Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Objective To explore the effective substances that specifically bind to RSV-F protein in the water extract of Forsythia suspensa,and verify its effect.Methods Surface plasmon resonance(SPR)affinity fishing technology was used to enrich,regenerate and recover the active components that specifically bind to RSV-F protein in the water extract of Forsythia suspensa.The recovered samples were analyzed and identified by UHPLC-MS/MS,and the active components that stably bind to F protein were screened by molecular docking,molecular dynamics simulation and SPR affinity detection,and then returned to animal experiments for pharmacodynamics verification.BALB/c mice were randomly divided into normal group,model group,positive control group,protocatechuic acid group and verbascoside group.In the experimental group,RSV was dripped into the nose to make models.Each drug group began to be given intragastric intervention on the day of modeling,and samples were taken after 4 days.HE staining was used to observe the pathological changes of lung tissue,and ELISA was used to detect the changes of inflammatory factors in lung tissue.Results A variety of active components in the aqueous extract of Forsythia suspensa could bind to RSV-F protein,among which the results of verbascoside and protocatechuic acid were more stable.In vivo pharmacodynamics experiments showed that the lung tissue of the model group had obvious pathological changes and the level of inflammatory factors increased,and the pathological state was improved after the intervention of verbascoside and protocatechuic acid compared with the normal group.Conclusion Forsythia suspensa can interfere with the expression of RSV-F protein through many active components such as verbascoside and protocatechuic acid,which may be the material basis of its anti-RSV.

This study presents prenatal diagnosis and genetic analysis of a pedigree with X-linked intellectual disability. A gravida at approximately 20 gestational weeks underwent prenatal diagnosis following non-invasive prenatal testing suggested sex chromosome abnormalities. Copy number variation (CNV) sequencing identified a 5.7 Mb duplication at Xp22.2p22.11 in the fetus, which initially classified as a variant of uncertain clinical significance. This duplication was inherited from the phenotypically normal mother, while paternal CNV results were normal. Genetic testing of four intellectually disabled family members revealed the identical 5.7 Mb duplication. Through expanded pedigree analysis, the pathogenicity classification of the Xp22.2p22.11 microduplication was upgraded to likely pathogenic. After comprehensive genetic counseling, the family elected pregnancy termination with informed consent.

Pancreatic ductal adenocarcinoma is profoundly treatment-resistant, due to intrinsic properties of the tumor cells and the complex tumor microenvironment. Consequently, surgical resection of the primary tumor is still the only intervention that significantly prolongs patient survival. This points at an urgent need for more effective (neo)adjuvant strategies to increase the fraction of patients eligible to surgery and to counter post-surgery disease recurrence. The advent of single-cell RNA-sequencing has created an opportunity for the development of a highly potent, patient-tailored adjuvant treatment that is based on the infusion of genetically engineered autologous T-cells armed with tumor-reactive T-cell receptors as identified in the patient′s own tumor sample.

Objective To explore the safety of Yttrium-90 resin microsphere selective internal radiation therapy (⁹⁰Y-SIRT) on malignant liver tumors. Methods A retrospective analysis was conducted on 64 patients with malignant liver tumors who underwent ⁹⁰Y-SIRT from February 2023 to November 2024 at Weifang People’s Hospital. The clinical characteristics of the patients and the occurrence of adverse reactions after treatment were analyzed to assess the safety of ⁹⁰Y-SIRT. Results Among the 64 patients, there were 52 males (81.25%) and 12 females (18.75%); the average age was (56.29±11.08) years. Seven patients (10.94%) had tumors with maximum diameter of less than 5 cm, 38 patients (59.38%) had tumors with maximum diameter of 5-10 cm, and 19 patients (29.68%) had tumors with maximum diameter of greater than 10 cm. There were 47 cases (73.44%) of solitary lesions and 17 cases (26.56%) of multiple lesions; 53 cases (82.81%) were primary liver cancers and 11 cases (17.19%) were metastatic liver cancers. Of the 64 patients, 63 successfully completed the Technetium-99m macroaggregated albumin (^99mTc-MAA) perfusion test and received the ⁹⁰Y-SIRT; one patient received ⁹⁰Y-SIRT after the second ^99mTc-MAA perfusion test due to a work error. The most common adverse reactions included grade 1 alanine aminotransferase (ALT) elevation in 26 cases (40.62%) and grade 2 in 2 cases (9.37%), grade 1 aspartate aminotransferase (AST) elevation in 27 cases (42.18%) and grade 2 in 7 cases (10.93%); grade 1 nausea in 17 cases (26.56%) and grade 2 in 6 cases (9.37%); grade 1 abdominal pain in 12 cases (18.75%), grade 2 in 5 cases (7.81%), and grade 3 in 1 case (1.56%); grade 1 vomiting in 11 cases (17.18%), grade 2 in 5 cases (7.81%), and grade 3 in 1 case (1.56%). Conclusion The adverse reactions of ⁹⁰Y-SIRT for treating malignant liver tumors are mild, indicating good safety.

