Fungal keratitis represents a significant cause of blindness, with current therapeutic approaches yielding limited success. The disease's onset and progression are primarily driven by fungal virulence factors and the host's immune response. The innate immune system is the first to respond, with neutrophils playing a pivotal role in the antifungal defense. Although neutrophils are critical for pathogen clearance, their excessive or abnormal activation can lead to tissue damage, exacerbating the disease. Thus, elucidating the mechanisms underlying neutrophil activity in fungal keratitis is crucial for refining treatment strategies. This article aims to systematically review the principal antimicrobial mechanisms employed by neutrophils, including phagocytosis, degranulation, and the formation of neutrophil extracellular traps(NETs). Furthermore, it explores the crosstalk between neutrophils and macrophages, alongside their collective impact and underlying mechanisms in the context of fungal keratitis. Exploration of the mechanisms of fungal keratitis facilitates precise intervention and enhances the efficacy of treatment.

ObjectiveTo explore the quantity-quality transfer process of medicinal materials， decoction pieces and standard decoction of Haliotidis Concha（Haliotis discus hannai） by analyzing the physical phase and compositional changes， so as to provide references for the effective control of its quality. MethodsA total of 20 batches of Haliotidis Concha（H. discus hannai） from different habitats were collected and prepared into corresponding calcined products and standard decoction， and the content of CaCO₃ of the three samples were determined and the extract yield and transfer rate of CaCO₃ were calculated. The changes in elemental composition and their relative contents were investigated by X-ray fluorescence spectrometry（XRF）， X-ray diffraction（XRD） was used to study the changes in the phase compositions of the three samples and to establish their respective XRD specific chromatogram. Fourier transform infrared spectrometry（FTIR） was used to study the changes in the chemical composition and content changes of the three samples and to establish their respective FTIR specific chromatogram， while combining hierarchical cluster analysis（HCA）， principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） to find the common and differential characteristics， in order to explore the quantity-quality transfer relationship in the preparation process of standard decoction of Haliotidis Concha（H. discus hannai）. ResultsThe CaCO₃ contents of the 20 batches of medicinal materials， decoction pieces and standard decoction of Haliotidis Concha（H. discus hannai） were 93.87%-98.95%， 96.02%-99.97% and 38.29%-51.96%， respectively， and the extract yield of standard decoction was 1.71%-2.37%， and the CaCO₃ transfer rate of decoction pieces-standard decoction was 0.68%-1.27%. XRF results showed that the elemental species and their relative contents contained in Haliotidis Concha and its calcined products had a high degree of similarity， and although there was no obvious difference in the elemental species contained in decoction pieces and standard decoction， the difference in the relative contents was obvious， which was mainly reflected in the decrease of the relative content of element Ca and the increase of the relative content of element Na. XRD results showed that Haliotidis Concha mainly contained CaCO₃ of aragonite and calcite， while calcined Haliotidis Concha only contained CaCO₃ of calcite， and standard decoction mainly contained CaCO₃ of calcite and Na₂CO₃ of natrite. FTIR results showed that there were internal vibrations of O-H， C-H， C=O， HCO₃^- and CO₃^2- groups in Haliotidis Concha， while O-H， HCO₃^- and CO₃^2- groups existed in the calcined products and standard decoction. ConclusionThe changes of Haliotidis Concha and calcined Haliotidis Concha are mainly the increase of CaCO₃ content， the transformation of CaCO₃ aragonite crystal form to calcite crystal form and the absence of organic components after calcination， and the changes of calcined products and standard decoction are mainly the decrease of CaCO₃ content and the increase of Na₂CO₃ relative content. The method established in the study is applicable to the quality control of the shellfish medicines-decoction pieces- standard decoction， which provides a new idea for the study of quality control of dispensing granules of shellfish medicines.

