1.Aspirin alleviates OGD/R-induced injury in mouse hippocampal neurons by regulating ferroptosis
Yujiao HU ; Shan CONG ; Lei ZHAO ; Chunxue DONG ; Dongmei WANG ; Nannan WANG ; Ying MAO
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(8):960-964
Objective To investigate the effect of aspirin on oxygen glucose deprivation/reoxygen-ation(OGD/R)-induced injury in mouse hippocampal neuron HT22 cells by regulating ferropto-sis.Methods HT22 cells were randomly divided into control group,model group,low-,medium-and high-dose groups(n=3).Cellular OGD/R injury model was established in the other 4 groups except the control group.Aspirin of 100,200 and 400 μg/ml was used to treat the cells from the above 3 treatment groups,respectively.Cell viability was detected by CCK-8 assay.The contents of TNF-α,IL-1β and IL-6 were detected by ELISA.The contents of SOD,catalase,glutathione,reac-tive oxygen species(ROS),lactate dehydrogenase(LDH),Fe2+and MDA were detected by the corresponding reagent kits.Western blot analysis was performed to determine the expression of solute carrier family 7 members 11(SLC7A11),glutathione peroxidase 4(GPX4)and Acyl-CoA synthase long chain member 4(ACSL4).Results The model group had significantly lower cell vi-ability than the control group(0.49±0.07 vs 1.00±0.12,P<0.01),but the viability of the low-,medium-and high-dose groups were higher than that of the model group(0.72±0.10 vs 0.49±0.07,P<0.05;0.87±0.10 vs 0.49±0.07,P<0.01;0.93±0.07 vs 0.49±0.07,P<0.01).Compared with the control group,the contents of TNF-α,IL-1β,IL-6,ROS,LDH,Fe2 and MDA and the protein expression of ACSL4 were significantly increased,while the contents of SOD,catalase,glutathione and protein levels of SLC7A11 and GPX4 were obviously decreased in the model group(P<0.01).Compared with model group,aspirin treatment reversed all above indicators no matter its doses(P<0.05,P<0.01).Conclusion Aspirin alleviates OGD/R-induced neuronal in-jury through regulating ferroptosis in mouse neuron HT22 cells in a dose-dependent manner.
2.Expert consensus on the management of auditory hallucinations in inpatients with mental illness
Yanhua QU ; Dongmei XU ; Jing SHAO ; Shan ZHANG ; Mengqian ZHANG ; Jianing GU ; Xiaolu YE ; Feifei LI ; Wei LUO ; Wanting LI ; Li WANG ; Fangzhu SHI ; Xiaoyu FENG ; Qian ZHOU ; Juan ZHAO
Chinese Journal of Practical Nursing 2024;40(14):1080-1090
Objective:To standardize the management of auditory hallucination symptoms in inpatients with mental illness and develop an expert consensus on the management of auditory hallucinations in hospitalized psychiatric patients.Methods:From March 2023 to July 2023, the Mental Health Committee of the Chinese Nursing Association focused on the key issues in the management of auditory hallucinations symptoms in inpatients with mental illness, based on clinical practice, using literature analysis combined with the work experience of mental health experts, and formed the first draft of the expert consensus on the management of auditory hallucinations in inpatients with mental illness (hereinafter referred to as the consensus). Through 3 rounds of expert consultation and 3 rounds of expert demonstration meeting, the draft was adjusted, revised, and improved.Results:37 experts were included in the Delphi expert consultation, 1 male and 36 females with 39-67(51.48 ± 6.61) years old. The positive coefficients of experts in 3 rounds of Delphi expert consultations were all 100%, and the degrees of expert authority were 0.924, 0.938 and 0.949, respectively. The average importance value of each item was higher than 4.00, the variation coefficient of each item was less than 0.25. The Kendall harmony coefficient of the experts were 0.179, 0.195 and 0.198, respectively (all P<0.05). There were 15, 12, 12 experts in the first, seeond, third rounds of expert demonstration meeting. Finally, a consensus was reached on the recommendation of 4 parts, included auditory hallucination assessment, management format, symptom management implementation, and precautions. Conclusions:The consensus covers all parts of the management of auditory hallucination symptoms in hospitalized patients with mental disorders, which is practical and scientific. It is helpful to guide mental health professionals to standardize the management of auditory hallucination symptoms, improve the quality of nursing and ensure the safety of patients.
