BACKGROUND:Nowadays, growth factors are commonly used to induce bone marrow mesenchymal stem cells. However, this is a high-cost method with a great amount of growth factors. In addition, the chondrogenic potential of bone marrow mesenchymal stem cells wil decrease significantly with increasing times of culture. OBJECTIVE:To observe the directed differentiation of bone marrow mesenchymal stem cells co-cultured with human synovial fluid. METHODS:Human bone marrow mesenchymal stem cells were isolated and cultured by adherence screening method. The synovial fluid of the knee was aspirated from healthy volunteers by aseptic operation. Passage 3 human bone marrow mesenchymal stem cells were co-cultured with the fol owing media:synovial fluid+complete medium;synovial fluid+bone marrow mesenchymal stem cells+complete medium;bone marrow mesenchymal stem cells+complete medium. The morphology and growth of the cells were observed under an inverted microscope every day. At days 7, 14 and 21 of induction, toluidine blue staining and immunocytochemical staining were performed.
RESULTS AND CONCLUSION:After co-culture with human synovial fluid, human bone marrow mesenchymal stem cells proliferated slowly, and varied from fusiform to oval or polygonal;toluidine blue and col agen II staining were positive. These findings indicate that the synovial fluid has a positive role in the chondrogenic differentiation of bone marrow mesenchymal stem cells. The synovial fluid may contain substances that promote the chondrogenic differentiation of bone marrow mesenchymal stem cells.

Objective To explored the effects of fentanyl on cell proliferation of H1299 cells, Methods After treating H1299 cells with different concentrations of fentanyl (0.001, 0.010, 0.100, 1.000 μM) for 12, 24, 48, 72 h, cell viability was detected by CCK-8 assay; the rate of cell apoptosis was determined by DAPI staining; the expression levels of Bax, Bcl-2, p-AKT and AKT protein were measured by Western blotting; Caspase-3 activity was determined by Caspase-3 activity assay kit. Results Compared with the control group, fentanyl obviously inhibited the viability of H1299 cells in a dose and time dependent way. Moreover, treatment with different concentrations of fentanyl(0.001, 0.010, 0.100, 1.000 μM) for 12, 24, 48, 72 h, the apoptosis rate of H1299 cells were significantly increased, The level of Bcl-2 protein reduced the level of Bax protein, and the activity of Caspase-3 in H1299 cells were increased after treatment with fentanyl (0.010, 0.100, 1.000 μM) for 48 h, Furthermore, fentanyl markedly inhibited p-AKT/AKT activity of H1299 cells. Conclusions Fentanyl can inhibit cell proliferation and promote cell apoptosis of human lung cancer, and its mechanism may be related to inhibition of AKT activation ,

Objective This study aims to investigate the sex difference in the hippocampus and parahippocampus in patients with bipolar disorder. Methods We acquired T1－weighted structural MRI from 133 bipolar type I patients (60 males) and 144 normal controls (81 males). The General Linear Model was used to examine the relationship between sex and brain volumes of the hippocampus and parahippocampus, with age and intracranial volume as covariates. Results Patients showed significantly smaller volumes of the bilateral hippocampus and parahippocampus (P＜0.01). There were sex－by－diagnosis interactions in the left parahippocampus gyrus (F=6.534, P=0.044). Male patients had significant smaller volumes of the left parahippocampus gyrus compared to the male normal controls (P＜0.001) whereas the volumes were not significantly different between female patients and female normal controls (P＞0.05). Conclusion The results suggest sex difference in the left parahippocampus gyrus volume in patients with bipolar type I disorder, which deserves further investigation in the future bipolar imaging researches.

