Expanding the Structural Diversity of Tubulin-Targeting Agents: Development of Highly Potent Benzimidazoles for Treating Fungal Diseases.

Benzimidazoles, the representative pharmacophore of fungicides, have excellent antifungal potency, but their simple structure and single site of action have hindered their wider application in agriculture. In order to extend the structural diversity of tubulin-targeted benzimidazoles, novel benzimidazole derivatives were prepared by introducing the attractive pyrimidine pharmacophore. 2-((6-(4-(trifluoromethyl)phenoxy)pyrimidin-4-yl)thio)-1H-benzo[d]imidazole (A25) exhibited optimal antifungal activity against Sclerotinia sclerotiorum (S. s.), affording an excellent half-maximal effective concentration (EC50) of 0.158 µg/mL, which was higher than that of the reference agent carbendazim (EC50 = 0.594 µg/mL). Pot experiments revealed that compound A25 (200 µg/mL) had acceptable protective activity (84.7%) and curative activity (78.1%), which were comparable with that of carbendazim (protective activity: 90.8%; curative activity: 69.9%). Molecular docking displayed that multiple hydrogen bonds and π-π interactions could be formed between A25 and ß-tubulin, resulting in a stronger bonding effect than carbendazim. Fluorescence imaging revealed that the structure of intracellular microtubules can be changed significantly after A25 treatment. Overall, these remarkable antifungal profiles of constructed novel benzimidazole derivatives could facilitate the application of novel microtubule-targeting agents.

Dynamic of centromere associated RNAs and the centromere loading of DNA repair proteins in growing oocytes.

Mammalian centromeres are generally composed of dispersed repeats and the satellites such as α-satellites in human and major/minor satellites in mouse. Transcription of centromeres by RNA polymerase II is evolutionary conserved and critical for kinetochore assembly. In addition, it has been found that the transcribed satellite RNAs can bind DNA repair proteins such as MRE11 and PRKDC, and excessively expressed satellite RNAs could induce genome instability and facilitate tumorigenesis. During the maturation of female oocyte, centromeres are critical for accurate segregation of homologous chromosomes and sister chromatids. However, the dynamics of oocyte centromere transcription and whether it associated with DNA repair proteins are unknown. In this study, we found the transcription of centromeres is active in growing oocytes but it is silenced when oocytes are fully grown. DNA repair proteins like Mlh1, Mre11 and Prkdc are found associated with the minor satellites and this association can be interfered by RNA polymerase II inhibitor α-amanitin. When the growing oocyte is in vitro matured, Mlh1/Mre11/Prkdc foci would release from centromeres to the ooplasm. If the oocytes are treated with Mre11 inhibitor Mirin, the meiosis resumption of growing oocytes with Mre11 foci can be suppressed. These data revealed the dynamic of centromeric transcription in oocytes and its potential association with DNA repair proteins, which provide clues about how oocytes maintain centromere stability and assemble kinetochores.

Exploiting Natural Maltol for Synthesis of Novel Hydroxypyridone Derivatives as Promising Anti-Virulence Agents in Bactericides Discovery.

Anti-infection strategies based on suppression of bacterial virulence factors represent a crucial direction for the development of new antibacterial agents to address the resistance triggered by traditional drugs'/pesticides' bactericidal activity. To identify and obtain more effective and diverse molecules targeting virulence, we prepared a series of 3-hydroxy-2-methyl-1-pyridin-4-(1H)-one derivatives and evaluated their antibacterial behaviors. Compound B6 exhibited the highest bioactivity, with half-maximal effective concentration (EC50) values ranging fro9m 10.03 to 30.16 µg mL-1 against three plant pathogenic bacteria. The antibacterial mechanism showed that it could considerably reduce various virulence factors (such as extracellular enzymes, biofilm, and T3SS effectors) and inhibit the expression of virulence factor-related genes. In addition, the control efficiency of compound B6 against rice bacterial leaf blight at 200 µg mL-1 was 46.15-49.15%, and their control efficiency was improved by approximately 12% after the addition of pesticide additives. Thus, a new class of bactericidal candidates targeting bacterial virulence factors was developed for controlling plant bacterial diseases.

Effects of different viscoelastic agents on corneal endothelium of patients after phacoemulsification and foldable IOL implantation / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To analyze the effects of dual viscoelastic agents DisCoVisc and sodium hyaluronate on corneal endothelium of patients after phacoemulsification and foldable intraocular lens(IOL)implantation.METHODS: A total of 247 patients(285 eyes)with cataract treated in Jingmen Aier Eye Hospital between June 2017 and December 2019 were selected, and they were divided into DisCoVisc group(123 cases, 141 eyes)and sodium hyaluronate group(124 cases, 144 eyes)by random number table method. Both groups were treated with phacoemulsification and foldable IOL implantation. DisCoVisc and 1.7% sodium hyaluronate were used as viscoelastic agents in DisCoVisc group and sodium hyaluronate group, respectively. The two groups were compared in terms of intraoperative ultrasound time(UST), cumulative dissipated energy(CDE), time for aspiration of viscoelastic agents after IOL implantation, corneal edema at 1d, 1wk, 1 and 3mo after operation, corneal endothelial cell density(ECD)and ECD loss rates before operation and at 3mo after operation, coefficient variation of corneal endothelial cell size(CV), percentage of corneal hexagonal endothelial cells(6A), intraocular pressure, the proportions of patients with uncorrected visual acuity ≥0.5 and central corneal thickness(CCT)values before and after operation at 1d, 1wk and 1mo.RESULTS:There was no statistically significant difference between the two groups in UST, CDE, aspiration time of viscoelastic agents(P >0.05)or corneal edema both rate on day 1 after operation(P>0.05). Corneal edema disappeared at 1 wk after operation. The ECD loss rate in DisCoVisc group was significantly lower than that in sodium hyaluronate group at 3mo after operation(P<0.05). Intraocular pressure, the proportion of patients with uncorrected visual acuity ≥0.5 and CCT values showed no statistically significant differences between the two groups before and after operation at 1d, 1wk and 1mo(P >0.05).CONCLUSION: DisCoVisc, as the viscoelastic agent in phacoemulsification and foldable IOL implantation for patients with Emery-Little grade Ⅱ-Ⅲ lens nucleus hardness, can better protect the patients' corneal endothelium.

Fabrication of Versatile Pyrazole Hydrazide Derivatives Bearing a 1,3,4-Oxadiazole Core as Multipurpose Agricultural Chemicals against Plant Fungal, Oomycete, and Bacterial Diseases.

Developing multipurpose agricultural chemicals is appealing in crop protection, thus eventually realizing the reduction and efficient usage of pesticides. Herein, an array of versatile pyrazole hydrazide derivatives bearing a 1,3,4-oxadiazole core were initially synthesized and biologically evaluated the antifungal, antioomycetes, and antibacterial activities. In addition, the pyrazole ring was replaced by the correlative pyrrole, thiazole, and indole scaffolds to extend the molecular diversity. The results showed that most of these hybrid compounds were empowered with multifunctional bioactivities, which are exemplified by compounds a1-a6, b1-b3, b7, b10, b13, and b18. For the antifungal activity, the minimal EC50 values could afford 0.47 (a2), 1.05 (a2), 0.65 (a1), and 1.32 µg/mL (b3) against the corresponding fungi Gibberella zeae (G. z.), Fusarium oxysporum, Botryosphaeria dothidea, and Rhizoctonia solani. In vivo pot experiments against corn scab (caused by G. z.) revealed that the compound a2 was effective with protective and curative activities of 90.2 and 86.3% at 200 µg/mL, which was comparable to those of fungicides boscalid and fluopyram. Further molecular docking study and enzymatic activity analysis (IC50 = 3.21 µM, a2) indicated that target compounds were promising succinate dehydrogenase inhibitors. Additionally, compounds b2 and a4 yielded superior anti-oomycete and antibacterial activities toward Phytophora infestins and Xanthomonas oryzae pv. oryzae with EC50 values of 2.92 and 8.43 µg/mL, respectively. In vivo trials against rice bacterial blight provided the control efficiency within 51.2-55.3% (a4) at 200 µg/mL, which were better than that of bismerthiazol. Given their multipurpose characteristics, these structures should be positively explored as agricultural chemicals.

Zinc Absorption from Representative Diet in a Chinese Elderly Population Using Stable Isotope Technique.

