Objective:To explore the method of culturing rat hippocampal slice in vitro and to observe the changes of neurons' morphology and reactivity in cultured slice.Methods:Organotypic hippocampal slices were prepared by taking brains from 6-9 day old wistar rats.After the hippocampi was isolated and chopped into 400?m thickness,slices were transferred to Millipore inserts and placed in six-well trays in CO_2 incubators at 37℃.Then the cultured hippocampal slices were observed for the changes of morphology by macroscopic observation and using inverted phase contrast microscope;Expression of Fos protein in cultured hippocampal slice was detected by immunocytochemistry for observation of the internal response of hippocampal neurons to the damage;The electrophysiological activity of hippocampal neurons were detected by using patch clamp technique. Results:(1)The number of neurons in cultured hippocampal slices was increased,and the thickness of cultured hippocampal slices was decreased markedly along with the culturing period in vitro.After being cultured for 4 weeks,the thickness of hippocampal slices was decreased to about 150?m.(2)With the treatment of pilocarpine to induce seizure-like activity,the number of neurons which expressing Fos protein was increased in CA1 area of the cultured hippocampal slice.(3)With the application of pathch clamp technique,the changes of the ion currents of pyramidal neurons in CA1 area were recorded by the whole-cell recording after culturing for 1 week,2 weeks,3 weeks and 4 weeks respectively.Conclusion:Cultured hippocampal slice in vitro could retain satisfactory liveness and function for at least 4 weeks.

Objective:To explore the dynamic process of the expression of brain-derived neurotrophic factor(BDNF)in hippocampus cells after convulsion through in vitro and in vivo experiments. Methods:Seizures with duration of 30 minutes(30min SC) were induced in infant and adult Wistar rats(IRs&ARs) respectively. The rats were sacrificed at 6 different time points after SC termination.Dynamic change of BDNF level in happocampal cells was investigated by immunocytochemistry and ELISA. Primary hippocampal neurons cultured for 9 days were exposed to magnesium-free extracellular solution media for 3h to establish seizure-like discharge model. BDNF protein expressed in cultured hippocampal neurons was detected by immunocytochemistry. Results:(1)The cells expression of BDNF increased in CA1-CA4 and dentate of hippocampus after SC,and the most positive immunoreactivity was in dentate. The expression of BDNF was remarkably up-regulated at 3h(from 6.65?1.60 pg/?g to 11.22?2.44pg/?g) after SC in IRs group (P

Objective To investigate IL-6 and CRP levels,and their relationship with airflow obstruction、frequency of exacerbation in patients with COPD.Methods From Feb.2004 to Jul.2004,sputum and serum specimens were obtained from 54 patients with stable COPD and 10 age-matched healthy controls.The concentrations of sputum IL-6 and serum IL-6 were measured by ELISA.The concentrations of sputum CRP and serum CRP were measured by transmissive heterometric titration.Results (1)Sputum IL-6 levels in COPD patients of all stages were higher than those in controls(P

Iodine is an essential trace element for growth and development of fetus. More and more surveys currently show that the urinary iodine level of pregnant women is between 100 - 149 μg/L, which is lower than the level recommended by the World Health Organization (WHO). Under this level of iodine nutrition, different investigations have shown different effects on thyroid function of pregnant women, and the impact on growth and development of the fetus requires long-term objective evaluation. At present, there are three aspects: development, intelligence quotient (IQ), and psychological behavior. There are not many studies on comprehensive evaluation. This article combs the related published research to provide a reference for further clarifying the influence of iodine nutrition level during pregnancy on thyroid function of pregnant women and the growth and development of their offspring.

