1.Mechanism of Modified Danggui Shaoyaosan in Improving Inflammation and Apoptosis in Acne via Regulating JNK/p38 MAPK Pathway
Gongzhen CHEN ; Yuqi YANG ; Xin LIU ; Ting TANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):31-40
ObjectiveTo explore the therapeutic effects and mechanisms of modified Danggui Shaoyaosan on acne based on the c-Jun N-terminal kinase (JNK)/p38 mitogen-activated protein kinase (p38 MAPK) pathway. MethodsA rat ear acne model was established in SD rats, and the rats were divided into a blank group, a model group, and low-, medium-, and high-dose groups of modified Danggui Shaoyaosan (7.15, 14.30, 28.60 g·kg·d-1), with six rats in each group. After the administration for 14 consecutive days, morphological changes in the rats' auricles were observed, and hematoxylin-eosin (HE) staining was used to examine the pathological changes in the acne-affected ear tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the ear tissue. Quantitative reverse transcription polymerase chain reaction (Real-time PCR) was performed to detect the mRNA expression levels of Caspase-3, B cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), JNK, and p38 MAPK in the ear tissue. Additionally, Western blot analysis was conducted to assess the protein levels of Caspase-3, Bcl-2, Bax, JNK, and p38 MAPK in the ear tissue. The active components and key targets of modified Danggui Shaoyaosan in the treatment of acne were identified through network pharmacology analysis. Molecular docking was then employed to evaluate the interactions between the main active components and the key targets. ResultsThe results of the animal experiment demonstrated that compared with those in the blank group, rats in the model group exhibited redness, swelling, thickening, hardening, dryness, and roughness of the auricle. The surface showed sebaceous scales and desquamation, accompanied by acne-like lesions such as papule-like elevations or cysts. Histopathological changes included keratinization, epidermal thickening, dermal collagen fiber degeneration and necrosis, subcutaneous muscle degeneration and necrosis, inflammatory cell infiltration, and fibrous tissue proliferation. The mRNA and protein expression levels of IL-1β, TNF-α, Caspase-3, Bax, JNK, and p38 MAPK were significantly increased (P<0.01), while those of Bcl-2 were significantly decreased (P<0.01). In comparison to the model group, the modified Danggui Shaoyaosan groups showed marked improvement in acne-like lesions of the auricle, with varying degrees of histopathological damage reduction. Additionally, the mRNA and protein expression levels of IL-1β, TNF-α, Caspase-3, Bax, JNK, and p38 MAPK in the tissue were significantly decreased (P<0.05, P<0.01), while those of Bcl-2 were significantly increased (P<0.05, P<0.01). Network pharmacology analysis indicated that the key compounds in modified Danggui Shaoyaosan responsible for its effects in treating acne may include acacetin, kaempferol, luteolin, quercetin, wogonin, and baicalein. These compounds exerted their effects by modulating core targets such as TNF, IL-1β, Caspase-3, and Bcl-2, thereby alleviating inflammation and apoptosis and ultimately improving acne symptoms. ConclusionModified Danggui Shaoyaosan may exert its therapeutic effects on acne by inhibiting the activation of the JNK/p38 MAPK pathway, thereby alleviating inflammation and apoptosis.
