Efficacy of Deep Venous Catheterization in the Management of Pneumothorax: A Clinical Study Utilizing Conventional Closed Thoracic Drainage.

Background: Currently, conventional closed thoracic drainage for pneumothorax involves a painful procedure with a higher risk and wider (1~1.5 cm) incision. Minimally invasive catheterized drainage techniques are urgently needed to address this challenge. Objective: This retrospective study aims to observe the effects of conventional closed thoracic drainage with deep venous catheterization drainage techniques on pneumothorax patients. Design: It was a retrospective study. Setting: This study was conducted at Huaian No.1 People's Hospital, Affiliated with Nanjing Medical University. Participants: A total of 105 pneumothorax patients who underwent conventional closed thoracic drainage (CCTD) or deep venous catheterization drainage technique (DVCDT) procedures at the hospital from 1st February 2020 to 30th October 2022 were selected. Interventions: Patients received either CCTD or DVCDT. Primary Outcome Measures: Included: (1) clinical variables; (2) catheterization procedure-related features; and (3) visual analogue scale (VAS) scores from pneumothorax patients. Results: Both conventional closed thoracic drainage and deep venous catheterization drainage techniques were successfully performed in all 105 (100%) patients, comprising 67 (63.8%) spontaneous pneumothorax, 20 (19%) iatrogenic pneumothorax, and 18 (17.1%) traumatic pneumothorax cases. Significant differences were observed between the enrolled spontaneous pneumothorax and traumatic pneumothorax patients in the two groups (CCTD and DVCDT) (P = .01 and P < .0001). Additionally, 55 (52.4%) patients underwent deep venous catheterization, while 50 (47.6%) patients underwent conventional closed thoracic drainage. The deep venous catheterization insertion procedure had a shorter mean timing (7.51±1.66 min) compared to the conventional closed thoracic drainage procedure (12.44±1.73 min) (P < .0001). Furthermore, VAS scores were significantly lower in pneumothorax patients undergoing deep venous catheterization (2.1±0.99) compared to conventional closed thoracic drainage (5.1±0.81) (P < .0001). Conclusion: Deep venous thoracic drainage technique appears to be safer and more beneficial than conventional closed thoracic drainage procedures for treating pneumothorax. This technique offers advantages such as minimal scarring, lower VAS scores, and shorter insertion time, thereby improving safety and surgical outcomes.

Fine-tuning rice heading date through multiplex editing of the regulatory regions of key genes by CRISPR-Cas9.

Heading date (or flowering time) is a key agronomic trait that affects seasonal and regional adaption of rice cultivars. An unoptimized heading date can either not achieve a high yield or has a high risk of encountering abiotic stresses. There is a strong demand on the mild to moderate adjusting the heading date in breeding practice. Genome editing is a promising method which allows more precise and faster changing the heading date of rice. However, direct knock out of major genes involved in regulating heading date will not always achieve a new germplasm with expected heading date. It is still challenging to quantitatively adjust the heading date of elite cultivars with best adaption for broader region. In this study, we used a CRISPR-Cas9 based genome editing strategy called high-efficiency multiplex promoter-targeting (HMP) to generate novel alleles at cis-regulatory regions of three major heading date genes: Hd1, Ghd7 and DTH8. We achieved a series of germplasm with quantitative variations of heading date by editing promoter regions and adjusting the expression levels of these genes. We performed field trials to screen for the best adapted lines for different regions. We successfully expanded an elite cultivar Ningjing8 (NJ8) to a higher latitude region by selecting a line with a mild early heading phenotype that escaped from cold stress and achieved high yield potential. Our study demonstrates that HMP is a powerful tool for quantitatively regulating rice heading date and expanding elite cultivars to broader regions.

Progranulin modulates the progression of non-small cell lung cancer through lncRNA H19.