OBJECTIVE: To determine the dietary zinc absorption in a Chinese elderly population and provide the basic data for the setting of zinc (Zn) recommended nutrient intakes (RNI) for Chinese elderly people. METHODS: A total of 24 elderly people were recruited for this study and were administered oral doses of 3 mg 67Zn and 1.2 mg dysprosium on the fourth day. The primary macronutrients, energy, and phytic acid in the representative diet were examined based on the Chinese National Standard Methods. Fecal samples were collected during the experimental period and analyzed for zinc content, 67Zn isotope ratio, and dysprosium content. RESULTS: The mean (± SD) zinc intake from the representative Chinese diet was 10.6 ± 1.5 mg/d. The phytic acid-to-zinc molar ratio in the diet was 6.4. The absorption rate of 67Zn was 27.9% ± 9.2%. The RNI of zinc, which were calculated by the absorption rate in elderly men and women, were 10.4 and 9.2 mg/d, respectively. CONCLUSION: This study got the dietary Zn absorption in a Chinese elderly population. We found that Zn absorption was higher in elderly men than in elderly women. The current RNI in elderly female is lower than our finding, which indicates that more attention is needed regarding elderly females' zinc status and health.

[Construction of P2Y6 constitutive knock down breast cancer cell line and evaluation of its proliferation].

AIM: To construct the constitutive P2Y6 knock down breast cancer cell line with shRNA technology and provide a basis for discovering how P2Y6 regulates tumorigenesis and progression in breast cancer. METHODS: The paired oligo nucleotides targeting human P2Y6 gene were synthesized and annealed into linear PLKO lentiviral vector digested by EcoRI and AgeI. The recombined vector which was identified by double digest with EcoRI and NdeI and DNA sequencing was packaged in 293T cells together with psPAX2 and pMD2.G. Virus in culture supernatant was concentrated, three recombinant vectors were transfected into BT549 cells, and the constitutive P2Y6 knock down cells were selected by puromycin. The efficiency of RNA interference was detected by RT-PCR and Western blotting. MTS assay was used to detect the influence of P2Y6 on breast cancer cells'proliferation. RESULTS: The inserted sequence was proven to be correct by DNA sequencing. After stable transfection of P2Y6 shRNA, the expression of P2Y6 in BT549 cells was decreased obviously in both protein and mRNA level. But no obvious influence on proliferation was found in P2Y6 gene knock down cells. CONCLUSION: The constitutive P2Y6 knock down cell line was successfully constructed in BT549 cells. Whereas, no obvious correlation was found between the expression of P2Y6 and breast cancer cell proliferation.

Evaluation of the expert patient program in a Chinese population with permanent colostomy.

BACKGROUND: A colostomy can negatively impact patients' quality of life. Recent research findings suggest that self-efficacy and psychosocial adjustment are positively associated with quality of life, but there are few research reports about the self-efficacy and psychosocial adjustment of patients with a permanent colostomy. OBJECTIVES: The objective of the study was to evaluate the effect of a 3-week Expert Patient Program (EPP) on colostomy knowledge, stoma care self-efficacy, self-management, and psychosocial adjustment in Chinese patients who had a permanent colostomy. METHODS: Eleven patients with permanent colostomy volunteered to be trained to be expert patients. Eighty-one patients with permanent colostomy were recruited and participated in the 3-week EPP courses. Questionnaires with items about knowledge on colostomy, stoma care self efficacy, self-management, and psychosocial adjustment were administered before and at 4 weeks following the EPP courses. Each EPP participant also completed a verbal interview at the end of the program. RESULTS: The EPP participants demonstrated statistically significant improvement (P < .01) in knowledge, self-efficacy, self-management, and psychosocial adjustment to colostomy (mean [SD]: 51.89 [11.45] vs 89.22 [19.90], 71.56 [5.93] vs 85.61 [14.32], 125.44 [19.27] vs 140.78 [15.34], 47.59 [9.64] vs 53.37 [10.68], respectively). Most participants gave positive ratings to the EPP. CONCLUSIONS: This study showed that the EPP was applied effectively in patients with permanent colostomy. IMPLICATIONS FOR PRACTICE: This study highlights the importance of psychosocial support for patients with permanent colostomy. It offers a model of self-help practice in colostomy patients, supported by community that can benefit more patients with permanent colostomy.