Analysis of neural stem cells' movements is one of the important parts in the fields of cellular and biological research. The main difficulty existing in cells' movement study is whether the cells tracking system can simultaneously track and analyze thousands of neural stem cells (NSCs) automatically. We present a novel cells' tracking algorithm which is based on segmentation and data association in this paper, aiming to improve the tracking accuracy further in high density NSCs' image. Firstly, we adopted different methods of segmentation base on the characteristics of the two cell image sequences in our experiment. Then we formed a data association and constituted a coefficient matrix by all cells between two adjacent frames according to topological constraints. Finally we applied The Hungarian algorithm to implement inter-cells matching optimally. Cells' tracking can be achieved according to this model from the second frame to the last one in a sequence. Experimental results showed that this approaching method has higher accuracy compared with that using the topological constraints tracking alone. The final tracking accuracies of average of sequence I and sequence II have been improved 10.17% and 4%, respectively.
Algorithms ; Animals ; Cell Count ; Cell Movement ; Cell Tracking ; statistics & numerical data ; Image Processing, Computer-Assisted ; methods ; Microscopy, Fluorescence ; Models, Theoretical ; Neural Stem Cells ; cytology

OBJECTIVETo investigate the levels and functions of CD4(+)CD25(+) regulatory T cells and specific transcription factor Foxp3 and Th17 cells related cytokine in peripheral blood mononuclear cells (PBMC) and renal tissues, and explore their roles in pathogenesis of Henoch-Schonlein purpura nephropathy (HSPN) in children.

METHODFrom March, 2011 to March, 2013, 30 cases of HSPN children underwent renal biopsy and were treated in Guiyang Children's Hospital were enrolled into this study. Ten healthy children who underwent health check up were enrolled as blood sample control group. The normal kidney tissue specimens were taken from 5 children who underwent surgery for urologic disorders were used as renal sample control group. The circulating proportions of CD4(+)CD25(+) regulatory T cells in PBMC of 30 cases of HSPN children and 10 cases of control group were determined by flow cytometry, respectively.Reverse transcription-polymerase chain reaction (RT-PCR) were used to analyze the mRNA expressions of IL-17, IL-1β and Foxp3 in PBMC. The expression of IL-17 and IL-1β in renal tissue of HSPN and control group were measured by immunohistochemistry. CD4(+)CD25(+) regulatory T cells, Foxp3, IL-17, IL-1β expression were analyzed and compared in HSPN group and control groups respectively.

RESULTThirty cases of HSPN pathological classification were as follows: type I was found in 0 case; type II in 9 cases; type III in 16 cases; type IV in 5 cases; type V in 0 case. The circulating proportions of CD4(+)CD25(+)/CD4(+)T cells and the CD4(+)CD25(+)Foxp3(+)Treg/CD4(+)T cells level were (5.84 ± 0.78)%, (1.01 ± 0.46) % in HSPN groups were substantially lower than those in control group. All these two differences had statistical significance (t = 27.200, 33.260, P < 0.05). The mRNA levels of IL-17, IL-1β in HSPN groups (0.86 ± 0.01,0.71 ± 0.01) were higher than those in control group (t = 25.000, 31.840, all P < 0.05). Foxp3 mRNA expression in HSPN groups (0.24 ± 0.02) were significantly lower than those in control group (t = 21.690, P < 0.05). Protein expression of IL-17 and IL-1β in renal tissues of HSPN children (13.31 ± 0.54, 11.56 ± 0.28) were significantly stronger than those in the control group (t = 27.6, 14.0, all P < 0.01). The highest level of protein expression of IL-17 and IL-1β in renal biopsy of HSPN was in type IV (IV>III>II, F = 545.800, 262.500, all P < 0.01).

CONCLUSIONThe disorder of quantity and function of CD4(+)CD25(+) regulatory T cells, and increase in levels of IL-17, IL-1β (cytokine related to Th17 cells) may play important roles in pathogenesis of HSPN in children; increased protein expression of IL-17, IL-1β in renal tissue may contribute to the development of renal pathological damage in HSPN children.

Case-Control Studies ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Forkhead Transcription Factors ; genetics ; metabolism ; Humans ; Interleukin-17 ; genetics ; metabolism ; Interleukin-1beta ; genetics ; metabolism ; Kidney ; metabolism ; pathology ; Male ; Nephritis ; etiology ; immunology ; pathology ; Purpura, Schoenlein-Henoch ; complications ; immunology ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Severity of Illness Index ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology