ObjectiveTo investigate the effect of Yaoshu Zhuyu Fang on the regulation of the microRNA-17-5P (miR-17-5P)/murine double minute 2 (MDM2)/p53 axis in the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells, and its potential molecular mechanism. MethodsIntervertebral disc annulus fibrosus tissues were obtained from 8-week-old SPF-grade male SD rats, and annulus fibrosus cells were isolated and obtained by enzyme digestion and mechanical dispersion. Annulus fibrosus cells were divided into 6 groups: Group C was the blank control group, in which annulus fibrosus cells were not treated with interleukin-1β (IL-1β) but were cultured in RPMI 1640 complete medium. Group β was the degeneration model group constructed by treating annulus fibrosus cells with 10 ng/mL IL-1β for 24 h. Group β+B was the IL-1β + blank serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% blank serum for 24 h. Group β+W was the IL-1β + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. Group β+I was the IL-1β + miR-17-5P inhibitor group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor. Group β+I+W was the IL-1β + miR-17-5P inhibitor + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor, and finally treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. CCK-8 assay was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis. Real-time quantitative PCR was used to detect the expression levels of miR-17-5P, MDM2 mRNA, and p53 mRNA in cells. Western blotting was used to detect the protein expression levels of MDM2 and p53 in cells. Dual-luciferase reporter system was used to analyze the targeting relationship between miR-17-5P and MDM2. ResultsCompared with Group C, Group β showed a significant decrease in cell survival rate (P<0.001), a significant increase in cell apoptosis rate (P<0.001), significantly increased expression of miR-17-5P, p53 mRNA, and p53 protein (P<0.001), and significantly decreased expression of MDM2 mRNA and protein (P<0.001). Compared with Group β, Group β+W, Group β+I, and Group β+I+W showed significantly increased cell survival rate, significantly decreased apoptosis rate, significantly decreased expression of miR-17-5P, p53 mRNA, and p53 protein, and significantly increased expression of MDM2 mRNA and protein (P<0.001). Moreover, changes in the above indicators were greater in Group β+I+W (P<0.001). Circular RNA Interactome predicted that miR-17-5P had specific binding sites with the 3' untranslated region (3'UTR) of MDM2. Transfection of miR-17-5P mimic significantly reduced the luciferase expression level of co-transfected luciferase reporter plasmid containing wild-type MDM2 3'UTR (P<0.05), but had no significant effect on luciferase expression in cells co-transfected with luciferase reporter plasmid containing mutant MDM2 3'UTR (P>0.05). ConclusionYaoshu Zhuyu Fang down-regulates the expression of miR-17-5P, promotes the synthesis of MDM2 protein, thereby down-regulates p53, promotes proliferation, and inhibits the apoptosis of rat intervertebral disc annulus fibrosus cells.

The accurate and timely detection of biochemical coagulation indicators is pivotal in pulmonary and critical care medicine. Despite their reliability, traditional laboratories often lag in terms of rapid diagnosis. Point-of-care testing (POCT) has emerged as a promising alternative, which is awaiting rigorous validation. We assessed 226 samples from patients at the First Affiliated Hospital of Guangzhou Medical University using a Beckman Coulter AU5821 and a PUSHKANG POCT Biochemistry Analyzer MS100. Furthermore, 350 samples were evaluated with a Stago coagulation analyzer STAR MAX and a PUSHKANG POCT Coagulation Analyzer MC100. Metrics included thirteen biochemical indexes, such as albumin, and five coagulation indices, such as prothrombin time. Comparisons were drawn against the PUSHKANG POCT analyzer. Bland-Altman plots (MS100: 0.8206‒0.9995; MC100: 0.8318‒0.9911) evinced significant consistency between methodologies. Spearman correlation pinpointed a potent linear association between conventional devices and the PUSHKANG POCT analyzer, further underscored by a robust correlation coefficient (MS100: 0.713‒0.949; MC100: 0.593‒0.950). The PUSHKANG POCT was validated as a dependable tool for serum and whole blood biochemical and coagulation diagnostics. This emphasizes its prospective clinical efficacy, offering clinicians a swift diagnostic tool and heralding a new era of enhanced patient care outcomes.
Humans ; Point-of-Care Testing ; Critical Care ; Blood Coagulation Tests/methods* ; Male ; Blood Coagulation ; Female ; Middle Aged ; Reproducibility of Results ; Prothrombin Time ; Aged ; Adult ; Point-of-Care Systems

