Rapid and facile detection of largemouth bass ranavirus with CRISPR/Cas13a.

Largemouth bass ranavirus (LMBV) is an epidemic disease that seriously jeopardizes the culture of largemouth bass（Micropterus salmoides）, and it has a very high incidence in largemouth bass. Once an outbreak occurs, it may directly lead to the failure of the culture, resulting in substantial economic losses, but there is no effective vaccine or special effective drug yet. Consequently, it is important to establish an accurate, sensitive, convenient and specific detection approach for preventing LMBV infection. The recombinant enzyme-assisted amplification (RAA) technology was used in combination with clustered regularly interspaced short palindromic repeats (CRISPR), and associated protein 13a (CRISPR/Cas13a) to detect LMBV. We designed RAA primers and CRISPR RNA (crRNA) that targeted the conserved region in the LMBV main capsid protein (MCP) gene, amplified sample nucleic acids using the RAA technology, performed CRISPR/Cas13a fluorescence detection and evaluated the sensitivity and specificity of the established method with qPCR as a control method. This technique was able to determine the results by collecting fluorescence signals, visualizing fluorescence by UV excitation and combining with lateral flow strips (LFS). The sensitivity and specificity of the established method were consistent with the qPCR method. Besides, it was performed at a constant temperature of 37 °C and the sensitivity of the reaction system was 3.1 × 101 copies/µL, with no cross-reactivity with other common aquatic pathogens. Further, the positive detection rate of the proposed method in 32 clinical samples was consistent with that of qPCR. In conclusion, our established RAA-CRISPR/Cas13 method for detecting LMBV is sensitive, simple and specific, which is applicable in the rapid on-site detection and epidemiological monitoring of LMBV.

Identification and application of an endophytic fungus Arcopilus aureus from Panax notoginseng against crop fungal disease.

Endophytic fungi are important microbial resources for developing novel antibacterial and antifungal drugs to prevent and control crop diseases. Panax notoginseng has been used as a Chinese medicinal herb for a long time, as it has various bioactivities. However, information on endophytic fungi isolated from Panax notoginseng is rare. In this study, an endophytic fungus known as SQGX-6, which was later identified as the golden hair fungus Arcopilus aureus, was isolated from Panax notoginseng. SQGX-6 was extracted using ethyl acetate, and the active components of the fungus were identified using ultra-performance liquid chromatography-mass spectrometry (UHPLC-MS). The antifungal and antioxidant activities of the extract were determined and evaluated in vitro and in vivo. SQGX-6 and its extract inhibited the growth of Corn stalk rot (Fusarium graminearum), Corn southern leaf blight (Helminthosporium maydis), and Tomato gray mold (Botrytis cinerea) in vitro. The free radical scavenging rates for 2,2-Diphenyl-1-pyridinyl hydrazide (DPPH) radical scavenging activity, 3-Ethylbenzothiazoline-6-Sulfonic Acid Radical scavenging (ABTS) activity were also downregulated by the SQGX-6 extract. In vivo, the SQGX-6 extract inhibited the mycelial growth rates of the three aforementioned fungi and downregulated malondialdehyde (MDA) content and upregulated peroxidase (POD) and phenylalanine ammonia-lyase (PAL) content in fruits, leading to significant reduction in damage to cherry tomatoes caused by Botrytis cinerea. UHPLC-MS was performed to identify various active substances, including Alkaloids, Azoles, Benzofurans, Coumarins, Flavonoids, Organic acids, Phenols, and plant growth regulators contained in the extract. These results suggested that the endophytic fungus SQGX-6 of Panax notoginseng and its extract have excellent antifungal and antioxidant activities, and thus, it is an important microbial resource for the developing novel drugs against plant fungal infections.

Gut microbiota and serum metabolome reveal the mechanism by which TCM polysaccharides alleviate salpingitis in laying hens challenged by bacteria.

