Anti-Nasopharyngeal carcinoma mechanism of sanguinarine based on network pharmacology and molecular docking.

BACKGROUND: The purpose of this study was to investigate the mechanism of sanguinarine (SAN) against nasopharyngeal carcinoma (NPC) by means of network pharmacology, molecular docking technique, and experimental verification. METHODS: The SAN action targets were predicted using the Swiss Target Prediction database, the related NPC targets were determined using the GEO database, and the intersection of drug and disease pathway targets were considered to be the potential targets of SAN against NPC. The target-protein interaction network map was constructed using the STRING database, and the core target genes of SAN against NPC were obtained via topological network analysis. "R" language gene ontology (GO) function and Kyoto encyclopedia of genes and genome (KEGG) pathway enrichment analyses were used to dock the core target genes with SAN with the help of AutodockVina. Cell proliferation was detected using MTT and xCELLigence real-time cell analysis. Apoptosis was identified via Hoechst 33342 staining, JC-1 mitochondrial membrane staining, and annexin V-FITC/PI double fluorescence staining, while protein expression was quantified using western blotting. RESULTS: A total of 95 SAN against NPC targets were obtained using target intersection, and 8 core targets were obtained by topological analysis and included EGFR, TP53, F2, FN1, PLAU, MMP9, SERPINE1, and CDK1. Gene ontology enrichment analysis identified 530 items, and 42 items were obtained by Kyoto encyclopedia of genes and genome pathway enrichment analysis and were mainly related to the PI3K/AKT, MAPK, and p53 signaling pathways. Molecular docking results showed that SAN had good binding activity to the core target. SAN inhibited the proliferation of NPC cells, induced apoptosis, reduced the expression levels of survivin and Bcl2, and increased the expression levels of Bax and cleaved caspase-8. It also decreased the expression levels of the key proteins p-c-Raf, p-MEK, and p-ERK1/2 in the MAPK/ERK signaling pathway in NPC cells. CONCLUSION: SAN inhibits the proliferation and induces the apoptosis of NPC cells through the MAPK/ERK signaling pathway.

Prevention of in-stent restenosis with endothelial progenitor cell (EPC) capture stent placement combined with regional EPC transplantation: An atherosclerotic rabbit model.

BACKGROUND: Even with drug-eluting stents, the risk of in-stent restenosis (ISR) remains high. The goal of this study was to investigate the use of an endothelial progenitor cell (EPC) capture stent plus regional EPC transplantation to reduce the ISR rate. METHODS: Endothelial progenitor cell capture stents were fabricated using fibrin gel and anti-CD34 plus anti-VEGFR-2 dual antibodies. Twenty male New Zealand white rabbits established as an atherosclerotic model were randomly divided into two groups: group 1 (n = 10), in which EPC capture stents were deployed into the right iliac artery; and group 2 (n = 10), in which sirolimus-eluting stents were placed. In both groups, EPCs were transplanted into target vessels beyond the stents, with outflow blocked. Radiologic-pathologic correlation outcomes were reviewed after 2 months. RESULTS: The technical success rate of EPC capture stent placement plus EPC transplantation was 100%. The ISR rate in group 1 was lower than in group 2 (1/10 vs. 4/10; p > 0.05). Minimal luminal diameters were larger in group 1 than in group 2 (computed tomographic angiography, 1.85 ± 0.15 mm vs. 1.50 ± 0.20 mm; duplex ultrasound, 1.90 ± 0.10 mm vs. 1.70 ± 0.30 mm; p > 0.05). Transplanted EPCs were tracked positively only in group 1. Pathologic analysis demonstrated neointimal hyperplasia thickness of 0.21 ± 0.09 mm in group 1 vs. 0.11 ± 0.07 mm in group 2 (p < 0.05). CONCLUSION: Endothelial progenitor cell capture stent placement plus local EPC transplant decreases the ISR rate through thrombosis reduction rather than through neointimal hyperplasia inhibition.

