Objective To establish human lung adenocarcinoma multidrug resistance cell lines in vitro,observe their biological characteristics,and investigate the mRNA expressions of DNA pol?,mdr 1,mrp1,GST-?,lrp and topo Ⅱ genes.Methods Paclitaxel-resistant cell lines(A549/TXL20) were established in vitro by exposure to stepwise increased concentrations of the drug in a cell culture medium.Biological morphology and cell cycles were analyzed by morphometry and flow cytometry.The chemoresistance indexes of cells were measured by methyl tetrazolium assay.Evaluation of growth and in vitro drug sensitivity were performed.RT-PCR was employed to analyze the mRNA expressions of the DNA pol?,mdr 1,mrp1,GST-?,lrp,and topo Ⅱ genes.Results ① Compared with parent cells,the resistant sublines had a lower confluent density.They were smaller and mixed with giant cells in different sizes and with different numbers of nucleoli,and the growth property of A549/TXL20 did not change significantly compared with A549 cell lines.② The resistant cells,A549/TXL20,were 19.3 times more resistant to paclitaxel and 67.4 times more resistant to cisplatin than the parent cells,and also demonstrated cross-resistance to mitomycin,vinblastine,and 5-fluouracil(5-FU). ③ Compared with the A549 celllines,an unreasonably higher level of drug resistance and lower drug concentration was detected in A549/TXL20 cells after exposure to the drug in the culture medium.④ The mRNA expression level of DNA pol?,mdr1,GST-?,mrp1 andlrp genes in A549/TXL20 cells was significantly higher than that in A549 cell lines(P

The recombinant plasmid of mChlL-18 prokaryotic expression was transformed into E.coli BL21(DE3) strain and then induced by IPTG at 37℃.After crushed and washed,the expressing inclusion bodies were thoroughly denatured with 6 mol/L guanidine hydrochloride.Then according to experiment design,the effects of rChlL-18 protein refolding yield at different densities were investigated by the systems of artificial chapercne at different densities.Experiment results indicate that there is a optimal condition on assiting rChIL-18 protein by using the artificial chaperone technique.The optimal condition can improve the refolding yield of rChIL-18 protein,and then the expressed product of fusion chicken IL-18 gene in E.coli has a relativity high bioactivity.

This paper is to study the clinical characteristics of Brugada syndrome (BrS) in Chinese by analyzing clinical and ECG data of BrS patients. Data were included by computerized and manual research, and was analyzed by 2 doctors alone. The data of repetition and of non-Chinese were rejected. Forty-nine BrS patients were included (45 males and 4 females). Main manifestations included sudden death in 29 and syncope in 27 patients. Malignant ventricular arrhythmia (MVA) occurred in 14 of 17 patients with family history of sudden death or syncope and in 15 out of 32 ones without family history. Occurrence of MVA in 11 of 14 patients was within the period from 7pm to 7am. ECG revealed that sloped ST segment elevation appeared mainly in V1, V2 leads and coved ST segment elevation appeared mainly in V3 lead. BrS is not rare in Chinese people,and its clinical characteristics is similar to that overseas data.

Objective To summarize our preliminary experience on diagnosis and treatment of aorta pseudoaneurysms due to deceleration injury. Methods Retrospective analysis was made on 8 cases of aorta pseudoaneurysms due to deceleration injury regarding its clinical findings, imagines and surgical operations or endografting treatment. Results Diagnosis was confirmed by imaging examinations. Among two cases who refused a surgery one died and the other lost follow-up after discharge. Surgery was successful in the remaining six cases including two cases treated by open surgery and four cases by intervensive endografting. Conclusions Aorta pseudoaneurysms due to deceleration injury can be correctly diagnosed by imaging examinations. Aorta pseudoaneurysms should be treated timely since spontaneous cure is almost impossible. Traditional surgical operations is effective, however, endografting is a relatively safe, less traumatic procedure.

Objective To evaluate therapeutic results of radiofrequency endovenous obliteration (RFO) for the treatment of varicose veins of the lower limbs. Methods Fifty six cases (56 limbs) of primary greater saphenous vein tributary varicose veins were randomly assigned to RFO group (n=28) and conventional stripping operation group (n=28). In RFO group, the wall of the greater saphenous vein was treated at 85℃ with the catheter to occlude the whole length of the vein. The other 28 cases underwent stripping procedure. The scattered superficial varicose veins in calf in both groups were managed by phlebectomy. The number of surgical incision, postoperative pain, average hospital days and the short-term results were compared. Results Patients in RFO group have less surgical incisions and less postoperative pain, without subcutaneous hematoma. The average hospital stay was 2.5?1.00 days in RFO group compared to 4.14?0.85 days in stripping operation group. Conclusions RFO effectively obliterates the whole length of the great saphenous vein and is of less trauma,faster recovery, and less scars.

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.

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.

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.

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.

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.