Objective To discuss the related factors of early pathological fracture after curettage of benign bone tumors in femoral shaft.Methods The clinical data of 47 patients with benign bone tumors in femoral shaft,treated by curettage with bone graft via the vastus lateralis approach from March 2004 to March 2011,were retrospectively analyzed.Thirteen patients of them presented with early pathological fracture after the curettage.In fracture group,there were 13 cases,11 males and 2 females,and the time from finishing curettage to fracture occurring ranged from 21 to 36 days.In non-fracture group,there were 34 cases,23 males and 11 females.The following data of fracture group and non-fracture group were compared and analyzed,such as specific value of absolute width of tumor and transverse diameter of bone shaft,specific value of defect width of bone window and sagittal diameter of bone shaft,defect length-width ratio of bone window,defect morphology of bone window,classification of bone tumor,violence of causing injury and compliance to medical advice.Results The average defect length-width ratio of bone window in fracture group was 3.72±3.58,in non-fracture group was 2.67±6.35.For classification of tumor,in fracture group 1 case was in incubation period,6 in active period,6 in invasion period; in non fracture group 21 cases were in incubation period,10 in active period,and 3 in invasion period.Four cases in fracture group had poor compliance to medical advice,and 9 in non-fracture group had good compliance.Between two groups,there were no statistical differences in specific value of absolute width of tumor and transverse diameter of bone shaft,specific value of defect width of bone window and sagittal diameter of bone shaft,and defect morphology of bone window.Conclusion When defect length-width ratio of bone window is larger than 4,the classification of tumor causes expanded incisal edge,and the cortical bone was damaged extensively,there are more possibilities for pathological fracture.

Objective To investigate the effect of different concentrations of human adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.Methods 0.3 ml fat tissue,derived and refined from clinical liposuction patients.was mixed with different concentrations of SVFs as 5 × 105/ml in Group A,or 1 × 106/ml in Group B,or 2 × 106/ml in Group C,or completely medium in control group D.Then the mixture was injected randomly under the back skin of 6 nude mice.The transplanted fat tissue in four groups was harvested at 3 months after implantation.Wet weight of fat grafts was measured for macroscopic aspects.After HE staining,blood vessel density,viable adipocytes and fibrous proliferation were counted respectively for histological evaluation.Results The wet weight of fat grafts in group B(81.670 ± 7.528) mg was significantly higher than that in group A、C、D [ (60.000 ±6.325 ) mmg、(68.330 ± 7.528 ) mg、(48.330 ± 7.528 ) mg,respectively,P ＜ 0.05 ) ],but the difference between group A and group C was not statistically significant(P ＞ 0.05).The grafts in group A 、B and C had significantly higher blood vessel density than those in the control group D,whereas blood vessel density was the highest in group B( P ＜ 0.05 )and there was no significant difference between group A and C (P ＞0.05).Compared with group A,C and D,histological analysis revealed that the fat grafts in group B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis (P < 0.05 ).However,fibrosis counts were significant lower in group A,B and C than those in group D( P <0.05),and there was no significant difference between group A and C(P ＞0.05).Conclusions The human isolated SVFs has the advantages to improve the survival rate of fat transplantation,and the magnitude of 1 × 106/ml is more practical and safe,indicating a wide clinical application in the future.

Objective To investigate the effect of different concentrations of human adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.Methods 0.3 ml fat tissue,derived and refined from clinical liposuction patients.was mixed with different concentrations of SVFs as 5 × 105/ml in Group A,or 1 × 106/ml in Group B,or 2 × 106/ml in Group C,or completely medium in control group D.Then the mixture was injected randomly under the back skin of 6 nude mice.The transplanted fat tissue in four groups was harvested at 3 months after implantation.Wet weight of fat grafts was measured for macroscopic aspects.After HE staining,blood vessel density,viable adipocytes and fibrous proliferation were counted respectively for histological evaluation.Results The wet weight of fat grafts in group B(81.670 ± 7.528) mg was significantly higher than that in group A、C、D [ (60.000 ±6.325 ) mmg、(68.330 ± 7.528 ) mg、(48.330 ± 7.528 ) mg,respectively,P ＜ 0.05 ) ],but the difference between group A and group C was not statistically significant(P ＞ 0.05).The grafts in group A 、B and C had significantly higher blood vessel density than those in the control group D,whereas blood vessel density was the highest in group B( P ＜ 0.05 )and there was no significant difference between group A and C (P ＞0.05).Compared with group A,C and D,histological analysis revealed that the fat grafts in group B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis (P < 0.05 ).However,fibrosis counts were significant lower in group A,B and C than those in group D( P <0.05),and there was no significant difference between group A and C(P ＞0.05).Conclusions The human isolated SVFs has the advantages to improve the survival rate of fat transplantation,and the magnitude of 1 × 106/ml is more practical and safe,indicating a wide clinical application in the future.

OBJECTIVETo explore the possibility of building tissue-engineered adipose tissue and looking for a new approach for the repair of soft tissue defects.

METHODSThe cells using enzymatic digestion from human liposuction part of the lipid extract were used as adipose tissue-derived cells and labeled with DiI fluorescent marker, the induced group using I collagen scaffold material as a carrier, the induced cell were planted into left back subcutaneously in nude mice at 1 x 10(7)/ml cell density, in the uninduced group cells were not induced by any, in the same cell density and type I collagen scaffold composite inoculated in nude right mouse back skin, the blank control group I collagen scaffold gaps in nude mice inoculated subcutaneously center of the neck, each of the six mice; Remove implants after 12 weeks and judge the adipogenic capacity through general and fluorescence microscopy, wet - determination, histological detection and oil red O staining qualitative.

RESULTSThe primary source of fat cultured stem cells, similar to the fibroblast morphology, and has a strong proliferative capacity. In the role of adipose differentiation medium, it can be the mature fat cells in which cytoplasmic lipid droplets gather, oil red O staining was positive. In the induced group, newborn tissue were found in the experimental groups of nude mice and its average weight is about 0.020 g. Conventional pathological slices and oil red O staining confirmed it is mature adipose tissue, the fluorescence staining positive confirm them are exogenous. Uninduced group newborn tissue are found in the experimental groups of nude mice and its average weight is about 0.014 g. Conventional pathological slices and oil red O staining confirmed it include some mature adipose tissue, the fluorescence staining positive confirm them are exogenous. Two groups of the new wet weight with have statistical significance (P < 0.01); gaps in the control group no new organization formed.

CONCLUSIONSThe cells using enzymatic digestion from human liposuction part of the lipid extract are adipose tissue-derived cells. The cells can be as seed cells and with solid scaffold of collagen type I it can become fat tissue in vivo successfully.

Adipose Tissue ; cytology ; metabolism ; Animals ; Cell Culture Techniques ; Collagen Type I ; biosynthesis ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Stem Cells ; cytology ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo study the cellular compatibility of type I collagen scaffold and human adipose-derived stem cells (ADSC(S)) in order to explore appropriate scaffold materials for adipose tissue engineering.

METHODSThe morphology and function of the ADSC(S) were observed by inverted phase contrast microscope, scanning electron microscope and XTT assay when cocultured type I collagen scaffold with ADSC(S) in vitro. Cells adhesive rates were also calculated.

RESULTADSC(S) were able to attach, grow and proliferate well on the scaffolds.

CONCLUSIONcollagen I scaffold exhibits excellent cellular compatibility and can be used as a vehicle for adipose tissue engineering.

Adipocytes ; cytology ; Adult Stem Cells ; cytology ; Biocompatible Materials ; chemistry ; Cells, Cultured ; Coculture Techniques ; Collagen Type I ; chemistry ; Humans ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo investigate the dedifferentiation of mature adipocytes and the possibility of adipose tissue engineering using dedifferentiated adipocytes.

METHODSHuman adipose tissue was harvested from healthy women undergoing abdominal liposuction procedures, and mature adipocytes were isolated with enzymatic digestion and cultured by ceiling adherent culture method, using the third-passage cells for subsequent experiment. The cells were cultured in adipogenic, chondrogenic or osteogenic media, and Oil red-O staining, Alcian blue staining and Alizarin red staining were used for dedifferentiation identification. The third-passage cells labeled with DiI were mixed with fibrin glue and injected subcutaneously on one side of the nude mouse back (n=6), and fibrin glue solution without cells as control was injected on the other side (n=6). After 8 weeks of cell implantation, the specimens were harvested for general observation and histological, Oil red-O staining and fluorescent microscope analyses.