This paper aimed to evaluate the effect of 3 kinds of TCM polysaccharides instead of antibiotics in preventing salpingitis in laying hens. After feeding the laying hens with Lotus leaf polysaccharide, Poria polysaccharide, and Epimedium polysaccharide, mixed bacteria (E. coli and Staphylococcus aureus) were used to infect the oviduct to establish an inflammation model. Changes in antioxidant, serum immunity, anti-inflammatory, gut microbiota, and serum metabolites were evaluated. The results showed that the 3 TCM polysaccharides could increase the expression of antioxidant markers SOD, GSH, and CAT, and reduce the accumulation of MDA in the liver; the contents of IgA and IgM in serum were increased. Decreased the mRNA expression of TLR4, NFκB, TNF-α, IFN-γ, IL1ß, IL6, and IL8, and increased the mRNA expression of anti-inflammatory factor IL5 in oviduct tissue. 16sRNA high-throughput sequencing revealed that the 3 TCM polysaccharides improved the intestinal flora disturbance caused by bacterial infection, increased the abundance of beneficial bacteria such as Bacteroides and Actinobacillus, and decreased the abundance of harmful bacteria such as Romboutsia, Turicibacter, and Streptococcus. Metabolomics showed that the 3 TCM polysaccharides could increase the content of metabolites such as 3-hydroxybutyric acid and isobutyl-L-carnitine, and these results could alleviate the further development of salpingitis. In conclusion, the present study has found that using TCM polysaccharides instead of antibiotics was a feasible way to prevent bacterial salpingitis in laying hens, which might make preventing this disease no longer an issue for breeding laying hens.

Functional characterization of group â ¡ interferon, IFNf in the acipenseriform fish, Chinese sturgeon (Acipenser sinensis).

Teleost fish possess a diversity of type Ⅰ interferons (IFNs) repertoire, which play a crucial role in antiviral and antimicrobial immune responses. In our previous study, IFNe1-3 and IFNb were identified and cloned from Chinese sturgeon (Acipenser sinensis), an acipenseriform fish. However, the absence of Chinese sturgeon genome data has left the question of whether there are other type Ⅰ IFN members in this species unresolved. In this study, we have identified and characterized a novel IFN, IFNf in Chinese sturgeon (AsIFNf). Bioinformatics analysis revealed that the AsIFNf contains a unique disulfide bond (2 cysteines) located in the second exon and fifth exon region, distinguishing it from other reported teleost type I IFNs. Meanwhile, qPCR results showed that AsIFNf mRNA was detectable in all examined tissues and up-regulated in the spleen or kidney in response to poly I: C, Citrobacter freundii, and Spring Viremia of Carp Virus (SVCV), but not by LPS. Furthermore, compared to recombinant AsIFNe2 protein (rAsIFNe2), rAsIFNf exhibited a stronger protective effect on Chinese sturgeon fin cells against SVCV and also induced higher expression of antiviral genes Mx and viperin. Importantly, AsIFNf displayed characteristics similar to antimicrobial peptides (AMPs) with a positive charge and demonstrated a broad spectrum of antimicrobial activity in vitro. These findings provide a theoretical foundation for understanding the primitive structure and function of interferon, as well as deepening our comprehension of the innate immune system and disease defense in the endangered Chinese sturgeon.

Rapid detection of grass carp reovirus type 1 using RPA-based test strips combined with CRISPR Cas13a system.

