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OBJECTIVE： To study on the effects of prophylactic administration of Ramulus mori polysaccharides （RMP） on inflammatory response of renal ischemia reperfusion injury （RIRI） model mice and to explore its possible mechanism. METHODS： Totally 60 C57BL/6 mice were randomly divided to sham operation group， model group， atorvastatin group （positive control， 15 mg/kg）， RMP low-dose， medium-dose and high-dose groups （300， 600， 1 200 mg/kg）. Except for sham operation group， RIRI model was induced in other 5 groups. 24 h before surgery， they were given relevant medicine intragastrically， once a day， for consecutive one week. 24 h after reperfusion， the mice were sacrificed. The serum levels of Scr and BUN were detected. The morphological changes of renal tissue were observed under optical microscope. The serum levels of IL-1β， IL-6， IL-10 and TNF-α were determined by ELISA. Western blot assay was used to determine the protein expressions of Toll-like receptor 4 （TLR4）， p38 mitogen-activation protein kinase （p38MAPK） and p-p38MAPK. RESULTS： Compared with sham operation group， the serum levels of Scr and BUN were significantly elevated in model group （P＜0.01）. RIRI led to typical inflammatory response of renal tissue， widespread renal tubular epithelial cell degeneration and necrosis， and inflammatory cells infiltration. Serum levels of IL-1β， IL-6， IL-10 and TNF-α were increased significantly （P＜0.01）. The protein expressions of TLR4， p38MAPK and p-p38MAPK were increased significantly in renal cortex （P＜0.01）. Compared with model group， serum levels of Scr and BUN were decreased significantly in administration groups （P＜0.05 or P＜0.01）. The pathological damage of renal tissue was improved in varying degrees， especially in the RMP medium-dose and high-dose groups. Serum levels of IL-1β and IL-6 were decreased significantly in administration groups （P＜0.05 or P＜0.01）. Serum levels of IL-10 were further increased in atorvastatin group and RMP high-dose group （P＜0.01）， and serum level of TNF-α was decreased significantly in atorvastatin group and RMP medium-dose and high-dose groups （P＜0.05 or P＜0.01）. The protein expressions of TLR4 and p-p38MAPK in renal cortex were decreased significantly in administration groups （P＜0.05 or P＜0.01）. CONCLUSIONS： RMP prophylactic administration can improve RIRI of mice， the mechanism of which may be associated with relieving the inflammatory response through inhibition of TLR4/p38MAPK signaling pathway.
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OBJECTIVE:To study the protective effect of fingolimod on renal ischemia reperfusion injury (RIRI) model mice and its mechanism.METHODS:A total of 60 mice were randomly divided into sham operation group,model group,fingolimod group (1 mg/kg) and fingolimod+wortmannin group [fingolimod 1 mg/kg+phosphatidylinositol 3-kinase (PI3K) specific blocker wortmarmin 1.4 mg/kg],with 15 mice in each group.Except for sham operation group,RIRI model was induced in other 3 groups,and those model mice were given relevant medicine via caudal vein at once 24 h before surgery.Serum of mice were collected in each group after 24 h perfusion.Serum levels of Scr and BUN were measured by automatic biochemical analyzer.The pathological changes of renal tissue were observed under light microscope.The protein expression of intercellular cell adhesion molecule-1 (ICAM-1),monocyte chemoattractant protein-1 (MCP-1) and phosphorylated protein kinase B (p-Akt) in renal tissue were measured by Western blot assay.RESULTS:Compared with sham operation group,the serum levels of Scr and BUN in model group were increased significantly (P＜0.01).Pathological changes were found in the kidney,and RIRI led to widespread renal tubular epithelial cell injury,apoptosis and inflammatory cells infiltration.The protein expression of ICAM-1 and MCP-1 in renal tissue were increased significantly (P＜0.01),the protein expression of p-Akt was increased slightly (P＞0.05).Compared with model group,other indexes of fingolimod group were improved significantly (P＜0.01) except that the protein expression of p-Akt in renal tissue was increased significantly (P＜0.01).Compared with fingolimod group,above indexes of fingolimod+wortmannin group were reversed (P＜0.05 or P＜0.01).CONCLUSIONS:Fingolimod can obviously ameliorate renal injury induced by RIRI in mice,the mechanism of which may be associated with the activation of PI3K/Akt signaling pathway.
