Objective To investigate the role of TLR4/NF-kB signaling pathway in inhibited invasion ability of pancreatic cancer cells caused by triptolide (TP).Methods PANC1 cells were divided into parental cells group,TP group,lipopolysaccharide (LPS) group and TP + LPS group.50 ng/ml of TP was added in culture medium in TP group,and 1 μg/ml of LPS was added in culture medium in LPS group,while 50 ng/ml of TP was pretreated for 2 h and 1 μg/ml of LPS was added in culture medium in TP + LPS group.All the ceils were cultured for 24 h.The TLR4 and matrix metalloproteinase-9 (MMP-9) mRNA and protein expression were evaluated by real-time PCR and Western blot.The NF-kB activity was determined by dual-luciferase reporter assay system.The invasion ability of pancreatic cancer cells was evaluated by transwell invasion chamberassay.Results The TLR4 mRNA expressions in parental cells group,TP group,LPS group and TP + LPS group were 0.41 ± 0.06,0.46 ± 0.10,0.20 ± 0.04,0.25 ± 0.06 ; the TLR4 protein expressions were 0.55 ±0.06,0.55 ±0.06,0.18 ±0.04,0.13 ±0.00; the activities of NF-kB were 13.0 ±3.0,31.6 ±4.3,7.3 ±1.5 and 10.8 ± 2.1,and the numbers of invasion cell were (56.8 ± 8.6),(104.5 ± 12.8),(32.0 ± 5.7) and (46.8 ± 7.0) ; the MMP-9 mRNA expressions were 0.36 ± 0.05,0.58 ± 0.07,0.18 ± 0.03,0.30 ± 0.004 ;the MMP-9 protein expressions were 0.31 ± 0.04,0.53 ± 0.08,0.11 ± 0.02,0.15 ± 0.00.In LPS group,TLR4 mRNA and protein expressions were not statistic significant when compared with those in parental cells group,but the activities of NF-kB,the numbers of invasion cell,MMP 9 mRNA and protein expressions were statistically increased when compared with those in parental cells group (t =8.654,7.593,6.655,4.982,P ＜0.01).TLR4 mRNA and protein expressions,activities of NF-kB,the numbers of invasion cell,MMP 9 mRNA and protein expressions in TP group were significantly lower than those in parental cells group (t =-7.609,-9.948,-4.176,-5.915,-8.179,-9.948,P＜ 0.01).TLR4 mRNA and protein expressions,activities of NF-kB,the numbers of invasion cell,MMP 9 mRNA and protein expressions in TP +LPS group were significantly lower than those in LPS group (t =-4.437,-14.805,-10.506,-9.700,-9.055,-8.932,P＜ 0.01).Conclusions TP can inhibit pancreatic cancer cell invasion,and the mechanism is related to the inhibition of TLR4/NF-kB signaling pathway and down-regulation of MMP-9 expression.

Evidence has indicated that low doses of anti-tumor regimens can induce cell apoptosis in vitro, although different regimens induce apoptosis by different mechanism and pathway. In recent years, new tumor treatment strategy has been mainly focused on inducing tumor cell apoptosis. The present review discusses the advantages and disadvantages of inducing tumor cell apoptosis. The benefit of inducing apoptosis is not to cause inflammatory reaction, but as its disadvantage, it inhibits immune responses, and the phagocytosis of apopotic bodies may result in horizontal transfer of genes (including oncogenes and other oncogenic materials), which can be one of the causes of tumor relapse. This paper proposes that the tumor treatment strategy should be turn into promoting tumor cell necrosis and inducing anti-tumor immune responses.
Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Humans ; Necrosis ; chemically induced ; Neoplasms ; drug therapy ; immunology ; pathology

