1.HMGB1 activates NF-κB/αvβ3 to promote A549 cells migration and invasion
Jianhua ZHU ; Na HU ; Lan ZHAO ; Yan HUANG
China Oncology 2015;(2):105-111
Background and purpose:High mobility group 1 (HMGB1), frequently found to be over-expressed in many human tumors, plays an important role in tumor progress and metastasis. This study aimed to investigate the mechanism of HMGB1 promoting A549 cell metastasis.Methods:A549 cells were untreated or treated with HMGB1 (200 ng/mL) in absence or presence of NF-κB inhibitors 6-amino-4-quinazoline (QNZ, 40 nmol/L) or Bortezomib (Bort, 20 nmol/L). Scratch assay and Transwell assay were performed to evaluate A549 cells migration and invasion ability. The activity of NF-κB was examined by luciferase reporter assay. NF-κBp65 and αvβ3 expressions were detected by Real-time RT-PCR or Western blot.Results:HMGB1 increased A549 cells migration and invasion ability. HMGB1 enhanced NF-κB protein level and NF-κB activity in A549 cells. Real-time RT-PCR and Western blot showed that HMGB1 up-regulated αvβ3 expression in A549 cells. NF-κB inhibitors QNZ or Bort reserved the promot-ing effects of HMGB1 on A549 cells migration and invasion, NF-κB expression and activity as well as αvβ3 expression. Conclusion:HMGB1 promotes A549 cell migration and invasion through activating NF-κB and up-regulating αvβ3.
2.Expression of integrin-linked kinase in human bladder transitional cell carcinoma and the related significance
Jianhua LAN ; Delin WANG ; Zongke YANG ; Zeng LI
Journal of Third Military Medical University 2003;0(13):-
Objective To investigate the roles of integrin-linked kinase (ILK) protein and mRNA in the development of human bladder transitional cell carcinoma (BTCC). Methods The expressions of ILK protein and ILK mRNA in specimens from 56 cases of human BTCC and from 30 cases of adjacent normal bladder were detected by immunohistochemistry S-P method and reverse transcription polymerase chain reaction (RT-PCR), respectively. The correlation of the expressions of ILK protein and mRNA to the clinicopathological parameters of human BTCC was analyzed statistically. Results The positive rates of ILK protein and mRNA in malignant specimens were 53.6% (30/56) and 58.9% (33/56), respectively, while those in the adjacent normal bladder tissue were 10% (3/30) and 13.3% (4/30), respectively. Significant difference was found in the expressions of ILK protein and mRNA between BTCC and adjacent normal bladder specimens (P0.05). Conclusion ILK gene transcription and protein expression may be involved in the developmental process of BTCC. ILK might be a new marker for early diagnosis of BTCC and a new target gene for BTCC treatment.
3.Isolation and initial identification of tumor stem-like cells from human prostatic carcinoma cell line PC-3
Zeng LI ; Delin WANG ; Jianhua LAN ; Zongke YANG
Medical Journal of Chinese People's Liberation Army 2001;0(11):-
Objective To isolate and identify prostate cancer stem-like cells from human prostatic carcinoma cell line PC-3,in order to set a foundation for further investigation of human prostate cancer stem cells (hPCSC). Methods Prostatic carcinoma cells line PC-3 was cultivated in serum-free medium (SFM) supplemented with growth factor,with the same cells cultivated in serum-supplemented medium (SSM) supplemented with growth factor to serve as the control. The proportion of prostate cancer stem-like cells was detected by flow cytometry,and prostate cancer stem-alike cells were identified by cell immunofluorescence,then the proliferating potentiality and doubling time with prostatic carcinoma cells PC-3 were compared by MTT. The process of inducing differentiation and the dual immunofluorescence expression after differentiation of prostate cancer stem-alike cells were observed. Results In SFM consisting of human epidermal growth factor (EGF),human basic fibroblast growth factor (bFGF) and human leukaemia inhibitory factor (LIF),a few PC-3 cells could survive and form suspended cell spheres,and the proportion of prostate cancer stem-alike cells (1.43%) was significantly increased compared with that cultured in SSM (0.41%,P
4.Effects of insulin on cardiac fibroblast proliferation and cardiac myocyte hypertrophy
Huashan HONG ; Lan LIN ; Yibo WANG ; Qiong JIANG ; Jianhua CHEN
