BACKGROUND:Research has shown that vanillic acid has anti-inflammatory and anti-oxidative stress effects,but it is unclear whether it has a protective effect on endplate chondrocytes.OBJECTIVE:To explore the effect and mechanism of vanillic acid on endplate chondrocytes under inflammatory microenvironment.METHODS:(1)Primary endplate chondrocytes were isolated from the intervertebral disc of SD rats and identified by toluidine blue staining and collagenⅡ immunofluorescence.(2)The CCK-8 assay was employed to detect the effects of interleukin-1β and vanillic acid on the proliferation activity of endplate chondrocytes,in order to determine the concentration of vanillic acid for subsequent cell treatment.(3)An inflammatory microenvironment was simulated by adding 10 ng/mL interleukin-1β to the culture medium,and the endplate chondrocytes were treated with low,medium,and high mass concentrations of vanillic acid.The expression levels of inflammatory markers and extracellular matrix proteins were detected by western blot assay and immunofluorescence.(4)The expression of nuclear factor κB signaling pathway-related proteins was detected by western blot assay.RESULTS AND CONCLUSION:(1)The morphology of endplate chondrocytes in adherent culture was pike or triangular in shape,positive for toluidine blue staining and immunofluorescence for collagen Ⅱ,indicating that the experimentally extracted cells were endplate chondrocytes.(2)The CCK-8 assay results showed that treatment with 2.5,5,10,and 20 μg/mL vanillic acid for 24 hours did not significantly inhibit the proliferation of endplate chondrocytes.Compared with the interleukin-1β group,the viability of endplate chondrocytes treated with 5,10,and 20 μg/mL vanillic acid for 24 hours was significantly increased(P＜0.05).Therefore,5,10,and 20 μg/mL vanillic acid was selected as the low,medium,and high dose groups for subsequent treatment of endplate chondrocytes.(3)Compared with the model group(complete medium containing 10 ng/mL interleukin-1β),the expression of NOD-like receptor thermal domain associated protein 3(NLRP3),matrix metalloproteinase 13,matrix metalloproteinase 3,and tumor necrosis factor alpha protein in the endplate chondrocytes of the low,medium,and high doses of vanillic acid groups were significantly reduced(P＜0.05).(4)Compared with the model group,the protein expression of aggrecan and collagen Ⅱ in the endplate chondrocytes of the low,medium,and high dose groups of vanillic acid significantly increased(P＜0.05).(5)Compared with the model group,the protein expression of phospho-nuclear factor κB and phospho-inhibitor of nuclear factor κB in the endplate chondrocytes of the low,medium,and high dose groups of vanillic acid was significantly reduced(P＜0.05).(6)The above results indicate that vanillic acid may alleviate the inflammatory response and extracellular matrix degradation induced by interleukin-1β in rat endplate chondrocytes by inhibiting the activation of the nuclear factor κB signaling pathway.

