2.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.
3.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.
4.C-myc antisense oligodeoxynucleotides transferred by soluble stent preventing vein graft stenosis
Hongtao SUN ; Ying DU ; Qingyu WU
Chinese Journal of Thoracic and Cardiovascular Surgery 1995;0(05):-
Objective A rabbit model of common carotid artery grafted by external jugular veins was used. Vein grafts were transferred by c-myc antisense oligodeoxynucleotides(ODN) and carried by soluble stent. Try to find a new approach to prevent veins graft failure on molecular basis. Methods New Zealand rabbit were randomly divided into five groups, 10 animals each. Pretreating the soluble stents with synthesized c-myc ODN, following stents: (1) control group; (2) soluble stent; (3) soluble stent with sense-ODN; (4) soluble stent with antisense-ODN; (5) soluble stent with mismatch-ODN were put into the vein graft during end to end anastomosis. After 7, 28 and 90 days of operation, vein grafts were harvested. HE and ET stain were made aim to calculate the extent of intima hyperplasia. The expression of c-myc and PCNA were identified by immunchemistry methods. Situ hybridization and Northern Bloting were made to assess the expression of c-myc mRNA quantitatively and semi-quantitatively in veins. Results In the vein grafts of 7,28,90 days post-operation of soluble stent antisense-ODN group ①Intima hyperplasia were inhibited significantly compare with other four groups. ②The expression of c-myc and PCNA were inhibited significantly compare with other four groups. ③C-myc mRNA expression level was significantly Lower than the other four groups. Conclusion Soluble stent can transfer c-myc ODN effectively. C-myc antisense-ODN transferred by soluble stent can inhibit the expression of c-myc and PCNA, can inhibit intima hyperplasia of vein graft significantly, thus preventing the stenosis of vein graft.
5.Nosocomial Infection and Risk Factors in Our Cancer Center 2006-2007
Qingyu ZHAO ; Yueli SUN ; Yuchan LIANG
Chinese Journal of Nosocomiology 2009;0(17):-
OBJECTIVE To study the incidence rate of nosocomial infection and risk factors in our Cancer Center during 2006-2007.METHODS All the reported cases of nosocomial infection from 2006 to 2007 in our Center were analyzed retrospectively.RESULTS From the 50011 hospitalized patients,952 were infected.The infection rate of the hospital was 1.9%,with the case infection rate of 2.12%.Infection usually occurred at the respiratory tract,gastrointestinal tract and oral cavity,with the constituent ratio of 48.6%,34.12% and 13.22%,respectively.Cancer,chemotherapy,radiotherapy and surgery were the leading predisposing factors.CONCLUSIONS The main infected site is respiratory tract.Cancer and the clinical therapies may lead to infection,so we should pay more attention to the nosocomial infection in the cancer therapy.
6.Abnormality of coagulation and fibrinolysis functions of endothelial cells induced by activated complement alternative pathway and intervention
Min LI ; Qingyu LU ; Yanan LI ; Qianyun SUN
Chinese Pharmacological Bulletin 2017;33(10):1430-1435
Aim To investigate the change of coagulation and fibrinolysis functions of human microvascular endothelial cells (HMEC) induced by activated complement alternative pathway and effect of pyrrolidine-dithiocarbamate (PDTC) and resveratrol (Res) on intervention.Methods Normal human serum was activated by cobra venom factor (CVF).After exposure of HMEC to activated complement for various time points, supernatant was removed and assayed for activities of hydrolysing chromogenic substrate and affecting activated partial thromboplastin time (APTT) and prothrombin time (PT).The cells exposed to activated complement were collected and washed, and then the cell suspension was assayed for activity of affecting coagulation function of normal plasma.Lastly, the coagulation and fibrinolysis functions of HMEC pretreated with PDTC and Res were also investigated after HMEC was exposed to activated complement alternative pathway.Results The hydrolysis activity of chromogenic substrate of supernatant was up-regulated significantly after HMEC exposed to activated complement alternative pathway.The supernatant induced APTT decreased significantly, and also shortened PT.The cell suspension of various time points induced APTT decreased significantly, and also shortened PT by suspension of 6 h time point.PDTC and Res failed to inhibit the up-regulation of the chromogenic hydrolysis activity, but Res showed significant intervention on decrease of APTT, and PDTC had better effect on inhibiting the decrease of PT than that of Res.Conclusion Activated complement alternative pathway can induce abnormality of coagulation and fibrinolysis functions of HMEC, and PDTC and Res can affect this change.
