First Report of Neofusicoccum parvum Causing Seedpod Blight on Lotus (Nelumbo nucifera) in China.

Lotus (Nelumbo nucifera Gaertn.) is a widely cultivated plant in China, and the fruit lotus variety has a high economic value attributed to the exquisite flavor of its fresh seeds. During the summer of 2023, an unidentified blight was observed affecting lotus seedpods in Jiande City, Zhejiang province, with approximately 65% of seedpods impacted in a 130-hectare area. The initial symptoms included dark purple spots on the lotus seedpod surface, which gradually expanded over time. After 5 to 7 days, the entire seedpod turned black, withering, and rendering the lotus seeds inedible. To identify the causal agent, tissues from symptomatic seedpods were excised and disinfected in 75% ethanol for 60 s, and washed twice in sterile distilled water. The disinfected symptomatic tissues (5 × 5 mm) were plated on potato dextrose agar (PDA), incubated at 25 ℃, transferred hyphal tips to obtain pure isolates after 3 days. Fungal colonies exhibiting Botryosphaeriaceae morphology were isolated from 33% of the samples (n = 15). Pure cultures were grown on PDA for both morphological and molecular identification. The colonies displayed a white aerial mycelium, turning olivaceous grey after 7 days. Pycnidia were produced within 3 weeks on PDA with added sterilized healthy lotus seedpod pieces on the surface. Conidia were hyaline, unicellular, ellipsoidal, 12.65 to 20.72 × 3.92 to 9.38 µm in size (mean 16.67 × 6.24 µm, n = 100). To determine the fungal species, genomic DNA was extracted from one representative isolate (ZJUP1112-1), to amplify four gene loci through polymerase chain reactions (PCR): rDNA internal transcribed spacer (ITS) with primers ITS1/ITS4, rDNA large subunit (LSU) with LR0R/LR5, the translation elongation factor 1-alpha gene (tef1) with EF1-728F/EF1-986R, and ß-tubulin gene (tub2) with Bt2a/Bt2b. The PCR products were Sanger sequenced in Zhejiang Shangya biotechnology co., LTD, and the resulting sequences were assembled and deposited in GenBank (ITS: OR740546; LSU: OR740547; tef1: OR776996; tub2: OR776997). BLAST searches indicated the highest nucleotide sequence identity with the reference strains of Neofusicoccum parvum CMW 9081 (ITS: 98.8%, AY236943; LSU: 100%, AY928045; tef1: 99.6%, AY236888; tub2: 99.3%, AY236917). Multi-locus phylogenetic analyses revealed that isolate ZJUP1112-1 formed a highly supported clade with N. parvum. Pathogenicity tests were performed on healthy lotus seedpods using mycelial plugs (5 mm diameter) from actively growing colonies of ZJUP1112-1 that were placed onto the front and side of the seedpods (6 each). Controls received PDA plugs. Treated seedpods were wrapped with parafilm and incubated at 25 ℃ and the experiment was repeated three times. After 5 days, dark purple lesions were observed on the inoculated seedpods, whereas controls remained symptomless. The same isolate was recovered from the margin of resulting lesions and confirmed by morphology, thus fulfilling Koch's postulates. N. parvum is a polyphagous pathogen causing blights and fruit rot on multiple economically important fruit crops, such as cacao (Puig et al. 2019), walnut (Chen et al. 2019), pistachio (Lopez-Moral et al. 2020), chestnut (Seddaiu et al. 2021), blueberry (Spetik et al. 2023) and mango (Polizzi et al. 2022), among others. To the best of our knowledge, this is the first report of N. parvum causing seedpod blight on lotus seedpods in China, which contributes to a better understanding of the pathogens affecting this plant species in China.

[Source Analysis and Risk Assessment of Heavy Metals in Soil of County Scale Based on PMF Model].