ObjectiveTo explore the quantity-quality transfer process of medicinal materials， decoction pieces and standard decoction of Haliotidis Concha（Haliotis discus hannai） by analyzing the physical phase and compositional changes， so as to provide references for the effective control of its quality. MethodsA total of 20 batches of Haliotidis Concha（H. discus hannai） from different habitats were collected and prepared into corresponding calcined products and standard decoction， and the content of CaCO₃ of the three samples were determined and the extract yield and transfer rate of CaCO₃ were calculated. The changes in elemental composition and their relative contents were investigated by X-ray fluorescence spectrometry（XRF）， X-ray diffraction（XRD） was used to study the changes in the phase compositions of the three samples and to establish their respective XRD specific chromatogram. Fourier transform infrared spectrometry（FTIR） was used to study the changes in the chemical composition and content changes of the three samples and to establish their respective FTIR specific chromatogram， while combining hierarchical cluster analysis（HCA）， principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） to find the common and differential characteristics， in order to explore the quantity-quality transfer relationship in the preparation process of standard decoction of Haliotidis Concha（H. discus hannai）. ResultsThe CaCO₃ contents of the 20 batches of medicinal materials， decoction pieces and standard decoction of Haliotidis Concha（H. discus hannai） were 93.87%-98.95%， 96.02%-99.97% and 38.29%-51.96%， respectively， and the extract yield of standard decoction was 1.71%-2.37%， and the CaCO₃ transfer rate of decoction pieces-standard decoction was 0.68%-1.27%. XRF results showed that the elemental species and their relative contents contained in Haliotidis Concha and its calcined products had a high degree of similarity， and although there was no obvious difference in the elemental species contained in decoction pieces and standard decoction， the difference in the relative contents was obvious， which was mainly reflected in the decrease of the relative content of element Ca and the increase of the relative content of element Na. XRD results showed that Haliotidis Concha mainly contained CaCO₃ of aragonite and calcite， while calcined Haliotidis Concha only contained CaCO₃ of calcite， and standard decoction mainly contained CaCO₃ of calcite and Na₂CO₃ of natrite. FTIR results showed that there were internal vibrations of O-H， C-H， C=O， HCO₃^- and CO₃^2- groups in Haliotidis Concha， while O-H， HCO₃^- and CO₃^2- groups existed in the calcined products and standard decoction. ConclusionThe changes of Haliotidis Concha and calcined Haliotidis Concha are mainly the increase of CaCO₃ content， the transformation of CaCO₃ aragonite crystal form to calcite crystal form and the absence of organic components after calcination， and the changes of calcined products and standard decoction are mainly the decrease of CaCO₃ content and the increase of Na₂CO₃ relative content. The method established in the study is applicable to the quality control of the shellfish medicines-decoction pieces- standard decoction， which provides a new idea for the study of quality control of dispensing granules of shellfish medicines.

The emergence of teeth is a complex physiological process characterized by the formation of the tooth crown, its movement towards the occlusal plane, and subsequent penetration through the alveolar bone and oral mucosa to achieve functional positioning for contact with opposing teeth. Parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHrP) are critical regulators of calcium and phosphorus metabolism in the body, playing significant roles in tooth emergence. Their regulatory functions exhibit intricate temporal and spatial dynamics, with underlying mechanisms that remain incompletely understood. In recent years, an increasing number of researchers both domestically and internationally have investigated the role and mechanisms of PTH/PTHrP in tooth emergence, primarily focusing on aspects such as dental sac formation, basal alveolar bone development, coronal alveolar bone resorption, root formation, and periodontal ligament development. Literature reviews indicate that PTH and PTHrP regulate bone metabolism, coordinate various signaling pathways including OPG/RANK/RANKL, cAMP/PKA, and Wnt/β-catenin, and are allosterically modulated by Ca2+ and ATP. These processes contribute to the development of dental sacs, which transmit signals to recruit osteoclasts and promote the resorption of crown alveolar bone, thereby forming an eruption pathway. Additionally, PTH/PTHrP plays a role in the formation of basal alveolar bone, root development, and the periodontal ligament, generating the force necessary for tooth eruption. Through precise spatiotemporal regulation and coordinated efforts, alveolar bone remodeling is achieved, facilitating the intricate process of tooth eruption. Through stringent temporal regulation and multi-faceted cooperation, remodeling of the alveolar bone occurs to complete this intricate developmental process of tooth emergence. Future research should further elucidate the mechanisms underlying PTH/PTHrP actions while also considering optimal dosage regimens regarding timing and frequency for therapeutic applications.