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Objective To investigate the patterns of gait and locomotion in three dimension space using OptiTrack motion capture system in 30 healthy people aged 20-70 years. Methods The 3D OptiTrack motion capture system was applied in young(aged 20-30 years,n=10),middle-aged(aged 40-50 years,n=10)and the elderly (aged 60-70 years,n=10)to capture the gait and the rhythmic oscillations of the trunk,head,hip and kneewhen subjects walked on the treadmill under three different conditions of normal walking,walking under visual interfering and walking under proprioceptive interfering.Several markers were placed on the subject.For each locomotor trial, the preferred walking speed,stride width,stride time,the absolute angular dispersions and the stander deviation of four segments around the roll,pitch and yaw axes,and gait instability(λ)were calculated to assess the equilibrium strategies of head,trunk,pelvis and knee under different experimental conditions. Results With increasing age, there were decreases in preferring walking speed(PWS)and increases in stride width(F=3.23,3.87;P=0.030, 0.020,respectively).The absolute angle of the segment of head in roll axis was significantly smaller in young group than in middle age and elderly group(F=4.01,3.72;P=0.035,0.028,respectively)under the condition of normal walking and proprioceptive interfering.The trunk sway in yaw was significantly decreased while significantly increased in roll plane either in normal walking or walking under proprioception interfering(F=3.19,P=0.015) .The standard deviation(SD)of movement of head,trunk and hip in yaw direction was higher in young group than in middle-age and elderly group(F=2.89,3.14,3.92;P=0.036,0.019,0.034,respectively),but was lower in young group in roll direction than in middle-age and elderly group(F= 3.90,2.56,3.28;P= 0.017,0.021,0.056, respectively),the difference existed only in hip and trunk under proprioception interfering.The SD value of movement of bilateral knee joints was increased in elderly subjects.Using mediolateral-λ(ML-λ)to predict the gait stability,the results showed a significantly increased λ value by our small circle,and the λ value was positively correlated with age(r=0.03,P<0.05). Conclusions With aging,the abnormal adjustment of body posture and the incongruity of gait may cause a high risk for falling,and gait instability begins to increase as early as age 40-50 years.This finding supports that local dynamic stability is likely to be an indicator of falling risk.OptiTrack motion capture system can be used to evaluate a balance for rehabilitation.
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OBJECTIVE To explore the expression of Bcl-2, on mRNA and protein levels, in the different age of C57BL/6 mice cochleae and the expression localization in the cochleae.METHODS Using ABR to test the hearing level in C57BL/6 mice. Surface preparation of cochlear basil membrane is used to observe the morphology and amout of the outer and inner hair cells in different age of C57BL/6 mice. Fluorescent quantitative real time PCR, immunofluorescence histochemical method and western blot are used to detect the expression of Bcl-2 on the mRNA and protein levels in the C57BL/6 mice cochlea of different age groups ('young group', 'elderly group').RESULTS ABR results showed that the hearing threshold of 'older' C57BL/6 mice is much higher than that in the 'young' mice, and surface preparation of cochlear basil membrane showed the hair cell localized in the cochlear basil turn of 'old' mice arranged in a disorder station and part of hair cells were lost. Also, the spiral ganglion cells arranged sparsely and messily. Fluorescence quantitative real-time PCR results suggest the expression of Bcl-2 on/at the mRNA level of the 'old' mice cochleae decreases significantly, compared to that in the 'young' mice. The results of Immunofluorescence and Western blot suggest the expression of Bcl-2 on/at the protein levels of the 'old' mice cochlea decreased, compared to that in the 'young' mice. Also, the Bcl-2 is located in the cytoplasm, and the expression of Bcl-2 in the inner hair cells seems higher than that in the outer hair cells. CONCLUSION The expression of Bcl-2 significantly deceased in the 'old' C57BL/6 mice cochleae, both on mRNA and protein level, which may be related to the hearing loss and loss of hair cells.