In recent years, with the endless emergence of meibomian gland dysfunction(MGD)diagnostic equipment, rich treatment methods, and in-depth clinical and basic research on MGD at home and abroad, the understanding of MGD has entered a new stage. MGD-related dry eye is considered to be the main cause of lipid abnormal dry eye, and its occurrence and development is a chronic and multi-factorial pathological process. This article reviews the pathogenesis, imaging analysis and clinical treatment progress of MGD-related dry eye, in order to provide scientific evidence and ideas for clinical diagnosis and therapy of MGD-related dry eye.

BACKGROUND: Asthma is a common chronic inflammatory airway disease and intermittent hypoxia is increasingly recognized as a factor that may impact disease progression. The present study investigated whether intermittent hypoxia (IH) could aggravate asthma by promoting hypoxia-inducible factor-1α (HIF-1α)/nucleotide-binding oligomerization domain (NOD)-like receptor pyrin domain-containing protein 3 (NLRP3)/interleukin (IL)-1β-dependent pyroptosis and the inflammatory response and further elucidated the underlying molecular mechanisms involved. METHODS: A total of 49 patients diagnosed with severe bronchial asthma and diagnosed by polysomnography were enrolled at Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, between January 2022 and December 2022, and their general data and induced sputum were collected. BEAS-2B cells were treated with IL-13 and subjected to IH. An ovalbumin (OVA)-treated mouse model was also used to assess the effects of chronic intermittent hypoxia (CIH) on asthma. Pyroptosis, the inflammatory response, and related signalling pathways were assessed in vivo and in vitro . RESULTS: In this study, as the apnoea and hypopnea index (AHI) increased, the proportion of patients with uncontrolled asthma increased. The proportions of neutrophils and the levels of IL-6, IL-8, HIF-1α and NLRP3 in induced sputum were related to the AHI. NLRP3-mediated pyroptosis, which could be mediated by the HIF-1α signalling pathway, was activated in IL-13 plus IH-treated BEAS-2B cells and in the lungs of OVA/CIH mice. HIF-1α downregulation significantly reduced lung pyroptosis and ameliorated neutrophil inflammation by modulating the NLRP3/IL-1β pathway both in vitro and in vivo . Similarly, pretreatment with LW6, an inhibitor of HIF-1α, effectively blocked the generation of inflammatory cytokines in neutrophils. In addition, administration of the NLRP3 activator nigericin obviously increased lung neutrophil inflammation. CONCLUSIONS Obstructive sleep apnoea-hypopnea syndrome (OSAHS) is a risk factor for asthma exacerbation. IH aggravates neutrophil inflammation in asthma via NLRP3/IL-1β-dependent pyroptosis mediated by the HIF-1α signalling pathway, which should be considered a potential therapeutic target for the treatment of asthma with OSAHS.
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Humans ; Asthma/metabolism* ; Animals ; Pyroptosis/physiology* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Mice ; Signal Transduction/physiology* ; Male ; Hypoxia/metabolism* ; Female ; Interleukin-1beta/metabolism* ; Adult ; Inflammation/metabolism* ; Middle Aged ; Mice, Inbred C57BL

