Objective To evaluate catheter-directed thrombolysis via the popliteal vein in the treatment of the lower limb acute deep venous thrombosis.Methods From July 2009 to October 2010,32patients of the lower limb acute deep venous thrombosis including 3 patients with concurrent pulmonary embolism underwent uhrasound-guided catheter-directed thrombolysis via the popliteal vein.The thrombolytic catheter was inserted into thrombus,through which urokinase was infused at a dosage of 100 × 104U/d.The venous patency score and the rate of patency improvement were observed by venograms before and after therapy.Results In every patient,the lower limb swell and pulomonary symptoms relieved.The circumferences between affected and normal thigh before and after the thrombolysis were(5.4 ± 1.4)cm and (1.7 ± 1.3)cm(t =9.92,P ＜0.01).The circumferences between affected and normal leg before and after the thrombolysis were(4.1 ± 1.5)cm and(1.5 ±0.7)cm(t =7.65,P ＜0.01).The venous patency score before and after the thrombolysis were(15 ± 4)and(4 ± 3)(t =7.12,P ＜ 0.01).The mean rate of venous patency was 88.21％.In the 3 patients with pulmonary embolism,thrombus was complete dissolved in 1 case and partial dissolved in 2 cases.No major complications occurred in all these patients.29 patients were followed up for 3-12 months.There were no thrombosis relapsed.Conclusions Catheter-directed thrombolysis via the popliteal vein with urokinase for acute lower limb deep venous thrombosis is safe and effective.

In order to investigate the effect of sequence-specific small interfering RNA on suppressing cyclin D1 expression and proliferation and cell cycle and expression of G1 phase regulators of fibroblasts derived from keloid, the plasmid expression vector of siRNA targeted against cyclin D1 was constructed and transfected into fibroblasts with LipofectamineTM 2000. The changes of cyclin D1 expression were detected by fluorescent quantitative PCR(FQ-PCR), semi-quantitative RT-PCR. The effect of sequence- specific small interfering RNA in suppressing the proliferation of fibroblasts was detected by MTT assay. Flow cytometry were used for evaluation ofceU cycle. The expression of cyclin D1, CDK4, pRb and P16 was detected by immunohistochemical method. The results showed that: (1) The sequence- specific siRNA effectively suppressed cyclin D1 expression at both mRNA levels with inhibition rate of 63.68% and 92.83% (P<0.01). (2) Significantly inhibited the proliferation of fibroblasts, and changed cell cycle in percentage of G0/G1 phase cells was increased compared with the controls groups in fibroblasts(P < 0.05). (3) 72 h after transfection, the expression of cyclin D1, CDK4 and pRb decreased, and the expression of P16 increased. It can be concluded that the plasmid expression vector for the RNAi against cyclin D1 constructed in the study can effectively and specifically suppress cyclin D1 expression, and progression of G1/S is effected by G1 phase related regulatory protein, and suppresses proliferation of fibroblasts derived fiom keloid.

Objective To evaluated the role of wt-P53 protein in telomerase regulation in keloid fibroblasts(KFBs). Methods The fibroblasts were derived from human keloid tissue which was proved by pathological diagnosis. KFBs were divided into 2 groups, the transfection group and the untransfection group. wt-p53 gene was transfected into the fibroblasts by adenovirus vectors in the transfection group. The KFBs untransfected with wt-p53 gene served as control (untransfection group). After 48 hours of transfection, the expression of wt-P53 protein was analyzed by both Western blotting and immunofluorescence method, respectively. The telomerase activity was evaluated by TRAP-ELISA after 1-7 days of transfection.Results All the KFBs from 2 groups expressed wt-P53 protein. But the expression level of wt-P53 protein in the transfection group was significantly higher than that in the untransfection group. At the same time of high expression of wt-P53 protein, the telomerase activity of KFBs in transfection group was significantly lower than that in the untransfection group( P＜0.05). Conclusion High level expression of wt-P53 protein can transiently inhibit the telomerase activity of KFBs.

Objective To investigate the risk factors of acute kidney injury(AKI) after intracoronary stent implantation in order to provide the basis for clinical prophylaxis and treatment.Methods Retrospectively analyzed 626 consecutive patients who underwent isolated intracoronary stent implantation in our institution from January 2007 to July 2011.Multivariate logistic regression model was constructed to identify the risk factors for the development of AKI defined as a serum creatinine (SCr) 130 to 199 μ mol/L or estimated creatinine clearance(Ccr) 30 to 60 ml/min per 1.73 m2.Results Ninety-three patients of 626 (14.9％) underwent isolated intracoronary stent implantation developed AKI.The results of the multivariate forward stepwise logistic regression analysis found that risk factors for the development of AKI following isolated intra-coronary stent implantation was associated with age (OR =1.570,95％ CI 1.308-1.885),ejection fraction (EF) ≤ 30％(OR =11.526,95％ CI 2.452-54.177),hypotension during perioperative and postoperation (OR =11.074,95％ CI 2.439-50.282),operation duration(OR =1.032,95％ CI 1.012-1.051),sex (OR =0.010,95％ CI 0.001-0.086),NYHA class Ⅲ & Ⅳ (OR =0.209,95％ CI 0.059-0.737),peripheral vascular disease (OR =0.528,95％ CI 0.286-0.973),chronic obstructive pulmonary diseases (OR =0.546,95％ CI 0.304-0.982),preoperation Cr (OR=1.418,95％CI 1.216-1.654) (and all P＜0.05).Conclusion AKI is the common complications after intracoronary stent implantation,especially age,EF ≤ 30％,hypotension during perioperative and postoperation,operation duration are independent risk factors.

