Extracellular vesicles(EVs)are nanoscale membrane vesicles released by cells that carry bioactive molecules such as proteins and nucleic acids. They play a key role in intercellular communication. In recent years, they have shown significant potential in the diagnosis and treatment of corneal and ocular surface diseases. As diagnostic biomarkers, EVs derived from tears or other sources can reflect disease-specific characteristics, providing a basis for the early disease detection. In terms of therapy, mesenchymal stem cells(MSCs)-derived EVs deliver miRNAs to modulate inflammatory pathways, promote corneal epithelial repair, and inhibit fibrosis. Furthermore, as natural drug vehicles, EVs enable precise delivery of therapeutic agents and promote corneal wound healing. This review systematically summarizes the recent research advances in the application of EVs in the field.

Abnormal blinking can manifest in various ways, including a significant increase or decrease in blink frequency, incomplete blinking, and uncoordinated blinking movements. Currently, a unified diagnostic criterion is still lacking. Abnormal blinking is closely linked to factors such as ocular surface diseases, neurodevelopmental disorders, and visual function abnormalities. In recent years, an increasing number of pediatric patients have visited ophthalmic clinics with abnormal blinking as their primary complaint. With the rising incidence of pediatric meibomian gland dysfunction(MGD)and dry eye disease, their proportion among cases of abnormal blinking has also grown continuously.This article reviews the research progress on meibomian gland changes in children with abnormal blinking. It aims to provide guidance for the early prevention and clinical care of pediatric MGD, reduce the incidence of MGD, and shorten the treatment cycle, thereby providing a solid basis for clinical diagnosis, treatment, and precise prevention.

In-depth study of the components of polymyxins is the key to controlling the quality of this class of antibiotics. Similarities and variations of components present significant analytical challenges. A two-dimensional (2D) liquid chromatography-mass spectrometr (LC-MS) method was established for screening and comprehensive profiling of compositions of the antibiotic colistimethate sodium (CMS). A high concentration of phosphate buffer mobile phase was used in the first-dimensional LC system to get the components well separated. For efficient and high-accuracy screening of CMS, a targeted method based on a self-constructed high resolution (HR) mass spectrum database of CMS components was established. The database was built based on the commercial MassHunter Personal Compound Database and Library (PCDL) software and its accuracy of the compound matching result was verified with six known components before being applied to genuine sample screening. On this basis, the unknown peaks in the CMS chromatograms were deduced and assigned. The molecular formula, group composition, and origins of a total of 99 compounds, of which the combined area percentage accounted for more than 95% of CMS components, were deduced by this 2D-LC-MS method combined with the MassHunter PCDL. This profiling method was highly efficient and could distinguish hundreds of components within 3 h, providing reliable results for quality control of this kind of complex drugs.

To address the challenges in blood cell recognition caused by diverse morphology, dense distribution, and the abundance of small target information, this paper proposes a blood cell detection algorithm - the "You Only Look Once" model based on hybrid mixing attention and deep over-parameters (HADO-YOLO). First, a hybrid attention mechanism is introduced into the backbone network to enhance the model's sensitivity to detailed features. Second, the standard convolution layers with downsampling in the neck network are replaced with deep over-parameterized convolutions to expand the receptive field and improve feature representation. Finally, the detection head is decoupled to enhance the model's robustness for detecting abnormal cells. Experimental results on the Blood Cell Counting Dataset (BCCD) demonstrate that the HADO-YOLO algorithm achieves a mean average precision of 90.2% and a precision of 93.8%, outperforming the baseline YOLO model. Compared with existing blood cell detection methods, the proposed algorithm achieves state-of-the-art detection performance. In conclusion, HADO-YOLO offers a more efficient and accurate solution for identifying various types of blood cells, providing valuable technical support for future clinical diagnostic applications.
Algorithms ; Humans ; Blood Cells/cytology* ; Blood Cell Count/methods* ; Neural Networks, Computer ; Deep Learning ; Detection Algorithms

