1.Effects of inhibition of glucose-6-phosphate dehydrogenase expression on proliferation and cell cycle distribution of A431 cells
Min LI ; Yonghua XIA ; Dong LIU ; Mengjie ZHANG ; Zhongwei TIAN ; Zhanguo LI
Chinese Journal of Dermatology 2016;49(11):766-770
Objective To evaluate effects of downregulation of glucose?6?phosphate dehydrogenase(G6PD) expression on proliferation and cell cycle distribution of cutaneous squamous cell carcinoma(CSCC)cells. Methods Western blot analysis was performed to measure the protein expression of G6PD in normally cultured human HaCaT keratinocytes, SCL?1 and A431 CSCC cells. When A431 cells grew to 85%-90%confluence, a small interfering RNA (siRNA)targeting G6PD(G6PD?siRNA group)and a negative control siRNA(siRNA control group)were transfected into them separately, and untransfected A431 cells served as the untransfected group. CCK?8 assay was performed to evaluate proliferative activity of the A431 cells on days 0, 1, 2, 3 and 4 after transfection, Western blot analysis to measure G6PD, cyclin D1 and CDK4 protein expressions in A431 cells, and flow cytometry to analyze cell cycle distribution in A431 cells after 48 hours of additional culture. Results The protein expression of G6PD was significantly higher in normally cultured SCL?1 cells(0.308 ± 0.023)and A431 cells(0.643 ± 0.046)than in HaCaT cells(0.100 ± 0.019, both P<0.05), and significantly higher in A431 cells than in SCL?1 cells(P<0.05). The G6PD?siRNA group showed significantly decreased protein expressions of G6PD, cyclin D1 and CDK4(0.134 ± 0.027, 0.154 ± 0.017 and 0.166 ± 0.017, respectively)compared with the untransfected group(0.425 ± 0.029, 0.344 ± 0.024 and 0.330 ± 0.020 respectively)and siRNA control group(0.444 ± 0.033, 0.350 ± 0.027 and 0.348 ± 0.018 respectively) (all P<0.05). Besides, the G6PD?siRNA group showed significantly decreased cellular proliferative activity on days 1-4 compared with the siRNA control group and untransfected group(all P<0.001), while there were no significant differences between the untransfected group and siRNA control group at any of the time points (all P > 0.05). Compared with the untransfected group and siRNA control group, the G6PD?siRNA group showed significantly higher proportions of A431 cells in G0/G1 phase(both P < 0.001), but significantly lower proportions of A431 cells in S phase(both P<0.001). Conclusion G6PD may play important roles in the regulation of proliferation and cell cycle distribution of CSCC cells.
2.Visualization of Multivariate Metabolomic Data
Jun ZHOU ; Jiye AA ; Guangji WANG ; Fengyi ZHANG ; Rongrong GU ; Xinwen WANG ; Chunyan ZHAO ; Mengjie LI ; Jian SHI ; Bei CAO ; Tian ZHENG ; Linsheng LIU ; Sheng GUO ; Jinao DUAN
Chinese Herbal Medicines 2011;(4):285-289
Objective Although principal components analysis profiles greatly facilitate the visualization and interpretation of the multivariate data,the quantitative concepts in both scores plot and loading plot are rather obscure.This article introduced three profiles that assisted the better understanding of metabolomic data.Methods The discriminatory profile,heat map,and statistic profile were developed to visualize the multivariate data obtained from high-throughput GC-TOF-MS analysis.Results The discriminatory profile and heat map obviously showed the discriminatory metabolites between the two groups,while the statistic profile showed the potential markers of statistic significance.Conclusion The three types of profiles greatly facilitate our understanding of the metabolomic data and the identification of the potential markers.
3. Effect of epigallocatechin gallate on Th1/Th2 immune balance in patients with plaque-type psoriasis
Dandan FU ; Hua HU ; Mengjie ZHANG ; Min LI ; Zhanguo LI ; Zhongwei TIAN
Chinese Journal of Dermatology 2019;52(9):611-615
Objective:
To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.