Objective To observe the relationship between Toll-like receptor 4 (TLR4) and the sensitivity of PANC1 cells to gemcitabine (GEM),and to analyze the potential mechanism.Methods PANC1 cells were divided into GEM group,lipopolysaccharide (LPS) + GEM group and TLR4-siRNA + GEM group.GEM group was treated by GEM alone.LPS + GEM group was pretreated with 1 mg/L LPS for 4 h and then treated by GEM.TLR4-siRNA + GEM group was transfected with 100 pmol/mL TLR4-siRNA for 4 h and then treated by GEM.The untreated cells were used as the control group.MTT method was used to detect the cell proliferation.Morphological changes and apoptosis rate of the cells were examined by Hoechst33258 staining and flow cytometry,respectively.The protein expression of TLR4,phosphorylated AKT (p-AKT) and activated Caspase-3 were detected by Western blot.Results The median inhibition concentration (ICs0) of GEM in the GEM group,LPS + GEM group and TLR4-siRNA + GEM group was (8.9 ± 0.32),(14.21 ±0.95),(3.96 ± 0.27) mg/L,respectively.The IC50 in LPS + GEM group was significantly higher than that in GEM group (P ＜ 0.01),and the IC50 of GEM in TLR4-siRNA + GEM group was significantly lower than that in GEM group (P ＜0.01).Compared with that in GEM group,the cells with typical apoptotic morphological changes were decreased in LPS + GEM group,which was increased in TLR4-siRNA + GEM group.The apoptotic rate in control group,GEM group,LPS + GEM group,TLR4-siRNA + GEM group was (2.1 ± 0.3) ％,(15.1 ± 2.3) ％,(9.8 ± 1.5) ％,(22.9 ± 3.1) ％,respectively.Compared with that in GEM group,the cells apoptotic rate was significantly reduced in LPS + GEM group (P ＜0.01),which was significantly increased in TLR4-siRNA + GEM group (P ＜0.01).TLR4 protein level in the 4 groups was 0.83 ±0.08,0.81 ±0.07,0.85 ±0.07 and 0.16 ±0.03;p-AKT protein level 0.61 ±0.05,0.36 ±0.03,0.73 ± 0.07 and 0.21 ± 0.02;activated Caspase-3 protein level was 0.66 ± 0.05,0.73 ± 0.07,0.45 ± 0.04 and 0.91 ± 0.07,respectively.The expression of TLR4 and p-AKT in TLR4-siRNA + GEM group was significantly lower than that in GEM group (P ＜0.01),while the expression of activated Caspase-3 protein was increased significantly (P ＜ 0.05).Compared with the GEM group,the expression of p-AKT protein in LPS + GEM group was significantly increased (P＜0.01),and the expression of activated Caspase-3 protein was significantly decreased (P＜0.01).Conclusions TLR4 can inhibit the sensitivity of pancreatic cancer PNAC1 cells to GEM,and the mechanism is related to the activation of PI3K/AKT pathway and downregulation of activated Caspase-3.

Female ; Humans ; Infant ; Mitochondrial Proton-Translocating ATPases ; deficiency ; genetics ; Mutation

Objective To provide a good anatomical foundation for transposition of the branch of median nerve superficial flexor muscle repairing the movement branch of ulnar nerve to recover the function of intrinsic muscle, by anatomic study of the muscle branch of the superficial flexor muscle of the median nerve and the movement branch of ulnar nerve. Methods Twenty adult upper limb specimens immersed fixed by formalin were elected and expose the midian nerve and ulnar nerve. Then every anatomical index was measured. Simulate to manipulate that the branch of superficial flexor muscle repair the motor b ranch of ulnar nerve. Calculate the number of myelinated nerve fibers of the branch of superficial flexor muscle. Results The distance between the position into muscle and styloid process of radius and styloid process of ulna: (21.4±1.8)mm, the distance that can be separated: (27.1±1.2)mm, the transverse diameter: (1.2±0.2)mm, anteroposterior diameter: (0.7 ± 0.1 )mm. No injury separated distance between the sensory branch and motor branch of ulnar nerve: (7.1 ± 0.7)cm. The 4th muscular branches of median nerve flexor digitorum superficialis was 1378.9± 107.9. Conclusion The 4th muscular branches of median nerve flexor digitorum superficialis can be used to repair the motor branch of the ulnar nerve to recover the function of intrinsic muscle of hand.

Relapse, which puzzled several generations of hematologists, is the bottle-neck of radical treatment for leukemias. The progress of Human Microbiome Project at the beginning of 21st century suggested that human body was a super-organism constituted by the core of human cells and symbiotic microorganisms. The elucidation and characterization of endogenous retrovirus and prion protein suggested the possible effects of co-evolutional microorganisms on human health. Recently, the elucidation of the roles of tunneling nanotubes in intercellular communication and transportation suggested a novel way for cellular communication and transport of oncogenic materials. The role and significance of in vivo cell fusion have been studied in more detail. On the other hand, donor cell leukemia was reported. All of these approaches provide novel insights for studying the mechanism of leukemia relapse. Based on previous work, the authors suggest the hypothesis: there are two possible mechanisms for the relapse of leukemias: the minimal residual disease (MRD) and intercellular transportation of oncogenic materials.
Cell Fusion ; Humans ; Leukemia ; pathology ; Neoplasm, Residual ; pathology ; Recurrence