Evaluation of Visceral Adipose Tissue Oxygenation by Blood Oxygen Level-Dependent MRI in Zucker Diabetic Fatty Rats.

OBJECTIVE: This study aimed to investigate the feasibility of blood oxygen level-dependent magnetic resonance imaging (BOLD-MRI) to evaluate visceral adipose tissue (VAT) oxygenation in Zucker diabetic fatty (ZDF) rats and its associations with systemic metaflammation. METHODS: Five-week-old ZDF rats and Zucker lean (ZL) rats were fed a high-fat diet (HFD) for 18 weeks. A baseline BOLD-MRI scan of perirenal adipose tissue was performed after 8 weeks of HFD feeding, and then the rats were randomized to receive pioglitazone or a vehicle for the following 10 weeks. At sacrifice, BOLD-MRI scan, Hypoxyprobe-1 injection, and circulating T helper 17 (Th17), regulatory T (Treg) cells, and monocyte subtype flow cytometry analysis were performed. RESULTS: HFD feeding led to a significant increase in VAT BOLD-MRI R2* signals (20.14 ± 0.23 per second vs. 21.53 ± 0.20 per second; P = 0.012), an indicator for decreased oxygenation. R2* signal was significantly correlated with VAT pimonidazole adduct-positive area, insulin resistance, Th17 and Treg cells, CD43 + and CD43+ + monocyte subtypes, and VAT macrophage infiltration. Pioglitazone treatment improved the insulin resistance and was associated with a delayed progression of VAT oxygenation. CONCLUSIONS: This work demonstrated the feasibility of BOLD-MRI for detecting the VAT oxygenation status in ZDF rats, and the BOLD-MRI signals were associated with insulin resistance and systemic metaflammation in ZDF rats during the development of obesity.

Percutaneous mechanical thrombectomy for acute massive lower extremity deep venous thrombosis.

PURPOSE: To evaluate the clinical safety and effectiveness of percutaneous mechanical thrombectomy in patients with acute massive lower extremity deep venous thrombosis. MATERIALS AND METHODS: Twenty-five consecutive patients with acute massive lower extremity deep venous thrombosis were included in this retrospective study. An inferior vena cava filter was placed prophylactically to protect against pulmonary embolism in each patient. Percutaneous mechanical thrombectomy was performed using a 7F Amplatz thrombectomy device in an angiography suite through ipsilateral popliteal vein access. Anticoagulant therapy lasted for at least 6 months. Follow-up data included 1 year's color duplex sonography and clinical interviews. RESULTS: Successful placement of an inferior vena cava filter was achieved in all 25 (100%) patients. Twenty-two patients (88%) were clinically asymptomatic within 24 hours, whereas the remaining 3 patients (12%) showed moderate improvement within 48 hours. Venogram at discharge showed grade III lysis in 23 patients (92%) and grade II lysis in 2 patients (8%). No serious complications were reported during hospitalization in this study. At 1-year follow-up, no recurrent deep venous thrombosis was reported; 1 patient developed a mild postthrombotic syndrome. CONCLUSION: Percutaneous mechanical thrombectomy is a safe and effective treatment for acute massive lower extremity deep venous thrombosis and shows promising clinical mid-term results.

Seeding endothelial progenitor cells on a self-expanding metal stent: an in vitro study.

