1.The investigation of effect of trichostatin A to breast cancer MDA-MB-231 cell line
Journal of Chongqing Medical University 1986;0(03):-
Objective:To investigate the influence of histone deacetylase inhibitor(Trichostatin A)to multiplication,invasion, cell cycle of breast cancer MDA-MB-231 cell line.Methods:After 6-48hours treatment with the concentration(100~400nmol/L)of Trichostatin,the growth activity of MDA-MB-231 cell line was detected by MTT.The expression of ER?,MMP-9 mRNA was detected by RT-PCR with different concentration(100~400nmal/L TSA)during 24 hours to MDA-MB-231 cell line.The expression of ER?,MMP-9 protein was examined by immunohistochemistry with different concentration(100~400nmol/L TSA)during 24 hours treatment to MDA-MB-231 cell line.With cell invasion experiment to detect the change of invasive power by using different concentration(100~400nmol/L TSA)during 24 hours to MDA-MB-231 cell line.Results: Trichostatin can inhibit the growth of MDA-MB-231 cell line,which was time and dose dependent.Trichostatin can delay the cell cycle G_2/M stage,stop the cell stage in G_2 stage.Trichostatin can up-regulate the expression of ER?mRNA, down-regulate the expression of MMP-9 mRNA in cells.The invasion experiment indicate that Trichostatin can conspicuous inhibit the invasion of MDA-MB-231 cell line.Conclusions:Trichostatin can inhibit the accrementition,invasion and metastasis of MDA-MB-231 cell line evidently,and one of the mechanism may be the up-regulation of ER?and the down-regulation of MMP-9.
2.Expression and relationship between hepaCAM protein and multidrug resistance protein in renal carcinoma
Xiaoliang JIANG ; Xu LUO ; Ruming LIU
Journal of International Oncology 2016;43(8):584-587
Objective To investigate the expression and relationship between hepatocyte cell adhesion molecule (hepaCAM) protein and some multidrug resistance proteins in renal carcinoma tissue.Methods Expressions of hepaCAM,multidrug resistance associated protein (MRP),P-glycoprotein (P-gp),lung resistance protein (LRP),and topoisomerase Ⅱ (TOPO Ⅱ) protein were detected by immunohistochemistry in different areas of human renal cell carcinoma tissues and their relationships were analyzed.Results In the peripheral zone of renal tumor,hepaCAM,MRP,P-gp and LRP protein were showed positive expression.In the central region of the renal tumor,the expressions of hepaCAM,P-gp and LRP were negative or weakly positive,while the expressions of MRP and TOPO Ⅱ protein were positive.The expressions of MRP and TOPO Ⅱ protein in the central region of tumor were stronger than those in the peripheral zone of tunor (31.23 ±5.67 vs.23.89 ±4.56;45.66 ±2.34 vs.5.23 ±0.66),with statistically significant differences (t =-6.20,P =0.00;t =-100.16,P =0.00).While the expressions of other proteins (hepaCAM,P-gp and LRP) in the central region of tumor were weaker than those in the peripheral zone of tumor (3.21 ±1.12 vs.27.25±2.23;2.34±0.33 vs.51.23±3.45;4.22±1.78 vs.44.23 ± 1.45),with statistically significant differences (t =60.87,P =0.00;t =90.35,P =0.00;t =107.18,P =0.00).Correlation analysis showed that the expression of hepaCAM protein in the central region of renal carcinoma was related with the expression of MRP protein (r =0.94,P =0.01),but it was not related with the expressions of P-gp,LRP and TOPO Ⅱ protein (r=0.22,P=0.44;r=0.14,P=0.80;r=0.34,P=0.07).Conclusion The expression of hepaCAM protein in renal carcinoma may be related to tumor drug-resistance.