⁶⁰Co-γ ray irradiation has the unique advantages of high efficiency， strong penetration， operation at room temperature and no residue， which has been widely used in the sterilization of Chinese medicinal materials， decoction pieces， Chinese patent medicine. However， the irradiation effect may cause changes in the content of chemical components in Chinese materia medica or the emergence of new radiolysis products， leading to reduced efficacy and uncontrollable safety risks. This paper reviewed the relevant literature at home and abroad， summarized the effect of irradiation sterilization on various types of chemical compositions of Chinese medicinal materials and their preparations， and analyzed and explored the rule of change. The results showed that the content changes of various chemical components in Chinese materia medica after ⁶⁰Co-γ ray irradiation sterilization varied. The contents of most flavonoids， terpenoids， phenylpropanoids and quinones decreased after irradiation， and the degree of decrease increased with the elevated irradiation dose. The contents of lignans， alkaloids， isoflavones and some terpenoids did not change significantly before and after irradiation， while the content changes of triterpenoid saponins， dihydroflavonols， chalcones， sugars and glycosides after irradiation were not yet uniform. Therefore， it is recommended to pay attention to the compositional changes of irradiated Chinese medicines， strengthen the research on the standards of irradiated Chinese medicines， and standardize the irradiation and sterilization of Chinese medicines in order to promote the healthy and rational application of irradiated Chinese medicines.

ObjectiveTo explore whether miR-375 regulates the malignant characteristics of osteosarcoma (OS) by influencing the expression of MMP13. MethodsPlasmid DNAs and miRNAs were transfected into OS cells and HEK293 cells using Lipofectamine 3000 reagent. Real-time quantitative polymerase chain reaction was performed to measure the expression of miR-375 and MMP13 in OS patients and OS cells. Western blot was performed to analyze the MMP13 protein in the patients with OS and OS cells. The targeting relationship between miR-375 and MMP13 was analyzed by luciferase assay. Migration and invasion were analysed by heal wound and transwell assays, respectively. ResultsmiR-375 expression in OS tissues was lower than that in normal tissues. The expression of MMP13 was upregulated in OS tissues. MMP13 expression was negatively correlated withmiR-375 expression in patients with OS. Migration and invasion were significantly inhibited in OS cells with the miR-375 mimic compared with OS cells with the miRNA control. MMP13 partially reversed the inhibition of migration and invasion induced by miR-375 in the OS cells. ConclusionmiR-375 attenuates migration and invasion by downregulating the expression of MMP13 in OS cells.

Fermentum Rubrum（FR） is a kind of traditional Chinese medicine with dual-use functions of medicine and food， which has been used for more than 1 000 years. By referring to the ancient herbal classics and modern laws and regulations， the author sorted out the origin of FR， sorted out the development of processing， and analyzed the problems existing in the quality standard， aiming to provide a basis for further research and development of FR. There are many theories about the origin of FR. After summarizing and sorting out the relevant literature， three viewpoints are mainly drawn， including Han dynasty origin theory， Wei and Jin origin theory and Tang dynasty origin theory. After synthesizing the views of various schools， it is believed that the origin of FR should be no later than the Eastern Han dynasty. FR is made from rice by fermentation， which has the effect of strengthening the spleen， eliminating food， promoting blood circulation and removing stasis after fermentation. Although the natural fermentation in ancient times has been able to pay attention to the influence of temperature， humidity and strain on the quality of FR， due to the low level of science and technology， there are still problems such as complicated and cumbersome process， large workload and high labor cost. To the pure fermentation in modern times， the fermentation processing method of FR has been evolved， gradually improved and perfected. However， due to the lack of standardized research， there is no unified standard for the fermentation process of FR in various regions， and the quality standard lags behind seriously， which leads to the unstable product quality on the market. Among the 15 specifications for the preparation of FR， only 6 have been published in the past 5 years， and most of them have not been revised in a timely manner， and some of them have not been included in the updated version. Based on this， it is recommended to carry out a systematic study on processing technology of FR， and the best process is selected to accelerate the revision and improvement of its standardization， so as to improve the quality of FR sold in the market and promote its stable and sustainable development.

