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 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 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 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.

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.

Objective To explore the predictive value of CT radiomics and morphological features for the prognosis and survival in non-small cell lung cancer(NSCLC)patients.Methods The clinic data of 300 NSCLC patients(300 lesions)were downloaded from the Cancer Imaging Archive,with 210 randomly selected as the training set and 90 as the test set.According to the prognosis and survival,the patients were divided into two groups with survival period≤3 and＞3 years.3D Slicer software was used to delineate the regions of interest layer by layer in CT images,and the radiomics features were extracted from each region of interest.Both t-test and least absolute shrinkage and selection operator were utilized for radiomics feature screening.Three types of prediction models,namely radiomics model,morphological model and combined model,were constructed with Logistic regression,whose performances were evaluated using the receiver operating characteristic(ROC)curve.Results The differences in radiomics labels and mediastinal lymph node metastasis between the training set and the test set were statistically significant.For radiomics model,morphological model and combined model,the area under the ROC curve was 0.784(95%CI:0.722-0.847),0.734(95%CI:0.664-0.804)and 0.748(95%CI:0.680-0.815)in the training set,and 0.737(95%CI:0.630-0.844),0.665(95%CI:0.554-0.777)and 0.687(95%CI:0.578-0.797)in the test set,which demonstrated that radiomics model had the best diagnostic performance.Conclusion The CT radiomics model can effectively predict the prognosis and survival in NSCLC patients.

ObjectiveTo observe the clinical efficacy of modified Yigongsan combined with multi-enzyme tablets and bifidobacterium triple live powder on infantile anorexia with spleen-stomach Qi deficiency syndrome. MethodA total of 112 infantile patients anorexia with spleen-stomach Qi deficiency syndrome treated at Hebei Provincial Hospital of Traditional Chinese Medicine from January 2022 to June 2023 were enrolled and divided into a control group and an observation group， with 56 cases in each group， according to a random number table. Children in the control group were treated with multi-enzyme tablets and Bifidobacterium triple live powder， while those in the observation group were treated with modified Yigongsan in addition to the treatment in the control group. During the study， one case dropped out in the control group and two cases dropped out in the observation group. The clinical efficacy of the two groups of children was compared， including changes in traditional Chinese medicine （TCM） syndrome scores （main symptoms， secondary symptoms， tongue， and pulse）， time to restore normal food intake， and increase in body weight. Changes in calcium， iron， zinc levels， hemoglobin， and albumin levels before and after treatment， as well as changes in gastrointestinal hormones such as gastrin and motilin， vasoactive intestinal peptide， somatostatin， neuropeptide Y， orexin， and leptin， were observed. The occurrence of adverse reactions in the two groups of children during the study was also recorded. ResultThe total effective rate of children in the control group after treatment was 85.19% （46/54）， while that in the observation group was 98.15% （53/54） （χ² =5.939， P<0.05）. Compared with the control group， the time for food intake to return to normal in the observation group was shorter， and the increase in body weight was greater （P<0.05）. Compared with the results before treatment， the TCM syndrome scores （main symptoms， secondary symptoms， tongue， and pulse） in both groups of children significantly decreased， while the levels of calcium， iron， zinc， hemoglobin， albumin， gastrin， motilin， neuropeptide Y， and orexin increased， and the levels of vasoactive intestinal peptide， somatostatin， and leptin decreased （P< 0.01）. Compared with the control group after treatment， the improvement in the above indicators in the observation group was more significant （P<0.01）. The incidence of adverse reactions in the two groups of children during the treatment period was similar， and the difference was not statistically significant. ConclusionModified Yigongsan combined with multi-enzyme tablets and Bifidobacterium triple live powder is highly effective in treating infantile anorexia （spleen-stomach Qi deficiency syndrome）. After treatment， symptoms of the children were improved，appetite and food intake increased， gastrointestinal function was improved， body weight increased， and adverse reactions were few， indicating that the treatment was safe and reliable.