Objective To develop an instrument for predicting postoperative constipation risks in patients with osteoporotic thoracolumbar fracture(OTLF)who have undergone percutaneous kyphoplasty(PKP).Methods A total of 858 OTLF patients who underwent PKP surgery between January 2020 and December 2024 were enrolled.The patients were randomly assigned to a training set(n=600)and a validation set(n=258)in a 7∶3 ratio.According to whether the patients had postoperative constipation,the training set was divided into a constipation group(n=205)and a non-constipation group(n=395),and the validation set was divided into a constipation group(n=90)and a non-constipation group(n=168).Logistic regression analysis was conducted to analyze the factors influencing postoperative constipation in OTLF patients after PKP,and a nomogram model was constructed accordingly.The receiver operating characteristic(ROC)curve and the calibration curve of the model were plotted,and the Hosmer-Lemeshow test for goodness of fit was performed.Results A total of 205 OTLF patients(34.17%)in the training set and 90 OTLF patients(34.88%)in the validation set experienced constipation after PKP.Univariate analysis revealed significant differences between the constipation and non-constipation groups in terms of operative time,postoperative water intake,time to first postoperative meal,postoperative bed rest time,the levels of Bifidobacterium,Lactobacillus,Enterococcus,and Enterobacter,the Nutrition Risk Screening 2002(NRS-2002)score,and the levels of sodium,potassium,and HbA1c(P<0.05).Least absolute shrinkage and selection operator(LASSO)regression was performed and operative time,time to first postoperative meal,the levels of Bifidobacterium,Lactobacillus,Enterococcus,and Enterobacter,the NRS-2002 score,and the levels of sodium,potassium,and HbA1c were identified as candidate predictors.Multivariate logistic analysis showed that the time to first postoperative meal,the levels of Bifidobacterium and Lactobacillus,the NRS-2002 score,and the levels of sodium and HbA1c were influencing factors of postoperative constipation in OTLF patients(P<0.05).The ROC curves showed that the area under the curve(AUC)of the training set was 0.842(95%CI:0.793-0.892),while that of the validation set was 0.860(95%CI:0.830-0.889).The calibration curves demonstrated good agreement between the prediction curve and the standard curve in both the training set and the validation set.Conclusion The time to the first postoperative meal,the NRS2002 score,and the levels of Bifidobacterium,Lactobacillus,sodium,and HbA1c are influencing factors of post-PKP constipation in OTLF patients.The nomogram model built based on these factors exhibited good predictive performance.

Objective:To collect registered clinical research plans on TCM characteristic therapies for treating cervical spondylosis; To explore their research registration status; To provide references for future clinical trial registration and implementation.Methods:Clinical research on TCM characteristic therapies for treating cervical spondylosis was retrieved from ChiCTR, ITMCTR and Clinical Trials. gov from the establishment of the databases to July 1, 2024. Excel 2019 was used to conduct descriptive statistics on registration time, registration area and institution, funding source, research type and design scheme, research participation center and sample size, cervical spondylosis type, intervention measures, outcome indicators, reporting quality, research openness and methodological application.Results:A total of 138 clinical trials for the TCM treatment of cervical spondylosis were included, of which 136 were registered by domestic researchers in 22 provincial-level administrative regions. The top three in terms of registration numbers were Shanghai, Guangdong Province, and Beijing. Additionally, 2 were registered by foreign researchers in Egypt and Malaysia. The main sources of funding were 50 local finances, followed by 26 hospital subsidies and 18 national finances. The intervention research accounted for the largest proportion of research types, with 123 items (89.13%). The research center mainly focused on single center studies (98 projects). Most randomized controlled trials (115 trials) described randomization methods, while a small number of randomized controlled trials (50 trials) indicated blinding. The intervention measures were mostly combined with TCM therapy, and the outcome indicators were mainly efficacy indicators, with fewer safety indicators.Conclusions:At present, clinical trial registrations for TCM treatment of cervical spondylosis are increasing, but issues remain, such as poor study design, uneven distribution, and incomplete information. It is recommended to refine registration details, optimize study protocols, and promote high-quality clinical research.

