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Nucleic acid-based molecular diagnostic methods are considered the gold standard for detecting infectious pathogens.However,when applied to portable or on-site rapid diagnostics,they still face various limitations and challenges,such as poor specificity,cumbersome operation,and portability difficulties.The CRISPR(Clustered regularly interspaced short palindromic repeats)/CRISPR-associated protein(Cas)-fluorescence detection method holds the potential to significantly enhance the specificity and signal-to-noise ratio of nucleic acid detection.In this study,we developed a portable grayscale reader detection system based on loop-mediated isothermal amplification(LAMP)-CRISPR/Cas.On one hand,in the presence of CRISPR RNA(crRNA),the CRISPR/Cas12a system was employed to achieve precise fluorescent detection of self-designed LAMP amplification reactions for influenza A and influenza B viruses.This further validated the high selectivity and versatility of the CRISPR/Cas system.On the other hand,the accompanying independently developed portable grayscale reader allowed for low-cost collection of fluorescence signals and high-reliability visual interpretation.At the end of the detection process,it directly provided positive or negative results.Practical sample analyses using this detection system have verified its reliability and utility,demonstrating that this system can achieve highly sensitive and highly specific portable analysis of influenza viruses.
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Osteoarthritis (OA) is the most common chronic disabling joint disease, and currently there is no effective treatment for the cause. Necroptosis plays a key role in many diseases, and receptor-interacting protein kinase 3 (RIP3) is a key regulator during necroptosis process. Studies have shown that the expression level of RIP3 was significantly upregulated in human and mouse OA degenerative cartilage tissues, suggesting the occurrence of necroptosis. However, the specific pathophysiological role of RIP3 in cartilage is still unclear. This study intends to sequence and analyze the transcriptome of chondrocytes before and after RIP3 overexpression, and explore the specific functional mechanism of RIP3 in OA pathogenesis. RNA sequencing results showed that overexpression of RIP3 induced upregulation of 244 genes and downregulation of 277 genes in chondrocytes. Sixteen candidate target genes were screened out by constructing gene co-expression network for further verification at mRNA level, and the results suggested that RIP3 had the most significant inductive effect on the expression of phosphoinositide-3kinase, regulatory subunit 5 (Pik3r5), integrin subunit beta 3 (Itgb3) and MYB proto-oncogene like 2 (Mybl2). Results from CCK-8 and lactate dehydrogenase activity analysis showed that silencing the expression of Itgb3 by siRNA significantly rescued chondrocyte viability decline and necroptosis induced by RIP3, and it also inhibited the upregulating effect of RIP3 on the expression of catabolism-related genes Mmp1, Mmp13 and Il6, as well as the downregulating effect of RIP3 on the expression of anabolism-related genes Acan, Col2a1 and Sox9. This study has demonstrated that RIP3 promotes chondrocyte necrosis and cartilage matrix metabolism disorders by upregulating the expression of Itgb3 in chondrocytes, and ultimately leads to cartilage degeneration. These findings provided potential novel targets for the clinical treatment of OA, and further clarified the pathophysiological significance of necroptosis.
