1.Effect of exogenous small RNA transfection on the growth of renal clear cell carcinoma cells
Yong WANG ; Yonglian GUO ; Lin CHEN ; Guohao LI ; Jiajun YU ; Wei CHENG
The Journal of Practical Medicine 2017;33(10):1572-1575
Objective To investigate the effect of exogenous small RNA (dsP21-397) transfection on growth of human renal clear cell carcinoma cell lines A-498 and Caki-1. Methods The dsControl(control group) and dsP21-397(experimental group)were transfected into A-498 and Caki-1,respectively. The expression of p21 mRNA was analyzed by qRT-PCR. The expression of p21 protein,CDK4 and Cyclin D1 proteins were detected by Western blot. The cell cycle distribution was examined by flow cytometry(FCM). MTS assay and colony formation assay were used to analyze cell viability and proliferation ability. Results Compared with dsControl,p21 mRNA levels in A-498 and Caki-1 cells increased to 2.55-fold(P<0.01)and 2.18-fold(P<0.01),respectively,after transfection with dsP21-397. The expression of p21 protein was up-regulated while the expression of CDK4 and CyclinD1 were down-regulated. The percentage of cells in G0/G1 phase significantly increased after transfection of dsP21-397,and the proportion of cells in S phase and G2/M phase significantly decreased. The activity of A-498 and Caki-1 cells significantly decreased and the number of colonies in the dsP21-397 group was significantly lower. Conclusions dsP21-397 can significantly activate p21 protein expression,down-regulate the cell cycle-associated proteins,and inhibit the growth of renal clear cell carcinoma cells.
2.Analysis of Clinical Manifestations of Dengue Fever in Guangzhou Higher Education Mega Center in 2014
Shijun GUO ; Hairong CAI ; Yonglian HUANG ; Juming HUANG ; La ZHANG ; Ying CHEN ; Zhongde ZHANG ; Bojun CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(5):796-798,807
Objective To analyze the clinical characteristics of dengue fever (DF) in Guangzhou Higher Education Mega Center (HEMC) in the year of 2014. Methods A retrospective analysis was carried out in the clinical data of 487 cases of DF patients. Results The incidence of DF accounted for 27.18% ( 527/1939) of the total emergeney fever cases. In 487 DF cases with detailed data, 261 were male, and 226 were female; 312 were young aged, 88 were middle aged, and 87 were old aged. For the systemic symptoms, fever accounted for 87.5%, headache 48.3%, and muscular soreness 47.6%. Cough ( 16.6%) , pharynx pain ( 16.8%) and running nose ( 10.9%) were common in respiratory symptoms; poor appetite ( 14.4%) and nausea ( 10.3%) were common in gastrointestinal symptoms. The abnormal laboratory parameters were mainly shown as WBC count ( 48.25%) and PLT count ( 41.68%) , following by creatine kinase ( CK, 39.49%) , aspartate aminotransferase ( AST, 34.12%) , and lactate dehydrogenase ( LDH, 31.96%) . Less cases had abnormal renal function. The distribution of Chinese medical syndrome types was shown as damp-heat blockage (65.7%), syndrome involving Weifen and Qifen simultaneously (23.6%), and Qi-yin deficiency (10.7%). Conclusion In the year of 2014, DF in Guangzhou HEMC occurred mainly among the youth people, the incidence of male DF was similar to that of the female DF, and DF cases usually have the primary symptoms of fever, aversion to cold, headache and muscular soreness. Bleeding is seldom seen in the DF patients, a few cases are complicated with the damage in the blood, liver and myocardium, and most of them have good prognosis.
3. Curative effects of Ningmitai capsules on the residual fragments and postoperative complications following lithotripsy treatment of upper urinary calculi
Wei XIANG ; Yonglian GUO ; Jingyu ZHANG ; Tao ZHENG ; Chuanhua ZHANG
Chinese Journal of Surgery 2018;56(10):776-780
Objective:
To observe the clinical efficacy of Ningmitai capsules, a traditional Chinese medicine using for clearing heat and dampness, in the treatment of residual fragments and postoperative complications following lithotripsy for upper urinary stones.
