First Report of the Root-Knot Nematode Meloidogyne incognita on Bletilla striata Rchb.f. (Orchidaceae) in China.

Bletilla striata Rchb.f., is a perennial herbaceous bulbous plant known as the Chinese ground or hyacinth orchid classified in the Orchidaceae. It is native to southeast Asia and mainly distributed in China, Japan and northern Myanmar (He et al. 2017). It has the functions of astringent hemostasis and analgesia, and can also be used to treat traumatic bleeding, ulcers, swelling and chapped skin. Therefore, it occupies an important position in traditional Chinese medicine (Xu et al. 2019). In June 2023, three farmers in Mengzi (103.39°N, 23.21°E), Yunnan Province, China, observed that some Bletilla striata Rchb.f. plants grew poorly with small and chlorotic leaves (Figure 1 A). We suspected that these symptoms were caused by root-knot nematode infection, but the galls on the roots were small and inconspicuous (Figure 1 A). The presence of nematode females in both the galled regions and the normal roots (Figure 1 B), revealed by fuchsin staining (Byrd et al. 1983), indicated that the symptoms were probably caused by root-knot nematode infection. To estimate the incidence rates, we randomly selected 100 B. striata Rchb.f. plants from each of five fields representing a total area of 3000 m2. In these fields, the occurrence of stained root-knot nematodes were 19.3%, 17%, 18.3%, 15%, and 13%, respectively. The gall rating of the infected plants in the B striata Rchb.f. samples collected from the five fields was 2 (rating scale of 0 to 5). Females (n=20), second-stage juveniles (J2s, n=20) and egg masses (n=20) were extracted and collected from roots and soil for morphological and molecular identification. The females had a white, pyriform body and their perineal patterns exhibited a high and square dorsal arch, lacking distinct lateral line (Figure 1. C & D). Measurements of females (n = 20) were: body length (BL) = 708.64±89.6 µm (554.36 to 844.51 µm); maximum body width (BW) = 461.73±47.44 µm (365.25 to 561.49 µm); stylet length (ST) = 15.49±3.15 µm (10.55 to 19.78 µm); and distance from dorsal esophageal gland opening to the stylet knobs (DGO) = 3.33±0.27 µm (2.77 to 3.93 µm). Measurements of J2s (n=20) were BL = 417.7±47.67 µm (342.16 to 499.68 µm); BW = 15.74±2.66 µm (11.05 to 25.63 µm); ST = 12.49±1.12 µm (10.19 to 15.02 µm); DGO = 2.64±0.59 µm (40.17 to 68.74 µm); tail length = 51.93±8.55 µm (10.43 to 27.22 µm); hyaline tail terminus = 18.23±3.99 µm (1.48 to 3.98 µm). These morphological features match the description of Meloidogyne incognita (Eisenback et al. 1981). To further confirm the species, we selected three infected plants from each field for molecular identification, the ITS region amplified using the primers 18S/26S (5'-TTGATTACGTCCCTGCCCTTT-3',5'-TTTCACTCGCCGTTACTAAGG-3') (Vrain et al. 1992) . A 729 bp PCR product of ITS region (accession nos. OR463907) was obtained from all infected plants. The amplicons from 18S/26S primer pair were sequenced and the sequences showed 95.29% homology with sequences of M. incognita (accession nos. MT209948.1). Moreover, a 835 bp DNA fragment (accession nos. OR469000) was obtained using the specific primers Mi-F/Mi-R (5'-GTGAGGATTCAGCTCCCCAG-3',5'-ACGAGGAACATACTTCTCCGTCC-3') for M. incognita (Meng et al. 2004), the sequence showed 99.28% homology with sequences of M. incognita (accession nos. ON416569). The morphological features and molecular data confirmed the identification of the root-knot nematode on B. striata Rchb.f. as M. incognita. To confirm the pathogenicity, ten healthy B. striata Rchb.f. seedlings were each inoculated with 500 freshly hatched J2s isolated from field Bletilla striata Rchb.f.. Five healthy seedlings without J2 inoculation were used as controls. At 60 days after inoculation, most of the inoculated plants exhibited similar symptoms to those initially observed by farmers in the field. On average, 1532 J2s were recovered from each inoculated plant, yielding a reproductive factor of 2.1. The gall rating for these inoculated plants was 2. Fuchsin staining revealed the presence of root-knot nematode females within the roots, with an average of 17 females detected per inoculated plant. No symptoms were observed in the control plants. This is the first report of M. incognita infecting B. striata Rchb.f. in China. M. incognita can cause severe infection and damage to some crops, resulting in serious economic losses (Eisenback, 2022). The growers need to take measures to prevent the spread of this nematode.

Small molecules targeting molecular chaperones for tau regulation: Achievements and challenges.