Objective:To explore the effect and mechanism of fentanyl on promoting the stemness of breast cancer cell.Methods:From January 2018 to October 2019, human breast cancer cell line BT549 was used as the in vitro research object. Breast cancer cell BT549 was pretreated with 0.01 and 0.10 μmol/L fentanyl. Sphere formation assay and colony formation assay were performed to investigate the role of fentanyl on breast cancer cell stemness. Fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the mRNA level of stemness-related transcription factors gender-determining area Y box protein 2 (Sox2), octamer binding transcription factor 4 (Oct4) and Nanog. Western blotting assay was performed to determine the level of Wnt3a/β-catenin pathway-related markers Wnt3a, phosphorylated glycogen synthase kinase-3β (p-GSK-3β), glycogenase kinase-3β (GSK-3β) and β-catenin. After down-regulating Wnt3a, western blotting assay and sphere formation assay were performed.Results:The sphere diameter, colony formation rate and the expression of Sox2 mRNA, Oct4 mRNA, Nanog mRNA, Wnt3a, p-GSK-3β, GSK-3β and β-catenin in 0.01 and 0.10 μmol/L fentanyl-treated breast cancer cell were significant higher than those in blank control: (131.22 ± 1.06) and (636.37 ± 0.02) μm vs. (72.68 ± 0.13) μm, (41.33 ± 0.03)% and (60.58 ± 1.08)% vs. (20.93 ± 0.15)%, 2.25 ± 0.20 and 3.82 ± 0.84 vs. 1.00, 1.87 ± 1.06 and 3.35 ± 0.04 vs. 1.00, 2.85 ± 0.03 and 4.36 ± 0.50 vs. 1.00, 1.82 ± 0.03 and 2.57 ± 0.42 vs. 1.00, 2.04 ± 0.13 and 2.81 ± 0.05 vs. 1.00, 1.62 ± 0.17 and 2.93 ± 0.06 vs. 1.00, 2.15 ± 0.02 and 3.54 ± 0.21 vs. 1.00, the indexes in 0.10 μmol/L fentanyl-treated breast cancer cell were significantly higher than those in 0.01 μmol/L fentanyl-treated breast cancer cell, and there were statistical differences ( P<0.01). After down-regulating Wnt3a, the expressions of p-GSK-3β, GSK-3β, β-catenin, Sox2, Oct4 and sphere diameter were significantly lower than those in blank control: 0.12 ± 0.05 vs. 1.00, 0.53 ± 0.06 vs. 1.00 and 0.24 ± 0.21 vs. 1.00, 0.28 ± 0.10 vs. 1.00 and 0.06 ± 0.01 vs. 1.00, (18.14 ± 0.30) μm vs. (74.32 ± 0.12) μm, and there were statistical differences ( P<0.01). Conclusions:Fentanyl promotes breast cancer cell stemness by Wnt3a/β-catenin signaling pathway.

Objective: To investigate the development status and risk factors of infants and toddlers in rural China. Methods: In this cohort study, 603 infants (6-12 months of age, Phase Ⅰ) in the rural areas of QinLing-Bashan (Qin-Ba) in Shaanxi were recruited in the control group that received no intervention from April 2013 to October 2015. Three follow-up visits were performed every six months (Phase Ⅱ(12-18 months of age), Phase Ⅲ (18-24 months of age) and Phase Ⅳ(24-30 months of age)). In all the 4 phases (Ⅰ-Ⅳ), general data of the children and the families were collected by questionnaires, early childhood growth and development were assessed by door to door visits, children's hemoglobin levels were determined by laboratory tests, and the cognitive and motor development screening was conducted by the Bayley Scales of Infant and Toddler Development. Logistic regression was used to analyze the risk factors affecting the development of infants and toddlers in rural areas and the data were analyzed in terms of risk factors from infants, guardians and family. Results: Phase Ⅱ, Phase Ⅲ and Phase Ⅳ survey recruited 497, 483 and 486 participants respectively. The incidences of cognitive impairment (mental development scores<80) in rural areas of southern Shaanxi were 13.4% (81/603) in Phase Ⅰ(6-12 months), 20.1%(100/497) in PhaseⅡ(12-18 months), 42.9% (207/483) in Phase Ⅲ(18-24 months) and 50.4%(245/486) in Phase Ⅳ(24-30 months) respectively, which showed a significant increase with age (χ²=233.40, P<0.01); the incidences of psychomotor impairment (psychomotor development scores<80) of Phase Ⅰ, Phase Ⅱ, Phase Ⅲ and Phase Ⅳ were 25.0% (151/603), 26.8% (133/497), 8.3% (40/483) and 11.9% (58/486), which showed a significant decrease with age (χ²=87.08, P<0.01). Multivariate logistic regression analysis showed that the leading risk factor of the cognitive development of 24-30-month-old children was the mothers' poor education background (≤9 years of school education) (OR=2.56, P<0.01), and the main risk factors affecting psychomotor development were the mothers' poor education background (≤9 years of school education) (OR=2.64, P<0.05) and growth retardation (OR=2.95, P=0.07). Conclusions The early childhood development (especially cognitive development) in the rural areas of Qin-Ba in Shaanxi of China is not optimistic. More attention should be paid to the early childhood development in rural China, especially to the development of children from the mothers with poor education background.