Objective:To investigate the effect of Huanshaodan on improving learning and memory impairment in a mouse model of Alzheimer's disease (AD) which was named with senescence accelerated mouse-prone 8(SAMP8), as well as the neuroinflammatory response mechanisms mediated by the p38 mitogen activated protein kinase (p38MAPK) and extracellular signal regulated protein kinase 1/2 (ERK1/2) signaling pathways.Methods:Seven-month-old SPF grade male SAMP8 mice were randomly assigned into three groups(6 mice in each group) using a random number table: model group, low-dose Huanshaodan group(1.17g/kg, twice daily via gavage), and high-dose Huanshaodan group(2.34g/kg, twice daily via gavage).Weight-matched seven-month-old male mice with anti-rapid aging traits(senescence-accelerated mouse-resistant 1, SAMR1)were designated as the normal control group( n=6).The mice in control group and the model group received 0.9% NaCl via gavage twice daily.All mice underwent continuous interventions for 28 days.The learning and memory abilities were assessed by Morris water maze.Immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein(GFAP) and ionized calcium-binding adaptor molecule-1(Iba1) as markers for astrocytes and microglial cells in the hippocampus, respectively.ELISA was used to measure pro-inflammatory cytokines interleukin-6(IL-6), interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in hippocampal tissue.Western blot was used for analyzing the expression levels of pro-inflammatory enzymes, inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2), as well as phosphorylated levels of ERK1/2 and p38MAPK in hippocampal tissue.Data were statistically analyzed using SPSS 20.0.The repeated measures analysis of variance or one-way analysis of variance was used for multi groups comparison. Results:Morris water maze test results indicated interactions between time and group in the escape latencies of the four groups of mice( F=3.787, P<0.05).From the 5th to 6th day, the escape latencies of the low- and high-dose Huanshaodan groups were lower than those of the model group(both P<0.05).On the 4th to 6th day, the escape latencies of the high-dose Huanshaodan group were lower than those of the low-dose group(all P<0.05).Significant differences were observed in the residence time in the target quadrant and the number of crossing the platform among the four groups of mice( F=8.587, 12.633, both P<0.05).The residence time in the target quadrant of the model group((17.8±3.4)s) and the number of crossing the platform((1.6±0.6)times)were less than those of the control group((40.6±3.7)s, (4.6±0.6)times) and high-dose Huanshaodan group(31.8±4.0)s, (2.8±0.8)times), all P<0.05).Western blot results indicated significant differences in the expression of iNOS, COX-2, p-p38MAPK/p38MAPK, and p-ERK1/2/ERK1/2 in the hippocampal tissues of the four groups of mice( F=207.516, 10.627, 108.497, 34.330, all P<0.05) and the indexes in model group were all higher than those of control group and high-dose Huanshaodan group(all P<0.05).ELISA results revealed significant differences in the levels of IL-6, TNF-α and IL-1β in the serum of the four groups of mice( F=66.790, 82.424, 42.919, all P<0.05), and the indexes of model group were higher than those of the other three groups(all P<0.05).Immunofluorescence results showed significant differences in the relative fluorescence intensity of GFAP and Iba1 in the hippocampal tissues of the four groups of mice( F=20.269, 56.437, both P<0.05).The relative fluorescence intensity values of GFAP and Iba1 in the hippocampal tissues of the high-dose Huanshaodan group were lower than those of the model group(both P<0.05), while the expression level of Iba1 in high-dose group was lower than that in the low-dose group( P<0.05). Conclusion:High-dose Haunshaodan may inhibit the activation of hippocampal glial cells by blocking the p38MAPK and ERK1/2 pathways, reducing neuroinflammation, then improving learning and memory disorders in SAMP8 mice.

Objective To study the effect of Liuwei Dihuangwan on autophagy level and its mechanism in SAMP8 mice and Aβ-stimulated BV2 cell model,and to explore the molecular mechanism of tonifying the kidney and filling up the essence to prevent and control Alzheimer's disease(AD)through interfering with autophagy.Methods Ten 7-month-old male anti-aging mice(SAMR1)were taken as the normal group,and 40 7-month-old male rapid aging mice(SAMP8)were randomly control and model groups,equal volumes of saline were administered by gavage twice a day for 4 weeks,and the levels of Aβ expression in the hippocampus of the mice in each group were detected by immunofluorescence;The expression levels of FcγRⅡB,c-Src and SHP-1 in the hippocampus of each group were detected by Western blot;BV2 cells were cultured and Fcγ receptor Ⅱ-b(FcγRⅡB)overexpression vectors were constructed;the AD state cell model was established by treating the BV2 cells with 5 μmol·L-1 Aβ1-42,and the Liuwei Dihuangwan drug-containing serum was prepared.The cells were divided into NC group,Aβ1-42 group,blank serum group,drug-containing serum group,Vector group,FcγRⅡB OE group,and drug-containing serum+FcγRⅡB OE group;immunofluorescence was used to detect the expression level of Aβ protein in the cells of each group;Western blot was used to detect the expression level of p62,LC3 Ⅱ/Ⅰ,FcγRⅡB,SHP-1,and c-Src in cells of each group.Results Compared with the normal group,the hippocampal Aβ,FcγRⅡB,SHP-1,and c-Src expression levels in the model group of mice were significantly higher(P<0.01),and compared with the model group,the expression levels of Aβ,FcγRⅡB,SHP-1,and c-Src in the low-,medium-,and high-dose groups of Liuwei Dihuangwan were significantly lower(P<0.01),it also showed a significant dose dependent relationship.Compared with NC group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in Aβ1-42 group were significantly increased(P<0.01),and LC3Ⅱ/Ⅰ was significantly decreased(P<0.01);Compared with Aβ1-42 group and blank serum group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in drug-containing serum group were significantly decreased(P<0.01),and LC3Ⅱ/Ⅰ was significantly increased(P<0.01);Compared with NC group and Vector group,the expression of Aβ in FcγRⅡB OE group was increased,the protein expressions of p62,FcγRⅡB,SHP-1 and c-Src were significantly increased(P<0.01),and LC3Ⅱ/Ⅰ was significantly decreased(P<0.01);Compared with the drug-containing serum group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in the drug-containing serum+FcγRⅡB OE group were significantly increased(P<0.01),and the protein expression levels of LC3Ⅱ/Ⅰ were significantly decreased(P<0.01).Conclusion Liuwei Dihuangwan improved AD by inhibiting microglia FcγRⅡB/c-Src pathway and increasing autophagy level.