OBJECTIVE: This study aimed to explore the specific mechanism of action of Progranulin (PGRN) in non-small cell lung cancer (NSCLC) and its interaction with lncRNA H19. METHODS: Normal and cancerous lung tissues were collected from patients with NSCLC and healthy volunteers. We assessed the expression of PGRN in both groups using immunohistochemistry, quantitative-reverse transcription-polymerase chain reaction (qRT-PCR), and western blotting (WB). RESULTS: Compared to the controls, PGRN expression was noticeably higher in tumor tissues. The high expression of PGRN in patients with NSCLC was inversely correlated to the prognosis and strongly associated with the biological features and clinicopathologic data. High PGRN expression significantly improved the ability of NSCLC cells to proliferate and migrate and was positively correlated with tumor formation, based on in vitro and in vivo cellular tests. Expression of lncRNA H19 was also found to be elevated in NSCLC tissue and cells. The expression of H19 was correlated with tumor growth in vivo and in vitro, and H19 regulated PGRN by mediating the expression of miR-29b-3p. CONCLUSIONS: H19 and PGRN can serve as biomarkers and therapeutic targets in NSCLC.

Phytophthora RxLR effector PcSnel4B promotes degradation of resistance protein AtRPS2.

Phytophthora capsici deploys effector proteins to manipulate host immunity and facilitate its colonization. However, the underlying mechanisms remain largely unclear. In this study, we demonstrated that a Sne-like (Snel) RxLR effector gene PcSnel4 is highly expressed at the early stages of P. capsici infection in Nicotiana benthamiana. Knocking out both alleles of PcSnel4 attenuated the virulence of P. capsici, while expression of PcSnel4 promoted its colonization in N. benthamiana. PcSnel4B could suppress the hypersensitive reaction (HR) induced by Avr3a-R3a and RESISTANCE TO PSEUDOMONAS SYRINGAE 2 (AtRPS2), but it did not suppress cell death elicited by Phytophthora infestin 1 (INF1) and Crinkler 4 (CRN4). COP9 signalosome 5 (CSN5) in N. benthamiana was identified as a host target of PcSnel4. Silencing NbCSN5 compromised the cell death induced by AtRPS2. PcSnel4B impaired the interaction and colocalization of Cullin1 (CUL1) and CSN5 in vivo. Expression of AtCUL1 promoted the degradation of AtRPS2 and disrupted HR, while AtCSN5a stabilized AtRPS2 and promoted HR, regardless of the expression of AtCUL1. PcSnel4 counteracted the effect of AtCSN5 and enhanced the degradation of AtRPS2, resulting in HR suppression. This study deciphered the underlying mechanism of PcSnel4-mediated suppression of HR induced by AtRPS2.

Construction of a novel ESIPT and AIE-based fluorescent sensor for sequentially detecting Cu2+ and H2S in both living cells and zebrafish.

The development of effective methods for tracking Cu2+ and H2S in living organisms is urgently required due to their vital function in a variety of pathophysiological processes. In this work, a new fluorescent sensor BDF with excited-state intramolecular proton transfer (ESIPT) and aggregation-induced emission (AIE) features for the successive detection of Cu2+ and H2S was constructed by introducing 3,5-bis(trifluoromethyl)phenylacetonitrile into the benzothiazole skeleton. BDF showed a fast, selective and sensitive fluorescence "turn off" response to Cu2+ in physiological media, and the situ-formed complex can serve as a fluorescence "turn on" sensor for highly selective detection of H2S through the Cu2+ displacement approach. In addition, the detection limits of BDF for Cu2+ and H2S were determined to be 0.05 and 1.95 µM, respectively. Encouraged by its favourable features, including strong red fluorescence from the AIE effect, large Stokes shift (285 nm), high anti-interference ability and good function at physiological pH as well as a low toxicity, BDF was successfully applied for the consequent imaging of Cu2+ and H2S in both living cells and zebrafish, making it an ideal candidate for detecting and imaging of Cu2+ and H2S in live systems.

A single region of the Phytophthora infestans avirulence effector Avr3b functions in both cell death induction and plant immunity suppression.

