The osteogenic in vitro effect of low intensity pulsed ultrasound (LIPUS) on SD rat adipose-derived stem cells (ADSCs) was investigated. Rat ADSCs underwent LIPUS (intensity=100 mW/cm(2)) or sham exposure for 8 min per treatment once everyday in vitro, and then the alkaline phosphatase (ALP) activity and mineralized nodule formation were assessed to evaluate the osteogenic effect of LIPUS on ADSCs. To further explore the underlying mechanism, the osteogenic-related gene mRNA expression was determined by using reverse transcriptase-polymerase chain reaction (RT-PCR) at 1st, 3rd, 5th, 7th day after exposure repectively. Westen blot was used to evaluate the protein expression levels of two osteogenic differentiation associated genes at 7th and 14th day repectively. It was found that ALP activity was increased after LIPUS exposure and LIPUS resulted in mineralized nodule formation of ADSCs in vitro. LIPUS-treated ADSCs displayed higher mRNA expression levels of runt-related transcription factor 2 (Runx2), osteocalcin (OCN), ALP and bone sialoprotein (BSP) genes than controls, and the protein levels of Runx2 and BSP were also increased. The results suggested that LIPUS may induce the osteogenic differentiation of ADSCs in vitro.

Objective To analyze the consistency of the SYSMEX UF1000i automatic urinary sediment analyzer,Arkray AX-4030 urine dry chemistry analyzer and optical microscope in detecting urine erythrocyte.Methods The fresh urine specimens from 427 patients were randomly extracted and tested by the SYSMEX UF1000i automatic urinary sediment analyzer,urine dry chemistry analyzer and OLUMPUS Arkray AX-4030 optical microscope.Then the consistency of the results for detecting urine erythrocyte was compared among three kinds of detection method.Results With the microscopic examination as control,the sensitivity and spe-cificity of the SYSMEX UF1000i automatic urinary sediment analyzer for detecting urine erythrocyte were 82.84% and 86.35% re-spectively,which of the Arkray AX-4030 urine dry chemistry analyzer were 89.55% and 83.96% respectively.There was a high consistency between the SYSMEX UF1000i automatic urinary sediment analyzer and the optical microscope for detecting urine e-rythrocyte and the Kappa value was 0.580.There was also a high consistency between the Arkray AX-4030 urine dry chemistry analyzer and the optical microscope for detecting urine erythrocyte and the Kappa value was 0.625,while the consistency between the SYSMEX UF1000i automatic urinary sediment analyzer and the Arkray AX-4030 urine dry chemistry analyzer was weaker and the Kappa value was 0.324.Conclusion With the detection by the SYSMEX UF1000i automatic urinary sediment analyzer and the Arkray AX-4030 urine dry chemistry analyzer as a screening test,it should need to combine with the optical microscopy to conduct recheck for providing the effective and reliable test results quickly and accurately.

Some studies indicate that adipose derived stem cells (ADSCs) can differentiate into adipogenic, chondrogenic, myogenic, and osteogenic cells in vitro. However, whether ADSCs can be induced to differentiate into neural cells in vitro has not been clearly demonstrated. In this study, the ADSCs isolated from the murine adipose tissue were cultured and transfected with the EGFP gene, and then the cells were induced for neural differentiation. The morphology of those ADSCs began to change within two days which developed into characteristics of round cell bodies with several branching extensions, concomitantly expressing EGFP fluorescence. Approximately 60% of the total cell populations were bipolar or multipolar in shape. Some of them appeared to make contact with their neighboring cells. RT-PCR, Western blot and Immunocytochemistry revealed that the expression levels of the markers of neurons and oligodendrocytes such as MAP2, NF-70, Neu N and RIP upon neural induction were increased, but the expression of the special marker of astrocytes, GFAP, was undetectable until 96 h after induction when a small signal was observed. It was concluded that the ADSCs transfected with EGFP possessed the ability to undergo morphologic and phenotypic changes consistent with neural differentiation in vitro. It suggests that these cells might provide an ideal source for further stem cell research with possible therapeutic application for spinal cord injury.

Some studies indicate that adipose derived stem cells(ADSCs)can differentiate into adipogenic,chondrogenic,myogenic,and osteogenic cells in vitro.However,whether ADSCs can be induced to differentiate into neural cells in vitro has not been clearly demonstrated.In this study,the ADSCs isolated from the murine adipose tissue were cultured and transfected with the EGFP gene,and then the cells were induced for neural differentiation.The morphology of those ADSCs began to change within two days which developed into characteristics of round cell bodies with several branching extensions,concomitantly expressing EGFP fluorescence.Approximately 60% of the total cell populations were bipolar or multipolar in shape.Some of them appeared to make contact with their neighboring cells.RT-PCR,Western blot and Immanocytochemistry revealed that the expression levels of the markers of neurons and oligodendrocytes such as MAP2,NF-70,Neu N and RIP upon neural induction were increased,but the expression of the special marker of astrocytes,GFAP,was undetectable until 96 h after induction when a small signal was observed.It was concluded that the ADSCs transfected with EGFP possessed the ability to undergo morphologic and phenotypic changes consistent with neural differentiation in vitro.It suggests that these cells might provide an ideal source for further stem cell research with possible therapeutic application for spinal cord injury.

The osteogenic in vitro effect of low intensity pulsed ultrasound (LIPUS) on SD rat adipose-derived stem cells (ADSCs) was investigated.Rat ADSCs underwent LIPUS (intensity=100 mW/cm2) or sham exposure for 8 min per treatment once everyday in vitro,and then the alkaline phosphatase (ALP) activity and mineralized nodule formation were assessed to evaluate the osteogenic effect of LIPUS on ADSCs.To further explore the underlying mechanism,the osteogenic-related gene mRNA expression was determined by using reverse transcriptase-polymerase chain reaction (RT-PCR) at 1st,3rd,5th,7th day after exposure repectively.Westen blot was used to evaluate the protein expression levels of two osteogenic differentiation associated genes at 7th and 14th day repectively.It was found that ALP activity was increased after LIPUS exposure and LIPUS resulted in mineralized nodule formation of ADSCs in vitro.LIPUS-treated ADSCs displayed higher mRNA expression levels of runt-related transcription factor 2 (Runx2),osteocalcin (OCN),ALP and bone sialoprotein (BSP) genes than controls,and the protein levels of Runx2 and BSP were also increased.The results suggested that LIPUS may induce the osteogenic differentiation of ADSCs in vitro.

CD146 Antigen ; immunology ; Fibroblasts ; immunology ; pathology ; Humans ; Scleroderma, Systemic ; drug therapy ; immunology ; pathology