1.Mechanism of IGF2BP2 regulation of PPAR-γ/GLUT4 pathway in insulin resistance induced by sodium arsenite exposure in HepG2 cells
Shiqing XU ; Zhida HU ; Qiyao ZHANG ; Siqi ZHAO ; Yujie WANG ; Xiaohui WANG ; Teng MA ; Li WANG
Journal of Environmental and Occupational Medicine 2025;42(4):400-407
Background Arsenic is an environmentally harmful substance that causes hepatic insulin resistance and liver damage, increasing the risk of type 2 diabetes mellitus. Objective To explore whether the insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) is involved in insulin resistance in HepG2 cells after arsenic exposure through the peroxisome-proliferator-activated receptor γ (PPAR-γ) / glucose transporter 4 (GLUT4) pathway. Methods Cell viability was determined using cell counting kit 8 (CCK8) and an appropriate NaAsO2 infection dose was determined. A cellular arsenic exposure model of HepG2 cells was established by four concentrations of NaAsO2 solution for 24 h (the experiment was divided into four groups: 0, 2, 4, and 8 μmol·L−1); HepG2 cells were firstly treated with pcDNA3.1-IGF2BP2 and pcDNA3.1-NC respectively for 6 h, then with 8 μmol·L−1 NaAsO2 for 24 h to establish a IGF2BP2 overexpression cell model (the experiment was divided into 4 groups: control, NaAsO2, NaAsO2+pcDNA3.1-IGF2BP2, and NaAsO2+pcDNA3.1-NC); finally the cells were subject to 100 nmol·L−1 insulin stimulation for 30 min. Glycogen and glucose in HepG2 cells were determined by glycogen and glucose assay kits; mRNA expression levels of IGF2BP2 were measured by quantitative real-time PCR; protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in HepG2 were detected by Western blot (WB); and the binding of IGF2BP2 to PPAR-γ and PPAR-γ to GLUT4 was verified by co-immunoprecipitation (CO-IP) experiment. Results The results of CCK8 experiment showed a dose-effect relationship between NaAsO2 concentration and cell viability. When the concentration of NaAsO2 was ≥4 μmol·L−1 , the cell viabilities were lower than that of the control group (P <0.05). With the increasing dose of NaAsO2 infection, reduced glucose consumption and glycogen levels in HepG2 cells were found in the 2, 4, and 8 μmol·L−1 NaAsO2 treatment groups compared to the control group (P <0.05). The difference between the mRNA expression level of IGF2BP2 in the HepG2 cells treated with 4 or 8 μmol L−1 NaAsO2 and the control group was significant (P <0.05). In the IGF2BP2 overexpression cell model, compared with the control group, glucose consumption and glycogen levels were lowered in the NaAsO2 group (P <0.05), the mRNA expression level of IGF2BP2 and the protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in the cell membrane were all decreased (P <0.05). Compared with the NaAsO2 group, the glucose consumption and glycogen levels were increased in the NaAsO2+pcDNA3.1-IGF2BP2 group (P <0.05), and the mRNA expression level of IGF2BP2 and the protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in the cell membrane were all increased (P <0.05). The results of CO-IP experiments showed that IGF2BP2 interacted with PPAR-γ as well as PPAR-γ with GLUT4 protein. Conclusion IGF2BP2 is involved in arsenic exposure-induced insulin resistance in HepG2 cells by acting on the PPAR-γ/GLUT4 pathway.
2.Research progress on the mechanism of traditional Chinese medicine monomers acting on myelosuppression after chemotherapy based on the four properties theory
Sihan ZHANG ; Tingting WANG ; Zhifen ZHAO ; Hanyu KANG ; Jiaqi JI ; Ziqiang GUO ; Tong LIU ; Shiqing JIANG
China Pharmacy 2025;36(18):2341-2347
Chemotherapy is an important treatment for tumors, but most patients experience varying degrees of chemotherapy- induced myelosuppression. Four properties theory of traditional Chinese medicine (TCM) has unique advantages in improving chemotherapy-induced myelosuppression. The monomers from TCM with different properties and flavors, such as cold-natured (e.g. Scutellaria baicalensis, Rhus chinensis), cool-natured (e.g. Ligustrum lucidum, Ophiopogon japonicus), warm-natured (e.g. Panax ginseng, Epimedium brevicornu, Curcuma longa, Angelica sinensis), hot-natured (e.g. Cinnamomum cassia, Aconitum carmichaeli), and neutral-natured (e. g. donkey-hide gelatin, Lycium barbarum, Rhodiola rosea, fungi), can exert anti- myelosuppressive effects by reducing damage to hematopoietic stem/progenitor cells, improving the bone marrow hematopoietic microenvironment, inhibiting the oxidative stress response, regulating signaling pathways, so as to ultimately repaire inflammatory damage and improve hematopoietic function, thereby playing an anti-myelosuppressive role.