Introduction: Due to the existence of grass carp reovirus (GCRV), grass carp hemorrhagic disease occurs frequently, and its high pathogenicity and infectivity are great challenges to the aquaculture industry. As a highly pathogenic pathogen, the outbreak of hemorrhagic disease often causes tremendous economic losses. Therefore, it is important to rapidly and accurately detect GCRV on site to control timely. Methods: In this study, recombinant enzyme amplification (RPA) combined with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a system was employed to establish a method to detect the vp7 gene of grass carp reovirus type 1. This method can be adopted for judging the results by collecting fluorescence signal, ultraviolet excitation visual fluorescence and test strip. Results: Combined with the RPA amplification experiment, the detection limit of the RPA-CRISPR method can reach 7.2 × 101 copies/µL of vp7 gene per reaction, and the detection process can be completed within 1 h. In addition, this method had no cross-reaction with the other 11 common aquatic pathogens. Then, the performance of the RPA-CRISPR/Cas13a detection method was evaluated by comparing it with the real-time fluorescence quantitative PCR detection method of clinical samples. The results of RPA-CRISPR/Cas13a detection were shown to be in consistence with the results obtained from the real-time fluorescence quantitative PCR detection. The coincidence rate of this method with 26 GCRV clinical samples was 92.31%. Discussion: In summary, this method has high sensitivity, specificity and on-site practicability for detecting GCRV type 1, and has great application potential in on-site GCRV monitoring.

Diagnosis and treatment of venous thromboembolism and clinical application of inferior vena cava filter in China.

OBJECTIVE: This study investigated the trend of venous thromboembolism (VTE) in China during the past 10 years and assessed the clinical application of inferior vena cava filters (IVCFs). METHODS: A survey designed to investigate the diagnosis and management of VTE, specifically the application of IVCFs, was distributed nationally from January 2009 to December 2019. The respondents were mainly designated medical professionals and were asked to complete 4 major and 61 minor items in the survey. RESULTS: A total of 53 medical centers, including 27 radiologic and 26 vascular surgery centers, from 21 provinces in China participated in the study. These centers had diagnosed and treated 171,310 patients with VTE; 83,969 were inpatients (49%). During a 10-year period, an increasing trend of VTE diagnosis and inpatient management, 3.8-fold and 4.8-fold, respectively, was observed. The characteristics of the inpatients were as follows: 15% bilateral lower extremity deep vein thrombosis (DVT), 27% right lower extremity DVT, and 58% left lower extremity DVT. Anticoagulation therapy included unfractionated heparin with vitamin K antagonists (8%), low-molecular-weight heparin (LMWH) with vitamin K antagonists (21%), LMWH with transition to rivaroxaban (34.2%), LMWH with transition to dabigatran (2.4%), rivaroxaban alone (33.4%), and dabigatran alone (1.0%). The percentage of patients continuing anticoagulation therapy at 3, 6, 12, 24, and >24 months was 36%, 35%, 18%, 6.0%, and 5%, respectively. The in-hospital mortality for the patients with VTE was 3.2%, with DVT and pulmonary embolism responsible for 5.2% and DVT alone for 2.7%. Thrombolytic therapy was initiated for 39,046 of 83,969 patients (46.5%), including catheter-directed thrombolysis for 33,189 of the 39,046 patients (85%) and evaluation of the iliac vein using ultrasound and/or venography for 63,816 patients (76%). Urokinase was the main thrombolytic drug used (98%), followed by recombinant tissue-type plasminogen activator. Complete and partial thrombolysis was achieved in 70% and 30% of the patients, respectively. Bleeding complications were observed in 3.5% of patients, and 20% of the patients with bleeding complications required intervention. Between 2009 and 2019, 40,478 IVCFs (76% retrievable) were implanted in hospitalized VTE patients. During the enrollment period, the total number of IVCFs implanted increased by 3.8-fold, with a 4.8-fold increase in retrievable IVCFs and 7.5-fold decline in permanent IVCFs. The removal rate for the retrievable IVCFs was 72%. After IVCF implantation, 94.8% of patients received anticoagulation therapy for an average of 9.1 ± 8.6 months. The overall complication rate associated with IVCF placement was 15.5% (n = 6274 of 40,478 IVCFs), including tilting (54%), vena cava thrombosis (26.1%), caval penetration (12.6%), and migration (7.3%). No IVCF placement-related mortality occurred. CONCLUSIONS: A significant increase occurred in VTE diagnosis in China during the past decade. Anticoagulation therapy was the mainstay treatment, and catheter-directed thrombolysis was widely used. Most IVCFs placed were retrievable, and the use of permanent IVCFs has largely been discarded.