2.Study on the inhibitory effect and mechanism of Modified qifang weitong granules on gastric cancer
Xinyuan CHEN ; Chengting WU ; Changzhou XIONG ; Ting WANG ; Yinhang CUI ; Peibin WU ; Wenlong CHEN ; Huilin CHEN ; Caizhi LIN ; Meiwen TANG
China Pharmacy 2025;36(21):2656-2661
OBJECTIVE To investigate the inhibitory effect and mechanism of Modified qifang weitong granules on gastric cancer based on in vitro and in vivo experiments. METHODS Human gastric cancer HGC-27 cells were divided into the following groups: control group (treated with fetal bovine serum), 10% drug-containing serum group, 15% drug-containing serum group, 20% drug-containing serum group, and 5-fluorouracil (5-Fu) group (positive control, 3.90 μg/mL). After culturing the cells in each group with the corresponding serum/drug solution, their proliferation, migratory and invasive abilities, as well as the cell cycle, were assessed. Additionally, the expression levels of epithelial-mesenchymal transition (EMT)-related proteins [E-cadherin, N-cadherin, and vimentin] in the cells were measured. Logarithmic-phase HGC-27 cells were harvested and subcutaneously injected into the right axillary region of nude mice to establish a subcutaneous xenograft tumor model in nude mice. The successfully modeled tumor-bearing nude mice were randomly divided into model group, low-, medium- and high-dose groups of Modified qifang weitong granules (17.65, 35.29 and 70.58 g/kg, respectively), and 5-Fu group (25 mg/kg), with 5 mice in each group. After 14 days of treatment with the corresponding drugs in each group, the histopathological morphology of the tumor tissues in the nude mice was observed. Immunohistochemistry and Western blot assay were employed to detect the expression levels of EMT- related proteins in the tumor tissues of the nude mice. RESULTS In the cell experiment, compared with the control group, the cell proliferation rate, migration rate, number of invasive cells, as well as the expression levels of N-cadherin and vimentin proteins, and the percentage of cells in the G2/M phase were all significantly decreased/reduced in the 15% drug-containing serum group, 20% drug-containing serum group (P<0.05). Conversely, the percentage of cells in the G0/G1 phase and the expression level of E- cadherin protein were significantly increased (P<0.05). In animal experiment, compared with the model group, the high-dose group of Modified qifang weitong granules exhibited significantly reduced tumor mass and expression levels of N-cadherin and vimentin proteins in the tumor tissues of nude mice (P<0.05), while the expression level of E-cadherinprotein in the tumor tissues was significantly increased (P<0.05). Additionally, the tumor cells varied in size and showed extensive necrosis. CONCLUSIONS Modified qifang weitong granules effectively inhibit gastric cancer in both in vitro and in vivo models, and the mechanism of action is related to the suppression of EMT.
3.Cervical spondylosis: innovative understanding from traditional Chinese medicine and treatment by classic formulas.
Heng CHEN ; Cong-Yang XUE ; Shuang CHEN ; Zi-Ting CHEN ; Tian TANG ; Xin LIU ; Zhi-Peng XI ; Ran KANG ; Lin XIE
China Journal of Chinese Materia Medica 2025;50(9):2596-2604
As one of the chronic diseases with high incidence in contemporary society, cervical spondylosis has increasing patient groups who gradually present a low age, and it seriously affects social and public health. Although modern medicine has made great progress in the pathological research and clinical treatment of cervical spondylosis, patients still face gastrointestinal side effects of nonsteroidal anti-inflammatory drugs(NSAIDs), neck pain, limited mobility, upper limb numbness, and other symptoms after conservative or surgical treatment. In the theory of traditional Chinese medicine(TCM), cervical spondylosis belongs to the categories of "Bi syndrome" "stiff neck" "stiff Bi", etc. With the change of the times, the change of lifestyle, and the application of western medicine treatment, the etiology and pathogenesis of TCM in cervical spondylosis also show new characteristics. In terms of etiology and pathogenesis, it involves the invasion of wind, cold, and dampness, long-term strain, liver and kidney deficiency, Qi and blood stasis, which are associated with factors such as cervical degeneration, muscle tension and spasm, intervertebral disc herniation, and nerve root compression in modern medicine. In terms of the evolution of pathogenesis, in the early stage, wind, cold, and dampness, were more common in Xuanfu, resulting in unfavorable muscles and bones, poor flow of Qi and blood, and cervical spondylosis and radiculopathy. Medium-term phlegm stasis and internal knots, sluggish muscles and veins, and long-term weathering and fire are more likely to occur in the vertebral artery and sympathetic radiculopathy. In the later stage, the positive Qi is depleted; the true Yin is damaged, and the viscera Qi and blood are deficient, which is most common in cervical myelopathy. The strategy of treating cervical spondylosis with TCM classic formulas applies Gegen Decoction, Wutou Decoction, Qianghuo Shengshi Decoction, Mahuang Jiazhu Decoction to patients with wind, cold, and dampness. Patients with phlegm dampness and blood stasis are treated with Huoxue Xiaoling Dan, Jinlingzi Powder, Siwu Decoction, Banxia Baizhu Tianma Decoction, Shuanghe Decoction, etc. For those patients with liver, spleen, and kidney deficiency, Huangqi Guizhi Wuwu Decoction, Tianma Gouteng Decoction, Guishao Dihuang Pills, Shenling Baizhu Powder, and Lizhong Decoction are used to invigorate the spleen, nourish Qi and blood, and tonify liver and kidney. In clinical practice, the authors advocate a safe and effective treatment plan of classic formulas based on deficiency and excess, the integration of formulas and syndromes, and the combination of modern research results, so as to relieve symptoms, reduce recurrence, and reduce medical burden.