The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines. Therefore, there remains a need for alternative antivirals that target processes less susceptible to mutations, such as the formation of six-helix bundle (6-HB) during the viral fusion step of host cell entry. In this study, a novel high-throughput screening (HTS) assay employing a yeast-two-hybrid (Y2H) system was established to identify inhibitors of HR1/HR2 interaction. The compound IMB-9C, which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity, was selected. IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion. It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex. In addition, virtual docking and site-mutagenesis results suggest that amino acid residues A930, I931, K933, T941, and L945 are critical for IMB-9C binding to HR1. Collectively, in this study, we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.

Objective:To explore the application value of arthroscopic tunnel pulling and suturing methods in West type Ⅱ and Ⅲ lateral meniscus posterior root injuries.Methods:A total of 184 patients with West Ⅱ ( n=92) and Ⅲ ( n=92) lateral meniscus posterior root injuries who underwent surgical treatment in Second Affiliated Hospital of Xi′an Jiaotong University from May 2022 to June 2023 were selected as the study subjects, by adopting a retrospective controlled study method. Using a random number table method, West Ⅱ and Ⅲ patients were divided into a tunnel method group and a suture method group, with 46 patients in each group. There were 23 male and 23 female patients in the West Ⅱ tunnel method group, with an age of (48.36±2.52) years. There were 25 male and 21 female patients in the suture method group, with an age of (48.61±2.21) years. There were 24 male and 22 female patients in the West Ⅲ tunnel method group, with an age of (48.53±2.36) years. There were 24 male and 22 female patients in the suture method group, with an age of (46.38±2.67) years. The tunnel method group used arthroscopic tibial tunnel cable fixation method, while the suture method group used arthroscopic full internal meniscus suture method. Compared the surgical time, length of hospital stay, and first postoperative time of getting out of bed between the tunnel method group and the suture method group of patients with different subtypes. Compared the knee joint function scores (Lysholm, Tegner, IKDC, VAS) before and after surgery. Used MRI to examine the compression of the patient′s lateral meniscus. Compared the incidence of adverse reactions. Measurement data with normal distribution were represented as mean ± standard deviation( ± s), and the comparison between groups was conducted using the t-test; the comparison of count data were represented as [ n(%)] and was conducted by chi-square test or Fisher exact probability. The skewness data were expressed by M( Q1, Q3), and rank-sum test was used for inter-group comparison. Results:There was no significant difference in surgical time, length of hospital stay, and first postoperative bed time between the West type Ⅱ, Ⅲ tunel method group and the suture method group ( P>0.05). At 12 months after surgery, there was no statistically significant difference in the Lysholm score, Tegner score, IKDC score, and VAS score between the West Ⅱ type patients in the tunnel method group and the suture method group ( P>0.05). However, the Lysholm score and IKDC score in the West Ⅲ type tunnel method group were significantly higher than those in the suture method group, and the difference was statistically significant( P<0.05). However, there was no statistically significant difference in the Tegner score and VAS score ( P>0.05). At the 12th months after surgery, there was no statistically significant difference in the overall width of the meniscus between the tunnel method group and the suture method group for patients with different West classifications( P>0.05). However, the relative percentage of meniscus compression in the tunnel method group was significantly lower than that in the suture method group, and the difference was statistically significant ( P<0.05). There was no statistically significant difference in the incidence of postoperative complications between the tunnel method group and the suture method group in patients with different West subtypes ( P>0.05). Conclusions:The arthroscopic tibial tunnel wire fixation method is effective, safe, and reliable in treating patients with West Ⅱ and West Ⅲ lateral meniscus posterior root injuries promoting kneejoint function recovery. It is expected to become a high-quality treatment plan for patients with West Ⅱ and West Ⅲ meniscus injuries.

