ABSTRACT
Objective To investigate the influence of proinflammatory factor interleukin-18(IL-18) on vein endothelial cell function by activating NF-κB mediated cell signal pathway and its association with deep vein thrombosis(DVT).Methods Recombinant human IL-18 was used to act on in vitro cultured human umbilical vein endothelial cell(HUVECs).The NF-κB activation inhibitor was used to conduct interference.The detection measures of real time fluorescence quantitative PCR,Western blot,immunofluorescence and flow cytometry were used to verify whether IL-18 affect the expression of endothelial cellular function markers such as HUVECs normal statusand vWF,P-selectin and tissue plasminogen activator(t-PA) by activating NF-κB mediated cell signal pathway.Moreover the mechanism of IL-18 participating in the DVT was performed the comprehensive analysis by combining with previous study.Results IL-18 could activate NF-κB in endothelial cell,increased the p65 expression in nucleus,decreased the intracellular IκBα expression and significantly increased early apoptosis cells in HUVECs;adding QNZ(EVP4593) could significantly inhibit the activation effect of IL-18 on NF-κB,the occurrence of cellular injury and apoptosis was significantly reduced;IL-18 could promote the abnormal expression of DVT related endothelial cell markers vWF,P-selectin and t-PA (P<0.05).But various markers could recover conventional expression after inhibiting NF-κB activation.Conclusion The interaction between Il-18 and NF-κB causes the abnormality of HUVECs growth status and function,which may be the DVT onset related pathogenic mechanism.
ABSTRACT
Galectins are a family of carbohydrate-binding proteins that have a high affinity to galactosides on cell surfaces and extra cellular glycoproteins.They are involved in a variety of biological functions,including modulation of cell apoptosis,cell activation and inflammation.Our laboratory has recently identified galectin-3 binding protein (Gal-3BP) as being up-regulated in a microparticle proteomics analysis for deep venous thrombosis (DVT) patients compared to negative controls.P-selectin,another glycoprotein involved in thrombus propagation,has been proved as a promising target for DVT management and has been widely studied by our group.Galectins are involved in P-selectin expression and can potentially be implicated in the venous thrombogenesis process.Interleukin-6,as an important cytokine of body,which have been found in the study of inflammation for venous thromboembolism,may be an important factor for the frmation of VT.Research shows gal3bp:gal3 plays a critical role in VT,likely via IL-6 Promote thrombosis.The function of galectins,their role in inflammation and thrombosis as well as their potential implications as a new pharmacological target for DVT management are reviewed in this manuscript.
ABSTRACT
Objective To investigate the correlation between IL‐18 and deep venous thrombosis disease and its clinical significa‐tion .Methods To detect the expression of IL‐18 by ELISA ,we collected the blood samples of DVT patients as the experimental group(n=40) compared to the control group(n=40) and normal group(n=20) .IL‐18 over expression/interference vectors were constructed and transfected human vein endothelial cells ,analyzed by microarray and KEGG Pathway as biology information tech‐nology .Then discuss the association between IL‐18 and DVT .Results Results of ELISA showed that compared with control group and normal group ,the expression of IL‐18 gene in DVT patient were up‐regulated(F=11 .248 ,P0 .05) .Immunofluorescence detected IL‐18 gene expression in cytoplasm of human umbilical vein endothelial cells (HUVECs) .According to the microarray analysis we found in the IL18‐pCDH‐GFP transfected cells 17 signaling pathways were down‐expressed while 16 signaling pathways were up‐expressed .Compared with normal group cells ,in the IL18‐LMP‐shRNAmir1 transfected cells 23 signaling pathways were down‐ex‐pressed and 9 signaling pathways were up‐expressed .Conclusion Based on the above experimental data ,it is very clear that IL‐18 influenced HUVECs and plays an important role in DVT ,it is possible to predict the diagnosis of DVT and act as candidate molecu‐lar markers .
ABSTRACT
Objective To investigate the association between the signaling pathways of MCP-1-pCDH-GFP-trans?fected cells and deep venous thrombosis (DVT). Methods The cultured human umbilical vein endothelial cells (HUVECs) were tested by immunofluorescence and co-immunoprecipitation methods. The constructed MCP-1 over-expression/interfer?ence vector, and the change of transcription profile were detected by microarray assay and biological information technology analysis. Results MCP-1 over-expression/interference vector MCP-1-pCDH-GFP/MCP-1-LMP shRNAmir1 was con?structed and HUVECs were transfected. According to the microarray analysis we found that there were 18 down-expressed signaling pathways and 7 up-expressed signaling pathways in MCP-1-pCDH-GFP-transfected cells. There were 60 down-expressed signaling pathways and 15 up-expressed signaling pathways in the MCP-1-LMP shRNAmir1 transfected cells. Conclusion Signaling pathways of MCP-1 plays an important role in DVT formation,which may provide us a new way to study molecular mechanism of DVT.
