1.Correlation between the expression of activator protein-1 (c-Fos/c-Jun) mRNA and gingival inflammation
Dongwang ZHU ; Ling GAO ; Jiashan LI ; Shaoyun JIANG
International Journal of Biomedical Engineering 2017;40(3):197-200,215
Objective To study the correlation between the expression of activator protein-1 (c-Fos/c-Jun) mRNA and gingival inflammation,so as to discuss the pathogenesis of periodontitis.Methods The gingival tissues were divided into three groups according to the gingival index (GI),including GI=0 group (control group,14 cases),GI=1 group (15 cases) and GI=2 group (11 cases).The total RNA in each gingival tissue was extracted,and cDNA was synthesized by reverse transcription synthesis.The expressions of c-Fos and c-Jun mRNA in healthy gingival tissue (GI=0 group) were detected by reverse transcription-polymerase chain reaction.The levels of c-Fos and c-Jun mRNA in all the groups were detected by real-time quantitative PCR.Results Both c-Fos and c-Jun mRNA was expressed in healthy gingival tissues.The levels of c-Fos and c-Jun mRNA in GI=1 group was 15.58±9.19 and 3.47± 1.77,respectively,which was significantly higher than 1.31±1.03 and 1.32±0.94 in GI=0 group,and the differences were statistically significant (all P<0.05).The level of c-Fos mRNA in GI=2 group was 3.01±1.48,which was lower than that in GI=1 group (P<0.05) and higher than that in GI=0 group (P<0.05).The level of c-Jun mRNA in GI=2 group was 1.48±0.65,which was lower than that in GI=1 group,and had no significant difference with GI=0 group (P> 0.05).Conclusions Activator protein-1 (c-Fos/c-Jun) is associated with the degree of gingival inflammation,suggesting that it is involved in the occurrence and development of gingival inflammation.
2.Prognostic factors in 339 patients with T2N0M0 colorectal cancer
Bin XU ; Lin YU ; Lizhong ZHAO ; Li ZHANG ; Dongwang MA
Chinese Journal of General Surgery 2016;31(7):553-556
Objective To find T2N0M0 colorectal cancer patients at high risk for relapse or metastasis.Methods From January 1993 to December 2014,339 patients with histologically confirmed stage T2N0M0 primary colorectal cancer treated by radical surgery with complete clinical follow-up data were enrolled into this study.Survival rates were calculated using Kaplan-Meier method,and survival cures were compared using the Log-rank test.Cox proportional hazards model was used to analyze the significant factors defined in univariate test.Results The 5-year and 10-year overall survival rates were 83.0% and 68.9%,respectively.Male gender,old age,lymphovascular involverment,perineural invasion,poor differentiation and invasive micropapillary carcinoma were associated with low cancer-specific survival rates in Kaplan-Meier analysis.Multivariate analyses revealed male gender,old age,lymphovascular involverment,poor differentiation and invasive micropapillary carcinoma as significant independent factors predicting poor prognosis.Conclusions Male gender,old age,lymphovascular involvement,poor differentiation and invasive micropapillary carcinoma are risk factors predicting poor prognosis for T2N0M0 colorectal patients.
3.Study on differentiation pluripotency of human gingival fibroblasts induced in vitro
Shaoyun JIANG ; Yufei TAO ; Yang LI ; Liting SONG ; Dongwang ZHU ; Jiayin DENG
Tianjin Medical Journal 2015;(7):713-716
Objective To investigate the pluripotency of human gingival fibroblasts (hGFs), and provide a novel cell source for tissue engineering. Methods With informed consent from volunteers, fresh and healthy gingiva were collected. The hGFs were obtained from the gingiva by tissue culture. The third passage of hGFs was cultured in osteogenic medium, chondrogenic medium and adipogenic medium. Cells without differentiation were taken as control. Cells were examined by al?kaline phosphatase (ALP) staining, Alizarin red staining, Alcian blue staining and oil red O staining for detecting of the abili?ty of differentiation pluripotency. Real-time polymerase chain reaction was applied to examine the expression of osteogenic marker genes ALP, runt-related transcript factor 2 (Runx2), chondrogenic marker aggrecan (AGR) and adipogenic marker peroxisome proliferator-activated receptor gamma 2 (PPARγ2). Results The hGFs cultured in osteogenic medium showed massive violet deposit at day 7 and calcium nodulus at day 28, meanwhile, the expressions of ALP and Runx2 were higher than those of control (P<0.01). In chondrogenic group cells were found blue deposit at day 14. In adipogenic group lipid-filled droplets stained with oil red O were found in cells at day 14. However, hGFs in control group had no any positive stain?ing. Furthermore, expressions of AGR and PPARγ2 were significantly higher than those of control (P<0.01). Conclusion Human gingival fibroblasts have the pluripotency of osteogenic, adipogenic and chondrogenic differentiation.