Methods:
A total of 33 patients with plaque-type psoriasis vulgaris were enrolled, and peripheral blood mononuclear cells (PBMC) were isolated and cultured. The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay. PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours. Flow cytometry was performed to determine proportions of Th1 and Th2 cells, enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin[IL]-2, interferon[IFN]-γ) and Th2 cytokines (IL-4, IL-10) in the cell culture supernatant, and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor) . Statistical analysis was carried out by using
4.Analysis of clinical factors on early pregnancy loss during in vitro fertilization-embryo transfer treatment and natural pregnancy
Mengjie FAN ; Chan TIAN ; Liang CHANG ; Xiaoye WANG ; Rong LI
Chinese Journal of Obstetrics and Gynecology 2018;53(11):755-760
Objective To compare differences of clinical factors related to early pregnancy loss between invitro fertilization-embryo transfer (IVF-ET) treatment and natural pegnancy. Methods A retrospective analysis was performed on the 363 cases of early pregnancy loss between Dec. 2015 to May 2016 in Peking University Third Hospital, during which 173 cases were after IVF-ET treatment(IVF-ET group), and others were natural pregnancies(natural group). Results The average age in IVF-ET group was significantly higher than that in the natural group [(34.1±4.3)versus(31.8±4.1)years old, P<0.01]. The terminating time of pregnancy loss in IVF-ET group was short than that in the natural group [(59.8±9.2) versus(69.9 ± 11.1)days, P<0.01]. The incidence of embryo abnormal chromosome in IVF-ET group was significantly lower than that in the natural group [57.2%(99/173)versus 74.2%(141/190), P<0.01], during which abnormal chromosome numbers were the most common. Conclusions The pregnancy loss of early pregnancy is mainly caused by chromosome abnormality. The proportion of chromosome abnormality in early pregnancy loss after IVF-ET is not higher than that of natural pregnancy, indicating that there are relatively reliable gametes and embryo safety in IVF treatment.
5.Subtype discrimination of lung adenocarcinoma manifesting as ground glass nodule based on radiomics
Li FAN ; Mengjie FANG ; Di DONG ; Wenting TU ; Yun WANG ; Qiong LI ; Yi XIAO ; Jie TIAN ; Shiyuan LIU
Chinese Journal of Radiology 2017;51(12):912-917
Objective To develop and validate the radiomics nomogram on the discrimination of lung invasive adenocarcinoma from'non-invasive'lesion manifesting as ground glass nodule(GGN)and compare it with morphological features and quantitative imaging. Methods One hundred and sixty pathologically confirmed lung adenocarcinomas from November 2011 to December 2014 were included as primary cohort. Seventy-six lung adenocarcinomas from November 2014 to December 2015 were set as an independent validation cohort. Lasso regression analysis was used for feature selection and radiomics signature building. Radiomics score was calculated by the linear fusion of selected features. Multivariable logistic regression analysis was performed to develop models. The prediction performances were evaluated with ROC analysis and AUC,and the different prediction performance between different models and mean CT value were compared with Delong test. The generalization ability was evaluated with the leave-one-out cross-validation method. The performance of the nomogram was evaluated in terms of its calibration. The Hosmer-Lemeshow test was used to evaluate the significance between the predictive and observe values.Results Four hundred and eighty-five 3D features were extracted and reduced to 2 features as the most important discriminators to build the radiomics signatures. The individualized prediction model was developed with age, radiomics signature, spiculation and pleural indentation, which had the best discrimination performance(AUC=0.934)in comparison with other models and mean CT value(P<0.05)and showed better performance compared with the clinical model(AUC=0.743,P<0.001).The radiomics-based nomogram demonstrated good calibration in the primary and validation cohort, and showed improved differential diagnosis performance with an AUC of 0.956 in the independent validation cohort. Conclusion Individualized prediction model incorporating with age, radiomics signature, spiculation and pleural indentation, presenting with radiomics nomogram, could differentiate IAC from'non-invasive'lesion manifesting as GGN with the best performance in comparison with morphological features and quantitative imaging.