3.Regulation of Immune Balance by Traditional Chinese Medicine in Treatment of Cough Variant Asthma: A Review
Han YANG ; Yonghuang YAN ; Wenting ZHANG ; Peixuan ZHU ; Fang YAN ; Yujie WU ; Shiqing QIAO ; Tieshan WANG ; Zeqi SU ; Ting WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):206-212
Cough variant asthma (CVA) is a chronic respiratory disease with cough as its main symptom. The occurrence of CVA is closely related to non-specific airway inflammation, and its pathogenesis involves environmental, genetic, immune, and other factors. In recent years, the advantages of traditional Chinese medicine (TCM) in the treatment of CVA have attracted the attention of experts and scholars in China and abroad, especially its prominent role in regulating immune balance, relieving cough symptoms in CVA patients, and reducing recurrence. T Helper cells 1 (Th1), T helper cells 2 (Th2), T helper cells 17 (Th17), and regulatory T cells (Treg) are derived from CD4+ T cells. Immune imbalance of Th1/Th2 and Th17/Treg is a new hotspot in the pathogenesis of CVA and a potential key target in the treatment of CVA by TCM. Th cell subsets are in dynamic balance under physiological conditions, maintaining respiratory immune homeostasis in which pro-inflammatory cytokines and anti-inflammatory cytokines are balanced. Immature helper T cells (Th0) can be differentiated into Th1, Th2, Th17, Treg, and other cell subsets due to cytokine types in the microenvironment in the stage of CVA maturation. The proliferation of Th2 cells leads to eosinophilic airway inflammation. Excessive differentiation of Th17 cells induces neutrophil airway inflammation. Th1/Th2 and Th17/Treg cells are mutually restricted in number and function, and the immune imbalance of Th1/Th2 and Th17/Treg is easy to aggravate the generation of inflammatory response. Restoring immune balance is particularly important for the airway anti-inflammatory therapy of CVA. In this paper, the imbalance of Th1/Th2 and Th17/Treg and the pathogenesis of CVA were systematically expounded. Meanwhile, the latest research on the regulation of immune imbalance by TCM compound, single TCM, and its effective ingredients in the treatment of CVA was reviewed. It provides ideas and references for revealing the scientific connotation of TCM regulating immune balance therapy of CVA, as well as the development of clinical treatment and basic research of CVA.
4.Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults (version 2024)
Qingde WANG ; Yuan HE ; Bohua CHEN ; Tongwei CHU ; Jinpeng DU ; Jian DONG ; Haoyu FENG ; Shunwu FAN ; Shiqing FENG ; Yanzheng GAO ; Zhong GUAN ; Hua GUO ; Yong HAI ; Lijun HE ; Dianming JIANG ; Jianyuan JIANG ; Bin LIN ; Bin LIU ; Baoge LIU ; Chunde LI ; Fang LI ; Feng LI ; Guohua LYU ; Li LI ; Qi LIAO ; Weishi LI ; Xiaoguang LIU ; Hongjian LIU ; Yong LIU ; Zhongjun LIU ; Shibao LU ; Yong QIU ; Limin RONG ; Yong SHEN ; Huiyong SHEN ; Jun SHU ; Yueming SONG ; Tiansheng SUN ; Yan WANG ; Zhe WANG ; Zheng WANG ; Hong XIA ; Guoyong YIN ; Jinglong YAN ; Wen YUAN ; Zhaoming YE ; Jie ZHAO ; Jianguo ZHANG ; Yue ZHU ; Yingjie ZHOU ; Zhongmin ZHANG ; Wei MEI ; Dingjun HAO ; Baorong HE
Chinese Journal of Trauma 2024;40(2):97-106
Ankylosing spondylitis (AS) combined with lower cervical fracture is often categorized into unstable fracture, with a high incidence of neurological injury and a high rate of disability and morbidity. As factors such as shoulder occlusion may affect the accuracy of X-ray imaging diagnosis, it is often easily misdiagnosed at the primary diagnosis. Non-operative treatment has complications such as bone nonunion and the possibility of secondary neurological damage, while the timing, access and choice of surgical treatment are still controversial. Currently, there are no clinical practice guidelines for the treatment of AS combined with lower cervical fracture with or without dislocation. To this end, the Spinal Trauma Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults ( version 2024) in accordance with the principles of evidence-based medicine, scientificity and practicality, in which 11 recommendations were put forward in terms of the diagnosis, imaging evaluation, typing and treatment, etc, to provide guidance for the diagnosis and treatment of AS combined with lower cervical fracture.
