1.The Apoptosis Induction Effect of Recombinant Caspase-3 on the Human Osteosarcoma Cell Line SOSP-9901
Bing YU ; Qingyu FAN ; Lu YAN
Journal of Chinese Physician 2000;0(12):-
Objective To investigate the apoptosis induction effect of recombinant caspase-3 expression on osteosarcoma cell line SOSP-9901. Methods Recombinant caspase-3 gene was subcloned into the GFP reporter vector pEGFP-C1 to generate the expression vector pEGFP-caspase-3 by DNA recombinant technique. pEGFP-caspase-3 was transfected into human osteosarcoma cell line SOSP-9901 by Lipofectamine 2000. The expression of recombinant caspase-3 was determined by RT-PCR. The morphological changes of transfected cells were observed under flurescent and electronic microscope. The cell survival rate of transfected cells was assayed by MTT method. Results Recombinant caspase-3 gene could be stably expressed in the transfected SSOP-9901 cells. Recombinant caspase-3 could obviously induce SOSP-9901 apoptosis, and inhibit SSOP-9901 cell proliferation in vitro. Conclusion Recombinant caspase-3 could inhibit the growth of the osteosarcoma cell line SOSP-9901 and induce it into apoptosis.
2.The role of Notch3 in tumorgenesis and progression
Jie QIAO ; Caijie LU ; Qingyu ZHANG
Practical Oncology Journal 2014;(5):451-455
Notch signaling pathway is a highly conserved signaling pathway ,and plays an important role in the individual growth and development process in the vertebrate .Many studies have demonstrated Notch 3 re-ceptor expression is aberrant in a variety of tumors .Recent studies show that aberrant Notch 3 mediated signaling pathway is closely related with the progression of diverse types of tumors .Notch3 plays a vital role in apoptosis , proliferation and differentiation of tumor cells .To promote the studies of Notch 3 as a drug development targets ,the molecular mechanisms of abnormal Notch 3 expression and its role on tumorgenesis ,progression and treatment re-sistance are reviewed in this paper .
3.Effect of Protobothrops mucrosquamatus venom and its fractions on bIood coaguIation
Yanan LL ; Qianyun SUN ; Qingyu LU
Chinese Journal of Pharmacology and Toxicology 2015;(2):284-290
OBJECTIVE To investigate the effect of Protobothrops mucrosquamatus venom (PMV) and its fractions on functions of the circulatory system in vitro in order to better understand its toxicity mechanism. METHODS PMV was isolated to three fractions FⅠ, FⅡ and FⅢ with a different molecular mass range by Sephadex G-75 gel filtration chromatography. Platelet rich plasma was adjusted to 3×1011 L-1 by platelet poor plasma. Platelet suspension was incubated with PMV and its fractions 0.03 g.L-1 for 5 min, respectively, and platelet aggregation was determined on an LBY-NJ4 aggregometer. PMV and its fractions 0.05 g.L-1 were preincubated with plasminogen 0.1 U.L-1 for 10 min before chromogenic substrate cleavage activity was measured by endpoint and enzyme kinetics determination. PMV and its fractions 1.0 g.L-1 were incubated with rat plasma for 5 or 30 min, and thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT) and fibrinogen (FlB) content were assayed. The microvascular endothelial cells were exposed to PMV and its fractions 10, 50 and 250 mg.L-1 , respectively, for 24 h, while the morphological change was observed using an inverted phase contrast microscope, and the cell viability was determined by MTT method. PMV and its fractions were incubated with guinea pig red blood cell suspension in the presence or absence of lecithin for different time, and hemolysis was measured. RESULTS Compared with normal control, platelet aggregation rate was significantly increased by PMV and FⅠ (>71 ku)〔(12.4±4.1)%,(61.0±5.8)% and (56.9±5.9)%〕(P<0.01). PMV and FⅡ (18-37 ku) significantly hydrolyzed chromogenic substrate S-2251(P<0.01). PMV and FⅠ caused plasma coagulation. Compared with normal control, FⅡ and FⅢ (<10 ku) remarkably prolonged TT, APTT and PT( P<0.01). Morphological observation revealed that PMV, FⅠ and FⅡdetached the adherent cells. Compared with normal control group, PMV, F Ⅰ and F Ⅱ inhibited cell viability, and the survival rate of the cells decreased to (56.8±3.6)%,(71.6±3.8)% and(58.2±5.5)%, respectively. PMV and FⅡ slowly caused slight hemolysis in absence of lecithin. PMV and FⅡ caused significant hemolysis in the presence of lecithin, and the hemolytic rate increased to (81.0±4.0)% and (81.0±1.0)%( P <0.01) in 0.5 min, respectively, compared with (17.7±1.0)% of the control group. CONCLUSION PMV possesses different activities that affect the functions of the circulatory system in vitro, and the fractions play different roles in toxicity mechanisms.