PURPOSE: To demonstrate the feasibility of seeding a self-expanding metal stent with endothelial progenitor cells to enhance rapid reendothelialization, which is postulated to prevent local thrombus formation and restenosis after vascular intervention. MATERIALS AND METHODS: Endothelial progenitor cells and fibrinogen were isolated from the peripheral blood of a domestic swine and then cultured and identified. Ten self-expanding nitinol stents were incubated in the culture medium with a cell concentration of 1 x 10(6)/mL with (n = 5, study group) or without (n = 5, control group) fibrin gel (5 mg/mL fibrinogen and 0.10 NIHU/mL thrombin) for 24 hours. The cell coverage of the stents was documented with en face photography and scanning electron microscopy. After simulated use in vitro, the cells were removed from each stent, counted with a cytometer, sequentially cultured for three passages, and identified again to compare their properties with those of the original seeding line. RESULTS: After seeding the stent with the combination of endothelial progenitor cells and the fibrin gel coating, the stents took on a tube-like appearance with a confluent monolayer membrane. After digestion with trypsin, a mean of 2.5 x 10(5) +/- 1.3 cells were obtained from the fibrin gel stent (study group); fewer cells (4 x 10(4) +/- 1.5) were recovered from the bare stents (control group) (P < .01). The recovered cells, after amplification with culture, demonstrated the properties of the original endothelial progenitor cells. CONCLUSIONS: An endothelial progenitor cell-coated stent can be successfully fabricated by using fibrin gel as the bonding agent in vitro. Further in vivo research is warranted.

Transjugular intrahepatic portosystemic shunt with an autologous endothelial progenitor cell seeded stent: a porcine model.

RATIONALE AND OBJECTIVES: To evaluate the efficacy of a self-expanding metal stent seeded with autologous endothelial progenitor cells (EPCs) for preventing in-stent stenoses in transjugular intrahepatic portosystemic shunt (TIPS) in a swine model. MATERIALS AND METHODS: TIPS was performed in 18 young adult pigs, using a self-expanding nitinol stent (control, n = 8) and an autologous EPC-seeded stent (treatment, n = 10). All pigs were sacrificed at 2 weeks post-TIPS procedure. Portography was performed immediately before the euthanasia. Gross, microscopic, and immunohistochemistry of the TIPS tract specimens were examined. The proliferative response of the shunt was quantified histologically. RESULTS: TIPS was performed successfully in 16 swine, 2 animals died during the procedure. Another pig died of unknown causes 2 days post-procedure. At day 14 follow-up, portography and necropsy of the 15 remaining swine demonstrated that five shunts occluded and one shunt was stenotic (80%) in the control group (n = 6). Five shunts remained patent, two shunts were stenosed (50%, 70%), and the remaining two shunts were occluded in the treatment group (n = 9). The patency rate was significantly lower in the control group than in the treatment group, 0% versus 55.6% (P = .03). Histological analyses showed a significantly greater pseudointimal hyperplasia in the TIPS track of the control group than that of the treatment group (P < .05). Intact endothelium was documented in the lumina of all the EPC-implanted stent group. CONCLUSIONS: The EPC-seeded metal stent is feasibly fabricated in vitro and improves the patency in TIPS in a porcine model.

In vivo tracking of dual-labeled mesenchymal stem cells homing into the injured common carotid artery.

The aim of this study is to conduct in vivo, noninvasive magnetic resonance imaging of labeled rat bone mesenchymal stem cells (BMSCs) as they home into the site of injured common carotid artery following allograft transplantation. Our study was approved by the Institutional Committee on Animal Research. Purified rat BMSCs were dual labeled with superparamagnetic iron oxide (SPIO) particle and fluorescent DiI dye, and subsequently transplanted into recipient rats injured in the left common carotid arteries. Immediately before and 3 hr, 3, 7 and 12 days after transplantation, the labeled cells were monitored in vivo using a 7T micromagnetic resonance imaging (7T micro-MRI) scanner. The signal-to-noise ratios (SNRs) at the injured sites were corroborated with histological examination using Prussian blue staining and fluorescent imaging. Rat BMSCs were labeled with SPIO and DiI at 100% efficiency. When compared with the baseline level before transplantation, the SNR decreased significantly on Days 3 and 7 after injection in the experimental group (Dunnet t test, P < 0.05), whereas insignificant differences were observed after 3 hr and 12 days (Dunnet t test, P > 0.05). In the control group, no significant differences in SNR were found among different time points (ANOVA, P > 0.05). Histological analyses illustrated that red fluorescence and Prussian blue-positive cells were mainly distributed around the lesion areas of injured common carotid arteries. Rat BMSCs can be efficiently labeled with SPIO and DiI, and the directional homing of labeled cells to the site of injured common carotid arteries after intravascular transplantation could be tracked in vivo with 7T micro-MRI.