As a destructive plant pathogen, Phytophthora infestans secretes diverse host-entering RxLR effectors to facilitate infection. One critical RxLR effector, PiAvr3b, not only induces effector-triggered immunity (ETI), which is associated with the potato resistance protein StR3b, but also suppresses pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI). To date, the molecular basis underlying such dual activities remains unknown. Based on phylogenetic analysis of global P. infestans isolates, we found two PiAvr3b isoforms that differ by three amino acids. Despite this sequence variation, the two isoforms retain the same properties in activating the StR3b-mediated hypersensitive response (HR) and inhibiting necrosis induced by three PAMPs (PiNpp, PiINF1, and PsXeg1) and an RxLR effector (Pi10232). Using a combined mutagenesis approach, we found that the dual activities of PiAvr3b were tightly linked and determined by 88 amino acids at the C-terminus. We further determined that either the W60 or the E134 residue of PiAvr3b was essential for triggering StR3b-associated HR and inhibiting PiNpp- and Pi10232-associated necrosis, while the S99 residue partially contributed to PTI suppression. Additionally, nuclear localization of PiAvr3b was required to stimulate HR and suppress PTI, but not to inhibit Pi10232-associated cell death. Our study revealed that PiAvr3b suppresses the plant immune response at different subcellular locations and provides an example in which a single amino acid of an RxLR effector links ETI induction and cell death suppression.

A red-emissive benzothiazole-based luminophore with ESIPT and AIE natures and its application for detecting and imaging hypochlorous acid.

A new "ESIPT + AIE" based dye of benzothiazole with red emission and a large Stokes shift was constructed by combining 2-(2'-hydroxyphenyl)benzothiazole as the ESIPT unit and α-cyanostilbene as the AIE unit. The compound BACN was found to be a ideal HClO chemosensor, and presented palpable fluorescence and colorimetric responses toward HClO via the HClO-trigged oxidation cleavage of the ethylene bridge activated by the electron withdrawing cyano group. BACN was capable of recognizing HClO rapidly (12 s) and sensitively under physiological conditions, with good selectivity over other biologically pertinent substances. Thanks to strong red emission (λem = 606 nm) and large Stokes shift (213 nm) resulted from the combination of ESIPT and AIE effects, it was successfully utilized for the recognition of exogenous and endogenous HClO in living HeLa cells.

CRIF1 promotes the progression of non-small-cell lung cancer by SIRT3- mediated deacetylation of PYCR1.

Lung cancer is the cancer with the highest mortality in the world. So further exploration of the pathogenesis of lung cancer is of great significance. In this study, the specific role and related mechanism of CRIF1 in non-small cell lung cancer (NSCLC) were explored in this research. TheRT-PCR, western blot and IHC assays were used to examine the expression level of CRIF1 in NSCLC tissue, tissue adjacent to carcinoma, NSCLC cell lines and human normal lung epithelial cells. Next, colony formation assay, Alamar blue Kit and EdU assays were employed to examine the proliferation of transfected A549 and NCI-H2009 cells. Measurement of mitochondrial permeability transition pore opening, ATP production and cellular oxygen consumption were used to evaluate the mitochondrial apoptosis of transfected NSCLC cells. Enzymatic activity assays for PYCR1, western blot and flow cytometry assays were used to explore the relationship between PYCR1 and CRIF1. The subcutaneous xenograft tumor mice model was established to explore the role of CRIF1 in vivo. Collectively, results revealed that CRIF1 was upregulated in NSCLC cells and tissues (p < 0.001). CRIF1 promoted proliferation of NSCLC cells (p < 0.001). CRIF1 inhibited mitochondrial apoptosis in NSCLC cells (p < 0.05). Moreover, CRIF1 promoted PYCR1 deacetylation and increased its activity through SIRT3 (p < 0.05). Deacetylation of PYCR1 reversed the antitumor effect of CRIF1 knockdown (p < 0.05). Finally, knockdown of CRIF1 inhibited the tumor growth of NSCLC in vivo (p < 0.05).This research found that CRIF1 promoted the progression of non-small-cell lung cancer by SIRT3- mediated deacetylation of PYCR1.

An AIE based fluorescent chemosensor for ratiometric detection of hypochlorous acid and its application.

Detecting and imaging intracellular hypochlorous acid (HClO) is of great importance owning to its prominent role in numerous pathological and physiological processes. In this contribution, a novel AIE-based fluorescent chemosensor has been developed by employing a benzothiazole derivative. The synthesized probe displayed remarkable ratiometric fluorescent response to HClO with a large emission shift (139 nm), resulting in naked-eye fluorescence changes from red to blue. Under the optimal conditions, this probe was capable of quantitatively detecting HClO within 10 s, and possessed good sensitivity and high selectivity toward HClO over other biologically relevant species. Moreover, it has been successfully utilized to image the exogenous and endogenous HClO in living cells through dual channels, and conveniently detect hypochlorous acid solution on test strips with better accuracy, demonstrating its potential for monitoring HClO in biological and environment fields.