5.Improvement effect of Phellodendron amurense polysaccharides on gouty nephropathy in rats and its mechanism
Yongzhe MA ; Yuliang WANG ; Kai ZHANG ; Hong ZHAO ; Yu SHEN ; Hongbin QIU ; Chaoxing WANG ; Shiqing SUN ; Zhenxu JIANG ; Mingming SONG ; Yu ZHANG
China Pharmacy 2024;35(5):555-559
OBJECTIVE To study the effects of Phellodendron amurense polysaccharides (PAP) on improving gouty nephropathy (GN) in rats, and to investigate its mechanism primarily by interfering the p38 mitogen-activated protein kinase (p38 MAPK)/nuclear factor-κB(NF-κB)/tumor necrosis factor-α(TNF-α). METHODS Sixty rats were randomly divided into normal group (water), model group (water), allopurinol group (positive control, 20 mg/kg), PAP high-dose, medium-dose and low-dose groups (100, 50, 25 mg/kg, by raw material) after being stratified by body weight, with 10 rats in each group. Except for the normal group, the other groups were induced to construct GN model by giving 1 500 mg/kg potassium oxazinate and 100 mg/kg adenine intragastrically for 14 days. After modeling, the rats in each group were given relevant medicine/water intragastrically, once a day, for consecutive 28 days. After the last medication, the levels of biochemical parameters related to renal function [uric acid, creatinine (Cr), blood urea nitrogen (BUN), xanthine oxidase (XOD)] were detected in rats, and the histopathological changes in the rat kidney were observed. The protein expressions of monocyte chemoattractant protein-1(MCP-1),TNF-α and interleukin-6(IL-6) as well as the phosphorylation levels of p38 MAPK and NF-κB p65 protein were determined in renal tissue of rats. RESULTS Compared with the normal group, the model group suffered from the dilatation of renal tubules, structural damage to glomeruli, accompanied by inflammatory infiltration and fibrosis; the contents of uric acid, Cr, BUN and XOD, the protein expressions of MCP-1,TNF-α and IL-6 and the phosphorylation levels of p38 MAPK and NF-κB p65 protein were all increased significantly (P<0.05 or P<0.01). Compared with the model group, the pathological symptoms of renal tissue in rats had been improved to varying degrees in different dose groups of PAP; the contents of uric acid, Cr, BUN and XOD, protein expressions of MCP-1, TNF-α and IL-6, the phosphorylation levels of p38 MAPK and NF-κB p65 protein in PAP high-dose and PAP medium-dose groups were all decreased significantly (P<0.05 or P<0.01). CONCLUSIONS PAP exhibits an anti-GN effect, the mechanism of which may be associated with inhibiting the p38 MAPK/NF-κB/TNF-α signaling pathway.