Humans
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Spondylosis/drug therapy*
;
Medicine, Chinese Traditional/methods*
;
Drugs, Chinese Herbal/therapeutic use*
;
Cervical Vertebrae/pathology*
4.Two cases of creatine deficiency syndrome caused by GAMT gene mutations and literature review.
Ting-Ting ZHAO ; Zou PAN ; Jian-Min ZHONG ; Hai-Yun TANG ; Fei YIN ; Jing PENG ; Chen CHEN
Chinese Journal of Contemporary Pediatrics 2025;27(3):340-346
OBJECTIVES:
To summarize the clinical manifestations and genetic characteristics of creatine deficiency syndrome (CDS) caused by GAMT gene mutations.
METHODS:
A retrospective analysis was conducted on the clinical and genetic data of two children diagnosed with GAMT deficiency-type CDS at the Children's Medical Center of Xiangya Hospital, Central South University, from December 2020 to December 2024.
RESULTS:
The two patients presented with symptoms in infancy, and both had compound heterozygous mutations in the GAMT gene. Case 1 exhibited seizures and intellectual disability, while Case 2 had intellectual disability and attention-deficit hyperactivity disorder. Magnetic resonance spectroscopy of cranial MRI in both patients indicated reduced creatine peaks. After creatine treatment, seizures in Case 1 were controlled, but both patients continued to experience intellectual disabilities and behavioral issues. As of December 2024, a total of 21 cases have been reported in China (including this study), and 115 cases have been reported abroad. All patients exhibited developmental delay or intellectual disabilities, with 66.9% (91/136) experiencing seizures, 33.8% (46/136) presenting with motor disorders, and 36.8% (50/136) having behavioral problems. Seventy-five percent (102/136) of patients received creatine treatment, leading to significant improvements in seizures and motor disorders, although cognitive improvement was not substantial.
CONCLUSIONS
GAMT deficiency-type CDS is rare and presents with nonspecific clinical features. Timely diagnosis facilitates targeted treatment, which can partially improve prognosis.
Child
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Female
;
Humans
;
Male
;
Creatine/deficiency*
;
Guanidinoacetate N-Methyltransferase/deficiency*
;
Intellectual Disability/genetics*
;
Mutation
;
Retrospective Studies
;
Rhabdomyolysis/genetics*
;
Language Development Disorders
;
Movement Disorders/congenital*
5.Chain mediating role of family care and emotional management between social support and anxiety in primary school students.
Zhan-Wen LI ; Jian-Hui WEI ; Ke-Bin CHEN ; Xiao-Rui RUAN ; Yu-Ting WEN ; Cheng-Lu ZHOU ; Jia-Peng TANG ; Ting-Ting WANG ; Ya-Qing TAN ; Jia-Bi QIN
Chinese Journal of Contemporary Pediatrics 2025;27(10):1176-1184
OBJECTIVES:
To investigate the chain mediating role of family care and emotional management in the relationship between social support and anxiety among rural primary school students.
METHODS:
A questionnaire survey was conducted among students in grades 4 to 6 from four counties in Hunan Province. Data were collected using the Social Support Rating Scale, Family Care Index Scale, Emotional Intelligence Scale, and Generalized Anxiety Disorder -7. Logistic regression analysis was used to explore the influencing factors of anxiety symptoms. Mediation analysis was conducted to assess the chain mediating effects of family care and emotional management between social support and anxiety.
RESULTS:
A total of 4 141 questionnaires were distributed, with 3 874 valid responses (effective response rate: 93.55%). The prevalence rate of anxiety symptoms among these students was 9.32% (95%CI: 8.40%-10.23%). Significant differences were observed in the prevalence rates of anxiety symptoms among groups with different levels of social support, family functioning, and emotional management ability (P<0.05). The total indirect effect of social support on anxiety symptoms via family care and emotional management was significant (β=-0.137, 95%CI: -0.167 to -0.109), and the direct effect of social support on anxiety symptoms remained significant (P<0.05). Family care and emotional management served as significant chain mediators in the relationship between social support and anxiety symptoms (β=-0.025,95%CI:-0.032 to -0.018), accounting for 14.5% of the total effect.
