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 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.

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.

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.

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.

Structure of RNA polymerase complex and genome within a dsRNA virus provides insights into the mechanisms of transcription and assembly.

Most double-stranded RNA (dsRNA) viruses transcribe RNA plus strands within a common innermost capsid shell. This process requires coordinated efforts by RNA-dependent RNA polymerase (RdRp) together with other capsid proteins and genomic RNA. Here we report the near-atomic resolution structure of the RdRp protein VP2 in complex with its cofactor protein VP4 and genomic RNA within an aquareovirus capsid using 200-kV cryoelectron microscopy and symmetry-mismatch reconstruction. The structure of these capsid proteins enabled us to observe the elaborate nonicosahedral structure within the double-layered icosahedral capsid. Our structure shows that the RdRp complex is anchored at the inner surface of the capsid shell and interacts with genomic dsRNA and four of the five asymmetrically arranged N termini of the capsid shell proteins under the fivefold axis, implying roles for these N termini in virus assembly. The binding site of the RNA end at VP2 is different from the RNA cap binding site identified in the crystal structure of orthoreovirus RdRp λ3, although the structures of VP2 and λ3 are almost identical. A loop, which was thought to separate the RNA template and transcript, interacts with an apical domain of the capsid shell protein, suggesting a mechanism for regulating RdRp replication and transcription. A conserved nucleoside triphosphate binding site was localized in our RdRp cofactor protein VP4 structure, and interactions between the VP4 and the genomic RNA were identified.

Effectiveness and long-term outcomes of different crossing strategies for the endovascular treatment of iliac artery chronic Total occlusions.

BACKGROUND: The iliac occlusive disease is usually treated with endovascular procedures in recent years. The effectiveness of different crossing approaches for these occlusions is not precisely known. We performed a retrospective study to explore the optimal crossing approach (antegrade versus retrograde) for iliac artery chronic total occlusions (CTOs) and to examine the long-term outcomes. MATERIALS AND METHODS: We performed a study on 107 patients (116 iliac occlusive lesions, mean age 64.0 ± 11.1, 88 men) who underwent an iliac CTO endovascular intervention attempted with the use of both crossing strategies but were managed with one final crossing approach between August 2012 and August 2018. Baseline data, procedural characteristics, and outcomes were described. A Cox proportional hazard model and Kaplan-Meier method were developed to assess the differences in the two crossing approaches in terms of the 1-year and 5-year primary patency rates, target lesion revascularization (TLR) and major adverse limb events (MALEs). RESULTS: Common iliac artery (CIA) lesions were more likely to be crossed successfully in the retrograde direction (6.8% for antegrade vs. 20.9% for retrograde, p = 0.005), while lesions in the CIA/ external iliac artery (EIA) were more prone to be crossed successfully in the antegrade direction (58.9% for antegrade vs. 39.5% for retrograde, p = 0.016). There were no significant differences in the crossing approach for EIA lesions between the two groups. The two crossing approaches were associated with similar estimates of 1- and 5-year primary patency, TLR and MALE rates. CONCLUSION: The antegrade approach was associated with a higher rate of successful crossing in CIA/EIA CTO lesions, while the CIA-only CTOs were more likely to be crossed successfully with the retrograde approach.

A Randomized, Double-Blind, Placebo-Controlled Phase II Study of Hepatocyte Growth Factor in the Treatment of Critical Limb Ischemia.

