OBJECTIVES: The core pathology of osteoporosis lies in bone resorption exceeding bone formation; thus, promoting osteogenesis is a key therapeutic strategy. The osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) forms the biological basis of bone formation. Astragaloside IV (A-IV), a major active component of Astragalus membranaceus, is known to enhance osteogenesis, but its precise molecular mechanisms remain unclear. This study aims to investigate the effects of A-IV on the proliferation and osteogenic differentiation of BMSCs from osteoporotic rats and to elucidate its molecular mechanism through the regulation of microRNA-21 (miR-21) and Notch2 expression. METHODS: After 1 week of adaptive feeding, mature female SD rats were randomly divided into a sham-operated (Sham) group (n=4) and an ovariectomized (OVX) group (n=8) to establish an osteoporosis model. Twelve weeks after surgery, BMSCs were isolated from femoral bone marrow and cultured. Cells were divided into a S-BMSCs group (from Sham), an O-BMSCs group (from OVX), and an A-BMSCs group (from OVX-derived BMSCs treated with A-IV). S-BMSCs and O-BMSCs were induced for osteogenic differentiation using osteogenic induction medium, whereas A-BMSCs were treated with A-IV before induction. Flow cytometry was used to identify mesenchymal stem cell surface markers (CD29) and hematopoietic stem cell marker (CD34) to confirm BMSC characteristics. Cell proliferation was assessed using the methyl thiazolyl tetrazolium (MTT) assay. Alizarin red staining was performed to quantify calcium nodule formation, and alkaline phosphatase (ALP) activity assays were used to evaluate osteogenic differentiation. Real-time reverse transcription PCR (real-time RT-PCR) was used to detect changes in osteogenic-related genes, runt-related transcription factor 2 (Runx2) and osteopontin (OPN), as well as miR-21 expression. Western blotting was performed to assess Runx2, OPN, and Notch2 protein expression. RESULTS: Flow cytometry confirmed that O-BMSCs retained the phenotypic characteristics of mesenchymal stem cells. A-IV significantly enhanced the proliferation of BMSCs from osteoporotic rats (P<0.05), increased ALP activity, and upregulated the mRNA and protein expression of Runx2 and OPN (P<0.05). Bioinformatic and experimental analyses demonstrated that miR-21 directly targeted Notch2. A-IV treatment increased miR-21 expression while suppressing Notch2 protein expression and inhibiting activation of the Notch signaling pathway (P<0.05). CONCLUSIONS Astragaloside IV promotes the osteogenic differentiation of BMSCs derived from osteoporotic rats by upregulating miR-21 expression and inhibiting the key Notch signaling protein Notch2, thereby relieving the Notch2-mediated suppression of osteogenesis.
Animals ; Triterpenes/pharmacology* ; Saponins/pharmacology* ; Osteogenesis/drug effects* ; MicroRNAs/metabolism* ; Rats, Sprague-Dawley ; Female ; Cell Differentiation/drug effects* ; Mesenchymal Stem Cells/drug effects* ; Signal Transduction/drug effects* ; Osteoporosis/pathology* ; Rats ; Cells, Cultured ; Receptor, Notch2/metabolism* ; Receptors, Notch/metabolism* ; Ovariectomy ; Cell Proliferation/drug effects*

OBJECTIVES: To investigate the role of METTL3 and FOXO3 in anthracycline resistance in acute myeloid leukemia (AML) cells. METHODS: Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and transcriptome sequencing (RNA-seq) were performed in anthracycline-resistant and sensitive HL60 and K562 cells with lentivirus-mediated knockdown or overexpression of METTL3 and FOXO3. TCGA and GSE6891 datasets were used for analysis of the clinical and gene expression data of AMI patients. FOXO3 expressions at the mRNA and protein levels in the transfected cells were detected with RT-qPCR and Western blotting, and the changes in cell proliferation and apoptosis were evaluated using CCK8 assay and flow cytometry; the expression of m⁶A-modified mRNA and mRNA stability of FOXO3 was detected analyzed using MeRIP-qPCR and RT-qPCR. Functional enrichment analysis of the differential genes in the transfected cells was performed. RESULTS: Differential gene analysis in anthracycline-resistant versus sensitive AML cells and in cells with METTL3 knockdown revealed the enrichment in FoxO and autophagy pathways (P<0.05), and the anthracycline-resistant cells showed significantly increased m⁶A modification of FOXO3. FOXO3 expression was positively correlated with METTL3 expression. METTL3 knockdown significantly reduced FOXO3 mRNA stability and its protein levels in anthracycline-resistant AML cells, which exhibited higher m6A-modified FOXO3 expression levels than their sensitive counterparts. Database analysis, Kaplan-Meier analysis and RT-qPCR results suggested that a high FOXO3 expression was associated with a poor prognosis of AML patients. In anthracycline-resistant AML cells expressing higher FOXO3 levels than the sensitive cells, lentivirus-mediated overexpression of FOXO3 significantly enhanced cell proliferation and suppressed cell apoptosis. Inhibiting autophagy using an autophagy inhibitor (Baf.A1) obviously enhanced the inhibitory effect of adriamycin on resistant AMI cells and cells overexpressing FOXO3. CONCLUSIONS METTL3 promotes FOXO3 expression via m6A modification, and FOXO3-driven autophagy contributes to anthracycline resistance in AML cells by enhancing cell proliferation and suppressing cell apoptosis.
Humans ; Forkhead Box Protein O3/genetics* ; Leukemia, Myeloid, Acute/genetics* ; Drug Resistance, Neoplasm ; Methyltransferases/genetics* ; Autophagy ; Anthracyclines/pharmacology* ; HL-60 Cells ; Apoptosis ; Cell Proliferation ; K562 Cells

