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Article in Chinese | WPRIM | ID: wpr-849689


[Abstract] Objective To observe the effects of the adipocyte hormone leptin on GABA content and receptor expression in hypothalamus of mice with sleep deprivation, and explore the possible mechanisms. Methods Male C57BL/6 mice were randomly divided into three groups (8 each): control group, sleep deprivation (SD) group and leptin supplement (L-SD) group. Mice in control group were set up in a water environment without sleep deprivation, mice in SD group were set up in a "modified multi-platform water environment" to establish a sleep deprivation model, and mice in L-SD group were given leptin 1.3 mg/kg intraperitoneally twice daily in conjunction with sleep deprivation. Seven days after sleep deprivation, the general conditions of mice were observed, body weight was measured and hypothalamic tissues and plasma specimens were collected. ELISA was used to detect the plasma leptin levels, hypothalamic γ-aminobutyric acid (GABA) and glutamate (Glu) contents. Western blotting was performed to detect the expression levels of GABA key glutamate decarboxylase 67 (GAD67) and GABAA receptor α1 subtype protein (GABAARα1). Results Compared with control group, the weight of mice in SD group significantly reduced [(22.03±0.42) g vs. (17.75±0.75) g, P0.05). The hypothalamic Glu levels were obviously higher in SD group [(686.56±10.01) ng/g] and L-SD group [(668.64+9.93) ng/g] than that in control group [(577.11±16.36) ng/g] (P0.05). The expressive levels of GAD67 and GABAARα1 protein in the hypothalamus of mice in SD group [0.68±0.06, 0.69±0.07] were significantly lower than that in control group (1.09±0.13, 0.99±0.07) (P<0.05); While the expressive levels of GAD67 and GABAARα1 proteins in the hypothalamus of mice in L-SD group (1.39±0.19 and 1.33±0.14, respectively) were significantly higher than those in SD group and control group (P<0.05). Conclusion Leptin can up-regulate the expression of the key GABA synthase GAD67, increase the content of GABA and the expression of GABAARα1 protein in hypothalamus of sleep-deprived mice, which may be an important mechanism of leptin affecting sleep.

Article in Chinese | WPRIM | ID: wpr-817772


@#【Objective】To evaluate the effect of preoperative conjunctival sac irrigation with Anerdian Ⅲ disinfectant and its effect on the function of eye surface and tear film. 【Methods】 This clinical study was involved of thirty cases (30 eyes) undergoing phacoemulsification. Conjunctiva sac irrigation with Anerdian disinfectant (type III ,5g/L) was performed before the phacoemulsification procedure. Secretions of conjunctiva sac were examined by bacterial culture pre-and post-irrigation. Schirmer test,non-invasive first tear break-up time(NITBUT-F),non-invasive average tear break-up time(NITBUT-A),tear meniscus height(TMH),and goblet cell density(GCD)were also evaluated day 1 before and day 7 ,day 30 after operation.【Results】3 cases were positive in bacterial culture pre-irrigation ,while all were negative post-irrigation. There was no significantly change between Schirmer test and TMH from day 1 before ,day 7 and day 30 after operation . NITBUT-F,NITBUT-A,and GCD were significantly different between day 1 before and day 7 after operation(P<0.05),and shown no significantly difference between day 1 before and day 30 after operation.【Conclusions】 Our results suggested that conjunctiva sac irrigation with Anerdian disinfectant was efficient,while it might be related to the decrease of GCD and dysfunction of tear film. However,GCD would gradually improve as pre-operation and function of tear film could restore to normal.

Article in Chinese | WPRIM | ID: wpr-817740


@#【Objective】To detect the ability and the efficiency of recombinant PTD-HSP27 transport across RGC-5 cells and measure the role of recombinant PTD- HSP27 against oxidative stress damage induced by cobalt chloride on RGC- 5 cells,and explored its potential mechanism tentatively. 【Methods】 RGC- 5 cells were incubated with PTD- HSP27 labeled with FITC,followed by the observation of fluorescence using fluorescence microscope. We also lysed the cells by radio immunoprecipitation assay and measure the transport efficiency of PTD- HSP27- FITC through ultraviolet spectrophotometer subsequently. We established the RGC- 5 cell damage model. Cellular experiments were divided into three groups:normal control group,cobalt chloride damage group,PTD-HSP27 + cobalt chloride treatment group. Those three groups of cells were experimented using Annexin V- FITC/PI staining kit:to detect apoptotic cell ratio;Western Blotting:to detect the expression level of apoptosis related protein,including Bcl-2,Bax and Caspase-3.【Results】The fluorescence of PTD- HSP27- FITC was visualized inside the RGC- 5 cells using the fluorescence microscope,while the transport efficiency of PTD-HSP27-FITC was(47.29 ± 2.33)%. Cobalt chloride inhibited the survival vitality of RGC- 5 cells in a concentration- dependent manner. The comparison among normal control group,cobalt chloride damage group and PTD-HSP27+CoCl2 treatment group showed:PTD-HSP27 could effectively suppressed early apoptosis induced by cobalt chloride(P<0.01). Western Blotting results showed that PTD-HSP27 could effectively enhance the expression ratio of Bcl-2/Bax,and suppressed the activation of Caspase- 3(P<0.01).【Conclusions】Recombinant protein PTD- HSP27 could protect RGC- 5 cells against oxidative stress injury induced by cobalt chloride. Iit mainly regulated the expression ratio of Bcl-2/Bax,suppressed early apoptosis and improved cell viability in RGC-5 cells.