NL003 is a plasmid engineered to simultaneously express two isoforms of hepatocyte growth factor. This phase II study was performed to assess the clinical safety and efficacy of intramuscular injection of NL003 in critical limb ischemia (CLI) patients for 6 months. Two hundred patients (Rutherford scale 4-5) were randomly assigned: placebo (n = 50), low-dose NL003 (n = 50), middle-dose NL003 (n = 50), or high-dose NL003 (n = 50). The drug was administered in the affected limb of 197 patients on days 0, 14, and 28. No significant differences in the incidence of adverse events (AEs) or serious AEs were found among the groups. At 6 months, pain severity was significantly reduced in all NL003 groups, but not in the placebo group (p < 0.05). The proportion of patients with complete ulcer healing in the high-dose group was significantly higher than that of the placebo group (p = 0.0095). There were no statistically significant differences in transcutaneous oxygen pressure (TcPO2), ankle-brachial index (ABI), or toe-brachial index (TBI) value among the four groups throughout the study period. These results provide the first effective evidence of significant improvements in total healing of ulcers in treated legs, complete pain relief without analgesics, and safety for NL003 in patients with Rutherford stage 4-5.

Complete Laparoscopic Retrieval of Inferior Vena Cava Filter: A Case Report and Literature Review.

BACKGROUND: In recent years, retrievable inferior vena cava filters (IVCFs) have been increasingly used to prevent pulmonary embolism. However, these IVCFs are occasionally difficult to retrieve, and their long-term retention in the body can cause various complications. METHODS: A 22-year-old woman underwent a prophylactic IVCF placement for puerperal deep venous thrombosis 6 months ago. After several attempts, the filter could not be retrieved through the endovascular technique. Meanwhile, the patient developed severe depression and suicidal tendencies. We decided to use laparoscopic techniques to retrieve the filter. RESULTS: We successfully used laparoscopy to completely remove a difficult IVCF. The patient had a smooth postoperative recovery and was discharged on the third postoperative day. CONCLUSIONS: Laparoscopy can be used as an alternative method to remove IVCF. However, it is more important to avoid situations that prevent routine retrieval of IVCFs.

Varicose Veins of the Lower Extremity Secondary to Tricuspid Regurgitation.

Pulsating varicose veins are a rare clinical finding and are mainly derived from tricuspid regurgitation or right heart failure. The precise causes and optimal treatment of this phenomenon have been poorly recorded in the literature. Here, we describe a 56-year-old woman who presented to our medical center with bilateral varicose veins, heaviness, and edema in her lower limbs. The duplex revealed an arterial-like pulsating flow in the superficial and deep veins of the lower extremities, in addition to severe tricuspid regurgitation. Symptoms improved after the patient was given compression therapy using elastic stockings. In this article, we also review six other cases from the literature and discuss the therapies that would be reasonable in some conditions.

Preparation and experimental research into retrievable rapamycin- and heparin-coated vena cava filters: a pilot study.

The use of retrievable vena cava filters (RVCFs) was once commonplace, but filter retrieval was often very difficult. Most unsuccessful retrieval was due to intimal hyperplasia of the inferior vena cava and in-filter thrombosis. This pilot study aimed to design a drug-eluting RVCF. The hypothesis was that coated drugs could be released continuously to inhibit vena intimal hyperplasia and thrombosis, and thus improve the retrieval rates of RVCFs. Various concentrations of polycaprolactone (PCL)/chloroform solution were made from a mixture of Rapamycin and Heparin according to the quality of PCL. The drug was coated onto the surface of the filters by a process of dipping. In vitro tests were performed to check stability and in vitro drug release. Animals receiving filter implantation were divided into 4 groups, the experimental intervention group (EI), experimental laparotomy group (EL), control intervention group (CI), and control laparotomy group (CL). Filters were retrieved by laparotomy in the EL and CL groups, and by interventional operation in the EI and CI groups at 10, 20 and 30 days after implantation. Pathological endothelia biopsies were performed with wood grain-eosin (HE) staining and immunohistochemical examination, with the proliferating cell nuclear antigen (PCNA) index, and the results were compared between the experimental and control groups. The weight of thrombus within the filters was also measured by scale and compared. The coating concentration that succeeded in completely covering the surface was 0.2 g/ml. There was better coverage by SEM at this concentration, and the coated drugs had no obvious loss after filter release. The drug release curves showed that the amount of Heparin released was more than 50 % at day 1; Rapamycin released little in the first few days, beginning in earnest at 20 to 30 days. The filters were easy to retrieve at 10 days for both groups, while neither could be retrieved at 30 days. However, at 20 days the filter in the EI group could be retrieved with some difficulty, but the filter in the CI group couldn't be removed at all. The pathological examination and immunohistochemical PCNA examination results showed that the use of drug-eluting filters could effectively inhibit endothelial hyperplasia at 10 and 20 days, but was less effective at 30 days. There was no apparent difference in the total weight of blood clots between the experimental and control groups. We successfully conducted a pilot study into preparing Rapamycin- and Heparin-coated RVCFs. In vitro and in vivo tests further proved the possibility of improving the retrieval rates of RVCFs by effectively inhibiting vein endothelial proliferation, but the anticoagulation and antithrombosis effects of Heparin were unsatisfactory.

