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.

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.

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 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.

With the rapid development of gene editing technology, the study of spermatogonial stem cells (SSCs) holds great significance in understanding spermatogenesis and its regulatory mechanism, developing transgenic animals, gene therapy, infertility treatment and protecting rare species. Biogenesis of lysosome-related organelles complex 1 subunit 1 (BLOC1S1) is believed to have anti-brucella potential. Exploring the impack of BLOC1S1 on goat SSCs not only helps investigate the ability of BLOC1S1 to promote SSCs proliferation, but also provides a cytological basis for disease-resistant breeding research. In this study, a BLOC1S1 overexpression vector was constructed by homologous recombination. The BLOC1S1 overexpression cell line of goat spermatogonial stem cells was successfully constructed by lentivirus packaging, transfection and puromycin screening. The overexpression efficiency of BLOC1S1 was found to be 18 times higher using real time quantitative PCR (RT-qPCR). Furthermore, the results from cell growth curve analysis, flow cytometry for cell cycle detection, and 5-ethynyl-2'-deoxyuridine (EdU) staining showed that BLOC1S1 significantly increased the proliferation activity of goat SSCs. The results of RT-qPCR, immunofluorescence staining and Western blotting analyses revealed up-regulation of proliferation-related genes (PCNA, CDK2, CCND1), and EIF2S3Y, a key gene regulating the proliferation of spermatogonial stem cells. These findings strongly suggest that the proliferative ability of goat SSCs can be enhanced through the EIF2S3Y/ERK pathway. In summary, this study successfully created a goat spermatogonial stem cell BLOC1S1 overexpression cell line, which exhibited improved proliferation ability. This research laid the groundwork for exploring the regulatory role of BLOC1S1 in goat spermatogonia and provided a cell platform for further study into the biological function of BLOC1S1. These findings also establish a foundation for breeding BLOC1S1 overexpressing goats.
Animals ; Male ; Goats ; Stem Cells ; Spermatogonia/metabolism* ; Cell Proliferation ; Flow Cytometry ; Testis/metabolism*

Objective: To investigate the status of insomnia, Internet addiction, and depressive symptoms among medical students and to analyze the effect of Internet addiction on insomnia and the mediating role of depressive symptoms, in order to provide a basis for the development of targeted interventions and measurements for medical students. Methods: A stratified whole group sampling method was used to select full-time college students from three medical universities in Anhui Province. The Chinese version of Insomnia Severity Index (ISI), Internet Addiction Test (IAT) scale and 9-item Patient Health Questionnaire (PHQ-9) were used to evaluate the symptoms of insomnia, Internet addiction and depressive in students. A multivariate Logistic regression analysis was used to explore the factors influencing insomnia among medical students and to analyze the relationship between insomnia with Internet addiction and depressive symptoms, respectively. Results: The overall rate of Internet addiction was 49.5%, depressive symptoms was 39.5%, insomnia was 18.6%. High academic stress, and the presence of surrounding people diagnosed with COVID-19 were associated with a higher risk of insomnia ( P <0.05). The higher the level of Internet addiction (mild, OR =2.60; moderate/severe, OR =4.21) and depression. (mild, OR =6.35; moderate/severe, OR =19.32), the higher the risk of insomnia. Mediated effect analysis showed that Internet addiction had a direct predictive effect ( β =0.02, P <0.05) on insomnia and also indirectly affected insomnia through depression (indirect effect=0.07，95% CI =0.06-0.08). Conclusion The detected rates of insomnia, Internet addiction and depressive symptoms are high among medical students in Anhui Province, and Internet addiction and depressive symptoms are risk factors for insomnia, which should be given more attention and appropriate interventions when necessary to improve their physical and mental health.

