in activated PSCs in rats. The purpose of this study was to investigate the role of TxA2r in the activation of PSCs induced by 8-epi-prostaglandin F2α (8-epi-PGF2α). METHODS: TxA2r expression in both quiescent and activated PSCs was detected by immunocytochemistry and immunoblot assay. Isolated PSCs were treated with 8-epi-PGF2α (10, 10, 10 mol/L) for 48 h, and SQ29548 (10, 10, and 10 mol/L), a TxA2r-specific antagonist for 48 h, respectively, to identify the drug concentration with the best biological effect and the least cytotoxicity. Then isolated PSCs were treated with SQ29548 (10 mol/L) for 2 h, followed by 10 mol/L 8-epi-PGF2α for 48 h. Real-time polymerase chain reaction was performed to detect the messenger RNA (mRNA) levels of α-smooth muscle actin (α-SMA) and collagen I. Comparisons between the groups were performed using Student's t test. RESULTS: TxA2r was up-regulated in activated PSCs in vitro compared with quiescent PSCs (all P < 0.001). Compared with the control group, different concentrations of 8-epi-PGF2α significantly increased mRNA levels of α-SMA (10 mol/L: 2.23 ± 0.18 vs. 1.00 ± 0.07, t = 10.70, P < 0.001; 10 mol/L: 2.91 ± 0.29 vs. 1.01 ± 0.08, t = 10.83, P < 0.001; 10 mol/L, 1.67 ± 0.07 vs. 1.00 ± 0.08, t = 11.40, P < 0.001) and collagen I (10 mol/L: 2.68 ± 0.09 vs. 1.00 ± 0.07, t = 24.94, P < 0.001; 10 mol/L: 2.12 ± 0.29 vs. 1.01 ± 0.12, t = 6.08, P < 0.001; 10 mol/L: 1.46 ± 0.15 vs. 1.00 ± 0.05, t = 4.93, P = 0.008). However, different concentrations of SQ29548 all significantly reduced the expression of collagen I (10 mol/L: 0.55 ± 0.07 vs. 1.00 ± 0.07, t = 10.47, P < 0.001; 10 mol/L: 0.56 ± 0.10 vs. 1.00 ± 0.07, t = 6.185, P < 0.001; 10 mol/L: 0.27 ± 0.04 vs. 1.00 ± 0.07, t = 15.41, P < 0.001) and α-SMA (10 mol/L: 0.06 ± 0.01 vs. 1.00 ± 0.11, t = 15.17, P < 0.001; 10 mol/L: 0.28 ± 0.03 vs. 1.00 ± 0.11, t = 11.29, P < 0.001; 10 mol/L: 0.14 ± 0.04 vs. 1.00 ± 0.11, t = 12.86, P < 0.001). After being treated with SQ29548 (10 mol/L) and then 8-epi-PGF2α (10 mol/L), the mRNA levels of α-SMA (0.20 ± 0.08 vs. 1.00 ± 0.00, t = 17.46, P < 0.001) and collagen I (0.69 ± 0.13 vs. 1.00 ± 0.00, t = 4.20, P = 0.014) in PSCs were significantly lower than those of the control group. CONCLUSIONS The results show that 8-epi-PGF2α promoted PSCs activation, while SQ29548 inhibited PSCs activation induced by 8-epi-PGF2α. The result indicated that TxA2r plays an important role during PSC activation and collagen synthesis induced by 8-epi-PGF2αin vitro. This receptor may provide a potential target for more effective antioxidant therapy for pancreatic fibrosis.

Objective To investigate the discrepancy of aldosterone synthesis process and potential regulation abnormality between aldosterone-producing adenoma(APA) and normal adrenal(NA) with microarray. Methods cRNA probes labelled with biotin were prepared from mRNA of APAs(APA group,n=10) or NAs(control group,n=7).The probes were hybridized with oligonucleotide microarray of target gene expression profile.Expression levels were read from the fluorescent intensity scanned.The difference of gene expression profile was analyzed by computer software.Differentially expressed genes were verified by real-time RT-PCR. Results Compared with control group,97 genes were up-regulated and 168 genes were down-regulated in APA group.In the genes related to steroid hormone synthesis,only CYP11B2 was significantly up-regulated.In the physiologic regulators of aldosterone synthesis,CYB5A,CYP17A1,DUSP1 and HMGCR were down-regulated,while RENBP and NR1H2 were up-regulated.As a key enzyme in the biosynthesis of cortisol,the expression of CYP17A1 gene was inhibited. Conclusion Among the aldosterone synthesis related enzyme and corresponding regulatory genes in APA,CYP11B2 may be a key synthetase,and the suppressed physiologic regulators of aldosterone synthesis may indicate the existence of neoplastic modulation.