6.Trends in areas of Oncomelania hupensis snail habitats in Anhui Province from 2004 to 2023 based on the Mann-Kendall test
Fenghua GAO ; Shiqing ZHANG ; Songjun DING ; Xiaojuan XU ; Tianping WANG ; Bo DAI
Chinese Journal of Schistosomiasis Control 2024;36(5):460-465
Objective To investigate the trends in areas of Oncomelania hupensis snail habitats in Anhui Province from 2004 to 2023, so as to provide insights into facilitating the progress of schistosomiasis elimination in the province. Methods Areas of O. hupensis snail habitats in Anhui Province from 2004 to 2023 were collected, and the trends in and mutations of O. hupensis snail habitats in Anhui Province, and the trends in O. hupensis snail habitats in different types of schistosomiasis-endemic foci and in different cities of Anhui Province from 2004 to 2023 were analyzed using Mann-Kendall test. Results The areas of O. hupensis snail habitats reduced by 12.62% in Anhui Province in 2023 relative to in 2004. Mann-Kendall trend test showed that the areas of O. hupensis snail habitats appeared a tendency towards a decline in Anhui Province from 2004 to 2023 (Z = −3.926, P < 0.001, β = −215.66), with an annual average reduction of 215.66 hm2. Mann-Kendall mutation test showed that the areas of O. hupensis snail habitats appeared a tendency towards a decline in Anhui Province since 2015 (|Z| > U0.05, P < 0.05). The areas of O. hupensis snail habitats appeared a tendency towards a decline in marshland and lake regions (Z = −4.899, P < 0.001, β = −260.13), and a tendency towards a rise in hilly regions of Anhui Province from 2004 to 2023 (Z = 2.758, P < 0.01, β = 44.55). Among 9 cities endemic for schistosomiasis in Anhui Province, the areas of O. hupensis snail habitats appeared a tendency towards a decline in 5 cities of Ma’anshan, Tongling, Anqing, Huangshan and Chizhou (Z = −4.056 to −2.109, all P values < 0.05, β = −92.12 to −0.45). The average constituent ratios of areas of O. hupensis snail habitats were 88.19% and 11.81% in marshland and lake regions and hilly regions of Anhui Province from 2004 to 2023, respectively, and the ratio of areas of O. hupensis snail habitats in marshland and lake regions to areas of snail habitats in hilly regions reduced from 9.23∶1 in 2004 to 5.74∶1 in 2023 in Anhui Province, with the ratio appearing a tendency towards a decline (Z = −4.477, P < 0.001, β = −0.16). Conclusions The areas of O. hupensis snail habitats appeared a tendency towards a decline in Anhui Province from 2004 to 2023, with a reduction in areas of O. hupensis snail habitats in marshland and lake regions and a rise in hilly regions. Targeted O. hupensis snail control interventions are required tailored to real snail status.
7.Cost-effectiveness analysis of chemical treatment with drones for Oncomelania hupensis control in marshland and lake areas
Yong CHEN ; Xiaojuan XU ; Daolong WEN ; Bo DAI ; Lan GAO ; Rong ZHANG ; Qingqing HUANG ; Linlin LI ; Fan ZHA ; Liang FANG ; Ping ZHANG ; Shiqing ZHANG ; Chunli CAO
Chinese Journal of Schistosomiasis Control 2024;36(5):502-506
Objective To evaluate the molluscicidal effect and cost of spraying molluscicides with drones against Oncomelania hupensis snails in marshland and lake areas, so as to provide new insights into field snail control in China. Methods A marshland and lake setting measuring approximately 12 000 m2 was selected in Wanzhi District, Wuhu City on June 2023 as the test field, and assigned to four groups, of 3 000 m2 in each group. Environmental cleaning was not conducted in groups A or B, which were given 5% niclosamide ethanolamine salt granules sprayed with knapsack-type sprayers and drones at a dose of 40 g/m2, and environmental cleaning was conducted in groups C and D, which were given 5% niclosamide ethanolamine salt granules sprayed with drones and knapsack-type sprayers at a dose of 40 g/m2, respectively. O. hupensis snails were surveyed before chemical treatment and 1, 3, 5, 7, 14 days post-treatment. The uniformity of chemicals was determined on the day of treatment, and the snail mortality, corrected snail mortality and density of living snails