Objective To study the change of spleen Dendritic cells in normal rats treated by radio-frequency ablation(RFA). Methods Eighteen healthy SD rats were separated into group 1 week after RFA with 6 rats,group 2 week after RFA with 6 rats and control group with 6 rats. Spleen tissue were taken out respectively before RFA, 1 week after RFA and 2 weeks after RFA. The number and the phenotype of Dendritic cells in spleen were analyzed with flowcytometry. Results Pathologyical examination after RFA showed the characteristic that coagulation necrosis and cellular degeneration and granulation tissue forming appeared from target center to peripheral of the target. (10. 36±3. 21) % of normal rat mononuclear cells in spleen express OX-62, the ratio became (18. 03±5. 7) % 1 week after RFA and (12. 63±8. 0) % 2 weeks after RFA, the difference between group 1 week after RFA and control group was marked. (76. 33±7. 86) % of normal rat mononuclear cells in spleen express OX-6,the ratio became (78.33±7.25)% 1 week after RFA and (86. 04±7. 25) % 2 weeks after RFA, the difference between group 2 weeks after RFA and control group was marked. (63. 06±8. 77) % of normal rat mononuclear cells in spleen express CD86,the ratio was (55. 74±14. 49)% 1 week after RFA and (63.49±11.81)% 2 weeks after RFA,the difference between groups 1 week or 2 weeks after RFA and control group was not marked. Conclusions RFA can increasing the number of precursor Dendritic cells migrating from peripheral blood to spleen, and those cells may furtherly differentiate or maturate, which may be contributed to improve the ability delivery of body to antigen to a certain extent.

Objective To study changes in the expression levels of OX-62, OX-6 and CD86 of mononuclear cells and the related chemotatic factor macrophage inflammatory protein-1α (MIP-1α) in the livers of rats with Walker-256 tumors treated with radiofrequency ablation (RFA) and to elucidate the influence of RFA on differentiation and migration of liver dendritic cells(DCs).Methods Walker-256 liver tumors were induced in 60 SpragueDawley rats by implanting tumor particles. These rats were randomly divided equally into three groups from which liver tissue around the local area of the tumor was sampled at 7 d and 14 d after RFA. The mononuclear liver cells were separated with Ficoll density gradient centrifugation. The expression levels of OX-62, OX-6 and CD86 in the mononuclear cells were analyzed with flow cytometry. The expression level of MIP-1α in the liver tissue was detected by enzyme-linked immunosorbent assay (ELISA). Results The average expression of OX-6 in the control rats was 15.29 ±4.59% and those 7 d and 14 d after RFA were 34.20±11.62% and 39.18±9.14% respectively. The difference between the two RFA groups and the control group was statistically significant. The average expression rate of OX-62 in the control rats was 18.91±4.58% and those 7 d and 14 d after RFA were 24.49±4.45% and 22.77 ± 3.50% respectively. The difference between the 7 d group and the control group was significant. The average expression rate of CD86 in the control rats was 66.29±17.69% and those 7 d and 14 d after RFA were 55.29±10.69% and 55.93±12.64% respectively. These differences between both RFA groups and the controls group were not significant. The average expression level of MIP-1 α around the tumors was 232.92±54.58 pg/ml in the controls and 499.38±15.14 pg/ml and 495.90±9.94 pg/ml 7d and 14 d after RFA respectively. These differences from the controls were both statistically significant. Conclusion The expression of MIP-1α around the tumors was elevated after RFA, which could promote the migration of DCs. The changes in the expression of OX-62, OX-6 and CD86 also could reflect increased DC differentiation, which could improve local antigen-presenting capacity to a certain extent and recovery of host anti-tumor immune response.

Objective To investigate the modulating effects and explore their mechanism of 9-nitrocamptothecin and its liposomes to induce apoptosis and inhibit cell cycle in HepG2 and L02 cell lines. Methods Cells were incubated with 9-nitrocamptothecin(9NC) or with 9-nitrocamptothecin liposomes for 24 h, 48 h and 72 h, then, the cell viability was measured via MTT assay; cell cycle and apoptosis was evaluated by flow cytometry after stained by PI and Annexin V-PE/7AAD. Additional, Western Blot was used to evaluate the expression of cell cycle and apoptosis related protein. Results Both cells viability were apparently inhibited by the 9-nitrocamptothecin and 9-nitrocamptothecin liposomes, the inhibitory effect showed a time-dependent and dose-dependent manner. Both S and G2/M phases arrest were observed after incubated with drugs. HepG2 cell was completely arrested in S phase when 9NC concentration over than 0. 1 μmol/L after incubation for 24 h, while more than 95% cells arrested in G2/M phase when 9NC concentration is 0. 1 μmol/L after incubation for 72 h. Apoptosis induction effect also showed a time-dependent and dose-dependent manner. Western Blot results showed the expression of Bax and Caspase-3 were upregulated while Cyclin A, Cdk2, Cyclin E and Bcl-2 were downregulated. More importantly, the compounds were more cytotoxic to the cancer cell lines than to the normal liver cell. Conclusions 9-nitrocamptothecin and 9-nitrocamptothecin liposomes can potently inhibit cell growth via regulation of cell cycle and induction of apoptosis, and this effect was preferentially in cancer cell. Inhibitory of 9-nitrocamptothecin liposomes was slightly better than the 9-nitrocamptothecin.