Objective:To investigate the effect of Huanshaodan on improving learning and memory impairment in a mouse model of Alzheimer's disease (AD) which was named with senescence accelerated mouse-prone 8(SAMP8), as well as the neuroinflammatory response mechanisms mediated by the p38 mitogen activated protein kinase (p38MAPK) and extracellular signal regulated protein kinase 1/2 (ERK1/2) signaling pathways.Methods:Seven-month-old SPF grade male SAMP8 mice were randomly assigned into three groups(6 mice in each group) using a random number table: model group, low-dose Huanshaodan group(1.17g/kg, twice daily via gavage), and high-dose Huanshaodan group(2.34g/kg, twice daily via gavage).Weight-matched seven-month-old male mice with anti-rapid aging traits(senescence-accelerated mouse-resistant 1, SAMR1)were designated as the normal control group( n=6).The mice in control group and the model group received 0.9% NaCl via gavage twice daily.All mice underwent continuous interventions for 28 days.The learning and memory abilities were assessed by Morris water maze.Immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein(GFAP) and ionized calcium-binding adaptor molecule-1(Iba1) as markers for astrocytes and microglial cells in the hippocampus, respectively.ELISA was used to measure pro-inflammatory cytokines interleukin-6(IL-6), interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in hippocampal tissue.Western blot was used for analyzing the expression levels of pro-inflammatory enzymes, inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2), as well as phosphorylated levels of ERK1/2 and p38MAPK in hippocampal tissue.Data were statistically analyzed using SPSS 20.0.The repeated measures analysis of variance or one-way analysis of variance was used for multi groups comparison. Results:Morris water maze test results indicated interactions between time and group in the escape latencies of the four groups of mice( F=3.787, P<0.05).From the 5th to 6th day, the escape latencies of the low- and high-dose Huanshaodan groups were lower than those of the model group(both P<0.05).On the 4th to 6th day, the escape latencies of the high-dose Huanshaodan group were lower than those of the low-dose group(all P<0.05).Significant differences were observed in the residence time in the target quadrant and the number of crossing the platform among the four groups of mice( F=8.587, 12.633, both P<0.05).The residence time in the target quadrant of the model group((17.8±3.4)s) and the number of crossing the platform((1.6±0.6)times)were less than those of the control group((40.6±3.7)s, (4.6±0.6)times) and high-dose Huanshaodan group(31.8±4.0)s, (2.8±0.8)times), all P<0.05).Western blot results indicated significant differences in the expression of iNOS, COX-2, p-p38MAPK/p38MAPK, and p-ERK1/2/ERK1/2 in the hippocampal tissues of the four groups of mice( F=207.516, 10.627, 108.497, 34.330, all P<0.05) and the indexes in model group were all higher than those of control group and high-dose Huanshaodan group(all P<0.05).ELISA results revealed significant differences in the levels of IL-6, TNF-α and IL-1β in the serum of the four groups of mice( F=66.790, 82.424, 42.919, all P<0.05), and the indexes of model group were higher than those of the other three groups(all P<0.05).Immunofluorescence results showed significant differences in the relative fluorescence intensity of GFAP and Iba1 in the hippocampal tissues of the four groups of mice( F=20.269, 56.437, both P<0.05).The relative fluorescence intensity values of GFAP and Iba1 in the hippocampal tissues of the high-dose Huanshaodan group were lower than those of the model group(both P<0.05), while the expression level of Iba1 in high-dose group was lower than that in the low-dose group( P<0.05). Conclusion:High-dose Haunshaodan may inhibit the activation of hippocampal glial cells by blocking the p38MAPK and ERK1/2 pathways, reducing neuroinflammation, then improving learning and memory disorders in SAMP8 mice.