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OBJECTIVE@#To make a retrospective analysis of the situation and process of treating skiers' injuries in the medical station of the Wanlong ski resort in Chongli and the nearest treatment hospital, and to provide a basis for the establishment and optimization of the treatment process between the medical station of the Winter Olympics ski resort and the nearest treatment hospital, and to gain experience for medical security of mass skiing.@*METHODS@#The data of all ski injuries in Chongli District were collected from the medical station of the Wanlong ski resort during the 2018-2019 snow season (November 2018 to April 2019) and the nearest treatment hospital during two periods (March 2019, and November 2019 to January 2020). The differences of injury causes, injury types, injury sites, and treatment effects of the injured skiers were analyzed.@*RESULTS@#A total of 755 cases of ski injuries were recorded in the medical station of Wanlong ski resort, the estimated incidence of injury was 2.02‰ per day. The nearest treatment hospital treated a total of 838 injured skiers from different ski resorts in Chongli District in the two periods. In the records of the ski resort medical station, the main causes of injury were technical defects and turnovers (53.6%). Knee joint injury rate was the highest (18.7%), followed by head and neck (12.9%) and lower limb (11.9%). The number of injuries on intermediate roads was the highest (40.0%), the greatest number of injuries (81.2%) occurred when the age of skiing was less than 5 years. In the records of the nearest treatment hospital, the injury types were fracture or fissure fracture, contusion and trauma, and muscle and soft tissue injury, accounting for 30.5%, 27.4%, and 21.2% respectively. 9.6% of the injured took the snow field ambulance to the hospital, and 50% of them suffered from fractures or fissure fractures.@*CONCLUSION@#The injury rate of skiing in the 2018-2019 snow season of the Wanlong ski resort in Chongli was higher than that reported by foreign literature. Severe trauma (including severe fractures and concussions) could occur and patients needed to be transferred to the nearest hospital for treatment. The ski resort medical station and the nearest treatment hospital should be strengthened with adequate medical staff and equipment, and promote cooperation in the timely referral of seriously injured patients, the organization and construction of ski patrols and the medical security of large-scale competitions, thus playing an important role in forming a grassroots network of medical security and treatment system for skiing.
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Pré-Escolar , Humanos , Traumatismos em Atletas/terapia , Hospitais , Estudos Retrospectivos , Estações do Ano , EsquiRESUMO
OBJECTIVE@#To compare and analyze the difference between the injuries of recreational skiers in public ski resorts and those of skiing athletes in official competitions, and to explore the suggestions of medical insurance for these two types of skiers.@*METHODS@#The injury data of recreational skiers in Chongli District, Zhangjiakou City, Hebei Province during 2018-2019 and 2019-2020 snow seasons, and the injury data of skiers in two official international skiing competitions during 2019-2020 snow season and domestic test events in Chongli District of 2021 Winter Olympic Games were analyzed retrospectively, and the similarities and differences were compared.@*RESULTS@#A total of 1 187 injuries occurred to recreational skiers in the two public ski resorts during the 2018-2019 snow season, with an injury rate of 0.3%.There were 1 277 injury sites in total, and the most frequent injury sites were head and neck (230 cases, 18.0%), followed by knee joint (204 cases, 16.0%) and lower extremity (131 cases, 10.3%). Thirty-one skiers were injured in the two official international skiing competitions in the 2019-2020 and 2020-2021 snow season, and in the domestic test competitions in the 2021 Winter Olympic Games, and the injury rates were 11.5%, 17.2% and 12.0%, respectively. There were 37 injury sites in total, among which 11 (29.7%) were in the head and neck, followed by 6 (16.2%) in the knee joint and 5 (13.6%) in the chest, rib and abdomen.@*CONCLUSION@#In order to better guarantee the safety of skiers and timely provide corresponding medical help, safety facilities and technical guidance should be added to the snow resort for leisure skiing, and medical stations should be set up in the snow resort. As the formal ski racing for skiing athletes during the game has 30 to 80 times higher injury ratesthan recreational skiers, and compared with the recreational skiing, head and neck injury rate is higher, and the damage is much heavier, more complete first aid facilities and experienced medical workers are, needed so the field should be equipped with the circuit inside the quantities, track fixed outside the clinic, surrounding referral hospitals set up trauma centers to provide athletes with more timely medical care.