Objective To evaluate the clinical characteristics of human immunodeficiency virus (HIV) infected patients with peripheral lung masses (PLMs), and to assess the diagnostic utility of contrast-enhanced ultrasound (CEUS) in differentiating benign and malignant PLMs. Methods A retrospective analysis was performed on the clinical data of 69 patients with PLM treated in Shanghai Public Health Clinical Center from January 2020 to December 2023. All patients underwent percutaneous biopsy, and were categorized into benign group (n＝36) and malignant group (n＝33). 25 patients were HIV-positive and 44 patients were HIV-negative. The clinical features and CEUS parameters in patients were compared across these groups. Results Patients with malignant masses were significantly older than those with benign masses (P＜0.05). In the malignant group, HIV-negative patients exhibited significantly larger tumor diameters compared to HIV-positive patients (P＜0.05); in the HIV-positive patients, no significant difference in tumor size was observed between benign and malignant masses. 19 patients underwent CEUS. 10 malignant masses, irrespective of HIV status (10 positive and 9 negative), commonly presented with indistinct margins, delayed enhancement, heterogeneous perfusion, and delayed peak enhancement on CEUS. 9 benign masses showed earlier peak enhancement compared to 10 malignant masses (P＜0.05); no significant differences were observed in the initiation and washout time of enhancement between benign and malignant masses. In HIV-positive patients, 5 benign masses frequently demonstrated discrepancies between CEUS findings and pathological results. Conclusions The clinical and CEUS characteristics were different between benign and malignant PLMs. However, CEUS shows limited accuracy in distinguishing benign and malignant PLMs, underscoring the need for pathological confirmation.

Objective To explore the problems arising from the safety content in the instructions of Chinese patent medicines containing bitter almonds and to make suggestions.Methods By searching the Chinese patent medicines containing bitter almonds in the"National Basic Medical Insurance,Work Injury Insurance and Maternity Insurance Drug List(2022)"of the Medication Assistant and Yaozhi Online,the article collated their primary functions,composition,adverse reactions,contraindi-cations to medication,warnings,and precautions.The article entered the collected information into the Excel 2021 version,analyzed the summarized data,and made suggestions for the safety warning information in the instructions of Chinese patent medicines con-taining bitter almonds based on the results of the statistics.Results Among the 1 381 kinds of patent medicines listed in the"National Basic Medical Insurance,Work Injury Insurance,and Maternity Insurance Drug List(2022)",there are 67 varieties of Chinese patent medicines containing bitter almonds,and there were missing safety contents in the collected instructions.Among them,only five types were clearly labeled with specific contents of warnings,17 types were clearly labeled with adverse reactions,24 types were clearly labeled with contraindications,and 58 types were clearly labeled with precautions.However,in terms of the content of bitter almonds and prescription ratio,the maximum daily dosage of 2 out of 67 Chinese patent medicines containing bit-ter almonds exceeded the dosage of the Chinese Pharmacopoeia,and the prescription ratio of 14 Chinese patent medicines excee-ded 10％;in terms of the combination and dosing,25 out of 67 Chinese patent medicines containing bitter almond were dosed with other poisonous traditional Chinese medicines,and the above medicines have high safety risks.The information on their safety should be increased,and their safety information should be increased.Conclusions The safety-related warnings in the instruc-tions for Chinese patent medicines containing bitter almonds are not comprehensive enough.It is recommended to strengthen the management of the whole life cycle of drugs,increase the warnings of level 1 high-risk varieties,and improve adverse reactions,contraindications,and precautions according to the Technical Guiding Principles for Revision of Safety Information in Marketed Chinese Patent Medicine Instructions(Trial).