Analysis of resistance risk and resistance-related point mutations in Cyt b of QioI fungicide ametoctradin in Phytophthora litchii.

BACKGROUND: Litchi downy blight, caused by Phytophthora litchii, is one of the most important diseases of litchi. Ametoctradin, as the only QioI (quinone inside and outside inhibitor) fungicide, has been registered in China in 2019. However, the ametoctradin-resistance risk and molecular basis in Phytophthora litchii have not been reported. RESULTS: In this study, the sensitivity profile of 144 Phytophthora litchii strains to ametoctradin was determined, with a mean median effective concentration (EC50 ) value of 0.1706 ± 0.091 µg mL-1 . Nine stable resistant Phytophthora litchii mutants [resistance factor (RF) > 400] were derived from sensitive isolates using fungicide adaption. The compound fitness index of three resistant-mutants (HN10-1-1, HN10-1-2 and HN10-2-1) was similar or higher than that of their parental isolates in vitro. All these ametoctradin-resistant mutants were sensitive to metalaxyl, dimethomorph, oxathiapiprolin and cyazofamid. Two point mutations, leading to the S33L and D228N changes in PlCyt b (cytochrome b) were found in ametoctradin-resistant mutants. Eight ametoctradin-resistant mutants containing S33L showed increased sensitivity to azoxystrobin and amisulbrom, and one mutant containing D228N exhibited increased sensitivity to cyazofamid. In vitro enzyme activity test showed that ametoctradin could not inhibit the activity of cytochrome bc1 complex with S33L and D228N point mutation. AS-PCR primers were designed based on the S33L change to detect the ametoctradin-resistant strains in the future. CONCLUSION: These results suggest that Phytophthora litchii has a medium to high resistance risk to ametoctradin in the laboratory. Two changes, S33L and D228N, in PlCyt b are likely to be associated with the observed ametoctradin resistance. © 2022 Society of Chemical Industry.

Fluorescent porous organic cage with good water solubility for ratiometric sensing of gold(III) ion in aqueous solution.

Fluorescent porous organic cage with good water solubility is of great interest but still challenging for its fluorescent sensing application. Poor water solubility and single signal of most previous probes are unfavorable for the monitoring of Au3+ generated from the potential dissociation of gold nanoparticles in environmental and biological samples. Here we report a water-soluble porous organic cage as a ratiometric fluorescent probe for Au3+ in aqueous solution. The prepared porous organic cage with good water solubility showed specific redox interaction with Au3+ in pH 5, leading to the change of dual emission at 420 and 484 nm. Based on the change of fluorescence ratio, a simple ratiometric sensing method for Au3+ from the dissociation of gold nanoparticles in aqueous solution was developed. The proposed method gave a calibration function of F484/F420 = 0.0370[Au3+] + 0.5689 (where F484/F420 is the intensity ratio of fluorescence at 484 nm to that at 420 nm; [Au3+] in µM) (R2 = 0.9975) in the concentration range of 1-60 µM, the limit of detection (3s) of 8 nM, and the relative standard deviation of 0.26% for 10 replicate detections of 50 µM Au3+. The recoveries of spiked Au3+ in domestic wastewater and human serum samples ranged from 94.66% to 105.61%.

An N-nitrosation reaction-based fluorescent probe for detecting nitric oxide in living cells and inflammatory zebrafish.

Nitric oxide (NO), an essential biological messenger molecule, participates in various physiological and pathological processes. The sensitive and specific detection of NO is of great significance for understanding the biological function of NO. Here, we synthesized a fluorescent probe (Rho-NO) for highly selective detection of NO both in vitro and in vivo. The high selectivity of Rho-NO is attributed to the fact that NO is easily replaced by electron donor amino group to form N-nitrosation products, causing rhodamine spiro ring open and fluorescence emit. Rho-NO showed a good linear response to NO (0-100 µM) with a low detection limit (0.06 µM). Importantly, it exhibited excellent specificity for NO detection in human serum and was also applied for imaging NO in living cells and inflammatory model of zebrafish. This work proves the potential of Rho-NO in pathological research and disease diagnosis.

Characterization of Prochloraz Resistance in Fusarium fujikuroi from Heilongjiang Province in China.

Prochloraz is widely used to control rice bakanae disease caused by Fusarium fujikuroi. The current study was aimed at monitoring the development of F. fujikuroi resistance to prochloraz in the Heilongjiang Province and analyzing the fitness of F. fujikuroi strains with different resistance levels. The results indicated that most of the 89 F. fujikuroi strains collected from the Heilongjiang Province were resistant to prochloraz, with resistance frequency reaching 92.1%. To assess the field resistance risk of prochloraz, 21 F. fujikuroi strains with different resistance levels were selected to investigate their biological characteristics and assess their fitness. Mycelial growth, sporulation, and germination rates were significantly different among the tested strains. However, when grouped into two subpopulations, no significant difference was tested between prochloraz-resistant and prochloraz-sensitive strains. Pathogenicity assays revealed that the disease severity index of prochloraz-resistant strains was higher than that of prochloraz-sensitive strains. Cross-resistance assays showed no cross-resistance between prochloraz and five other fungicides, namely phenamacril, ipconazole, tebuconazole, carbendazim, and fluopyram. Ffcyp51A gene overexpression was observed in the prochloraz-resistant F. fujikuroi strains after exposure to prochloraz. Collectively, these results indicated that F. fujikuroi resistance against prochloraz was severe. Furthermore, prochloraz-resistant strains were highly fit and could potentially become a dominant population in rice fields, consequently resulting in yield loss.

Simultaneous visualization and quantification of copper (II) ions in Alzheimer's disease by a near-infrared fluorescence probe.

The abnormal accumulation of copper ions (Cu2+) is considered to be one of the pathological factors of Alzheimer's disease (AD), but the internal relationship between Cu2+ and AD progression is still not fully clear. In this work, a sensitive and selective near-infrared fluorescent copper ion probe (DDP-Cu) was designed for quantification and visualization of Cu2+ level in lysates, living cells, living zebrafish and brain tissues of drosophila and mice with AD. By using this probe, we demonstrated that the content of Cu2+ in the brains of AD mice and drosophila enhanced nearly 3.5-fold and 4-fold than that of normal mice and drosophila, respectively. More importantly, pathogenesis analysis revealed that elevated Cu2+ led to changes in factors closely associated with AD, such as the increasing of reactive oxygen species(ROS), the aggregation of amyloid-ß protein (Aß) and nerve cell cytotoxicity. These findings could promote the understanding of the roles between Cu2+ and AD.

Resistance to pydiflumetofen in Botrytis cinerea: risk assessment and detection of point mutations in sdh genes that confer resistance.

BACKGROUND: Gray mold caused by Botrytis cinerea Pers. is one of the most significant airborne diseases. It can infest a wide range of crops, causing significant losses in yield and quality worldwide. Pydiflumetofen, a new generation succinate dehydrogenase inhibitor (SDHI), is currently being registered in China to control gray mold in a variety of crops. The baseline sensitivity, resistance risk, and resistance mechanism of Botrytis cinerea to pydiflumetofen were assessed in this study. RESULTS: A total of 138 strains of B. cinerea from 10 different regions were tested for their sensitivity to pydiflumetofen, and the mean EC50 value was 0.0056 µg mL-1 . Eight mutants were obtained by fungicide adaption from five sensitive parental isolates, and the resistance factor (RF) ranged from 51 to 135. The mutants exhibited strong adaptive traits in conidial production, conidial germination, and pathogenicity. Positive cross-resistance was only observed between other SDHIs (i.e. boscalid, fluopyram, and isopyrazam). Two different types of pydiflumetofen-resistant mutants were identified: point mutation P225L in sdhB and double mutation G85A and I93V in sdhC. The in vivo control efficacy of pydiflumetofen on the resistant mutants carrying P225L in sdhB as well as G85A and I93V in sdhC was significantly decreased to 52.62% and 32.27%, respectively. CONCLUSION: The fitness was significantly higher for all pydiflumetofen-resistant mutants than the corresponding parental. Two types of point mutations, sdhB-P225L and sdhC-G85A and I93V, might confer resistance to pydiflumetofen in B. cinerea. A precautionary resistance management strategy should be implemented. © 2021 Society of Chemical Industry.

