Objective The aim of this study was to investigate how different concentrations of dextran sulfate sodi-um ( DSS) influence the establishment of mouse model of inflammatory bowel disease ( IBD) and the effect of DSS on the expression of colitis-associated immune factors.Methods The DSS solution in different concentrations (3%, 5%, 7%) were given to male C57BL/6J mice to generate mouse inflammatory bowel disease model.The IBD mice were observed by defecation characteristics, body weight, and survival time.The animals were sacrificed at 6 days after the start of DSS drinking.The general appearance of colons was observed and scored.Moreover, the pathological changes of the colon were examined and analyzed by routine histology.The expression of immune factors in the spleen was detected by real-time PCR.Results The mice in the 3%, 5%, 7% DSS groups developed murine colitis.In addition, the incidence of IBD and mouse mortality rate was directly proportional to the increase of DSS concentration.Furthermore, the higher concentra-tion of DSS induced the expression of proinflammatory factors including TNF-α, IFN-γand IL-17A, but cause a decrease of anti-inflammatory factors such as IL-4, IL-10 and Treg-related transcription factor Foxp3.Conclusions Our data suggest that giving 5%DSS solution to C57BL/6J mouse is appropriate to efficiently establish a murine IBD model.This laid an important foundation for further studies of the pathogenesis of IBD, biological characteristics, and intervention factors.

Bone marrow hematopoietic microenvironment occupies the medullary cavities of bones throughout the skeleton and provides support for hematopoiesis and immune cells development. Bone-resorbing osteoclasts in bone marrow environment are specialized cells derived from the hematopoietic stem cells and play a pivotal role in process termed as bone remodeling that involves break down and build-up of bone. It is only recently that studies have provided a novel basis for understanding potential role of osteoclasts in homeostasis, stress-induced mobilization of hematopoietic progenitors and osteoimmunology. Further exploration on the interaction of osteoclasts with others in bone marrow hematopoietic microenvironment may contribute to future clinical treatments for hematopoietic and bone-related immunologic disorders including cancer. In this review the origin and identification of osteoclasts and regulation of mobilizing hematopoietic stem cells, as well as osteoclasts and osteoimmunity were mainly concerned.
Bone Marrow Cells ; physiology ; Bone Remodeling ; immunology ; Hematopoiesis ; physiology ; Hematopoietic Stem Cells ; cytology ; immunology ; Homeostasis ; immunology ; Humans ; Osteoclasts ; cytology ; immunology ; physiology

This study was aimed to investigate the effect of RPMI 1640 and IMDM on the development of human peripheral blood monocyte-derived dendritic cells. Under the same cytokines and culture conditions, the different medium types were tested, and the morphology of mature and immature dendritic cells was observed by microscopy, the cell phenotype and endocytosis ability were detected by flow cytometry. Furthermore, the immunoregulatory function of various DC was analyzed by mixed lymphocyte reaction (MLR), the expression of cytokine in culture supernatant of MLR system was also analyzed by Bio-plex technology. The results showed that there were no difference in morphology, CD14, CD83 expression and endocytosis ability between IMDM-cultured DC and RPMI-1640 medium-cultured DC, but there was a lower expression of CD1a in IMDM-cultured DC. Moreover, DC cultured with IMDM displayed a significant reduction in stimulating T cell proliferation, and highly expressed IL-6, IL-8 and IL-10, but low expressed IL-12. It is concluded that the different cultural mediums can induce DC with different functions and DC cultured with IMDM may correlated with induction of immune tolerance. The results of this study will provide a new idea for DC clinical application.
Cell Differentiation ; Cells, Cultured ; Culture Media ; pharmacology ; Cytokines ; Dendritic Cells ; cytology ; Flow Cytometry ; Humans ; Leukocytes, Mononuclear ; cytology ; Lymphocyte Culture Test, Mixed ; Monocytes ; cytology

This study was purposed to investigate the effect of mitogen-activated protein kinase (MAPK) pathway on the osteoblast differentiation of mouse mesenchymal stem cells (MSCs), MSCs were isolated from mouse compact bone and serially passaged. After being cultured in osteogenic induction medium, the phosphorylation levels of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 were detected by Western blot. The effects of corresponding pathway inhibitors including PD98059, JNK II and SB203580 on alkaline phosphatase (ALP) and calcium accumulation in the osteoblastic differentiation of MSCs were determined by ALP staining and von kossa staining respectively. The results showed that MAPK pathway including ERK, JNK and p38 was activated in differentiation of MSCs into osteoblasts. ALP activity of MSCs increased in the early phase by addition of PD98059 treatment, whereas ALP activity and calcium accumulation were not observed via JNK II treatment. However, SB203580 strongly inhibited the ALP expression and the calcium accumulation. It is concluded that p38 plays a positive role in the osteogenic differentiation of MSCs, and ERK is probably a negative factor at the early phase of differentiation, but the effect of JNK is not essential.
Animals ; Cell Differentiation ; Cells, Cultured ; Female ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Mitogen-Activated Protein Kinases ; metabolism ; Osteoblasts ; cytology ; Osteogenesis ; Signal Transduction

OBJECTIVETo evaluate magnetic resonance venography (MRV) in diagnosing obstructive interface morphology of Budd-Chiari syndrome(BCS).

METHODSMRV examination was performed on 44 cases of BCS, and the images of obstructive interface morphology of the inferior vena cava were reviewed by two radiologists.

RESULTSIn all 44 cases, there were 37 cases with complete obstruction and 7 with incomplete obstruction. MRV showed 4 cases with membrane with hole of incomplete obstruction. The morphologies MRV demonstrated that the proximal part of the 37 cases with complete obstruction were mainly divided into the cone type (36 cases) and the planum type (1 case). Besides, the type of distal end of obstruction were the cone type (30 cases), the planum type (4 cases) and the irregular type (3 cases). The overall sensitivity, specificity, positive and negative predictive values for the diagnosis of MRV were respectively 100%%, 57.1%, 92.5% and 100% as compared to the DSA.

CONCLUSIONThe examination of MRV is capable of revealing the obstructive interface morphology of the inferior vena cava, especially for the distal end of obstruction. MRV can provide guidelines in interventional treatment of Budd-Chiari syndrome.

Adult ; Aged ; Angiography, Digital Subtraction ; Budd-Chiari Syndrome ; diagnostic imaging ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Vena Cava, Inferior ; diagnostic imaging ; pathology ; Young Adult

This study was aimed to investigate the effect of human umbilical vein endothelial cells (HUVEC) on dendritic cell (DC) development. First, HUVEC were isolated from human umbilical cord by collagenase digestion, and then the morphology, immunophenotypes and functions were identified. Furthermore, the HUVEC were cocultured with CD14(+) monocytes under the cytokine condition for detecting the influence of HUVEC on differentiation of CD14(+) cells to DC. The phenotype of dendritic cells derived from CD14(+) cells was analyzed by flow cytometry, the immunoregulatory function of DC was tested by mixed lymphocyte reaction (MLR). The change of IL-6 and VEGF as well as EPK and p38 signal pathway were analyzed by neutral antibody experiment and Western blot. The results showed that HUVEC isolated from human umbilical cord were characterized by spindle-shaped morphology, homogenous immunophenotypes (vWF(+)CD31(+)CD73(+)CD45(-)HLA-DR(-)CD86(-)CD34(low)), Dil-Ac-LDL incorporation ability and forming capillary-like structures. Following stimulation with granulocyte-macrophage colony-stimulating factor (GM-CSF) plus interleukin-4 (IL-4), HUVEC cocultures could inhibit the initial differentiation of CD14(+) monocyte to DC. Interestingly, IL-6 and VEGF enhanced the suppression effect of HUVEC on generation of DC via activation of the ERK or p38 mitogen activated protein kinase pathway. It is concluded that HUVEC are involved in DC development and can suppress the differentiation of monocyte to DC.
Cell Differentiation ; Coculture Techniques ; Dendritic Cells ; cytology ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Monocytes ; cytology

This study was purposed to investigate the regulatory effects of differentiating mesenchymal stem cells (MSC) on osteoclast formation. The MSC from mouse compact bones were cultured and induced into osteoblasts and adipocytes for one week. To test their regulatory effect on osteoclastogenesis, osteogenically differentiated and adipogenically differentiated MSC were co-cultured with CD11b(+) monocytes and osteoclasts were identified with in situ tartrate-resistant acid phosphatase (TRAP) staining. The results showed that differentiated MSC supported osteoclastogenesis but the osteoclast supporting capacity of osteogenically differentiated MSC decreased as compared with undifferentiated MSC. More interestingly, the adipogenically differentiated MSC significantly promoted osteoclasts formation when co-cultured with monocytes. It is concluded that the regulatory effect of MSC on osteoclast formation has changed while they have differentiated into different types of cells. The findings indicate that MSC may exert alternative effect on osteoclastogenesis by differentiation to descendant cells.
Adipogenesis ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Monocytes ; cytology ; Osteoblasts ; cytology ; Osteoclasts ; cytology