Methods:
During October 2016 and March 2018, patients from Wuhan 1st Hospital, Wuhan 2nd Hospital, Wuhan 3rd Hospital, and Wuhan Puai Hospital having upper urinary residual fragments following minimally-invasive stone treatment were randomly assigned to control group and Ningmitai group with a proportion of 1∶3. The patients in control group were treated with antibiotics or sodium diclofenac suppository on demand, while patients in Ningmitai group took additional Ningmitai capsule orally (4 capsules per time, 3 times per day). The observation was started when a patient was enrolled in this study and continued for a maximum of 12 weeks or until stone-free status. During the observation, the stone expulsion time, stone-free time, stone-free rate were observed, and the difference in curative effect between the two groups on postoperative complications such as pain and infection were compared. Statistical analysis was done using
4.Effect of microRNA-1180 transfection on growth of renal cell carcinoma
Yong WANG ; Yonglian GUO ; Lin CHEN ; Guohao LI ; Jiajun YU ; Wei CHENG
Chongqing Medicine 2018;47(6):736-739
Objective To investigate the effect of microRNA-1180 transfection on the growth of renal cell carcinoma lines 786-O and ACHN.Methods The renal carcinoma cells were divided into the two groups:control group (transfecting dsControl) and experimental group (transfecting miR-1180).The expression change of p21 mRNA was detected by qRT-PCR.Western blot was conducted to analyze the expression changes of p21,CDK4,CDK6 and CyclinD1 proteins.Flow cytometry (FCM) was used to detect the cell cycle change.The MTS assay was conducted to detect the cell viability and the colony forming assay was performed to examine the cell proliferation ability.Results The qRT-PCR results showed that compared with the negative control dsControl group,after miR-1180 transfection,the expression level of·p21 mRNA in 786-O and ACHN cells was up-regulated to 2.54-fold and 2.49-fold respectively(P<0.01).The expression trend of p21 protein was consistent with qRT-PCR results.The expression of CDK4,CDK6 and CyclinD1 proteins were significantly down-regulated.The FCM results showed that the proportion of cells in G0/ G1 phase was significantly increased after transfection of miR-1180,but the proportion of cells in S phase and G2/M phase was decreased significantly,indicating that the cell cycle was arrested in G0/G1 phase.The MTS assay results showed that compared with the dsControl group,the viability of the two kinds of renal carcinoma cells was significantly decreased.The colony formation assay showed that the number of colonies formed in the miR-1180 group was smaller,indicating the proliferation ability of miR-1180 transfected cells was decreased.Conclusion miR-1180 can significantly activate the p21 protein expression and inhibit the growth of renal carcinoma cell lines 786-O and ACHN.
5.miR-1180-5p inhibits proliferation, migration and invasion of prostate cancer cells by activating CDKN1Agene expression
WANG Yong ; GUO Yonglian ; CHEN Lin ; LI Guohao ; Ying Chengcheng ; CHENG Wei
Chinese Journal of Cancer Biotherapy 2018;25(7):698-703
Objective: To study the effects of microRNA-1180-5p (miR-1180-5p) on malignant biological behaviors of prostate cancer VCAP and LNCaP cells and the possible mechanisms. Methods: dsControl (dsControl group) and miR-1180-5p (miR-1180-5p group) were constructed and then transfected into two prostate cancer cell lines VCAPand LNCaP. qPCR and Western blotting were used to analyze the changes in mRNA and protein expressions of CDKN1A, Cyclin D1 and CDK6 after transfection. Cell cycle distribution, proliferation activity, clone formation capacity, cell migration and invasion ability were detected by flow cytometry, MTT assay, colony culture assay and Transwell assay, respectively. Results: qPCR results showed that compared with dsControl, CDKN1A mRNA levels in VCAP and LNCaP cells transfected with miR-1180-5p were up-regulated significantly, while the mRNA expressions of Cyclin D1 and CDK6 were significantly down-regulated (all P<0.01). Western blotting result was consistent with that of qPCR. The percentage of cells in G0/G1 phase was increased after transfection of miR-1180-5p (P<0.05), but the proportion of cells in S phase and G2/M phase was decreased and the cell cycle was arrested at G0/G1 phase (P<0.05). The proliferation activity of the two prostate cancer cells was significantly lower than that of the dsControl group after miR-1180-5p transfection (P<0.05), and the number of colonies in the miR-1180-5p group was significantly lower than that in the dsControl group (P<0.01). In the meanwhile, the cell migration and invasion ability in miR-1180-5p group was decreased (P<0.01). Conclusion: miR-1180-5p can significantly activate CDKN1A gene expression in prostate cancer cells and further inhibit the proliferation, migration and invasion of prostate cancer cells.