1.The role and mechanism of long non-coding RNA KCNQ1 overlapping transcript 1 in the proliferation, migration and invasion of hepatocellular carcinoma
Hongliang MEI ; Zhiyuan HUANG ; Yilin HU ; Yan JIANG ; Min CHEN ; Qiping LU ; Zhisu LIU
Chinese Journal of Digestive Surgery 2020;19(10):1083-1090
Objective:To explore the role and mechanism of long non-coding RNA KCNQ1 overlapping transcript 1 (LncRNA KCNQ1OT1) in the migration, proliferation and invasion of hepatocellular carcinoma (HCC).Methods:The experimental method was conducted. The expression levels of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues in the StarBase database were collected. The experimental methods including real-time quantitative PCR, cell transfection, scratch assay, CCK8 assay, Transwell assay, Western blot were used to determine the expression, migration, proliferation, invasion of LncRNA KCNQ1OT1 in HCC cells and its relationship with phosphatidylinositol 3-kinase/phosphorylated AKT Protein (PI3K /p-AKT) signaling pathways. Observation indicators: (1) expression of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues; (2) the migration of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown; (3) the proliferation and invasion of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown; (4) effects of LncRNA KCNQ1OT1 gene knockdown on PI3K/p-AKT signaling pathways. Measurement data with normal distribution were expressed as Mean± SD, and comparison between groups was analyzed using the t test. The Kaplan-Meier method was used to calculate survival rates and draw survival curves. Results:(1) Expression of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues. The expression levels of LncRNA KCNQ1OT1 in 374 HCC tissues and 50 normal liver tissues from StarBase database were 3.320±0.017 and 1.470±0.025, respectively, showing a significant difference ( t=5.24, P<0.05). Results of gene expression profile interactive analysis showed that the 30-month disease-free survival rates of HCC patients with high and low expression levels of LncRNA KCNQ1OT1 were 41% and 55%, respectively, with a significant difference ( χ2=6.209, P<0.05). The relative expression of LncRNA KCNQ1OT1 in HepG2, SMCC-7721and MHCC-97H cells were 1.470±0.042, 3.300±0.032, 4.040±0.031, respectively, versus 1.000±0.022 in normal liver cells (LO2), showing significant differences ( t=17.66, 95.40, 114.20, P<0.05). (2) The migration of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown. ① Results of cell transfection showed that the relative expression levels of LncRNA KCNQ1OT1 in HepG2, SMCC-7721 and MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.350±0.016, 0.310±0.020, 0.380±0.018, respectively, versus 1.000±0.021, 1.000±0.018, 1.000±0.019 in the negative control cells, showing significant differences ( t=23.40, 28.15, 22.32, P<0.05). ② Results of scratch assay showed that the healing rates of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 85.0%±1.9%, 75.0%±1.8%, 90.0%±1.7%, respectively, versus 100.0%±2.0%, 95.0%±1.8%, 72.0%±1.7% of the negative control cells, showing significant differences ( t=31.35, 47.36, 38.42, P<0.05). ③ Results of Transwell assay showed that the vertical migration rates of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 195±10, 205±12, 85±8, respectively, versus 520±11, 430±7, 405±20 of the negative control cells, showing significant differences between them ( t=922.30, 458.20, 708.40, P<0.05). (3) The proliferation and invasion of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown. ① Results of CCK8 assay showed that 72-hour optical densities of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 1.370±0.018, 1.240±0.016, 1.360±0.020, respectively, versus 0.900±0.023, 1.740±0.032, 1.230±0.025 of the negative control cells, with significant differences ( t=10.79, 12.00, 7.56, P<0.05). ② Results of Transwell assay showed that the invasion numbers of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 186±12, 155±7, 75±9, respectively, versus 505±1, 245±8, 300±15 of the negative control cells, showing significant differences ( t=955.90, 163.40, 530.90, P<0.05). (4) Effects of LncRNA KCNQ1OT1 gene knockdown on PI3K/p-AKT signaling pathways. Resluts of Western blot showed that the relative repression levels of PI3K in HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.447±0.009, 0.430±0.012, 0.354±0.006, respectively, versus 0.820±0.017, 0.850±0.012, 0.531±0.001 of the negative control cells, showing significant differences ( t=18.94, 25.72, 27.46, P<0.05). The relative repression levels of p-AKT in HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.343±0.015, 0.410±0.012, 0.579±0.006, respectively, versus 0.546±0.012, 0.620±0.012, 0.830±0.012 of the negative control cells, showing significant differences ( t=10.78, 12.86, 19.02, P<0.05). Conclusions:LncRNA KCNQ1OT1 plays an important role in the occurrence and development of HCC. LncRNA KCNQ1OT1 gene knockdown can inhibit the PI3K/AKT signaling pathways, so it can significantly inhibit the proliferation, migration and invasion of HCC cells.