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Humanos , Traumatismos em Atletas/epidemiologia , Extremidade Inferior , Estudos Retrospectivos , Esqui , Centros de TraumatologiaRESUMO
OBJECTIVE@#To assess the mid-to-long term clinical outcomes after anterior cruciate ligament (ACL) revision surgery and to analyze their predictors.@*METHODS@#The medical records of 235 patients undergoing ACL revision surgery between Jan. 2001 and Dec. 2015 at Department of Sports Medicine, Peking University Third Hospital were reviewed. Data were collected including demographic information, information related to revision surgery (time and cause of graft failure, date of revision surgery, surgical technique, combined injuries and management, etc.), as well as information related to primary ACL reconstruction (time, cause and mechanism of first-time ACL rupture, date of primary ACL reconstruction, surgical technique, combined injuries and management, etc.). Patients were followed up at least 2 years after revision surgery for clinical outcomes [Tegner score, Lysholm score, and International Knee Documentation Committee (IKDC) subjective knee score]. Post-revision surgeries on the involved knee and the contralateral knee joint were also documented. Multivariate regression model was used to analyze the predictors of clinical outcomes after ACL revision surgery.@*RESULTS@#A total of 166 (70.63%) patients were followed up at a mean of (4.44±2.40) years (2.03-14.63 years). Clinical outcomes improved significantly at the last follow-up from pre-operative level, with the Lysholm, Tegner, and IKDC scores improving from 70.51±21.25, 3.39±1.77, 63.78±15.04 to 88.64±14.36, 4.67±1.739, 80.23±13.31 (P < 0.05), respectively. Three (1.81%) patients experienced infection while 39 (23.49%) patients underwent surgery after revision surgery during the follow-up. Compared with that those occurred during sports, graft failure that occurred during daily activities or due to surgical technical errors that led to poorer clinical outcomes, with the Lysholm, Tegner, and IKDC scores of 9.90 (95%CI: 1.49-18.31), 1.41 (95%CI: 0.10-2.72), 10.35 (95%CI: 0.17-20.54), and 8.53 (95%CI: 1.31-15.75), 1.28 (95%CI: 0.14-2.43), 9.39 (95%CI: 1.03-17.74) lower, respectively. Compared with antero-medial portal, transtibial technique for placement of the femoral bone tunnel showed poorer Lysholm scores of 11.18 (95%CI: 4.73-17.63, P=0.001). Concurrent repair of medial meniscus yielded higher IKDC scores of 11.06 (95%CI: 1.21-20.92, P=0.029) than those with intact medical meniscus. Other factors showed no significant effect.@*CONCLUSION@#ACL revision surgery is able to restore knee stability and improve knee function. Graft failure caused by sports, concurrent repair of medical meniscus and antero-medial portal technique predicts better outcomes after revision surgery.
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Humanos , Ligamento Cruzado Anterior/cirurgia , Lesões do Ligamento Cruzado Anterior/cirurgia , Reconstrução do Ligamento Cruzado Anterior , Articulação do Joelho/cirurgia , Reoperação , Resultado do TratamentoRESUMO
OBJECTIVE@#To analyze the preoperative pulmonary function in rotator cuff injury patients and the possible influencing factors.@*METHODS@#All the rotator cuff surgery patients who underwent pre-operative pulmonary function examination in Peking University Third Hospital from Jan. 2020 to Jun. 2020 were retrospectively reviewed. Their perioperative medical records and main parameters of pulmonary function were collected from database management system, and their gender, age, body mass index (BMI), smoking history, time from injury, visual analogue scale (VAS) and other factors impacting on preoperative pulmonary function were studied.@*RESULTS@#Twenty-nine patients with rotator cuff injury were included, among whom 1 patient was reported to have restrictive ventilation dysfunction and 2 patients to have obstructive ventilation dysfunction. All the three patients denied the history of respiratory diseases, and had no respiratory symptoms. In all enrolled patients, the mean forced expiratory volume in one second (FEV1)/ forced vital capacity (FVC) was 79.2%±5.9%, and the mean VAS pain score was 3.66±1.26. In addition, the dynamic pulmonary functions (FVC, FEV1) were reduced in more than half of the elderly, and the total lung capacity (TLC) was lower than the estimated value in 2/3 of the elderly. There were significant differences in three main indexes of pulmonary functions between genders, and the percentage of the estimated TLC between normal BMI group (18 kg/m2 < BMI < 24 kg/m2) and overweight/obesity group (BMI≥24 kg/m2) was significantly different. Based on the injury time longer than 1 year or not, the results indicated that FVC and TLC were significantly different between the two groups.@*CONCLUSION@#In addition to gender and age, time from injury and severity of pain, as well as overweight/obesity, may influence pulmonary function outcomes in the elderly rotator cuff patients. Targeted intervention can be carried out on these factors before surgery. Preoperative lung function test can be used as one of the basic evaluation indexes for respiratory training and rehabilitation of patients.
