Hyperechoic Spots in the Renal Medulla as a Potential Indicator of Early Gouty Nephropathy.

INTRODUCTION: The aim of the study was to explore the causes and clinical significance of hyperechoic renal medulla observed by ultrasonography in patients with primary gout. METHODS: This study included 2,107 patients with primary gout treated in the Gout Clinic of our hospital from 2016 to 2022. The clinical data and biochemical data of these patients were collected and analyzed. According to the presence or absence of punctate hyperechogenicity in the renal medulla on ultrasound examination, the patients were divided into the hyperechoic medulla (HM) and the normal hypoechoic medulla (NM) groups, and the HM group was further divided into the partial HM (P-HM) and fulfilled HM (F-HM) subgroups according to the distribution range of hyperechogenicity. RESULTS: Among the 2,107 patients with primary gout, 380 had hyperechoic renal medulla on renal ultrasound, including 106 patients with F-HM and 274 with P-HM. There were significant differences in the gout duration, urate arthropathy number, serum urate (SU) level, clinical tophi number, blood urea nitrogen, serum creatinine (sCr), and estimated glomerular filtration rate between the HM and NM groups or between the F-HM and P-HM subgroups (p < 0.05). Multivariate regression analysis showed that the presence of HM was positively correlated with gout duration, urate arthropathy number, gout attack frequency, SU, and sCr. The number of clinical tophi and sCr were closely related to F-HM. CONCLUSION: Ultrasound examination showed that a high medulla echo in patients with gout was often related to renal function damage. P-HM may be a transitory condition between NM and F-HM in patients with gout.

CsLOB1 regulates susceptibility to citrus canker through promoting cell proliferation in citrus.

Citrus sinensis lateral organ boundary 1 (CsLOB1) was previously identified as a critical disease susceptibility gene for citrus bacterial canker, which is caused by Xanthomonas citri subsp. citri (Xcc). However, the molecular mechanisms of CsLOB1 in citrus response to Xcc are still elusive. Here, we constructed transgenic plants overexpressing and RNAi-silencing of CsLOB1 using the canker-disease susceptible 'wanjincheng' orange (C. sinensis Osbeck) as explants. CsLOB1-overexpressing plants exhibited dwarf phenotypes with smaller and thicker leaf, increased branches and adventitious buds clustered on stems. These phenotypes were followed by a process of pustule- and canker-like development that exhibited enhanced cell proliferation. Pectin depolymerization and expansin accumulation were enhanced by CsLOB1 overexpression, while cellulose and hemicellulose synthesis were increased by CsLOB1 silence. Whilst overexpression of CsLOB1 increased susceptibility, RNAi-silencing of CsLOB1 enhanced resistance to canker disease without impairing pathogen entry. Transcriptome analysis revealed that CsLOB1 positively regulated cell wall degradation and modification processes, cytokinin metabolism, and cell division. Additionally, 565 CsLOB1-targeted genes were identified in chromatin immunoprecipitation-sequencing (ChIP-seq) experiments. Motif discovery analysis revealed that the most highly overrepresented binding sites had a conserved 6-bp 'GCGGCG' consensus DNA motif. RNA-seq and ChIP-seq data suggested that CsLOB1 directly activates the expression of four genes involved in cell wall remodeling, and three genes that participate in cytokinin and brassinosteroid hormone pathways. Our findings indicate that CsLOB1 promotes cell proliferation by mechanisms depending on cell wall remodeling and phytohormone signaling, which may be critical to citrus canker development and bacterial growth in citrus.

Overexpression of Salicylic Acid Carboxyl Methyltransferase (CsSAMT1) Enhances Tolerance to Huanglongbing Disease in Wanjincheng Orange (Citrus sinensis (L.) Osbeck).