Objective To compare the mid-term clinical results of primary implantation of a selfexpanding nitinol stent and primary percutaneous transluminal angioplasty (PTA) for long range arteriosclerosis obliterans of the superficial femoral artery. Methods From December 2005 to February 2007,109 patients who had moderate-severe claudication or chronic limb ischemia (CLI)due to stenosis or occlusion of the superficial femoral artery were treated by endovascular technology. 53 patients (73 limbs) were treated by primary stenting and 56 patients (76 limbs)by PTA. We compared the clinical data of stenting group and PTA group at 6,12 and 24 months. Results The mean length of the treated segment was (16± 8)cm in the stenting group and(15±7)cm in the PTA group. At 6 months, the rate of restenosis on duplex ultrasonography was 13.7% in stenting group and 30.2% in PTA group (X2=4.09, P<0.05);at 12 months the restenosis rates were 25.5% in stenting group and 46.9% in PTA group(X2=4.75, P<0.05);at 24 months the restenosis rates on duplex ultrasonography were 38.1% in stenting group and 65.9% in PTA group(X2=6.66, P<0.01). Rutherford stages in stenting group were significantly better than those in PTA group. Conclusion In the mid-term, primary implantation of a self-expanding nitinol stent yielded results that were superior to those with PTA for arteriosclerosis obliterans or long range stenosis of the superficial femoral artery.

Objective To analyze the mid and long term results of endovascular procedures for diabetic below-the-knee lesions.Methods We retrospectively analyzed data of 762 patients (797 limbs in Wagner Grade 1 to 4) who suffered ischemic symptoms of diabetic below-the-knee lesions,admitted for endovascular procedures from Aug 2005 to May 2012.Technical success rate and complications,anklebrachial index,wound healing,limb salvage and survival were evaluated.Results 762 patients (797 limbs) underwent 857 endovascular procedures with immediate technical success in 805 times (success rate 93.93％).30 day complications included 33 cases (4.33％) of hematoma or pseudoaneurysm related to access site,20 cases (2.62％) of vasovagal reflex,38 cases (4.99％) of belowthe-knee arteriorrhexis,75 cases (9.84％) of inflow-limited dissection,23 cases (3.02％) of arterial embolism,4 cases (0.52％) of blue-toe sign.Postoperative mean ABI was improved significantly,from 0.33 ± 0.20 to 0.68 ± 0.17,P ＜ 0.005.722 patients (751 limbs) were followed up for more than 12 months (35 ± 23 months),wound healing rate within 3 months was 72.28％,limb salvage rate after 12 months was 85.62％,survival rate was 95.01％.Conclusions Immediate technical success rate of endovascular procedures for diabetic below-the-knee lesions is high,complications rate is low,medium and long term efficacy is satisfactory.

Objective To observe the primary clinical results of Gore （R） Viabahn endoprosthesis for atherosclerotic stenosis or occlusion in superficial femoral artery (SFA).Methods From March 2013 to November 2013,45 consecutive patients (54 limbs) who had ischemic symptom due to stenosis or occlusion of superficial femoral artery were treated by endovascular deployment of Gore （R） Viabahn endoprosthesis.We observed patency rate,improvement of Rutherford classification and ankle-brachial index,limb salvage rate and survival rate after 1,3 and 6 months.Results 43 patients (52 limbs)were followed-up,among which 32 patients(38 limbs)belonged to TASC Ⅱ type C or type D lesion.The mean (± SD) length of treated segment in 52 limbs was(19 ±4)cm.The rate of postoperative patency on duplex ultrasonography:one month,3 months and 6 months were 96.15％ (50 limbs),92.31％ (48 limbs)and 90.38％ (47 limbs) ; Ankle-brachial index increased from(0.32 ±0.20)to(0.68 ±0.18) (t =3.180,P ＜ 0.005) after 6 months ; at 6 months limb salvage rate was 96.08 percent (49 limbs),and survival rate was 97.67％ (42 patients).Conclusions Implantation of Gore （R） Viabahn endoprosthesis in atherosclerotic stenotic or occluded superficial femoral artery safely achieves a definite primary clinical effectiveness.

Objective To estimate the security and effectiveness of double access technique to treat chronic total occlusion in lower extremity artery. Method Fifty four patients,who had lower limb arteriosclerosis and accepted failed endovascular treatment because of unable to reenter true lumen through antegrade access,were treated immediately in double access style to recanalize occluded artery (including 27 patients of dorsalis pedis artery puncture,17 patients of posterior tibial artery punctrue,5 patients of proximal anterior tibial artery puncture and 5 patients of distal superficial femoral artery puncture). Evaluate the revascularization rate,complication rate and 6-month limb salvage rate of double access technique,t test was used to compare the ABI.Results Revascularization rate was 98.2％ (53/54),average puncture frequency of every case was 6 ( the first access was 2 in every patient,the second access was 4 in every patient ),average treatment time was 167 minutes,complication rate was 5.6％ (3/54),2 patients were occlusion in puncture segment of the second access,1 patient was hematoma in the second access point. 6-month ABI after intervention increased from 0.44 ±0.13 to 0.69 ±0.15,6-month limb salvage rate was 100％. Conclusion For chronic total occlusion,double access is a feasible technique to deal with unable to reenter true lumen throughantegrade access,the complication rate is low,the primary limb salvage rate is relatively high.