Adh Promotes Actinobacillus pleuropneumoniae Survival in Porcine Alveolar Macrophages by Inhibiting CHAC2-Mediated Respiratory Burst and Inflammatory Cytokine Expression.

Actinobacillus pleuropneumoniae (A. pleuropneumoniae) causes porcine pleuropneumonia that seriously endangers pig's health. Adh, located in the head region of trimeric autotransporter adhesion of A. pleuropneumoniae, affects bacterial adhesion and pathogenicity. However, how Adh mediates A. pleuropneumoniae immune invasion is still unclear. Here, we established the A. pleuropneumoniae strain L20 or L20 ΔAdh-infected porcine alveolar macrophages (PAM) model, and applied protein overexpression, RNA interference, qRT-PCR, Western blot and immunoflourescence techniques to dissect the effects of Adh on PAM during A. pleuropneumoniae infection. We found that Adh could increase the A. pleuropneumoniae adhesion and intracellular survival in PAM. Gene chip analysis of piglet lungs further showed that Adh significantly induced cation transport regulatory-like protein 2 (CHAC2) expression, whose overexpression suppressed the phagocytic capacity of PAM. Furthermore, CHAC2 overexpression dramatically increased glutathione (GSH) expression, decreased reactive oxygen species (ROS), and promoted A. pleuropneumoniae survival in PAM, while the knockdown of CHAC2 reversed these phenomena. Meanwhile, CHAC2 silence activated the NOD1/NF-κB pathway, resulting in an increase in IL-1ß, IL-6, and TNF-α expression, whereas this effect was weakened by CHAC2 overexpression and addition of NOD1/NF-κB inhibitor ML130. Moreover, Adh enhanced the secretion of LPS of A. pleuropneumoniae, which regulated the expression of CHAC2 via TLR4. In conclusion, through a LPS-TLR4-CHAC2 pathway, Adh inhibits respiratory burst and inflammatory cytokines expression to promote A. pleuropneumoniae survival in PAM. This finding may provide a novel target for the prevention and treatment of A. pleuropneumoniae.

The efficacy of stenting in the iliofemoral vein of patients with venous obstruction and secondary lymphedema from malignancy.

OBJECTIVE: To investigate the safety and effectiveness of venous stenting in patients with chronic iliofemoral venous obstruction and secondary lymphedema from malignancy. METHODS: From July 2012 to December 2020, patients with iliofemoral venous obstruction and secondary lymphedema who underwent venous stenting in our institution were reviewed retrospectively. Clinical characteristics, surgical complications, and symptom relief were assessed. Stent patency was evaluated with duplex ultrasound or computed tomographic venography. Twelve-month outcomes were reported. RESULTS: Fifty-three patients with concurrent secondary lymphedema who had stents placed for iliofemoral venous obstruction were included. There were 42 females, and the mean age was 56.9 years. Nonthrombotic iliac vein lesions were identified in 16 patients (30.1%). Immediate technical success was 100%, with an average of two stents implanted. The median Villalta score, and Chronic Venous Disease Quality of Life quality of life questionnaire scores decreased from 12 (IQR, 10-15) and 58 (IQR, 50-66) at baseline, respectively, to 5 (interquartile range [IQR], 4-6) and 28 (IQR, 22-45) at 12 months after the procedure (P < .05), showing significant improvement in the quality of life. At the end of a median follow-up of 12 months (range, 3-25 months), the cumulative primary, assisted primary, and secondary patency rates were 70.8%, 76.9%, and 90.1%, respectively. CONCLUSIONS: In patients with secondary lymphedema from malignancy, venous stent placement is safe and effective for iliofemoral venous obstruction.

The two-component system CpxA/CpxR is critical for full virulence in Actinobacillus pleuropneumoniae.