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To study the effect of insulin on proliferation and hypertrophy of cardiac myocytes and its role in the induction of cardiac hypertrophy. METHODS: 1. The neonatal rat cardiac myocytes and cardiac fibroblasts were cultured respectively and identified with light microscopy, electron microscopy and immunocytochemistry. 2. Cell proliferation was measured with cell number, metabolic activity and DNA synthesis (with WST-1, BrdU enzyme-linked immunosorbent assay ) and the percentage of S+G 2+M in cell cycle (by flow cytometry ). 3.Cell hypertrophy was evaluated by cell protein content (Coomassie Briliant Blue's method). RESULTS: 1. The cultured cells showed the characteristic of cardiac myocytes and cardiac fibroblasts, respectively. 2. After being treated with insulin, the cell number, absorbance of BrdU incorporation and WST-1 cleavage products and the percentage of S+G 2+M of cardiac fibroblasts increased significantly ( P 0.05). 3. Protein content of cardiac myocytes increased significantly in a dose-dependent manner ( P
5.Clinical Application of Coronary Angiography with 16-slice Spiral CT
Wei FU ; Shixin CHEN ; Yanhong LAN ; Xiaohua CHEN ; Jianhua LI
Journal of Practical Radiology 2001;0(08):-
Objective To evaluate the influencing factors on image quality and the value of clinical application of coronary artery angiography with 16-slice spiral CT.Methods 16-slice spiral CT angiography was performed in 31 suspected coronary heart disease(CHD).All patients were examined in ordinary scanning (slice 3 mm) and coronary artery scanning (slice 1 mm)with the technique of retrospective electrocardiography gating,and multiplanar reconstruction(MPR),maximum intensity projection(MIP) and volume reconstruction technology(VRT) were done.The showing rate of coronary arteriae,the detecting rate of CHD and the influencing factors on image quality were evaluated by CT.Results 16-slice spiral CT could clearly show RCA1,RCA2,LAD1,LAD2,,LCX1.The image quality was affected by heart rate,breath movement and much calcification in artery,etc.Conclusion 16-slice spiral CT is of significant value in screening and diagnosing coronary artery disease.
6.Efficacy of maggot debridement therapy in the treatment of diabetic foot ulcers
Aiping WANG ; Kechun JIANG ; Ying CAI ; Jianhua YU ; Wei WANG ; Shuangwang YANG ; Lan LIU
Chinese Journal of Endocrinology and Metabolism 2011;27(6):482-484
Maggot debridement therapy(MDT)and conventional therapy were applied to diabetic foot ulcers(DFUs)with Wagner 2-4 respectively. The results showed that the prognosis was better in MDT group than in conventional therapy group. The reduction of circumference, square and volume of DFUs, healing rate of DFUs, decrease in number of bacterial species, and cost of antibiotics were all improved in the former group. The results suggest that MDT offers a fresh and effective method in the treatment of DFUs.
7.Role of ATP-sensitive potassium channels in inhibitory effect of hydrogen sulfide on high glucose-induced inflammation mediated by necroptosis in H9c2 cardiac cells
Weijie LIANG ; Meiji CHEN ; Jianhua HE ; Wenzhu ZHANG ; Fei CHENG ; Jun LAN ; Jianqiang FENG ; Huimin HUANG
Chinese Journal of Pathophysiology 2016;32(8):1364-1369
[ ABSTRACT] AIM:To investigate the role of ATP-sensitive potassium ( KATP ) channels in the inhibitory effect of hydrogen sulfide ( H2 S) on high glucose ( HG)-induced inflammation mediated by necroptosis in H 9c2 cardiac cells. METHODS:The expression levels of receptor-interacting protein 3 ( RIP3; an indicator of necroptosis ) and cyclooxyge-nase-2 (COX-2) were determined by Western blot.The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α( TNF-α) were detected by ELISA .RESULTS:After H9c2 cardiac cells were treated with 35 mmol/L glucose ( HG) for 24 h, the expression of RIP3 was significantly increased .Pre-treatment of the cells with 100 μmol/L diazoxide ( DZ; a KATP channel opener) or 400 μmol/L NaHS (a donor of H2S) for 30 min considerably blocked the up-regulation of RIP3 induced by HG.Moreover, pre-treatment of the cells with 100 μmol/L 5-hydroxydecanoic acid (5-HD; a KATP channel
blocker) attenuated the inhibitory effect of NaHS on HG-induced up-regulation of RIP3.On the other hand, co-treatment of the cells with 100μmol/L necrostatin-1 ( a specific inhibitor of necroptosis ) or pre-treatment of the cells with 100 μmol/L DZ or 400 μmol/L NaHS attenuated HG-induced inflammatory responses , evidenced by decreases in the expression of COX-2 and secretion levels of IL-1βand TNF-α.However , pre-treatment of the cells with 100 μmol/L 5-HD significantly attenuated the above anti-inflammatory effects of NaHS.CONCLUSION:KATP channels play an important role in the inhib-itory effect of H2 S on HG-induced inflammation mediated by necroptosis in H 9c2 cardiac cells.