Citrus Huanglongbing (HLB) disease or citrus greening is caused by Candidatus Liberibacter asiaticus (Las) and is the most devastating disease in the global citrus industry. Salicylic acid (SA) plays a central role in regulating plant defenses against pathogenic attack. SA methyltransferase (SAMT) modulates SA homeostasis by converting SA to methyl salicylate (MeSA). Here, we report on the functions of the citrus SAMT (CsSAMT1) gene from HLB-susceptible Wanjincheng orange (Citrus sinensis (L.) Osbeck) in plant defenses against Las infection. The CsSAMT1 cDNA was expressed in yeast. Using in vitro enzyme assays, yeast expressing CsSAMT1 was confirmed to specifically catalyze the formation of MeSA using SA as a substrate. Transgenic Wanjincheng orange plants overexpressing CsSAMT1 had significantly increased levels of SA and MeSA compared to wild-type controls. HLB resistance was evaluated for two years and showed that transgenic plants displayed significantly alleviated symptoms including a lack of chlorosis, low bacterial counts, reduced hyperplasia of the phloem cells, and lower levels of starch and callose compared to wild-type plants. These data confirmed that CsSAMT1 overexpression confers an enhanced tolerance to Las in citrus fruits. RNA-seq analysis revealed that CsSAMT1 overexpression significantly upregulated the citrus defense response by enhancing the transcription of disease resistance genes. This study provides insight for improving host resistance to HLB by manipulation of SA signaling in citrus fruits.

Preparation of bio-absorbents by modifying licorice residue via chemical methods and removal of copper ions from wastewater.

In this paper, a series of bio-adsorbents (LR-NaOH, LR-Na2CO3 and LR-CA) were successfully prepared by modifying Licorice Residue (LR) with NaOH, Na2CO3 and citric acid, and were used as the adsorbents to remove Cu2+ from wastewater. The morphology and structure of bio-adsorbents were characterized by Fourier Transform Infrared, scanning electron microscopy (SEM), thermogravimetry (TG) and X-ray diffraction (XRD). Using static adsorption experiments, the effects of the adsorbent dosage, the solution pH, the adsorption time, and the initial Cu2+ concentration on the adsorption performance of the adsorbents were investigated. The results showed that the adsorption process of Cu2+ by the bio-adsorbents can be described by pseudo-second-order kinetic model and the Langmuir model. The surface structure of the LR-NaOH, LR-Na2CO3 and LR-CA changed obviously, and the surface-active groups increased. The adsorption capacity of raw LR was 21.56 mg/g, while LR-NaOH and LR-Na2CO3 significantly enhanced this value up to 43.65 mg/g and 43.55 mg/g, respectively. After four adsorption-desorption processes, the adsorption capacity of LR-NaOH also maintained about 73%. Therefore, LR-NaOH would be a promising adsorbent for removing Cu2+ from wastewater, and the simple strategy towards preparation of adsorbent from the waste residue can be a potential approach for use in the water treatment.

Clinical aPDT's effect on Candida albicans: Antifungal susceptibility, virulence gene expression, and correlation with leukocyte and neutrophil counts.

BACKGROUND: Our previous clinical trial demonstrated that antimicrobial photodynamic therapy (aPDT) with methylene blue (MB) and potassium iodide (KI) effectively killed Candida albicans (C. albicans) in adult AIDS patients with oral candidiasis, regardless of biofilm formation or 25S rDNA genotype. This study evaluated changes in antifungal susceptibility and virulence gene expression in C. albicans before and after aPDT, and explored factors related to clinical aPDT efficacy. METHODS: Twenty-one adult AIDS patients with C. albicans oral candidiasis were divided into Group a (400 µM MB, N = 11) and Group b (600 µM MB, N = 10). Both groups received two aPDT treatments, where MB was applied for 5 min, followed by 300 mM KI, and illuminated for 30 min (37.29 J/cm²). C. albicans isolates were collected before and after treatment to assess antifungal susceptibility (fluconazole, itraconazole, flucytosine, amphotericin B) and gene expression (CAT1, HWP1). Peripheral blood tests were analyzed for correlations with aPDT efficacy. RESULTS: aPDT reduced minimum inhibitory concentration (MIC) values for amphotericin B, fluconazole, and flucytosine, with significant reductions primarily after the first treatment. MIC reductions differed between groups, with Group a showing greater decreases in flucytosine and fluconazole MICs, and Group b in amphotericin B MICs. No significant changes in CAT1 or HWP1 expression were observed. Clinical efficacy of aPDT negatively correlated with leukocyte and neutrophil levels. CONCLUSIONS: aPDT effectively reduces MICs of antifungal drugs against C. albicans isolated from treated patients, particularly after the first treatment. The concentration of MB required to reduce MICs varies among different antifungal drugs. aPDT does not alter CAT1 or HWP1 expression, and its clinical efficacy in eradicating C. albicans is negatively associated with leukocyte and neutrophil levels.