Objective To observe the expression of retinol-binding protein 4 (RBP4) and phosphatidylinositol 3 kinase (PI3K) in insulin resistant (IR) rats and the role of pioglitazone intervention. MethodsThirty-five male Wistar rats in SPF level were randomly divided into control fed with normal diet (n= 11 ) and IR model fed with high fat sucrose diet (HFSD) (n= 24). The IR rats were then randomly divided into two subgroups, namely IR fed with HFSD(n = 12) and pioglitazone intervention (n= 12) given a daily dose of 20 mg · kg 1 · d-1 pioglitazone and HFSD (IR +Pio group )for 8 weeks.All rats were killed after 16 weeks and the levels of serum TG, high density lipoprotein (HDL-C), low density lipoprotein (LDL-C), fasting blood-glucose (FBG), fasting serum insulin (FIns) and insulin resistance index (HOME-1R) were measured.Enzyme-linked immunosorbent assay (ELISA) and RT-PCR were employed to detect the level of serum RBP4 and the mRNA expression of RBP4 in the epididymis adipose tissues, respectively. Immunohistochemistry was used to detect the protein expression of PI3K in skeletal muscles while UV spectrophotometer was employed to measure level of serum AOPP. The ratio of visceral fat weight of mesentery, epididymis and peritoneum in abdominal cavity to body weight (BW) was calculated. Results(1) the level of BW, TG, LDL-C, FINS and the ratio of visceral fat weight to BW were higher in IR group than in control group, but HDL-C decreased significantly. After the intervention of pioglitazone, the level of BW,TG, LDL-C, FINS, and the ratio of visceral fat weight to BW in IR+Pio group were lower and HDL-C increased significantly than those in IR group.(2) The level of RBP4 from serum and epididymis adipose and serum AOPP were higher in IR group than in control and lower significantly in IR+Pio group than in IR group (all P＜0.05). (3) The expression of PI3K in skeletal muscles were lower in IR group than in NC group, and increased after the invention of pioglitazone. (4) FIns, the ratio of visceral fat weight to BW and LDL-C were positively correlated with RBP4 while HDL-C and PI3K negatively correlated with RBP4. Conclusions(1) The increased RBP4 can lead to metabolic disturbances and oxidative stress in IR rats.(2) RBP4 may decrease the insulin sensibility by weakening insulin signal transduction. (3) Pioglitazone can ameliorate insulin resistance by decreasing the level of RBP4 and serum AOPP and increasing the level of PI3K in skeletal muscles of IR rats.

Objective To observe the effect of intra-articular injection of sodium hyaluronate (Na-HA) on mRNA expression of inducible nitric oxide synthase (iNOS) in cartilage of osteoarthritis (OA). Methods Sixteen white rabbits were underwent unilateral anterior cruciate ligament transection and were divided into 2 groups randomly 5 weeks after transection. Experimental group rabbits received 0.3 ml of intra-articular 1% Na-HA injection once a week. Animals in control group were treated with the same conditions using normal saline. All animals were sacrificed 10 weeks following surgery. The aritcular cartilage degeneration of medial femoral condyle was evaluated at mascroscopically and microscopically. The mRNA expression of iNOS in cartilages was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Results Cartilage degradation in experimental group was significantly less severe than that in control group. No significant difference of iNOS mRNA expression in cartilage was found between the experimental group and the control group. Conclusion Na-HA can significantly reduce the severity of cartilage degradation of OA. Na-HA dose not show definite effect on iNOS expression in cartilage during early stage of OA.

AIM To study anti CMV effect of S ODNs complementary to the initial code region of major immediate early (MIE) mRNA, and to the intron1/exon2 boundary of MIE mRNA precursor. METHODS To evaluate the anti CMV effect by determining viral antigen on infected 2BS cells by in situ ELISA. RESULTS Both of 2 AS S ODNs and their sense sequence control showed anti CMV effect. The medium effective concentration (EC 50 ) were 4 53, 10 2, 26 2 and 30 1 ?mol?L -1 . The secretion of CMV antigens were delayed by 3 d～4 d by 8 0 ?mol?L -1 AS 1. It showed increased effect by administrating earlier postinfection, by supplementing the S ODN at intervals, by conbinating with ganciclovir, and by packaging with liposome. A slight cytotoxicity was observed at the concentration of 64 0 ?mol?L -1 . Northern blot analysis indicated that the MIE mRNA decreased after the treatment of AS 1. It suggests that disintegration of the mRNA in the hybridized duplex by activated RNase H played an important role as the mechanism of specific action, and "time and effect" analysis suggested that interference of viral adsorbtion and penetration may be the important mechanism of nonspecificity. CONCLUSION AS S ODNs targeted to MIE gene are effective anti CMV agents which can be developed as a new type of chemotherapy drugs against CMV infection.