4.Effect of Hedyotis Diffusa Extract on Apoptosis of Human Hepatocarcinoma Cell Line Bel7402
Xiufang LU ; Qingyu MENG ; Xinmin GUO
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(11):-
Objective To study the inducing apoptosis of Hedyotis diffusa extract (HDE) on human hepatocarcinoma cell line Bel7402 and its molecular mechanism. Methods Human hepatocarcinoma cell line Bel7402 was used in vitro cell culture, there were 3 groups:control group, HDE group and 5-Fu group, the Bel7402 cells were dealed with them respectively. The apoptosis morphologic changes of human hepatocarcinoma cells were observed by invert microscope. The apoptosis rate was detected with hematoxylin stain by light microscope. Inhibition of Bel7402 cell proliferation was measured by MTT assay. The expression of oncogene Bcl-xl and anti-oncogene p53 of Bel7402 ceils were observed with RT-PCR assay. Results Compared with the control group, the condensation of nuclear, vacuolar degeneration of mitochondria could be found through light microscope. The apoptosis rate of HDE group was remarkably increased compared with that in control group (P
5.The use of cutting ballon in small vessel diffuse lesion interventions
Jianghong LI ; Qingyu LU ; Ling ZHANG
Chinese Journal of Interventional Cardiology 2003;0(06):-
Objective To evaluate the efficacy and safety of cutting ballon angioplasty (CBA) in small vessel diffuse lesion Methods 44 patients with 48 lesions underwent successful coronary angioplasty with cutting ballon angioplasty Results All of the lesions were treated successfully by 4 6?2 4 times dilation of cutting ballon The total duration of ballon inflation was (227 4?102 3) s The lesions were dilated at a pressure up to (901 8?222 9) kPa The severity of vascular stenosis lessened obviously [(88 6?7 9)% vs (16 8?15 4)%, P =0 001] without severe complications Conclusion Cutting ballon angioplasty (CBA) on small vessel diffuse lesion is safty and effective
6.Clinical application of thrombus aspirated catheter (DIVER~(TM)) in primary PCI for acute myocardial infarction
Jianguo WU ; Ling ZHANG ; Qingyu LU
Chinese Journal of Interventional Cardiology 1993;0(02):-
Objective To evaluate the clinical effect and safety of the thrombus aspirated catheter (DIVER~(TM)) in primary PCI for acute myocardial infarction. Methods Seventy four AMI patients with total occluded infarct-related coronary artery(IRA)and intra-coronary thrombus from March 2005 to April 2006 were divided into two groups according to admission time and the treatment they received. After the 0.014 in BMW coronary wire crossed the lesion, the thrombus aspirated catheter (DIVERTM) was advanced over the wire to the lesion and continuously aspirated 2-3 times until the thrombus disappeared and stents were directly implanted in patients in group A (n=36) admitted from December 2005 to April 2006. Routine PCI applied in patients in group B (n=38) admitted from March 2005 to December 2005. Sirolimus eluting stents were implanted in both groups. The artery re-opened rate, no-reflow rate and the rate of major adverse cardiac events during hospitalization and 6 months after discharge were compared between two groups. Results Coronary arteriography showed the artery re-opened rate was 100% in both groups. The rates of no-reflow and major adverse cardiac events during hospital stay were significantly lower in group A than those in group B (2.78% vs 21.05% and 2.78% vs 10.53% respectively,P
7.Expression of coagulation-and fibrinolysis-related molecules of endothelial cells induced by activated complement alternative pathway and intervention
Qingyu LU ; Min LI ; Qianyun SUN