Objective:Measure the global longitudinal strain (GLS) of the left ventricle in trauma patients by beside speckle tracking echocardiography to explore the role of STE -GLS in evaluating left ventricular systolic function in trauma patients, and then explore the clinical value of GLS in judging the prognosis of trauma patients.Methods:Trauma patients admitted to intensive care unit from September 1, 2020 to April 1, 2021 with an Injury Severity Score (ISS) of ≥ 16 points. were consecutively enrolled. Moreover, those patients who met the following criteria were selected as the research subjects: aged between 18 and 80 years old, had no serious underlying diseases in the past, the time from trauma onset to admission was within 24 hours, and were able to complete an echocardiogram examination within 24 hours after the onset of the disease. Exclude patients who are unable to complete the ultrasound examination within 24 hours after the onset of the disease, or those with poor image quality, or those complicated with severe heart diseases and systemic comorbidities. The left ventricular global longitudinal strain (GLS) was measured by bedside speckle tracking echocardiography. According to the GLS values they were divided into abnormal group (GLS> -15%) and normal group (GLS≤ -15%). Independent sample t-tests and chi-square tests were applied to conduct a comparative analysis of the clinical characteristics between the two groups of patients. Furthermore, multiple linear regression analysis was conducted to explore the correlation between STE-GLS and the duration of intensive care unit stay.Results:A total of 32 trauma patients were eligible for this study. One patient was found to have abnormal left ventricular systolic function (LVEF<50%) detected by conventional echocardiography, however speckle tracking echocardiography detected decreased left ventricular systolic function (GLS> -15%) in 13 Patients. Multiple linear regression analysis showed that the global longitudinal strain of left ventricle and serum high sensitivity troponin I were independent risk factors affecting the time of intensive care in trauma patients.Conclusions:Speckle tracking echocardiography (STE) is more sensitive than traditional echocardiography and can detect left ventricular systolic dysfunction early. STE-GLS is an independent risk factor affecting hospitalization time of trauma patients in intensive care unit. Clinically, STE-GLS and serum Hs-TnI can be combined to determine the prognosis of trauma patients.

Objective To explore the problems arising from the safety content in the instructions of Chinese patent medicines containing bitter almonds and to make suggestions.Methods By searching the Chinese patent medicines containing bitter almonds in the"National Basic Medical Insurance,Work Injury Insurance and Maternity Insurance Drug List(2022)"of the Medication Assistant and Yaozhi Online,the article collated their primary functions,composition,adverse reactions,contraindi-cations to medication,warnings,and precautions.The article entered the collected information into the Excel 2021 version,analyzed the summarized data,and made suggestions for the safety warning information in the instructions of Chinese patent medicines con-taining bitter almonds based on the results of the statistics.Results Among the 1 381 kinds of patent medicines listed in the"National Basic Medical Insurance,Work Injury Insurance,and Maternity Insurance Drug List(2022)",there are 67 varieties of Chinese patent medicines containing bitter almonds,and there were missing safety contents in the collected instructions.Among them,only five types were clearly labeled with specific contents of warnings,17 types were clearly labeled with adverse reactions,24 types were clearly labeled with contraindications,and 58 types were clearly labeled with precautions.However,in terms of the content of bitter almonds and prescription ratio,the maximum daily dosage of 2 out of 67 Chinese patent medicines containing bit-ter almonds exceeded the dosage of the Chinese Pharmacopoeia,and the prescription ratio of 14 Chinese patent medicines excee-ded 10％;in terms of the combination and dosing,25 out of 67 Chinese patent medicines containing bitter almond were dosed with other poisonous traditional Chinese medicines,and the above medicines have high safety risks.The information on their safety should be increased,and their safety information should be increased.Conclusions The safety-related warnings in the instruc-tions for Chinese patent medicines containing bitter almonds are not comprehensive enough.It is recommended to strengthen the management of the whole life cycle of drugs,increase the warnings of level 1 high-risk varieties,and improve adverse reactions,contraindications,and precautions according to the Technical Guiding Principles for Revision of Safety Information in Marketed Chinese Patent Medicine Instructions(Trial).