were calculated and compared among groups. The cost of molluscicides, labor fees of environmental cleaning and chemical treatment and cost of equipment were calculated, and the cost for a 1% reduction in the mean density of living snails was calculated 14 days post-treatment. Results The mean densities of living snails and mortality rates of snails were 1.82 to 2.85 snails/0.1 m2 and 1.41% to 2.94% in groups A, B, C and D before chemical treatment, and the mortality and corrected mortality of snails were 55.75%, 49.32%, 85.94% and 87.50%, and 55.00%, 48.47%, 85.70% and 87.29% in groups A, B, C and D 14 days post-treatment. There was a significant difference in the mortality of snails among the four groups 14 days post-treatment (χ2 = 38.735, P < 0.005), and there was a higher snail mortality in Group D than in Group A (χ2 = 16.876, P < 0.005), and higher in Group C than in Group B (χ2 = 20.508, P < 0.005). The density of living snails reduced by 55.00%, 43.94%, 90.43% and 87.14% 14 days post-treatment relative to pre-treatment in groups A, B, C and D, respectively. The test for uniformity of chemicals showed that the mean dose of molluscicides were 57.34, 55.21, 40.19 g/m2 and 32.37 g/m2 in groups A, B, C and D, respectively, and the minimal standard deviation (7.07) and coefficient of variation (0.18) of mean doses were seen in Group C. The costs for chemical treatment were 0.33 Yuan in groups A and B and 1.53 Yuan in groups C and D, respectively. The costs for a 1% reduction in the mean density of living snails were 17.82, 22.47, 50.73 Yuan and 52.56 Yuan in groups A, B, C, and D 14 days post-treatment, respectively. Conclusions The molluscicidal effect and cost of spraying 5% niclosamide ethanolamine salt granules with drones are comparable to manual spraying, and chemical treatment with drones are high in uniformity of molluscicides, time- and labor-saving, and feasible for applications in complex environments, which deserves widespread applications in the field of snail control.
8.Epidemic trends and spatiotemporal distribution characteristics of condyloma acuminatum in China from 2018 to 2023
Shiqing LIANG ; Zewei CHEN ; Xiaoli YUE ; Jing LI ; Jiahui ZHANG ; Xiangdong GONG
Chinese Journal of Epidemiology 2024;45(8):1073-1078
Objective:Analyzing the epidemic trends and spatio-temporal distribution characteristics of condyloma acuminatum in China from 2018 to 2023.Methods:Data of condyloma acuminatum cases and incidence rate including 31 provinces (autonomous regions and municipalities) from 2018 to 2023 were collected through the National Notifiable Infectious Disease Reporting System of China Information System for Disease Control and Prevention. Incidence trend analysis was conducted using Joinpoint 4.9.1 software, and spatial autocorrelation analysis using ArcGIS 10.5 software. Spatio-temporal scanning analysis was carried out with SaTScan 10.1.2 software.Results:The incidence rate of condyloma acuminatum declined from 7.26 per 100 000 in 2018 to 7.19 per 100 000 in 2023. The average annual percent change was -0.26%, which was no statistically significant downward trend ( t=-0.26, P=0.806). A significant positive global spatial autocorrelation was observed in the county-level incidence rate across the country, with the global Moran's I ranging from 0.55 to 0.60 (all P<0.001); the Getis-Ord General test statistic Z( G) was all >1.96, indicating a high-value clustering pattern in the reported incidence rate of condyloma acuminatum. The local spatial autocorrelation analysis detected 256, 244, 246, 284, 308, and 315 hotspots each year, which were mainly located in the provinces of Zhejiang, Fujian, Guangdong, Guizhou, Yunnan and Chongqing. Spatio-temporal scanning analyses identified 76 statistically significant spatiotemporal clusters covering 25 provinces (autonomous regions and municipalities). Conclusions:From 2018 to 2023, the reported incidence rate of condyloma acuminata in China exhibited a mild decline. The distribution of hotspot areas and spatiotemporal clusters was largely consistent, primarily in the southeastern coastal and southwestern regions.