ABSTRACT
Objective To build rat DVT inferior vena cava partial stasis (narrow) model, to detected the expression ofβ2-GP1, VEGF and TF in rat blood, and to investigat the correlation betweenβ2-GP1, VEGF and TF with DVT. Meth?ods SD rats (n=70) are divided into control group (n=10), sham operation group (n=30) and the model group (n=30) ran?domly and DVT model was built by the inferior vena cava partial stasis (narrow) after 2 h, 8 h and 24 h respectively. In each time point, ten rats were taken in each group, inferior vena cava blood were collected whileβ2-GP1, VEGF and TF expres?sion were detected by ELISA. Results In rat experiment, compared with control group, there was no significant change in?expression of β2-GP1, VEGF and TF in sham operation group (P > 0.05). Levels of β2-GP1, VEGF and TF were in?creased at the 2nd hour and 8th hour then peak at the 24th hour which was higher than those in the 24th hour control group and in Sham group and it was also higher than those in the 2nd hour and the 8th hour in model group with statistical signifi?cant difference (P<0.01). Conclusion Based on the above experimental data, in rat DVT formation process, β2-GP1, VEGF and TF may play an important role in promote DVT formation.
ABSTRACT
BACKGROUND: The molecular mechanism and core regulatory network of deep vein thrombosis are not fully clarified yet.OBJECTIVE: To explore the roles of oxidative stress and Rac1/2 in rat deep vein thrombosis.METHODS: Deep vein thrombosis model in SD rats was established by a champing method femoral veins clamping combinedwith fixation of the lower extremity with plaster. The incidence and serious degree of thrombus were observed by dissecting ratfemoral vein at different time points (2.5 and 25 hours after modeling). The model rats were divided into pre-thrombogenesisgroup (2.5 hours after modeling), thrombogenesis group (25 hours after modeling) and non-thrombogenesis group (25 hours aftermodeling). Then total RNA and protein were extracted from the femoral venous wall tissues.RESULTS AND CONCLUSION: Colorimetry results showed that compared with the non-thrombogenesis group, theconcentration of malondiadehyde in rat femoral vein wall tissues of the thrombogenesis group was the highest (P < 0.05), followedby that of the pre-thrombogenesis group (P < 0.05). The concentrations of total superoxide dismutase and glutathione reductasein the thrombogenesis group were the lowest, followed by those in the pre-thrombogenesis group (P < 0.05). The results of genechip hybridization analysis and real-time PCR showed that compared with the non-thrombogenesis group, the expressions ofRac1 and Rac2 in rat femoral vein wall tissues of thrombogenesis group increased the most, followed by that of thepre-thrombogenesis group (P < 0.05). These findings indicate that the up-regulation of malondialdehyde and Rac1/2 as well asthe activity decrease of total superoxide dismutase and glutathione reductase may lead to the formation of deep venousthrombosis.
ABSTRACT
BACKGROUND: At present, the basic underlying molecular mechanism regulating the interactions among venous endothelial cells, platelets, leukocytes, and promoting local deep vein thrombosis microenvironment formation, still remains unclear, and there is no ideal method for early diagnosis of deep vein thrombosis. OBJECTIVE: To study the underlying role of nuclear factor kappa B1 and tissue factor in rats with deep vein thrombosis. METHODS: A total of 67 Sprague-Dawley rats were randomly divided into control group (n=10) and model group (n=57). Deep vein thrombosis model was established by a clamping and sewing method in femoral vein combined with cast fixation. The incidence and serious degree of thrombus were observed by dissecting rat femoral vein in different time points (2.5 and 25 hours after modeling). The model group was further divided into pre-thrombogenesis group (2.5 hours after modeling), thrombogenesis group (25 hours after modeling) and non-thrombogenesis group (25 hours after modeling). Then total RNA was extracted from the localized femoral venous endothelial tissue. The candidate genes, associated inflammation and thrombosis, were screened by a special gene chip. Then the gene expression of nuclear factor kappa B1 and tissue factor was further identified by real-time polymerase chain reaction. RESULTS AND CONCLUSION: Pre-thrombogenesis group had no thrombogenesis, while thrombogenesis group have 23 cases with thrombosis and non-thrombogenesis group have 22 cases without thrombosis. The results of gene chip hybridization analysis and real-time PCR found that the mRNA expression of nuclear factor kappa B1 and tissue factor in rat femoral vein endothelial tissue were significantly up-regulated at 2.5 hours after modeling (pre-thrombogenesis group was higher than control group) (P < 0.05), and continued up-regulating at 25 hours after modeling (thrombogenesis group was higher than the pre-thrombogenesis group, non-thrombogenesis group and control group) (P < 0.05). The results from present study indicate that up-regulating expressions of nuclear factor kappa B1 and tissue factor in local femoral venous endothelial tissue of rat deep vein thrombosis models may play a key role in initiating venous thrombosis.