BACKGROUND:Research has shown that vanillic acid has anti-inflammatory and anti-oxidative stress effects,but it is unclear whether it has a protective effect on endplate chondrocytes.OBJECTIVE:To explore the effect and mechanism of vanillic acid on endplate chondrocytes under inflammatory microenvironment.METHODS:(1)Primary endplate chondrocytes were isolated from the intervertebral disc of SD rats and identified by toluidine blue staining and collagenⅡ immunofluorescence.(2)The CCK-8 assay was employed to detect the effects of interleukin-1β and vanillic acid on the proliferation activity of endplate chondrocytes,in order to determine the concentration of vanillic acid for subsequent cell treatment.(3)An inflammatory microenvironment was simulated by adding 10 ng/mL interleukin-1β to the culture medium,and the endplate chondrocytes were treated with low,medium,and high mass concentrations of vanillic acid.The expression levels of inflammatory markers and extracellular matrix proteins were detected by western blot assay and immunofluorescence.(4)The expression of nuclear factor κB signaling pathway-related proteins was detected by western blot assay.RESULTS AND CONCLUSION:(1)The morphology of endplate chondrocytes in adherent culture was pike or triangular in shape,positive for toluidine blue staining and immunofluorescence for collagen Ⅱ,indicating that the experimentally extracted cells were endplate chondrocytes.(2)The CCK-8 assay results showed that treatment with 2.5,5,10,and 20 μg/mL vanillic acid for 24 hours did not significantly inhibit the proliferation of endplate chondrocytes.Compared with the interleukin-1β group,the viability of endplate chondrocytes treated with 5,10,and 20 μg/mL vanillic acid for 24 hours was significantly increased(P＜0.05).Therefore,5,10,and 20 μg/mL vanillic acid was selected as the low,medium,and high dose groups for subsequent treatment of endplate chondrocytes.(3)Compared with the model group(complete medium containing 10 ng/mL interleukin-1β),the expression of NOD-like receptor thermal domain associated protein 3(NLRP3),matrix metalloproteinase 13,matrix metalloproteinase 3,and tumor necrosis factor alpha protein in the endplate chondrocytes of the low,medium,and high doses of vanillic acid groups were significantly reduced(P＜0.05).(4)Compared with the model group,the protein expression of aggrecan and collagen Ⅱ in the endplate chondrocytes of the low,medium,and high dose groups of vanillic acid significantly increased(P＜0.05).(5)Compared with the model group,the protein expression of phospho-nuclear factor κB and phospho-inhibitor of nuclear factor κB in the endplate chondrocytes of the low,medium,and high dose groups of vanillic acid was significantly reduced(P＜0.05).(6)The above results indicate that vanillic acid may alleviate the inflammatory response and extracellular matrix degradation induced by interleukin-1β in rat endplate chondrocytes by inhibiting the activation of the nuclear factor κB signaling pathway.

Objective To explore expression and significance in serum levels of soluble receptor for advanced glycation end products (sRAGE) and interleukin-36 (IL-36) in patients with Hashimoto's thyroiditis (HT). Methods A total of 180 individuals suspected of HT were enrolled as study subjects and divided into HT group (n=137) and non-HT group (n=43) based on final diagnostic outcomes. Within the HT group, patients were further divided into serum immunoglobulin G4 (IgG4)-related HT group (IgG4≥135 mg/dL) and non-IgG4-related HT group (IgG4 < 135 mg/dL) according to serum IgG4 levels. Serum levels of sRAGE and IL-36γ were measured in all subjects using enzyme-linked immunosorbent assay (ELISA). Receiver operating characteristic (ROC) curves were used to analyze the diagnostic value of serum sRAGE and IL-36γ levels forHT and its IgG4 phenotype. Results Serum sRAGE levels were significantly lower in the HT group compared to the non-HT group, while serum IL-36γ levels were significantly higher (P < 0.001). Patients in the IgG4-related HT group had significantly lower serum sRAGE levels and higher serum IL-36γ levels compared to those in the non-IgG4-related HT group (P < 0.001). ROC curve analysis revealed that the combined diagnosis of serum sRAGE and IL-36γ for HT had an area under the curve (AUC) of 0.900 (95%CI, 0.846 to 0.939), demonstrating significantly higher diagnostic efficiency than either indicator (P < 0.05). For the IgG4 phenotype in HT patients, the combined AUC of serum sRAGE and IL-36γ was 0.897 (95%CI, 0.934 to 0.943), which also significantly higher than individual indicator (P < 0.05). Conclusion Serum sRAGE levels are significantly decreased, while serum IL-36γ levels are significantly increased in HT patients. Combined detection of serum sRAGE and IL-36γ levels can effectively diagnose HT and assess the IgG4 phenotype.