6.SNHG6 promotes invasion and metastasis of esophageal squamous cell carcinoma TE1 cells via upregulating ZEB1
WANG Mengjie ; LIU Yan ; LYU Wei ; TIAN Cong ; WANG Yu ; ZHAO Lianmei ; LIU Lihua
Chinese Journal of Cancer Biotherapy 2020;27(3):282-288
Objective: To explore the roles and mechanisms of long non-coding RNA (lncRNA) small nucleolar RNA host gene 6 (SNHG6) in promoting invasion and metastasis of esophageal squamous carcinoma (ESCC). Methods: Real time quantitative polymerase chain reaction (qPCR) was used to detect the expression of SNHG6 in ESCC and matched para-carcinoma tissues. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the expression of SNHG6 in ESCC cell lines (TE1, Yes-2, Eca9706 and Kyse150). Then, TE1 cell line which harbored highest expression of SNHG6 was used in following experiments. siRNAs were used to knock down the expression of SNHG6. Clone formation, wound-healing and transwell assay were used to detect the abilities of proliferation, migration andinvasionofTE1cells,respectively.Westernblottingwasusedtodetecttheexpressions of MMP-2, MMP-9andZEB1 protein before and after knockdownofSNHG6inTE1cells.Results:SNHG6washighlyexpressedinESCC tissues, compared to para-carcinoma tissues (P<0.01). The expression of SNHG6 was significantly decreased after transfection of SNHG6siRNA (all P<0.01). The abilities of proliferation, migration and invasion of TE1 cells in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.01). The expressions of ZEB1, MMP-2and MMP-9 in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.05). Conclusion: SNHG6 is highly expressed in ESCC tissues and promotes the malignant biological behavior of ESCC cells. Its mechanism of promoting the occurrence and development of ESCC may be related to the upregulation of ZEB1 expression.
7.Critical effects of long non-coding RNA on fibrosis diseases
Yue ZHANG ; Gang LUO ; Yi ZHANG ; Mengjie ZHANG ; Jian ZHOU ; Weiwu GAO ; Xiuyun XUAN ; Xia YANG ; Di YANG ; Zhiqiang TIAN ; Bing NI ; Jun TANG
Experimental & Molecular Medicine 2018;50(1):e428-
The expression or dysfunction of long non-coding RNAs (lncRNAs) is closely related to various hereditary diseases, autoimmune diseases, metabolic diseases and tumors. LncRNAs were also recently recognized as functional regulators of fibrosis, which is a secondary process in many of these diseases and a primary pathology in fibrosis diseases. We review the latest findings on lncRNAs in fibrosis diseases of the liver, myocardium, kidney, lung and peritoneum. We also discuss the potential of disease-related lncRNAs as therapeutic targets for the clinical treatment of human fibrosis diseases.
Autoimmune Diseases
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Fibrosis
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Genetic Diseases, Inborn
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Humans
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Kidney
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Liver
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Lung
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Metabolic Diseases
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Myocardium
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Pathology
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Peritoneum
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RNA, Long Noncoding
8.Meta-analysis of associations between psoriasis and adverse pregnancy outcomes
Shasha ZHANG ; Xiangfeng SONG ; Min LI ; Dandan FU ; Mengjie ZHANG ; Hua HU ; Zhongwei TIAN
Chinese Journal of Dermatology 2020;53(4):299-302
Objective:To evaluate associations of psoriasis with adverse pregnancy outcomes.Methods:Databases, including CNKI, Wanfang, VIP, PubMed, Embase and Cohrane Library databases, were searched for published articles on associations between psoriasis and adverse pregnancy outcomes between January 1980 and December 2018. Quality of included articles was assessed by using MOOSE checklist. Statistical analysis was carried out with Review Manager 5.3 software.Results:One cohort study, 4 case-control studies and 2 cross-sectional studies meet the inclusion criteria. After heterogeneity test, meta-analysis was carried out by using a random effect model. The risks of cesarean ( OR= 1.17, 95% CI: 1.01- 1.37) , eclampsia or preeclampsia ( OR= 1.34, 95% CI: 1.00- 1.79) and premature birth ( OR= 1.09, 95% CI: 1.00- 1.19) were significantly higher in patients with psoriasis than in individuals without psoriasis (all P < 0.05) . There was no significant difference between patients with psoriasis and individuals without psoriasis in the risks of spontaneous abortion, low birth weight, high birth weight, stillbirth, congenital malformation, placental abruption, overdue delivery, low Apgar score (< 7) , polyhydramnios and oligohydramnios. Conclusion:The risks of cesarean, eclampsia or preeclampsia, and premature birth are higher in patients with psoriasis than in individuals without psoriasis.
