1.The effects and mechanism of SOCS3 gene transfection in CD4+Th cell differentiation and expression of inflammatory cytokines of mouse
Pei ZHANG ; Nianguo DONG ; Jinping LIU
Chongqing Medicine 2016;45(36):5049-5051,5055
Objective To investigate the effect and mechanism of adenovirus vector mediated SOCS 3 gene transfection in CD4+ Th cell differentiation and expression of inflammatory cytokines of mouse .Methods The CD4+ Th cells were isolated from spleen of C57bl/6 mouse and cultured .Ad‐SOCS3 were transfected into the CD4+ Th cells .PHA was used for culturing with the CD4+ Th cells .RT‐PCR were used to detect the mRNA expression ,and Western blot were used to detect the protein expression of cytokines .Results Compared with the control group ,the gene and protein expression of T‐bet ,IL‐2 ,IFN‐γ,STAT4 and IL‐12Rβ2 in the transfected group were significantly down‐regulated ,the gene and protein expression of SOCS3 ,GATA‐3 ,IL‐4 ,IL‐6 ,IL‐10 and STAT6 were significantly up‐regulated(P<0 .01) .Conclusion The results indicate that SOCS3 gene transfection can up‐regu‐late SOCS3 mRNA and protein expression in the CD4+ Th cells ,down‐regulate the JAK/STAT pathway ,inhibition of Th1 cell dif‐ferentiation ,and down regulation of inflammatory cytokine gene and protein expression ,and indirectly promote Th2 cell differentia‐tion ,and up the corresponding inflammatory cytokine gene and protein expression .
2.Effects of Xinshang Xuduan Decoction on Osteoblast Proliferation and Bone Morphogenetic Protein in Vitro
Pei ZHANG ; Lujue DONG ; Yikai LI
Journal of Guangzhou University of Traditional Chinese Medicine 2016;(1):92-97
Objective To investigate the effects of Xinshang Xuduan Decoction(XXD, mainly composed of Rhizoma Drynariae, Radix Dipsaci, and Eupolyphaga seu Steleophaga) on the proliferation and differentiation of rat osteoblasts and bone morphogenetic protein 2(BMP-2)expression in the osteoblasts . Methods The animal serum containing XXD was prepared by serum pharmacological method and then was mixed together with α-MEM for the cell culture. Osteoblasts were isolated from the skull bone of SD neonatal rats by collagenase digestion and were identified by alkaline phosphatase(ALP) staining and alizarin red staining. The third and fourth generations of osteoblasts were treated with XXD at the volume fraction of 5%, 10%, 20% for 24, 48 , 72 hours respectively, and then the proliferation of the cells was evaluated by Cell Counting Kit-8(CCK-8). In the other test, osteoblasts were cultured with blank control serum, and 10% serum containing XXD, Radix Dipsaci, and Rhizoma Drynariae, respectively, and then the ALP activity was examined by using ALP assay kit and the expression of BMP-2 was investigated by enzyme-linked immunosorbent assay(ELISA). Results XXD had a dose- and time-dependent effect on the proliferation of rat osteoblasts in a suitable volume fraction range from 5% to 20%, and the effect of XXD at 10% was the best. Compared with the blank control serum group, ALP activity was increased in the cells treated with 10% serum containing XXD, Radix Dipsaci, and Rhizoma Drynariae(P<0.01). On culturing day 7, the expression of BMP-2 was increased in 10% XXD group and Rhizoma Drynariae group(P<0.05). Conclusion XXD can increase the ALP activity and BMP-2 expression in the osteoblasts in vitro, so does the single herb of Rhizoma Drynariae. And their therapeutic mechanism in promoting the healing of fractures may be related with the enhancement of osteogenesis of osteoblasts.
3.Bone morphogenetic proteins and tumors
Dong MI ; Yuexiang ZHANG ; Shuqin CHEN ; Pei QI
Journal of International Oncology 2013;(5):323-325
Bone morphogenetic proteins (BMPs) are multi-functional growth factors,they are found not only inducer of new bone formation,but also have critical roles in cell growth,differentiation,migration,apoptosis,embryogenesis and organogenesis.Studies show that BMPs are closely related to tumorigenesis and metastasis,which have important practical values in tumour therapy.
