Objective To establish and apply the multiple probe RT-PCR analysis for H5N1 pathogen detection methods. Methods Used the gene sequences published in GenBank,which were designed and synthesized two pairs of specific primers (P1/P2,P3/P4)accorded the conserved regions.Through the optimization of amplification conditions to establish the rapid detection of two viruses duplex PCR method.Results The suitable primer concentration:P1/P2 was 0.32μmol/L,P3/P4 was 0.96μmol/L.The amount of the minimum virus nucleic acid was 2 ng/μl by the double detect,H5N1 pathogen mixed were occurred specific bands in the corresponding location,and other viruses and blank control strips had not bands.Conclu-sion A variety of probes for RT-PCR analysis H5N1 pathogen detection methods in a reasonable application of primer con-centration under has good sensitivity and specificity,and it is worth the inspection application.

Objective To investigate the effection of Shenfu Injection (Ginseng-radix aconitum Injection)to the blood coagulation system and its prognosis in the patient of multiple trauma complicated with shock.Methods We prospectively studied 90 patients of multiple trauma complicated with shock,who came from the emergency center of the Second Affiliated Hospital of Medicine' s college of Zhe-Jiang University and emergency department of Lishui People' s Hospital from February 2007 to December 2011,and excluded those suffered with the dysfunction of coagulation system and uncontrolled ongoing bleeding before trauma.And then these eligible patients were randomly divided into two groups:treatment group ( 45 cases ) and control group (45 cases),the control group were received the routine treatment,the treatment group were received Shenfu Injection in early stage based on the routine treatment.The two groups were measured the platelet count (PLT),prothrombin time (PT),activated partial thromboplastin time (APTT),thrombin time (TT),fibrinogen (FIB) before treatment and 3,7 days after treatment.Results The differences of the PLT,PT,APTT,TT,FIB between the treatment group and control group at 3,7 d after the treatment was statistically significant (P ＜ 0.05 ). Conclusions Shenfu Injection has positive regulation to the coagulation system in the patient of multiple trauma complicated with shock.

Objective To investigate the safety and clinical efficacy of catheter-directed thrombolysis(CDT) with prolonged infusion of low dose urokinasefor treatment of acute iliac-femoral vein thrombosis.Methods From January 2005 to March 2011,63 patients of unilateral acute iliac-femoral vein thrombosis were treated by CDT and followed up for more than 12 months.The complications during CDT,thrombus clearance ratio,time for CDT,dose of urokinase,degree of limb swelling and clinical follow-up data were retrospectively reviewed.The Fisher exact test was used for enumeration of data.The measurements of data were tested with the one-way analysis of variance and two-two comparison LSD test.Thrombus clearance ratio in different time frame was tested by repeated measurement of data and analysis of variance.Results During the CDT,no symptomatic pulmonary embolism (PE) and significant bleeding happened.Ten (15.9％) patients had complications of minor bleeding,including 3 patients with blood oozing along the edge of vascular sheath,3 patients with subcutaneous ecchymosis or hematoma around the puncture site,3 patients with hematuria and 1 patient with gums bleeding.In four (6.3％) patients,complications were caused by catheter placement,including 3 patients with soft tissue inflammation around puncture site and 1 patient with secondary thrombosis surrounding the vascular sheath.Fifty three patients (84.1％) achieved thrombolytic degree Ⅲ and Ⅱ at 4th to 8th day during CDT.Thrombus clearance ratio was higher in CDT with urokinase 500 000 U/d and 750 000 U/d than 250 000 U/d[(91.2 ± 10.1)％ vs (75.9±20.1)％,(91.3 ± 12.2)％ vs (75.9±20.1)％,all P ＜0.05].Thrombus clearance ratio showed no significant difference between CDT with urokinase 500 000 U/d and 750 000 U/d [(91.2 ±10.1) ％ vs (91.3 ± 12.2) ％,P ＞0.05].There was no significant difference between CDT with urokinase 500 000 U/d and 750 000 U/d in perfusion thrombolytic time to reach thrombolytic degree Ⅲ [(7.1 ± 1.0)vs (6.2±1.3)d,P＞0.05]and Ⅱ[(6.4±1.0) vs (6.0±0.8)d,P＞0.05].Thrombus clearance ratio increased along with an increase in thrombolytic time for CDT (P ＜ 0.05).After 24 hours of CDT,58 (92.1％) patients showed reduction of soft tissues tension.After 48 hours of CDT,affected limb circumference decreased significantly compared with the preoperative measurement [thigh (54.25 ±5.79) cm vs (56.46±5.91) cm; leg(44.05 ±5.18) cm vs (45.68 ±5.16) cm,all P＜0.05].At the time of discharge,there was no significant difference between affected limb circumference and normal side [thigh (49.00±4.67) cmvs (48.38 ±4.68) cm; leg(38.41 ±4.15) cm vs (37.73 ±3.92)cm,all P ＜ 0.05].The patency rate of iliac venous stent was 91.1％ (41/45) after 6 months.Doppler ultrasound showed regurgitation of femoral venous valve in 11 patients after 12 months.Conclusions CDT with prolonged infusion of low dose urokinase is a safe,highly effective method for the treatment of acute iliacfemoral venous thrombosis.

