2.Expression of von Willebrand factor-A1 domain in E coli and it's the inhibitory effect on platelet aggregation
Huaiping ZHU ; Yingchun WANG ; Xia BAI ; Shundong JI ; Wei ZHANG ; Bojing SHAO ; Mingqing ZHU ; Changgen RUAN
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To further investagate the mechanism of thrombus formation and develop a new remedy of anti-thrombus formation. METHODS: The amplified DNA fragment of vWF-A1 domain was inserted into expression vector with 6?his taq (pQE-31), the recombinant expression vect or was transformed into E coli (strain M15) and induced by IPTG. The recombinant fragment, comprising residues 449-728 of mature vWF subunit, designate rvWF-A1. It was purified by Ni-NTA agarose column and renatured by Tris buffer containin g GSH and GSSG. FACS and platelet aggregometer were employed to analyse the rvWF -A1 function of binding to platelet glycoprotein Ib and inhibiting ristocetin-in duced platelet aggregation. RESULTS: The rvWF-A1 was expressed successfully in E coli, comin g up to 30% of total bacterial protein. Its purify was over 95% through Ni-NTA a garose. It was identified to have ability to bind to GPIb, its biologic activity to inhibit ristocetin-induced platelet aggregation was observed, and the inhibi tive rate was 84 7%. CONCLUSION: The above results indicated that high-level expressi on of rvWF-A1 was successfully achieved in E coli and rvWF-A1 may be an effectiv e antithromotic agent in preventing thrombus formation.
3.The alteration of extracellular sodium and potassium ionic activities of hippocampal neurons in epileptiform rats kindled by coriaria lactone
Fei ZOU ; Hong XIE ; Shiming ZHONG ; Na GAO ; Ajing WANG ; Changgen ZHU
Chinese Journal of Pathophysiology 2000;0(08):-
AIM and METHODS: The sodium ion(Na +) and potassium ion(K +) selective microelectrodes were used to measure changes of ionic activity of extracellular sodium and potassium([Na +] o, [K +] o) in hippocampus and hippocampal slice during epieptic seizure induced by intrahippocampal microinjection of coriaria lactone(CL) in rats and perfusing hippocampal slice with CL. RESULTS:30 s, 1min and 2min after injection of CL into hippocampus, the [Na +] o decreased 27.7 mmol/L, 50.3 mmol/L, 57.8 mmol/L respectively and the [K +] o increased 2.3 mmol/L, 2.4 mmol/L, 2.9 mmol/L respectively compared with control values( P
4.CT-guided percutaneous intervention combined with rehabilitation integration treatment for lumbar disc herniation
Dechun DAI ; Guohai TONG ; Lianlong BIAN ; Chunlin TANG ; Weifang ZHU ; Linfeng MEI ; Changgen SHI ; Hao JIANG ; Fangjie HANG
Chinese Journal of Tissue Engineering Research 2013;(48):8449-8454
BACKGROUND:CT-guided percutaneous intervention and rehabilitation techniques are both classic programs for diagnosis and treatment of lumbar disc herniation. Is the combination of CT-guided percutaneous intervention and rehabilitation techniques preferential y used for treatment of lumbar disc herniation?
OBJECTIVE:To evaluate the curative effect of CT-guided interventional injection combined with rehabilitation integration treatment for lumbar disc herniation and to analyze prognostic factors.
METHODS:Eighty-eight patients with lumbar disc herniation were subjected to CT-guided interventional injection combined with rehabilitation integration treatment from May 2010 to May 2013. Injection medicine consisted of betamethasone, tanshinone Ⅱ A sulfonate, neurotropin, normal saline and iohexol. Rehabilitation integration treatment included traction, manipulation, acupuncture, transcutaneous electrical nerve stimulation and thermal magnon. Macnab criteria and Chinese version of Oswestry low back pain disability questionnaire were used to assess the curative effects in comparison with the 112 patients receiving only CT-guided interventional injection that were reported previously. The prognostic factors, such as age, disease course time and herniation type of target segment were testified by correlation analysis.
RESULTS AND CONCLUSION:Total y 77 patients were completely fol owed up for 1 year. There were excellent outcomes in 64 cases, while favorable outcomes in 7 cases, fair outcomes in 5 case, poor outcome in 1 case, with a better outcome rate of 92%.There was a significantly decreased trend in Oswestry disability Index scores at 1, 6, 12 months during the fol ow-ups (P<0.01). No severe complications occurred in al the included patients. The curative effects were improved in term of better outcomes rate compared with the 112 patients receiving only CT-guided interventional injection, but there was no significant difference (P>0.05). The young group (≤ 45 years) had better outcomes than the older group (>45 years;P<0.01). In addition, disease course time and herniation type of target segment were not statistical y significant risk factors predicting clinical results (P>0.05). These findings indicate that CT-guided interventional injection combined with rehabilitation integration treatment could relieve lower back pain and radical leg pain effectively and decrease life disability level.
5.Platelets promote the enhanced tumor metastasis of bone marrow mesenchymal stem cells
Zhuqian LI ; Qianqian WANG ; Changgen XU ; Hongge JI ; Yilong LU ; Xiangdong ZHAO ; Rong QIU ; Li SUN ; Bin CHEN ; Mei WANG ; Wenrong XU ; Wei ZHU
Chinese Journal of Clinical Laboratory Science 2018;36(2):148-151
Objective To investigate the effect of human bone marrow mesenchymal stem cells (BM-MSCs) stimulated by platelets in vitro on the metastasis of cancer cells.Methods The BM-MSCs were isolated and cultured in vitro and platelets from the peripheral blood of healthy persons were purified.The MSCs (control),platelets + MSCs,and platelets treated with culture media (CM) of SGC-7901 tumor cells + MSCs (T-platelets + MSCs) were cultured,respectively,and the MSCs and supernatants (MSCs-CM and SGC-7901-CM) were collected,respectively,after 24 hours.The expressions of markers of cancer-associated fibroblasts (CAF),such as α-SMA and Vimentin,were determined by Western-blotting.The immigration ability of BM-MSCs were analyzed by Transwell assay.The levels of P-selectin in platelets stimulated by MSCs-CM or SGC-7901-CM were detected with flow cytometry.The metastasis model of gastric cancer SGC-7901 cells was established in BALB/c nude mice by the injection of tail vein,and the tumor metastasis in vivo was observed.Results The expression levels of P-selectin in platelets stimulated by MSCs-CM ([21.37 ± 1.00] %) or SGC-7901-CM ([31.4 ± 1.71] % were significantly higher than that in the control ([3.17 ± 0.40] %,t =27.85 and 29.18,P < 0.01).The expression levels of α-SMA and Vimentin in platelets + MSCs group (0.79 ± 0.08 and 0.88 ± 0.01) and T-platelets + MSCs group (0.90 ±0.06 and 0.96 ±0.04) were significantly higher than that in the control (0.64 ±0.02 and 0.75 ±0.05,t =2.96 and 6.45 forα-SMA,t =4.73 and 5.73 for Vimentin,P <0.01).The amounts of immigration cells in platelets + MSCs group (340.3 ±27.7) and T-platelets ± MSCs group (424.3 ± 17.6) were significantly higher than that in the control (220.7 ± 19.4,t =6.14 and 13.48,P < 0.01).The in vivo experimental results showed that the metastatic foci in platelets ± MSCs group (4 ± 2) and T-platelets ± MSCs group (21 ± 4) were significantly higher than that in the control (0.33 ± 0.06,t =3.051 and 8.857,P < 0.01).Conclusion Platelets promote the immigration and the enhanced tumor metastasis in vivo of BM-MSCs.