Chinese Pharmacological Bulletin 2015;(8):1142-1146
Aim To investigate the change of molecu-lar expression related to coagulation and fibrinolysis in human microvascular endothelial cells ( HMEC ) in-duced by activated complement alternative pathway and effect of pyrrolidine dithiocarbamate ( PDTC ) and res-veratrol on intervention. Methods Normal human se-rum was activated by cobra venom factor ( CVF) . After exposure of HMEC to activated complement for various times, supernatant was removed and assayed for ex-pressions of P-selectin, VWF, t-PA, PAI-1, TF, TM, and NO by using reagent kits. The expressions of the above molecules in HMEC pretreated with PDTC and resveratrol were also investigated. Results P-selectin and VWF were rapidly released by endothelial cells and the expression reached the peak at the time point of 15 min. The expressions of t-PA, PAI-1, and TF were continuously upregulated, whereas NO and TM were decreased. PDTC and resveratrol inhibited the upregulation of P-selectin, VWF, t-PA, PAI-1 and TF, and intervened the downregulation of NO. Res-veratrol further downregulated the expression of TM. Conclusion Activated complement alternative path-way can influence the expression of molecules related to coagulation and fibrinolysis in HMEC, and PDTC and resveratrol can affect this change.
8.Effects of hyaluronic acid on scar formation in the acellular nerve allograft
Yingwei LIU ; Wanli ZHANG ; Chengtao CHI ; Qingyu XU ; Dezhi LU
Chinese Journal of Tissue Engineering Research 2016;20(42):6317-6323
BACKGROUND:In terms of the histocompatibility, immune rejection and scar formation after repair, acel ular nerve al ograft is closer to autologous nerve cel s. At present, hyaluronic acid has been applied for autologous peripheral nerve repair;however, research on the nerve al ograft is rarely reported.
OBJECTIVE:To explore the effect of hyaluronic acid on the anastomotic scar in acel ular nerve al ograft repair of rat sciatic nerve defect.
METHODS:Thirty-six Sprague-Dawley rats were randomly divided into three groups (n=12 per group). The rat model of nerve defect of 10 mm was established by cutting the sciatic nerve of the left hind leg and then given nerve al ograft combined with the injection of hyaluronic acid at anastomosis (experimental group), only nerve al ograft (control group) and autologous nerve graft (nerve autograft group), respectively. Afterwards, the healing of the proximal anastomosis was observed and scar components were assessed.
RESULTS AND CONCLUSION:Gross observations showed that the rat skin and muscle fascia had no significant differences in healing among groups, while the surrounding tissue adhesion in the experimental group was milder than that in the control group (P<0.05). Masson staining found that col agen deposition in the epinerium could be observed in each group. In the experimental group, a smal amount of col agen fibers arranged orderly in the epineurium;in the control group numerous col agen fibers accumulated and arranged irregularly;in the nerve autograft group, sparse epineurial col agen fibers appeared in an order arrangement. The gray value of col agen type I in the experimental group was higher than that in the control group (P<0.05), while the gray value of col agen type III was lower than that in the control group (P<0.05). No significant differences were found in the sum gray values of col agen type I and III among groups (P>0.05). These findings indicate that in the peripheral nerve repair, hyaluronic acid abrogates the scar formation by increasing the deposition of col agen type III and reducing the deposition of col agen type I.