ABSTRACT
Objective To identify the host-cell death pathways (apoptosis, autophagy or necrosis) in L929 cells at the time point of 48 hours post infection (h.p.i.) with Chlamydia muridarum.Methods L929 cells were infected with Chlamydia muridarum at a multiplicity of infection (MOI) of 0.85 for 48 hours.Nuclear fragmentation was observed under fluorescence microscopy following staining L929 cells with DAPI (4′,6-diamidino-2-phenylindole).L929 cells were stained with propidium iodide (PI) plus Annexin Ⅴ and then analyzed by fluorescence-activated cell sorting (FACS) to clarify whether apoptosis or necrosis occurred after Chlamydia muridarum infection.L929 cells were transiently transfected with GFP-LC3 and observed under fluorescent microscopy to analyze cell autophagy.Western blot assay was performed to detect LC3 protein for further analysis of autophagy.Results Apoptosis was not induced in L929 cells by Chlamydia muridarum infection at 48 h.p.i.as no significant nuclear fragmentation was observed.Results of FACS showed that most cells died due to necrosis.Moreover, fluorescent dots of GFP-LC3 formed after infecting transfected L929 cells with Chlamydia muridarum.An increased ratio of LC3Ⅰ to LC3Ⅱ in the L929 cells infected with Chlamydia muridarum was detected by Western blot assay, indicating that autophagy occurred during Chlamydia muridarum infection.Conclusion Necrosis and autophagy rather than apoptosis are induced in L929 cells 48 hours after infection with Chlamydia muridarum.
ABSTRACT
Objective To evaluate the clinical efficacy of the intratympanic inflation by tympanum inflatable balloon for patients with Meniere’s disease(MD) .Methods A total of 36 confirmed cases (according to the listening stage) of Ⅱ - Ⅲ period of MD patients were randomly divided into two groups :experimental group and control group ,18 cases of matched group ,who were staged before preoperation by hearing ,Ⅱ period (average threshold 25~40 dB HL) 6 cases ,Ⅲ period (41~70 dB HL) 12 cases ,were treated by intratympanic methylprednisolone per‐fusion after putting tympanostomy tube under endotoscope ,then once every 3 days perfusion with a total of three times .There were 18 cases of experimental group staged before preoperation by hearing ,Ⅱ period (average thresh‐old 25~40 dB HL) in 5 cases ,Ⅲ period (41~70 dB HL) 13 cases treated by using tympanum inflatable balloon in the tympanum under endotoscope .Two groups were given conventional improved microcirculation and nutritional therapy .Results In 18 cases of patients in experimental group ,for the 18 to 24 months follow -up after treatment ,6 cases of vertigo were fully controlled (grade A) ,10 cases of vertigo basically controlled (grade B) ,2 cases of vertigo partly controlled (grade C) .Vertigo effective control rate was 88 .89% (16/18) .After 18 to 24 months treatment ,two groups of 36 patients with tinnitus and pure tone average threshold had no obvious change .In 18 ca‐ses of patients in the matched group ,during the 18 to 24 months follow -up after treatment ,3 cases of vertigo were fully controlled (grade A) ,7 cases of vertigo basically controlled (grade B) ,8 cases of vertigo partly controlled (grade C) .Vertigo effective control rate was 55 .56% (10/18) .The experimental vertigo basic control rates were higher than the control group (χ2 = 4 .985 ,P<0 .05) .The activitives in the two groups had improved significantly . No obvious differences were found between two groups .Conclusion With the intratympanic gasing by tympanum in‐flatable balloon for patients of Ⅱ - Ⅲ Menier’s disease ,control rates was high and motile ability were improved , but tinnitus and hearing improvement was not obvious .