3.Design of automatic cap opening device for specimen container in the specimen pretreatment system of biochemistry analyzer
Xiaoliang BAI ; Shi CUI ; Dawei JIANG
China Medical Equipment 2017;14(2):1-4
Objective:To design a new automatic cap opening device consists of holding mechanism, open mechanism, improve mechanism, rotating mechanism and recycling mechanism in order to resolve the poor adaptability, complex structure and lower liability problem for specimen container in the biological specimen pretreatment system.Methods: This paper designed a automatic equipment to remove the rubber cap and screw cap. This equipment is compatible with the different specification specimen containers and the container cap, and the specimen container cap was stepped up and rotated with same power component.Results: The application of equipment has reduced the manufacturing cost and maintenance cost for specimen container, improved the system reliability, solved the current technical problems of the equipment, such as poor adaptability and lower liability. Conclusion: The design of equipment mainly adapts to CS-6400 series of automatic biochemical analyzer, and it can improve the detection efficiency of biological specimen, reduce the cross contamination and satisfy the practice necessity for clinical detection.
4.Active vitamin D reduces macrophage infiltration by TREM-1 in renal tissue of diabetic nephropathy rats
Yu ZHAO ; Yinfeng GUO ; Yuteng JIANG ; Bicheng LIU ; Xiaoliang ZHANG
Chinese Journal of Nephrology 2017;33(3):204-212
Objective To investigate the effects of active vitamin D (VD) on the expression of triggering receptor expressed on myeloid cells-1 (TREM-1) in renal tissue of diabetic nephropathies (DN) rats and to explore the impact of TREM-1 on adhesion and migration capacity of macrophage.Methods DN rat models were established by streptozotocin.Rats were randomly distributed into four groups:control (NC) group,VD group,DN group and DN+VD group (DN rats with 0.1 μg · kg-1 · d-1 calcitriol by garages).Rats were sacrificed respectively at 8 weeks and 12 weeks after treatment.Pathological changes in kidney tissue were detected and the expressions of CD68 and TREM-1 were acquired by immunohistochemistry stain and Western blotting.In vitro,RAW264.7 cells were divided into NC group,VD group,high glucose (HG) group and HG+VD group.In HG+VD group rats were treated by high glucose with 10-8 mol/L 1,25(OH)2D3.TREM-1 expression was measured by immunohistochemistry stain and Western blotting,and the ability of macrophage in migration and adhesion was evaluated by Transwell migration assay and adhesion assay.TREM-1 siRNA was transferred to silence TREM-1 expression,while plasmid of TREM-1 was transferred for high expression.Their ability of adhesion and migration in macrophage and the effect of 1,25(OH)2D3 were examined.Results (1) Compared with the NC group,the expressions of CD68 and TREM-1 were increased in DN group (P < 0.05),whereas markedly decreased in DN+VD group (P < 0.05).(2) The number of adhesion and migration cells,and the expression of TREM-1 protein in macrophage were obviously increased in HG group as compared with those in NC group (all P < 0.05);whereas above changes were markedly decreased in HG+VD group than those in HG group (P < 0.05).(3) The number of adhesion and migrated macrophage was reduced after TREM-1 siRNA intervention (all P < 0.05).VD could significantly decrease the effect of high glucose on adhesion and migrated macrophages after TREM-1 siRNA (all P < 0.05).(4) Adhesion and migration of macrophage were increased via TREM-1 overexpression (all P < 0.05),but the effects of VD on high glucose-induced adhesion and migration of macrophage were disappeared.Conclusions VD can suppress the adhesion and migration of macrophage via reducing the expression of TREM-1,and inhibit infiltration of macrophage in renal tissue of DN rats.
5.Combination treatment of lfexible/lfexible sheath and rigid ureteroscopic lithotripsy for upper and middle ureteral stones
Jian YANG ; Hongyi JIANG ; Fajun FU ; Xiaowang XIAO ; Xiaoliang GUO
China Journal of Endoscopy 2017;23(1):25-28
Objective To study the combination treatment of lfexible/lfexible sheath and rigid ureteroscopic lithotripsy (F-ul) for upper and middle ureteral stones. Methods The clinical data of patients diagnosed of upper and middle ureteral stones were collected. The treated group (110 cases):ifrstly treated with rigid ureteroscopic lithotripsy to broke and removed stones through lfexible sheath, then the lfexible ureteroscopic lithotripsy was used to broke and removed stones through lfexible sheath;The control group (110 cases):traditional operation for ureter calculi. The clinical data was compared between the two groups. Results The effective ratio of treatment group is 90.0%, which was better than that of control group (87.3%) (P>0.05). The operation time, stone processing time of treatment group were signiifcantly shorter than those of control group (P<0.05), and F-ul using time was signiifcantly much more (P<0.05). The hospitalization time and complication rate were no signiifcantly difference between the two groups (P> 0.05). Conclusion The method of combining flexible/flexible sheath and rigid ureteroscopic lithotripsy for upper and middle ureteral stones was better than that of traditional operation, which worth to be popularize in clincal treatment.