Objective To explore the intestinal flora distribution characteristics in patients with Hashimoto's thyroiditis(HT)and analyze the correlation between intestinal flora changes and HT and the predictive value on the occurrence of HT,so as to provide reference for clinical diagnosis and treatment of HT.Methods A total of 120 patients with HT admitted to Qinhuangdao Traditional Chinese Medicine Hospital from June 2022 to June 2023 were included in observation group,and 30 healthy physical examination subjects during the same period were enrolled as control group.Both groups performed intestinal flora detection and thyroid function detection.The α diversity and the relative abundance of intestinal flora at phylum level were compared between the two groups and among HT patients with different thyroid functions,and the correlation between α diversity of intestinal flora and relative abundance of intestinal flora at phylum level,serum levels of anti-thyroglobulin antibody(TgAb)and thyroid peroxidase antibody(TPOAb)and thyroid function was analyzed by spearman correlation analysis.Receiver operating characteristic(ROC)curve was used to analyze the predictive efficiency of α diversity and relative abundance of intestinal flora at phylum level on HT.Results Compared with control group,the intestinal flora α diversity indexes of shannon-wiener diversity index(shannon)and simpson diversity index(simpson)were lower in observation group[4.66±0.61 vs 5.21±0.46,0.89±0.06 vs 0.93±0.07],and the differences were statistically significant(r=4.617,3.156,P＜0.05).The relative abundance of Bacteroidetes in observation group was lower than that in control group(52.31％±2.02％vs 58.59％±2.11％),while the relative abundances of Firmicutes,Proteobacteria,Fusobacteria,Actinobacteria and Verrucomicrobia were higher than those in control group(37.15％±2.43％vs 33.46％±2.56％,3.36％±0.26％vs 2.79％±0.19％,1.02％±0.09％vs 0.65％±0.11％,1.01％±0.12％vs 0.57％±0.13％,0.82％±0.11％vs 0.41％±0.09％),and the differences were statistically significant(t=15.096,7.360,11.267,19.231,17.665,18.882,all P＜0.001).The levels of serum TgAb(169.12±10.23 IU/ml vs 18.59±1.78 IU/ml)and TPOAb(56.43±12.11 IU/ml vs 2.51±0.57 IU/ml)in observation group were higher compared with those in control group,and the differences were statistically significant(t=80.097,24.319,all P＜0.001).Among 120 HT patients in this study,there were 89 cases with normal thyroid function and 31 cases with abnormal thyroid function.The intestinal flora α diversity indexes of shannon(4.49±0.64 vs 4.83±0.58),and simpson(0.87±0.07 vs 0.91±0.05)in patients with abnormal thyroid function were lower than those in normal patients,and the differences were statistically significant(t=2.736,3.439,all P＜0.05).The relative abundance of Bacteroides was lower(49.10％±1.99％vs 55.52％±2.05％),while the relative abundances of Firmicutes,Proteobacteria,Fusobacteria,Actinobacteria and Verrucomicrobia were higher than those in normal people(38.87％±2.60％vs 35.43％±2.26％,3.60％±0.26％vs 3.12％±0.19％,1.15％±0.09％vs 0.89％±0.11％,1.16％±0.13％vs 0.86％±0.11％,0.97％±0.13％vs 0.67％±0.09％)],and the differences were statistically significant(t=15.128,7.016,9.849,14.576,12.464,14.148,all P＜0.001).Spearman correlation analysis showed that the intestinal flora α diversity indexes(shannon and simpson)and relative abundance of Bacteroides in patients with HT were negatively correlated with serum TgAb and TPOAb levels(r=-0.436,-0.427,-0.402;-0.419,-0.456,-0.416,all P＜0.001),but were positively correlated with thyroid function(r=0.401,0.409,0.487,all P＜0.001).The relative abundances of Firmicutes,Proteobacteria,Fusobacteria,Actinobacteria and Verrucomicrobia were positively correlated with serum TgAb and TPOAb levels(r=0.411,0.441,0.447,0.421,0.447;0.425,0.415,0.438,0.402,0.469,all P＜0.001),but were negatively correlated with thyroid function(r=-0.412,-0.417,-0.475,-0.463,-0.471,all P＜0.001).ROC curve analysis revealed that the best cut-off value,the area under curve(AUC),sensitivity,specificity,95％confidence interval(95％CI)and Youden index in shannon,simpson,combined detection of simpson and shannon,and the combined detection of relative abundance of intestinal bacteria at phylum level for predicting HT were 4.677,0.749,49.17％,96.67％,95％CI(0.672～0.816)and 0.458;0.940,0.644,80.00％,53.33％,95％CI(0.562～0.721)and 0.333;0.752,85.83％,50.00％,95％CI(0.664～0.810)and 0.462;0.743,96.67％,46.67％,95％CI(0.665～0.811)and 0.548,respectively Conclusion HT had intestinal flora imbalance which was related to thyroid cell destruction and thyroid dysfunction in patients with HT.Intestinal flora detection has predictive efficiency on HT.

