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Objective To explore the damage of spiral ganglion neurons(SGNs),the protective effects of different dosages of healthy ear compound (HEC)from traditional Chinese medicine(TCM) against age-induced SGNs degeneration and its possible mechanism in spiral ganglion neurons(SGNs)of C57BL/6J mice.Methods Totally 36 C57BL/6J mice just after ablactation were randomly divided into four groups.Normal control group (n =6)drank tap water daily from ablactation to 2 months old.Ageing-related SGNs apoptosis control group(n=12)drank tap water daily from ablactation to 7 months old.High dose TCM group(n=12)at drank 3.65 g/kg/d of HEC from ablactation to 7 months old.Low dose TCM group(n=6)drank 0.91 g/kg/d of HEC from ablactation to 7 months old.The animal cochleae were immediately removed at the termination of the experiment.In each group,the cochleae of 6 animals were used for paraffin embedding,slicing and toluidine blue staining to observe neuronal morphological changes.The caspase-3 mRNA expression study was performed by real-time PCR technique in 6 cochleae of High dose TCM group and ageing-related SGNs apoptosis control group.Results The morphological structure of cochlear SGNs represented healthy and normal density in normal control group at 2 months old.In contrast,amount or density of SGNs in cochlear basilar part was significantly damaged and reduced in ageing-related SGNs apoptosis control group at 7 months old(P＜ 0.001).But the high dose TCM group at 7 months of age was similar to the normal control group at 2 months old in morphological structure,amount or density of SGNs(P＞0.05).The low dose TCM group was significantly different from other 3 groups in amount or density of SGNs (P＜0.001).However,SGNs in the middle part and apical part showed integrity in each group.In addition,the expression level of caspase-3 in the cochlea of high dose TCM group was also obviously different with age-related SGNs apoptosis control group(P＜0.01) Conclusions Ageing-related damage of SGNs in C57BL/6J mice begins from the base of cochlea and progresses towards the apex.The HEC of TCM could significantly protect SGNs against age-induced apoptosis in SGNs.The efficacy of the high dose TCM is better than that of the low dose TCM.Its SGNs protective mechanisms might be related to involving the caspase-mediated cell apoptotic pathway.
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OBJECTIVE To study the correlation of hepatitis B virus(HBV)basic core promoter(BCP) mutation with the interleukins(IL-10,IL-12),tumor necrosis factor(TNF)-?,interferon(IFN)-?,as well as HBV DNA contents in patients with chronic hepatitis B virus infection.METHODS(1) Project subject: 176 patients(chronic hepatitis B with mild,moderate and severe degree;liver cirrhosis,chronic fulminant and hepatocellular carcinoma(HCC)) with chronic hepatitis B virus infection were studied.(2) Project methods: ① The A to T mutation at nucleotide 1762 and G to A mutation at nucleotide 1764 were determined by the method of polymerase chain reaction(PCR) microplate hybridization ELISA in these patients.② The serum cytokines(IL-10,IL-12,TNF-? and IFN-?) of these patients were measured by specific-ELISA.RESULTS The serum levels of cytokines(IL-10(80.96?30.86 vs 72.11?24.19 mg/L),IL-12(41.33?15.10 vs 35.98?14.47 mg/L),TNF-?(56.04?27.05 vs 38.01?10.49 mg/L),IFN-?(19.81?12.29 vs 16.55?8.99 mg/L))and HBV DNA contents(108.2478?0.9826 vs 105.8876?1.4822copies/ml) in BCP mutant group were significantly higher than that in non-mutant group(P
ABSTRACT
Objective: To observe the preventive effect of Bu Shen Jian Pi Capsule on age-related cochlear hair cells apoptosis in C57 BL /6J mice,and research its mechanism of action.Methods: 30 new born C57 BL /6J mice were divided into 2 groups,with 10 animals in control group(CG) and 20 animals in Bu Shen Jian Pi Capsule (experimental group,EG).The animals in CG were fed regularly,while those in EG were fed regularly at the first one months after the birth and then,commonly fed water was substituted by the medical solution of Bu Shen Jian Pi Capsule for these animals in EG.The experimental course lasted for seven months.The cochleas of the two groups of mice were taken out for cochlear surface preparation,then being observed under optical microscope with a comparative analysis on the status of degenerative changes in the hair cells of cochlear between the tow groups.Results: Cochlear hair cells damage of CG animals showed aggravating tendency with the increase of the age,and followed the developmental rules of lesion progression in cochlear hair cells from the basal turn to the apical turn and with the number of outer hair cells loss greater than that of inner hair cells.In contrast,the cochlear damage of animals of EG was relatively milder than that in animals of CG,compared among the animals at the same age between them.Conclusion: This study showed that Bu Shen Jian Pi Capsule could postpone age-related cochlear hair cells apoptosis in C57 BL /6J mice,therefore,it suggested that Bu Shen Jian Pi Capsule might possess the pharmacological action of preventing presbyacusis.We assume the possible protective mechanism of Bu Shen Jian Pi Capsule might be related to increase of cells cAMP,regulation of metabolism of nucleic acid,and protectoin or stabilization of DNA structure.
ABSTRACT
Objective: A traditional Chinese medicine(TCM) transformed from Jian Er Ⅱ,Cong Er capsule was studied in protection against age-induced degeneration in cochlear spiral ganglion neurons and auditory nerve fibers in C57 BL /6J mice.A possible protective mechanism was also discussed.Methods : 40 neonatal C57 BL /6J mice were randomly divided into 2 groups with an average of 20 animals in each group.The animals in control group were fed with regular mouse feedstuff and regular water after ablactation,while animals in experimental group were fed the same mouse food after ablactation,but drink the medical solution of Cong Er capsule instead of regular water from two months of age.Animals were terminated at 7 months of age.The cochlea were removed and processed for a series of modiolus sections.The density of spiral ganglion neurons in Rosenthal's canal in different turns was counted,and the auditory nerve fibers in habenula perforata at basal turn and apical turn were also quantified respectively.Statistical comparative analysis was performed between the two groups.Results: Density of spiral ganglion neurons in cochlear basal turn in control group was significantly reduced.In survival ganglion neurons,an obvious degeneration was observed in control group.The number of auditory nerve fibers in cochlear basal turn also decreased.In contrast,Cong Er capsule treated animals had an increase in survival ganglions and auditory nerve fibers,especially,a significant difference was seen in density of spiral ganglion neurons in the basal turn between the two groups(P