3.Application of flat-sided culture tubes during prenatal diagnosis.
Tiansheng LIU ; Hongqian HUANG ; Jiangyu SU ; Wangshang QIN ; Dongmei FEI ; Luping OUYANG ; Minpan HUANG ; Jinwu YU ; Yaqin LEI ; Shan OU ; Weijia SUN ; Qingming QIU ; Qian ZHENG
Chinese Journal of Medical Genetics 2023;40(10):1306-1311
OBJECTIVE:
To assess the value of using flat-sided culture tubes for preparing chromosomes through chorionic villi (CV) and amniotic fluid (AF) cell cultures during prenatal diagnosis.
METHODS:
From February to March 2020, 157 CV samples and 147 AF samples subjected to prenatal diagnosis at the Maternal and Child Health Care Hospital of Guangxi Zhuang Autonomous Region were selected as the study subjects. For each sample, one flat-sided tube and one flask culture were set up by following the standard protocols. The methods were evaluated by comparing the cell growth, experimental process, quality of chromosome preparation and costs.
RESULTS:
The success rates for the culturing of CV and AF samples by the flat-sided culture tube method were 97.45% (153/157) and 97.96% (144/147), respectively. By contrast, the success rates for the conventional flask method were 98.72% (155/157) for CV and 98.64% (145/147) for AF samples. No significant difference was found between the two methods (P > 0.05). The average harvest time required by the flat-sided culture tube method was 8.45 days for CV and 9.43 days for AF cultures, whilst the average harvest time for conventional flask method was 9.05 days and 9.54 days, respectively. The flat-sided culture tube method for CV had required significantly shorter average harvest time than the conventional method (P < 0.001). No statistical significant difference was found in the average harvest time for AF by the two methods (P > 0.05). The conventional culturing method had required three containers with two sample transfers. By contrast, the flat-sided culture tube method was carried out in one tube without any sample transfer. The average total amount of medium used was 3.91 mL for each flat-sided culture tube and 6.26 mL for each conventional flask.
CONCLUSION
The flat-sided culture tube method can provide a simple, cost-effective and error-reducing procedure for the CV and AF samples culture during prenatal diagnosis.
Child
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Female
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Pregnancy
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Humans
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China
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Prenatal Diagnosis
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Chorionic Villi Sampling
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Amniotic Fluid
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Cell Proliferation
4.Effects of coronavirus disease 2019 vaccination on seizures in patients with epilepsy
Xiqin FANG ; Shan QIAO ; Ranran ZHANG ; Tingting YANG ; Zhihao WANG ; Qingxia KONG ; Meihua SUN ; Jianhong GENG ; Chunyan FANG ; Yanxiu CHEN ; Yanping SUN ; Dongmei ZHANG ; Lixing QU ; Wei SHANG ; Jianguo WANG ; Xuewu LIU
Chinese Medical Journal 2023;136(5):571-577
Background::Given that seizures may be triggered by vaccination, this study aimed to evaluate the risk and correlative factors of seizures in patients with epilepsy (PWE) after being vaccinated against coronavirus disease 2019 (COVID-19).Methods::This study retrospectively enrolled PWE who were vaccinated against COVID-19 in the epilepsy centers of 11 hospitals in China. We divided the PWE into two groups as follows: (1) patients who developed seizures within 14 days of vaccination were assigned to the SAV (with seizures after vaccination) group; (2) patients who were seizure-free within 14 days of vaccination were assigned to the SFAV (seizure-free after vaccination) group. To identify potential risk factors for seizure reccurence, the binary logistic regression analysis was performed. Besides, 67 PWE who had not been vaccinated were also included for elucidating the effects of vaccination on seizures recurrence, and binary logistic regression analysis was performed to determine whether vaccination would affect the recurrence rate of PWE who had drug reduction or withdrawal.Results::The study included a total of 407 patients; of which, 48 (11.8%) developed seizures within 14 days after vaccination (SAV group), whereas 359 (88.2%) remained seizure-free (SFAV group). The binary logistic regression analysis revealed that duration of seizure freedom ( P < 0.001) and withdrawal from anti-seizure medications (ASMs) or reduction in their dosage during the peri-vaccination period were significantly associated with the recurrence of seizures (odds ratio= 7.384, 95% confidence interval = 1.732–31.488, P = 0.007). In addition, 32 of 33 patients (97.0%) who were seizure-free for more than three months before vaccination and had a normal electroencephalogram before vaccination did not have any seizures within 14 days of vaccination. A total of 92 (22.6%) patients experienced non-epileptic adverse reactions after vaccination. Binary logistic regression analysis results showed that vaccine did not significantly affect the recurrence rate of PWE who had the behavior of ASMs dose reduction or withdrawal ( P = 0.143). Conclusions::PWE need protection from the COVID-19 vaccine. PWE who are seizure-free for >3 months before vaccination should be vaccinated. Whether the remaining PWE should be vaccinated depends on the local prevalence of COVID-19. Finally, PWE should avoid discontinuing ASMs or reducing their dosage during the peri-vaccination period.