This study explores the pollution characteristics, risks, and sources of heavy metals in small-scale areas. Rongcheng District, Jieyang City, Guangdong Province was considered as the study area and enrichment factor (EF), pollution load index (PLI), potential ecological risk index (RI), and US EPA health risk assessment model were used to evaluate its environmental risk. Moreover, the source apportionment of heavy metals was analyzed through correlation analysis, the characteristic of spatial distribution, and a PMF model. The results showed that the mean concentrations of ω(Cr), ω(Hg), ω(As), ω(Pb), ω(Ni), ω(Cd), ω(Cu), and ω(Zn) were 54.87, 0.25, 8.35, 56.00, 15.38, 0.35, 30.56, and 124.23 mg·kg-1, respectively. The mean concentrations of all elements exceeded the local soil background value. In terms of EF level, Cr, As, Pb, and Ni showed negligible accumulation; Zn and Cu showed minor accumulation; and Hg and Cd showed moderate accumulation. The mean value of the pollution load index was 2.37, with a severe pollution level, and the eight elements were in different pollution levels. In total, the study region suffered severe ecological risk, Hg and Cd presented strong ecological risk, and other elements presented slight ecological risk. The non-carcinogenic risks under the three exposure paths were within the acceptable level. The carcinogenic risks (CR) of adults and children were 9.81E-05 and 5.59E-04, respectively, and Cr and As were the main contributors of CR. The results showed that the four sources of heavy metals were transportation sources (37.02%), parent material sources (18.53%), atmospheric deposition sources (26.49%), and industrial sources (17.96%).

Performance of Creatinine- and Cystatin C-Based Equations for Glomerular Filtration Rate Estimation in HIV-1-Infected Individuals Receiving Dolutegravir + Tenofovir Disoproxil Fumarate + Lamivudine as Initial Antiretroviral Therapy: A Retrospective Observational Study.

BACKGROUND: Dolutegravir (DTG) may inhibit organic cation transporter-2 in renal tubules and elevate serum creatinine levels without true renal function deterioration. There are scarce data on the glomerular filtration rate (GFR) equations in Chinese HIV patients with DTG/TDF/3 TC. This retrospective study was to evaluate the changes in creatinine- and cystatin C-based estimated GFR values among Chinese adult HIV patients with initial highly active antiretroviral therapy (HAART) with DTG/TDF/3 TC for 48 weeks. SETTINGS: Retrospective analysis of Chinese adult HIV patients who received initial HAART with DTG/TDF/3 TC between January 2016 and May 2021. METHODS: We compared the changes in creatinine-based and cystatin C-based estimated GFR before and 48 weeks after HAART to determine the optimal tool for renal function estimation. RESULTS: Ninety-five patients [91.58% men; median age, 34 years (interquartile range, 29-44 years)] were enrolled. The median baseline CD4+ cell count was 185 cells/µL [interquartile range, 53-303 cells/µL] and the median baseline viral load was 148,000 copies/mL [interquartile range, 26,800-596,000 copies/mL]. The estimated GFR at 4, 12, and 24 weeks was significantly different than at baseline (P < 0.05) using all 4 equations. Only the GFR estimated using the CKD-EPIcreat and CKD-EPIcys equations was significantly different at 48 weeks of treatment (P < 0.05). The modification of diet in renal disease-based decline of GFR incidence at 4, 12, 24, and 48 weeks of treatment (21.5%, 24.21%, 33.68% and 38.95%, respectively) was higher than that calculated using the other 3 GFR equations. CONCLUSIONS: The modification of diet in renal disease equation may not be optimal for Chinese HIV-1-infected adults receiving DTG + TDF + 3 TC as the initial antiviral therapy. Clinicians must carefully choose the GFR equation for patients with HIV/hepatitis B virus coinfection.

Erythrocyte Membrane Cloaked Curcumin-Loaded Nanoparticles for Enhanced Chemotherapy.

In this study, curcumin-loaded porous poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) were prepared and surface modified with red blood cell membranes (RBCM) to yield biomimetic RBCM-p-PLGA@Cur NPs. The NPs displayed a visible cell-membrane structure at their exterior and had a uniform size of 162 ± 3 nm. In vitro studies showed that drug release from non-porous PLGA NPs was slow and that much of the drug remained trapped in the NPs. In contrast, release was accelerated from the porous PLGA NPs, and after the RBCM coating, a sustained release over 48 h was obtained. Confocal microscopy and flow cytometry results revealed that the RBCM-p-PLGA NPs led to a greater cellular uptake by H22 hepatocarcinoma cells than the uncoated analogue NPs, but could avoid phagocytosis by macrophages. The drug-free formulations were highly biocompatible, while the drug-loaded systems were effective in killing cancer cells. RBCM-p-PLGA@Cur NPs possess potent anti-tumor activity in a murine H22 xenograft cancer model (in terms of reduced tumor volume and mass, as well as inducing apoptosis of tumor cells), and have no observable systemic toxicity. Overall, our study demonstrates that the use of the RBCM to cloak nanoscale drug delivery systems holds great promise for targeted cancer treatment, and can ameliorate the severe side effects currently associated with chemotherapy.