ABSTRACT
BACKGROUND: At present, the basic molecular etiological mechanism and core regulatory network of deep vein thrombosis (DVT) remains uncertain, and there is not an ideal measure for early diagnosis of DVT. OBJECTIVE: To study the underlying impact of cathepsin L/G in DVT rat model. METHODS: DVT rat models (n = 50) were established by clamping both femoral vein in three different positions within 3 seconds with mosquito forceps and fixing with cast. According to different observation phases and biological situations of the femoral vein thrombosis, model rats were divided into thrombogenesis group, pre-thrombogenesis group and non-thrombogenesis group. An additional 10 normal rats served as control group. Femoral vein was obtained at corresponding time points to exact total RNA. After a gene chip-based screening, the data of gene expression were further dissected by real-time PCR. RESULTS AND CONCLUSUON: Gene chip hybridization analysis results demonstrated that differential expression of cathepsin L/G gene was significant among groups, and the expression was greatest in the thrombogenesis group, followed by pre-thrombogenesis and non-thrombogenesis groups, which was significantly greater than the control group (P < 0.05). Real-time PCR analysis results were consistent with gene chip hybridization analysis results. These indicate that DVT is associated with an increase in expression of cathepsin L/G in local venous vascular wall, and they may be candidate molecular markers for early diagnosis of deep vein thrombosis.
ABSTRACT
BACKGROUND: Deep venous thrombosis (DVT) always occurs after orthopedic surgery. At present, clinical diagnosis of DVT has been lack of an effective measuring means for a long time. Cathepsin may be an effective biological marker of DVT. OBJECTIVE: To study the expression change of cathepsin B and cathepsin C in the rat blood cells before and after DVT and to investigate the feasibility of cathepsin B and cathepsin C as candidate molecular markers for early diagnosis of DVT. METHODS: Totally 100 Sprague Dawley rats were randomly divided into normal control group (n=10) and model group (n=90). Rat traumatic deep vein thrombosis models were established by clamping the femoral vein and fixing the bilateral hind limbs. According to observation time points and the different situations of thrombosis, rat models were assigned to three subgroups: pre-thrombosis, intra-thrombosis, and non-thrombosis. Blood RNA of each group was extracted and reverse transcribed into cDNA. The expression of cathepsin B and cathepsin C in blood cells was detected using real-time fluorescence quantitative PCR. RESULTS AND CONCLUSUON: Expression of cathepsin B and cathepsin C in the blood cells was obviously expressed in the intra-thrombosis subgroup. There was no significant difference in cathepsin B and cathepsin C expression between pre-thrombosis, non-thrombosis groups and normal control group. These findings suggest that cathepsin B and cathepsin C are closely related to DVP and they can be used as the candidate molecular markers for early diagnosis of DVT.
ABSTRACT
BACKGROUND:There is lack of an effective measuring means to diagnose deep venous thrombosis (DVT) in clinic.KLF2 and KLF4 are down-expressed at prethrombotic state,which may be served as predictive molecular markers to diagnose DVT.OBJECTIVE:To explore the feasibility of KLF2 and KLF4 as molecular markers to prediagnose DVT in rats.METHODS:Totally 90 rats were obtained from 100 rats to establish traumatic DVT models and divided into the prethrombotic,thrombosis crest-time and non-thrombosis groups.The remained 10 rats served as control group.Rat blood was collected at each time point,and the expressions of KLF2 and KLF4 were detected by real-time PCR.RESULTS AND CONCLUSION:The KLF2 and KLF4 mRNA expressions in the prethrombotic group and thrombosis crest-time group were lower than that of the control group.However,the KLF2 and KLF4 mRNA expressions in the non-thrombosis group was higher than that of the control group.Therefore,KLF2 and KLF4 may be candidate molecular markers for prediagnosis of DVT in rats.