Objective To explore the methylation status of EGFR promoter region in peripheral blood of EGFR driver gene-positive lung adenocarcinoma patients and its clinical significance.Methods The methylation of EGFR promoter region in peripheral blood DNA of 32 EGFR driver gene-positive lung adenocarcinoma patients and 24healthy controls were detected by(matrix-assisted laser desorp-tion/ionisation,time-of-flight mass spectrometry,MALDI-TOF MS)technique to screen the differential methylation of CpG sites with lung cancer and analyze its correlation with clinicopathological characteristics and EGFR mutant subtype.Results The methylation levels of EGFR were significantly lower in the lung cancer group than in the normal control,EGFR#29 CpG_2,EGFR#30 CpG_8.9.10.11,CpG_25,CpG_40.41.42methylation of lung cancer group(25.7％±2.1％,4.6％±0.4％,2.8％±0.7％,4.40±0.4％)were low-er than the normal control(36.5％±4.6％,6.0％±0.6％,4.8％±0.9％,7.1％±0.4％),the difference was statistically signifi-cant(P＜0.05).No significant differences in the methylation levels within EGFR CpGs were associated with age,cancer differentiation,disease location,TNM stage with lung cancer(P＞0.05).The methylation level of EGFR#30 CpG_25 in the female group was signifi-cantly lower than that in the male group(P＜0.05),and the methylation level of EGFR#30 CpG_25 in the smoking group was higher than that in the non-smoking group(P＜0.05),the methylation level of EGFR#30 CpG_8.9.10.11 in the lymph node metastasis group was significantly lower than that in the non-lymph node metastasis(P＜0.01).There was no significant difference in methylation of EGFR between EGFR 19del and 21 L858R mutant subtypes.Conclusion Our findings suggest that EGFR methylation may be associated with the occurrence,development,gender,and smoking history of patients with EGFR driving gene-positive lung cancer,which may serve as a biomarker for the diagnosis and prognosis prediction of lung cancer.

