[Objective]To study the effects of verapamil on the immediate-early gene(IEGs)expression of bone marrow mesenchymal stimulated by mechanical strain,and deduce the effects of calcium ion channel on the early responses of children bone marrow mesenchymal stem cells(MSCs)to mechanical strain in vitro.[Method]MSCs were isolated and cultured.The 3-6th generational MSCs were stimulated in different time by mechanical and(or)verapamil(20?mol/l).Flow cytometry was applied to measure intracellular free Ca2+ at once.Early-response genes /proteins(c-fos,c-jun and c-myc)were examined by RT-PCR and immunocytochemical stain.[Result]The application of mechanical strain increased intracellular c-fos,c-jun and c-myc mRNA /protein levels.The resulting effect is partially inhibited before MSCs were preincubated with verapamil(P

Female ; Humans ; Neuroectodermal Tumors, Primitive ; pathology ; Ovary ; pathology

Objective　To explore the application of time series models based on meteorological factors in the population density of Aedes albopictus in Hainan Province, and to provide a scientific basis for the prevention and control of dengue fever in Hainan Province. Methods　The density of Aedes albopictus in different habitats in 18 cities and counties of Hainan Province from 2017 to 2022 was monitored monthly using the double-mosquito net trapping index and the Breteau index. Mann-Kendall trend test was used to analyze the temporal trends of the two density indices; Spearman's correlation analysis was employed to assess the correlation strength between each meteorological factor and the two indexes, eliminating unrelated variables, and further selecting the final variables through the full-subset regression method. Three time-series models were constructed for the two density indices, with root mean square error (RMSE), mean absolute error (MAE), and other accuracy metrics used to determine the optimal model; predictions for the density indices for 2023-2024 were made. All statistical analyses were performed in R (4.3.1). Results　The net trapping and Breteau indices showed an overall decrease over the years (Z-values of Mann-Kendall trend test were -6.15 and -4.03, respectively, and P<0.05). The meteorological factor most strongly associated with the trap index was the monthly average minimum temperature; monthly mean minimum temperature and monthly mean relative humidity were strongly correlated with the Breteau index. Based on various evaluation indicators, the multivariate time series model demonstrated the highest accuracy. The study predicts one to two peaks in both the trap index and Breteau Index for the years 2023 and 2024, with peak periods occurring between June to September and May to September, respectively. The predicted value for 2023 aligns with the measured value, demonstrating outstanding predictive accuracy. Conclusions　This study has introduced meteorological factors into the seasonal time series model, allowing for more accurate predictions of the density of Aedes albopictus in Hainan Province from 2023 to 2024, providing a model framework for the prevention and control of dengue fever in Hainan Province.

BACKGROUND: It has been reported that in China, human bone marrow mesenchymal stem ceils are mostly harvested from adults. Studies on bone marrow mesenchymal stem cells in children are few.OBJECTIVE: To isolate and expand bone marrow mesenchymal stem cells from children, and to analyze the biological characteristics of bone marrow mesenchymal stem cells and their potential of differentiating into osteoblasts, adipocytes and neural like cells.DESIGN: Observational comparative study.SETTING: Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Experiments were performed at the Laboratory of Department of Orthopaedics of Wuhan Tongji Hospital from March to September 2006. Bone marrow mesenchymal stem cells were collected from one boy patient and two girl patients aged 5-8 years, who received pelvis osteotomy for dysplasia of the hip joint. The experimental procedures were approved by the Hospital Ethics Committee and family members of all children patients singed the informed consent.Dexamethasone, vitamin C, β-sodium glycerophosphate, 3-1sobutyl-1-methylxanthine, insulin, indometacin and butylated hydroxyanisole were bought from Sigma Company. Dimethyl sulphoxide was purchased from Amersco Company.METHODS: Bone marrow mesenchymal stem cells were cultured from mononuclear cells isolated over a Percoll gradient.Bone marrow mesenchymal stem cells were observed under an inverted phase contrast microscope. Bone marrow mesenchymal stem cells could differentiate into osteoblasts, adipocytes and neural like cells with osteoblast inductor (β-sodium glycerophosphate, dexamethasone, vitamin C), lipoblast inductor (dexamethasone, 3-isobutyl-1-methylxanthine,bovine insulin, indometacin) and serum-free medium inductor (dimethyl sulphoxide, butylated hydroxyanisole) respectively.Osteoblast marker (alkaline phosphatase, osteocalcin mRNA, calcium node), adipocyte marker (lipid droplet, PPAR γ-2mRNA) and neural ceil-like marker (nissl body, neuron specific enolase, neurofilament protein) were respectively determined by the immunohistochemical method, polymerase chain reaction and immunocytochemical method.MAIN OUTCOME MEASURES: ①Appearance and proliferation of bone marrow mesenchymal stem ceils from children,and ②determination results of osteoblast, adipocyte and neural cell markers.RESULTS: ①Children bone marrow mesenchymal stem cells could easily adhere to the wall, appeared fusiform, had high reproductive activity and arranged vortically after fusing. ②Appearance of bone marrow mesenchymal stem cells changed after receiving inductor. Osteoblast marker, adipocyte marker and neural cell-like marker were positive after chemical staining, polumerase chain reaction and immunocyte staining.CONCLUSION: Children bone marrow mesenchymai stem cells show stable proliferation, passage and multi-direction differentiation towards osteoblasts, adipocytes and neural like cells.