ELK1­mediated upregulation of lncRNA LBX2­AS1 facilitates cell proliferation and invasion via regulating miR­491­5p/S100A11 axis in colorectal cancer.

The aim of the present study was to investigate the role and regulatory mechanism of LBX2 antisense RNA 1 (LBX2­AS1) in colorectal cancer. Firstly, LBX2­AS1 expression was detected using reverse transcription­quantitative PCR in colorectal cancer tissues and cells, and its prognostic and diagnostic efficacy was assessed in a colorectal cancer cohort (n=145). Subcellular fractionation assay of LBX2­AS1 was performed. Secondly, the effects of LBX2­AS1 and microRNA (miR)­491­5p on colorectal cancer cell proliferation, apoptosis, migration and invasion were investigated by a series of functional assays. Thirdly, RNA immunoprecipitation, dual­luciferase reporter and gain and loss of function assays were carried out to analyze the interactions between ETS transcription factor ELK1 (ELK1) and LBX2­AS1, as well as LBX2­AS1, miR­491­5p and S100A11. The results showed that LBX2­AS1 was upregulated both in colorectal cancer tissues and cells, which was distributed in the cytoplasm and nucleus of colorectal cancer cells. Clinically, high LBX2­AS1 expression could be an independent prognostic factor for colorectal cancer. Furthermore, relative operating characteristic curve analysis showed that LBX2­AS1 was a sensitive diagnostic marker for colorectal cancer. Highly expressed ELK1, as a transcription factor, could bind to the two conserved sites in the promoter region of LBX2­AS1, thereby activating the transcription of LBX2­AS1. Silencing LBX2­AS1 markedly inhibited proliferative, migratory and invasive abilities of colorectal cancer cells. miR­491­5p expression was downregulated, while S100A11 expression was upregulated in colorectal cancer tissues and cells. Dual­luciferase reporter assays confirmed that LBX2­AS1 could block S100A11 degradation via competitively binding to miR­491­5p. Furthermore, LBX2­AS1 overexpression could notably reverse the inhibitory effect of miR­491­5p on proliferation and invasion of colorectal cancer cells. Taken together, LBX2­AS1 induced by transcription factor ELK1 may facilitate colorectal cancer cell proliferation and invasion via regulation of the miR­491­5p/S100A11 axis. Thus, LBX2­AS1 could be an underlying prognostic and diagnostic marker for colorectal cancer.

A morpholino hydrazone-based lysosome-targeting fluorescent probe with fast response and high sensitivity for imaging peroxynitrite in living cells.

Peroxynitrite (ONOO-) plays important roles in many pathophysiological processes and its subcellular detection draws increasing attention. In this study, we designed and prepared a novel lysosome-targetable fluorescent probe (E)-2-(benzo[d]thiazol-2- yl)-4-methyl-6-((morpholinoimino)methyl)phenol (BMP) for selective detection of ONOO- in living systems by incorporating a reactive morpholino hydrazone as new ONOO- response site into a benzothiazole derivative as fluorophore. After reaction with ONOO-, an obvious fluorescence increase (83-fold) was observed accompanied with distinct dual colorimetric and fluorescence changes. Probe BMP displayed the merits of fast response (<3 s), ultrasensitivity (LOD = 6 nM) and high selectivity towards ONOO- over other physiological species including ROS/RNS. Most importantly, the probe was capable of imaging ONOO- in lysosomes of living cells with good cell permeation and negligible cytotoxicity. Therefore, this research provides an effective tool to study the functions of ONOO- in lysosomes.

Transcriptional Variability Associated With CRISPR-Mediated Gene Replacements at the Phytophthora sojae Avr1b-1 Locus.

