Evaluation Of Electro-acupuncture On Gas Signal Molecules In Cerebral Cortex Of Newborn Rat With HIE And Analysis Of Gene Mutations Of TSC And Phenotype Of Infantile Spasms In Children

Part one:Evaluation of electro-acupuncture on gas signal molecules in cerebral cortex of newborn rat with HIEObjective To investigate the change and mechanism.of carbon monoxide (CO), nitric oxide (NO) in cerebral cortex of newborn rat with hypoxic-ischemic encephalopathy by electrical acupuncture (EA) providing a theoretical basis of EA in treating cerebral palsy(CP).Methods The HIBD models were established and were randomly divided into eight groups (n= 12) (Sham, Sham-EA, HIBD, HIBD+EA, HIBD+SAM (S-adenosyl-L-methionine,), HIBD+SAM+EA, HIBD+HA (Hydroxylamine,), and HIBD+HA+EA). The motor function of rats was evaluated by suspension,slope experiment. CO content of cerebral cortex was determined by dual wavelength spectrophotometry,detection of NO content in cortex of nitrate reductase method. IHC staining was performed to observe expression of HIF-1α、 HO-1、CBS and NF-κB、nNOS in brain cortex and Western blot quantitative analysis as following. HE staining was used to observe the brain tissue morphology and morphological changes of nerve cells was analyzed by Nissl staining.Results (1) Results demonstrated that EA treatment promoted motor function of perinatal rats and the brain tissue morphology was improved. Similarly, EA treatment reduced CBS expression level, increase of expression level of HO-1 and HIF-la in cortex cells(P<0.05). The CO content of cortex cells in the EA treatment group significantly enhanced (**p< 0.01). (2)EA therapy significantly reduced the content of NO in cerebral cortex (*p< 0.05,**p< 0.01,) and paliated the nerve cells insult in HIBD rats, decreasing expression level of nNOS and NF-κB in brain cortex cells (*p< 0.05,**p< 0.01). (3)The inhibitor(HA) and agonist(SAM) of CBS respectively alleviated and worsened the brain injury.Conclusion (1)EA therapy might be ascend CO content of cortex to relieve hypoxic brain injury by the hydrogen sulfide/CBS-CO/HO-land HIF-la pathway. (2) EA treatment reduced cortical NO generated to ease brain damage via NF-KB/nNOS system.Part two:Analysis of gene mutations of TSC and phenotype of Infantile Spasms in childrenObjective This study investigates the novel mutation in one of two TSC genes in tuberous sclerosis children with or without infantile spasms (IS), and further studying of their correlation. Methods 48cases, who were diagnosed TSC, were divided into TSC in relation with IS group and without IS group. DNA sequencing analyzing of TSC gene mutation, phenotype, electroencephalogram,brain MRI or CT, ultrasonic of organs,etc,Which were analysed, were performed in 48cases. Results In 22 cases TSC children with IS,18 exists TSC2 gene mutation (including 11 novel mutations:) and 3 exists TSC1 gene mutation (including 2 novel mutations). Similarly,21 exists TSC2 gene mutation (including 11 novel mutations) and 5 exists TSC1 gene mutation (including 4 novel mutations) in 26 cases TSC children without IS. The TSC genes mutation is associatesd with TSC and several novel mutations are found. Conclusion However, in all new discovered mutations, no difference in TSC children with or without IS. It is essential that further increasing quantity samples were analyzed to provided some new targets for the treatment of TSC with or without IS.