4.Effects of stably silencing FOXM1 by shRNA on cell growth of hepatocellular carcinoma in vitro
Hongcheng SUN ; Min LI ; Jilin LU ; Dong JIN ; Dongwang YAN ; Chongzhi ZHOU ; Junwei FAN ; Huamei TANG ; Zhihai PENG
Chinese Journal of General Surgery 2011;26(5):398-401
Objective To evaluate the effect of sustained silencing Forkhead box Ml (F0XM1) gene by short-hairpin RNA (shRNA) expression vector on cell growth of hepatocelluar carcinoma (HCC) in vitro.Methods Four shRNA expression vectors targeting different sequences of human F0XM1 mRNA were constructed.The expression vector with the best interfering effect and the negative control plasmid were used to transfect HCC cell line QGY-7703, stably transfected cell clones were selected by neomycin (G418).Cell growth was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and colony formation was assessed by clonogenic assay.Cell apoptosis was detected by double staining with APC conjugated Annexin V and PI.Results F0XM1 protein was detected with different levels in all these studied human cell lines.The expression vector shRNA-1026 exhibited excellent interference effect after transient transfection, which showed 38.5% and 53.2% reduction of FOXM1 mRNA and protein level respectively.The growth of QGY-7703 cells was inhibited after stable inhibition of FOXM1 expression by shRNA-1026, which was indicated by decreased absorbance value of the test group after culture for 48, 72 and 96 h compared to control group (t = 10.830,3.578 and 5.734 respectively, P < 0.05).Stable inhibition of F0XM1 also led to reduced colony formation ( t = 5.336, P < 0.05 ) and increased apoptosis of QGY-7703 cells in comparison to control cells (t = 6.827, P < 0.05 ).Conclusions Stable silencing F0XM1 gene by shRNA suppresses the growth of HCC cells in vitro.
5.Analysis of colorectal cancer screening practices in the general population of Tianjin
Lizhong ZHAO ; Weihua ZHANG ; Dongwang MA ; Wen LI ; Yajing CAO ; Honglei WANG ; Chen XU ; Songli SHI ; Junhui HEI ; Ximo WANG
Chinese Journal of Clinical Oncology 2015;(15):760-764
Objective:Colorectal cancer screening was performed on a general population with age ranging between 40 and 74 years old to evaluate the screening effects of questionnaire survey, fecal occult blood (FOB) test, and colonoscopy, as well as to provide some implications of colorectal cancer screening strategies. Methods: Two-step screening model of questionnaire survey combined with FOB test was applied for the screening. Colonoscopy was conducted in a high-risk population identified through preliminary screening as final diagnosis. Results:Based on the 2,117,304 cases screened, the screening compliance was 39.72%, and 126,118 cases (5.96%) were identified as high risk. Colonoscopies were performed on 25,837 cases, of which 8,095, 1,236, 134, 112, and 336 were identified as adenoma, advanced adenoma, severe dysplasia lesions, early cancer, and advanced cancer, respectively. The early stage di-agnostic rate was 81.52%. Conclusion:The colorectal cancer screening method performed in Tianjin can significantly concentrate on the high-risk population with colorectal cancer, increase the positivity rate of total colonoscopy, and economize medical resources.