Objective:To compare the biomechanical properties of the modified and traditional iliolumbar pedicle screw-rod system for the fixation of vertically unstable posterior pelvic ring dislocation.Methods:The CT images of the lumbar spine, pelvis, and femur from a 31-year-old healthy male volunteer were used to establish models of posterior pelvic ring dislocation and unilateral and bilateral fixation models of modified and traditional iliolumbar pedicle screw-rod systems with the three-dimensional virtual software using the finite element method. By restraining the distal ends of the femurs of both sides and applying a vertical load of 500 N to the L 1 vertebral body, the maximum stress of the intact pelvis model and the stiffness of the posterior pelvic ring dislocation model were evaluated to validate the reliability of the models. After the validation of the models, a follower load of 400 N was applied in the direction of the L 1 vertebral curve to simulate the upper body weight; the maximum vertical displacement, the maximum posterior displacement, the maximum right displacement, and the overall maximum displacement of the sacrum of the modified and traditional iliolumbar pedicle screw-rod system models were compared. Additionally, stress distributions of the implant and the pelvis were also compared. Results:(1) The maximum stress of the intact pelvis model was 22.0 MPa, with the stiffness of the posterior pelvic ring dislocation model as 180.03 N/mm. (2) In terms of the maximum vertical displacement of the sacrum, the bilateral modified iliolumbar pedicle screw-rod system was similar to the bilateral traditional iliolumbar pedicle screw-rod system. However, the maximum posterior displacement, the maximum right displacement, and the overall maximum displacement of the sacrum in the bilateral modified iliolumbar pedicle screw-rod system were 0.57 mm, 0.19 mm and 0.68 mm respectively, which were markedly smaller than those of the bilateral traditional iliolumbar pedicle screw-rod system (1.38 mm, 0.26 mm and 1.41 mm respectively). (3) The maximum vertical displacement, the maximum posterior displacement, the maximum right displacement, and the overall maximum displacement of the sacrum in the unilateral modified iliolumbar pedicle screw-rod system were 0.70 mm, 0.73 mm, 0.17 mm and 0.88 mm respectively, which were markedly smaller than those of the unilateral traditional iliolumbar pedicle screw-rod system (1.17 mm, 2.21 mm, 0.31 mm and 2.29 mm respectively). (4) The stress of the implant was concentrated on the connecting rod between the iliac pedicle screw-rod and the lumbar pedicle screw-rod in the modified iliolumbar pedicle screw-rod system, but it was mainly concentrated on the connecting rod at the upper edge of the iliac screw in the traditional iliolumbar pedicle screw-rod system. The maximum stress of the implant was 340.84 MPa in the bilateral modified iliolumbar pedicle screw-rod system, which was 30.4% less than that in the bilateral traditional iliolumbar pedicle screw-rod system (489.77 MPa), and was 351.23 MPa in the unilateral modified iliolumbar pedicle screw-rod system, which was 15.5% less than that in the unilateral traditional iliolumbar pedicle screw-rod system (415.82 MPa). (5) In both modified and traditional iliolumbar pedicle screw-rod systems, the stress of the pelvis was concentrated on the contact area between the iliac screw and the cortical bone. The maximum stress of the pelvis was 19.21 MPa in the bilateral modified iliolumbar pedicle screw-rod system, which was 78.1% less than that in the bilateral traditional iliolumbar pedicle screw-rod system (87.59 MPa), and was 39.91 MPa in the unilateral modified iliolumbar pedicle screw-rod system, which was 70.4% less than that in the unilateral traditional iliolumbar pedicle screw-rod system (134.98 MPa).Conclusion:Compared with the traditional iliolumbar pedicle screw-rod system, the modified iliolumbar pedicle screw-rod system can effectively reduce the displacements of all directions in vertically unstable posterior pelvic ring dislocation and significantly reduce the stress of the implant and the pelvis, thus having good biomechanical stability.

Following the principles of evidence-based medicine,in accordance with the structure and drafting rules of standardized documents,based on literature research,according to the characteristics of chronic cough in children and issues that need to form a consensus,the TCM Guidelines for Diagnosis and Treatment of Chronic Cough in Children was formulated based on the Delphi method,expert discussion meetings,and public solicitation of opinions.The guideline includes scope of application,terms and definitions,eti-ology and diagnosis,auxiliary examination,treatment,prevention and care.The aim is to clarify the optimal treatment plan of Chinese medicine in the diagnosis and treatment of this disease,and to provide guidance for improving the clinical diagnosis and treatment of chronic cough in children with Chinese medicine.

This article interprets the research progress and clinical applications of weed pollen allergen components as outlined in the European Academy of Allergy and Clinical Immunology (EAACI) guidelines on Molecular Allergology User′s Guide 2.0. The significance of this interpretation lies not only in emphasizing the analysis of patients′ sensitization patterns through advanced allergen component resolved diagnostics (CRD) but also in providing new research perspectives for exploring the unique features of weed pollen allergy in China. The complexity and diversity of weed pollen allergy, including its distribution and prevalence in different geographical regions, the characteristics of allergen component protein families, and their clinical significance, all require in-depth investigation. This interpretation aims to enhance the comprehensive understanding of allergen components in weed pollen allergy among relevant professionals, with the expectation of achieving outstanding progress in diagnosis and treatment. The ultimate goal is to develop more personalized and precise treatment strategies for patients with weed pollen allergy and those reacting to pan-allergens.

