In angiosperms, pollen wall pattern formation is determined by primexine deposition on the microspores. Here, we show that AUXIN RESPONSE FACTOR17 (ARF17) is essential for primexine formation and pollen development in Arabidopsis (Arabidopsis thaliana). The arf17 mutant exhibited a male-sterile phenotype with normal vegetative growth. ARF17 was expressed in microsporocytes and microgametophytes from meiosis to the bicellular microspore stage. Transmission electron microscopy analysis showed that primexine was absent in the arf17 mutant, which leads to pollen wall-patterning defects and pollen degradation. Callose deposition was also significantly reduced in the arf17 mutant, and the expression of CALLOSE SYNTHASE5 (CalS5), the major gene for callose biosynthesis, was approximately 10% that of the wild type. Chromatin immunoprecipitation and electrophoretic mobility shift assays showed that ARF17 can directly bind to the CalS5 promoter. As indicated by the expression of DR5-driven green fluorescent protein, which is an synthetic auxin response reporter, auxin signaling appeared to be specifically impaired in arf17 anthers. Taken together, our results suggest that ARF17 is essential for pollen wall patterning in Arabidopsis by modulating primexine formation at least partially through direct regulation of CalS5 gene expression.

dSevere fever with thrombocytopenia syndrome (SFTS) is a newly emerging and epidemic infectious disease in central and northeast China. It is caused by New Bunyavirus and carries an average 12% case fatality rate. Early and rapid detection is critical for prevention and control of New Bunyavirus infection, since no vaccine or antiviral drugs are currently available, and prevention requires careful attention to control of the suspected tick vector. In this study, a simple and sensitive reverse transcription-loopmediated isothermal amplification (RT-LAMP) assay was developed for rapid detection of New Bunyavirus. The detection limit of the RT-LAMP assay was approximately 10 3 50% tissue culture infective doses/ml of New Bunyavirus in culture supernatants, and no cross-reactive amplification of other viruses known to cause similar clinical manifestations was observed. The assay was further evaluated using 138 specimens from clinically suspected SFTS and 40 laboratory-proven hantavirus infection with fever and renal syndrome patients, and the assay exhibited 97% agreement compared to real-time RT-PCR and conventional RT-PCR. Using real-time RT-PCR as the diagnostic gold standard, RT-LAMP was 99% sensitive and 100% specific. The RT-LAMP assay could become a useful alternative in clinical diagnosis of SFTS caused by New Bunyavirus, especially in resource-limited hospitals or rural clinics of China. Since 2007, many patients in China have presented with an illness with clinical characteristics that include acute onset of fever with leukopenia, thrombocytopenia, elevated serum hepatic enzyme levels, gastrointestinal manifestations, neurological symptoms, and bleeding tendencies. The disease was initially suspected to be human granulocytic anaplasmosis (HGA) (1), but in 2009, difficulty in proving that etiology prompted syndromic characterization of the patients as having "severe fever with thrombocytopenia syndrome" (SFTS), while other potential etiologies were investigated (2). A novel bunyavirus was isolated from patient acute-phase serum samples and is now known as New Bunyavirus (NBV), severe fever with thrombocytopenia syndrome bunyavirus (SFTSV), or Huaiyangshan virus (HYSV) (1-3).As the clinical manifestations of SFTS are nonspecific, there is a need to develop assays that can differentiate this from various other infectious diseases, in particular, hemorrhagic fever with renal syndrome (HFRS) and HGA. Reliable rapid and accurate diagnostic tests are urgently required to identify and diagnose clinically suspected SFTS, to assess treatment efficacy, and for disease surveillance (4).Recently, a novel DNA amplification method called loop-mediated isothermal amplification (LAMP) has been applied for diagnosis of infectious diseases (5). The addition of reverse transcriptase makes it possible to amplify RNA sequences by LAMP (reverse transcription-loop-mediated isothermal amplification, or RT-LAMP). RT-LAMP has already been applied to the detection of several RNA viruses, such as enterovirus 71 (EV71), swineorigin inf...