Objective:To evaluate the risk factors for adjacent vertebral compression fracture (AVCF) after fusion surgery of lumbar canal stenosis in the elderly.Methods:A retrospective cohort study was designed for 297 patients with lumbar canal stenosis who underwent fusion surgery in Hubei Provincial Hospital of Integrated Chinese and Western Medicine from January 2017 to December 2020. There were 42 males and 255 females, aged 68-85 years [(76.3±7.2)years]. The patients were divided into AVCF group ( n=67) and non-AVCF group ( n=230) according to with or without the occurrence of AVCF. The indicators recorded were the gender, age, body mass index, education level, number of combined basic diseases, course of the disease, preoperative bone mineral density, intraoperative bleeding volume, number of level fused during operation, postoperative blood transfusion volume, postoperative observation in the ICU, types of postoperative external fixation and time of external fixation. Univariate analysis was conducted to analyze the correlation between above risk factors and AVCF after fusion surgery of lumbar canal stenosis in the elderly. Multivariate Logistic regression analysis was used to determine the independent risk factors for the occurrence of AVCF in these patients. Results:In the univariate analysis, age, education level, preoperative bone mineral density, intraoperative bleeding volume, postoperative blood transfusion volume and postoperative observation in the ICU were correlated with AVCF after fusion surgery of lumbar canal stenosis in the elderly ( P<0.05 or 0.01), while there was no correlation of AVCF with gender, body mass index, number of combined basic diseases, course of the disease, number of level fused during operation, types of postoperative external fixation and time of external fixation (all P>0.05). Multivariate Logistic regression analysis showed that gender ( OR=4.02, 95% CI 1.35, 12.00, P<0.05), preoperative bone mineral density≤-2.5 SD ( OR=2.01, 95% CI 1.47, 2.75, P<0.01), intraoperative bleeding volume≥475 ml ( OR=1.01, 95% CI 1.00, 1.01, P<0.01) and postoperative blood transfusion volume≥434 ml ( OR=0.99, 95% CI 0.98, 1.00, P<0.01) were significantly associated with AVCF after fusion surgery of lumbar canal stenosis in the elderly. Conclusion:The female, bone mineral density≤-2.5 SD, intraoperative bleeding volume≥476 ml and postoperative blood transfusion volume≥434 ml are independent risk factors for AVCF after fusion surgery of lumbar canal stenosis in the elderly.

Based on the structure of combretastatin A-4 (CA-4), a microtubulin inhibitor, eight novel compounds were designed and synthesized by introducing different substituents into the benzimidazole backbone which substituted B ring of CA-4, and the structures were characterized by NMR and HRMS. Proliferation inhibition of six tumor cells including A549, HepG2, HCT-116, MCF-7, PC-3 and Siha was measured by MTT method.The effect of active compound on cell migration was evaluated by scratch test.Molecular docking technique was applied to investigate the interaction between the most active compound with the tubulin and PI3K kinases respectively.Compound 4e showed prominent inhibition against six strains of tumor cells, especially with the strongest inhibitory effect on Siha cells (IC50 = 12.18 ± 1.17 μmol/L).Moreover, compound 4e could effectively inhibit cell migration, which deserves further study.Molecular docking study showed that the binding energy to the tubulin of compound 4e was stronger than that of CA-4, and the affinity with PI3Ks displayed that the PI3Kδ subtype kinase was the strongest; its binding energy was -37.2 kJ/mol.This study lays a foundation for the development of anti-tumor drug based on PI3K and microtubulin.

Objective:To investigate the feasibility of reducing the radiation dose on coronary artery calcium score (CS) of virtual non-contrast (VNC) scanning in dual-layer spectral coronary CT angiography(CCTA).Methods:One hundred and twenty-two patients were examined on a dual-layer spectral detector CT scanner from March 2019 to August 2020. Volume CT dose index (CTDI vol), dose length product (DLP), effective dose ( E) were all evaluated for each patient. CS was calculated from both true non-contrast (TNC) and VNC images for left anterior descending (LAD), left circumflex (LCx), right coronary artery (RCA), and the total coronary artery (Total) by two radiologists independently. Pearson′s correlation coefficient was calculated for measuring the association between variables. The correction coefficients of each branch (λ LAD, λ LCx, and λ RCA) and the average correction coefficient (λ AVG) of the total coronary artery were obtained. The calibrated calcium score (CCS_VNC) was equal to λ multiplied by CS_VNC. The CS_TNC and CCS_VNC were compared using repeated oneway analysis of variance test. Correlation analyses for CS_TNC and CCS_VNC and agreement evaluation with Bland-Altman-Plots were performed. Results:The average effective doses in TNC, CCTA and total group were 0.69, 6.47 and 7.16 mSv, respectively. The effective dose was reduced by 10.6% and the scan time was reduced by 39% while using VNC images. There were significant differences among the CS_TNC and CS_VNC of LAD, LCx, RCA and Total ( t=6.75, 5.33, 4.99, 6.60, P< 0.05). Excellent correlations were observed between CS_VNC and CS_TNC ( R2 values were 0.929, 0.896, 0.958, and 0.918; λ values were 2.18, 1.18, 2.15, and 2.07, respectively). There were no significant statistically difference among the CS_TNC, CCS_VNC AVG, and CCS_VNC LAD/RCA of the LAD and RCA (all P> 0.05). The difference was statistically significant among the CS_TNC, CCS_VNC AVG, and CCS_VNC LCx of the LCx ( F=10.94, P<0.05). The paired comparison were performed in groups and the differences were statistically significant between the CS_TNC versus CCS_VNC AVGand CCS_VNC AVG versus CCS_VNC LCx ( t=3.31, 3.43, all P<0.05). There was no significant statistically difference between the CCS_VNC LCx and CCS_VNC AVG( P>0.05). Conclusions:It was feasible to accurately evaluate the CS_VNC from spectral data in comparison to TNC imaging, and to reduce the patient radiation dose and acquisition time.