Percutaneous mechanical thrombectomy combined with catheter-directed thrombolysis in the treatment of symptomatic lower extremity deep venous thrombosis.

PURPOSE: To evaluate the efficacy of percutaneous mechanical thrombectomy (PMT) combined with catheter-directed thrombolysis (CDT) in the treatment of massive symptomatic lower limb deep venous thrombosis (DVT). MATERIALS AND METHODS: One hundred and three clinically confirmed DVT patients were discharged from our institution. Sixteen patients with massive lower limb DVT were included in this retrospective study. After prophylactic placement of inferior vena cava filters (IVCFs), percutaneous mechanical thrombectomy (ATD, n=10; Straub, n=6) and catheter-directed thrombolysis were performed in all patients. Complementary therapy included percutaneous transluminal venous angioplasty (PTA, n=3) and stent placement (n=1). The doses of thrombolytic agents, length of hospital stay, peri-procedure complications and discharge status were reviewed. Oral anticoagulation was continued for at least 6 months during follow-up. RESULTS: The average hospital stay was 7 days. The technical success rate (complete and partial lysis of clot) was 89%, the other 11% patients only achieved less than 50% clot lysis. The mean dose of urokinase was 3.3 million IU. There were no significant differences of clinical outcome between the ATD and Straub catheter group. The only major complication was an elderly male who experienced a fatal intracranial hemorrhage while still in the hospital (0.97%, 1/103). Minor complications consisted of three instances of subcutaneous bleeding. No transfusions were required. Vascular patency was achieved in 12 limbs during follow-up. No pulmonary emboli occurred. There is one recurrent DVT 4.5 months after the treatment. CONCLUSIONS: Percutaneous mechanical thrombectomy combined with catheter-directed thrombolysis is an effective and safe method for the treatment of symptomatic DVT. A randomized prospective study is warranted.

[In vitro MR imaging of Fe2O3-arginine labeled heNOS gene modified endothelial progenitor cells].

OBJECTIVE: To explore the feasibility of in vitro magnetic resonance imaging on Fe2O3-arginine labeled heNOS gene modified endothelial progenitor cells (EPCs). METHODS: Fe2O3 was incubated with arginine to form Fe2O3-arginine complex. Rabbit peripheral blood mononuclear cells (MNCs) were isolated and EPCs were isolated by adherence method, expanded and modified with heNOS gene using Lipofectamine 2000. After 48 hours, genetically modified EPCs were incubated with Fe2O3-arginine for 24 hours. Intracellular iron was detected by Prussian blue stain. The expression of heNOS gene was detected by Western blot. MTT assay was used to evaluate cell survival and proliferation of Fe2O3-arginine labeled heNOS-EPCs. Flow cytometry was used to measure cell apoptosis. The cells underwent in vitro MR imaging with various sequences. RESULTS: Iron-containing intracytoplasmatic vesicles could be clearly observed with Prussian blue staining, and the labeling rate of labeled heNOS-EPCs were similar to that of labeled EPCs (around 100%). Survival and apoptosis rates obtained by MTT and flow cytometry analysis were similar among labeled heNOS-EPCs, labeled EPCs and unlabeled EPCs with Fe2O3-arginine. The signal intensity on MRI was equally decreased in labeled heNOS-EPCs and labeled EPCs compared with that in unlabeled cells. The percentage change in signal intensity (DeltaSI) was most significant on T2*WI and DeltaSI was significantly lower in cells labeled for 7 days than that labeled for 1 days. CONCLUSIONS: The heNOS gene can be successfully transfected into rabbit peripheral blood EPCs using Lipofectamine2000. The heNOS-EPCs can be labeled with Fe2O3-arginine without significant change in viability and proliferation capacity. The labeled heNOS-EPCs can be imaged with standard 1.5 T MR equipment. The degree of MR signal intensity may indirectly reflect the cell count, growth and division status.