OBJECTIVE To optimize the formulation for sustained-release particles of matrine(MAT) and paeonol(PAE) and to primarily evaluate their quality. METHODS The sustained-release particles were fabricated by the extrusion-spheronization method. Single factor analysis was conducted on the main sustained-release materials and their dosage, auxiliary sustained-release materials, and the total amount and proportion of mixed sustained-release materials, using the cumulative release rates of MAT and PAE in vitro as evaluation indicators. By combining central composition design-response surface methodology, the formulation of sustained-release materials was optimized, and the effects of the total amount and mass ratio of ethyl cellulose(EC) and chitosan(CS) on the cumulative release rate and release synchronization of MAT and PAE were investigated. The formulation characteristics, in vitro release, and preliminary stability tests of MAT-PAE-SRPs were evaluated, and the release process kinetic equation was fitted. RESULTS The optimized formulation contained 23.5% EC and 17.1% CS as sustained-release materials. The yield, repose angle, bulk density and friability of final particles were 97.23%, 38.1°, 0.74 g·mL−1 and 0.74%, respectively. The particles showed sustained release pattern in various media and released faster in acidic media with its release percentage >90% at 12 h. The release profile of MAT was fitted best with first order equation, and that of paeonol with Higuchi equation. The formation of SRPs improved the stability of both drugs. CONCLUSION The sustained-release effect of MAT-PAE-SRPs prepared by sustained-release materials EC and CS is significant, and the cumulative release rate and release synchronization of MAT and PAE are good, which can provide reference for the research of dual loaded sustained-release formulations.

Objective:To compare the consistency of lymphoma multigene detection panels based on next-generation sequencing (NGS) with FISH detection of B-cell lymphoma gene rearrangement.Methods:From January 2019 to May 2023, fusion genes detected by lymphoma-related 413 genes that targeted capture sequencing of 489 B-cell lymphoma tissues embedded in paraffin were collected from Henan Cancer Hospital, and the results were compared with simultaneous FISH detection of four break/fusion genes: BCL2, BCL6, MYC, and CCND1. Consistency was defined as both methods yielding positive or negative results for the same sample. The relationship between fusion mutation abundance in NGS and the positivity rate of cells in FISH was also analyzed.Results:Kappa consistency analysis revealed high consistency between NGS and FISH in detecting the four B-cell lymphoma-related gene rearrangement ( P<0.001 for all) ; however, the detection rates of positive individuals differed for the four genes. Compared with FISH, NGS demonstrated a higher detection rate for BCL2 rearrangement, a lower detection rate for BCL6 and MYC rearrangement, and a similar detection rate for CCND1 rearrangement. No correlation was found between fusion mutation abundance in NGS and the positivity rate of cells in FISH. Conclusions:NGS and FISH detection of B-cell lymphoma gene rearrangement demonstrate overall good consistency. NGS is superior to FISH in detecting BCL2 rearrangement, inferior in detecting MYC rearrangement, and comparable in detecting CCND1 rearrangement.