ObjectiveTo investigate prospectively the feasibility and clinical value of catheterization via the ipsilateral great saphenous vein in catheter-directed thrombolysis (CDT) for acute iliofemoral deep vein thrombosis (IFVT) by a comparative study.MethodsThe prospective study included 93 cases of IFVT proved by venography.All patients were divided into three groups randomly.In group A,31 patients received CDT via the ipsilateral great saphenous vein.In group B,27 patients received CDT via the ipsilateral popliteal vein.In group C,35 patients received anterograde thrombolysis via an ipsilateral dorsalis pedis vein.Urokinase was adopted as the thrombolytic agent in all cases.The assessment of the curative effect include therapeutic effective rate,rate of edema reduction and venous patency which were observed according to the clinical symptoms and the follow-up venograms obtained 5 days after thrombolysis.The time and comfort scores of procedures was recorded and compared between group A and B using two independent samples t test.The rate of edema reduction and venous patency were assessed using analysis of variance (LSD method).Therapeutic effective rate and complication rate were assessed using Chi-square test.Results The total effective rate of the three groups were 90.3％ (28/31),92.6％ (25/27) and 68.6％ (24/35) respectively.The limbs edema reduction rate were (83.5 ±21.1)％,(82.4 ±20.1)％,and(67.0±23.3)％ respectively(F=6.059,P = 0.003 ).The venous patency rate after thrombolysis were (61.2 ± 20.2) ％,(55.7 ± 20.5 ) ％,and (44.2 ±23.6)％ respectively.There was no significant difference between group A and B in therapeutic effective rate( x2 =0.09,P =0.759),rate of edema reduction( P =0.822 ) and venous patency ( P =0.343 ).There was a significant difference statistically in therapeutic effective rate(x2 =4.65,P =0.031 ),rate of edema reduction (P = 0.002) and venous patency (P = 0.002) between group A and C.Compared with group A and B,the procedure time [group A (8.3 ±3.1) min,group B (16.3 ±3.5) min,t =9.379,P ＜0.05],comfort scores during treatment [ group A (2.2 ± 1.2),group B (5.0 ± 1.4 ),t = 8.129,P ＜ 0.05 ] had statistical significant difference.The CDT-asscciated complications in group A were less than group B significantly(3 cases in group A,11 cases in group B,x2 =7.60 P ＜0.05).ConclusionsCatheterizationvia the great saphenous vein in CDT therapy for acute IFVT is feasible and effective.It is easily operable with less complications.

Objective To evaluate the value of interventional therapy in treatment of iliac compression syndrome (ICS) and subsequent venous thrombosis. Methods Examined by DSA, 125 cases were diagnosed of iliac vein compression and subsequent thrombosis. In 39 cases of ICS ( group 1 ), left: right = 4.6: 1. In 86 cases of ICS complicated with subsequent thrombosis (group 2), left: right = 4.7: 1. The patients of iliac vein compression and compression-related iliac vein stenosis or occlusion without fresh thrombus were treated by percutaneous transluminal angioplasty (PTA) and self-expandable stenting. In those cases with fresh thrombosis the inferior vena cava filter were inserted before thrombosis suction, mechanical thrombus ablation, PTA, stenting and transcatheter thrombolysis. The Chi-square test for comparison of proportions was used to test statistical significance. Results In 39 cases of ICS, 38 cases were treated by PTA and stenting. In 86 cases of deep vein thrombosis complicated with ICS, 83 cases were treated by various interventional therapy. There was no significant difference in the efficiency of intraluminal treatment between the two groups at discharge (97.4% and 96.5%, X2 =0.000,P >0.05) and at 6 months follow-up(96.3% and 90.2%, X2 = 0.266, P > 0.05 ), the difference in excellent-good rate of the two groups was significant at discharge (94.9% and 79.1%, X2=3.879, P <0.05) and at 6 months follow-up (92.6% and 68.6% ,X2 =4.441,P <0.05). Conclusions Interventioual treatment for ICS and secondary thrombosis is safe and effective.