Primary Malignant Melanoma in the Parotid Gland: A Case Report and Literature Review.

Primary melanoma of the parotid gland is an extremely rare and challenging tumor with a poor prognosis, and its ultrasonic characteristics have yet to be reported. This article presents a case of a 77-year-old man with a left parotid mass that was confirmed as a melanoma following surgery. The ultrasonic features of melanoma were examined in detail, with a particular focus on their diagnostic value. Furthermore, we summarized the clinical characteristics, treatment options, and outcomes associated with primary melanoma of the parotid gland based on a thorough analysis of the available literature.

Preoperative Clinical and Ultrasonographic Predictors for Aggressive Behaviors in Sporadic Medullary Thyroid Carcinomas.

Objective: To explore the clinical and ultrasonographic predictors for aggressive behaviors preoperatively in sporadic medullary thyroid carcinomas (MTCs). Materials and Methods: The preoperative clinical and ultrasonographic characteristics of patients diagnosed with MTCs between January 2009 and May 2022 were retrospectively reviewed. MTCs were described and categorized according to the American College of Radiology (ACR) thyroid imaging reporting and data system classification by 2 radiologists. Interobserver agreement was evaluated by kappa test. Univariate and multivariate analyses were performed to identify predictors of aggressive behaviors in MTCs. The log-rank test was utilized to compare differences in Kaplan-Meier (K-M) curves for postoperative disease-free survival (PDFS). Results: A total of 120 patients were enrolled in the final study. Male sex was significant risk factor for metastasis, perithyroidal invasion, and lateral cervical lymph node (LCLN) metastasis [odds ratio (OR): 3.109, P = .019; OR: 5.316, P = .018; OR: 5.154 P = .012, respectively]. The kappa values for all ultrasonic characteristics were high (ranged from 0.811 to 0.941). Size, focality, and margin of the nodule were independent risk factors for metastasis, as well as for LCLN metastasis. Whereas margin (P < .001) and a subcapsular location (P = .021) were risk factors for perithyroidal invasion. According to K-M analysis, PDFS of patients differed significantly between groups with/without metastasis (P < .001), groups with/without perithyroidal extension (P < .001) and groups with/without LCLN metastasis (P < .001). Conclusions: Male sex is an independent risk factor for metastasis, perithyroidal invasion, and LCLN metastasis. The large size (≥2.55 cm for metastasis, ≥2.15 cm for LCLN metastasis, respectively), multifocality, and irregular margin of nodules were independent risk factors for both metastasis and LCLN metastasis. Extrathyroidal extension and a subcapsular location were risk factors for perithyroidal invasion. Moreover, patients with metastasis/perithyroidal extension/LCLN metastasis exhibited worse PDFS.

The usefulness of ultrasound in identifying the underlying findings linked to pain in podagra patients.

AIM: To explore whether ultrasound (US) can be employed to identify the underlying characteristics associated with pain in patients with podagra by evaluating the relationship between ultrasound findings and clinical pain. MATERIAL AND METHODS:  Patients with podagra were recruited and grouped into a pain group (G1, 82 patients) and a non pain group (G2, 123 patients). US features were collected and compared. US data were analyzed by binary logistic regression analysis and ROC analysis. Interobserver reliability was assessed, too. RESULTS: A total of 205 patients (196 male and 9 female) were enrolled in this study. In multivariate analysis, the thickness of the synovium (OR=1.928, CI=1.074-3.463), CD (color Doppler) signal of the synovium (OR=1.458, CI=1.011-2.103), and CD signal of the tophi (OR=1.576, CI=1.142-2.177) were identified as risk factors for clinical pain. Areas under the ROC curves (AUC) were 0.713, 0.686 and 0.641 for the three indicators, respectively. The best cutoff points were 1 mm for the thickness of the synovium, grade 1 for the CD signal of the synovium and grade 2 for the CD signal of the tophi. CONCLUSIONS: Ultrasound can provide valuable information for determining underlying features associated with pain in patients with podagra.

[The role of jasmonic acid in stress resistance of plants: a review].

Jasmonic acid (JA), a plant endogenously synthesized lipid hormone, plays an important role in response to stress. This manuscript summarized the biosynthesis and metabolism of JA and its related regulatory mechanisms, as well as the signal transduction of JA. The mechanism and regulatory network of JA in plant response to biotic and abiotic stresses were systematically reviewed, with the latest advances highlighted. In addition, this review summarized the signal crosstalk between JA and other hormones in regulating plant resistance to various stresses. Finally, the problems to be solved in the study of plant stress resistance mediated by JA were discussed, and the application of new molecular biological technologies in regulating JA signaling to enhance crop resistance was prospected, with the aim to facilitate future research and application of plant stress resistance.

Concordance of Ultrasound and Dual-Energy CT in Diagnosing Gouty Arthritis in the Knee Joint: A Retrospective Observational Study.

RATIONALE AND OBJECTIVES: To assess the consistency between ultrasound and dual-energy computed tomography (DECT) for the diagnosis of gout in the knee joint. MATERIALS AND METHODS: The ultrasound and DECT images of 176 knee joints from 167 patients diagnosed with gout at the Gout Specialty Clinic of Qingdao University Affiliated Hospital from February 2022 to December 2023 were retrospectively analyzed. The knee joint was segmented into five anatomical regions: intra-articular, anterior, posterior, medial, and lateral. The location of monosodium urate (MSU) crystal deposition was recorded. Tophi were classified as hypoechogenic, isoechogenic, hyperechogenic, or strongly echogenic. The Kappa test was used to assess the consistency between the two examination methods in different regions of the knee joint. The McNemar chi-square test was utilized to conduct a differential analysis between the DECT and ultrasound results. The chi-square test was used to assess differences in the rate of tophi detection with different echogenicities by DECT. Pearson's correlation coefficient was used to assess the correlation between MSU crystal deposition volume and clinically relevant indicators. RESULTS: Double contour (61.4%) was the most common intra-articular ultrasound sign. In the extra-articular region, MSU crystals were commonly deposited in and around the popliteal groove region (ultrasound: 52.3%; DECT: 60.0%). Corresponding MSU deposits on DECT were found in 7 of 54 joints with aggregates detected on ultrasound, and in 15 of 108 joints with DC. Tophi with hyperechogenicity or strong echogenicity were more likely to be detected on DECT than those with hypoechoic or isoechoic features (84.3% and 90.9% vs. 55.1% and 27.8%, respectively). For the assessment of MSU deposits, ultrasound showed an overall higher positive rate than DECT (81.1% vs. 72.2%), with poor consistency between the two examinations (κ = 0.177). In distinct anatomical regions, ultrasound and DECT showed high consistency in the medial (κ = 0.651) and lateral (κ = 0.705) views, with no significant difference. The intra-articular (κ = 0.316) and anterior (κ = 0.346) regions exhibited only fair consistency, with statistically significant diagnostic differences. When exclusively assessing cases with tophi, ultrasound and DECT demonstrated similar consistency in the medial, lateral and anterior views (κ = 0.633, 0.712, and 0.400, respectively), with statistically significant differences. In the intra-articular region, the consistency was reduced (κ = 0.237), and the differences were statistically significant. CONCLUSION: Ultrasound and DECT are effective methods to detect MSU deposition in gout of the knee. However, the consistency between the two techniques varies in different anatomical locations. Clinical assessment should be tailored based on the specific anatomical position. DECT is advantageous for the evaluation of intra-articular MSU deposits, while ultrasound is more sensitive for the early detection of scattered MSU deposits.

Overexpressing CsSABP2 enhances tolerance to Huanglongbing and citrus canker in C. sinensis.

Huanglongbing (HLB) and citrus canker, arising from Candidatus Liberibacter asiaticus (CaLas) and Xanthomonas citri pv. Citri (Xcc), respectively, have been imposing tremendous losses to the global citrus industry. Systemic acquired resistance (SAR) has been shown to be crucial for priming defense against pathogen in citrus. Salicylic acid (SA) binding protein 2 (SABP2), which is responsible for converting methyl salicylate (MeSA) to SA, is essential for full SAR establishment. Here, we characterized the functions of four citrus SABP2 genes (CsSABP2-1, CsSABP2-1V18A , CsSABP2-2 and CsSABP2-3) against HLB and citrus canker. In vitro enzymatic assay revealed that all four proteins had MeSA esterase activities, and CsSABP2-1 and CsSABP2-1V18A has the strongest activity. Their activities were inhibited by SA except for CsSABP2-1V18A. Four genes controlled by a strong promoter 35S were induced into Wanjincheng orange (Citrus sinensis Osbeck) to generate transgenic plants overexpressing CsSABP2. Overexpressing CsSABP2 increased SA and MeSA content and CsSABP2-1V18A had the strongest action on SA. Resistance evaluation demonstrated that only CsSABP2-1V18A had significantly enhanced tolerance to HLB, although all four CsSABP2s had increased tolerance to citrus canker. The data suggested the amino acid Val-18 in the active site of CsSABP2 plays a key role in protein function. Our study emphasized that balancing the levels of SA and MeSA is crucial for regulating SAR and conferring broad-spectrum resistance to HLB and citrus canker. This finding offers valuable insights for enhancing resistance through SAR engineering.

A Uniform Design Method Can Optimize the Combinatorial Parameters of Antimicrobial Photodynamic Therapy, Including the Concentrations of Methylene Blue and Potassium Iodide, Light Dose, and Methylene Blue's Incubation Time, to Improve Fungicidal Effects on Candida Species.

The optimal combinatorial parameters of antimicrobial photodynamic therapy (aPDT) mediated by methylene blue (MB) with the addition of potassium iodide (KI) against Candida species have never been defined. This study aimed to optimize the combinatorial parameters of aPDT, including the concentrations of MB (X1, 0.1-1.0 mM) and KI (X2, 100-400 mM), light dose (X3, 10-70 J/cm2), and MB's incubation time (X4, 5-35 min) for three Candida species. The best MB + KI-aPDT fungicidal effects (Y) against Candida albicans ATCC 90028 (YCa), Candida parapsilosis ATCC 22019 (YCp), and Candida glabrata ATCC 2950 (YCg) were investigated using a uniform design method. The regression models deduced using this method were YCa = 7.126 + 1.199X1X3 - 1.742X12 + 0.206X22 - 0.361X32; YCp = 10.724 - 0.867X1 - 1.497X2 + 0.560X3 + 1.298X22; and YCg = 0.892 - 0.956X1 + 2.296X3 + 1.299X42 - 3.316X3X4. The optimal combinatorial parameters inferred from the regression equations were MB 0.1 mM, KI 400 mM, a light dose of 20 J/cm2, and a 5-minute incubation time of MB for Candida albicans; MB 0.1 mM, KI 400 mM, a light dose of 70 J/cm2, and a 5-minute incubation time of MB for Candida parapsilosis; MB 0.1 mM, KI 100 mM, a light dose of 10 J/cm2, and a 35-minute incubation time of MB for Candida glabrata. The uniform design method can optimize the combinatorial parameters of aPDT mediated by MB plus KI to obtain the best aPDT fungicidal effects on Candida species, providing a new method to optimize the combinatorial parameters of aPDT for different pathogens in the future.

Function and molecular mechanism analysis of CaLasSDE460 effector involved in the pathogenesis of "Candidatus Liberibacter asiaticus" in citrus.

Citrus Huanglongbing (HLB), caused by Candidatus Liberibacter asiaticus (CaLas), is the most serious disease worldwide. CaLasSDE460 was previously characterized as a potential virulence factor of CaLas. However, the function and mechanism of CaLasSDE460 involved in CaLas against citrus is still elusive. Here, we showed that transgenic expression of CaLasSDE460 in Wanjincheng oranges (C. sinensis Osbeck) contributed to the early growth of CaLas and the development of symptoms. When the temperature increased from 25 °C to 32 °C, CaLas growth and symptom development in transgenic plants were slower than those in WT controls. RNA-seq analysis of transgenic plants showed that CaLasSDE460 affected multiple biological processes. At 25 °C, transcription activities of the "Protein processing in endoplasmic reticulum" and "Cyanoamino acid metabolism" pathways increased while transcription activities of many pathways decreased at 32 °C. 124 and 53 genes, separately annotated to plant-pathogen interaction and MAPK signaling pathways, showed decreased expression at 32 °C, compared with these (38 for plant-pathogen interaction and 17 for MAPK signaling) at 25 °C. Several important genes (MAPKKK14, HSP70b, NCED3 and WRKY33), remarkably affected by CaLasSDE460, were identified. Totally, our data suggested that CaLasSDE460 participated in the pathogenesis of CaLas through interfering transcription activities of citrus defense response and this interfering was temperature-dependent.

Phenylalanine-Arginine-ß-Naphthylamide Enhances the Photobactericidal Effect of Methylene Blue on Pseudomonas aeruginosa.

Objective: To investigate the effectiveness, dosing sequence, concentration, and mechanism of antimicrobial photodynamic inactivation (aPDI) using methylene blue (MB) plus phenylalanine-arginine-ß-naphthylamide (PAßN) against Pseudomonas aeruginosa. Methods: P. aeruginosa bacterial suspension was incubated with MB for different times (5-240 min), and then, 10 J/cm2 red light was irradiated. The efflux pump inhibitor (EPI) PAßN (10-100 µg/mL) was combined with MB (1-20 µM) in different sequences (PAßN-first, PAßN+MB, PAßN-after). Colony-forming units were then determined by serial dilution. Results: Using MB 10 µM plus 10 J/cm2, the killing effect of MB-aPDI on P. aeruginosa increased first and then decreased with longer incubation time. The killing effect of MB+PAßN-aPDI on P. aeruginosa was better than that of MB-aPDI (p < 0.05) by up to 2 logs. PAßN-first had the best killing effect, whereas PAßN-after had the worst killing effect. The killing effect increased with PAßN concentration and at 100 µg/mL reached 5.1 logs. Conclusions: The EPI PAßN enhanced the bactericidal effect of MB-aPDI on P. aeruginosa, especially when added before MB. It is proposed that MB is a substrate of the resistance-nodulation-division family efflux pump.

Exploration of influenza incidence prediction model based on meteorological factors in Lanzhou, China, 2014-2017.

Humans are susceptible to influenza. The influenza virus spreads quickly and behave seasonally. The seasonality and spread of influenza are often associated with meteorological factors and have spatio-temporal differences. Based on the influenza cases and daily average meteorological factors in Lanzhou from 2014 to 2017, this study firstly aimed to analyze the characteristics of influenza incidence in Lanzhou and the impact of meteorological factors on influenza activities. Then, SARIMA(X) models for the prediction were established. The influenza cases in Lanzhou from 2014 to 2017 was more male than female, and the younger the age, the higher the susceptibility; the epidemic characteristics showed that there is a peak in winter, a secondary peak in spring, and a trough in summer and autumn. The influenza cases in Lanzhou increased with increasing daily pressure, decreasing precipitation, average relative humidity, hours of sunshine, average daily temperature and average daily wind speed. Low temperature was a significant driving factor for the increase of transmission intensity of seasonal influenza. The SARIMAX (1,0,0)(1,0,1)[12] multivariable model with average temperature has better prediction performance than the university model. This model is helpful to establish an early warning system, and provide important evidence for the development of influenza control policies and public health interventions.

Transcription factor WRKY22 regulates canker susceptibility in sweet orange (Citrus sinensis Osbeck) by enhancing cell enlargement and CsLOB1 expression.

Pathological hypertrophy (cell enlargement) plays an important role in the development of citrus canker, but its regulators are largely unknown. Although WRKY22 is known to be involved in pathogen-triggered immunity and positively regulates resistance to bacterial pathogens in Arabidopsis, rice and pepper, the CRISPR/Cas9-mediated partial knockout of CsWRKY22 improves resistance to Xanthomonas citri subsp. citri (Xcc) in Wanjincheng orange (Citrus sinensis Osbeck). Here, we demonstrate that CsWRKY22 is a nucleus-localized transcriptional activator. CsWRKY22-overexpressing plants exhibited dwarf phenotypes that had wrinkled and thickened leaves and were more sensitive to Xcc, whereas CsWRKY22-silenced plants showed no visible phenotype changes and were more resistant to Xcc. Microscopic observations revealed that the overexpression of CsWRKY22 increased cell size in the spongy mesophyll. Transcriptome analysis showed that cell growth-related pathways, such as the auxin and brassinosteroid hormonal signaling and cell wall organization and biogenesis pathways, were significantly upregulated upon CsWRKY22 overexpression. Interestingly, CsWRKY22 activated the expression of CsLOB1, which is a key gene regulating susceptibility to citrus canker. We further confirmed that CsWRKY22 bound directly to the W-boxes just upstream of the transcription start site of CsLOB1 in vivo and in vitro. We conclude that CsWRKY22 enhances susceptibility to citrus canker by promoting host hypertrophy and CsLOB1 expression. Thus, our study provides new insights into the mechanism regulating pathological hypertrophy and the function of WRKY22 in citrus.

Overexpression of a "Candidatus Liberibacter Asiaticus" Effector Gene CaLasSDE115 Contributes to Early Colonization in Citrus sinensis.

Huanglongbing (HLB), caused by "Candidatus liberibacter asiaticus" (CaLas), is one of the most devastating diseases in citrus but its pathogenesis remains poorly understood. Here, we reported the role of the CaLasSDE115 (CLIBASIA_05115) effector, encoded by CaLas, during pathogen-host interactions. Bioinformatics analyses showed that CaLasSDE115 was 100% conserved in all reported CaLas strains but had sequence differences compared with orthologs from other "Candidatus liberibacter." Prediction of protein structures suggested that the crystal structure of CaLasSDE115 was very close to that of the invasion-related protein B (IalB), a virulence factor from Bartonella henselae. Alkaline phosphatase (PhoA) assay in E. coli further confirmed that CaLasSDE115 was a Sec-dependent secretory protein while subcellular localization analyses in tobacco showed that the mature protein of SDE115 (mSDE115), without its putative Sec-dependent signal peptide, was distributed in the cytoplasm and the nucleus. Expression levels of CaLasSDE115 in CaLas-infected Asian citrus psyllid (ACP) were much higher (∼45-fold) than those in CaLas-infected Wanjincheng oranges, with the expression in symptomatic leaves being significantly higher than that in asymptomatic ones. Additionally, the overexpression of mSDE115 favored CaLas proliferation during the early stages (2 months) of infection while promoting the development of symptoms. Hormone content and gene expression analysis of transgenic plants also suggested that overexpressing mSDE115 modulated the transcriptional regulation of genes involved in systemic acquired resistance (SAR) response. Overall, our data indicated that CaLasSDE115 effector contributed to the early colonization of citrus by the pathogen and worsened the occurrence of Huanglongbing symptoms, thereby providing a theoretical basis for further exploring the pathogenic mechanisms of Huanglongbing disease in citrus.

Antimicrobial photodynamic therapy for oral Candida infection in adult AIDS patients: A pilot clinical trial.

BACKGROUND: Antimicrobial photodynamic therapy (aPDT) using methylene blue (MB) plus potassium iodide (KI) has been shown to be effective in killing Candida albicans in many in vitro and in vivo studies, however, there are limited reports of clinical investigations. This study aimed to explore the clinical application of aPDT with MB plus KI for the treatment of oral infection caused by C. albicans in adult acquired immune deficiency syndrome (AIDS) patients. METHODS: A total of 21 adult AIDS patients with C. albicans oral candidiasis were divided into two groups according to MB concentration and received two consecutive aPDT treatments. Immediately before and after the aPDT treatments, C. albicans yeast isolates were recovered to measure the colony-forming units per mL (CFU/mL), biofilm formation, and to analyze the 25S rDNA genotype. Patients were assessed for the clinical recovery of oral lesions and improvement of symptoms. RESULTS: The Log10 CFU/mL of C. albicans decreased significantly after the second aPDT but not the first aPDT. There was no significant difference between the two MB concentrations. Both aPDT protocols decreased the oral lesions and clinical symptoms with no significant difference after 2-fraction aPDT. The biofilm formation of C. albicans isolates did not change before and after aPDT. The killing efficiency of 2-fraction-aPDT was not associated with either biofilm formation or 25S rDNA genotype. CONCLUSIONS: Two-fraction-aPDT with MB plus KI could reduce the number of viable C. albicans fungal cells and improve the clinical symptoms of oral candidiasis in adult AIDS patients, regardless of the biofilm formation or 25S rDNA genotype of infected C. albicans isolates.