4.Autophagy inhibits PDGF-BB-induced calcification in vascular smooth muscle cells
Qianqian PEI ; Han MEI ; Xuhui ZHANG ; Lihua DONG
Chinese Journal of Pathophysiology 2016;32(8):1506-1506,1507
AIM:To investigate the relationship between autophagy and calcification in vascular smooth muscle cells ( VSMCs) after platelet-derived growth factor (PDGF)-BB stimulation.METHODS:Cultured VSMCs were stimulated with PDGF-BB for different time, the expression of vascular calcification-related proteins and autophagy-related proteins were detected by Western blot .The interaction be-tween Beclin1 and PI3KC3 was detected by co-immunoprecipitation.RESULTS: The expression of BMP2 and ALP showed a trend from decline to rise.ALP slumped at 12 h, and BMP2 slumped at 6 h.Moreover, the expression of Beclin-1 showed a trend from rise
to decline, and peaked at 12 h.The conversion of LC3-ⅠtoⅡincreased in a time-dependent manner , and peaked at 24 h.The ex-pression of BMP2 and ALP was increased in VSMCs incubated with PDGF-BB and autophagy inhibitor 3-MA, compared with PDGF-BB-stimulated VSMCs.Furthermore, the interaction between Beclin1 and PI3KC3 was enhanced at 6 h after PDGF-BB stimulated, peaked at 12 h, and kept in high level at 24 h.Moreover, the phosphorylation level of Beclin 1 was enhanced by PDGF-BB stimulation, and peaked at 6 h.CONCLUSION:Our findings demonstrate that PDGF-BB-induced autophagy inhibits VSMC calcification by en-hancing Beclin1 phosphorylation and interaction between Beclin 1 and PI3KC3.
5.Use of posterior or anterior renal lip incision for partial nephrectomy in the treatment of renal hilum endophytic renal cell carcinoma
Zhiling ZHANG ; Yonghong LI ; Pei DONG ; Zhuowei LIU ; Fangjian ZHOU
Chinese Journal of Urology 2015;36(3):172-174
Objective To introduce our experience of performing posterior or anterior renal lip incision for partial nephrectomy in the treatment of renal hilum endophytic renal cell carcinoma.Methods From Jan.2010 to Jan.2014,five female patients with renal hilum tumors were treated in our institute.The median age was 54 (51-72) years.The median tumor diameter was 4.0 (2.8-4.8) cm.The median preoperative creatinine was 53.9 (52.6-75.4) μmol/L.One of them was solitary kidney with absolute indication; three cases had basic disease with relative indication; one was with selective indication.The patients were put in supine or lateral position.After general anesthesia,we preformed partial nephrectomy by cutting posterior renal lip in 3 cases and the anterior lip in 2 cases.We clamped the renal artery,opened the renal posterior or anterior lip,then dissected the tumor beside the pseudo-capsule.After removing the tumor,we used 3-0 absorption suture to control bleeding and repair the opened collecting system.Finally,we used 2-0 absorption suture to close the renal defect.Results The median operation time was 195 (155-215) min; the median renal warm ischemia time was 35 (15-70) min; the median postoperative hospital stay was 8 (7-9) d.There was no secondary bleeding and urine leakage happened.The pathological results showed that 3 cases with clear cell carcinoma,1 with papillary carcinoma and 1 with renal medullary interstitial cell tumor.All patients showed normal kidney shape.The median postoperative creatinine was 63.0 (59.4-75.4) μmol/L.After a median follow up of 24.2 mon,all patients survive without tumor recurrence.Conclusions The short-term result of posterior or anterior renal lip incision partial nephrectomy in treating endophytic renal hilum endophytic renal cell carcinoma is safe and feasible.
6.Expression of Death Receptor 4 and Death Receptor 5 in Human Craniopharyngioma
pei-dong, LI ; zhong-wei, ZHAO ; xin-jun, WANG ; dong-ling, GAO ; yun-han, ZHANG
Journal of Applied Clinical Pediatrics 2004;0(11):-
Objective To investigate the expression and significance of death receptor 4(DR4) and DR5 in human craniopharyngioma.Methods The expression of DR4 and DR5 was determined by immunohistochemistry and in situ hybridization in 28 samples of craniopharyngioma and 25 samples of normal brain tissue.Results With low expression in partial normal brain tissue,DR was expressed highly in all of the craniopharyngioma samples.High DR expression in craniopharyngioma tissue differed from low DR expression in normal brain tissue(P0.05).Conclusions High DR expression is prevalent in craniopharyngioma tissue.This may contribute to the apoptosis-induced therapy of craniopharyngioma.The control of DR expression lays in protein level.This may contribute to the selective induced-apoptosis of tumor necrosis factor-related apoptosis-induced ligand.