Actinobacillus pleuropneumoniae, a major bacterial porcine respiratory tract pathogen causing pig pleuropneumonia, has resulted in high economic losses worldwide. The mutation of the two-component system CpxAR strongly impacted the virulence of A. pleuropneumoniae, but the underlying regulatory mechanism remained unclear. Here, we found that CpxAR positively regulated the cpxDCBA gene cluster involved in polysaccharide capsule export. A capsular layer was confirmed in wild-type cells by transmission electron microscopy, whereas cpxAR and cpxD mutants were non-capsulated. The mutants for polysaccharide capsule export gene cpxD exhibited non-capsulated and were strongly impaired in virulence for mice, indicating a major role of CPS export system in virulence. We then demonstrated that CpxR directly regulated the transcription of the CPS export gene cluster cpxDCBA. Taken together, our data suggested that CpxAR is a key modulator of capsule export that facilitates A. pleuropneumoniae survival in the host.

Long noncoding RNA TUG1 induces angiogenesis of endothelial progenitor cells and dissolution of deep vein thrombosis.

OBJECTIVE: Long non-coding RNA (lncRNA) essentially controls many physiological and pathological processes of deep vein thrombosis (DVT). Based on that, lncRNA taurine upregulated gene 1 (TUG1)-involved angiogenesis of endothelial progenitor cells (EPCs) and dissolution of DVT was explored. METHODS: In the in-vitro experiments, EPCs were engineered with mimic, inhibitor, siRNA, and plasmid, after which tube formation, proliferation, migration, and apoptosis were checked. In the in-vivo experiments, a DVT mouse model was established. Before the DVT operation, the mice were injected with agomir, antagomir, siRNA, and plasmid. Subsequently, thrombosis and damage to the femoral vein were pathologically evaluated. TUG1, miR-92a-3p, and 3-Hydroxy-3-methylglutaryl coenzyme A reductase (Hmgcr) expression in the femoral vein was tested. The relationship between TUG1, miR-92a-3p, and Hmgcr was validated. RESULTS: DVT mice showed suppressed TUG1 and Hmgcr expression, and elevated miR-92a-3p expression. In EPCs, TUG1 overexpression or miR-92a-3p inhibition promoted cellular angiogenesis, whereas Hmgcr silencing blocked cellular angiogenesis. In DVT mice, elevated TUG1 or inhibited miR-92a-3p suppressed thrombosis and damage to the femoral vein whilst Hmgcr knockdown acted oppositely. In both cellular and animal models, TUG1 overexpression-induced effects could be mitigated by miR-92a-3p up-regulation. Mechanically, TUG1 interacted with miR-92a-3p to regulate Hmgcr expression. CONCLUSION: Evidently, TUG1 promotes the angiogenesis of EPCs and dissolution of DVT via the interplay with miR-92a-3p and Hmgcr.

Molecular Characterization of Outer Capsid Proteins VP5 and VP7 of Grass Carp Reovirus.

Aquareovirus, which is a member of the Reoviridae family, was isolated from aquatic animals. A close molecular evolutionary relationship between aquareoviruses and mammalian orthoreoviruses was revealed. However, the functions of the aquareovirus genome-encoded proteins are poorly understood. We investigated the molecular characteristics of the outer capsid proteins, namely, VP5 and VP7, of grass carp reovirus (GCRV). The peptides VP5 and VP7 were determined using in-gel tryptic digestion and mass spectrometry. Recovered peptides represented 76% and 66% of the full-length VP5 and VP7 sequences, respectively. Significantly, two-lysine acetylation, as well as two-serine and two-threonine phosphorylation modifications, were first revealed in VP5. We found that the initial amino acid in VP5 was Pro43, suggesting that a lower amount of VP5 remained uncleaved in virions at the autocleavage site (Asn42-Pro43). Further biochemical evidence showed that the cleaved VP5N/VP5C conformation was the major constituent of the particles. Moreover, early cleavage fragments of VP7 and enhanced infectivity were detected after limited tryptic digestion of GCRV, indicating that stepwise VP7 cleavage is essential for VP5 conformational rearrangement. Our results provide insights into the roles of posttranslational modifications in VP5 and its association with VP7 in the viral life cycle.