ABSTRACT
BACKGROUND:Studies in recent years have demonstrated that arginase Ⅰ contribute to the process of numerous cardiovascular diseases,however,most of studies focus on arteries,few regarding venous diseases.OBJECTIVE:To explore the changes of arginase Ⅰ expression in rat traumatic deep venous thrombosis models,and to analyze the possible function of arginase Ⅰ in deep venous thrombosis formation.METHODS:Totally 100 Sprague Dawley rats were randomly divided into the control and model groups.Traumatic deep venous thrombosis models were established by clamping the femoral vein and immobilizing the bilateral hind limbs (hip spica cast fixation),and assigned into initial thrombosis,peak thrombosis and non-thrombosis groups according to different observing time points and pathophysiological situations of thrombosis.Whole blood RNA of each group was extracted,and the change of arginase I expression in blood cells of each group was detected by real-time PCR.RESULTS AND CONCLUSUON:Expression of arginase Ⅰ in the peak thrombosis group was significantly increased compared with other 3 groups (P < 0.01).There were no significances among control,initial thrombosis and non-thrombosis groups (P > 0.05).The finding demonstrated that arginase Ⅰ is closely related to deep vein thrombosis formation.
ABSTRACT
BACKGROUND:The molecular mechanism of traumatic deep vein thrombosis is complex.Numerous studies focus on clinical observation and epidemiology,but its molecular mechanism has not been a new breakthrough.OBJECTIVE:By use of gene array technology,this study was aimed to study the expression changes of matrix metalloproteinases in rat models of traumatic deep vein thrombosis,and to explore the roles of matrix metalloproteinases in traumatic deep venous thrombosis.METHODS:A total of 150 SD rats,SPF grade,of 8-12 weeks old,body weight of 250-300 g,were divided at random into normal control group (n=10) and model group (n=140).Rat traumatic deep venous thrombosis models were set up by clamping the femoral vein and fixing the bilateral hind limbs,and the fixation of hip spica with plaster bandage was conducted in each group.Then rats were divided into 7 subgroups:post-traumatic 0.5 hours,post-traumatic 2.5 hours (initial period of thrombosis),post-traumatic 25 hours (thrombogenesis at thrombotic crest-time),post-traumatic 25 hours non-thrombogenesis at the thrombotic crest-time),post-traumatic 72 hours (thrombus resolution),post-traumatic 72 hours thrombus insolution) and post-traumatic 168 hours (nonthrombosis).At the corresponding phasess,the femoral vein tissues were incised,and total RNA of femoral vein was extracted using Trizol one-step method.Applying Genechip Rat Genome 430 2.0 genechips,the gene expressions in femoral vein were detected in different groups.The rate of traumatic deep venous thrombogenesis and non-thrombogenesis,the rate of thrombi solution and insolution were observed;the expressions of matrix metalloproteinases and tissue inhibitor of metalloproteinases at different time phases was detected by gene array data analysis.RESULTS AND CONCLUSION:Three model rats died and the remaining 147 rats were involved in the final analysis.At the post-traumatic 25 hours,the rate of thrombogenesis was 50.5% and nonthrombogenesis was 49.5%.To the post-traumatic 168 hours,the rate of thrombus solution was 56.7% and thrombus insolution was 43.3%.Both matrix metalloproteinases and tissue inhibitor of metalloproteinases exhibited differential expressions in the course of traumatic deep venous thrombosis.Under the thrombus insolution state,matrix metalloproteinases continued to show a high expression,tissue inhibitor of metalloproteinase expression was down-regulated in the thrombus formation,was significantly inhibited in the thrombus insoluUon process.In the process of traumatic deep vein thrombosis and insolution,matrix metalloproteinase was closely related to traumatic deep vein thrombosis,the matrix metalloproteinase/tissue inhibitor of metalloproteinases are likely to affect the biological state of thrombosis.