RESULTSMature adipocytes were round, unilocular cells. After ceiling adherent culture, the adipocytes underwent morphological changes into fibroblast-like cells indicating their dedifferentiation. The dedifferentiated adipocytes were induced for adipogenic, chondrogenic and osteogenic differentiation in specified media. Eight weeks after the cell injection in nude mice, newly formed tissue occurred which was identified as mature adipose tissue. The implants without cells were completely absorbed in the control group.

CONCLUSIONMature adipocytes can dedifferentiate in vitro culture, and the dedifferentiated adipocytes are capable of differentiating into adipogenic, chondrogenic and osteogenic lineages. Adipose tissue engineering can be achieved in vivo using the dedifferentiated adipocytes as the seed cells.

Adipocytes ; cytology ; Adipogenesis ; Adult ; Animals ; Cell Dedifferentiation ; Chondrogenesis ; Female ; Humans ; Mice ; Mice, Nude ; Microscopy, Fluorescence ; Osteogenesis ; Time Factors ; Tissue Engineering ; methods

OBJECTIVETo establish a real-time reverse transcription-polymerase chain reaction (RT-PCR) for quantitative detection of E2A-PBX1 fusion gene mRNA in acute lymphoblastic leukemia (ALL) children and to explore its clinical significance in minimal residual disease monitoring and prognosis evaluation.

METHODSReal-time RT-PCR was used to quantitatively detect the mRNA expression of E2A-PBX1 gene in 11 newly diagnosed ALL patients at diagnosis (11 cases), complete remission (11 cases) and periods of relapse (3 cases). Ten children with normal bone marrow cell morphology and without hematopathy or tumor diseases were used as the control group.

RESULTSThe median expression levels of E2A-PBX1 fusion gene in the ALL group at diagnosis and the relapse group were significantly higher than in the control and complete remission groups (P<0.01). Compared with E2A-PBX1 negative patients on day 33 during induction of remission, the recurrence rate increased and disease free survival rate at 3 year decreased significantly in E2A-PBX1 positive patients decreased (P<0.05).

CONCLUSIONSMeasurement of E2A-PBX1 levels by real-time RT-PCR is useful for monitoing minimal residual disease, prediction of relapse and individual treatment. The expression level of E2A-PBX1 gene on day 33 during induction of remission can be used for prognosis evaluation.

Adolescent ; Child ; Child, Preschool ; Female ; Homeodomain Proteins ; genetics ; Humans ; Male ; Neoplasm, Residual ; diagnosis ; Oncogene Proteins, Fusion ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; Prognosis ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVETo explore the dedifferentiation phenomenon of human mature adipocytes cultured in vitro and to discuss the possibility of using dedifferentiation adipocytes (DA) as seed cells.

METHODSMature adipocytes and ASCs were harvested from human fat aspirates. Mature adipocytes were cultured and induced to DA by ceiling adherent culture method. Cell morphology were observed during the whole process. Viabilities of DA and ASCs were compared by MTT chromatometry and cell growth curves were drawn based on it. Cell surface markers of DA and ASCs were detected by flow cytometry. The adipogenic, osteogenic and chondrogenic ability of DA and ASCs were assessed by oil red O staining, alizarin bordeaux staining and alcian blue staining, respectively.

RESULTSHuman mature adipocytes can dedifferentiate into fibroblast-shaped DA. MTT chromatometry assay demonstrated that DA and ASCs both had strong reproductive activity, with no significant difference between them. Flow cytometry assay demonstrated that both DA and ASCs expressed HLA-ABC, CD29 and CD44, while didn't express CD45, CD34 and CD106. After two weeks of adipogenic differentiation, lipid droplets could be displayed by oil red O staining in both DA and ASCs. After two weeks of osteogenic differentiation, calcium salts mineralization in DA and ASCs could be detected by alizarin bordeaux staining. After two weeks of chondrogenic differentiation, matrix of cartilage cells in DA and ASCs could be detected by alcian blue staining.