5.Saponins in Treatment of Alzheimer's Disease and Their Mechanism of Action: A Review
Taoran LI ; Xiaoqing ZHOU ; Hongdong LIU ; Dongmei YAN ; Shan LIU ; Bin LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(14):216-225
Alzheimer's disease(AD) is an irreversible neurodegenerative disease with clinical manifestations such as memory impairment, aphasia, impaired visuospatial skills, executive function impairment, and personality changes. AD has brought a heavy burden to the family and society due to its unrevealed pathogenesis and the lack of therapeutic approaches. Saponins, a group of oligoglycosides whose aglycones are triterpenes or spirosteroids, are divided into triterpene saponins and steroidal saponins, which have a variety of biological activities. At present, there is no systematic review on the anti-AD effect of saponins. According to the literature published in recent years, the authors summarized the studies of saponins in improving AD based on animal experiments. The results indicated that saponins enhanced learning ability and improved cognitive impairment by inhibiting amyloid β-protein (Aβ) cascade activity, suppressing microtubule-associated protein (tau) hyperphosphorylation, inhibiting neuronal oxidative stress, inhibiting inflammatory factors, regulating apoptosis, inhibiting cholinergic neuronal degeneration, promoting mitochondrial autophagy, regulating intestinal flora, and enhancing energy metabolism, which in turn improved the pathological state of AD animal models. The therapeutic effects of different saponins on AD are different. The present study discussed the effect of different aglycones and sugar chains on the anti-AD activity based on saponins and anti-AD effect to provide new ideas and a theoretical basis for the development and utilization of saponins.
6.A comparison of clinical characteristics between acute fatty liver of pregnancy and hemolysis, elevated liver enzymes and low platelets syndrome
Dongmei DAI ; Shiyu TANG ; Wangbin XU ; Yuping WANG ; Leyun XIAOLI ; Xiao YANG ; Yancui ZHU ; Keji SHAN ; Linjun WAN ; Ming ZHU
Chinese Critical Care Medicine 2022;34(6):624-629
Objective:To compare and analyze the clinical characteristics between acute fatty liver of pregnancy (AFLP) and the hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome.Methods:This is a retrospective cohort study. The clinical data of 13 cases with AFLP and 34 cases with HELLP syndrome were collected from three tertiary referral centers in Yunnan (the First Affiliated Hospital of Kunming Medical University, the Second Affiliated Hospital of Kunming Medical University, and Yan'an Hospital of Kunming City) from January 2016 to December 2021. The patients were diagnosed to AFLP and HELLP syndrome according to the Swansea criteria and the Tennessee classification system. The general characteristics, clinical features, laboratory results within 24 hours after admission, complications, maternal and neonatal outcomes were compared to analysis the differences between the two groups.Results:① Maternal characteristics: compared with HELLP syndrome group, AFLP group had lower body mass index (BMI) and blood pressure at admission (both P < 0.01). ②Clinical features: the most common symptoms in AFLP patients were skin jaundice, abdominal pain, nausea and vomiting, edema. The main manifestations of patients with HELLP syndrome were albuminuria, hypertension, edema, headache. Some patients had multiple symptoms concurrently. ③ Laboratory results: compared with HELLP syndrome group, the levels of platelet count (PLT), total bilirubin (TBil), direct bilirubin (DBil), γ-glutamyl transferase (γ-GGT), alkaline phosphatase (ALP), total bile acid (TBA), serum creatinine (SCr) and international standardized ratio (INR) in AFLP group were significantly increased within 24 hours after admission [PLT (×10 9/L): 107.69±51.13 vs.76.71±43.25, TBil (μmol/L): 121.60 (83.20, 170.00) vs.15.25 (7.22, 29.05), DBil (μmol/L): 86.50 (58.60, 104.00) vs. 4.30 (2.22, 10.10), γ-GGT (U/L): 87.00 (37.00, 127.00) vs. 41.00 (19.00, 64.42), ALP (U/L): 199.10 (109.00, 349.20) vs. 125.50 (90.50, 155.25), TBA (μmol/L): 51.50 (16.20, 117.40) vs. 4.15 (2.02, 6.95), SCr (μmol/L): 155.80 (129.00, 237.00) vs. 79.00 (65.43, 113.70), INR: 1.28 (1.17, 1.63) vs. 0.94 (0.88, 1.08), all P < 0.05], prothrombin time (PT) was significantly prolonged [seconds: 16.10 (14.50, 19.20) vs. 12.40 (11.43, 13.40), P < 0.05]. The level of blood glucose (GLU), fibrinogen (FIB) and the activity of antithrombin Ⅲ (ATⅢ) decreased significantly [GLU (mmol/L): 5.18±1.33 vs. 6.33±1.19, FIB (g/L): 1.96±1.46 vs. 3.81±1.58, ATⅢ (%): 40.61±25.84 vs. 66.39±24.11, all P < 0.05]; ④ Complications: compared with HELLP syndrome group, the incidence of patients with hypoglycemia [30.77% (4/13) vs. 0% (0/34)], acute liver failure [53.85% (7/13) vs. 5.88% (2/34)], acute renal insufficiency [69.23% (9/13) vs. 8.82% (3/34)], coagulopathy [76.92% (10/13) vs. 38.24% (13/34)], disseminated intravascular coagulation (DIC) [53.85% (7/13) vs. 5.88% (2/34)], and multiple organ dysfunction syndrome (MODS) [53.85% (7/13) vs. 5.88% (2/34)] were significantly higher in AFLP group (all P < 0.05). ⑤ Maternal and neonatal outcome: all patients delivered after admission. The total length of hospital and intensive care unit stay were significantly longer in the AFLP group than in the HELLP syndrome group [days: 17.00 (11.00, 25.00) vs. 9.00 (7.00, 12.00), 12.00 (4.00, 22.00) vs. 3.91 (0, 7.00), both P < 0.01]. Two AFLP patients died, including one due to intracranial venous thrombosis and one due to multiple organ failure and cardiopulmonary arrest. There were no deaths in the HELLP syndrome group. Conclusions:There are significant differences in maternal characteristics, laboratory results and complications between AFLP and HELLP syndrome. TBil, γ-GGT, SCr, FIB, INR and ATⅢ activity may help to distinguish the two diseases.
7.Study on the production efficiency of platelet components in 24 prefecture-level blood stations in China
Minyu HUA ; Wei NIU ; Jian YAO ; Shouguang XU ; Yuxia QIU ; Li LI ; Dongmei ZHAO ; JiaYu WAN ; Feng YAN ; Hongzhi JIA ; Hao LI ; Jiaqi QIIAN ; Peng WANG ; Zhenxing WANG ; Lin BAO ; Shan WEN ; Sheng YE ; Xuefang FENG ; Man ZHANG ; Xiaobo CAI ; Wei ZHANG ; Dexu CHU ; Youhua SHEN ; Peifang CONG ; Hui ZHANG ; Yan QIU
Chinese Journal of Blood Transfusion 2022;35(9):937-942
【Objective】 To learn the production efficient of platelet components among prefecture-level blood stations in China, to provide supporting data for those blood stations to optimize the production mode of platelet components and continuously improve production efficiency and supply capacity. 【Methods】 The data from 2017 to 2020 was obtained from 24 prefecture-level blood stations who were the members of the practice comparison network for blood institutes in China. The collection units of apheresis platelets, the number of dual-collections of apheresis platelets and plasma, the average apheresis units of one platelet apheresis procedure, the discarded rate of apheresis platelets, the amount of expired apheresis platelets and the amount of apheresis platelets issued were collected. For concentrated platelets, the prepared amount of platelet concentrates and the amount of expired platelet concentrates were collected; both the quantity of qualified and issued concentrated platelets were submitted for statistical analysis.The total output and efficiency of platelet components were calculated based on the collected data. 【Results】 The average annual growth rate of apheresis platelets collection in 24 prefecture-level blood stations was 12.23%, accounting for 99.80% of the total platelet output; the average collection unit of one platelets apheresis procedure was 1.75; from 2019 to 2020, only 5 blood stations performed dual-collection of platelet and plasma during one apheresis procedure; the discarded rate of apheresis platelets was 0.28%, of which 0.007% was due to expiration. A total of 1 621.2 therapeutic units of concentrated platelets were prepared, and 13.03% of them was discarded due to the expiration. The production efficiency of platelet components was 97.56%, of which the production efficiency of apheresis platelets was 97.61% and the production efficiency of concentrated platelets was 74.43%. 【Conclusion】 There are large regional differences in the supply capacity of platelet components in prefecture-level blood stations. Apheresis platelets are the main resource of platelet components product, and the collection capacity is increasing over the years with the characteristics of high production efficiency and low expiration scrapping rate. However, the preparation of concentrated platelets are still limited with relatively low production and high expiration discarded rate.