2. Bone marrow mesenchymal stem cells interactions with hepatocytes and hepatic stellate cells on different stiff substrates
Xiumei CAO ; Qiping HUANG ; Shuaishuai CHEN
Chinese Journal of Hepatology 2019;27(6):424-429
Objective:
To explore the effect of substrate mechanical microenvironment and cell-cell interaction on differentiation of bone marrow mesenchymal stem cells (BMSCs), intrahepatic cellular function and phenotype.
Methods:
Bone marrow mesenchymal stem cells (BMSCs)-hepatocytes (HCs) and BMSCs-hepatic stellate cells (HSCs) were co-cultured on polyvinyl alcohol (PVA) hydrogel substrates at different stiffness (4.50 ± 0.47 kPa, 19.00 ± 3.51 kPa and 37.00 ± 2.09 kPa) by non-contact co-culture method. Furthermore, the effect of substrate mechanical microenvironment on BMSCs, HCs and HSCs and the activation and proliferation of HCs under different co-cultured condition was studied. A Student's t-test was used to compare the two groups.
Results:
The expression ofα-smooth muscle actin (α-SMA) and collagenα1- I (Col1A1) in BMSCs and HSCs cultured on its own increased with increase of substrate stiffness. After 72 h, the expression of albumin (ALB) of HCs on three stiff substrates was significantly higher than that of 24 and 48 h. Moreover, the expression of ALB of HCs increased with the increase of substrate stiffness. During the co-culture of BMSCs and HSCs, BMSCs of all three stiffness substrates promoted the expression ofα-SMA, Col1A1 in HSCs, but reduced the expression of PPARγin HSCs cells, thererby promoted the activation of HSCs, with apparent stiffness at 37 kPa. HSCs promoted the expression of ABL in BMSCs at three stiff substrates, but inhibited the expression of alpha-SMA and Col1A1 in BMSCs at 37 kPa, suggesting that co-culture had inhibited the differentiation of BMSCs myofibroblasts, and promoted the differentiation of hepatocyte-like cells, especially at high stiff substrates. In the co-culture of BMSCs and hepatic parenchymal cells, BMSCs had promoted the proliferation of hepatic parenchymal cells at 4.5 kPa. Further, hepatic parenchymal cells had inhibited the expression ofα-SMA in BMSCs, and promoted the expression of Alb, with inhibition of BMSCs differentiation towards myofibroblasts.
Conclusion
The differentiation of BMSCs affects the substrate mechanical microenvironment, co-culture of HCs and HSCs. Simultaneously, affecting the function of hepatocytes in relation to the mechanical state of the substrates.
3.Analysis of clinicopathological features and prognosis of 87 low grade glioma
Jun QIU ; Liangbo GUO ; Feng GAO ; Xinping ZHOU ; Peng HUI ; Qiping HUANG ; Tao FAN
Chinese Journal of Postgraduates of Medicine 2017;40(3):204-207
Objective To investigate the clinicopathological features and prognostic factors in patients with low grade glioma. Methods Eighty- seven patients with low grade glioma confirmed by cytological examination were retrospectively analyzed. Results Among 87 patients, male accounted for 62.1%(54/87), and the average age was 36.2 years. The average followed- up time was 51.4 months (3-135 months), and 5 cases were lost in follow-up. The follow- up rate was 94.3% (82/87). The 3- year survival rate was 84.1%, and the 5- year survival rate was 66.7%. The 3- year survival rate was related to clinical symptoms, maximum diameter of tumor and the extent of resection (P<0.01 or<0.05);and the 5- year survival rate was related to years, Karnofsky score, clinical symptoms, maximum diameter of tumor, the pathology classification and extent of resection (P<0.05 or<0.01). Conclusions Low grade glioma is more common in male patients and patients under the age of 40 years. Age below 40 years, Karnofsky score ≥ 70 scores, only epilepsy symptom, tumor diameter below 6.0 cm, oligodendrogliomas, complete resection or subtotal resection are positive prognostic factors. Postoperative radiotherapy can reduce the rate of tumor progression.