Cancer toxin is a specific pathogenesis leading to the heterogeneity of breast cancer.The nature and virulence of the cancer toxin determine the differences in the heterogeneity of breast cancer,which can dynamically evolve over time and space,resulting in varying invasion abilities and characteristics of the tumor.Cancer cells in the primary lesion possess"toxicity"that targets specific organs for metastasis,and cancer toxins can influence the metastatic propensity of different types of breast cancer.Therefore,breast cancer treatment strategies based on the theory of cancer toxins emphasize the continuous eradication of the cancer toxin,focusing on differentiating its strength and nature,protecting unaffected areas first,identifying the state based on symptoms,and targeting accordingly to combat resistance arising from tumor heterogeneity.This article aims to provide a new theoretical basis for the treatment strategies of different types of breast cancer.

Objective:To explore the feasibility and effect of remote medical education model using online film reading training to improve the ability of ophthalmologists in the Xinjiang Uygur Autonomous Region (hereinafter referred to as "Xinjiang Region" ) in diagnosing fundus diseases.Methods:The three-level film reading training system of Xinjiang Production and Construction Corps system division hospital-Corps Hospital-Peking Union Medical College Hospital was established. From June 2022 to January 2023, 4 159 posterior color fundus images were continuously collected from Department of Ophthalmology of Xinjiang Corps Hospital and 4 divisional hospitals in the Corps medical system. Among them, hypertensive retinopathy, diabetic retinopathy, exudative age-related macular degeneration (AMD), atrophic AMD and retinal vein occlusion were 3 073, 651, 43, 186 and 206 cases, respectively. The images were divided into 3 rounds (first, second and last) according to the proportion of diseases. The doctors who participated in the training (hereinafter referred to as the "training") were 15 ophthalmologists from the Corps Hospital of Xinjiang Region and the division hospital of the Corps system. There were 7 male and 8 female. Age was (38.1±4.0) years. The titles of senior, deputy senior, intermediate and junior are 1, 6, 5 and 3 respectively; Bachelor's degree and master's degree are 13 and 2 respectively. The working time of fundus disease specialty was (9.6±3.3) years. The film reading system training was conducted before the first round of labeling, and after each round of film reading, the doctors of Peking Union Medical College Hospital gave feedback and explanation on the film reading results. The diagnostic consistency, sensitivity and specificity were compared by paired sample t test. Spearman or Pearson correlation analysis was conducted between the improvement of diagnostic level and professional title, education, age and working hours of ocular fundus disease. Results:All the participating doctors completed the first, second and last reading. After each round of film reading, the film reading summary was carried out for 2 hours. The average diagnostic agreement rates of participating physicians were 53.0%, 67.0% and 75.0%, respectively. The sensitivity and specificity were 0.38, 0.69, 054 and 0.66, 0.85, 0.96, respectively. There was significant difference between the first and last examination ( P<0.001). The sensitivity of the second reading was significantly higher than that of the first reading, and the sensitivity of the last reading was significantly lower than that of the second reading, with statistical significance ( P<0.05). The specificity of the second reading was significantly higher than that of the first reading, and the last reading was significantly higher than that of the second reading, with statistical significance ( P<0.05). There was no significant correlation ( P>0.05) between the improvement of diagnostic level of participating physicians and educational background ( Rho=0.07), professional title ( Rho=0.13), age ( r=0.20), and working time of ophthalmofundus disease specialty ( r=0.26). Conclusions:Relying on the three-level online telemedicine training, it can improve the ability of ophthalmologists in Xinjiang region to diagnose fundus diseases. The preliminary telemedicine education model has demonstrated potential for feasibility and effectiveness in remote areas with inadequate medical resources.