8.An experimental study on the effect of different mechanical ventilation levels on right ventricular function in acute respiratory distress syndrome animals
Zidan WANG ; Rong LI ; Junfang LI ; Weigang LIU ; Dongmei SUN ; Yan XING ; Shizhong WANG ; Li YUAN ; Liang SHAN
Chinese Journal of Emergency Medicine 2020;29(3):365-372
Objective:To explore the relationship between respiratory mechanics and right heart function during ARDS mechanical ventilation through the establishment of Beagle dogs acute respiratory distress syndrome (ARDS) animal model and the application of different levels of mechanical ventilation, which will provide theoretical basis for right heart protective ventilation strategy of ARDS.Methods:Beagle dogs were anesthetized successfully and then pulmonary artery floating catheter, esophageal manometric catheter and femoral artery catheter were inserted. Under the pressure control mode, the driving pressure was fixed. After adjustment, PEEP gradually increased from 2 cmH 2O to 14 cmH 2O. The changes of respiratory mechanics, hemodynamics and right heart function were observed. ARDS model was established by injecting oleic acid into central vein, and mechanical ventilation with the same parameters was given after the model was established successfully. In contrast to itself, the changes of respiratory mechanics, hemodynamics and right heart function indexes of experimental dogs before and after modeling were analyzed. In the group, the indexes of different PEEP were compared by ANOVA, and then compared by Student-Newman-Keuls. The difference was statistically significant at a P value <0.05. Results:Before modeling, the peak airway pressure (P peak) and plateau pressure (P plat) increased with the increase of PEEP ( F=232.733,196.33, P<0.05). However, P trans-I, P trans-E, C stat and Vt decreased significantly ( F=4.524, 6.499, 64.803, 2.31, P<0.05). The area of change of right ventricle (FAC) became smaller ( F=3.09, P<0.05); SV first increased and then decreased ( F=3.24, P<0.05), and CVP and MPAP increased ( F=19.07,14.81, P<0.05). There was no significant difference in TAPSE, MAP, HR and SpO 2 ( P>0.05). After modeling, as PEEP increased, P peak, P plat, P ES-I and P ES-E increased significantly ( F=24.829, 41.95, 9.78, 87.86, P<0.05). Vt, P trans-I, P trans-E, C stat and Vt first increased and then decreased ( F=2.91, 4.29, 5.84, 48.890, P<0.05). TAPSE and SV first increased and then decreased ( F=6.22,6.54, P<0.05). CVP and MPAP increased ( F=5.23, 19.24, P<0.05). MAP increased first and then decreased ( F=5.02, P<0.05). SpO 2 increased ( F=2.77, P<0.05). FAC and HR had no statistical significance ( P>0.05). Conclusions:Trans pulmonary pressure and lung compliance can reflect the effectiveness of ARDS lung recruitment, and have good synergy; with the increase of PEEP, the right ventricular systolic function TAPSE is first affected, and SV compensatory increase, but with the increase of PEEP, TAPSE and SV decrease; pulmonary blood flow distribution is more important in improving alveolar oxygenation. Therefore, real-time monitoring of trans pulmonary pressure, TAPSE and intrapulmonary blood flow should be performed in ARDS treatment.
