OBJECTIVE: To prepare clinical-grade human umbilical cord-derived mesenchymal stem cells (hUC-MSC) with high expression of hematopoietic supporting factors and evaluate their stem cell characteristics. METHODS: Fetal umbilical cord tissues were collected from healthy postpartum women during full-term cesarean section. Wharton's jelly was mechanically separated and hUC-MSCs were obtained by explant culture method and enzyme digestion method in an animal serum-free culture system with addition of human platelet lysate. The phenotypic characteristics of hUC-MSCs obtained by two methods were detected by flow cytometry. The differences in proliferation ability between the two groups of hUC-MSCs were identified through CCK-8 assay and colony forming unit-fibroblast (CFU-F) assay. The differences in multilineage differentiation potential between the two groups of hUC-MSCs were identified through induction of adipogenic, osteogenic, and chondrogenic differentiation. The mRNA expression levels of hematopoietic supporting factors such as SCF, IL-3, CXCL12, VCAM1 and ANGPT1 in the two groups of hUC-MSCs were identified by real-time fluorescence quantiative PCR(RT-qPCR). RESULTS: The results of flow cytometry showed that hUC-MSCs obtained by the two methods both expressed high levels of CD73, CD90 and CD105, while lowly expressed CD31, CD45 and HLA-DR. The results of CCK-8 and CFU-F assay showed that the proliferation ability of hUC-MSCs obtained by explant culture method was better than those obtained by enzyme digestion method. The results of the triple lineage differentiation experiment showed that there was no significant difference in multilineage differentiation potential between the two grous of hUC-MSCs. The results of RT-qPCR showed that the mRNA expression levels of hematopoietic supporting factors SCF, IL-3, CXCL12, VCAM1 and ANGPT1 in hUC-MSCs obtained by explant cultrue method were higher than those obtained by enzyme digestion method. CONCLUSION Clinical-grade hUC-MSCs with high expression levels of hematopoietic supporting factors were successfully cultured in an animal serum-free culture system.
Humans ; Mesenchymal Stem Cells/metabolism* ; Umbilical Cord/cytology* ; Cell Differentiation ; Female ; Cell Proliferation ; Cells, Cultured ; Chemokine CXCL12/metabolism* ; Angiopoietin-1/metabolism* ; Vascular Cell Adhesion Molecule-1/metabolism* ; Stem Cell Factor/metabolism* ; Flow Cytometry ; Pregnancy