7.Investigation of the frequency of medical radiation exposure in Ningxia region
Yanling SUN ; Guicai LU ; Hongcheng LI ; Qingyu KONG ; Xuli JI
Chinese Journal of Radiological Medicine and Protection 2016;36(10):757-760,779
Objective To understand the allocation of radiological diagnostic equipment and the frequency of medical radiation exposure in the medical institutions in Ningxia region,in order to provide the basis for the reasonable application of medical radiation technology.Methods The general survey in the form of the questionnaires was used to investigate and analyze the medical radiation usage of medical X-ray diagnostics,interventional radiology,nuclear medicine and radiotherapy in Ningxia region medical institutions.Results The frequency of X-ray diagnostic radiography in Ningxia region was 727.9 per 1 000 population in 2014.The frequency of X-ray photography examination was the highest,525.2 per 1 000 population,followed bv the frequency of CT scanning,147.9 per 1 000.For others,the frequency was radiotherapy 6.0 per 1 000 population,nuclear medicine 1.8 per 1 000 and interventional radiology 3.8 per 1 000,respectively.These differences were of statistical significance in frequency of X-ray diagnostic examination in five cities' medical institutions in Ningxia (X2 =162 280.7,P < 0.05),also in all medical institutions at different levels (x2 =902 485.2,P < 0.05) and for both frequency of X-ray photography examination and CT scanning in these institutions (x2 =471 574.9,181 887.1,P < 0.05).Conclusions Of the X-ray diagnostic radiography,the CT scanning has becoine a major means next only to the X-ray photography examination.There are differences in frequency of X-ray diagnostic examination in the medical institutions at different levels in different cities of the region.The related regulatory authorities should strengthen the supervision and management of radiation protection in mnedical institutions,to ensure the justification of diagnostic radiology and radiotherapy.
8.Clinical features and management of combined thoracoabdominal injury
Songchang SUN ; Qingyu ZHANG ; Hongwei ZHANG ; Yong GONG ; Jie LUO
Chinese Journal of General Surgery 1993;0(02):-
Objective To review the clinical features and early management of combined thorcoabdominal injury(CTI),to decrease the mortality rate.Methods A retrospective study was carried out in 63 cases of CTI treated surgically from May 1990 to May 2007.There were 34 cases with blunt injuries and 29 cases with penetrating injuries.Forty-seven patients had rapture of left diaphragm,9 cases had rupthure of right diaphragm,7 cases had rupture of both diaphragms,24 cases were combined with traumatic diaphragmatic hernia,63 cases were combined with hemopneumothorax,and 60 cases were combined with shock.Forty-one cases were diagnosed correctly before surgery.Sixty-three cases were treated by surgery,among them,surgery was performed through left thoracotomy in 18 cases,through a right thoracotomy in 2 cases,through bilateral thoracotomy in 5 cases,through thoraco-abdominal incision in 2 cases,and through the abdominal route in 32 cases.Results Fifty-six cases were cured,and 7 cases(11.1 %) died.The main cause of death was hemorrhagic shock.Conclusions CTI has high mortality rate,and the most frequent cause of mortality is hemorrhagic shock.The operative approach is based on the individual injury condition.Early diagnosis and timely surgical intervention is the key to improving the cure rate and reducing mortality in severe CTI.
10.Fabrication of osteochondral scaffolds containing calcified cartilage zone
Qingyu LIU ; Fuyou WANG ; Junli LIU ; Xinwei SUN ; Liu YANG
Chinese Journal of Trauma 2014;30(5):467-470
Objective To establish osteochondral scaffolds with remained calcified cartilage zone for finding an ideal scaffold for tissue engineered repair of osteochondral defect.Methods Cartilage zone was harvested from fresh adult porcine knee to fabricate type Ⅱ collagen hydrogel.Bone blocks measuring 8 mm in diameter with calcified cartilage zone were prepared by trephine and acellular treatment was performed.Histological staining was used to identify complete removal of cells.To fabricate the osteochondral models containing calcified cartilage zone,type Ⅱ collagen was seeded onto the acellular bone blocks with calcified cartilage zone,lyophilized,and cross-linked with 10 g/L of genipin ethanol solution.Then cell seeding and scanning electron microscope were performed after the models were established.Results HE staining,toluidine blue staining,solid green staining,safranin O staining,and DAPI staining showed cells were completely removed from bone blocks by decellularized process.Porosity of type Ⅱ collagen sponge was (91.1 ±3.8) % and pore size was (79.7 ± 17.1) μm.Porosity of acellular bone blocks was (73.5 ±2.6)% and pore size was (470.2 ± 158.8) μm.Cells seeded onto osteochondral scaffolds grew well by scanning electron microscope.Conclusion Osteochondral scaffolds with calcified cartilage zone provide good biocompatibility and suitable pore size and porosity and may be an ideal material for repairing osteochondral defect in tissue engineering.