CONCLUSIONS
Social support can directly affect anxiety symptoms among rural primary school students and can also indirectly influence anxiety symptoms through the chain mediating effects of family care and emotional management. These findings provide scientific evidence for the prevention of anxiety in primary school students from multiple perspectives.
Humans
;
Female
;
Male
;
Social Support
;
Anxiety/etiology*
;
Child
;
Students/psychology*
;
Emotions
;
Logistic Models
6.The Sequential Mediating Roles of Body Pain and Self-Reported Health Status in the Relationship between Sleep Duration and Life Satisfaction.
Jia Feng LI ; Xue Wei FU ; Dan YANG ; Ye WANG ; Ting CHEN ; Yang PENG ; Feng Hao YANG ; Yu Chen ZHAN ; Yu WANG ; Xiang Dong TANG
Biomedical and Environmental Sciences 2025;38(1):47-55
OBJECTIVE:
This study examines the sequential mediating roles of body pain and self-reported health in the association between sleep duration and self-reported life satisfaction among elderly Chinese adults.
METHODS:
Data from the fifth wave of the China Health and Retirement Longitudinal Survey (CHARLS) were used to analyse the relationships between sleep duration and body pain, self-reported health, and life satisfaction through logistic regression and Restricted Cubic Spline (RCS) analyses. The sequential mediation effects of body pain and self-reported health status were examined via chain mediation analysis.
RESULTS:
Logistic regression analysis showed that sleeping fewer than 6 hours or 6-7 hours was linked to higher risks of body pain, poor health, and dissatisfaction with life compared to sleeping 7-8 hours (all P < 0.05). Additionally, those sleeping more than 9 hours also had increased risks of poor health and dissatisfaction with life compared to those sleeping 7-8 hours (all P < 0.05). Chain mediation analysis showed that body pain and self-reported health status sequentially mediated 46.15% of the association between sleep duration and life satisfaction.
CONCLUSION
Body pain and self-reported health may shape the relationship between sleep duration and life satisfaction in elderly Chinese adults.
Humans
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Male
;
Female
;
Aged
;
Personal Satisfaction
;
Sleep
;
Health Status
;
Self Report
;
China
;
Middle Aged
;
Longitudinal Studies
;
Pain/psychology*
;
Sleep Duration
7.Epidemiological characteristics of rifampicin resistant pulmonary tuberculosis among students in Chongqing during 2015-2024
Chinese Journal of School Health 2025;46(12):1771-1775
Objective:
To delineate the epidemiologic profile of rifampicin resistant pulmonary tuberculosis (RR-PTB) among students in Chongqing, so as to provide evidence for effectively controlling RR-PTB outbreaks in schools.
Methods:
Individual level surveillance records of 395 student RR-PTB cases reported from 2015 to 2024 were extracted from the China Information System for Disease Control and Prevention. The Joinpoint regression analysis was employed to quantify temporal trends in the registration rate of student RR-PTB cases, and the comparison of RR-PTB registration rates with different demographic characteristics and different regions was performed using Chi-square test.
Results:
From 2015 to 2024, a total of 395 student RR-PTB cases were identified, with the registration rate ranged from 0.07 per 100 000 to 1.47 per 100 000, showed a fluctuating upward trend ( AAPC= 35.22%, t =4.13, P <0.01). A turning point was detected in 2017, rates rose during 2015-2017 (APC=295.23%, t =4.62, P < 0.01 ) and plateaued thereafter (APC=-0.47%, t =-0.12, P =0.91). The proportion of RR-PTB cases occurring among students increased both among all RR-PTB cases (1.54% in 2015, 7.48% in 2024) and all student pulmonary tuberculosis cases (0.20% in 2015, 7.17% in 2024), with significant linear trends ( χ 2 trend =33.55,159.98, both P <0.01). The majority of cases were enrolled in senior high school (50.38%), classified as retreatment (53.92%), of Han ethnicity (75.95%), and diagnosed with multidrug resistant tuberculosis(53.16%). There were significant differences in the composition of different ethnicity, registration category and resistance pattern between different years( χ 2=23.47, 17.23, 59.64,all P <0.05). The South-Eastern Wuling Mountainous Region exhibited the highest notification rate (3.96 per 100 000), whereas the western region had the lowest rate ( 0.47 per 100 000). County level jurisdictions reported higher rates than district level ones (2.16 per 100 000 vs 0.63 per 100 000 ). Statistically significant differences were observed in the RR-PTB reported rates among students across different districts and counties( χ 2=418.05,167.05,both P <0.01).