This study aimed to explore the effects and mechanisms of total extracts from Anthriscus sylvestris on pulmonary hypertension in rats. Sixty male SD rats were divided into normal(NC) group, model(M) group, positive drug sildenafil(Y) group, low-dose A. sylvestris(ES-L) group, medium-dose A. sylvestris(ES-M) group, and high-dose A. sylvestris(ES-H) group. On day 1, rats were intraperitoneally injected with monocrotaline(60 mg·kg~(-1)) to induce pulmonary hypertension, and the rat model was established on day 28. From days 15 to 28, intragastric administration of the respective treatments was performed. After modeling and treatment, small animal echocardiography was used to detect the right heart function of the rats. Arterial blood gas was measured using a blood gas analyzer. Hematoxylin and eosin(HE) staining and Masson staining were performed to observe cardiopulmonary pathological damage. Flow cytometry was used to detect apoptosis in the lung and myocardial tissues and reactive oxygen species(ROS) levels. Western blot was applied to detect the expression levels of transforming growth factor-β1(TGF-β1), phosphorylated mothers against decapentaplegic homolog 3(p-Smad3), Smad3, tissue plasminogen activator(t-PA), and plasminogen activator inhibitor-1(PAI-1) in lung tissue. A blood routine analyzer was used to measure inflammatory immune cell levels in the blood. Enzyme-linked immunosorbent assay(ELISA) was used to detect the expression levels of P-selectin and thromboxane A2(TXA2) in plasma. The results showed that, compared with the NC group, right heart hypertrophy index, right ventricular free wall thickness, right heart internal diameter, partial carbon dioxide pressure(PaCO_2), apoptosis in cardiopulmonary tissue, and ROS levels were significantly increased in the M group. In contrast, the ratio of pulmonary blood flow acceleration time(PAT)/ejection time(PET), right cardiac output, change rate of right ventricular systolic area, systolic displacement of the tricuspid ring, oxygen partial pressure(PaO_2), and blood oxygen saturation(SaO_2) were significantly decreased in the M group. After administration of the total extract of A. sylvestris, right heart function and blood gas levels were significantly improved, while apoptosis in cardiopulmonary tissue and ROS levels significantly decreased. Further testing revealed that the total extract of A. sylvestris significantly decreased the levels of interleukin-1β(IL-1β), interleukin-6(IL-6), and PAI-1 proteins in lung tissue, while increasing the expression of t-PA. Additionally, the extract reduced the levels of inflammatory cells such as leukocytes, lymphocytes, granulocytes, and monocytes in the blood, as well as the levels of P-selectin and TXA2 in plasma. Metabolomics results showed that the total extract of A. sylvestris significantly affected metabolic pathways, including arginine biosynthesis, tyrosine metabolism, and taurine and hypotaurine metabolism. In conclusion, the total extract of A. sylvestris may exert an anti-pulmonary hypertension effect by inhibiting the TGF-β1/Smad3 signaling pathway, thereby alleviating immune-inflammatory responses and thrombosis.
Animals ; Male ; Smad3 Protein/metabolism* ; Transforming Growth Factor beta1/metabolism* ; Rats, Sprague-Dawley ; Rats ; Signal Transduction/drug effects* ; Hypertension, Pulmonary/genetics* ; Thrombosis/immunology* ; Drugs, Chinese Herbal/administration & dosage* ; Humans ; Apoptosis/drug effects*

Glutamine provides carbon and nitrogen to support the proliferation of cancer cells. However, the precise reason why cancer cells are particularly dependent on glutamine remains unclear. In this study, we report that glutamine modulates the tumor suppressor F-box and WD repeat domain-containing 7 (FBW7) to promote cancer cell proliferation and survival. Specifically, lysine 604 (K604) in the sixth of the 7 substrate-recruiting WD repeats of FBW7 undergoes glutaminylation (Gln-K604) by glutaminyl tRNA synthetase. Gln-K604 inhibits SCFFBW7-mediated degradation of c-Myc and Mcl-1, enhances glutamine utilization, and stimulates nucleotide and DNA biosynthesis through the activation of c-Myc. Additionally, Gln-K604 promotes resistance to apoptosis by activating Mcl-1. In contrast, SIRT1 deglutaminylates Gln-K604, thereby reversing its effects. Cancer cells lacking Gln-K604 exhibit overexpression of c-Myc and Mcl-1 and display resistance to chemotherapy-induced apoptosis. Silencing both c-MYC and MCL-1 in these cells sensitizes them to chemotherapy. These findings indicate that the glutamine-mediated signal via Gln-K604 is a key driver of cancer progression and suggest potential strategies for targeted cancer therapies based on varying Gln-K604 status.
Glutamine/metabolism* ; Myeloid Cell Leukemia Sequence 1 Protein/genetics* ; Humans ; Proto-Oncogene Proteins c-myc/genetics* ; Cell Proliferation ; Signal Transduction ; Neoplasms/pathology* ; F-Box-WD Repeat-Containing Protein 7/genetics* ; Cell Survival ; Cell Line, Tumor ; Apoptosis

Tacrolimus is an immunosuppressant that was clinically used for organ transplantation in the 1990s. In the early 2000s, tacrolimus began to be used to treat pediatric kidney diseases in China. This article reviews the therapeutic effects, clinical dosages, and treatment methods of tacrolimus in the treatment of steroid-resistant, steroid-dependent, frequently relapsing, different pathological types, and monogenic mutation-related childhood nephrotic syndrome. It explores the clinical guiding role of machine learning in tacrolimus treatment for childhood nephrotic syndrome, aiming to provide references for the clinical research and application of tacrolimus in pediatric kidney diseases.