4.Effect of left ventricular hypertrophy and deformation on cardiac function in patients with uremia cardiomyopathy by using quantitative speckle tracking technology
Yi ZHENG ; Xiaohong YANG ; Qiping ZHOU ; Yaoyao HUANG ; Fan ZHANG ; Liangyu WANG
Journal of Central South University(Medical Sciences) 2017;42(4):400-405
Objective:To evaluate the effect of left ventricular hypertrophy and deformation on cardiac function in patients with uremic cardiomyopathy (UCM) by using the technology of two dimensional speckle tracking imaging (2D-STI).Methods:A total of 67 UCM patients were randomly divided into the normal cardiac function group (subgroup A,32 cases) and the abnormal cardiac function group (subgroup B,35 cases)according to the New York Heart Association points (NYHA-P).A total of 30 healthy subjetcs served as the control group.Parameters including left ventricular ejection fraction (LVEF),left ventricular mass index (LVMI),left ventricular spherical index (LVSI),left ventricular myocardial mean radial strain (MRS),mean radial strain rate (MRSR),mean longitudinal strain (MLS),local systolic twist angle (STA),and mitral annulus maximum displacement (TMAD) were detected.Results:MLS,MRS,MRSR,LVSI,STA and TMAD in the Group A and Group B were lower than that in the control group (P<0.05),and LVMI in the Group A and Group B was increased than those in the control group (P<0.05);LVEE MLS,MRS,MRSR,LVSI and STAin the Group B was decreased than that in the Group A (P<0.05).MLS in the Group A and B were positively correlated with LVEF and LVSI,but negatively correlated with LVMI.Using the point of 14.10% for MLS to evaluate UCM patients with NYHA-P>4 points,the sensitivity,the specificity and Yuedden index were 90.5%,71% and 0.585,respectively.STA in UCM patients were lower than that in the control (P<0.05).Conclusion:2D-STI possesses a unique advantage in detecting left ventricular strain and strain rate on left ventricular regional function in UCM with left ventricular hypertrophy and ventricular deformation.There is no direct correlation between the left ventricular hypertrophy and ventricular deformation,but the ventricular hypertrophy and deformation are correlated with regional cardiac function and clinical cardiac function.Left ventricular regional dysfunction may occur before cardiac hypertrophy and deformation.
5.Narrative medical enlightenment on medical humanities quality education in China
Qiong YANG ; Xinhua ZHANG ; Qiping HUANG ; Gebo WEN ; Zhihan TANG ; Juan PENG
Chinese Journal of Medical Education Research 2016;15(9):943-946
Narrative medicine has brought new ideas to medical humanities education, and its con-notation is suitable to Undergraduate Medical Education Standards-Clinical Medicine (Trial). By analyz-ing the proposal and development of narrative medicine, its foundation and core, its programs and methods, as well as its forms and values, the author deepened the understanding of narrative medicine. And through the following four pathways including strengthening the renewal of ideas and concepts on medical humanities quality education, promoting the diversification of teaching ways and methods of medical humanities educa-tion, enhancing the effect of early exposure to clinical medical students, opening the path of integration narrative medicine and evidence-based medicine, the author analyzed narrative medical enlightenment on medical humanities quality education in China.
6.The effect of ziprasidone and risperidone on schizophrenia patients and the change of leptin ,adiponectin levels
Ao ZHAO ; Qiping LI ; Huiteng LI ; Chunyang LI ; Zaiping HUANG ; Huanyu XU ; Jieping GUAN
International Journal of Laboratory Medicine 2016;37(4):506-507,510
Objective To observe the effects of ziprasidone and risperidone on schizophrenia patients and the change of serum leptin and adiponectin levels .Methods Totally 80 cases of schizophrenia patients were randomly divided into ziprasidone group and risperidone group ,which were treated for 8 weeks .Measure the positive and negative symptoms scale (PANSS) score and body weight of that number ,leptin and adiponectin at baseline ,treatment 4 weeks and 8 weeks respectively for patients ,at the end of the experiment ,the results for statistical analysis .Results Two groups of 4 ,8 weeks after treatment scores compared with baseline scores dropped significantly ,the difference was statistically significant(P<0 .05) .Risperidone group after treatment ,leptin levels significantly increased body mass index ,and adiponectin levels significantly decreased ,compared with the baseline before treatment was statistically significant difference(P<0 .05) .Conclusion Ziprasidone and risperidone in treatment of schizophrenia have similar efficacy .Ziprasidone has no significant effect on body weight ,leptin and adiponectin levels in treatment of schizophrenia patients . However ,risperidone has a significant effect ,long-term use should pay attention to the side effects .
7.TLC identification and determination of total flavonoids in Compound caulis Polygoni multiflori mixture
Qiping ZENG ; Yixiang CAO ; Lishan HUANG ; Rong ZHANG
Journal of Pharmaceutical Practice 2016;34(5):424-427
Objective To establish the quality control standards for Compound caulis Polygoni multiflori mixture .Meth-ods Caulis polygoni multiflori ,cortex alibiziae and schisand chinensis were identified by TLC ,total flavonoids were deter-mined by UV .Results Caulis polygoni multiflori ,cortex alibiziae and schisand chinensis could be detected by TLC .A better linear relationship between concentration and absorbance in the range of 9 .12~27 .36 μg/ml .The linear regression equation was A=0 .034 55 C-0 .110 34 (r=0 .999 2) ,The average recovery of emodin was 101 .0% (RSD=1 .58% ) .Conclusion The method is simple ,quick ,accurate and sensitive ,which could be used as a quantitative analysis method for this preparation .