Abnormal post-translational modification of microtubule-associated protein Tau (MAPT) is a prominent pathological feature in Alzheimer's disease (AD). Previous research has focused on designing small molecules to target Tau modification, aiming to restore microtubule stability and regulate Tau levels in vivo. However, progress has been hindered, and no effective Tau-targeted drugs have been successfully marketed, which urgently requires more strategies. Heat shock proteins (HSPs), especially Hsp90 and Hsp70, have been found to play a crucial role in Tau maturation and degradation. This review explores innovative approaches using small molecules that interact with the chaperone system to regulate Tau levels. We provide a comprehensive overview of the mechanisms involving HSPs and their co-chaperones in the Tau regulation cycle. Additionally, we analyze small molecules targeting these chaperone systems to modulate Tau function. By understanding the characteristics of the molecular chaperone system and its specific impact on Tau, we aim to provide a perspective that seeks to regulate Tau levels through the manipulation of the molecular chaperone system and ultimately develop effective treatments for AD.

Design of Tracers in Fluorescence Polarization Assay for Extensive Application in Small Molecule Drug Discovery.

Development of fluorescence polarization (FP) assays, especially in a competitive manner, is a potent and mature tool for measuring the binding affinities of small molecules. This approach is suitable for high-throughput screening (HTS) for initial ligands and is also applicable for further study of the structure-activity relationships (SARs) of candidate compounds for drug discovery. Buffer and tracer, especially rational design of the tracer, play a vital role in an FP assay system. In this perspective, we provided different kinds of approaches for tracer design based on successful cases in recent years. We classified these tracers by different types of ligands in tracers, including peptide, nucleic acid, natural product, and small molecule. To make this technology accessible for more targets, we briefly described the basic theory and workflow, followed by highlighting the design and application of typical FP tracers from a perspective of medicinal chemistry.

Baricitinib Attenuates Bleomycin-Induced Pulmonary Fibrosis in Mice by Inhibiting TGF-ß1 Signaling Pathway.

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive interstitial lung disease with unknown etiology, high mortality and limited treatment options. It is characterized by myofibroblast proliferation and extensive deposition of extracellular matrix (ECM), which will lead to fibrous proliferation and the destruction of lung structure. Transforming growth factor-ß1 (TGF-ß1) is widely recognized as a central pathway of pulmonary fibrosis, and the suppression of TGF-ß1 or the TGF-ß1-regulated signaling pathway may thus offer potential antifibrotic therapies. JAK-STAT is a downstream signaling pathway regulated by TGF-ß1. JAK1/2 inhibitor baricitinib is a marketed drug for the treatment of rheumatoid arthritis, but its role in pulmonary fibrosis has not been reported. This study explored the potential effect and mechanism of baricitinib on pulmonary fibrosis in vivo and in vitro. The in vivo studies have shown that baricitinib can effectively attenuate bleomycin (BLM)-induced pulmonary fibrosis, and in vitro studies showed that baricitinib attenuates TGF-ß1-induced fibroblast activation and epithelial cell injury by inhibiting TGF-ß1/non-Smad and TGF-ß1/JAK/STAT signaling pathways, respectively. In conclusion, baricitinib, a JAK1/2 inhibitor, impedes myofibroblast activation and epithelial injury via targeting the TGF-ß1 signaling pathway and reduces BLM-induced pulmonary fibrosis in mice.

Application of improved high-risk sign board for older people inpatients: a retrospective study.

AIM: To observe the effect of the implementation of improved high-risk sign boards for older people inpatients. METHOD: The older people inpatients of the Department of Geriatrics at our hospital were selected as the research subjects and divided into two groups. The control group used the single-strip high-risk sign, and the intervention group used the improved double-layer card slot, push-pull integrated high-risk sign board (national patent). The sign-related nurse operation time, patient/attendant satisfaction, and high-risk-related adverse events were observed and compared between the two groups. RESULTS: After the adoption of the improved high-risk sign board, the nurse operation time was reduced from 94.3 ± 16.2 s to 53.9 ± 12.5 s, and patient/attendant satisfaction increased from 6.65 ± 0.38 points to 9.30 ± 0.52 points (P < 0.001). The incidence of high-risk-related adverse events decreased from 6.08 to 1.86%, but the difference was not statistically significant (χ2 = 3.675, P = 0.055). The implementation of the improved high-risk sign board can increase nursing efficiency and enhance the awareness of risk prevention in high-risk patients among nurses, older people inpatients, and attendants. CONCLUSION: The application of double-layer card slot and push-pull comprehensive high-risk identification card to older people inpatients can alert nurses, patients, and nursing staff more prominently, which can improve patient satisfaction, reduce installation time and reduce the incidence of adverse events to a certain extent.