9.Study on effects of Jianpi Qinghua Decoction on hepatocyte apoptosis in nonalcoholic fatty liver disease model mice
Xu HAN ; Mengjie CAI ; Qingguang CHEN ; Jing TIAN ; Hao LU
International Journal of Traditional Chinese Medicine 2023;45(11):1382-1385
Objective:To study the effects of Jianpi Qinghua Decoction on hepatocyte apoptosis under non-alcoholic fatty liver disease (NAFLD), and to discuss its mechanism.Methods:Totally 29 C57 mice were randomly selected and fed a 60% high fat diet for 16 weeks, while the remaining 6 mice were given regular feed as the normal group. After successful modeling, 12 mice with larger body weight were divided into TCM group and model group using a random number table method, while continuing to receive high-fat feed. The TCM group was orally administered Jianpi Qinghua Decoction 20.961 g/kg. The normal group and model group were orally administered an equal volume of distilled water once a day, with continuous intervention for 4 weeks. The levels of GOT and GPT in blood were detected by ELISA, the deposition of triglyceride in liver was detected by oil red O, and the apoptosis of liver cells was detected by TUNEL fluorescence staining. Western blot was used to detect the expressions of cleaved Caspase-3, Caspase-3, Bax, Bcl-2, JNK and p-JNK.Results:Compared with the model group, the body weight and GPT in the TCM group significantly decreased ( P<0.05), TG deposition was significantly reduced, apoptosis range of liver cells was significantly reduced, and cleaved Caspase-3/Caspase-3, p-JNK/JNK and the expression of Bax significantly decreased ( P<0.05). The expression of Bcl-2 increased ( P<0.05). Conclusion:Jianpi Qinghua Decoction can inhibit JNK protein phosphorylation and effectively reduce liver cell apoptosis in NAFLD mice, which may delay the progression of NAFLD towards cirrhosis and liver cancer.
10.lncRNA NEAT1 promotes cell proliferation of lung adenocarcinoma PC-9 cells through inhibiting DNAdamage
LIU Tianxu ; WANG Mengjie ; TIAN Cong ; LIU Yan ; LYU Wei ; JIA Yunlong ; LIU Lihua
Chinese Journal of Cancer Biotherapy 2019;26(8):845-849
Objective: To investigate the effects of long non-coding RNA nuclear enriched abundant transcript 1 (lncRNA NEAT1) on the proliferation of lung adenocarcinoma PC-9 cells and to explore its mechanism. Methods: qPCR was used to detect the expression level of lncRNA NEAT1 in human lung adenocarcinoma PC-9 cells and human embryonic lung diploid 2BS cells. The sequence of small interfering RNA(siRNA) targeting lncRNANEAT1 gene was designed and synthesized, and then transfected into PC-9 cells by liposome method. The expression level of NEAT1 in PC-9 cells before and after transfection was detected by qPCR. MTT and flow cytometry were used to detect the effect of lncRNANEAT1 knockdown on proliferation and cell cycle distribution of PC-9 cells, respectively. WB assay was used to detect the expressions of DNA damage-related proteins, namely, double-stranded DNA breaks (DSBs) biomarker γ-H2AX and ataxia-telangiectasia mutated (ATM), before and after transfection. Results: Compared with 2BS cells, lncRNA NEAT1 was highly expressed in PC-9 cells (P<0.05). The PC-9 cells with lncRNA NEAT1 knock-down were successfully established. After being transfected with siRNA for 12 h, the proliferation of PC-9 cells in siNEAT1 group and siNEAT2 group significantly decreased as compared with the blank control group and the empty transfection group (P<0.05). In the interference groups, cell cycle was arrested in G1 phase ([88.97±2.64]%, [88.15±1.48]% vs [84.5±1.72]%, P<0.05) and G2/M phase ([8.35±2.02]%, [8.11± 1.36]% vs [4.28±1.28]%, P<0.05). The expression levels of DNA damage-related proteinsATM and γ-H2AX in the interference groups were significantly increased (all P<0.05). Conclusion: lncRNA NEAT1 is highly expressed in lung adenocarcinoma PC-9 cells. lncRNA NEAT1 inhibits DNA damage and causes cell cycle at G1/M phase switch, and thus promotes the proliferation of lung adenocarcinoma cells.