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.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.
3.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.
4.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 .
5.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.
6.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.
8.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.
9.The regulation of dilated cardiomyopathy by dopamine D5 receptor through inhibiting oxidative stress
Zihao WANG ; Yongyan HU ; Wenjie WANG ; Wei DONG ; Xiaoliang JIANG ; Xing LIU ; Zhiwei YANG
Chinese Journal of Comparative Medicine 2015;(5):24-28,19
Objective To determine whether dopamine D5 receptor (D5R) regulates the development of dilated cardiomyopathy (DCM) by inhibiting oxidative stress.Methods We developed heart-specific hD5 mutant (α-MHC-hD5F173L) and wild type (α-MHC-hD5WT) transgenic mice.The NOX2 expression and ROS production were tested in the transgenic mice at three month of age.The α-MHC-hD5F173L mice were treated with either NADPH oxidase inhibitor Apocynin (1mmol/kg/day) or phosphate-buffered saline (PBS) as control by intraperitoneal injection for 4 weeks.After then, the indexes of heart function were measured.The hD5WT and hD5F173L were transfected respectively in rat H9C2 cells, in which ROS production and NOX2 expression were detected at basal level.Results The ROS production and NOX2 expression were higher in the heart of α-MHC-hD5F173L than α-MHC-hD5WT mice.Apocynin treatment improved the heart function of α-MHC-hD5F173L mice.NOX2 expression and ROS production were higher in hD5F173L than hD5WT transfected H9C2 cells.Conclusions Dopomine D5 receptor may prevent DCM development by inhibiting oxidative stress.
10.Study of Plasma amino acid related metabolites of septic rats using gas chromatography-mass spectrometry
Jin SU ; Yimin ZHU ; Yu JIANG ; Lianhong ZOU ; Xiaoliang LIU ; Yixiao XU
Chinese Critical Care Medicine 2017;29(4):332-336
Objective To reveal the pathophysiological changes of sepsis, the plasma metabonomics of septic rats was determined and differential metabolites were determined by gas chromatography-mass spectrometry (GC-MS). Methods Male Spraguee-Dawley (SD) rats about 8 weeks were randomly divided into sham group (n = 18) and sepsis group (n = 24). Cecal ligation and puncture (CLP) was used to build sepsis model, while cecum was kept intact only in the sham group. 6 hours after the operation, rats were anesthetized, and blood was harvested through heart thoracotomy. Then the plasma metabonomics was detected by GC-MS and metabolic profile analysis was performed to find the relative differential metabolites.Results Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) showed that the metabolic profiling of the sepsis group was significantly different from the sham group. 259 kinds of metabolites were got by GC-MS, and 69 kinds of differential metabolites were found between sham group and sepsis group, in which 23 differential metabolites were related to amino acid metabolism. Compared with sham group, the contents of putrescine, N-glutamic acid, hydroxynorvaline, 3-cyanuric acid, D-alanyl-D-alanine and urea in the plasma of septic rats increased significantly, which ratios of sepsis/sham group were 10.876, 6.394, 2.800, 2.226, 1.323, 1.203, respectively (allP < 0.05). On the other hand, the contents of oxygen generation of proline, citrulline, glutamine, su-beta-hydroxy aspartic acid, citric acid, N-methyl-DL-alanine, serine, lysine, threonine, N-formyl-L-methionine, methionine, alanine, nicotinuric acid, N-methyl-L-glutamic acid, trans-4-hydroxy-L-proline, proline, L-glutamic acid in the plasma of septic rats decreased significantly, which ratios of sepsis/sham group were 0.858, 0.853, 0.834, 0.816, 0.816, 0.814, 0.813, 0.801, 0.793, 0.792, 0.774, 0.766, 0.748, 0.727, 0.716, 0.674, 0.603, respectively (allP < 0.05).Conclusions Through the GC-MS analysis of plasma metabonomics of septic rats, we found abnormal energy metabolism changes. The content of amino acid in plasma might be a method to evaluate the energy metabolism status of sepsis.