Objective To study the relativity between imageology and pathology during lung cancer,and estimate whether the lung cancer is preinvasive lesions,which can support evidences for the operation methods.Methods Clinical data of 624 patients who were diagnosed as lung adenocarcinoma and had solitary pulmonary nodule(diameter≤3 cm) were collected,all of them were scanned by thin layer CT scan(1 mm).The correlation between imageology and pathology data were analyzed.Results In 125 cases of GGO,the ratio of invasive lesions were 0 (0/72),6.1％ (3/49) and 100％ (4/4) in stage T1a,T1b and T1c respectively.In 285 cases of mGGO,if solid component was less than 0.5 cm,the ratio of invasive lesions were 1.7％ (1/58),6.9％ (2/29) and 50.0％ (2/4) in stage T~,T1b and Tic;but the ratio of invasive lesions were 81.3％ (13/16),94.1％ (96/102) and 97.4％ (74/76) respectively when the solid component was more than 0.5 cm.In 214 cases with solid nodules,the ratio of invasive lesions were 87.1％ (27/31),98.8％ (84/85) and 99.0％ (97/98) in stage T1 a,T1b and T1c.Conclusion The ratio of invasive lesions and solid component increased gradually along with the growing of tumor diameter in stage T1 lung cancer.CT imaging was highly correlated with the pathology diagnosis of preinvasive lesions and invasive lesions,which can be used as the guidance for operation methods.

OBJECTIVETo study the clinical features and to identify homeobox D13 (HOXD13) gene mutation of the affected individuals in a Chinese synpolydactyly (SPD) kindred.

METHODSClinical data and peripheral blood samples of SPD family members were obtained through field investigation. For every member of this pedigreeìthe fragment containing mutational hot spots of HOXD13 was amplified by PCR for mutation screening. To examine whether there is any other mutation within coding sequence of HOXD13, exon 1 and exon 2 of HOXD13 were also amplified by PCR. All the amplified fragments were electrophoresed on 2% agarose gels and then the mutant fragments were electrophoresed on 5% polyacrylamide gels to be separated. Purified PCR products of normal and selected mutant alleles were directly sequenced.

RESULTSComparing the HOXD13 coding sequence of the affected individuals with HOXD13 sequence in the GenBank and with that of the unaffected, an inserted segment coding 8 alanine residues within HOXD13 was found segregating with the disorder. This mutation is also termed polyalanine expansion. The 8-alanine expansion can be interpreted as a reduplication of normal alanines 5-12.

CONCLUSIONThe results suggest that synpolydactyly in this kindred may be caused by polyalanine expansion in HOXD13.

Base Sequence ; China ; DNA Mutational Analysis ; Female ; Homeodomain Proteins ; genetics ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Syndactyly ; genetics ; Transcription Factors ; genetics

OBJECTIVETo study the effect of Platycarya strobilacea Sieb. et Zucc (PSZ) extract on methuosis of human nasopharyngeal carcinoma CNE1 and CNE2 cells and explore the underlying mechanism.

METHODSCNE1 and CNE2 cells were treated with 1 mg/mL PSZ extract and the expressions of Rac1 mRNA and Rac1 protein were detected using RT-qPCR and Western blotting, respectively. Results CNE1 and CNE2 cells showed obvious morphological changes typical of methuosis following treatment with PSZ extract characterized by cell merging, accumulation of large cytoplasmic vacuoles, and membrane rupture without obvious changes in the nuclei. PSZ treatment resulted in up-regulated Rac1 mRNA and Rac1 protein expressions in the cells. Application of EHT 1864 obviously blocked the effect of PSZ extract in inducing methuosis in CNE1 and CNE2 cells.

CONCLUSIONPSZ extract can induce methuosis in CNE1 and CNE2 cells by inducing the overexpression of Rac1.