ABSTRACT
BACKGROUND: An ideal model of osteonecrosis of the femoral head is beneficial to the study on the cause of disease, pathogenesy and treatment. So far there has not been a coherent method to prepare this model. OBJECTIVE: To establish a rabbit model of osteonecrosis of femoral head induced by microwave heating, and to decide optimum microwave temperature and heating time.DESIGN: Randomized controlled animal trial.SETTING: Animal Experimental Center of Kunming Medical College.MATERIALS: The experiment was performed at the Animal Experimental Center of Kunming Medical College between September 2004 and November 2005. Forty-eight healthy adult New Zealand rabbits, either male or female, were provided by the Animal Experimental Center of Kunming Medical College. The animal procedure was accorded with the ethical standards. GW-92C multi-functional microwave therapy apparatus was the product of Grand World Medical Apparatus (Tianjin) Co., Ltd. METHODS: The microwave antenna was inserted into the rabbit femoral head. Ninety-six femoral heads in forty-eight rabbits were randomly divided into four groups (n =24) according to the microwave temperature and heating time: microwave heating at 50 ℃ for 10 minutes group; 55 ℃ for 10 minutes group; 50 ℃ for 20 minutes group and 60 ℃ for 10 minutes group. The models of osteonecrosis of femoral head were induced by microwave heating using multi-functional microwave therapy apparatus according to the temperature and heating time of grouping. MAIN OUTCOME MEASURES: In each group, two rabbits (four femoral heads) were killed immediately, one, two, four, eight and twelve weeks after operation, respectively. A series of examinations were carried out, including gross observation, X-ray to observe bone trabecular arrangement, cystis degeneration, head collapse or hip joint destruction, MRI to observe the necrotic area, and HE staining to observe the osteonecrosis and bone repair. RESULTS: Marrow tissues partially coagulated in the microwave heating at 50 ℃ for 10 minutes group at the end of the 1st week, and the osteonecrosis returned to normality at the end of the 8th week. In 55 ℃ for 10 minutes group, marrow tissues were completely coagulated at the end of the 1st week, and decreased signal on T1 weighted images and increased signal on T2 images were identified at the end of the 2nd week. In the 4th week, bone repair was found simultaneously when osteonecrosis occurred. At the end of the 12th week, the osteonecrosis continued and the repair stopped, and the femoral head collapse occurred. All femoral heads collapsed at the end of the 8th week in 50 ℃ for 20 minutes group and 60 ℃ for 10 minutes group. CONCLUSION: Microwave heating at 55 ℃ for 10 minutes is the optimal choice to develop a rabbit model of osteonecrosis of the femoral head.
ABSTRACT
Objective To study the effects of emergency operation with 2~5 cm incision to treat fresh closed heel tendon disjunction.Method 23 cases of fresh closed heel tendon disjunction were operated in post-injury 3~12 hours and the incisions were 2~5 cm long. Results None of these cases was complicated with wound infection or cutaneous necrosis.The follow-up was from 6 months to 6 years,averaging 2.2 years;in which,according to the Arner-Lindholm standards,20 were excellent,3 were good.Conclusion A satisfactory effect can be obtained through the smaller incision to treat the fresh closed heel tendon disjunction.
ABSTRACT
0.05)but there was a correlation of IL-1?/IL-10 ratio with thrombus mass (Pearson r=0.87, P=0.01
ABSTRACT
Objective To explore the effect of posterior internal fixation surgery with pedicle screw/rod instrument system in the treatment of thoracic vertebrae fracture-dislocation complicated with a severe thoracic injury. Methods In the 8 cases examined, the range of injury was from T5 to T9, including 1 case of compression fracture-dislocation and 7 cases of burst fracture-dislocation according to Hanley and Eskey’ s classification. According to Frankle’ s classification, 4 were grade A, 3 were grade B, 1 was grade C. All cases were complicated with multiple fractures of bilateral ribs, pulmonary contusion and haemothorax. After the pathogenic condition being stable, reduction and internal fixation were performed with the posterior surgery pedicle screw/rod instrument system. In 5 cases, spinal canal anterolateral decompression was performed homeochronously. The intervals between accidents and operation were 3 to 9 days. Results Operations were performed safely in all patients. The reposition of the alignment and height of vertebraes was good. Postoperative CT scanning showed: in 2 cases, bone block occupied about 10% volume of vertebral canal, and 1 was 50%. The follow-up were 4～36 months, averaging 15 months. No postreduction disposition, internal fixation loosening or breakage occurred. The Frankel’ s classification of neurological function was improved by 1 grade on average. Conclusions After a positive treatment of thoracic injury, it is safe and feasible to treat the fresh thoracic vertebrae fracture-dislocation with the posterior surgery pedicle screw/rod instrument system, and the curative effects are satisfied.