ObjectiveTo investigate the effects of foliar fertilizer of nano-calcium carbonate and calcium nitrate tetrahydrate on the agronomic traits， carbohydrate and endogenous hormone contents of Dendrobium officinale planted for 1 year under greenhouse cultivation， in order to provide scientific basis for fertilization to improve the yield and quality of D. officinale. MethodsSingle-factor experimental design was adopted. Starting from early spring， D. officinale was treated with foliar spraying according to corresponding fertilizers. Three treatment groups were established based on different fertilizers， namely， a blank group（clear water）， a nano-calcium carbonate group（0.727 g·L^-1 nano-calcium carbonate water-soluble fertilizer）， and a calcium nitrate tetrahydrate group（1.091 g·L^-1 calcium nitrate tetrahydrate water-soluble fertilizer）. The frequency of spraying was three times per month， and the entire treatment process lasted for nine months. The effects of various treatments on the traits and relative chlorophyll content of D. officinale were dynamically monitored. Sampling was conducted at three specific time points：August 2， 2023， September 8， 2023， and November 1， 2023， respectively. The contents of glucose and mannose in D. officinale stems were determined by high performance liquid chromatography（HPLC）， the content of soluble sugars in D. officinale stems and leaves was determined by phenol method， and enzyme-linked immunosorbent assay（ELISA） was used to detect the concentrations of cytokinin and auxin. ResultsCompared with the blank group， the treatments with nano-calcium carbonate and calcium nitrate tetrahydrate could significantly increase stem length， stem node number， leaf number， and tiller number. Among them， during the harvesting period in November， the stem length and tiller number， which are indicators related to the yield of D. officinale， increased by 60.85% and 19.23% after treatment with calcium nitrate tetrahydrate， and by 32.54% and 28.85% after treatment with nano-calcium carbonate， respectively. Compared with the blank group， treatments with nano-calcium carbonate and calcium nitrate tetrahydrate could promote the accumulation of sucrose in the stems and leaves of D. officinale to varying degrees， as well as the accumulation of polysaccharides， mannose， and glucose in the stems. In addition， nano-calcium carbonate treatment also facilitated the accumulation of fructose in the stems and leaves of D. officinale. Specifically， during the harvesting period in November， polysaccharides and mannose， which were the main active ingredients in D. officinale stems， increased by 28.48% and 29.36% after treatment with calcium nitrate tetrahydrate， and by 39.91% and 82.62% after treatment with nano-calcium carbonate， respectively. In addition， compared with the blank group， the concentrations of auxin in the stems and leaves of D. officinale were significantly increased after treatment with calcium nitrate tetrahydrate（P<0.05）. Similarly， the concentrations of cytokinin and auxin in the stems of D. officinale were also elevated after treatment with nano-calcium carbonate. Correlation analysis further indicated that elongation growth and tillering of D. officinale stems after foliar spraying of nano-calcium carbonate and calcium nitrate tetrahydrate might be related to the accumulation of carbohydrates in the stems and leaves and the synergistic effect of auxin and cytokinin. ConclusionIn production practice， spraying nano-calcium carbonate and calcium nitrate tetrahydrate can promote the accumulation of cytokinin， auxin， and carbohydrate contents in the stems and leaves of D. officinale， and promote tillering and elongation growth of the stems.

Objective To explore the effective substances that specifically bind to RSV-F protein in the water extract of Forsythia suspensa,and verify its effect.Methods Surface plasmon resonance(SPR)affinity fishing technology was used to enrich,regenerate and recover the active components that specifically bind to RSV-F protein in the water extract of Forsythia suspensa.The recovered samples were analyzed and identified by UHPLC-MS/MS,and the active components that stably bind to F protein were screened by molecular docking,molecular dynamics simulation and SPR affinity detection,and then returned to animal experiments for pharmacodynamics verification.BALB/c mice were randomly divided into normal group,model group,positive control group,protocatechuic acid group and verbascoside group.In the experimental group,RSV was dripped into the nose to make models.Each drug group began to be given intragastric intervention on the day of modeling,and samples were taken after 4 days.HE staining was used to observe the pathological changes of lung tissue,and ELISA was used to detect the changes of inflammatory factors in lung tissue.Results A variety of active components in the aqueous extract of Forsythia suspensa could bind to RSV-F protein,among which the results of verbascoside and protocatechuic acid were more stable.In vivo pharmacodynamics experiments showed that the lung tissue of the model group had obvious pathological changes and the level of inflammatory factors increased,and the pathological state was improved after the intervention of verbascoside and protocatechuic acid compared with the normal group.Conclusion Forsythia suspensa can interfere with the expression of RSV-F protein through many active components such as verbascoside and protocatechuic acid,which may be the material basis of its anti-RSV.

This study presents prenatal diagnosis and genetic analysis of a pedigree with X-linked intellectual disability. A gravida at approximately 20 gestational weeks underwent prenatal diagnosis following non-invasive prenatal testing suggested sex chromosome abnormalities. Copy number variation (CNV) sequencing identified a 5.7 Mb duplication at Xp22.2p22.11 in the fetus, which initially classified as a variant of uncertain clinical significance. This duplication was inherited from the phenotypically normal mother, while paternal CNV results were normal. Genetic testing of four intellectually disabled family members revealed the identical 5.7 Mb duplication. Through expanded pedigree analysis, the pathogenicity classification of the Xp22.2p22.11 microduplication was upgraded to likely pathogenic. After comprehensive genetic counseling, the family elected pregnancy termination with informed consent.