8.Heat-killed Staphylococcus aureus induces a common early response in human monocytes in the presence of high concentration glucose
Ying CHEN ; Yan ZHANG ; Xin SONG ; Pei SUN ; Bai CHANG ; Haidong LI ; Dong MENG ; Qiaofen LI
Chinese Journal of Microbiology and Immunology 2012;(12):1005-1010
Objective To investigate the effects of heat-killed Staphylococcus aureus (HKSA) on the apoptosis and expression of iNOS and IL-1β in THP-1 monocytes in the presence of high concentration of glucose.Methods THP-1 cells were cultured in medium containing 25.0 mmol/L(HG) or 5.5 mmol/L (LG,control) D-glucose for 12 h-8 d.The THP-1 cells cultured for 6 d were extracted on the 0-48 h with or without HKSA,then apoptosis and expression of iNOS and IL-1β were examined.Apoptosis was analyzed by flow cytometry and expressions of IL-1β and iNOS were quantitated by real-time PCR.Results The expression of iNOS and IL-1β in THP-1 monocytes was increased significantly in the presence of high concentration of glucose for 12-48 h(P<0.05),reaching the highest level at 24 h and returned to baseline after 4 d.The expression was significantly lower than that of control after 4-6 d.Apoptosis rate was also increased significantly after 48 h to 4 days.HKSA infection enhanced apoptosis,but inhibited the expression of iNOS and IL-1 β in the presence of high concentration of glucose.The expression of iNOS and IL-1β increased significantly at 6 h(P<0.01),reaching the highest level at 12 h,but the levels were significantly lower than those in control groups (P<0.05).Conclusion These data suggest that high concentration of glucose can interfere with the anti-bacterial function of monocytes by reducing their expression of iNOS and IL-1β and enhancing their apoptosis.
9.MRI diagnosis of Hirayama disease
Yu FU ; Dong-Sheng FAN ; Xin-Long PEI ; Hong-Bin HAN ; Jun ZHANG ; De-Xuan KANG ;
Chinese Journal of Radiology 2001;0(04):-
Objective To investigate the dynamic changes of the spinal cord during neck flexion in Hirayama disease for diagnosis.Methods MRI examinations in neutral neck position and a fully flexed neck position were performed on 18 cases of Hirayama disease and 31 young normal control subjects.We measured an antero-posterior diameter(APD)and transverse diameter(TD)of the cervical cord at the superior margin of the C6 vertebral body for each position,and investigate the dynamic changes.The different in frequency of these findings between the control and patient groups was examined by means of the x~2 test.The group means were compared by independent-sample t-test.Significance was defined as P
10.Effect of sodium arsenite exposure on DNA damage of rat peripheral blood lymphocytes
Feng-jie, TIAN ; Yuan, XU ; Xiang-dong, GAO ; Yan-ning, ZHANG ; Yi, GAO ; Qiu-ling, PEI
Chinese Journal of Endemiology 2011;30(1):13-15
Objective To explore the DNA damage in peripheral blood lymphocytes of rats exposed to sodium arsenite. Methods Thirty-two Wistar rats, weighing 180 - 200 g, equal male and female, were randomly divided into 4 groups, 8 in each group. Sodium arsenite 0(control) ,0.05,0.15,0.45 mg/L were given through drinking water for 30 days. Body weight and drinking water consumption were measured every day. Blood were collected and DNA damage in peripheral blood lymphocytes was examined by single cell gel electrophoresis.Results The increase of body mass[( 121.00 ± 38.57), ( 120.62 ± 42.80), ( 125.38 ± 48.68)g]and water intake [(36.9 ± 6.2), (37.9 ± 5.8), (39.3 ± 4.2)ml/d]in 0.05,0.15,0.45 mg/L sodium arsenite groups were compared with the control group[( 119.25 ± 47.27)g, (38.4 ± 5.1 )ml/d], and the difference were not significant (F = 0.040,0.828, all P > 0.05). The tail ratios[46.25%(185/400) ,57.00%(228/400),64.00%(256/400)], tail lengths [(32.89 ± 17.18), (58.74 ± 36.28), (77.55 ± 35.73 ) μm]and tail moments [(6.29 ± 3.74), ( 11.20 ± 9.64),(17.30 ± 12.60)μm]in 0.05,0.15,0.45 mg/L sodium arsenite groups were significantly higher than those of the control group[39.25%(157/400), (18.73 ± 15.83),(2.61 ± 1.05)μm, all P < 0.01], and the tail ratios,tail lengths and tail moments in lymphocytes increased with increased doses of arsenic concentration. Conclusions Low doses of arsenic exposure can induce DNA damage in peripheral blood lymphocytes of rats.