One-pot synthesis of thermosensitive glycopolymers grafted gold nanoparticles and their lectin recognition.

Thermosensitive glucose-functionalized glycopolymers grafted gold nanoparticles (Glyco@GNPs) with good colloidal stability and thermosensitive in aqueous solution were fabricated by reversible addition-fragmentation chain transfer (RAFT) mediated one-pot synthesis. The formation of core-shell morphology with about a 60 nm gold core in diameter and a glycopolymer shell of about 80 nm in thickness was indicated by transmission electron microscopy (TEM). The recognition ability of the Glyco@GNPs toward lectin concannavalin A (Con A) was verified by ultraviolet-visible spectroscopy and dynamic light scattering (DLS). The good cytocompatibility of the glycopolymers and Glyco@GNPs was proven by MTT assay on L-929 cells. Glyco@GNPs could effectively inhibit hepatoma cells SMMC-7721 growth after recognizing Con A was also proved by MTT assay and flow cytometry assay.

Prediction value of unmeasurable MR enhancement at early stage after gross-total resection on the survival state of patients with high-grade glioma.

PROPOSE: To explore the value of unmeasurable enhancement pattern of residual cavity in predicting survival at early stage after gross-total resection in high-grade glioma (HGG) patients. METHODS: This retrospective study enrolled consecutive 51 HGG patients with unmeasurable enhancement who underwent gross-total resection followed by concurrent chemoradiotherapy and adjuvant chemotherapy. We evaluated the enhancement patterns of residual cavity on contrast-T1WI made within 1 month after tumor resection (20 ± 3 days). The survival state of different enhancement was compared. RESULTS: Thin-linear, thick-linear and nodular enhancement were observed in 22 patients (43%), 10 patients (20%), and 19 patients (37%), respectively. The progression-free survival of patients with thin-linear (487, 151-887 days) was longer than those patients with thick-linear (277, 133-573 days), and nodular enhancement (210, 120-765 days) (P = 0.002). The overall survival of patients with thin-linear (774, 457-1343 days) was longer than those with thick-linear (462, 320-678 days), and nodular enhancement (326, 234-1393 days) (P = 0.002). There was no significant difference of orthogonal value between thick-linear and nodular enhancement (0.854), neither between grade III and IV with same enhancement patterns (P = 0.540, P = 0.720). CONCLUSIONS: The unmeasurable enhancement patterns in HGG patients within 1 month after gross-total resection, which might be better than the grade of tumor, holds a potential marker in survival state.

Increasing FLAIR signal intensity in the postoperative cavity predicts progression in gross-total resected high-grade gliomas.

To evaluate the prognostic value of fluid-attenuated inversion recovery (FLAIR) signal intensity of postoperative cavity on progression free survival (PFS) and overall survival (OS) in patients with high-grade gliomas (HGG). This study retrospectively enrolled 45 consecutive HGG patients. These patients had chemoradiotherapy after gross-total resection of tumors. Quantitative analysis of the FLAIR signal intensity in postoperative cavity and background was made. We evaluated the threshold value, accuracy, sensitivity, specificity, and survival state with this technique. The patients who progressed and patients who did not progress were 33 and 12 cases separately. The ratio of postoperative cavity and background (C-B) on FLAIR sequence in patients who progressed was higher than that of patients who did not progress (P = 0.014). The PFS of the patients who progressed was shorter than that of patients who did not progress (P = 0.008). The area under ROC curve, threshold, sensitivity, specificity of C-B ratio for predicting tumor progression were 0.875, 62.3, 69.7, 0.84, and 0.50% respectively. The PFS of lower signal group was much longer than that of higher signal group (P = 0.004). The OS of the patients with higher signal was shorter than that of patients with lower signal (P = 0.034). The increase of gray value of FLAIR in postoperative cavity may be used as an imaging marker for predicting tumor progression.