6.Preparation of calcium titanate coating on titanium and its biosafety
Chunfeng WANG ; Min SUN ; Zhanao WU ; Xiaoliang WU ; Tao JIANG
Journal of Medical Postgraduates 2015;(5):480-485
requirement of medical materials for hemolysis experiment (<5%) .MTT assay showed that , after 4 days of culture , the optical densi-ties were 0.498 ±0.218 and 0.566 ±0.266 in the 120℃12 h and 24 h groups and 0.668 ±0.268 and 0.769 ±0.213 in the 150℃12 h and 24 h groups, while after 8 days, the optical densities were 0.767 ±0.267 and 0.836 ±0.236 in the 120℃12 h and 24 h groups and 0.765 ±0.265 and 0.903 ±0.303 in the 150℃12 h and 24 h groups, all significantly higher than in the non-CaTiO3 group at 4 (0.341 ±0.143) and 8 days (0.731 ±0.121) (P<0.05). Conclusion The CaTiO3 coating on titanium is neither mutagenic nor hemolytic and has no toxicity on osteoblasts .Instead, it can promote the proliferation of osteoblasts , and therefore is a valuable coating material for implants .
7.Correlation of C - erbB - 2 Status and Histologic Features in 1 6 3 Invasive Breast Carcinomas of baoji area
Yuan CAI ; Hongying LI ; Kui JIANG ; Xiaoliang CHE ; Jianli FENG ; Su ZHENG
Chinese Journal of Primary Medicine and Pharmacy 2011;18(15):2028-2030
ObjectiveTo study the relationship between C-erbB-2 and estrogen (ER) and progesterone (PR) receptors, and the relationship between C-erbB-2, ER, PR with histologic grade. MethodsTo detect ER, PR and C-erbB-2 states by using immunohistochemical analysis and fluorescence in situ hybridization for C-erbB-2 in 163 unselected invasive breast carcinomas. ResultsC-erbB-2, ER ,PR were expressed in 21.5% ,64.4% ,44.2% of 163 cases respectivly . 5 pure mucinous carcinomas , 3 tubular carcinomas and 1 micropapillary carcinoma were ER + ( 100.0% ) 、C-erbB-2 - ( 100.0% ) and PR + (40.0% ,66.7%, 100.0% ). C-erbB-2 was positive in 22.3% of grade Ⅱ and 27.0% of grade Ⅲ invasive ductal carcinomas and negative in all grade Ⅰ invasive ductal carcinomas.ER and PR expression were decreased significantly in C-erbB-2 + tumors compared with C-erbB-2 - tumors( ER,25. 7% vs 75.0% ; PR,25.7% vs 49.2% ). Although ER or PR expression is decreased in C-erbB-2 + tumors, a substantial proportion of them still express ER or PR. ConclusionC-erbB-2 overexpression or amplifcation was limited to a minority of invasive breast carcinomas. Tumour grade was an independent predictor for ER expression. ER was expressed in small number of high-grade and in large number of grade Ⅰ invasive ductal carcinomas. C-erbB-2 overexpression or amplification essentially was limited to grades Ⅱ and Ⅲ ductal carcinomas and correlated inversely with ER or PR expression.