9.Internet behaviors and associated factor among Chinese primary school students
Chinese Journal of School Health 2020;41(5):704-708
Objective:
To analyze Internet behaviors of children and adolescents and their influencing factors, and to provide scientific basis for health education guidance and prevention measures.
Methods:
A stratified cluster sampling method was used. From January to June 2019, 30 955 primary school students in 16 provinces (municipalities), from eastern, central, and western economic zones of China were selected to conduct a questionnaire survey, regarding general demographics, diet and exercise, tobacco, alcohol and sleep, Internet and electronics usage, etc.
Results:
Among them, 43.1% of students had never been online with girls (44.7%) higher than boys (41.8%)(χ2=21.04, P<0.01); 1.3% of students reported online time for more than 4 hours a day, boys were significantly higher than girls(χ2=15.87, P<0.01); Internet addiction was detected in 2.4% of elementary school students, with boys (3.0%) higher than girls(1.7%)(χ2=331.77, P<0.01), and the trend increases with grade. With sixth grade (3.3%) were significantly higher than children from the first grade (0.8%) and other grades(χ2=163.96, P<0.01). Children’s tendency to Internet addiction was 11.9%, of which boys were 13.0% and girls were 10.6%, boys were significantly higher than girls(χ2=117.66, P<0.01). Pearson correlation analysis revealed that, after controlling gender age, boarding school, family relationship, single parents and regrouped families, smoking, drinking sugary drinks, eating fried foods, and left-behind were positively related to Intenet addiction(r=0.06, 0.07, 0.27, 0.44, 0.07, 0.11, 0.09, P<0.01), while medium and high intensity sports, effective physical education, health education in school, AIDS education and parental restrictions on TV and computer games, were negatively associated with internet addiction(r=-0.74, -0.65, -0.04, -0.05, -0.63, P<0.01).
Conclusion
The internet addiction of Chinese primary school students is common and increases by year. High-intensity, high-density sports, effective physical education, and health education could help prevent Internet addiction among children.
10.Upregulation of long non-coding RNA XIST has anticancer effects on epithelial ovarian cancer cells through inverse downregulation of hsa-miR-214-3p.
Changhong WANG ; Shan QI ; Cheng XIE ; Chunfu LI ; Pu WANG ; Dongmei LIU
Journal of Gynecologic Oncology 2018;29(6):e99-
OBJECTIVE: The present study is to evaluate the biological functions of long non-coding RNA (lncRNA), X-inactive specific transcript, X-inactive specific transcript (XIST) in human epithelial ovarian cancer (EOC). METHODS: XIST was upregulated in EOC cell lines, CAOV3 and OVCAR3 cells by lentiviral transduction. The effects of XIST overexpression on cancer cell proliferation, invasion, chemosensitivity and in vivo tumor growth were investigated, respectively. Possible sponging interaction between XIST and human microRNA hsa-miR-214-3p was further evaluated. Furthermore, hsa-miR-214-3p was overexpressed in XIST-upregulated CAOV3 and OVCAR3 cells to evaluate its effect on XIST-mediated EOC regulation. RESULTS: Lentivirus-mediated XIST upregulation had significant anticancer effects in CAOV3 and OVCAR3 cells by suppressing cancer cell proliferation, invasion, increasing cisplatin chemosensitivity and inhibiting in vivo tumor growth. Hsa-miR-214-3p was confirmed to directly bind XIST, and inversely downregulated in XIST-upregulated EOC cells. In EOC cells with XIST upregulation, secondary lentiviral transduction successfully upregulated hsa-miR-214-3p expression. Subsequently, hsa-miR-214-3p upregulation functionally reversed the anticancer effects of XIST-upregulation in EOC. CONCLUSION: Upregulation of lncRNA XIST may suppress EOC development, possibly through sponging effect to induce hsa-miR-214-3p downregulation.
Cell Line
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Cell Proliferation
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Cisplatin
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Down-Regulation*
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Humans
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MicroRNAs
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Neoplasm Invasiveness
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Ovarian Neoplasms*
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RNA, Long Noncoding*
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Up-Regulation*


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