Objective To observe the inhibitory effect and mechanism of 9-nitrocamptothecin liposomes on HepG2 liver carcinoma cells. Methods HepG2 cells were incubated with 9-nitrocampto-thecin(9NC) or with 9-nitrocamptothecin liposomes(9NC-LP) for 24 h, 48 h and 72 h. Cell viability was then measured by the MTT assay. Cell cycle and apoptosis were evaluated by flow cytometry.Western Blot was used to determine the expression of cell cycle and apoptosis related proteins. HepG2tumor-bearing mouse models were then established. The HepG2 tumor-bearing mice were randomly divided into control group, free liposomes group, DMSO group, 9NC low dose group, 9NC high dose group, 9NC-LP low dose group and 9NC-LP high dose group. There were 10 mice in each group.Drugs were administered by tail vein and tumor volume and body weight were observed 28 days after administration. Then animals were sacrificed and the expression of proteins from tumor homogenates was analyzed by Western blotting. Results In vitro, HepG2 cell viability was apparently inhibited by 9NC and 9NC-LP, and the inhibitory effect increased in a time-dependent and dose-dependent manner.Both S and G2/M phase arrests were observed after incubation with drugs. HepG2 cells were completely arrested in S phase with 9NC concentration over than 0.1 μmol/L after incubation for 24 h,while more than 95% of cells arrested in G2/M phase when 9NC concentration was 0.1 μmol/L after incubation for 72 h. In vivo, compared with the control group, the average tumor volume was reduced in both the 9NC and 9NC-LP group (P<0.05) , and the average animal body weight also decreased in both the 9NC and 9NC-LP group (P<0.05). There was no significant difference among the control group, free liposomes group, and DMSO group. The lights inhibition rates of tumor growth in the 9NC-LP(2.5 mg/kg),9NC-LP(1.5 mg/kg),and 9NC(1.5 mg/kg)groups were 87.02%, 51.57%and 35.47%, respectively. In the 9NC-LP(2.5 mg/kg)group, >50% of animals died 14 days after drug administration. Conclusion 9NC and 9NC-LP can inhibit HepG2 cell growth via cell cycle arrest and apoptosis induction. 9NC-LP has a more potent anti-tumor effect and fewer side effects in vivo,which means 9NC-LP is a promising compound for cancer therapy via intravenous administration.

Objective To investigate the incidence,correlative factors and prevention of perimenopausal symptoms.Methods 146 perimenopausal women were investigated by using Kupperman menopausal index(KMI),life event scale,improved social support scale and self-made inventory.Results We got 112 available data,the incidence of perimenopausal symptoms was 75.0%,58.0%was mild,15.2%was moderate,1.8%was sevcre.The main correlative factors of perimenopausal symptoms were inhabited environment,self emotional controllability,history of gynecologic diseases,history of depression,dysmenorrhea,premenstrual syndrome,menopause,score of life eventscale,improved social support scale.Conclusions The incidence of perimenopausal symptoms is high.It is also related to factors of society,family and mentality,except hypogonadism.Preventing perimenopausal symptoms according to its cirrelative factors may decrease its incidence and enhance the life quality of women.

Objective To determine the clinical characteristic, main treatment, and prognosis for the sake of more effective treatments for critically ill patients with H1N1 influenza. Methods Eight critically ill patients with H1N1 influenza in intensive care unit were retrospectively studied, including clinical characteristics, indexex of correlation, and prognosis. Results The acute physiology and chronic health evaluationⅡ score was 19.0±7.8. Five patients died, 4 of whom were caused by respiratory failure. The number of platelets in dead patient was lower than that in healing and improved ones(χ2=8.000,P＜0.05).All the 4 patients treated with glucocorticoid died, 5 out of the 6 patients received invasive mechanical ventilation rather than noninvasive mechanical ventilation, and 3 of them who complicated barotraumas in the lung died at last. Conclusion Critically ill patients with H1N1 influenza have high mortality. Respiratory failure is the main cause of death. Critically ill patients with H1N1 influenza should not be treated with glucocorticoid. Patients who need mechanical ventilation should be treated with invasive mechanical ventilation with low tidal volume and low positive end-expiratory pressure.