Objective To investigate the changes in Th17 cells and CD4+CD25+regulatory T lym-phocytes ( Treg) as well as transcription factors and cytokines relating to them in children with Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (HLH) and to analyze their role and clinical significance. Methods Thirty-two children with newly diagnosed EBV-associated HLH in the Hematology/Oncology Department of Zhengzhou Children′s Hospital from January 2012 to December 2016 were enrolled in this study. Thirty healthy children taking physical examination in the same hospital in the corresponding period were recruited as controls. Percentages of Th17 and Treg cells in peripheral blood T lymphocytes were detected by flow cytometry. Expression of RORγt and Foxp3 at mRNA level in peripheral blood mononuclear cells was detected by real-time PCR. Levels of IL-6, IL-17, IL-10 and TGF-β1 in serum samples were measured by ELISA. Results Compared with the control group, the EBV-associated HLH group showed in-creased percentage of Th17 cells [(1. 09±0. 43)％ vs (0. 39±0. 19)％, P<0. 05] and enhanced expres-sion of RORγt at mRNA level [(1. 41±0. 37) vs (0. 67±0. 13), P<0. 05], but decreased percentage of Treg cells [(3. 66±1. 13)％ vs (6. 80±1. 15)％, P<0. 05] and inhibited expression of Foxp3 at mRNA level [(15. 97±5. 11) vs (30. 23±4. 95), P<0. 05]. All of the above mentioned changes were reversed af-ter treatment (P<0. 05). Serum levels of IL-6 and IL-17 of EBV-associated HLH group were higher than those of control group, while serum levels of IL-10 and TGF-β1 were lower (P<0. 05). Conclusion Im-balanced Th17/Treg cells might play an important role in the pathogenesis of EBV-associated HLH. Cyto-kines relating to the maintenance of Th17/Treg cell balance could be used as indicators of disease develop-ment.

OBJECTIVETo investigate the changes and roles of follicular regulatory T cells (Tfr) and follicular T helper cells (Tfh) in the pathogenesis of children immune thrombocytopenia (ITP).

METHODS32 untreated ITP patients, as well as 20 healthy controls were enrolled in this study. The proportion of circulating Tfr and Tfh cells were determined by flow cytometry; real-time PCR was performed to detect the expression of transcription factors and regulatory factors of Bcl-6, c-Maf, Blimp-1 and PD-1 mRNA; ELISA was used to detect plasma concentration of IL-2, IL-6, IL-10 and IL-21.

RESULTS(1)The proportion of Tfh cells were significantly higher (P<0.05), while the Tfr cells and the ratio of tfr/Tfh cells in ITP patients were significantly lower than that in health controls (P<0.05); (2)Correlation analysis showed that the Tfr cells and the ratio of Tfr/Tfh were positively correlated with the platelet counts and negatively with the levels of PA-IgG, while the proportion of Tfh cells was positively correlated with the levels of PA-IgG and negatively with the platelet counts in peripheral blood; (3)Transcription levels of Bcl-6 and c-Maf mRNA in CD4(+) T lymphocytes cells were significantly elevated, the Blimp-1 mRNA in CD4(+) cells and PD-1 mRNA levels of Treg were lower in ITP patients in comparison with healthy controls; (4)The higher Plasma concentration of IL-21, and lower concentration of IL-2 were found in ITP patients.

CONCLUSION(1)The lower proportion of Tfr cells and higher proportion of Tfh cells, as well as the abnormal ratio of Tfr/Tfh might account for the decreased platelet counts to be further involved in the immunological pathogenesis of children ITP; (2)The changes of plasma cytokines IL-2, IL-21 in microenvironment and the over-expression of Bcl-6 mRNA, c-Maf mRNA and the lower-expression of Blimp-1 mRNA in CD4(+) T cells, and over-expression of PD-1 mRNA in Treg cells might be account for the abnormal ratios of Tfr/Tfh cells in ITP patients.

Cell Movement ; Child ; Humans ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; T-Lymphocytes, Helper-Inducer ; immunology ; T-Lymphocytes, Regulatory ; immunology