Conclusions
From 2015 to 2024, the registration rate of detected student RR-PTB cases in Chongqing showed an increasing trend. Students have become one of the key populations for drug resistant TB prevention and control. Intensified health education and active case finding should be implemented to enhance proactive surveillance capabilities.
8.Curative effect of Shuotong ureteroscopy combined with flexible ureteroscope in the treatment of 2-3 cm lower calyceal calculi
Yilong LIANG ; Ting DENG ; Shen CHEN ; Ziyan TANG ; Zhen LI ; Guoyou HE ; Jinsong PANG
International Journal of Surgery 2024;51(2):77-85
Objective:To investigate the efficacy of Shuotong ureteroscope combined with flexible ureteroscope in the treatment of 2-3 cm lower calyceal calculi, and analyze the influencing factors.Methods:A total of 102 patients with lower calyceal calculi were treated in the Second People′s Hospital of Yulin from February 2019 to December 2022, and they were divided into the observation group and the control group, with 51 cases in each group. The patients of the observation group were treated with Shuotong ureteroscope combined with flexible ureteroscope, while the patients of the control group were treated with flexible ureteroscope. According to whether the stones were completely removed after operation, all patients were divided into non-stone removal group ( n=13) and stone removal group ( n=89). The operation time, hospitalization time, lithotripsy time, intraoperative blood loss, complication rate and stone clearance rate were compared between the observation group and the control group. Generalized Estimation Equation was used to analyze and evaluate the effects of treatment time, treatment scheme and their interaction on visual analogue scale (VAS), white blood cell (WBC), blood urea nitrogen (BUN), blood creatinine (Cr), hemoglobin (HGB) and procalcitonin (PCT). Univariate and multivariate Logistic regression were used to analyze the risk factors of stone removal rate. Nomogram model was constructed based on risk factors and evaluate the model. Results:Compared with the control group, operation time [(118.72±9.61) min vs (136.65±11.27) min], hospitalization stay [(6.43±1.12) d vs (10.29±2.23) d] and the lithotripsy time [ (51.23±10.38) min vs (56.62±11.43) min] of the observation group were shorter, and the amount of intraoperative blood loss [(128.52±10.20) mL vs (157.53±15.31) mL] were significantly less than those of the control group ( P< 0.05). The results of Generalized Estimation Equation analysis showed that treatment time, treatment regimen and their interaction had significant effects on WBC, HGB, BUN, Cr, PCT and VAS ( P< 0.05). Compared with the control group, the incidence of complications (5.88% vs 19.61%) of the observation group was lower and the stone clearance rate ( 94.12% vs 80.39%) was significantly higher than those in the control group ( P< 0.05). The mode of operation, infundibulopelvic angle(IPA), caliceal pelvic height (CPH) and the maximum diameter of stones were all influencing factors of stone removal rate in patients with 2-3 cm lower calyceal calculi. The nomogram model constructed in this study has good differentiation, calibration and clinical practicability, and can better identify high-risk patients with incomplete removal of 2-3 cm lower calyceal calculi. Conclusions:Shuotong ureteroscope combined with flexible ureteroscope is a safe, effective method for the treatment of 2-3 cm lower calyceal calculi. It has the advantages of simple operation, less intraoperative bleeding, less postoperative complications and high stone clearance rate. IPA, CPH, the maximum diameter of calculi and the mode of operation were all independent factors affecting the stone clearance rate of 2-3 cm lower calyceal calculi. The nomogram model constructed in this study can well identify the high-risk patients with incomplete clearance of 2-3 cm lower calyceal calculi.