8.Expression of CD133, CD117, and Ki-67 in human osteosarcoma and their clinical significance
Jinluan LIN ; Jianhua LIN ; Zhaoyang WU ; Wenbin LAN ; Xiang LI ; Weinan LIU ; Jinyi FENG ; Fasheng WANG
Chinese Journal of Clinical Oncology 2014;(5):305-310
Objective:This study aims to investigate the protein expression of CD133, CD117, and Ki-67 in human osteosarcoma tissues and explore their relationships with the clinico-pathological features and risk of osteosarcoma. Methods:Immunohistochemical method was used to examine the protein expression of CD133, CD117, and Ki-67 gene in the paraffin specimens of 55 and 20 cases of osteosarcoma and osteochondroma, respectively. SPSS17.0 statistical software was used to explore the relationships among the expressions of CD133, CD117, and Ki-67 gene and the biological behavior and prognosis of osteosarcoma. Test criterion:P<0.05 was considered statistically significant. Results:The positive expression rates of CD133, CD117, and Ki-67 were significantly higher in the osteosarcoma tissues than in the benign osteochondroma tissues, and the differences were significant (P=0.016, P=0.008, and P<0.001, respectively). The mean survival and metastasis time were shorter in the CD133 or Ki-67 positively expressed osteosarcoma patients than in those with CD133 or Ki-67 negatively expressed osteosarcoma. The differences were significant (P<0.05). The expression of CD133 and Ki-67, surgical staging, and distal metastasis were relevant to the prognosis of osteosarcoma patients. The expression of CD133 and distal metastasis are the independent risk factors that affect the prognosis of these patients. Conclusion:The positive expressions of CD133 and Ki-67 closely correlated with the occurrence and progression of osteosarcoma, and may be used as an indicator for prognosis of the cancer.
9.Correlation of ApoE gene polymorphism and serum uric acid in Ningxia Hui Autonomous Region
Jie WU ; Ling QIU ; Xinqi CHENG ; Qing WANG ; Lan NI ; Xiuzhi GUO ; Qian CHENG ; Jianhua HAN ; Tao XU ; Guangjin ZHU
Chinese Journal of Laboratory Medicine 2014;(6):434-438
Objective Toexplore the correlation of ApoE gene polymorphism and serum uric acid levels in Ningxia Hui Autonomous Region , China.Methods A case-control study.October 2011 to November 2011, five hundred twenty eight ( 296 male, 232 female ) apparently healthy individuals were studied.Questionnaires and physical examinations were performed by standard operation procedure.Fasting blood was collected for biochemistry testing including serum lipid parameters , uric acid concentration and creatinine levels.The multi-ARMS PCR was applied to determine ApoE genotypes ,and the relation of ApoE genotypes with serum lipid parameters and uric acid levels were analyzed.Non-normal distribution were compared using cause and inspection.Results The common six kinds of ApoE genotype can be detected.The total cholesterol ( TC) ,low density lipoprotein cholesterol ( LDL-C) and uric acid ( UA) levels in different genotype subgroups had statistical differences.The individuals with ε2/3 genotype had a significantly greater reductions in TC and LDL-C levels but increment in uric acid concentration than those withε3/3 and ε3/4 genotype (P<0.05).The effect of ApoE gene polymorphism on uric acid levels still remained significantly after adjustment for age , gender , region and other factors.Conclusion The ApoE polymorphism is associated with serum uric acid levels and individuals with ε2 allele have higher serum uric acid levels.
10.Prevalence of metabolic syndrome in population at high-risk for diabetes in Changfeng community of Shanghai
Xusheng CHEN ; Sunfang JIANG ; Jianhua SHENG ; Xuying SHEN ; Haidong KUANG ; Yuehong SHI ; Yun LEI ; Lan JIN ; Ting ZHOU ; Jing ZHU ; Xin GAO
Chinese Journal of General Practitioners 2008;7(10):674-676
Objective To investigate prevalence of metabolic syndrome (MS) in population at high-risk for diabetes in Changfeng community of Shanghai. Methods Totally, 982 persons aged 40-79 years without previous history of diabetes, but with risk factors for diabetes from Changfeng community in Shanghai were screened by questionnaire, physical examinations, lipid profile and oral glucose tolerance test. Prevalence of MS and its clinical characteristic were observed. Results Overall prevalence of MS was 44. 6% (438/982) in Changfeng community, significantly higher in women of 48.6% (341/701) than that in men of 34. 5% (97/281) (P<0.01). Prevalence of MS increased with age in women (P <0. 01 ), but not in men (P>0.05). Components of MS ranking from high to low were hypertension of 66. 5% (653/982), abdominal obesity of 62.8% (617/982), decreased high-density lipoprotein-cholesterol (HDL-C) of 49. 4% (485/982), increased triglyceride (TG) of 46. 0% (452/982) and increased fasting glucose of 35.7% (351/982). There were 55.9% (245/438) of MS patients with three or more MS components in addition to abdominal obesity. Conclusions Prevalence of MS is higher in community population at high-risk for diabetes and more than half of the MS patients have three or more risk factors for cardiovascular disease.