MicroRNA-136-5p from Endothelial Progenitor Cells-released Extracellular Vesicles Mediates TXNIP to Promote the Dissolution of Deep Venous Thrombosis.

OBJECTIVE: Endothelial progenitor cells-released extracellular vesicles (EPCs-EVs) have previously been reported to promote the dissolution of deep venous thrombosis (DVT) through delivery of microRNA (miR). Given that, this research was projected to search the relative action of EPCs-EVs transferring of miR-136-5p in DVT. METHODS: From EPCs transfected with miR-136-5p agomir or antagomir, EVs were extracted and then injected into DVT mice. Meanwhile, based on the treatment with EPCs-EVs loading miR-136-5p antagomir, silenced thioredoxin-interacting protein (TXNIP) lentivirus was injected into DVT mice to perform the rescue experiments. Afterwards, the length and weight of venous thrombosis, EPC apoptosis and inflammatory factors, plasmin, fibrinogen, and thrombin-antithrombin were measured. miR-136-5p and TXNIP expression in DVT mice, and their targeting relationship were evaluated. RESULTS: miR-136-5p expression was suppressed and TXNIP expression was elevated in DVT mice. EPCs-EV reduced the length and weight of venous thrombosis, suppressed cell apoptosis and inflammatory reaction, as well as elevated level of plasmin, and reduced levels of fibrinogen and thrombin-antithrombin in DVT mice. Restored miR-136-5p loaded by EPCs-EV further attenuated DVT but EPCs-EV transfer of depleted miR-136-5p resulted in the opposite consequences. miR-136-5p targeted TXNIP and silenced TXNIP rescued the effect of EPCs-EV transfer of depleted miR-136-5p on DVT. CONCLUSION: miR-136-5p from EPCs-EV suppresses TXNIP expression to reduce the thrombus size in DVT, offering a promising treatment target for DVT.

A case report of CAT gene and HNF1ß gene variations in a patient with early-onset diabetes.

Complex forms of diabetes are the ultimate common pathway involving multiple genetic variations and multiple environmental factors. Type 2 diabetes (T2DM) is classified as complex diabetes. Varying degrees of insulin deficiency and tissue insulin resistance are two key links to T2DM. The islet ß cell dysfunction plays a crucial role in the pathogenesis of T2DM. The decompensation of the islet ß cell to insulin resistance is a common mechanism leading to the pathogenesis of T2DM. Available data show that genetic factors mainly affect cell function. At present, a number of susceptibility genes related to T2DM have been reported at home and abroad. In this study, the diabetes-related genes in the case of early-onset diabetes with a significant family history were examined, and our results showed the presence of the intron mutations of catalase (CAT) gene and hepatocyte nuclear factor 1ß (HNF1ß) gene. The patient enrolled in this study was observed and analyzed, thus, increasing further understanding of the genes associated with diabetes and exploring the pathogenesis of diabetes from the molecular level. This is significant for guiding the prevention, treatment, and prognosis evaluation of diabetes.

Anti-coagulation and anti-hyperplasia coating for retrievable vena cava filters by electrospraying and their performance in vivo.

A novel drug eluting retrievable vena cava filter (RVCF) with a heparin-modified poly(ε-caprolactone) (hPCL) coating containing rapamycin was prepared by electrospraying. The in vitro drug release pattern showed that the encapsulated rapamycin in the coating can be sustainably released within one month, whereas activated partial thromboplastin time (APTT) and in vitro cell culture showed that the drug eluting RVCF can effectively extend blood clotting time and inhibit smooth muscle cell (SMC) and endothelial cell (EC) proliferation, respectively. The as-prepared drug eluting RVCF and corresponding commercial RVCF were implanted into the vena cava of sheep. The retrieval operation at a predetermined time point showed that the drug eluting RVCF had a much higher retrieval rate than the commercial RVCF. Comprehensive investigations, including histological, immunohistological and immunofluorescence analyses, on explanted veins were carried out. The results demonstrated that the as-prepared RVCF possessed excellent antihyperplasia properties in vivo, significantly improving the retrieval rate and extending the in vivo dwelling time in sheep. Consequently, the drug eluting RVCF has promising potential for application in the clinic to improve RVCF retrieval rates.