Objective To assesse prognostic factors regarding long-term primary patency for patients who underwent intra-arterial thrombolysis and/or adjuvant endovascular techniques due to acute lower limb ischemia. Methods Consecutive patients with ALI of the lower extremities treated via interventional methods between January 2005 and June 2010 were identified and reviewed ( exclude patient suffered from aortic dissection involved artery of lower extremity or trauma). Analyze the potential variables with univariable analysis and only factors associated with long-term primary patency with a P value less than 0.1 in univariable analysis were introduced into the Cox regression mode.Total long-term primary patency and grouped primary patency were assessed using Kaplan-Meier estimation.Results The analyzed dataset included 107 limbs treated in 101 patients presenting with ALI (class Ⅰ 15,class Ⅱ A 36,class Ⅱ B to Ⅲ 56,according to Rutherford classification ).Eight nine limbs were enrolled in follow-up.The mean followup was 34 months ( range:1 to 53 months).Primary patency at 12,24 and 36 months was 87％,68％ and 55％,respectively.Multivariable analyses identified patients presenting with diabetes mellitus ( P =0.00),PAOD ( P ＜ 0.02 ) and thrombolysis time ( P ＜ 0.02 ) were associated with primary patency.Compare the patency rate of patients with different thrombolysis time,the results showed that the patency rate of the patients thrombolysis time less than 4 d was higher than those more than 4 c. Conclusions lnterventional therapy remains an effective treatment option for patients presenting with lower extremity ALI.Diabetes mellitus and PAOD negatively affect the rates of limb primary patency. Thrombolysis should be limited to ＜4 days.

ObjectiveTo investigate the anatomical distribution of acute deep venous thrombosis (DVT) of the lower extremity,and compare different therapeutic methods including anticoagulation alone,thrombolysis through dorsal vein and interventional therapy.MethodsThe clinical data,venography and therapies of 204 acute DVT patients were retrospectively studied According to the distribution,DVT were classified into three types including peripheral,central and mixed types.According to the difference of the therapeutic method,each type of DVT was divided into three groups,Group A (37 patients) anticoagulation alone:Group B(55 patients) thrombolysis through dorsal vein:and Group C( 112 patients) interventional therapy.The results of different kind of treatment method in each type of DVT were evaluated before the patients were discharged and the Chi-square test was used for statistical analysis.Results There were 132 patients with DVT in the left lower extremity,62 in right lower extremity,and 10 in both extremities.The complication of pulmonary embolism (PE) occurred in 4,5 and 2 cases respectively,and the morbidity was 3.0％,8.1％ and 20.0％ ( x2 = 6.494,P = 0.039 ) respectively.There was significant statistical difference among them.There were 23 cases of peripheral type of DVT,48 central type and 133 mixed type.The complication of PE were observed in 2,5 and 4 cases respectively in each type.The morbidity was 8.7％,10.4％ and 3.0％ respectively ( x2 = 4.350,P = 0.114 ).There were no statistical significance among them.In the 23 cases of peripheral type DVTs,2 of 5 in group A and 11 of 18 in group B had excellent therapeutic response.In the 48 cases of central type of DVTs,1 of 10 in group A,2 of 5 in in group B and 26 of 33 in group C had excellent therapeutic response.There were statistically significant differences among groups A,B and C ( x2 = 16.157,P =0.000).In the 133 cases of mixed type DVTs,1 of 22 in group A,10 of 32 in group B and 65 of 79 in group C had excellent therapeutic response.There were statistically significant differences among group A,B and C ( 1,10,65 cases,x2 = 53.993,P =0.000).ConclusionsThe incidence of acute DVT involving the left lower extremity was higher than that involving the right one,and the majority of cases was of the mixed type.The treatment of choice for the central and mixed types was interventional therapy.Analysis of anatomical distribution of deep venous thrombosis can guide treatment planning.