Transcriptional plasticity enables oomycetes to rapidly adapt to environmental challenges including emerging host resistance. For example, the soybean pathogen Phytophthora sojae can overcome resistance conferred by the host resistance gene Rps1b through natural silencing of its corresponding effector gene, Avr1b-1. With the Phytophthora CRISPR/Cas9 genome editing system, it is possible to generate site-specific knock-out (KO) and knock-in (KI) mutants and to investigate the biological functions of target genes. In this study, the Avr1b-1 gene was deleted from the P. sojae genome using a homology-directed recombination strategy that replaced Avr1b-1 with a gene encoding the fluorescent protein mCherry. As expected, all selected KO transformants gained virulence on Rps1b plants, while infection of plants lacking Rps1b was not compromised. When a sgRNA-resistant version of Avr1b-1 was reintroduced into the Avr1b-1 locus of an Avr1b KO transformant, KI transformants with a well-transcribed Avr1b-1 gene were unable to infect Rps1b-containing soybeans. However, loss of expression of the incoming Avr1b-1 gene was frequently observed in KI transformants, which resulted in these transformants readily infecting Rps1b soybeans. A similar variability in the expression levels of the incoming gene was observed with AVI- or mCherry-tagged Avr1b-1 constructs. Our results suggest that Avr1b-1 may be unusually susceptible to transcriptional variation.

MicroRNA-564 inhibits the progression of non-small cell lung cancer via targeting plexin A4.

Νon-small cell lung cancer (NSCLC) is the most frequently diagnosed type of cancer, and the most prevalent cause of cancer-associated mortality. The present study aimed to investigate whether microRNA (miR)-564 influences NSCLC progression by regulating NSCLC cell growth and migration, via targeting plexin A4. Therefore, the expression levels of miR-564 and plexin A4 were evaluated in NSCLC specimens or cells using reverse transcription-quantitative PCR. Furthermore, colony formation and Cell Counting Kit-8 assays were performed to determine the proliferative ability of NSCLC cells. The cell migration capacity was assessed using a Transwell assay. In addition, to examine the binding ability of miR-564 on the plexin A4 3'-untranslated region (3'UTR), a dual-luciferase reporter assay was performed. A mouse xenograft model was established to evaluate the effect of miR-564 knockdown on tumor growth in vivo, whereas the expression of plexin A4 and Ki67 in NSCLC tissues was detected using immunohistochemistry. Notably, miR-564 was downregulated in both NSCLC cell lines and tissues, while its overexpression, following transfection with miR-564 mimics, attenuated the proliferation and proliferation, migration and invasion of NSCLC cells. By contrast, silencing of miR-564 using a miR-564 inhibitor promoted NSCLC cell proliferation, migration and invasion. The luciferase assay revealed that miR-564 directly targeted the plexin A4 3'UTR in A549 and H460 cells. Additionally, the overexpression of plexin A4 rescued the effect of miR-564 on NSCLC cell proliferation, migration and invasion abilities. Further in vivo studies demonstrated that miR-564 knockdown promoted NSCLC growth, while miR-564 overexpression resulted in the opposite effect in nude mice. Overall, the results of the present study revealed that miR-564 promotes the proliferation and migration of NSCLC cells, both in vitro and in vivo, via targeting plexin A4. Therefore, miR-564 may be considered as a possible therapeutic target for NSCLC.

A colorimetric and far-red fluorescent probe for rapid detection of bisulfite/sulfite in full water-soluble based on biquinolinium and its applications.

Bisulfite (HSO3-) and sulfite (SO32-) are involved in numerous physiological processes of living systems. However, high levels of these substances are often correlated to many diseases. Herein, we designed and synthesized a simple full water-soluble colorimetric and far-red fluorescent probe (E)-1-methyl-4-(2-(1-methylquinolin-1-ium-3-yl)vinyl)quinolin-1-ium iodide trifluoromethanesulfonate (DQ) for HSO3-/SO32- detection by coupling 1,4-dimethylquinolinium with 3-quinolinium carboxaldehyde for the first time. The probe DQ showed high selectivity for HSO3- detection via a 1,4-nucleophilic addition reaction with distinct color changes from colorless to purple-red and remarkable far-red fluorescence enhancement in pure aqueous solutions. Specifically, the probe displayed a fast response (<15 s) for bisulfite, which renders it suitable for real time detection of HSO3-. Under the optimized conditions, the far-red fluorescence intensity was linear to the concentrations of HSO3- in the range from 0 to 25 µM and the detection limit was as low as 0.11 µM. Additionally, the probe could be applied to sense HSO3- on paper strips, real sample including vermicelli and sugar and image HSO3- in living cells, which indicated that probe DQ has potential application in food samples and living systems.