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OBJECTIVE To explore the role of miR-34a on upregulating the expression of Bcl-2,which induces the apoptosis of primary auditory cortex neuron in the central mechanism of age-related hearing loss.METHODS Using C57BL/6 suckling mouse to obtain the brain tissue from auditory cortex according to the Location map,and to primary culture the neurons.After transfection on primary neurons,western blot and immunofluorescence were used to detect the expression of Bcl-2 and then hoechst staining was used to detect the apoptosis after transfection.RESULTS primary culture of auditory cortex neurons abtained from enzyme digestion were transfected with miR-34a mimic and miR-34a inhibitor to upregualte or downregulate the expression of miR-34a,the results showed that the expression of Bcl-2 was decreased after upregulation of miR-34a with the concentration of 5 nmol/L(t=5.127,P＜0.05),there was significant difference between the concentration of 10 and 20 nmol/L(t=6.379,P＜0.05),while increased after downregulation of miR-34a with concentration of 5 nmol/L(t=4.926,P＜0.05),there was significant difference between the concentration of 10 and 20 nmol/L(t=5.821,P＜0.05).Hoechst staining showed that the apoptotic neurons was increased after transfection of miR-34a mimic.CONCLUSION miR-34a induce the auditory cortex neuron apoptosis through downregulates Bcl-2,which central mechanism of age-related hearing loss.
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Objective To study the relationship between tubomanometry(TMM) and tympanograms before and after the treatment of otitis media with effusion(OME) in children,and to evaluate the function of tubomanometry in forecasting the prognosis of OME.Methods Data from 66 patients(98 ears) were collected before and after one to four weeks treatment,all the patients received tympanometry and tubomanometry ETS,recorded eustachian tube scores(ETS) and tympanogram types.Results Before treatment,there were significant differences between the ETS of normal ears and sick ears,also between ETS of type B tympanograms and type C.There was significant difference between ETS before and after treatment,but not between the ETS of type B tympanograms and type C after treatment.The increased ETS value of type B tympanograms was higher than type C with a significant difference.There was a significant difference between the ETS of the effective group and the invalid group before and after treatment,whether type B tympanograms or type C.Conclusion Eustachian tube scores increased after treatment,and the functions of eustachian tube were improved in children of otitis media with effusion.Before treatment,the ETS of type B tympanograrns was lower than type C,indicating a poorer function of eustachian tube.After treatment,the increased ETS value of type B tympanograms was more than type C.Tubomanometry could forecast the prognosis of otitis media with effusion in children.
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Objective:To investigate the status of the vestibular function of the patients with chronic positional symptoms after peripheral acute vestibular syndrome (AVS) and the curative effect of the vestibular rehabilitation therapy (VRT). Method:Using caloric test (CT), head shaking nystagmus test (HST), cervical vestibular evoked myogenic potentials as well as ocular vestibular evoked myogenic potentials to estimate the function of semicircular canal and otolith organs. The patients with normal VEMPs are divided as Group A. Otherwise are as Group B. Both groups are treated with VRT. The curative effect is estimated by vestibular symptom index (VSI) and Berg balance scale (BBS). Result:Thirty-three of 37 patients (86.5%) had an abnormal result of CT and HST, with 23 of these patients (65.7%) had an abnormal of both test. Twenty-two patients (59.5%) were in Group A and 15 (40.5%) in Group B. Before the therapy, Group B had a higher score of the balance and dizziness symptoms of VSI (P<0.05), and Group A had a higher score of the BBS (P<0.05). After the therapy, the VSI scores of both groups dropped and scores of the BBS raised. Conclusion:Patients with chronic positional symptoms after peripheral AVS have dynamic vestibular lesions to different extents. Those with otolith organs lesions tend to have a worse function of balance. Nevertheless, patients have a better off after VRT.