Zanubrutinib attenuates bleomycin-induced pulmonary fibrosis by inhibiting the TGF-ß1 signaling pathway.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive and fatal interstitial lung disease with high mortality and limited treatment. Only two drugs are currently approved for the treatment of IPF, but both have limitations and neither drug could prolong survival time of patients. The etiology of IPF is unclear, but there is growing evidence that B cells and B cell receptor signaling play important roles in the pathogenesis of IPF. Zanubrutinib is a small molecule inhibitor of Bruton's tyrosine kinase (BTK), which is a key enzyme downstream of B cell receptor signaling pathway, has approved for the treatment of mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL). While its role in pulmonary fibrosis remains unknown. In this study, we explored the potential effect and mechanisms of zanubrutinib on pulmonary fibrosis in vivo and in vitro. METHODS: In the in vivo experiments, different doses of zanubrutinib were administered in a mouse model of bleomycin-induced pulmonary fibrosis, and pathological manifestations and lung function indices were evaluated. In vitro experiments were performed using TGF-ß1-stimulated fibroblasts to evaluate the effect of zanubrutinib on the activation and autophagy phenotype of fibroblasts and to explore the underlying signaling pathway mechanism. RESULTS: In vivo experiments demonstrated that zanubrutinib effectively attenuated bleomycin (BLM)-induced pulmonary fibrosis in mice. An in vitro mechanistic study indicated that zanubrutinib suppresses collagen deposition and myofibroblast activation by inhibiting the TGF-ß1/Smad pathway and induces autophagy through the TGF-ß1/mTOR pathway. CONCLUSIONS: Zanubrutinib alleviated bleomycin-induced lung fibrosis in mice by inhibiting the TGF-ß1 signaling pathway.

A decade of approved first-in-class small molecule orphan drugs: Achievements, challenges and perspectives.

In the past decade (2011-2020), there was a growing interest in the discovery and development of orphan drugs for the treatment of rare diseases. However, rare diseases only account for a population of 0.65‰-1‰ which usually occur with previously unknown biological mechanisms and lack of specific therapeutics, thus to increase the demands for the first-in-class (FIC) drugs with new biological targets or mechanisms. Considering the achievements in the past 10 years, a total of 410 drugs were approved by U.S. Food and Drug Administration (FDA), which contained 151 FIC drugs and 184 orphan drugs, contributing to make up significant numbers of the approvals. Notably, more than 50% of FIC drugs are developed as orphan drugs and some of them have already been milestones in drug development. In this review, we aim to discuss the FIC small molecules for the development of orphan drugs case by case and highlight the R&D strategy with novel targets and scientific breakthroughs.

Cabozantinib ameliorates lipopolysaccharide-induced lung inflammation and bleomycin--induced early pulmonary fibrosis in mice.

The lung, as the primary organ for gas exchange in mammals, is the main target organ for many pathogens and allergens, which may cause acute lung injury. A certain proportion of acute lung injury may progress into irreversible pulmonary fibrosis. Both acute lung injury and pulmonary fibrosis have high mortality rates and few effective treatments. Cabozantinib is a multi-target small molecule tyrosine kinase inhibitor and has been approved for the treatment of multiple malignant solid tumors. In this study, we explored the role of cabozantinib in acute lung injury and pulmonary fibrosis in vivo and in vitro. In the lipopolysaccharide and bleomycin induced mouse lung injury models, cabozantinib significantly improved the pathological state and reduced the infiltration of inflammatory cells in the lung tissues. In the bleomycin induced pulmonary fibrosis model, cabozantinib significantly reduced the area of pulmonary fibrosis and improved lung function in mice. The results of in vitro studies showed that cabozantinib could inhibit the inflammatory response and apoptosis of alveolar epithelial cells by inhibiting the activation of TLR4/NF-κB and NLRP3 inflammasome pathways. At the same time, cabozantinib could inhibit the activation of lung fibroblasts through suppressing the TGF-ß1/Smad pathway, and promote the apoptosis of fibroblasts. In summary, cabozantinib could alleviate lung injury through regulating the TLR4 /NF-κB/NLRP3 inflammasome pathway, and alleviate pulmonary fibrosis by inhibiting the TGF-ß1/Smad3 signaling pathway.

Synthesis and biological evaluation of resveratrol derivatives with anti-breast cancer activity.

Resveratrol is a natural phytoestrogen produced by plants to protect themselves from injury, UV irradiation, and fungal attack. The main active structure is E-resveratrol, which has many pharmacological activities. As the structure of resveratrol is similar to the natural estrogen 17ß-estradiol and the synthetic estrogen E-diethylstilbestrol, resveratrol is used in reducing the incidence of breast cancer. However, the therapeutic application of resveratrol is limited due to its low bioavailability. To improve its bioavailability and pharmacological activity, some resveratrol derivatives have been designed and synthesized by substitutions of methoxy, hydroxyl, and other functional groups or heterocyclic esterification either on the "A" or "B" ring, and double bonds were replaced by imine bonds and isometric heterocycles such as naphthyl and imidazole, or synthetic resveratrol oligomers. The structures, synthetic routes, and evaluation of the biological activities of these compounds are discussed. These are aimed at providing some references for the study of resveratrol derivatives in anti-breast cancer treatment.