CONCLUSIONSMature adipocytes can be dedifferentiated into DA in vitro. DA has strong reproductive activity, as well as osteogenic, chondrogenic ability and strong adipogenic ability. It expresses some of the stem cell-related cell surface proteins and is a promising seed cell for adipose tissue engineering.

Adipocytes ; cytology ; Adipose Tissue ; cytology ; Adolescent ; Adult ; Cell Culture Techniques ; Cell Dedifferentiation ; Cells, Cultured ; Female ; Humans ; Male ; Stem Cells ; cytology ; Young Adult

OBJECTIVETo compare the adipogenic differentiation capacity of dedifferentiated adipocytes cells (DA) and adipose-derived stem cells (ASCs) in vivo, so as to select good adipogenic seed cells for tissue engineering.

METHODSMature adipocytes and ASCs were isolated by means of enzymatic digestion from the liposuction aspirate. Then the DA cells were acquired by ceiling adherent culture of mature adipocytes and the 3rd passage cells were used. The DA cells and ASCs were cultured with fibrin glue in vitro respectively. The compatibility of scaffold with cells was detected by microscopy and scanning electron microscopy. The scaffold-cell composite was also labeled by DiI. The composite was injected subcutaneously on the nude mice back, respectively (DA-FG group, n = 8; ASCs-FG group, n = 8; sham FG group, n = 8). 8 weeks after implantation, the newly formed tissue was taken out for general observation and histologic study.

RESULTSMature adipocytes were transferred to DA cells with spindle shape, like fibroblast. The ASCs were also spindle. Three days after culture of cell-scaffold composite in vitro, the cells grew well. 8 weeks after implantation, the newly formed tissue was found under the skin both in DA-FG and ASCs-FG groups, but not in sham FG group. The newly formed tissue was mature fat tissue and originated from the seed cells. The average wet weight of the new-formed tissue was higher in DA-FG group than that in ASCs-FG group. The average fibrosis ratio was lower in DA-FG than in ASCs-FG group.

CONCLUSIONSThe tissue-engineered adipose tissue can be achieved with DA cells and ASCs as seed cells. Compared with ASCs, the new-formed fat tissue with DA has a higher wet weight and lower fibrosis ratio.

Adipocytes ; cytology ; Adipose Tissue ; cytology ; Adult ; Animals ; Cell Differentiation ; Cells, Cultured ; Female ; Humans ; Mice ; Mice, Nude ; Stem Cells ; cytology ; Tissue Engineering

OBJECTIVETo investigate the effect of different concentrations of human adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.

METHODS0.3 ml fat tissue, derived and refined from clinical liposuction patients, was mixed with different concentrations of SVFs as 5 x 10(5)/ml in Group A, or 1 x 10(6)/ml in Group B, or 2 x 10(6)/ml in Group C, or completely medium in control group D. Then the mixture was injected randomly under the back skin of 6 nude mice. The transplanted fat tissue in four groups was harvested at 3 months after implantation. Wet weight of fat grafts was measured for macroscopic aspects. After HE staining, blood vessel density, viable adipocytes and fibrous proliferation were counted respectively for histological evaluation.

RESULTSThe wet weight of fat grafts in group B (81.670 +/- 7.528) mg was significantly higher than that in group A, C, D [(60.000 +/- 6.325) mg, (68.330 +/- 7.528) mg, (48.330 +/- 7.528) mg, respectively, P < 0.05)], but the difference between group A and group C was not statistically significant (P > 0.05). The grafts in group A, B and C had significantly higher blood vessel density than those in the control group D, whereas blood vessel density was the highest in group B (P < 0.05) and there was no significant difference between group A and C (P > 0.05). Compared with group A, C and D, histological analysis revealed that the fat grafts in group B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis (P < 0.05). However, fibrosis counts were significant lower in group A, B and C than those in group D (P < 0.05), and there was no significant difference between group A and C (P > 0.05).

CONCLUSIONSThe human isolated SVFs has the advantages to improve the survival rate of fat transplantation, and the magnitude of 1 x 10(6)/ml is more practical and safe, indicating a wide clinical application in the future.

Adipose Tissue ; cytology ; transplantation ; Adult ; Animals ; Cells, Cultured ; Female ; Graft Survival ; Humans ; Male ; Mice ; Mice, Nude ; Stromal Cells ; transplantation