1.Expression of serum exosomal LncRNA XIST and miR-130a-3p in gestational diabetes mellitus patients and its relationship with pregnancy outcome
Yu ZHAO ; Meifeng FANG ; Hu YUAN ; Yingna JIA ; Zhenyuan SONG
International Journal of Laboratory Medicine 2025;46(18):2177-2182
Objective To investigate the expression of serum exosomal long non-coding RNA X chromo-some inactivation specific transcript(LncRNA XIST)and microRNA(miR)-130a-3p in gestational diabetes mellitus(GDM)patients and its relationship with pregnancy outcome.Methods A total of 149 GDM patients(GDM group)and 149 healthy pregnant women(control group)admitted to the hospital from January 2021 to January 2024 were selected.Serum exosomal LncRNA XIST,miR-130a-3p and glucose and lipid metabolism indexes of the two groups were detected and compared.The correlation of LncRNA XIST and miR-130a-3p levels with glycolipid metabolism indexes was analyzed by Pearson method.Univariate and multivariate Logis-tic regression models were used to analyze the influencing factors of pregnancy outcomes in GDM patients.Re-ceiver operating characteristic(ROC)curve was used to analyze the predictive efficacy of serum exosomal Ln-cRNA XIST and miR-130a-3p levels alone and in combination for pregnancy outcomes in GDM patients.Results Serum exosomal LncRNA XIST levels,fasting insulin(FINS),fasting blood glucose(FBG),insulin resistance index(HOMA-IR),2-hour postprandial glucose(2 hPG),total cholesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)levels in GDM group were higher than those in control group(P<0.05),and the levels of miR-130a-3p and high density lipoprotein cholesterol(HDL-C)were lower than those in the control group(P<0.05).LncRNA XIST was positively correlated with FINS,FBG,HOMA-IR,2 hPG,TC,TG and LDL-C,and negatively correlated with HDL-C(P<0.05).miR-130a-3p was negatively correlated with FINS,FBG,HOMA-IR,2 hPG,TC,TG and LDL-C,and positively correlated with HDL-C(P<0.05).Among the 149 GDM patients,43 cases showed adverse pregnancy outcome,accounting for 28.86%.The increase level of FBG,the increase level of LncRNA XIST and the increase of HOMA-IR were risk factors for adverse pregnancy outcome in GDM patients,while the increase of miR-130a-3p was a protec-tive factor.The ROC curve analysis showed that the predictive efficacy of combined detection was superior to that of single indicator detection.Conclusion The serum exosomal LncRNA XIST of GDM patients is highly expressed,and miR-130a-3p is lowly expressed,and the combined detection of the above indexes has a high predictive value for pregnancy outcome.
2.FBXW7 promotes ferroptosis in head and neck squamous cell carcinoma cells through inhibiting c-Myc/SOX2/SLC7A11
Yiren CHEN ; Zhenyuan ZHAO ; Yangyu ZHENG ; Wei ZHANG ; Xiaomeng SONG
STOMATOLOGY 2024;44(6):426-432
Objective To explore the effect of FBXW7 on ferroptosis in head and neck squamous cell carcinoma.Methods Head and neck squamous cell lines HN4 and HN6 were cultured in vitro.FBXW7 and SOX2 overexpression plasmids were constructed,and the plasmids were stably transfected into cell lines.The overexpression transfection efficiency was verified at the transcription level and protein level by qRT-PCR and Western blot experiments,respectively.The lipid peroxidation levels of head and neck squamous cell carcinoma cells with overexpressing FBXW7 were verified by measuring malondialdehyde(MDA),glutathione(GSH),and reactive ox-ygen species(ROS)levels.After treating cells with ferroptosis inhibitor Fer-1,the changes in cell viability were further detected to ver-ify the effect of FBXW7 on ferroptosis.The effect of transfection of the overexpressed plasmid on cellular pathways was detected by Western blot.Results HN4 and HN6 cell lines showed increased levels of lipid peroxidation after overexpression of FBXW7,and the ferroptosis inhibitor Fer-1 was able to effectively reverse the ferroptosis induced by overexpression of FBXW7.Western blot assay results showed that overexpression of FBXW7 reduced the expression of c-Myc,SOX2 and SLC7A11.Conclusion FBXW7 regulates the ex-pression of SOX2-SLC7A11 by degrading c-Myc,thereby effectively regulating ferroptosis in HNSCC.
