Loss of GRB2 associated binding protein 1 in arteriosclerosis obliterans promotes host autophagy.

BACKGROUND: Arteriosclerosis obliterans (ASO) is a major cause of adult limb loss worldwide. Autophagy of vascular endothelial cell (VEC) contributes to the ASO progression. However, the molecular mechanism that controls VEC autophagy remains unclear. In this study, we aimed to explore the role of the GRB2 associated binding protein 1 (GAB1) in regulating VEC autophagy. METHODS: In vivo and in vitro studies were applied to determine the loss of adapt protein GAB1 in association with ASO progression. Histological GAB1 expression was measured in sclerotic vascular intima and normal vascular intima. Gain- and loss-of-function of GAB1 were applied in VEC to determine the effect and potential downstream signaling of GAB1. RESULTS: The autophagy repressor p62 was significantly downregulated in ASO intima as compared to that in healthy donor (0.80 vs. 0.20, t = 6.43, P < 0.05). The expression level of GAB1 mRNA (1.00 vs. 0.24, t = 7.41, P < 0.05) and protein (0.72 vs. 0.21, t = 5.97, P < 0.05) was significantly decreased in ASO group as compared with the control group. Loss of GAB1 led to a remarkable decrease in LC3II (1.19 vs. 0.68, t = 5.99, P < 0.05), whereas overexpression of GAB1 significantly led to a decrease in LC3II level (0.41 vs. 0.93, t = 7.12, P < 0.05). Phosphorylation levels of JNK and p38 were significantly associated with gain- and loss-of-function of GAB1 protein. CONCLUSION: Loss of GAB1 promotes VEC autophagy which is associated with ASO. GAB1 and its downstream signaling might be potential therapeutic targets for ASO treatment.

Upregulation of CBP by PLY can cause permeability of blood-brain barrier to increase meningitis.

BACKGROUND: Streptococcus pneumoniae causes many human diseases including bacterial meningitis. Previous study proposed that pneumolysin (PLY), a cytotoxin from pneumococcus, is related to the infection across blood-brain barrier (BBB). However, the mechanism of how PLY break through BBB remains elusive. The present study showed that PLY can increase the permeability of BBB both in vitro and in vivo in our experiments. RESULTS: Further we found out that PLY leads to the high expression of CERB-binding protein (CBP) which can lead to releasing of tumor necrosis factor α then enhance apoptosis of cells which is a significant factor leading to permeabilization of BBB. CONCLUSION: Our findings demonstrate that CBP plays an important role in the pneumococcus infection in the brain and could be a potential therapeutic target against pneumococcal meningitis.

Open surgical treatment for rare epithelioid hemangioendothelioma of the common femoral vein.

Epithelioid hemangioendothelioma is a very rare disease affecting big blood vessels, especially veins. There have been very few articles describing the disease. We hereby present the case of a 56-year-old woman presenting with lower limb edema, who was initially being treated for residual thrombus in the common femoral vein but was eventually diagnosed with epithelioid hemangioendothelioma (EHE). The common femoral vein was resected and reconstructed using the external jugular vein. No additional therapy was administered. In this article, previous literature about EHE has been reviewed and oncologic principles have been discussed.