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OBJECTIVE@#To investigate the risk factor,type and characteristic nystagmus of the otolith abnormal migration during diagnosis and treatment for posterior semicircular canal benign paroxysmal positional vertigo (PSC-BPPV). The therapy and prevention is also discussed.@*METHOD@#Four hundred and seventy-nine patients with PSC-BPPV were treated by Epley's canalith repositioning procedures(CRP) from March 2009 to March 2012. We observed otolith abnormal migration complicating during diagnosis and treatment. According the type of otolith abnormal migration, the additional repositioning maneuver was performed.@*RESULT@#The rate of complication was 8. 1%(39/479), with canal conversion in 5.4%(26/479) and primarily canal reentry in 2.7%(13/479). The rate of incidence of conversion to horizontal canal conversion and anterior canal were 4. 8%(23/479)and 0. 6%(3/479) respectively. All the patient was cured in follow up. The risk factors were unappropriated head movement during or after CRP, including another Dix-Hallpike were performed immediately.@*CONCLUSION@#To prevent the complications,the pathognostic positioning sequence and angle of head rotation are commenced during CRP. Appropriate short time postural restrictions post-treatment is necessary. Careful observation of nystagrnus variation is crucial to determine the otolith abnormal migration.
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Humans , Benign Paroxysmal Positional Vertigo , Therapeutics , Head , Incidence , Nystagmus, Pathologic , Otolithic Membrane , Patient Positioning , Semicircular Canals , VertigoABSTRACT
Objective To explore the possible mechanism of miR 34a/Bcl-2 apoptotic pathway in the mouse model of age-related hearing loss.Methods A C57BL/6 mouse model of age-related hearing loss was conducted,and 4-week-old and 12-month-old mice were considered as the objects.The auditory brainstem response (ABR) was used to test the hearing function.The TdT-mediated dUTP nick end labeling (TUNEL) was used to observe the apoptosis of neuron in auditory cortex.The mRNA and protein levels of miR-34a,Bcl-2 and caspase 3 were detected by real-time PCR and Western bloting,respectively.Results The ABR showed that the hearing threshold level at 4,8,16,32 kHz were higher in 12-month-old mice than in 4-week-old mice [(80.0±2.5) dBHL vs.(32.0 ±4.5) dBHL,(74.0±3.5) dBHL vs.(51.0±1.2) dBHL,(86.0±4.6) dBHL vs.(51.0±3.5) dBHL,(87.0±6.6) dBHL vs.(56.0±1.5) dBHL,all P＜0.05].Compared with 4 week-old mice,the total number of neurons in auditory cortex was decreased,the number of apoptosis neurons was increased,the expressions of miR-34a (t=6.02,P=0.001),Bax (t=6.51,P=0.012) and Caspase 3 (P=0.023) rised,and the expression of Bcl-2 (t=7.12,P=0.032) declined in 12 month-old mice.Conclusions The activation of miR-34a/Bcl-2 apoptotic pathway may be one of the mechanisms of age-related hearing loss.
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Aim To evaluate the effects of notoginsen-oside R1 on store-operated calcium entry ( SOCE ) in pulmonary arterial smooth muscle cells ( PASMCs ) of chronic hypoxia ( CH)-and monocrotaline ( MCT)-in-duced pulmonary hypertension ( PH) rats. Methods Mn2+ quenching of Fura-2 and measurement of intra-cellular free calcium concentration ( [ Ca2+] i ) using fluo-3 were examined in PASMCs of CH-exposed and MCT-treated rats. Results ①CH-exposed and MCT-treated rats exhibited profound PH when examined 3 weeks after hypoxia exposure or MCT injection, respec-tively. ②In the presence of 3 μmol·L-1 nifedipine, 10 μmol · L-1 notoginsenoside R1 significantly re-duced cyclopiazonic acid ( CPA )-induced the percent reduction in Fura-2 fluorescence measured 500 sec af-ter application of Mn2+, the maximal rate of Mn2+quenching, the amplitude of the Ca2+ influx transient and the resting [ Ca2+] i in PASMCs of CH-exposed and MCT-treated rats. Conclusion Notoginsenoside R1 inhibits SOCE and reduces resting [ Ca2+] i in PASMCs of CH-and MCT-induced PH rats.