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Idoso , Feminino , Humanos , Masculino , Pulmão , Testes de Função Respiratória , Estudos Retrospectivos , Manguito Rotador , Lesões do Manguito Rotador/cirurgiaRESUMO
OBJECTIVE@#To make a retrospective analysis of the injuries of skiing population in a large ski resort in Chongli, China and provide a basis for predicting the rapidly increasing medical needs for ski injuries in the context of the 2022 Winter Olympic Games.@*METHODS@#The basic data of all injured skiers who were treated in a medical station of a large ski resort in Chongli during the snow season from November 2017 to March 2018 and from November 2018 to March 2019 were collected. The number of skiers, the number of injuries, the causes of injuries, the types of injuries and the locations of injuries were compared.@*RESULTS@#A total of 753 skiers were injured in two snow seasons, and the estimated average incidence of injury was 4.53 and 4.46 per 1 000 skier days at the resort respectively. The average daily injury rate per 1 000 skiers in November of the two snow seasons was relatively low, with 2.20 and 1.38 cases respectively. The difference of injury rate in different months might have little to do with snowfall and more to do with passenger flow. In both the snow seasons, men accounted for more injuries than women, and injured skiers aged between 21 and 30 accounted for the largest proportion, reaching 36.8%. The main causes of injuries were falls (76.6%). The highest rate of injury was in the head and neck (17.9%), followed by the knee (17.4%) and wrist and fingers (13.3%). The most common types of injuries were contusion and trauma (29.5%) and joint and/or ligament injuries (22.2%). Children (2-12 years old) accounted for 12.7% of all the injured skiers. The rate of moderate to severe injuries (including fractures, concussions, etc.) was 34.8% among the injured patients over 50 years of age.@*CONCLUSION@#The snow resort should focus on injuries to children and elderly skiers and carry out targeted guidance and rescue work. In order to better ensure the medical safety of skiers, the ski resort medical station and nearby treatment hospitals should be equipped with a corresponding number of medical personnel and equipment, and the ski resort should further improve its safety management and rescue system.
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Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Adulto Jovem , Traumatismos em Atletas/epidemiologia , China/epidemiologia , Incidência , Estudos Retrospectivos , EsquiRESUMO
OBJECTIVE@#To investigate the effects of quercetin on the apoptosis of platelets and to analyze the intrinsic mechanism.@*METHODS@#Firstly, the effects of quecetin on the apoptosis of platelets was detected by flow cytometry. Secondly, Western blot was used to detect the expression of apoptosis-related proteins in the platelets treated with quercetin for 2 and 4 day.@*RESULTS@#By flow cytometry, it was found that the apoptosis of platelets in the quercetin-treated group (2, 4 and 8 μmol/L) was inhibited, the apoptosis rate of platelets in 2, 4 and 8 μmol/L quercetin group was 3.12%±0.32%, 2.89%±0.15% and 2.31%±0.28%, respectively, which were signigicantly lover than that in control group (P<0.01). With the increase of quecetin concentration, the proportion ratio of platelets significantly decreased in a concentration-dependent manner(r=-0.9985). Similar results were observed on the 4th day. Western blot showed that the treatment with quercetin (2, 4 and 8 μmol/L) promoted the expression of anti-apoptotic protein BCL-2, inhibited the expression of pro-apoptotic protein BAX, resulting in a significant increase in the ratio of BCL-2/BAX (P<0.01), thereby inhibiting the apoptosis of platelets. Similar results were observed on the 4th day.@*CONCLUSION@#Quercetin can inhibit platelet apoptosis by increasing the ratio of apoptosis-related protein BCL-2/BAX in a concentration-dependent manner.