The CpxA/CpxR two-component system mediates regulation of Actinobacillus pleuropneumoniae cold growth.

Introduction: To survive in various hostile environments, two-component system is an adaptive mechanism for diverse bacteria. Activity of the CpxA/CpxR two-component system contributes to coping with different stimuli, such as pH, osmotic and heat stress. Methods: However, the role of the CpxA/CpxR system in cold resistance is little-known. In this study, we showed that CpxA/CpxRwas critical for A. pleuropneumoniae growth under cold stress. Results: ß-Galactosidaseanalysis showed that CpxA/CpxR positively regulated the predicted cold stress gene cspC. The mutant for cold stress gene cspC was impaired in the optimal growth of A. pleuropneumoniae under cold stress. Furthermore, electrophoretic mobility shift assays demonstrated that CpxR-P could directly regulate the transcription of the cold stress gene cspC. Discussion: These results presented in this study illustrated that the CpxA/CpxR system plays an important role in cold resistance by upregulating expression of CspC. The data give new insights into how A. pleuropneumoniae survives in cold stress.

Ultrasound-Guided Retrograde Infrapopliteal Artery Access for Recanalization of Complex Femoral-Popliteal Artery Occlusions.

BACKGROUND: The aim of the study was to review the outcomes of femoral-popliteal artery (FPA) interventions using an ultrasound (US)-guided retrograde infrapopliteal artery access after the failure of an antegrade recanalization. METHODS: From Jan 2016 to Jan 2019, 37 patients with chronic total occlusion (CTO) of the FPA underwent ultrasound (US)-guided retrograde infrapopliteal artery access after failure of an antegrade procedure. Treated limbs were classified as Rutherford class 5 or 6 (29.7%) and class 4 (62.2%). Data collected included success rate and time to access using US. Immediate in-hospital and follow-up outcomes were also documented. RESULTS: US-guided retrograde infrapopliteal artery access was successful in 100% of the patients (anterior tibial = 11, posterior tibial = 19, Peroneal = 4, Dorsalis pedis = 3). Retrograde revascularization was achieved in all 37 patients (100%) using balloon angioplasty (17/37, 45.9%) and additional stent placement (20/37, 54.1%). Ankle-brachial index (ABI) measurements changed from 0.25 ± 0.1 preinterventionally to 0.75 ± 0.07 at 1 day postinterventionally (<0.001). Minor complications occurred in 2/37 patients (5.4%) including one bleeding and vasospasm at the posterior tibial artery, both of which were treated conservatively. No patient experienced access-related thrombosis, aneurysm, compartment syndrome or death. Thirty of 37 (81%) patients completed for at least 12 months of follow-up. None of the successful revascularized patients had major or minor amputations during the follow-up period. CONCLUSIONS: US-guided retrograde infrapopliteal artery access is a safe and successful technique, which expands revascularization options after the failure of conventional endovascular antegrade approaches.

Plasma proteomic screening and validation of novel biomarkers in Takayasu's arteritis.