Objective:To study the logistic regression model and Chi-square automatic interaction detection decision tree model in the prediction of the recurrence of acute pancreatitis (AP).Methods:Clinical data of 364 patients with AP admitted to the Affiliated Hospital of Jiangnan University from June 2021 to June 2022 were retrospectively analyzed, including 219 males and 145 females, aged 53 (19-91) years. The patients were divided into the recurrence group ( n=63), those who experienced a second or more episodes of AP, and the initial group ( n=301), those who were diagnosed of AP for the first time. Univariate and multivariate logistic regression analyses were performed to identify the factors associated with recurrence of AP, and the decision tree model was used to analyze those factors. Receiver operating characteristic (ROC) curve were plotted to analyze the predictive performance of the two models. Results:Multivariate logistic regression analysis showed that age ( OR=0.969, 95% CI: 0.949-0.990, P=0.004), body mass index ( OR=1.142, 95% CI: 1.059-1.232, P=0.001), and hyperlipidemia ( OR=3.034, 95% CI: 1.543-5.964, P=0.001) were independent factors influencing the recurrence of AP. The accuracy of the model in predicting recurrence was 83.2% (303/364). The decision tree model showed that hyperlipidemia and body mass index were factors influencing the recurrence of AP, with an accuracy of 82.7% (301/364) in predicting recurrence. The area under the ROC curve was larger in the logistic regression model compared to that in the decision tree model (0.776 vs 0.730, Z=2.02, P=0.043). Conclusion:The logistic regression model and the Chi-square automatic interaction detection decision tree model can help predict the recurrence of AP. It is recommended to combine the two models to better guide clinical practice.

BACKGROUND:How to improve the hematopoietic microenvironment is a hot topic in the treatment of aplastic anemia. OBJECTIVE:To explore the action mechanism of Angelica sinensis polysaccharide in the treatment of aplastic anemia by combining GEO sequencing analysis,network pharmacology,and experimental validation. METHODS:Aplastic anemia-related differentially expressed genes were obtained from GEO database,and gene ontology and gene set enrichment analysis were performed.The active components and targets of Angelica sinensis polysaccharide were obtained by combining the literature with PubChem,SwissTargetPrediction,and PharmMapper databases.After the intersection targets were taken,STRING and Cytoscape were used to construct protein-protein interaction network,and gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis was performed.Mouse model of aplastic anemia was established,and the effect and action mechanism of Angelica sinensis polysaccharide on aplastic anemia were verified by blood cell analyzer,flow cytometry,ELISA,immunohistochemistry,immunofluorescence,and western blot assay. RESULTS AND CONCLUSION:(1)A total of 834 differentially expressed genes were screened,which were involved in biological processes such as cell development,hematopoiesis,and myeloid cell differentiation.(2)347 targets related to Angelica sinensis polysaccharide were retrieved and 77 potential therapeutic genes were screened.Among them,the degree values of angiogenic,apoptotic,and immune-related factors such as VEGFA,EGLN1,Bcl-2,interferon-γ,interleukin-2,interleukin-4,and interleukin-6 were significant.(3)KEGG pathway enrichment analysis revealed that the therapeutic targets were mainly enriched in Th17 cell differentiation,NK-related cytotoxicity,cell adhesion factors such as interferon-γ,interleukin-2,and interleukin-4 related signaling pathways.(4)Animal experiments showed that Angelica sinensis polysaccharide significantly improved bone marrow haematopoiesis,increased peripheral blood cell,and bone marrow single nucleated cell counts,and improved the survival rate of mice.Compared with the model group,mice in the Angelica sinensis polysaccharide group showed a significant decrease in the ratio of Th1/Th2 cells(P<0.01),a decrease in the expression level of interferon-γ(P<0.01),an increase in the level of interleukin-4(P<0.05),a significant increase in the level of VEGFA(P<0.01),a significant decrease in EGLN1(P<0.01),a significant decrease in apoptosis rate of bone marrow single nucleated cells and reactive oxygen species level(P<0.01),and a significant increase in cleaved Caspase-3 protein expression(P<0.01),and a significant decrease in Bcl-2/Bax ratio(P<0.01).(5)These findings show that Angelica sinensis polysaccharide can improve hematopoiesis of aplastic anemia mice by regulating aberrant T-cell subsets and promoting angiogenesis to improve hematopoietic microenvironment,and inhibiting apoptosis of bone marrow mononuclear cells.