Background: Sedum sarmentosum is traditionally used to treat various inflammatory diseases in China. It has protective effects against acute liver injury, but the exact mechanism of such effects remains unclear. This study investigated the protective effects of S. sarmentosum extract on lipopolysaccharide (LPS)/D-galactosamine (D-GalN)induced acute liver injury in mice and the mechanism of such effects.Methods: Mice were randomly divided into control, treatment, model, and model treatment groups. Acute liver injury was induced in model mice via intraperitoneal injection of LPS and D-GalN with doses of 10 μg/kg of LPS and 500 mg/kg, respectively. The mRNA expression levels of miR-124, Hedgehog, Patched (Ptch), Smoothened (Smo), and glioma-associated oncogene homolog (Gli) in liver tissues were determined through RT-PCR, and the protein levels of Hedgehog, Ptch, Smo, Gli, P13k, Akt, HMGB1, TLR4, IkB-α, p-IkB-α, and NF-kB65 were evaluated via Western blot analysis. The serum levels of IL-6, TNF-α, CRP, IL-12, and ICAM-1 were determined via ELISA. TLR4 and NF-κBp65 activity and the levels of DNA-bound NF-KB65 and TLR4 in LPS/D-GalN-induced liver tissues were also determined. We recorded the time of death, plotted the survival curve, and calculated the liver index. We then observed the pathological changes in liver tissue and detected the levels of liver enzymes (alanine aminotransferase [ALT] and aspartate transaminase [AST]) in the serum and myeloperoxidase (MPO) and plasma inflammatory factors in the liver homogenate. Afterward, we evaluated the protective effects of S. sarmentosum extracts on acute liver injury in mice.Results: Results showed that after S. sarmentosum extract was administered, the expression level of miR-124 increased in liver tissues. However, the protein expression levels of Hedgehog, Ptch, Smo, Gli, P13k, p-Akt, HMGB1, TLR4, p-IκB-α, and NF-κB65 and the mRNA expression levels of Hedgehog, Ptch, Smo, and Gli decreased. The MPO level in the liver, the IL-6, TNF-α, CRP, IL-12, and MMP-9 levels in the plasma, and the serum ALT and AST levels also decreased, thereby reducing LPS/D-GalN-induced liver injury and improving the survival rate of liver-damaged animals within 24 h.Conclusions: S. sarmentosum extract can alleviate LPS/D-GalN-induced acute liver injury in mice and improve the survival rate of mice. The mechanism may be related to the increase in miR-124 expression, decrease in Hedgehog and HMGB1 signaling pathway activities, and reduction in inflammatory responses in the liver. Hedgehog is a regulatory target for miR-124.

Adaptive niche genetic algorithm (ANGA) and lung ultrasound were combined, the death warning mathematical model was established for patients with sepsis-lung injury, and the epidemiological characteristics were analyzed to explore the efficacy of Vancomycin in the treatment of sepsis-lung injury. First, 88 sepsis patients with lung injury were selected as the research objects. General clinical data and pulmonary ultrasound results were collected. On this basis, epidemiological analysis was carried out, and the death warning model of patients with sepsis-lung injury was established based on ANGA algorithm. Then, the total cure rate, Staphylococcus aureus (SA) clearance rate, methicillin-resistant SA (MRSA) clearance rate, and the incidence of adverse reactions after intravenous infusion of Vancomycin were analyzed. The results showed that the ANGA mathematical model combined with the random forest (RF) classifier proposed had better classification effect and robustness relative to the traditional principal component analysis and NGA. The early warning accuracy of the proposed ANGA + RF mathematical model was higher than 95% in contrast to that of the APACHE-II score and the SOFA score. Compared with patients in the severe group, the MRSA infection rate and the levels of procalcitonin (PCT), C-reactive protein (CRP), and activated partial thromboplastin time (APTT) of SA sepsis-lung injury patients were greatly reduced, while thrombin time (TT) and D-D dimer in the death group were considerably increased (p < 0.05), and the PLT level was greatly reduced (p < 0.05). In addition, the total cure rate, SA clearance rate, and MRSA clearance rate of Vancomycin-treated SA sepsis-lung injury patients were significantly increased (p < 0.05) compared with patients in the conventional treatment control group. However, the probability of adverse reactions was increased notably (p < 0.05). ANGA combined with RF classifier can improve the accuracy of death warning in patients with sepsis-lung injury. Vancomycin can effectively eliminate MRSA infection rate in patients with sepsis-lung injury and improve the treatment effect of patients.

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