This study was performed to investigate the therapeutic effect of oroxylin A(OA)on chicken type Ⅱcollagen-induced arthritis(CIA)in mice and observe the changes of immune cells.The CIA model was established,and 40 mg/kg OA was intraperitoneally injected for 10 consecutive days from the 28th day.The mice were sacrificed at three different times during the administration period,and the joints were scored at each time point.HE staining was used to observe the pathology of the mouse ankle joint;flow cytometry was used to detect the changes of Th17,Treg and macrophages in spleen and inguinal lymph nodes;ELISA was employed to detect the expression levels of IL-1β,IL-18 and IL-6 in serum;and qRT-PCR was used to detect the expression levels of IL-1β,IL-18,IL-6 and TNF-β in spleen of mice.Data showed that on the 34th day after OA administration,the joint swelling of CIA mice was significantly relieved,the pathological score was decreased,and the inflammatory cell infiltration was decreased.Flow cytometry results showed that the proportion of Th17 cells and macrophages in the spleen and inguinal lymph nodes of CIA mice in OA group decreased,while the proportion of Treg cells increased.The results of ELISA and qRT-PCR showed that OA could inhibit the level of inflammation in CIA mice.In conclusion,OA can regulate the proportion of immune cells,inhibit the level of inflammation in CIA mice,and then relieve the symptoms of CIA mice.

Objective To establish in vitro diagnostic(IVD)reagents closed-loop management scheme for the whole process and implement,and optimize the IVD management.Methods Based on the status quo of IVD management in Beijing Hospital,the closed-loop management scheme of the whole IVD process was designed,such as IVD application,IVD selection,IVD subscription,IVD purchase,IVD receipt,IVD inbound and outbound,and IVD use.Key indicators were selected to establish an evaluation framework.Management data,procurement data and testing data before(2020)and after(2023)implementation of the program were collected and counted by means of interviews and consulting relevant information systems,and then process management,quality management and cost management were evaluated.Results The management process was optimized,the IVD purchase time was shortened by about 60％on average,the IVD purchase did not need manual processing,the order was split in real time and sent to the corresponding supplier,and the IVD collection time was shortened by about 75％on average.IVD and test quality management were satisfactory,supplier qualification perfection reached 100％,full marks were obtained in the inter-room quality assessment,and no IVD-related adverse events were observed.The cost management was effective,and the IVD procurement cost was reduced by about 15％.IVD cost fine management models were established for four items,namely,human immunodeficiency virus antibody assay,hepatitis C antibody assay,treponemal antibody assay and hepatitis B surface antigen quantitative assay.The average value of IVD loss was 0.07％～1.21％,and the standard deviation was 0.07％～0.66％.Conclusion The IVD whole process closed-loop management scheme is feasible,which can improve the efficiency of IVD management,ensure the quality of IVD and detection,reduce the cost of IVD,and refine the cost management work.

Objective To explore the value of deep learning models in predicting axillary lymph node(ALN)metastasis of breast cancer based on intratumoral and peritumoral dynamic contrast-enhanced magnetic resonance imaging(DCE-MRI).Methods A retrospective analysis was conducted on cases from Tianjin Medical University Cancer Hospital and Laoshan Branch of Affiliated Hospital of Qingdao University,involving a total of 850 lesions in 850 patients.The region of interest within the tumor was delineated at the largest area of the lesion on the first enhancement images and automatically expanded by 3 mm and 6 mm in a conformal fashion.Deep learning prediction models based on ResNet50 were developed via intratumoral,peritumoral,and intratumoral combined peritumoral models,respectively,and a comprehensive prediction model was developed by integrating semantic features of imaging reports.Cases from Tianjin Medical University Cancer Hospital were randomly divided into training and test cohorts in a 7∶3 ratio,while cases from Laoshan Branch of Affiliated Hospital of Qingdao University served as the external validation cohort.The area under the curve(AUC),accuracy,sensitivity,specificity,F1-score,and Brier-score were calculated,respectively.Results The model incorporating intratumoral,peritumoral(3 mm),and semantic features demonstrated the highest performance,with AUC of 0.801[95％confidence interval(CI)0.765-0.845],0.781(95％CI 0.745-0.817),and 0.752(95％CI 0.700-0.793)in the training cohort,test cohort,and external validation cohort,respectively,and there was no significant difference in AUC between combined model and intratumoral/peritumoral model,respectively,but it demonstrated the higher sensitivity and F1-score,and the lower Brier-score.Conclusion Incorporating peritumoral images into the conventional model based on intratumoral images enhanced the predictive ability of ALN metastasis in breast cancer.

Diarrhea is the most common clinical symptom of the digestive system,its pathogenesis may be related to intestinal infection,intestinal microecological instability,intestinal barrier dysfunction and so on.Sijunzi decoction has the effect of invigorating spleen and nourishing stomach,and supplementing qi and blood.In modern clinical practice,modified Sijunzi decoction combining with other prescriptions,Chinese medicine massage,acupuncture and western medicine attenuates synergies,improves the patients'quality of life,and is widely used in the treatment of diarrhea,but its mechanism has not yet been systematically clarified.Therefore,this article reviews the research progress of the mechanism and clinical application of Sijunzi decoction and its modified formula in the treatment of diarrhea,and provides reference for the mechanism research and prevention of Sijunzi decoction and its modified formula in the treatment of diarrhea.