OBJECTIVES: To screen and validate differential proteins as novel biomarkers in active Takayasu's arteritis (TAK). METHODS: Plasma samples from 40 active, 40 inactive patients, and 40 healthy controls were collected. Protein profiles of plasma were mapped by two-dimensional gel electrophoresis. Differential protein spots were detected and identified by image analysis and mass spectrometry. Plasma concentrations of proteins were measured to validate candidate biomarkers. The area under the receiver operating characteristic (ROC) curve (AUC) of circulating plasma concentrations of candidate biomarkers were calculated to assess diagnostic value. RESULTS: With a total of 1507 matched gel spots, there were 170 differential expression spots between active and inactive TAK, including 139 up-regulated and 31 downregulated. Only 11 proteins could be identified by mass spectrometry. Serum amyloid A(SAA), fibrinogen, complement C4a, complement C3c, complement C4b binding protein(C4bp), recombination acting gene protein 1(RAG1), alpha-1-acid glycoprotein, alpha-1-microglobulin, complement C7, complement factor H related protein-1 were up-regulated in active patients, while serum amyloid P was down-regulated. Active patients had higher circulating levels of RAG1(P<0.001), C4bp (p=0.012) and SAA (p<0.001), compared to inactive patients, while inactive patients had higher levels than controls (RAG1, p=0.011; C4bp, p=0.012; SAA, p=0.005). The composite AUC with SAA, RAG1, and C4bp was 0.94 (95%CI 0.86-0.98) for discriminating activity, larger than 0.71(95% CI 0.60-0.80) for ESR (p=0.0004) or 0.75(95%CI 0.64-0.84) for CRP (p=0.0014), respectively. ONCLUSIONS: Some acute-phase and immunology-related proteins may serve as novel biomarkers of TAK. Further study of these proteins may be helpful to elucidate the pathologic mechanism.

Clinical outcomes of radiofrequency ablation for patients with varicose veins of the lower extremities combined with grade II iliac vein compression.

BACKGROUND: Iliac vein compression (IVC) is a common condition in patients with varicose veins (VVs) of the legs. IVC has been classified into three grades in previous studies. Grade II IVC is defined by >50% stenosis without the development of collateral circulation. The purpose of the present study was to investigate the outcomes of radiofrequency ablation (RFA) for patients with VVs combined with grade II IVC. METHODS: A retrospective analysis was conducted of 339 patients who had undergone RFA for VVs of the left leg from March 2017 to January 2019. Duplex ultrasonography, computed tomography venography, and venography were performed to evaluate for grade II IVC. All the patients were divided into two groups. Group 1 included patients with VVs only, and group 2, patients with VVs combined with grade II IVC. Propensity score matching was used to ensure an even distribution of confounding factors between groups. The venous clinical severity score (VCSS) and chronic venous insufficiency questionnaire (CIVIQ) score were recorded during the 12-month follow-up. Occlusion of the truncal veins was evaluated using duplex ultrasound scans. RESULTS: Using 1:1 propensity score matching, 50 pairs of patients were enrolled in the present analysis. The average age of groups 1 and 2 was 58.7 ± 13.1 and 60.1 ± 7.1 years, respectively. The VCSS had decreased significantly from baseline to 12 months postoperatively (group 1, from 5 to 1; group 2, from 4 to 1; P < .01). A significant increase in the CIVIQ score was found between the baseline and 12-month evaluations for both groups (group 1, from 62.5 to 69; group 2, from 63 to 70; P < .01). The truncal occlusion rate was 98% in both groups at 12 months. No significant differences were found between the two groups in the VCSS, CIVIQ score, procedure complications, or occlusion rate during the 12-month follow-up. CONCLUSIONS: RFA is effective for patients with VVs combined with grade II IVC.

Development of a Magnetic Kissing Guidewire and Animal Experiment: a Preliminary Study.

Endovascular therapy has been the first option for many vascular diseases. Due to the increasing development of endovascular devices and techniques in recent decades, endovascular procedures could be applied in treating those cases with complex anatomy. Two or three vascular access routes are required to establish through-and-through access in endovascular therapy for arterial occlusive disease and complex aortic aneurysm. Guidewire snaring with a snare kit is essential but sometimes time-consuming in the establishment of through-and-through access. To simplify the procedure, we design the magnetic kissing guidewire (MKG) with a magnetic tip that attracts each other to establish a guidewire route that meets the needs of clinical practice. We conducted the in vitro test to evaluate its magnetic force and the in vivo test to assess its performance in the arteries of twelve sheep. The results revealed that this novel guidewire significantly simplified the establishment of through-and-through access.