Corydalis bungeana Turcz. attenuates LPS-induced inflammatory responses via the suppression of NF-κB signaling pathway in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Corydalis bungeana Turcz. (C. bungeana) is one of traditionally used medicines in China and possesses various biological effects, such as anti-inflammatory, antibacterial activity and inhibition of the immune function of the host. AIM OF THE STUDY: we studied the anti-inflammatory effect and molecular mechanism involved of C. bungeana both in vitro and in vivo model system in which the inflammatory response was induced by LPS treatment. MATERIALS AND METHODS: Anti-inflammatory activity of C. bungeana was investigated by LPS-induced RAW 264.7 macrophages and BALB/c mice. The production and expression of pro-inflammatory cytokines were evaluated by Griess reagent, ELISA kits and RT-qPCR, respectively. Phosphorylation status of IκBα and p65 was illustrated by western blot assay. RESULTS: C. bungeana reduced the secretion of NO, TNF-α, IL-6 and IL-1ß through inhibiting the protein expression of iNOS, TNF-α, IL-6 and IL-1ß in vitro and in vivo. Western blot analysis suggested that C. bungeana supressed NF-κB activation via regulating the phosphorylation of IκBα and p65. Immunohistochemical assay also demostrated the histological inflammatory change in liver tissue. CONCLUSIONS: The results indicate that C. bungeana supresses the activation of NF-κB signaling pathway through inhibiting phosphorylation of IκBα and p65, which results in good anti-inflammatory effect. In addition, C. bungeana attenuates inflammatory reaction by supressing the expression of various inflammatory cytokines both in vivo and in vitro.

Nauclea officinalis inhibits inflammation in LPS-mediated RAW 264.7 macrophages by suppressing the NF-κB signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Nauclea officinalis has been traditionally used in China for the treatment of fever, pneumonia and enteritidis etc. This study aims to investigate effects of N. officinalis on the inflammatory response as well as the possible molecular mechanism in LPS-stimulated RAW 264.7 murine macrophage cells. MATERIALS AND METHODS: Anti-inflammatory activity of N. officinalis (10, 20, 50, and 100µg/mL) was investigated by using LPS-induced RAW 264.7 macrophages. The NO production was determined by assaying nitrite in culture supernatants with the Griess reagent. The levels of TNF-α, IL-6 and IL-1ß in culture media were measured with ELISA kits. Real time fluorescence quantitative PCR was detected for mRNA expression of iNOS, TNF-α, IL-6 and IL-1ß. Western blot assay was performed to illustrate the inhibitory effects of N. officinalis on phosphorylation of IκB-α and NF-κB p65. RESULTS: Treatment with N. officinalis (10-100µg/mL) dose-dependently inhibited the production as well as mRNA expression of NO, TNF-α, IL-6 and IL-1ß in RAW 264.7 macrophages. Western blot assay suggested that the mechanism of the anti-inflammatory effect was associated with the inhibition of phosphorylation of IκB-α and NF-κB p65. CONCLUSIONS: The results indicated that N. officinalis potentially inhibited the activation of upstream mediator NF-κB signaling pathway via suppressing phosphorylation of IκB-α and NF-κB p65 to inhibit LPS-stimulated inflammation.

[Molecular mechanisms of antithrombin deficiency caused by novel double heterozygous mutations].

OBJECTIVE: To explore the molecular mechanisms of antithrombin (AT) deficiency caused by novel double heterozygous mutations. METHODS: Wild-type and mutant AT cDNA expression plasmids (ATwt, AT-c.134G > A, AT-c.342T > G, AT-c.134G > A and AT-c.342T > G) were transfected into HEK293T cells. Western blot was used to detect the AT:Ag in cell lysates. Homology was used to reestablish 3-D spatial structure of AT. Laser confocal assay was utilized to analyze the distribution of AT in endoplasmic reticulum (ER). RESULTS: Compared to the wild-types, the AT expression of HEK293T cells sharply increased when they were transfected by AT-c.342T > G or AT-c.134G > A and c.342T > G. Homology modeling showed that the mutation (AT-c.342T > G) caused a decreased distance between Arg and surrounding bases as Arg's side chain was significantly longer than Ser's. The mutation of 13th base pair decreases the distance between AT and heparin. Laser confocal assay showed that the AT protein concreted in HEK293T cells when they were transfected by mutant plasmids (AT-c.134G > A and c.342T > G) and aggregated in ER. CONCLUSIONS: The main molecular mechanism of AT deficiency in this pedigree is the disturbed AT secretion as a result of the mutation of AT-c.342T > G.

Endovenous laser ablation of great saphenous vein with ultrasound-guided perivenous tumescence: early and midterm results.