Objective:To investigate the effects of subcutaneous immunotherapy (SCIT) on patients′ immune markers and metabolic levels in the early stage of allergen treatment, and to gain insight into the role of SCIT in regulating immune responses and metabolic levels, so as to provide reference data for the further discovery of potential biomarkers.Methods:A longitudinal study was used to include 40 subjects who underwent SCIT with dust mite allergens in the Department of Pediatrics of the First Affiliated Hospital of Guangzhou Medical University between November 2017 and February 2022, including 20 subjects each of single mite subcutaneous immunotherapy (SM-SCIT) and double mite subcutaneous immunotherapy (DM-SCIT). In this study, levels of dust mite allergen-specific antibodies and polyunsaturated fatty acid metabolism were measured before and 12 months after treatment, while pulmonary function tests were performed. The therapeutic effects of the patients were followed up by visual analogue scale (VAS), asthma control test (ACT) and total medication scores (TMS). The results were statistically analyzed using t-test and Mann-Whitney U-test. Results:After 12 months of treatment with SCIT, both groups showed a significant decrease in total VAS score (SM-SCIT: Z=-2.298, P<0.05; DM-SCIT: Z=-3.411, P<0.001); total ACT score (SM-SCIT: Z=-2.054, P<0.05; DM-SCIT: Z=-2.014, P<0.05) and total medication scores (SM-SCIT: Z=-3.799, P<0.000 1; DM-SCIT: Z=-3.474, P<0.001) were significantly higher, in addition to significantly higher MMEF75/25 values in the DM-SCIT group ( t=-2.253, P<0.05). There was no significant change in sIgE in the SM-SCIT group ( P>0.05), and the sIgG4 levels of the Der p, Der f, p 1, p 2, f 2, and p 21 fractions were significantly elevated ( Z=-2.651, -3.771, -2.949, -2.912, -2.725, -2.128, and -3.285, respectively, all P<0.05); The sIgE of Der p 2, f 2, p 7 and p 23 fractions( Z=-2.651, -3.771, -2.949, -2.912, -2.725, -2.128, -3.285, all P<0.05) and the sIgG4 levels of the Der p, Der f, p 1, p 2, f 1, f 2, p 10, p 21 and p 23 fractions ( Z=-3.808, -3.845, -3.061, -2.688, -2.464, -3.211, -2.371, -2.091, -2.427, all P<0.05) of the DM-SCIT group were significantly elevated. Metabolomics analysis showed that arachidonic acid, docosahexaenoic acid, docosapentaenoic acid, eicosapentaenoic acid, 5, 9, 12-octadecatrienoic acid, 5(S)-hydroxylated eicosatetraenoic acid, and dihomo-gamma-linolenic acid were significantly elevated at the beginning of the treatment period after SM-SCIT treatment ( Z of -2.191, -2.497, -1.988, -2.090, -2.19, -2.803, -2.073, all P<0.05); 5(S)-hydroxylated eicosatetraenoic acid showed elevated and alpha-linolenic acid, eicosadienoic acid, and eicosapentaenoic acid were significantly decreased in the DM-SCIT group after treatment ( Z=-1.988, -2.090, -2.497, -1.988, respectively, all P<0.05). Correlation analysis showed that arachidonic acid was significantly negatively correlated with changes in dust mite-specific IgG4 ( r=-0.499, P<0.05), and that alpha-linolenic acid, 5, 9, 12-octadecatrienoic acid, and eicosapentaenoic acid were positively correlated with the ΔsIgG4 of the dust mite der p 2 ( r=0.451, 0.420, 0.474, respectively; all P<0.05). Conclusion:Significant changes in allergen-specific antibody levels and polyunsaturated fatty acid metabolism levels occur during SCIT, and the two may interact and influence each other.

Furry animal allergens, particularly cat and dog hair and dander, are common allergens in indoor environments, affecting the health of people world widely. Key sensitizing components such as Fel d 1 from cats and Can f 1 from dogs have been extensively studied and identified by the scientific community. Component resolved diagnosis (CRD) technology in modern diagnostic methods provides an accurate way to identify and distinguish these components, which is extremely important for the prevention of furry animal allergies and the formulation of personalized treatment strategies. To enhance the understanding of furry animal component diagnosis and promote the alignment of the Chinese discipline of allergology with international standards, this article interprets and explains the content of the "Molecular Allergology User′s Guide 2.0" recently released by the European Academy of Allergy and Clinical Immunology. It focuses on the epidemiological characteristics of furry animal components, the diversity of allergen protein families, and their clinical diagnosis and management.