6.Cyclin D1 as a potential predictive biomarker for TPF induction chemotherapy in cN2 patients with oral squamous cell carcinoma
Wenwen SUN ; Xiao TANG ; Dongwang ZHU ; Jie MA ; Hailong MA ; Ying LIU ; Yiran TAN ; Yaoyao TU ; Lizhen WANG ; Jiang LI ; Laiping ZHONG ; Chenping ZHANG ; Zhiyuan ZHANG
Chinese Journal of Clinical Oncology 2015;42(16):791-795
Objective:To investigate the potential prognostic value of cyclin D1 expression in patients with locally advanced oral squamous cell carcinoma (OSCC) and its relationship with taxol (Docetaxel)/cisplatin plus 5-fluorouracil (TPF) induction chemothera-py. Methods:A total of 256 patients with locally advanced OSCC were selected from Shanghai Ninth People's Hospital of Shanghai Ji-ao Tong University School of Medicine between March 2008 and December 2010 as the objects of study in this prospective randomized clinical trial. The effect of TPF induction chemotherapy was investigated. Immunohistochemical staining against cyclin D1 was per-formed in the pretreatment biopsy specimen of the patients. The relationship between cyclin D1 expression and prognostic data of the TPF induction arm and control arm was analyzed. Results:Cyclin D1 expression was detected in 232 out of the 256 patients. Patients with low cyclin D1 expression showed significantly better overall survival (OS) (P=0.001), disease-free survival (DFS) (P=0.003), lo-coregional recurrence-free survival (LRFS) (P=0.004), and distant metastasis-free survival (DMFS) (P=0.001) than those with high cy-clin D1 expression. No significant differences existed in OS, DFS, LRFS, or DMFS between the patients with TPF induction chemother-apy and the control. Cyclin D1 expression levels were not predictive of the benefit from TPF induction chemotherapy in the overall pop-ulation. However, patients with nodal stage cN2 and high cyclin D1 expression, who were undergoing TPF chemotherapeutic regimen, showed significantly higher OS (P=0.024) and DMFS (P=0.024) than cN2 patients with high cyclin D1 expression but undergoing stan-dard surgical treatment. Conclusion:Cyclin D1 can be used as a prognostic biomarker for patients with locally advanced OSCC. Fur-thermore, cN2 OSCC patients with high cyclin D1 expression can receive long-term benefit from the addition of TPF induction chemo-therapy to standard surgical treatment.
7.Effects of total flavonoids of Bidens pilosa on insulin resistance in HepG 2 cells
Xiaojun PANG ; Linlin LU ; Dongwang LI ; Yiyu ZHAO ; Guoyong LIU
China Pharmacy 2022;33(8):968-974
OBJECTI VE To explore the effects of total flavonoids of Bidens polisa L.(TFB)on insulin resistance (IR)of HepG2 cells. METHODS B. polisa L. was refluxed and extracted with 80% ethanol to obtain TFB. Palmitic acid was used to induce IR mode of HepG 2 cells in vitro . The effects of low-concentration ,medium-concentration and high-concentration (20,40, 80 mg/L) of TFB on the consumption of glucose were investigated. Using metformin as positive control ,the effects of low-concentration,medium-concentration and high-concentration (20,40,80 mg/L)of TFB on the protein expression of insulin receptor substrate- 1(IRS-1),c-Jun N-terminal kinase (JNK)and protein kinase C (PKC)were investigated. Molecular docking technology was used to explore the interaction between eight main active components of TFB such as quercetin ,quercitrin and IRS-1,JNK and PKC proteins. RESULTS The glucose consumption of TFB low-concentration ,medium-concentration and high-concentration groups were increased significantly (P<0.05 or P<0.01). Compared with normal group ,the expression of IRS-1 and JNK protein in the model group decreased significantly ,and the expression of PKC protein increased significantly (P< 0.01). Compared with model group ,the protein expression of IRS- 1 and JNK could up-regulated while the protein expression of PKC down-regulated in TFB low-concentration ,medium-concentration and high-concentration groups and metformin positive control group (P<0.05 or P<0.01). The score of molecular docking energy between maritimetin in TFB and IRS- 1 protein was -7.9 kcal/mol(1 kcal=4.816 kJ). The scores of molecular docking energy of maritimetin ,rutin and JNK protein were -9.3 kcal/mol. The score of molecular docking energy between quercitrin and PKC protein was -4.9 kcal/mol. Interactions between components and proteins included forming hydrogen bonds ,hydrophobic bonds and so on. CONCLUSIONS TFB can significantly improve IR of HepG 2 cells,the mechanism of which may be related to the regulation of protein expression of IRS ,JNK and PKC. Maritimetin,rutin and quercitrin may be potential active ingredients for improving IR.