Objective:To qualitatively analyze the cardiac rehabilitation information need based on KANO model in patients after percutaneous coronary intervention (PCI) and to obtain the satisfaction and importance of every need so as to provide a basis for optimizing clinical rehabilitation health guidance.Methods:From May 2019, this study selected 144 patients with selective PCI as subjects by convenience sampling. The KANO two-factor questionnaire was designed based on 10 aspects of needs including the basic knowledge, nutrition, exercises, medications, works and professions, pressure and psychology, social concern, crisis and safety, diagnosis and therapy, risk factor management of cardiac rehabilitation information need, to gain the need class and draw the need satisfaction-importance magic quadrant.Results:Among 51 items in the cardiac rehabilitation information need questionnaire, there were 5 of them (9.8%) with essential attribute, 14 of them (27.5%) with expected attribute, 8 of them (15.7%) with glamour attribute and 24 of them (47.1%) with no difference attribute. The satisfaction-importance matrix diagram showed that 4 aspects of need were in the area for improvement and 20 of them in the area for the secondary improvement among cardiac rehabilitation information needs after PCI.Conclusions:The need importance matrix directly reflects the patient satisfaction and indirectly reflects patients' need importance.

In order to characterize the immunogenicity and immunoprotection of the Staphylococcus aureus (S. aureus) surface protein Clumping factor A (ClfA), we amplified clfa genes from S. aureus Newman strain, Wood46 strain and HLJ23-1. The clfa gene from Newman strain was subsequently inserted into pQE-30 vector and the recombinant plasmid was transformed into Escherichia coli strain M15 (pREP4). The recombinant ClfA protein was expressed and purified. Then, we immunized mice with the purified recombinant protein. The antibody level and the concentration of cytokines were measured by enzyme-linked immunosorbent assay. Finally, immunized mice were challenged with S. aureus Newman, Wood46 and HLJ23-1. These results suggested that clfa gene sequences were highly conserved, and the recombinant ClfA was expressed correctly with good antigenicity. The antibody titer and the concentration of cytokines in the immunized groups increased significantly (P < 0.05) compared with control, and the mice in the immunized groups were protected against the challenge strains to some extent. These results showed that the ClfA had high immunogenicity and immunoprotective potential.
Animals ; Coagulase ; genetics ; immunology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Immunization ; Mice ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Staphylococcus aureus ; metabolism ; pathogenicity

Objective: To compare the clinical effects between minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) assisted by Microendoscopic discectomy (MED) and Quadrant for the treatment of degenerative lumbar spinal stenosis (DLSS). Methods: All of 59 patients suffered from DLSS treated surgically from May 2015 to October 2017 were reviewed. According to the surgery method, all cases were divided into MED group (27 cases) and Quadrant channel group (32 cases). All patients were followed up for an average of 18.5 months (11-29 months). Comparison was made on the operative time, intraoperative blood loss, postoperative drainage, postoperative time in bed, postoperative creatine kinase (CK), fusion rate and the degree of muscle fibrosis shown in MRI, as well as visual analogue scale (VAS)score and Oswestry dysfunction index (ODI) score in two groups. Results: The duration of operation in MED group was significantly longer than that in Quadrant group (161.7±22.4 min vs. 145.6±19.4 min, t=4.541, P<0. 01), but less intraoperative blood loss (138.1±26.9 ml vs. 155.6±21.5 ml, t=-2.724, P< 0. 01) and shorter time in bed after surgery (2.3±0.7 d vs. 3.5±1.1 d, t=-4.564, P<0.01). Compared with Quadrant group, CK levels were risen slightly on the first and third postoperative day (P<0.01). CK on the fifth postoperative day in both groups returned to normal. VAS score of lower back pain in MED group was lower than that in Quadrant group on the third day and twelfth month after operation (P<0.05). After 3 months, there was no significant difference in VAS score and ODI between the two groups (P>0.05). ODI was lower in MED group than that in Quadrant group after 6 months and 12 months.The fusion rate was88.9%(24/27) in MED group and 93.8%(30/32) in Quadrant channel group. There was no statistical difference in fusion rate of two groups. 10 patients in MED group and 12 patients in Quadrant group underwent MRI examination of lumbar spine one year after operation. The ratio of postoperative and preoperative atrophy of multiplex muscle area was measured. Muscle atrophy of lower back muscle was lighter in MED group (0.12±0.05 vs. 0.22±0.04, t=-2.428, P<0.05). For intraoperative and postoperative complications, 1 case of dural sac rupture occurred in both groups. Gelatin sponge immediately with fibrin glue was used for plugging up, no postoperative cerebrospinal fluid leakage was found. In Quadrant channel group, 1 case had less blood supply of skin incision edges and epidermal necrosis while the other case had fat liquefaction. Conclusion Compared with the aid of Quadrant, MIS-TLIF assisted with MED had less blood loss, less trauma and faster recovery and could reduce the incidence of postoperative incision complication.