【Objective】 To analyze the effects of different fortified feeding methods on nutritional metabolism and growth rate of preterm very low birth weight infants (VLBWI), in order to provide new clues for improving the prognosis of the preterm infants. 【Methods】 A total of 115 cases of premature VLBWI admitted to Department of Neonatology, The First Affiliated Hospital of Kunming Medical University from January 2019 to December 2020 were included in this study, and were divided into fortified breastfeeding group (HFM group), mixed feeding group, and premature formula feeding group (PF group) based on their feeding methods. The effects of different feeding methods on the nutritional metabolism and growth rate of premature VLBWI were analyzed. 【Results】 1) The hospitalization time of infants in the HFM group was shorter than that in PF group and mixed feeding group (t=7.185, 6.924, P<0.05). 2) The proportion of necrotizing enterocolitis (NEC) in the HFM group during hospitalization was lower than that in the PF group (P<0.05); the proportions of late onset septicemia(LOS) and extra uterine growth restriction(EUGR) in the HFM group during hospitalization were lower than those in the PF group (χ²=5.030, 4.147, P<0.05); the proportion of LOS was lower than that of the mixed feeding group(χ²=6.589, P<0.05). 3) During hospitalization, the proportions of abdominal distension, bloody stools and increased eosinophils in the HFM group were lower than those in the PF group (P<0.05), which in mixed feeding group was lower than those in PF group (Fisher exact test, P<0.05). 4) At discharge, the weight and length growth rate of the HFM group were higher than those of the mixed feeding group (t=3.722, 0.425, P<0.001) and the PF group (t =6.015, 0.496, P< 0.001). 【Conclusion】 Fortified breastfeeding can more effectively increase the growth rate of VLBWI in premature infants, improve nutritional metabolism, reduce complications and adverse feeding reactions related to premature infants, and is safer and more effective.

To construct a recombinant expression system for a single-domain antibody targeting the urease of Helicobacter pylori(Hp),this study employed several strategies.First,using artificial intelligence(AI)auxiliary tools such as Pymol,I-TASSER,and ClussPro2,the molecular interactions between different antibodies and Hp urease subunit B(UreB)were analyzed.The fully humanized single-domain antibody UreBAb was identified as the primary research target.Next,the UreBAb gene sequence was optimized based on Escherichia coli codon preferences,and was inserted into expression vectors such as pET28a and pE-SUMO.The resulting recombinant expression strains were obtained by transforming Escherichia coli Rosetta(DE3).Recombinant antibody proteins were prepared through IPTG induction,and its activity was detected using extracted Hp urease as the antigen.SDS-PAGE analysis confirmed the correct expression of both UreBAb and SUMO-UreBAb,with protein yields of 0.34 mg/mL and 0.41 mg/mL,respectively.Unidirectional immunodiffusion experiments further confirmed that both recombinant antibodies exhibited strong affinity for Hp UreB antigen,with inhibition rates of 51.27%and 74.07%,respectively.Additionally,leveraging artificial intelligence tools such as AlphaFold2,cluspro2,mCSM-AB,OSPREY,and FoldX,the study evaluated and analyzed key binding sites and mutational strategies affecting the stability of the antigen-antibody complex.Subsequently,nine UreBAb evolution mutants were constructed,and their binding activities with the antigen were enhanced.Among these,the I107W mutant showed the most significant improvement,achieving a 24.95%increase compared to the wild-type UreBAb.This research lays a solid foundation for the development of fully humanized single-domain antibodies against Hp.

Objective:To compare the effect of anesthesia mode on the neurological functional outcomes in patients undergoing endovascular treatment for acute posterior circulation ischemic stroke.Methods:Clinical data of 656 patients undergoing intravascular therapy for acute posterior circulation ischemic stroke registered in online Acute Stroke Patients for Stroke Management Quality Evaluation Database from January 2017 to December 2022 were retrospectively analyzed.The data included 163 cases with conscious sedation and 493 cases with general anesthesia during the procedure.After propensity score matching,428 patients were included in the analysis,including 155 cases in the conscious sedation group and 273 cases in the general anesthesia group.The differences of operation mode,etiology type,vascular recanalization,hemorrhagic transformation at 24 h,modified Rankin Scale(mRS)score at 3 months and mortality within 3 months were compared between the two groups.Binary logistic regression was used to explore the effect of different anesthesia mode on neurological functional outcomes.Results:There was a significant difference in operation mode between the two groups(P<0.01),while there were no significant differences in etiology type,vascular recanalization,hemorrhagic transformation at 24 h,mRS score at 3 months or mortality within 3 months(all P>0.05).Binary logistic regression analysis revealed that anesthesia modes were not significantly associated with functional outcomes of patients(OR=1.151,95%CI:0.751-1.765,P>0.05).Conclusion:Anesthesia mode(conscious sedation or general anesthesia)will not affect the neurological functional outcomes in patients with acute posterior circulation ischemic stroke undergoing endovascular treatment.