Objective To retrieve,evaluate and summarize the best evidence for blood glucose monitoring and management in patients with diabetes of the exocrine pancreas(DEP),and to provide evidence-based basis for clinical practice.Methods According to the"6S"evidence model,relevant evidence on blood glucose management in patients with DEP was searched from computer decision support systems,guideline networks,professional association websites and databases from top to bottom,covering clinical decision-making,clinical practice guidelines,evidence summaries,systematic reviews,expert consensuses,and randomized controlled trials.The search scope was from the establishment of the databases to June 30,2024.Totally 2 researchers independently evaluated the quality of the included literature,and extracted data and summarized evidence that met the criteria.Results A total of 10 articles were included,including 3 guidelines,1 expert consensus,2 systematic reviews,l clinical decision,1 RCT,and 2 cohort studies.Finally,26 pieces of best evidence were formed from 5 aspects,including blood glucose management team construction and goals,blood glucose monitoring methods,glucose control medication management,lifestyle health education and treatment of hypoglycemia.Conclusion This study summarizes the best evidence for blood glucose monitoring and management in DEP patients,which can provide resource preparation for clinical translation and a basis for medical staff to carry out blood glucose management in DEP patients.

Objective:To explore the risk factors for complications of facial autologous fat transplantation.Methods:A total of 51 female patients (case group) with moderate to severe complications following facial autologous fat transplantation at the Fourth Medical Center of Chinese PLA General Hospital from November 2016 to October 2022 were included in this retrospective study. The median age was 31.0 (27.0, 40.0) years. After age and surgical date were matched with ratio of 1∶1, a total of 51 female patients who received autologous fat transplants at several official medical facilities and experienced no complications within a year after the procedure made up the control group. The median age of the control group was 32.0 (26.0, 41.0) years. The clinical characteristics of the two groups were compared, and the factors for complications of facial autologous fat transplantation were examined using a multivariate logistic regression model.Results:In the case group, complications included facial artery embolism (7 cases), ophthalmic artery embolism (19 cases), infection (19 cases), and fat necrosis (6 cases), with 26 severe and 25 moderate cases. No significant differences were found between the two groups in age, body mass index (BMI), marital status, history of hypertension, infectious diseases, allergies, smoking, or alcohol consumption (all P>0.05). However, significant differences were observed in a history of facial surgery, perimenstrual phase, surgical site, and fat donor site (all P<0.05). Multivariate logistic regression analysis revealed that a history of facial surgery ( OR=17.289, 95% CI: 4.851-61.616, P<0.001) and the surgical site being a clinic/outpatient department (compared to a hospital, OR=7.708, 95% CI: 2.482-23.939, P<0.001) were risk factors for postoperative complications after facial autologous fat transplantation. Conclusion:A history of facial surgery and the surgical site being a clinic/outpatient department (compared to a hospital) are risk factors for complications of facial autologous fat transplantation.

Objective To investigate the application value of arterial spin labeling(ASL)perfusion weighted imaging in the early diagnosis of Alzheimer's disease(AD).Methods A total of 60 patients with different ages(control group)and 60 AD patients with different degrees(patient group)who underwent cerebral ASL perfusion weighted imaging examination were selected.At the work-station,the region of interest(ROI)of the hippocampus and limbic system brain structures were delineated and the cerebral blood flow(CBF)value of the ROI were measured.The CBF values of the brain structures of the control group at different ages and the corresponding ROI of the control group and the patient group were compared.Results There were statistically significant differences in the CBF values of the cingulate gyrus,parahippocampal gyrus,hippocampus,orbital part of frontal lobe,temporal pole of temporal lobe,insula lobe and amygdaloid body of the limbic system among the different ages in the control group(P＜0.05),and the CBF value of brain structures gradually decreased with the increase of age.There were statistically significant differences in CBF values of ROI between the control group and mild,moderate,severe patient group(P＜0.05),and the CBF value of brain structures gradually decreased with the severity of AD condition.Conclusion The CBF value of ASL perfusion weighted imaging can objectively reflect the blood flow changes and functional status of the hippocampus and limbic system brain structures in elderly people of different ages and AD patients of different degrees,and has important value in the early diagnosis of AD patients.