Objective To investigate the short-term efficacy and security of combined treatment of iodine-125 seeds and transeathether arterial chemoembolization in liver neoplasms. Methods Transcathether arterial chemoembolization underwent in experimental group (28 cases) with liver neoplasm. The treatment plan was formulated with treatment planning system and a median of 25 seeds per patient (range, 15--40 seeds) were implanted under CT or B ultrasound guidance in 2 weeks after the procedure. Transcathether arterial ehemoembolizafion underwent after the implanted regularity. Blood routine and liver function were detected before and after the procedure. X ray check and abdomen CT scan were performed each 2 months. Control group (32 cases) were treated with transcathether arterial chemoembolization alone. Analysis of variance and Chi-square test were used for statistics. Results All seeds were released to the target places successfully and no seed was found to be lost or migrated in experimental group. Transient elevation of the serum ALT and AST but recovered in 2 week. WBC, Hb, IgA and IgG were showed no significant changes. The severe complication was not found in those eases. The responsive rate of tumor was 75.0% (21/28), 37.5% (12/32) in experimental group and control group, respectively(X2 = 8.485,P = 0.004). The survival rate of 6 months was 92.9% (26/28), 75.0% (24/32) in experimental group and control group, respectively(X2=2.263,P=0.132). The surviral rate of 12 months was 72.0% (18/25), 43.3% (13/30) in experimental group and control group, respectively (X2 = 4.556, P=0.033). Conclusion It is simple, feasible, safe and short-termly effective for liver neoplasms in treatment combined iodine-125 seeds implantation with transcathether arterial chemoembolization.

Objective To demonstrate lower limb angiography results of peripheral arterial disease (PAD) in diabetics and evaluate one-year curative effect after interventional therapy. Methods Lower limb angiography results and the efficiency of interventional therapy for 44 limbs with PAD in 38 diabetics were retrospectively analyzed. Post-treatment clinical manifestations, signs and ankle-brachial-index (ABI) at 1 week, 1 month, 3 months, 6 months and 12 months were compared with those before treatment. Clinical evaluation was divided into four grades: apparent, effective, ineffective and deterioration. Efficiency =(apparent + effective ) / total cases ÷ 100%. ABI was compared using analysis of variance. Results Lower limb angiography revealed multi-branch lesions, with multi-segmental stenoses or obstructions.Lesions involved both above- and below-the-knee arteries in 25 limbs (56. 8% ), only above-the-knee arteries in 3 limbs (6. 8% ) and only below-the-knee arteries in 16 limbs (36. 4% ). In the limbs only with below-the-knee arterial lesions, the involved artery branches were one in one limb (2. 3% ), two in six limbs ( 13. 6% ) and three in nine limbs ( 20. 5% ), respectively. The technical success rate of percutaneous transluminal angioplasty (PTA) was 91.4% ( 53/58 ) for diseased below-the-knee arteries. Among the 12 cases with foot and ankle ulcers, ulcers healed within 3 months in 9 cases; however, the other three cases suffered below-the-ankle (in one case) or below-the-knee amputation (in two cases) within 6 months. Four cases with gangrene suffered below-the-knee amputation within one month after PTA. The amputation rate was 15. 9% (7/44). At 1 week, 1, 3, 6 and 12 months after PTA, the effective rates were 79. 6%(35/44), 83.3% (30/36), 85.7% (24/28), 85.0% (17/20) and 81.3% ( 13/16), respectively; ABI values were 0. 86 ± 0. 10, 0. 85 ± 0. 10, 0. 83 ± 0. 11, 0. 79 ± 0. 12 and 0. 75 ± 0. 12, respectively.Compared with pre-PTA ABI value (0. 53 ±0. 20), post-PTA ABI value was significant higher (F=35.79,P ＜ 0. 05 ). However, ABI value began to decline from the 6th month after PTA. Conclusions In diabetics, PAD always involves both above- and below-the-knee arteries. PTA is a feasible and effective revascularization therapy, which could improve the clinical signs and symptoms of lower limb ischemia in diabetics with PAD. The clinical effect is satisfactory during short-term follow-up.

Objective To find out the best method for elimination of uranium contamination in rat tissues as low as possible by removal of shrapnel fragments. Methods Experimental rats were divided into six groups: route group, decontamination before surgery group, decontamination in incision group, changing surgical appliances group, removing tissues around group, and comprehensive method group. Uranium concentrations in tissues and fluids in all groups were measured at 7, 14, and 21 d after operation. The efficiency of decontamination by different methods was compared. Results The highest uranium concentration in tissues was found in the route group, but the lowest in the comprehensive method group, and the second lowest in removing tissues around group. Conclusion The comprehensive method is the best one in all of the surgical removal methods. The soft tissues around DU shrapnels should be removed if they are not critical organs.