1.Study on the expression of BDNF after status convulsion in wistar rats and seizure-like discharge of cultured hippocampal neurons
Journal of Chongqing Medical University 1986;0(03):-
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
2.A study for morphology and the reactivity of rats hippocampal slice in vitro culture
Journal of Chongqing Medical University 2003;0(05):-
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.
3.Levels of interleukin-6 and CRP in patients with chronic obstructive pulmonary disease and its significance
Chinese Journal of Practical Internal Medicine 2006;0(13):-
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
4.The effect of pregnant women's urinary iodine at 100 - 149 μg/L on thyroid function and offspring
Chinese Journal of Endemiology 2021;40(5):426-430
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.
5.Changes of macrophage inflammatory protein-1α in the livers of rats with Walker-256 tumors treated with radiofrequency ablation
Weide DAI ; Zhihui FAN ; Minhua CHEN ; Fujin HE ; Hongmin LI
Chinese Journal of Physical Medicine and Rehabilitation 2010;32(8):574-577
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.
6.Effect of silencing of HIF-2α gene on chemosensitivity of human breast carcinoma cells
Na LI ; Guoyang HE ; Yongzhen LI ; Zhihui WANG
Chinese Journal of Clinical and Experimental Pathology 2017;33(5):501-504
Purpose To investigate the effects of hypoxia inducible factor-2α (HIF-2ot) siRNA on proliferation and chemotherapy sensitivity of breast carcinoma MCF-7.Methods RNA interference was used to silence the expression of HIF-2α in MCF-7 cells.The changes of HIF-2α gene expression were detected by immunocytochemistry and RT-PCR.Under hypoxia environment simulated by CoCl2,MTT assay and flow cytometry (FCM) were used to measure cell growth inhibition rate and cell apoptosis of MCF-7 cells under different dosages of chemotherapeutic agents (5-fluorouracil,adriamycin,and paclitaxel).Results Expression of HIF-2α in MCF-7 were down-regulated by HIF-2α siRNA (P < 0.05).The proliferation inhibition and apoptosis rates were evidently increased after transfection with HIF-2α siRNA (P < 0.05),chemotherapy drug sensitivity was enhanced.Conclusion HIF-2α siRNA can induce the apoptosis and inhibit the proliferation and enhance the sensitivity of breast carcinoma MCF-7 cell line to chemotherapeutic agents.Blocking HIF-2α maybe a very promising strategy for breast carcinoma gene therapy in combination with chemotherapy.
7.Modulating effects of 9-nitrocamptothecin and its liposomes to induce apoptosis and inhibit cell cycle in HepG2 and L02 cell lines
Shunzhen ZHENG ; Jinli LU ; Zhihui CHEN ; Bin HE ; Shengquan ZOU
Chinese Journal of Hepatobiliary Surgery 2011;17(5):416-419
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.
8.In vitro and in vivo inhibitory effects and mechanism of 9-nitrocamptothecin liposomes on HepG2 cell
Shunzhen ZHENG ; Jinli LU ; Zhihui CHEN ; Bin HE ; Shengquan ZOU
Chinese Journal of Hepatobiliary Surgery 2011;17(7):571-575
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.
9.Bond strengths of absorbable polylactic acid root canal post with three different adhe-sives
Hui PAN ; Can CHENG ; Jia HU ; He LIU ; Zhihui SUN
Journal of Peking University(Health Sciences) 2015;47(6):990-993
Objective:To find absorbable adhesives with suitable bonding properties for the absorbable polylactic acid root canal post. To test and compare the bond strengths of absorbable polylactic acid root canal post with three different adhesives. Methods:The absorbable polylactic acid root canal posts were used to restore the extracted teeth, using 3 different adhesives: cyanoacrylates, fibrin sealant and glass ionomer cement. The teeth were prepared into slices for micro-push-out test. The bond strength was statistically analyzed using ANOVA. The specimens were examined using microscope and the failure mode was divided into four categories:cohesive failure between absorbable polylactic acid root canal posts and adhesives, cohesive failure between dentin and adhesives, failure within the adhesives and failure within the absorbable polylactic acid root canal posts. Results:The bond strength of cyanoacrylates [(16. 83 ± 6. 97) MPa] and glass ionomer cement [(12. 10 ± 5. 09) MPa] were significantly higher than fibrin sealant [ ( 1 . 17 ± 0 . 50 ) MPa ] , P <0 . 001 . There was no significant difference between cyanoacrylates and glass ionomer cement (P =0. 156). In the group of cyanoacrylates, the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives was 25 . 0%, the cohe-sive failure between the dentin and the adhesives was 16. 7%, the failure within the adhesives was 33. 3%, and the failure within the absorbable polylactic acid root canal posts was 25 . 0%. In the group of fibrin sealant , the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives was 66 . 7%, the cohesive failure between the dentin and the adhesives was 22 . 2%, the failure within the ad-hesives was 11. 1%. In the group of glass ionomer cement, the cohesive failure between the absorbablepolylactic acid root canal posts and the adhesives was 87. 5%, the failure within the adhesives was 12. 5%. The major failure mode in fibrin sealant and glass ionomer cement was the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives. No major failure modes were found in the group of cyanoacrylates. Conclusion:The bond strength of fibrin sealant is low, which cannot meet the requirement of clinical use. The bond strengths of cyanoacrylates and glass ionomer cement are suitable for clinical use. The cyanoacrylates are a kind of absorbable adhesive which has suitable bonding proper-ties for the absorbable polylactic acid root canal post.
10.Pharmacodynamics interaction between propofol and remifentanil under different harmful stimulations analyzed by response surface methodology
Zhihui LIAO ; Nong HE ; Yonghua YU ; Ming YI
The Journal of Clinical Anesthesiology 2016;32(4):354-357
Objective To observe the patients hemodynamic reactions to the different stimula-tions under different blood concentrations of propofol with remifentanil.Exerting the response surface methodology,the half of the maximum effective concentration of propofol and the interaction index of propofol with remifentanil were calculated.Methods A total of 120 patients for scheduled abdominal surgery,randomly divided into 12 groups(n = 10).TCI 12 different concentration combinations of propofol with remifentanil,also the the hemodynamic reactions to harmful stimulations of intubation and incising were observed.The data were analyzed by responding surface analysis.Results The in-teraction index of propofol and remifentanil under intubation was 0.523 0±0.223 7,that under inci-sing 0.361 3±0.148 4,half of the maximum effective concentration of propofol for intutation and in-cising was (6.878 7 ± 1.832 1 ) μg/ml and (4.41 1 6 ± 0.902 5 ) μg/ml respectively. Conclusion Propofol and remifentanil have a synergistic inhibitory effect on harmful cardiovascular reaction among Chinese.During the endotracheal intubation,the requirements of half of the maximum effective concentration of propofol and intensity of the synergy with remifentanil are greater than those during the incising.