Objective To study the expression of GRHL-3 in diffuse large B-cell lymphoma (DLBCL) tissues and its clinical significance.Methods One hundred and sixty-eight pathology paraffin-embedded diffuse large B-cell lymphomas tissues were collected from January 2006 to September 2011.Immunohistochemistry was used to detect the expression of GRHL-3 protein in the above tissues.Results The positive expression rate of GRHL-3 protein in the GCB type tissues was higher than that in the non-GCB type tissues [84.87 ％(101/119) vs 14.29 ％ (7/49), P ＜ 0.01].Further analysis indicated that in the non-GCB type tissues,the positive expression rate of GRHL-3 in the latter stage group was significantly higher than that in the early stage group [90.00 ％ (63/70) vs 77.56 ％ (38/49), P ＜ 0.01].The positive expression rate of GRHL-3 in the lactatede hydrogenase increased group was significantly higher than that in the normal lactated hydrogenase [91.67 ％ (77/84) vs 68.57 ％ (24/35), P ＜ 0.01].The positive expression rate of GRHL-3 in the extranodal involvement status ≥ 2 group was significantly higher than that in the extranodal involvement status 0-1 group [96.29 ％ (26/27) vs 81.52 ％ (75/92), P ＜ 0.05].The positive expression rate of GRHL-3 in the IPI score 4-5 group was significantly higher than that in the IPI score 0-1 group [91.30 ％ (65/69) vs 66.67 ％ (18/27), P ＜ 0.01] and IPI score 2-3 group [91.30 ％ (65/69) vs 79.96 ％ (18/23), P ＜ 0.05].However, the expression of GRHL-3 had no correlation with the gender, age, and performance status of DLBCL.Conclusion The positive expression rate of GRHL-3 protein in the GCB type tissues is higher than that in the non-GCB type tissues.The positive expression rate of GRHL-3 in the DLBCL is correlated with the Ann Arbor stage, lactate dehydrogenase, extranodal involvement status and IPI score.

A matrix solid phase dispersion extraction_dispersed liquid phase microextraction_gas chromatography_mass spectrometric method has been developed for the determination of three pyrethroids ( tetramethrin, permethrin and deltamethrin) in soil. The optimal conditions for the analysis were as follows. About 0. 5 g soil and 1. 5 g C18 solid phase extraction powder were grinding for 5 min. The mixture was eluted with 10 mL of acetone. The eluent was concentrated to 0. 4 mL, and then mixed with 20 μL of tetrachloromethane and 5. 0 mL of ultrapure water to form a homogeneous cloudy solution. The emulsion was broken by centrifugation. About 1 μL of sediment was injected and analyzed directly by GC_MS. Good linearities for three pyrethroids were ranged from 5 to 200 μg/kg (r2≥0. 9989), and recoveries at three spiked levels were ranged from 86 . 5% to 108 . 0% with RSDs less than 7 . 8%. The LODs of three pyrethroids were 1. 00-1. 48 μg/kg. This method can meet the determination of trace pyrethroids in soil.

Objective To investigate the relationship between aggrecan and YKL-40 in knee articular cartilage of Sprague-Daw-lay(SD) rats with osteoarthritis (OA) .Methods Fifty-six healthy SD rats were randomly divided into 7 groups ,8 cases per group . The one side of knee joint was randomly selected for performing the anterior cruciate ligment transection (ACLT) and establishing the OA model .The rats in one group were randomly killed on the day of operation and at postoperative 0 ,2 ,4 ,8 ,12 ,16 ,20 weeks . The femoral condyle cartilage samples at different time periods in the operated side were collected for conducting safranin O /fast green staining and HE staining .Meanwhile ,the OA pathological grade was made out according to the modified Mankin scale .The expression of aggrecan and YKL-40 in the cartilage with different stages of OA were analyzed by the immunohistochemistry meth-od ,and the status of expression were measured by average optical density (AOD) .The correlation between aggrecan and YKL-40 was analyzed .Results With the aggravation of OA ,the expression of aggrecan was gradually reduced and the expression of YKL-40 was gradually increased .The differences during the early ,middle and late phases of OA had statistical significance (P<0 .05) . The expression of aggrecan was negatively correlated with the expression of YKL-40(P<0 .05) .Conclusion The level of aggrecan is gradually reduced with the aggravation of OA .Aggrecan is negatively correlated with the YKL-40 level ,which may reflect the dedifferentiation degree of joint chondrocyte to some extent .