Objective To explore in vitro the best time window for using sinusoidal electromagnetic fields to promote the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).Methods BMSCs were isolated and cultured from 4-week-old Sprague-Dawley rats (male and female,80-120g).The BMSCs (from passage 3) were exposed 0,1,4 or 8h/d for 7d,14,or 28d,respectively,to 15Hz sinusoidal electromagnetic fields with a maximum amplitude of lmT.Those exposed 0h/d served as the control.The relative expressions of runt related gene-2 (RUNX2),bone sialoprotein (BSP) and osteopontin (OPN) were determined using real-time,quantitative reverse transcription-polymerase chain reactions (RT-PCRs).The level of RUNX2 protein was determined by Western blotting after 14d.Alizarin red staining was used to compare calcium distribution in each group.Results Obvious promotion of differentiation to osteoblasts was observed after 7 days of exposure to the15 Hz sinusoidal electromagnetic fields,most obviously manifested by an outstanding increase of the early osteogenic index RUNX2 in those exposed 4h/d.After 14 days of intervention,the 1h/d exposure showed to be most effective,especially in inducing the changes of the late osteogenic index OPN.The trends of changes in RUNX2 protein were similar in all groups.After stimulating 1h/d for 14 and 28days,calcium deposition increased to the greatest extent.Conclusions Exposure to sinusoidal electromagnetic fields induces osteogenic differentiation to osteoblasts in rat BMSCs in vitro.There is an apparent window effect.The best results are observed with more days of exposure and shorter exposure time (1h) every day.

OBJECTIVE:To optimize the extraction technology of glucosinolates from Uighur medicine Brassica rapa L.. METHODS:Ethanol refluxing method was adopted to extract glucosinolates from Uighur medicine B. rapa L.. With the comprehen-sive score of glucosinolates and dry extract yield as the index,L9(34)orthogonal test based on the single factor test was adopted to observe the effects of ethanol volume fraction,extraction time and material-liquid ratio on the extraction technology of glucosino-lates from B. rapa L.,and verification test was conducted. RESULTS:The optimal extraction technology was to add 95% ethanol 8 times as much as the amount of the herbs,twice for 1.0 h reflux extractions. The extraction amount of glucosinolates and dry ex-tract yield were 7.36 mg/g and 25.29% at average,respectively.The comprehensive score of RSD was 0.52%(n=3). CONCLU-SIONS:The optimal extraction technology is stable and feasible and can be used for the extraction of glucosinolates from B. rapa L..

Aim To investigate the anti-angiogenesis and anti-xenograftes of UA in zebrafish larvaes. Meth-ods 24 hour post-fertilization ( hpf ) TG zebrafish was treated with different concentration of UA for 24h, and the number of intersegmental vessels( IVS) were detec-ted under fluorescent microscope respectively;then the models of AB/TG zebrafish xenografts were established by be micro-injected with SMMC-7721 or HT-29 cell at 48hpf respectively, and after cocultured with UA for 48h, optical density (OD) of the SMMC-7721 cell and subintestinal veins ( SIVs) induced by HT-29 were e-valuated under confocal microscope. Results ISV as-say showed that UA could cause IVS missing or disap-perance, inhibition ratio reaching 20. 25% and 26. 65%. UA blocked the spread of SMMC-7721 cells in zebrafish and OD value,and inhibition ratios at three concentrations were 38. 01%, 54. 69%, 61. 88%, re-spectively; in another SIVs assay induced by xeno-grafts, UA at concentration 10 and 15mg·L-1 showed that SIVs were inhibited (P<0. 01) obviously. Con-clusion UA could inhibit the angiogenesis and the growth of SMMC-7721/HT-29 xenografts,and the anti-tumor mechanism may be related with VEGFR2 expres-sion.

OBJECTIVE: To explore the change in ambulatory blood pressure monitoring (ABPM) value and the sympathetic nervous system (SNS) level in children with primary nephrotic syndrome(PNS) and their relationship. METHODS: ABPM and casual blood pressure(CBP) were tested in 114 children with PNS and 12 normal children as a control group. The 24-h urine noradrenaline(NA), adrenaline(A) and dopamine(DA) content were detected through high-performance liquid chromatography with electrochemical luminescence and the correlation with ABP was analyzed. RESULTS: Among 114 children with PNS, 101 had elevated blood pressure (88.6%), 45 showed high incidence of masked hypertension (39.5%), and 80 non-dipper blood pressure (70.2%). Systolic blood pressure level and blood pressure load were greater than diastolic blood pressure. NA, A, and DA levels of the PNS group were significantly higher than those of the control group, while those of the elevated blood pressure group were significantly higher than those of the normal blood pressure group in PNS children. SNS levels were positively correlated with blood pressure levels and blood pressure load, and negatively correlated with night BP decreasing rates. CONCLUSION Children with PNS have high incidence of hypertension with large proportion of masked hypertension and non-dipper blood pressure. Severe masked hypertension classification should be set up. In PNS children, SNS activity is elevated that might evaluate the blood pressure level and decrease blood pressure circadian rhythm.
Adolescent ; Blood Pressure ; physiology ; Blood Pressure Monitoring, Ambulatory ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Hypertension ; diagnosis ; etiology ; Male ; Nephrotic Syndrome ; complications ; physiopathology ; Sympathetic Nervous System ; physiopathology

Ameloblastoma is a benign tumor characterized by locally invasive phenotypes,leading to facial bone destruction and a high recurrence rate.However,the mechanisms governing tumor initiation and recurrence are poorly understood.Here,we uncovered cellular landscapes and mechanisms that underlie tumor recurrence in ameloblastoma at single-cell resolution.Our results revealed that ameloblastoma exhibits five tumor subpopulations varying with respect to immune response(IR),bone remodeling(BR),tooth development(TD),epithelial development(ED),and cell cycle(CC)signatures.Of note,we found that CC ameloblastoma cells were endowed with stemness and contributed to tumor recurrence,which was dominated by the EZH2-mediated program.Targeting EZH2 effectively eliminated CC ameloblastoma cells and inhibited tumor growth in ameloblastoma patient-derived organoids.These data described the tumor subpopulation and clarified the identity,function,and regulatory mechanism of CC ameloblastoma cells,providing a potential therapeutic target for ameloblastoma.

Pleomorphic adenoma (PA) is the most common benign tumour in the salivary gland and has high morphological complexity. However, the origin and intratumoral heterogeneity of PA are largely unknown. Here, we constructed a comprehensive atlas of PA at single-cell resolution and showed that PA exhibited five tumour subpopulations, three recapitulating the epithelial states of the normal parotid gland, and two PA-specific epithelial cell (PASE) populations unique to tumours. Then, six subgroups of PASE cells were identified, which varied in epithelium, bone, immune, metabolism, stemness and cell cycle signatures. Moreover, we revealed that CD36⁺ myoepithelial cells were the tumour-initiating cells (TICs) in PA, and were dominated by the PI3K-AKT pathway. Targeting the PI3K-AKT pathway significantly inhibited CD36⁺ myoepithelial cell-derived tumour spheres and the growth of PA organoids. Our results provide new insights into the diversity and origin of PA, offering an important clinical implication for targeting the PI3K-AKT signalling pathway in PA treatment.
Humans ; Adenoma, Pleomorphic/genetics* ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; Transcriptome ; Myoepithelioma