BACKGROUND: Endovenous laser ablation (EVLA) is an improved method to treat varicose great saphenous veins (GSV) with a high satisfactory rate. This study aimed to evaluate the efficiency and safety of treatment by EVLA procedures with ultrasound-guided perivenous tumescence. METHODS: Thirty-one patients (31 limbs) with symptomatic varicose vein primary to chronic venous insufficiency (CVI) treated with EVLA were prospectively studied. The entire procedure was performed under ultrasound-guided tumescent local anesthesia. The patients were evaluated with a 18 month follow-up postoperation using clinical examination and venous duplex ultrasonography. Pain scores and quality of life (QOL) were recorded using visual analog scale (VAS) and the chronic venous insufficiency questionnaire (CIVIQ) at 1 week, 1 month, and 12 months after operation. RESULTS: All patients tolerated EVLA procedure well. The overall success occlusion rates of GSV were 92%, 94%, and 94% at 1, 12, and 18 months follow-up, respectively. The score of CIVIQ one week preoperation was 69.14 ± 11.44 while that of CIVIQ one month postoperation was 85.32 ± 4.89. The life quality has significantly improved after the operation of EVLA (t = 12.71, P < 0.05). The VAS one month after treatment was lower than 1 week before therapy (t = 8.048, P < 0.05). Major complications such as deep vein thrombosis and skin burns were not found. Most of the complications were minor and improved quickly. CONCLUSIONS: This refinement type of EVLA procedure is a safe and effective treatment with a high satisfaction rate; it displayed noteworthy features including shortening hospitalization, early ambulant activity, and preferable occlusion rates.

[The efficacy analysis of endovascular intervention therapy for transatlantic inter society consensus type C and type D femoropopliteal arteriosclerosis lesion].

OBJECTIVE: To evaluate the efficacy of the endovascular intervention for transatlantic inter society consensus (TASC) type C and type D femoropopliteal artery disease. METHODS: We conducted a retrospective analysis on 95 cases (98 lower limbs) with TASC type C and type D femoropopliteal artery arteriosclerosis lesion treated by percutaneous transluminal angioplasty and/or primary stent implantation from January 2007 to April 2009. We used ankle brachial index (ABI), Fontaine stages, limb salvage percentage and primary patency to evaluate the efficacy of the endovascular intervention therapy. RESULTS: The technical success rate of the 98 limbs was 94.9%, the perioperation mortality was 4.2% and the total amputation rate was 5.1%. 81 cases (84 limbs) were followed-up for a mean time of (13 ± 7) months, whose average ABI in dorsalis pedis artery and posterior tibial artery were 0.58 ± 0.22 and 0.60 ± 0.21 and increased 0.14 ± 0.25 and 0.13 ± 0.22 respectively than the ABI before intervention therapy. The statistical analysis showed a significant difference in ABI. The limbs of critical limb ischemia (CLI) were of 16.4% in the follow-up period and of 73.5% before the intervention therapy. The statistical results showed a significant difference in the percentage of CLI. CONCLUSION: Percutaneous endovascular intervention is an effective and minimally invasive method, and has a curative clinical efficacy to treat TASC type C and type D femoropopliteal artery disease.

Flow-injection biamperometric direct determination of calcium dobesilate in irreversible couple system.

A flow-injection biamperometric method for direct determination of calcium dobesilate had been proposed based on biamperometric detection for irreversible couple. The detection was realized by coupling the oxidation of dobesilate at one platinum wire electrode with the reduction of MnO(4)(-) at another one with the applied potential difference of 0 V between two platinum wire electrodes. Dobesilate was determined in the range of 4.0 x 10(-6) to 1.0 x 10(-4) M with the detection limit of 8.0 x 10(-7) M (S/N=3). The relative standard derivation of 1.7% was obtained for 24 successive determinations of 4.0 x 10(-5) M dobesilate. The proposed method had been shown to be sensitive, simple and rapid.