A three-channel fluorescent probe that distinguishes peroxynitrite from hypochlorite.

A novel fluorescent probe for peroxynitrite, PN(600), was rationally designed on the basis of a unique fluorophore assembly approach. PN(600) is a green-emitting coumarin derivative. Upon oxidation by peroxynitrite, PN(600) is transformed into a highly fluorescent red-emitting resorufin derivative via an orange-emitting intermediate. This three-channel signaling capability enables PN(600) to differentiate peroxynitrite from other reactive oxygen and nitrogen species, including hypochlorite and hydroxyl radical. Moreover, PN(600) is membrane-permeable and compatible with common TRITC filter sets and displays low cytotoxicity. Therefore, PN(600) is a promising candidate for in vitro peroxynitrite imaging.

Preparation of chitosan-polyaspartic acid-5-fluorouracil nanoparticles and its anti-carcinoma effect on tumor growth in nude mice.

AIM: To prepare chitosan-polyaspartic acid-5-fluorouracil (CTS-Pasp-5Fu) nanoparticles and investigate its anti-carcinoma effect and toxicity. METHODS: CTS-Pasp-5Fu nanoparticles were synthesized by ionic gelatification. Male BABL/c nude mice were injected with SGC-7901 gastric carcinoma cell line mass to establish a human gastric carcinoma model. They were randomly allocated into 4 groups: CTS-Pasp-5Fu (containing 5-Fu 1.25 mg/kg), 5-Fu (1.25 mg/kg), CTS-Pasp and normal saline groups. Tumor weight was measured and assay of colony forming unit-granulocyte and macrophage (CFU-GM) was performed. The structural change of cells and tissues was observed and the Bax and Bcl-2 genes were detected. RESULTS: Compared with normal saline, the inhibition rates of tumor growth for the CTS-Pasp, 5-Fu and CTS-Pasp-5Fu groups were 5.58%, 58.69% and 70.82%, respectively. The tumor inhibition rates for the CTS-Pasp, 5-Fu and CTS-Pasp-5Fu groups were 5.09%, 65.3% and 72.79%, respectively. There was a significant decrease in the number of CFU-GM formation and increase of total bilirubin, and alanine aminotransferase in the 5-Fu group, but no change in those of the other three groups. There was no change in white blood cell count and creatinine among the four groups. Pathological section of liver and nephridial tissues showed that the damage in the 5-Fu group was more severe than that in the CTS-Pasp-5Fu group. 5-Fu and CTS-Pasp-5Fu groups could both down-regulate the Bcl-2 expression and up-regulate the Bax expression to different extent, and the accommodate effect of CTS-Pasp-5Fu was more obvious than 5-Fu. CONCLUSION: The tumor inhibition rate of CTS-Pasp-5Fu nanoparticles is much higher than that of 5-Fu alone.

Nanoparticles based on the complex of chitosan and polyaspartic acid sodium salt: preparation, characterization and the use for 5-fluorouracil delivery.

New nonstoichiometric polyelectrolyte complex nanoparticles were prepared based on chitosan (CS) and polyaspartic acid sodium salt (PAsp). The physicochemical properties of the complexes were investigated by means of turbidity, dynamic light scattering, transmission electron microscopy and zeta potential. The results indicated that the slow dropwise addition of chitosan into PAsp allowed to elaborate either anionic or cationic particles in the size range of 85-300 nm with proper CS and PAsp unit molar ratios. Investigation of structural changes during the addition of CS revealed that the microstructure of the nanoparticles depended strongly on the unit molar ratio of CS to PAsp. Nanoparticles containing a hydrophilic drug, 5-fluorouracil (5FU), were prepared by mixing and absorption method. In vitro and in vivo experiment indicated that the drug-loaded CS-PAsp nanoparticles presented a sustained release of 5FU compared to the 5FU solution and the areas under curve (AUC) were increased by about four times.