8.Observations on the Efficacy of Electroacupuncture plus a Multi-function Cervical Rehabilitation System in Treating Cervical Spondylosis
Zheng HUANG ; Qiping DAI ; Jian SUN ; Yunpeng WU ; Tao CHE
Shanghai Journal of Acupuncture and Moxibustion 2015;(11):1088-1090
Objective To investigate the clinical efficacy of electroacupuncture plus a multi-function cervical rehabilitation system in treating cervical spondylosis.Methods Sixty patients with cervical spondylosis were randomly allocated to treatment and control groups, 30 cases each. The treatment group received electroacupuncture plus treatment with a multi-function cervical rehabilitation system and the control group, electroacupuncture alone. After 30 days of treatment, changes in the pain score, the neck disability index, and cervical active range of motion and isometric muscle strength were observed in the two groups and the clinical therapeutic effects were compared between the two groups.Results The total efficacy rate was 96.7% in the treatment group and 93.3% in the control group; there was no statistically significant difference between the two groups (P>0.05). There were statistically significant pre-/post-treatment differences in the pain score, the neck disability index, and cervical active range of motion and isometric muscle strength in the two groups (P<0.01,P<0.05). There were statistically significant post-treatment differences in the neck disability index, and cervical active range of motion and isometric muscle strength between the treatment and control groups (P<0.05).Conclusions Electroacupuncture plus a multi-function cervical rehabilitation system is an effective way to treat cervical spondylosis. It can relieve the clinical symptoms in the patients.
9.Comparative study of curative effect and blood glucose and lipid in schizophrenic patients who treated with ziprasidone and risperidone
Qiping LI ; Ao ZHAO ; Huiteng LI ; Chunyang LI ; Zaiping HUANG ; Huanyu XU ; Jieping GUAN ; Tanling XIAO ; Jinhua ZHANG ; Zhanhong FU
International Journal of Laboratory Medicine 2015;(17):2471-2472,2475
Objective To observe the effects of ziprasidone and risperidone on patients with schizophrenia and their influence on bloodglucoseandlipids.Methods 96patientswithschizophrenicenrolledinthestudywererandomlydividedintotwogroups,zi‐prasidone and risperidone group ,and both were treated for 8 weeks .Their blood glucose ,blood lipid of base line and at the end of the 4th ,8th week were determined respectively .Results The positive and negative symptoms scores of the two groups by using Positive and Negative Syndrome Scale(PANSS) before and after treatment were not statistically different(P> 0 .05) .Compared with the baseline scores ,scores at the end of 4th and 8th week in both ziprasidone and risperidone groups significantly decreased(P<0 .05) ,but there was no statistically significant difference between the two groups(P>0 .05) .After 8 weeks′ treatment ,the ef‐fective rate was 91 .7% in ziprasidone groups and 89 .6% in risperidone group .There were no significant differences between the two groups(P>0 .05) .The blood lipids and glucose levels were less increased after ziprasidone treatment ,but was not statistically significant(P>0 .05) .The blood lipids and glucose levels significantly increased after risperidone treatment(P<0 .05) .Conclusion Ziprasidone and risperidone had the same effect on schizophrenia .Ziprasidone had no effect on blood glucose and lipids in schizo‐phrenic patients ,while risperidone could increase blood glucose and lipids level ,we should pay attention to the side effects of long‐term use .
10.Preparation and evaluation of insulin thiolated hyaluronic acid nanoparticles in vitro
Aiwen HUANG ; Jiali ZHAO ; Zhihong LIU ; Qiping ZENG ; Hongtao SONG
Chinese Journal of Biochemical Pharmaceutics 2014;37(7):77-80
Objective To prepare insulin thiolated hyaluronic acid nanoparticles (Ins-HA-Cys-NPs)and study its physicochemical properties. Methods The Ins-HA-Cys-NPs was prepared by ultrasonic emulsifying method,and the properties of nanoparticles including morphology,mean diameter,Zeta potential,entrapment efficiency and drug loading efficiency were studied,as well as the cryoprotectant selection.Results The prepared nanoparticles was round in appearance and the mean diameter was(178.5 ±0.8)nm,the polydispersity index was (0.214 ±0.013)and the Zeta potential was -(38.47 ±0.46 )mV,while the entrapment efficiency was (48.85 ±0.66 )%,drug loading efficiency was (4.79 ±0.13 )%;10%mannitol as cryoprotectant provided uniform and well dispersed suspension of nanoparticles with blue opalescence after redispersion.Conclusion The thiolated hyaluronic acid nanoparticles may be used as the carrier for oral drug delivery system of insulin,and it provides a basis for studies on rats in vivo.

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