VP5 autocleavage is required for efficient infection by in vitro-recoated aquareovirus particles.

Grass carp reovirus (GCRV) is a member of the genus Aquareovirus in the family Reoviridae, and contains five core proteins (VP1-VP4 and VP6) and two outer-capsid proteins (VP5 and VP7) in its particle. Previous studies have revealed that the outer-capsid proteins of reovirus are responsible for initiating infection, but the mechanism is poorly understood. Using baculovirus-expressed VP5 and VP7 to recoat purified cores, in vitro assembly of GCRV was achieved in this study. Recoated GCRV (R-GCRV) closely resembled native GCRV (N-GCRV) in particle morphology, protein composition and infectivity. Similar to N-GCRV, the infectivity of R-GCRV could be inhibited by treating cells with the weak base NH4Cl. In addition, recoated particles carrying an Asn→Ala substitution at residue 42 of VP5 (VP5N42A/VP7 R-GCRV) were no longer infectious. These results provide strong evidence that autocleavage of VP5 is critical for aquareovirus to initiate efficient infection.

Identification of a functional motif in the AqRV NS26 protein required for enhancing the fusogenic activity of FAST protein NS16.

Aquareoviruses AqRVs have a close relationship with orthoreoviruses. However, they contain an additional genome segment, S11, which encodes nonstructural protein NS26. We previously showed that NS26 can enhance the fusogenic activity of the fusion-associated small transmembrane FAST protein NS16 from AqRV. In this study, a TLPK motif in NS26 was identified as being important for the enhancement. When the TLPK motif was deleted from NS26, the enhanced efficiency of the NS16-mediated cellcell fusion was significantly impaired. Further mutational analysis showed that the lysine K residue in the TLPK motif was critical for the enhancement. Additionally, deletion of the TLPK motif prevented NS26 from interacting with lysosomes. These findings suggested that the TLPK motif is important for NS26 to enhance the fusogenic activity of NS16, and NS26 may utilize the lysosome to benefit the fusion process.

[Recent progress in application of local drug delivery for preventing restenosis after peripheral arterial revascularization].

Restenosis following vascular revascularization remains an important clinical problem. Local drug delivery which can provide enough drug concentration in the lesion location without causing adverse systemic effect is an excellent solution for this question. We conducted a systematic literatory search on PubMed and CKNI through May 2014. After reviewing all related papers, we provided a comprehensive overview of the available drugs and techniques for local drug delivery that have been developed to prevent restenosis after peripheral vascular interventions, including innovations that have been tested only in animals as well as those already approved for clinical use. In brief, anti-proliferative drugs such as paclitaxel and sirolimus are the most used and suitable drugs for local delivery system. Additionally, some promising drugs including anti-inflammatory drugs, antioxidant drugs and drugs inhibiting cell proliferation and migration are already being tested in pre-clinical trials or animal models. At the same time, intraluminal and extraluminal delivery devices have also got a rapid development during the past decades. The efficacy of drug-eluting stent, drug-eluting balloon, porous and microporous balloon and the most recent drug-eluting bioresobable scaffold for preventing of restenosis in peripheral vessels have been demonstrated in humans or in animals, some of them even have received the CE mark in Europe. Endovascular microinfusion catheter and drug-loaded perivascular wraps have only been tested in animal models, more researches are needed. With the development of pharmacology and bioengineering, great strides will be made in the prevention of restenosis in the near future.