9.The Role of Cerebral-Placenta-Uterine Ratio in Predicting Late-Onset Fetal Growth Restriction
Yongyan CHU ; Haiyan TANG ; Jiayi ZHANG ; Chuqin XIONG ; Haoyue HUANG ; Runhe LIANG ; Cuiying LEI ; Ting ZENG ; Yanyan LI ; Li HE ; Minping CHEN ; Libei DU ; Shengmou LIN
Journal of Practical Obstetrics and Gynecology 2024;40(1):36-41
Objective:To evaluate the efficacy of cerebral-placental-uterine ratio(CPUR)in predicting late-on-set fetal growth restriction(FGR).Methods:From May 2020 to May 2021,1255 women with singleton pregnancy who underwent prenatal examinations at the University of Hong Kong Shenzhen Hospital were selected for fetal growth and Doppler measurements at 35-37 +6 weeks of gestation.Pregnant women with birth weight of newbo-rns<the 10th percentile were the FGR group.The pulsatility index(PI)of uterine artery(UtA),umbilical artery(UA)and fetal middle cerebral artery(MCA)were analyzed separately and in combination.ROC curve was used to analyze the cerebral-placental-uterine ratio(CPUR),cerebral-placental ratio(CPR),cerebral-uterine ratio(C-UtA)for predicting late-onset FGR;and to evaluate the sensitivity,positive and negative predictive value and of CPUR in the prediction of late-onset FGR.Results:The area under the curve(AUC)of CPUR,CPR,C-UtA and mean UtA-PI for FGR grope were 0.88,0.86,0.84 and 0.72.Under certain cut-off values and 87% specificity,the specificity of CPUR,CPR,C-UtA and mean UtA-Pifor predicting FGR group was 43.2%,46.6%,39.8% and 23.9%,respectively.The positive predictive values of CPUR,CPR,C-UtA and mean UtA-PI,UA-PI for predicting FGR group were 90.5%,71.9%,83.3%,63.6%and 5.2%,respectively.Conclusions:CPUR is more effective in predicting late onset FGR than CPR,C-UtA and mean UtA-PI.It can effectively increase the detection rate of fetal growth restrictionand reduce the FGR risk.
10.Effect of sleep deprivation on expression of SIRT6 in cerebellum of immature mice
Lan XIAO ; Chenyi GAO ; Ting CHEN ; Xuemin ZHANG ; Qi ZHONG ; Yue CAO ; Lijuan TANG ; Chang CHEN ; Zongze ZHANG
Chinese Journal of Anesthesiology 2024;44(1):41-45
Objective:To evaluate the effect of sleep deprivation on the expression of sirtuin 6 (SIRT6) in the cerebellum of immature mice.Methods:Fifty SPF healthy male C57BL/6 mice, aged 4 weeks, weighing 14-16 g, were divided into 2 groups ( n=25 each) using a random number table method: control group (Con group) and sleep deprivation group (SD group). The chronic sleep deprivation model was prepared by using the multi-platform water environment method, with 20 h of sleep deprivation per day for 10 consecutive days. After sleep deprivation, a balance beam experiment was performed to test the balance and coordination ability of mice. The mice were sacrificed after anesthesia and cerebellar lobular IV-VI (4-6 cb) tissues were taken for microscopic examination of the ultrastructure (with a transmission electron microscope) and for determination of the dendritic spine density of cerebellar 4-6cb Purkinje neurons (by Golgi staining), co-expression of SIRT6 and Calbindin D-28k (CbD-28k) and expression of glucose transporter Glut3 of cerebellar 4-6cb (by immunofluorescence staining). Results:Compared with group Con, the duration of passage through the balance beam was significantly prolonged, and the number of posterior foot slips was increased, the synaptic gap of cerebellar 4-6cb neurons was increased, the thickness of postsynaptic density was increased, the density of dendritic spines of Purkinje cells and the number of positive cells co-expressing SIRT6 and CbD-28k were decreased, and the expression of Glut3 was down-regulated in group SD ( P<0.05). Conclusions:The mechanism by which sleep deprivation decreases the abilities of balance and coordination is related to down-regulating SIRT6 expression in cerebellar Purkinje cells and decreasing neuronal glucose metabolism, thus damaging the synaptic plasticity of cerebellum in immature mice.


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