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Apoptose , Proteínas Reguladoras de Apoptose , Plaquetas , QuercetinaRESUMO
<p><b>BACKGROUND</b>Primary biliary cirrhosis (PBC) is a chronic and slowly progressive cholestatic liver disease characterized by destruction of the interlobular bile ducts and a striking female predominance. The aim of this study was to identify associations between estrogen receptor (ESR) gene polymorphisms with the risk of developing PBC and abnormal serum liver tests in a Chinese population.</p><p><b>METHODS</b>Thirty-six patients with PBC (case group) and 35 healthy individuals (control group) from the First Hospital of Jilin University were studied. Whole genomic DNA was extracted from all the participants. Three single-nucleotide polymorphisms (rs2234693, rs2228480, and rs3798577) from ESR1 and two (rs1256030 and rs1048315) from ESR2 were analyzed by a pyrosequencing method. Demographic data and liver biochemical data were collected.</p><p><b>RESULTS</b>Subjects with the T allele at ESR2 rs1256030 had 1.5 times higher risk of developing PBC than those with the C allele (odds ratio [OR] = 2.1277, 95% confidence interval [CI] = 1.1872-4.5517). Haplotypes TGC of ESR1 rs2234693, rs2228480, and rs3798577 were risk factors for having PBC. The C allele at ESR1 rs2234693 was associated with abnormal alkaline phosphatase (OR = 5.2469, 95% CI = 1.3704-20.0895) and gamma-glutamyl transferase (OR = 3.4286, 95% CI = 1.0083-13.6578) levels in PBC patients.</p><p><b>CONCLUSIONS</b>ESR2 rs1256030 T allele may be a significant risk factor for the development of PBC. Screening for patients with gene polymorphisms may help to make early diagnoses in patients with PBC.</p>
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Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Povo Asiático , Estudos de Casos e Controles , Receptor alfa de Estrogênio , Genética , Receptor beta de Estrogênio , Genética , Frequência do Gene , Genética , Predisposição Genética para Doença , Genética , Haplótipos , Genética , Cirrose Hepática Biliar , Genética , Polimorfismo de Nucleotídeo Único , Genética , Receptores de Estrogênio , GenéticaRESUMO
<p><b>BACKGROUND</b>Chronic liver diseases are a major burden in China. Alanine aminotransferase (ALT) can be used as an indicator of hepatocyte damage. In this study, we determined the prevalence and etiologies of elevated ALT in an adult population in Jilin, China.</p><p><b>METHODS</b>A total of 4072 individuals aged between 18 and 79 years were first interviewed, and then underwent ultrasonography and blood tests.</p><p><b>RESULTS</b>The prevalence of elevated ALT was 17.53%. The most noticeable risk factor for ALT elevation was non-alcoholic fatty liver disease (NAFLD) (accounting for 10.79%), metabolic syndrome (16.25%), or both (20.31%). The development of NAFLD occurred mostly in female peasants and small businessmen with increased income, age, fasting plasma glucose, body mass index, triglyceridemia, and low-density lipoprotein and decreased education level, high-density lipoprotein. Elevated ALT frequently occurred in low education level, male peasants and small businessmen with increased income, body mass index and triglyceride who had NAFLD and/or metabolic syndrome. However, elevated ALT with infection of hepatitis B or C virus was not associated with metabolic disorders, but rather with gender, occupation and increased age.</p><p><b>CONCLUSION</b>The results from the current study demonstrate that elevated ALT is fairly high in the Northeast population (17.53%) and that the cause of its elevation is mostly due to NAFLD and metabolic syndrome.</p>
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Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Fatores Etários , Alanina Transaminase , Sangue , Fígado Gorduroso , Sangue , Epidemiologia , Hepatite B , Sangue , Epidemiologia , Hepatite C , Sangue , Epidemiologia , Síndrome Metabólica , Sangue , Hepatopatia Gordurosa não Alcoólica , Fatores SexuaisRESUMO