9.The study on the mechanism of platelet in the development of salt-sensitive hypertension via inflammation
Hongya MAO ; Yunpeng LIU ; Zihao WANG ; Xiaoliang JIANG ; Xing LIU ; Zhiwei YANG
Chinese Journal of Comparative Medicine 2017;27(5):23-30
Objective To investigate the role and mechanism of platelet in the development of salt-sensitive hypertension.Methods 25 Dahl salt-sensitive rats (Dahl SS) were divided into three groups: low-salt diet (0.12% NaCl, LS), high-salt diet (8%NaCl, HS) and high-salt diet + platelet inhibitor (8%NaCl+busulfan, HS+bus).Blood pressures were measured by tail-cuff method.After six weeks, animals were sacrificed.Platelet p-selectin expression, platelet cytosolic Ca2+ concentration, platelet-leukocyte aggregation (PLA) in peripheral blood, and immune cells infiltrated on aortic walls were assessed by flow cytometry, and serum IL-6 level was tested by ELISA in vivo.Platelets purified from SD rats were treated with normal salt (0.9%NaCl) and high salt (1.3%NaCl), then the cytosolic Ca2+ concentration and p-selectin expression of platelet were detected.Results We found that Dahl SS rats with high-salt diet, relative to low-salt diet, presented with high blood pressure and increased the ratio of platelet p-selectin expression, Ca2+ concentration.IL-6 level and PLA in peripheral blood, and the number of infiltrated immune cells on aortic walls were also significantly elevated in high-salt diet group.The whole events were ameliorated by the platelet inhibitor busulfan.Cytosolic Ca2+ concentration and p-selectin expression were also increased in purified platelets treated with high salt than those treated with low salt (P < 0.05).Conclusions Our findings suggest that high salt induced platelet activation with increased Ca2+ concentration may play an important role in the development of salt-sensitive hypertension via vascular inflammation.However, the detailed mechanisms of platelet activation and development of high blood pressure via inflammation induced by high salt intake remain to be determined.
10.Effect of macrophages on podocytes apoptosis in diabetic nephropathy
Yinfeng GUO ; Yu ZHAO ; Yuteng JIANG ; Xiaodong ZHU ; Bicheng LIU ; Xiaoliang ZHANG
Chinese Journal of Endocrinology and Metabolism 2017;33(8):680-686
Objective To investigate the effect of macrophages on podocytes apoptosis in diabetic nephropathy. Methods Differentiated mouse macrophages (RAW264.7) were exposed to normal glucose, high glucose, then the conditioned media (CM) was collected and considered as NC-CM or HG-CM, respectively. Western blotting and immunofluorescent staining were used to detect the specific markers for M1 macrophages (iNOS) and M2 macrophages (MR). ELISA was used to detect the concentration of TNF-α in the CM. Then normal PRMI 1640 media (control), NC-CM or HG-CM was added to podocytes. In some experiments, ROS inhibitors (Tempo), p38 MAPK inhibitor (SB203580), anti-TNF-α neutralizing antibody, and IgG1 isotype control were respectively added to cells with HG-CM. Besides, recombinant mouse TNF-α alone was applied to incubate podocytes. Podocytes apoptosis was accessed by Annexin V-FITC/PI and Hoechst33342 staining. DCFH-DA staining was used to analyse ROS level. Western blotting was used to detect cleaved casepase-3, p38MAPK, and p-p38MAPK protein. Results Macrophages were activated when exposed to high glucose, displaying pro-inflammatory M1 polarization with higher iNOS and lower MR expression. HG-CM but not NC-CM trigged podocytes apoptosis, up-regulated ROS, cleaved casepase-3 and p-p38MAPK. However, the podocytes apoptosis trigged by HG-CM was abolished by either a ROS inhibitor (Tempo) or a p38 MAPK inhibitor (SB203580). Additionally, TNF-α was increased in the HG-CM. TNF-α protein in macrophage was aslo increased when exposed to high glucose. Anti-TNF-α neutralizing antibody blunted the apoptotic response, excess ROS generation and p-p38 MPAK expression in podocytes induced by HG-CM. Moreover, addition of recombinant TNF-α similarly led to podocytes apoptosis, increased ROS and p38 MPAK expression. Conclusion M1 macrophages activated by high glucose released TNF-α to promote podocytes apoptosis via ROS-p38 MAPK pathway.