[Risk factors associated with the severity of pulmonary embolism in patients with acute deep venous thrombosis of lower extremities].

OBJECTIVE: To identify the risk factors associated with the severity of pulmonary embolism among patients with deep venous thrombosis of lower extremities. METHODS: This prospective study enrolled 208 patients with acute deep venous thrombosis to screen for pulmonary embolism between July 2010 and July 2012 in Beijing Shijitan Hospital. There were 101 male and 107 female patients, with a mean age of (59 ± 16) years. Gender, age, extension, side of lower extremities of deep venous thrombosis was analyzed by χ² test. Ordinal Logistic regression was used to determine risk factors associated with severity of pulmonary embolism. RESULTS: There were 83 patients with iliofemoral deep venous thrombosis, 102 patients with femoropopliteal and 23 patients with calf deep venous thrombosis. Pulmonary embolism was detected in 70 patients with the incidence of 33.7%. Pulmonary embolism was significantly correlated with extension (χ² = 17.286, P = 0.004) and sides (χ² = 15.602, P = 0.008) of deep venous thrombosis, not with age (χ² = 7.099, P = 0.260), gender (χ² = 7.014, P = 0.067), thrombotic risk factors (χ² = 3.335, P = 0.345) in univariate analysis. Results of multivariate ordinal logistic regression showed that iliofemoral vein thrombosis (OR = 6.172, 95% CI: 1.590 to 23.975, P = 0.009) and bilateral venous thrombosis (OR = 7.140, 95% CI: 2.406 to 24.730, P = 0.001) are associated with more serious pulmonary embolism. CONCLUSIONS: Incidence of pulmonary embolism is still high in patients with deep venous thrombosis. Extensive iliofemoral and bilateral vein thrombosis may increase risk of severity of pulmonary embolism. Clinicians should pay more attention to these high-risk patients.

Catheter-directed thrombolysis-assisted angioplasty for chronic lower limb ischemia.

BACKGROUND: Thrombolysis is an appropriate treatment for acute arterial occlusion. There remains controversy as to whether thrombolysis before angioplasty helps to identify the underlying lesion and improve results for chronic ischemia of the lower extremity. We sought to investigate the feasibility of catheter-directed thrombolysis-assisted angioplasty for chronic lower limb ischemia. METHODS: Between July 2008 and December 2009, the data of patients with chronic lower limb ischemia undergoing catheter-directed thrombolysis-assisted angioplasty were retrospectively analyzed. RESULTS: Twenty consecutive patients (18 men with a mean age of 56.35 ± 8.5 years) underwent thrombolysis-assisted angioplasty for occlusion of a native artery (n = 18) or bypass graft (n = 2). The median duration of symptoms was 19 months (range: 3-48 months). Symptoms included disabling claudication in 12 patients, rest pain in 5 patients, and gangrene of the toes in 3 patients. Urokinase or recombinant tissue plasminogen activator as a thrombolytic agent was used before angioplasty. The mean length of occlusive lesions decreased significantly from 150 mm to 30 mm after thrombolysis (P < 0.01). Four patients had no change in their lesions. Improvement of Inter-Society Consensus for the Management of Peripheral Arterial Disease (TASC II) classification was achieved in 16 patients, with 14 TASC IIA lesions and 2 TASC IIB lesions after thrombolysis. Subsequent stenting was successfully performed in all patients. The ankle-brachial index increased significantly from 0.33 to 0.63 (P < 0.01). No perioperative deaths occurred. Morbidity included access site bleeding in 1 patient and distal embolization in 2 patients without further intervention. The primary patency rate at 1 year was 95%, with a median follow-up time of 19 months. CONCLUSIONS: Catheter-directed thrombolysis-assisted angioplasty is a safe and effective treatment in some patients with chronic lower limb ischemia. It may reduce the magnitude of the lesion and simplify the expected intervention procedures.