<p><b>OBJECTIVE</b>To investigate the inhibitory effect of dehydroepaimdrosterone (DHEA) on the growth of transplanted Morris hepatomas (7288CTC) in vivo in rats.</p><p><b>METHODS</b>21 Buffalo rats were randomly devided into 4 groups, including one blank control group (n = 5), one group for tumor-bearing control (n = 6), and 2 experimental groups with DHEA (n = 6) or DHEA-s (n = 4). DHEA or DHEA-s was fed to the rats for 4 weeks immediately after Morris hepatomas (7288CTC) was implanted in both flanks. Phenotypes of the spleen lymphocytes were examined by flow cytometry, Akt and PTEN expression in tumor cells was detected by Western blot and immunohistochemistry.</p><p><b>RESULTS</b>Tumor weights of DHEA treated group were less than those of the control (P less than 0.05), the inhibitory rate was 43%. The results of Western blot and immunohistochemistry showed that in DHEA tumor group,the expression of phosphorilated Akt protein was decreased, the expression of PTEN was enhanced, the percentage of CD3 positive cells and the ratio of CD4/CD8 were increased (P less than 0.05).</p><p><b>CONCLUSION</b>DHEA can inhibit tumor growth, possibly via the inhibition of the Akt signaling pathway as well as modulating the immune function.</p>
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Animais , Ratos , Antineoplásicos , Farmacologia , Linfócitos T CD4-Positivos , Alergia e Imunologia , Linfócitos T CD8-Positivos , Alergia e Imunologia , Desidroepiandrosterona , Farmacologia , Sulfato de Desidroepiandrosterona , Farmacologia , Citometria de Fluxo , Imuno-Histoquímica , Neoplasias Hepáticas Experimentais , Alergia e Imunologia , Metabolismo , Patologia , Transplante de Neoplasias , PTEN Fosfo-Hidrolase , Metabolismo , Proteínas Proto-Oncogênicas c-akt , Metabolismo , Distribuição Aleatória , Ratos Endogâmicos BUF , Transdução de SinaisRESUMO
<p><b>OBJECTIVE</b>To evaluate the anti-proliferation effects of dehydroepiandrosterone (DHEAéand DHEA sulfate (DHEAs) on tumor cells.</p><p><b>METHODS</b>Human hepatoblastoma cells (HepG2) and colon adenocarcinoma cells (HT-29) were treated with DHEA and DHEAs of various concentrations. The cells were incubated for 8, 24, 48, and 72 hours, and the proliferation, apoptosis, cell cycle and the expression of phosphorylated Akt (Thr308 and Ser473) were analyzed using MTT assay, flow cytometry, and Western blotting at different time points. The influences of an inhibitor (LY294002) and an activator (hepatic growth factor; HGF) of PI3K on the effectiveness of DHEA were determined in HepG2 cells.</p><p><b>RESULTS</b>By increasing the concentrations of DHEA (1, 10, 50, 100, 200 micromol/L), the percentages of HepG2 and HT-29 survival cells treated with DHEA at 24 h were 92.7%+/-0.9%, 84.7%+/-1.2%, 62.4%+/-0.8%, 49.5%+/-0.8%, 50.7%+/-0.3% and 92.5%+/-0.4%, 89.5%+/-0.7%, 80.5%+/-1.1%, 67.5%+/-1.5%, 70.6%+/-0.6%, respectively. Proliferations of HepG2 and HT-29 cells were significantly inhibited after 24 hours of being incubated with 100 micromol/L DHEA treatment; the inhibition effect was stronger on HepG2 cells than on HT-29 cells. The effect of DHEAs on both cell lines on cell proliferation was weaker than that of the DHEA. In the cell cycle assay, DHEA treatment induced cell arrest in G0/G1 phase in both cell lines. Apoptosis of HepG2 cells was significantly triggered (18.6%+/-2.2%) by 100 micromol/L DHEA treatment for 24 hours, but not by DHEAs. In addition, 100 and 200 micromol/L DHEA treatments for 24 hours markedly inhibited phosphorylations of Akt (Thr308 and Ser473) in HepG2 cells, and these effects were enhanced by exposing them to LY294002 and stopped by exposing them to HGF. The anti-proliferative effects of DHEA on tumor cell lines were much stronger than those of DHEAs, and they were even stronger in HepG2 cells than in HT-29 cells.</p><p><b>CONCLUSION</b>Our results suggest that the induction of apoptosis through the inhibition of Akt signaling pathway is one of the anti-proliferative mechanisms of DHEA in certain tumors, but DHEA also promotes cell-cycle arrest.</p>