Objective To study the effect of Liuwei Dihuangwan on autophagy level and its mechanism in SAMP8 mice and Aβ-stimulated BV2 cell model,and to explore the molecular mechanism of tonifying the kidney and filling up the essence to prevent and control Alzheimer's disease(AD)through interfering with autophagy.Methods Ten 7-month-old male anti-aging mice(SAMR1)were taken as the normal group,and 40 7-month-old male rapid aging mice(SAMP8)were randomly control and model groups,equal volumes of saline were administered by gavage twice a day for 4 weeks,and the levels of Aβ expression in the hippocampus of the mice in each group were detected by immunofluorescence;The expression levels of FcγRⅡB,c-Src and SHP-1 in the hippocampus of each group were detected by Western blot;BV2 cells were cultured and Fcγ receptor Ⅱ-b(FcγRⅡB)overexpression vectors were constructed;the AD state cell model was established by treating the BV2 cells with 5 μmol·L-1 Aβ1-42,and the Liuwei Dihuangwan drug-containing serum was prepared.The cells were divided into NC group,Aβ1-42 group,blank serum group,drug-containing serum group,Vector group,FcγRⅡB OE group,and drug-containing serum+FcγRⅡB OE group;immunofluorescence was used to detect the expression level of Aβ protein in the cells of each group;Western blot was used to detect the expression level of p62,LC3 Ⅱ/Ⅰ,FcγRⅡB,SHP-1,and c-Src in cells of each group.Results Compared with the normal group,the hippocampal Aβ,FcγRⅡB,SHP-1,and c-Src expression levels in the model group of mice were significantly higher(P<0.01),and compared with the model group,the expression levels of Aβ,FcγRⅡB,SHP-1,and c-Src in the low-,medium-,and high-dose groups of Liuwei Dihuangwan were significantly lower(P<0.01),it also showed a significant dose dependent relationship.Compared with NC group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in Aβ1-42 group were significantly increased(P<0.01),and LC3Ⅱ/Ⅰ was significantly decreased(P<0.01);Compared with Aβ1-42 group and blank serum group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in drug-containing serum group were significantly decreased(P<0.01),and LC3Ⅱ/Ⅰ was significantly increased(P<0.01);Compared with NC group and Vector group,the expression of Aβ in FcγRⅡB OE group was increased,the protein expressions of p62,FcγRⅡB,SHP-1 and c-Src were significantly increased(P<0.01),and LC3Ⅱ/Ⅰ was significantly decreased(P<0.01);Compared with the drug-containing serum group,the protein expressions of Aβ,p62,FcγRⅡB,SHP-1 and c-Src in the drug-containing serum+FcγRⅡB OE group were significantly increased(P<0.01),and the protein expression levels of LC3Ⅱ/Ⅰ were significantly decreased(P<0.01).Conclusion Liuwei Dihuangwan improved AD by inhibiting microglia FcγRⅡB/c-Src pathway and increasing autophagy level.

OBJECTIVE To test and compare the median effective dose(ED50) of ciprofol for gastroscope in patients of different genders and ages. METHODS Patients who planed to undergo gastroscope examination and treatment from March 2023 to April 2023 were selected, and divided into four groups according to stratified random method: N1 group(non-elderly male patients), N2 group(non-elderly female patients), N3 group(elderly male patients), and N4 group(elderly female patients). All patients received intravenous injection of 0.1 μg·kg−1 sufentanil followed by injection of the test dose of ciprofol according to Dixon’s modified sequential method. Gastroscope was performed after the disappearance of the eyelash reflex. The initial dose of ciprofol in all four groups was 0.4 mg·kg−1, and the ratio of adjacent doses was 1∶1.1. The next patient would receive a 10% increase in the dose of ciprofol if the patient experienced positive reactions such as coughing, frowning, and body movements during the endoscopy process. Otherwise, it would be judged as a negative reaction, and the next patient would receive a 10% decrease in the dose of ciprofol. The transition from a positive reaction to a negative reaction was defined as a turning point, and the study was terminated when seven turning points occurred. Hemodynamic parameters, oxygen saturation and adverse reactions were recorded at different time points. The Probit regression analysis method was used to calculate the ED50 of ciprofol for four groups. RESULTS The ED50 of ciprofol combined with 0.1 μg·kg−1 sufentanil for gastroscope in the non-elderly men, non-elderly women, elderly men, and elderly women were 0.409, 0.373, 0.356, 0.327 mg·kg−1, respectively. The ED50 of ciprofol in the N1 group was significantly higher compared with the N2 group and N3 group(P<0.05). The ED50 of ciprofol in the N4 group was significantly lower compared with the N2 group and N3 group(P<0.05). CONCLUSION The ED50 of ciprofol is significantly different among gastroscope patients of different genders and ages, which is lower in female patients than in male patients, and is lower in older patients than in non-elderly patients.