[Exercise versus immobilization in the treatment of acute deep vein thrombosis during different clot-organized stage: an animal experiment].

OBJECTIVE: To evaluate the corresponding influence on pulmonary embolism incidence between immobilization and exercise in different stage of thrombus after acute deep vein thrombosis in rabbits. METHODS: Forty-eight New Zealand rabbits were randomly divided into three groups depending on the different organized stage of thrombus: the early, medium and later stage group.Each group was subdivided into two sub groups: the immobile and mobile subgroup. Rabbit modeling of deep vein thrombosis was made by ligating the right femoral vein. Among the early-stage group, rabbits of the immobile subgroup were fixed for 3 days, while that of the mobile subgroup were free to move for 3 days, then each was euthanized to extract the lungs for pathological examination. Among the medium-stage group, each of the immobile subgroup were fixed for 7 days, while the mobile subgroup ones were fixed for 3 days, then released free-moving for 4 days following the pathological extraction. Among the later-stage group, animals in the immobile subgroup were fixed for 14 days comparing the mobile subgroup fixed for 7 days and next free-moving for 7 days, then each was euthanized. RESULTS: Among the early-stage group, pulmonary embolism incidence (PEI) of the immobile and mobile subgroup was 4/8 vs.3/8, the pulmonary lobe embolism incidence (PLEI) was 17.5% (7/40) vs. 15.0% (6/40). Among the medium-stage group, PEI of the immobile and mobile subgroup was 3/8 vs. 2/8, PLEI was 37.5% (7/40) vs. 25.0% (10/40). Among the later-stage group, PEI of the immobile and mobile subgroup was 3/8 vs. 3/8, PLEI was 12.5% (5/40) vs. 15.0% (6/40). There was no statistical difference between immobilization subgroup and mobilization subgroup among different stage group. CONCLUSION: On the premise of given anticoagulation treatment, early ambulation do not significantly increase pulmonary embolism incidence after acute deep vein thrombosis of lower extremity in rabbits.

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.

A prospective randomized controlled clinical trial on clopidogrel combined with warfarin versus clopidogrel alone in the prevention of restenosis after endovascular treatment of the femoropopliteal artery.

BACKGROUND: We sought to compare the effects of clopidogrel combined with warfarin with clopidogrel alone in the prevention of restenosis after endovascular treatment (EVT) of the femoropopliteal artery. METHODS: Between June 2008 and May 2009, 88 consecutive patients referred for EVT were randomly divided into a clopidogrel group (42 cases) and a clopidogrel combined with warfarin group (46 cases) before the procedure. Examinations including staging of peripheral arterial disease by Rutherford, ankle-brachial index, and color duplex ultrasonography were performed at baseline, 1 week, 3 months, 6 months, and 12 months after procedure. At the same time, bleeding complications were observed. RESULTS: Fifty patients (63 limbs) were included after 12 months of follow-up, in which 25 patients (30 limbs) were from the clopidogrel group and 25 patients (33 limbs) were from the combination group. At 3 months, the rates of restenosis on duplex ultrasonography were 17% in the clopidogrel group and 18% in the combination group (P = 1.0). At 6 months, the accumulated restenosis rates were 37% and 36% (P = 0.98), respectively. At 12 months, the accumulated restenosis rates were 53% and 42% (P = 0.523), respectively. The rate of clinical bleeding events was 21% (6/29) in the combination group compared with 7% (2/27) in the clopidogrel group, and there was no statistical difference (P = 0.3). CONCLUSIONS: The combination of clopidogrel with warfarin was not more effective than clopidogrel alone in restenosis prevention for patients who underwent EVT. Instead, the combination of antiplatelet and anticoagulation therapy was inclined to increase the clinical bleeding events.