3.The effect of knocking down Sec31A on the malignant phenotype of HNSCC
Yao HE ; Zhenyuan ZHAO ; Teng GAO ; Peng LIN ; Yiren CHEN ; Xiaomeng SONG
STOMATOLOGY 2024;44(7):487-493
Objective To explore the impact of knocking down Sec31A on the malignant phenotype of head and neck squamous cell carcinoma(HNSCC)and its possible mechanisms.Methods Transcriptome sequencing data of HNSCC tissues and adjacent tissues were obtained from the TCGA database,and the expression levels of Sec31A were compared.Immunohistochemical staining was used to analyze the expression of Sec31A in HNSCC tissues.Kaplan-Meier survival analysis was used to compare the relationship between Sec31A and the prognosis of HNSCC patients.Small interfering plasmids si-Sec31A and si-NC were transfected into HNSCC cell lines HN6 and HN4,and the impact of knocking down Sec31A on the biological behavior of HNSCC cells was detected through CCK-8 exper-iments,plate cloning experiments,scratch healing experiments,and Transwell experiments.Changes in the expression levels of PI3K/AKT/mTOR pathway related proteins in cells were detected after knocking down Sec31A with HN6 and HN4 through Western Blot(WB)experiments.Stable transfected cell lines of HN6 siSec31A and HN6 siNC were constructed and inoculated subcutaneously in nude mice to further verify the tumorigenic effect of Sec31A in vivo.Results TCGA data showed that Sec31A was higher in HNSCC tissues than in adjacent normal tissues(P<0.01),and high expression of Sec31A was significantly correlated with poor prognosis in pa-tients(P<0.05).Immunohistochemical staining showed that Sec31A was expressed stronger in HNSCC tissues than in normal tissues.In HN6 and HN4 cells,knocking down Sec31A resulted in significantly weaker proliferation,migration,and invasion abilities compared to the control group.Through WB experiments,it was found that transfection of si-Sec31A with HN6 and HN4 significantly reduced the expression levels of p-PI3K,p-AKT,and p-mTOR proteins.After knocking down Sec31A with HN6,the transplanted tumor volume in nude mice was significantly smaller than that in the control group.Conclusion Knocking down Sec31A can inhibit the proliferation,migration and invasion of HNSCC cells,possibly through the PI3K/AKT/mTOR pathway.
4.Value of peritoneal protein clearance as a predictor of cardiovascular outcomes in peritoneal dialysis patients
Wei NIU ; Xiaoxiao YANG ; Yiwei SHEN ; Dahua MA ; Yimei XU ; Qianhui SONG ; Zanzhe YU ; Hao YAN ; Zhenyuan LI ; Zhaohui NI ; Wei FANG
Chinese Journal of Nephrology 2021;37(7):576-582
Objective:To investigate the predictive value of peritoneal protein clearance (Pcl) for cardiovascular events and cardiovascular mortality in peritoneal dialysis (PD) patients.Methods:Eligible PD patients were prospectively enrolled from January 2014 to April 2015 in the PD Center of Renji Hospital, School of Medicine, Shanghai Jiao Tong University. All patients were followed up until death, withdrawing from PD, transferring to other centers, or the end of study period (October 1, 2018). The patients were divided into high Pcl group and low Pcl group by the median Pcl, and the differences of related indicators between the two groups were compared. A multiple linear regression model was used to analyze the influencing factors of Pcl. The Kaplan-Meier method and Log-rank test were used to compare the cumulative survival rates of patients between the two groups. A multivariate Cox regression model was used to estimate the risk of cardiovascular events and cardiovascular mortality in relation to Pcl in PD patients.Results:A total of 271 patients were enrolled, with 135 males (49.8%), age of (56.92±0.84) years old and a median PD duration of 38.77(19.00, 63.10) months. There were 70 patients (25.8%) comorbiding with diabetes and 81 patients (29.9%) with cardiovascular diseases (CVD). The median Pcl of this cohort was 67.93(52.31, 88.36) ml/d. Compared with the low Pcl group (Pcl<67.93 ml/d), the high Pcl group (Pcl≥67.93 ml/d) had older age, and greater proportion of CVD, body mass index (BMI), pulse pressure, brain natriuretic peptide, mass transfer area coefficient of creatinine (MTACcr), and lower serum albumin (all P<0.05). There was no significant difference in gender, dialysis duration, proportion of diabetes, proportion of angiotensin converting enzyme inhibitor and angiotensin receptor blocker, proportion of continuous ambulatory PD, high sensitivity C reactive protein, fluid removal including 24 h urine volume and 24 h ultrafiltration, and residual renal function between the two groups (all P>0.05). Multiple linear regression analysis showed that serum albumin ( β=-0.388, P<0.001), BMI ( β=0.189, P<0.001), and MTACcr ( β=0.247, P<0.001) were independently related to lg(Pcl). During the study period, 55 patients experienced one or more cardiovascular events and 39 patients had cardiovascular mortality. According to Kaplan-Meier analysis, cardiovascular mortality in the high Pcl group was higher than that of low Pcl group (Log-rank χ2=6.902, P=0.009). Multivariate Cox regression analysis showed that, high lg(Pcl) was an independent influencing factor of cardiovascular events in PD patients ( HR=7.654, 95% CI 1.676-34.945, P=0.009). Conclusions:Serum albumin, BMI and MTACcr are independently associated with Pcl, and Pcl is an independent predictor of cardiovascular events in PD patients.
5.Expression of miR-520d-3p in X-rays irradiated A549 cells and serum samples of non-small cell lung cancer patients and its significance
Xiujun SONG ; Qisheng JIANG ; Sinian WANG ; Huijie YU ; Xiao LI ; Zhenyuan LI ; Jin LYU
Cancer Research and Clinic 2017;29(7):433-436,441
Objective To investigate the expression of miR-520d-3p in 2 and 4 Gy X-ray irradiated A549 cells, serum samples of non-small cell lung cancer (NSCLC) patients and its significance. Methods Real time quantitative PCR (RT-qPCR) was employed to detect the expression miR-520d-3p in 2 and 4 Gy X-ray irradiated A549 cells in vitro. Besides, 0, 2 and 4 Gy X-ray irradiated A549 cells were used to prepare animal model of nude mice lung metastasis through mainline. The expressions of miR-520d-3p and lung tissue of nude mice were detected, and the serum samples of NSCLC patients were collected to test the expression of miR-520d-3p . Results Compared with the control group ( 0 Gy ) , the expression of miR-520d-3p was up-regulated significantly in 2 and 4 Gy X-ray irradiated A549 cells after 48 hours (1.00±0.03, 1.47±0.10, 1.84 ±0.09 respectively), and there was a significant difference (F= 94.350, P= 0.000). Furthermore, compared the control group with injection after 10 weeks, the expressions of miR-520d-3p in nude mice lung tissue in 0, 2 and 4 Gy X-ray irradiated groups were increased (2.33 ±0.13, 8.24 ±0.25, 3.46 ±0.14, respectively) (F=1.787, P= 0.227), and the expressions in serum were increased (11.43±2.30, 10.22±4.62, 8.99 ±3.67, respectively) (F= 1.547, P= 0.246). Out of 20 serum samples of NSCLC patients (including 10 cases of adenocarcinoma, 10 cases of squamous carcinoma ), 11 cases (55%) were detected with up-regulated miR-520d-3p expression. Conclusions 2 and 4 Gy X-ray could increase the expression of miR-520d-3p of A549 cells in vitro and in vivo, which might be correlated with the enhanced invasion and metastasis of A549 cells induced by X-ray in vitro and in vivo. Furthermore, the expressions of miR-520d-3p were increased significantly in over 50%serum samples of NSCLC patients, which might be a new marker for the diagnosis of NSCLC.