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Aim To evaluate the time-course curve of expression of TRPC1 and vascular tone of pulmonary arteries(PAs)mediated by SOCE in chronic hypoxia pulmonary hyperte-nsion rats.Methods Both tension of PA rings and expression of TRPC1 were tested in CH exposure (1 0.0 % ±0.5 %partialpressure ofoxygen ) induced pulmonary hypertensive (PH)rats,and the time-course curve(detected respectively in CH 1 ,3,5, 7,1 4,21 d)was traced.Results ①CH could up-regulate the mean right ventricular pressure(mRVSP) ,which was increased significantly on 1 d,and reached the maximum on 7d;right ventricular weight index (RV-MI)began increase on 3d,and kept rising;②semi-quantitative reverse transcription polyme-rase chain reaction (RT-PCR)was performed to detect the expression of TRPC1 in PAs.The expression of TRPC1 increased significantly on 1 d,and reached the maxi-mum on 3d;③CH could up-regulate the vascular tone of PAs mediated by SOCE,which was increased signif-icantly on 3d,and reached the maximum on 7d.Con-clusions TRPC1 /SOCE increases significantly in the early days of CH,and the time-course curve of the two has correlation,which reflects the important role of the upregulation of TRPC1 /SOCE in the process of chronic hypoxia pulmonary hypertension.
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Objective To investigate the NAD+ - dependent protein deacetylase SIRT1 expression in the cochlea of C57BL/6 mice ,a mouse model of age - related hearing loss(AHL) .Methods A total of 46 C57BL/6 mice were used ,and were divided into 2 groups ;according the age ,there were young group (1 ~ 2 months old ,23 mice) and old group (12 ~ 16 months old ,23 mice) .ABR measurements were performed on young and old C57BL/6 mice . The expression of SIRT1 in the cochlea was detected by qRT - PCR and immunofluorescence .Results The ABR thresholds in the old mice(4 kHz :82 .7 ± 7 .32 dB SPL ,8 kHz :80 .9 ± 7 .8 dB SPL ,16 kHz :89 .3 ± 5 .5 dB SPL ,32 kHz :89 .9 ± 4 dB SPL) were significantly higher than those in the young C57BL/6 mice(4 kHz :52 .1 ± 8 .3 dB SPL ,8 kHz :40 .5 ± 6 .1 dB SPL ,16 kHz :50 .7 ± 9 .4 dB SPL ,32 kHz :57 .6 ± 11 .9 dB SPL)(P < 0 .001) .SIRT1 mRNA expression was significantly decreased in the cochlea of old C57BL/6 mice in comparison with young mice ( P <0 .01) .SIRT1 protein was abundantly expressed in the inner hair cells ,outer hair cells ,supporting cells ,strial marginal cells ,strial intermediated cells ,and spiral ganglion neurons .The positive area and the average flourescence intensity of SIRT1 protein were reduced in old C57BL/6 mice(P< 0 .001) .Conclusion The down - regulation of SIRT1 mRNA and protein expression in the older C57BL/6 mouse cochlea may be correlated with the pathogenesis of AHL .