9.A Diagnostic Study of Traditional Chinese and Western Medicine Based on the New England Journal Tongue Diagram
Xiaohui SONG ; Jiyao YIN ; Zichen LIU ; Guoquan LIU ; Zhuang GUO ; Shiqing QIU ; Jiajie XU ; Hongguan JIAO ; Weijun ZHANG ; Junwen WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(5):1354-1360
Objective Taking typical cases of Western medicine as an example,this paper explores the connection between Chinese and Western medicine on the understanding of tongue elephants.Methods After collecting the literature with tongue diagram attached to the clinical imaging column published in NEJM magazine,extracting the symptoms,signs and Western medicine disease information recorded in the literature,the tongue diagram was diagnosed from three aspects:tongue quality,tongue moss and sublingual meridians,and whether the symptoms and signs of tongue correspond to a certain diagnosis result,and the results were analyzed.Results A total of 48 articles were included,including 6 literature on abnormal tongue dynamics,which could correspond to abnormal tongue morphology in traditional Chinese medicine.Thirty-four cases of abnormal tongue shape were found.Among them,12 cases could be diagnosed with corresponding TCM tongue diagnosis,including 7 cases of abnormal tongue shape and 5 cases of abnormal coating.The remaining 22 cases were secondary changes in tongue structure.There were 8 articles on abnormal tongue color,including 1 abnormal tongue color,1 abnormal sublingual chord,and 6 abnormal lichen color.Conclusion Starting from the form and function,explore the connection between Chinese medicine and Western medicine in their understanding of tongue diagnosis,and promote the objectification and standardization of Chinese medicine tongue diagnosis.
10.Establishment and evaluation of non-obese polycystic ovary syndrome rat model
Shiqing QIAO ; Ting WANG ; Yonghuang YAN ; Jiusi YANG ; Yuling YU ; Yanmin WANG ; Yateng SUN ; Yujie WU ; Peixuan ZHU ; Min LI ; Cong CHEN ; Zeqi SU ; Cai ZHANG
Acta Laboratorium Animalis Scientia Sinica 2024;32(8):965-975
Objective To establish a stable rat model of non-obese polycystic ovary syndrome(PCOS)with clinical characteristics.Methods Dehydroepiandrosterone(DHEA)was used to establish a PCOS rat model by subcutaneous injection.Three-week-old female SD rats were divided into a normal group,6 mg/kg DHEA model group,and 60 mg/kg DHEA model group.The model groups were subcutaneously injected with the corresponding dose of DHEA daily,while the normal group was subcutaneously injected with glycerol daily for 21 consecutive days.The model was evaluated with ovarian histopathology as the gold standard to determine the optimal dosage of DHEA to induce a PCOS rat model.On this basis,the optimal DHEA modeling dose was selected,and stop and continue modeling groups were set up to observe the model for 28 days and evaluate its maintenance.The stop modeling group was no longer given DHEA,and the continued modeling group was subcutaneously injected with 60 mg/kg DHEA every 48 h.The evaluation indicators included body mass,estrous cycle,fasting blood glucose,serum insulin,histopathologic morphology of the ovaries,and serum sex hormone levels.Results(1)Compared with the normal group,the 6 mg/kg and 60 mg/kg DHEA model groups showed no significant difference in body mass,and their estrous cycles were irregular.There were more cystically dilated large follicles in the ovaries;fewer mature follicles;reduced layers of granulosa cells,which were arranged in a sparse and disorganized manner;and fewer lutea in the 6 mg/kg and 60 mg/kg DHEA model groups than the normal group.Furthermore,serum T and E2 levels were significantly higher in the 60 mg/kg DHEA model group(P<0.05)than the normal group.(2)The stop modeling group(A2 group)resumed regular estrous cycles after 2 weeks,various growth follicles and corpora lutea were observed in the ovarian tissues,the number of cystic follicles was reduced,the number of granulosa cell layers increased,mature follicles were visible,oocyte morphology was locally intact,and the levels of E2 and AMH were reduced compared with the normal group(A1 group)(P<0.05).(3)The continue model group(B2 group)was in the late stage of estrous cycle for a long period,and there were more large follicles with cystic dilatation,fewer mature follicles,fewer layers of granulosa cells with a sparse and disordered arrangement,and significantly fewer corpus lutea in the ovaries compared with the normal group(B1 group).The levels of serum LH,LH/FSH,and T were elevated(P<0.05).Conclusions Subcutaneous injection of 60 mg/kg DHEA for 21 consecutive days can be used to successfully construct a non-obese PCOS rat model that possesses clinical characteristics.Subcutaneous injection of 60 mg/kg DHEA every 48 hours maintains the stability of the model.

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