6.The expression of miR-424* in vivo and in vitro irradiated A549 cells, tissue and serum samples of non-small cell lung cancer
Jun GAO ; Jin LYU ; Bin HU ; Xiujun SONG ; Ying DUAN ; Zhenyuan LI ; Xiao LI ; Lina YANG ; Sinian WANG ; Qisheng JIANG
Chinese Journal of Radiological Medicine and Protection 2017;37(5):332-338
Objective To investigate the expression of miR-424* in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of non-small cell lung cancer(NSCLC) patients,and to explore its potential role in the diagnosis and prognosis of lung cancer.Methods A549 cells were irradiated with 2 and 4 Gy X-rays,and some of irradiated cells were injected into nude mice through tail vein.Real time quantitative PCR (RT-qPCR) assay was employed to detect the expression of miR-424 * in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of lung cancer patients.Results Compared with the control group,the expression of miR-424* was up-regulated significantly in X-ray irradiated A549 cells at 1,2,12,24 and 48 hpost irradiation,respectively (2 Gy:t =-45.886--6.709,P <0.05;4 Gy:t =-29.087--7.833,P < 0.05).Furthermore,the expression of miR-424 * was up-regulated in the lung and serum of nude mice with injection of 0,2 and 4 Gy X-ray irradiated A549 cells,compared with control group (fold change was 9.72,8.58 and 4.7 with 2 Gy irradiation and 11.93,9.22 and 8.99 with 4 Gy irradiation,t=-13.243,-12.409,-9.833 in lung andt=-6.436,-3.052,-3.609 in serum,respectively,P < 0.05).Out of 11 tissue samples of NSCLC patients,6 were detected with up-regulated miR-424* expression,and no significant discrepancy of miR-424* expression was detected in two type of NSCLC tissue samples.On the contrary,43 serum samples were detected with up-regulated miR-424* expression out of 84 serum samples (51.20%) of NSCLC patients (fold change range 1.97 to 17.71),and significant discrepancy of miR-424* expression was shown in two subtypes of NSCLC serum samples [adenocarcinoma:39.10% (18/46) and squamous carcinoma:65.8% (25/38)],as well as in serum samples of NSCLC patients with radiotherapy [41.5% (22/53)] and without radiotherapy [67.7% (21/31)] (t=5.919,5.387,P <0.05,respectively).Conclusions 2 and 4 Gy X-ray irradiation could up-regulate the expression of miR-424* in A549 cells,which might be correlated with the enhanced metastasis of A549 cells induced by X-ray in vivo and in vitro.Furthermore,the expression of miR-424* was up-regulated in over 50% of the tissue and serum samples of NSCLC patients,which might be correlated with the diagnosis of NSCLC subtype and prognosis of radiotherapy.
7.Comparison the effect of surgical and conservative treatment of acute dislocation
Huixin SONG ; Xiangli LI ; Wei ZHAO ; Zhifeng LI ; Zhenyuan ZHAO ; Baozhong LI ; Zhiguo LI
Clinical Medicine of China 2013;29(8):858-861
Objective To compare the clinical results of surgical and conservative treatment of acute patellar dislocations.Methods Retrospective analyzed the clinical data of 35 patients with acute patellar dislocations from June 2004 to October 2009,and divided the patients into 2 groups.One group with 18 patients underwent surgical treatments,and the other group with 17 patients underwent conservative treatments,record the number of relapses cases of both the two groups.A radiographic examination was performed in the evaluation of the patients,and the Kujala questionnaire was applied with the intention of analyzing the improvement of pain and quality of life.Results All patients were followed up for more than 12 months.(1) The conservative treatment group exhibited a higher number of recurrent dislocations (7 patients) (41.2%) than the surgical treatment group,which did not have any relapses (x2 =9.265,P =0.002).(2) The patellar tilt returned to normal in the surgical group,while 8 patients returned to normal in the conservative group,with statistical difference between groups (x2 =10.980,P =0.001).And the lateral shift ratio returned to normal in the surgical group,while 6 patients returned to normal in the conservative group,with statistical difference between groups(x2 =7.667,P =0.006).(3)The surgical treatment group obtained a significantly better mean score on the Kujala test than preoperative ((90 ± 5) vs.(58 ± 6),t =16.465,P < 0.01) ; The conservative treatment group,compared with the preoperative,is not improved obviously ((72 ± 6) vs.(62 ± 8),t =0.943,P > 0.05) ;Postoperative group comparison,surgical group was higher than that in conservative group,there was significant difference between two groups (t =12.256,P < 0.01).Conclusion For acute patellar dislocation,surgical treatment can significantly restore patellar stability,improve the function of knee joint.
8.Metabolism of mangiferin by human intestinal bacteria in vitro.
Huixue HUANG ; Zhenyuan TAN ; Jiagang DENG ; Qiuyun LIANG ; Yumei NONG ; Nianmei SONG
China Journal of Chinese Materia Medica 2011;36(4):443-445
OBJECTIVETo study the metabolism of mangiferin by human intestinal bacteria in vitro.
METHODHuman intestinal bacteria and mangiferin were incubated under anaerobic conditions in vitro. The metabolite was separated and purified by D101 macroporous resin column and preparation high performance liquid chromatography, and its structure was identified by MS and NMR.
RESULTAfter 12 h incubation with human intestinal bacteria, the content of mangiferin metabolite reached the maximum, and it was determined as 1, 3, 6, 7-tetrahydroxyxanthen by MS and NMR.
CONCLUSIONMangiferin can be metabolized in vitro by human intestinal bacteria into its aglycone (1, 3, 6, 7-tetrahydroxyxanthen).
Bacteria ; metabolism ; Chromatography, High Pressure Liquid ; Humans ; Intestines ; microbiology ; Xanthones ; metabolism

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