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Obective To evaluate the effects of dihydrotestosterone (DHT) on the expression of sterol regulatory element-binding protein-1c (SREBP-1c) in human HaCaT keratinocytes.Methods HaCaT cells were cultured in vitro and classified into 4 groups,i.e.,control group receiving no treatment,DIIT group treated with 3 different concentrations (10,100,1000 nmol/L) of DHT,LY294002 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the PI3K inhibitor LY294002 of 50 μmol/L,PD98059 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the MEK inhibitor PD98059 of 50 μmol/L.After another 24-hour culture,real time PCR and Western blot were carried out to detect the expression of SREBP-1c mRNA and protein in HaCaT cells,respectively.Western blot was also performed to determine the phosphorylation levels of protein kinase B (AKT),extracellular signal-regulated kinase (ERK),p38 mitogen activated protein kinase and c-Jun N-terminal kinase (JNK) in the HaCaT cells.Results DHT could enhance the expression of SREBP-1c mRNA and protein in HaCaT cells in a concentration-dependent manner,and induce the phosphorylation of AKT and ERK,but not that of P38 or JNK.The expressions of SREBP-1c mRNA and protein were significantly decreased in HaCaT cells treated with LY294002 plus DHT (7.4780 ± 1.2638 vs.21.6170 ± 2.2759,t =9.406,P ＜ 0.05; 0.7113 + 0.0313 vs.2.2577 + 0.0601,t =39.498,P ＜ 0.05),but experienced no statistical changes in those treated with PD98059 and DHT(both P ＞ 0.05),compared with those treated with DHT only.Conclusion DHT can induce the expression of SREBP-1c mRNA and protein in HaCaT cells,likely via the PI3K/AKT signaling pathway.
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OBJECTIVE@#Comparison of the sensitivity of Sp/Ap amplitude and area ratio in the aid of diagnosis of Meniere's disease. Meniere's disease was defined by the 1995 American Academy of Otolaryngology-Head and Neck Surgery Committee on Hearing and Equilibrium.@*METHOD@#The study sample consisted of 35 patients in our hospital, who were undergone tympanic electrocochleography (EcochG). A control group of 10 patients (20 ears) with normal Sp/Ap ratio and EcochG data were identified. Sp/Ap amplitude and area curve ratios for both group were measured.@*RESULT@#The traditional upper limit and the upper limit of normal for Sp/Ap amplitude and area curve were respectively 0.4, 46 and 1.94. Of 35 patients with Meniere's disease, had an abnormal Sp/Ap amplitude ratio,and had an abnormal Sp/Ap area curve ratio; the difference between groups was statistically significant.@*CONCLUSION@#Sp/Ap curve area curve ratio is more sensitive than conventional Sp/Ap amplitude ratios in diagnosis of Meniere's disease.
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Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Audiometry, Evoked Response , Case-Control Studies , Endolymph , Meniere Disease , Diagnosis , Sensitivity and SpecificityABSTRACT
ObjectiveTo observe the effect of topical sulfotanshinone sodium(STS) on sebaceous hyperplasia in animal models. MethodsThe sebaceous gland spots of adult male Syrian hamster flank organ served as the animal model. Sulfotanshinone sodium(0.5％) was applied to sebaceous gland spots in the right flank organ thrice daily, while those in the left were treated with normal saline as control. Parameters were examinedbefore, 10 days, 20 days and 30 days after the beginning of the topical treatment. A vernier caliper was utilized to measure the size of sebaceous gland spots, hematoxylin and eosin(HE) staining to observe the structure of sebaceous glands, immunohistochemistry to determine the expression of proliferating cell nuclear antigen (PCNA) in sebaceous gland cells, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to assess the apoptosis of sebaceous gland cells. ResultsAt the baseline, no significant difference was observed in the size of sebaceous gland spots or in the proliferation and apoptosis of sebaceous gland cells between the two sides of flank organ(all P ＞ 0.05), with tightly arranged intact sebaceous glands. Compared with normal saline, sulfotanshinone sodium significantly reduced the size of sebaceous gland spots(P ＜ 0.05). Sebaceous glands were loosely arranged with decreased quantity and volume and obviously atrophic on day 30 in the right flank organ of hamsters. A decrease was observed in the expression of PCNA in sulfotanshinone sodium treated sebaceous gland cells compared with those treated with normal saline(P ＜ 0.01 ), which was more striking on day 10 and 20(both P ＜ 0.005). Sulfotanshinone sodium also induced an enhancement of apoptosis in sebaceous gland cells (P ＜ 0.01 ), which was more apparent on day 20 (P ＜ 0.005 ), and the degree of apoptosis was higher in the central area than in the peripheral area of sebaceous glands. ConclusionSulfotanshinone sodium can reduce the size and alter the microstructure of sebaceous gland spots, and inhibit the hyperplasia of sebaceous glands.