3.Up-expression of peroxisome proliferator-activated receptor-γand of retinoid X receptor-αenhances the inhibitory effect on leukemia cells
Yun GUO ; Ruicheng XU ; Tao HE
Journal of Leukemia & Lymphoma 2009;18(3):131-133
Objective To investigate the relationship between the expression of peroxisome proliferators-activated recepter-γ (PPARγ) and retinoid X receptor-or (RXRα) and the inhibitory effect of PLAB, ligand of PPARγ and 9-cisRA, ligand of RXRα on growth of human leukemia cell lines (HL-60, K562and U937) in vitro. Methods The antiproliferative effect was evaluated by MTT assay. The mRNA expression of PPARγ and RXRα was semi-quantified by RT-PCR. Results PPARγ and RXRα mRNA was both expressed in HL-60, K562 and U937 cells, and the expression in HI,-60 was significantly higher than that in K562 and U937. The significant inhibitory effect on the growth of HL-60 cells was observed in K562 and U937 cells. The combination group showed more inhibitory effect in HL-60 cells than PLAB alone(P<0.05).PLAB significantly up-regulates the expression of PPARγ in HL-60 cells, the expression of PPARγ and RXRα were higher in combination group than PLAB alone (P<0.05). Conclusion The expression of PPARγand RXRα in HL-60, K562 and U937 cell lines predicts their response to PLAB and 9-cisRA treatment, and the inhibitory effect is different in these three kinds of cell lines, which may be related to their ligandsmediated signal pathway.
4.The effects of high-intensity pulsed electromagnetic field on proliferation and differentiation of neural stem cells of neonatal rats in vitro.
Depeng, MENG ; Tao, XU ; Fengjin, GUO ; Weifeng, YIN ; Tao, PENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(6):732-6
The effects of high-intensity pulsed electromagnetic stimulation (HIPEMS) on proliferation and differentiation of neonatal rat neural stem cells in vitro were investigated. Neural stem cells derived from neonatal rats were exposed to 0.1 Hz, 0.5-10 Tesla (T) [8 groups of B-I, respectively], 5 stimuli of HIPEMF. The sham exposure controls were correspondingly established. Inverted phase contrast microscope was used to observe the cultured cells, MTT assay to detect the viability of the cells as expressed by absorbance (A) value, and flow cytometry to measure differentiation of neural stem cells. The results showed that A values of neural stem cells in both 3.0 T and 4.0 T groups were significantly higher than the other groups 24 to 168 h post HPEMS, indicating a strong promotion of the growth of neural stem cells (P<0.05). The A values of neural stem cells in the 6.0 T, 8.0 T, and 10.0 T groups were lower than the sham exposure control group, indicating a restraint of the growth of neural stem cells. The rate of neuron-specific enolase-positive neurons revealed by flow cytometry in HPEMS groups was the same as that in control group (P>0.05). It was suggested that 0.1 Hz, 5 pulses stimulation of HPEMS within certain scale of intensity (0.5-10.0 T), significantly promoted the growth of neural stem cells with the rational intensity being 4.0 T.
5.Effects of magnetic stimulation on proliferation and differentiation of endogenous neural stem cells/progenitor cells after spinal cord injury in rats
Tao XU ; Fengjing GUO ; Anmin CHEN ; Xiaolin HUANG ; Tiecheng GUO
Chinese Journal of Physical Medicine and Rehabilitation 2010;32(1):2-5
Objective To investigate the effects of magnetic stimulation (MS) on the proliferation and differentiation of endogenous neural stem cells (NSCs)/progenitor cells after spinal cord injury (SCI) in rats. Methods Forty-six Wistar rats were used, of which 40 were used to make an animal model of spinal cord injury (SCI) by administering a 10 g x 12.5 cm impact at the T8 level. The other 6 served as the normal controls. The SCI model rats were evenly divided into a magnetic stimulation (MS) group ( n = 20) and a control group ( n = 20). The rats in the MS group received 0.5 Hz and 1.44 T magnetic stimulation 24 h post injury, then 30 pulses per day for 7 days. The rats in the other groups were not exposed to MS. The scale of Basso, Beatti and Bresnahan (BBB) was used to assess hindlimb neurological function. Rats were sacrificed at the 24th hour, and at the 1st, 4th and 8th weeks after SCI. The ratio of nestin to microtubule associated protein 2 (MAP2)/nestin in the cells of the spinal cord was determined by immunofluorescence. Results The BBB scores in the MS group were signifi-cantly higher than those of the control group at 1, 4 and 8 weeks post SCI. Nestin and the MAP2/nestin ratios were mild in the normal spinal cords, but increased after SCI. They were higher in the MS group than that in the control groups at all time points. Conclusions MS can promote nestin expression in the spinal cord after SCI and facili-tate neural differentiation.
6.Comparison of routine test kits of creatinine in human serum with reversed-phase high-pressure liquid chromatographic assay
Jing XU ; Qing-Tao WANG ; Guo-Bin XU ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
98.95%.The results of certified reference materials were consistent with the target value,and average deviation was -0.31%~1.35%.HPLC was served as the independent variable.When ereatinine was 200 ?mol/L,Bias of Beckman LX20 system,Vitros dry chemistry system and enzymatic method were -10%~13%,-13%~14% and -20%~10%,respectively.Bias of enzymatic method results was mostly negative,when creatinine was
7.The effects of low intensity pulsed ultrasound on the proliferation and osteogenic differentiation of rat adipose tissue-derived stem cells
Ting JIANG ; Fengjing GUO ; Tao XU ; Keqing YANG
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(6):408-412
Objective To investigate the biological effects of low intensity pulsed ultrasound (LIPUS) on the proliferation and osteogenic differentiation of adipose tissue-derived stem cells (ADSCs) in vitro.Methods Primary ADSCs were harvested from the inguinal fat pads of 4-week-old female Sprague-Dawley rats,cultured in vitro and purified by magnetic-activated cell sorting.Surface ADSC markers were identified by flow cytometry.LIPUS at 100 mW/cm2 was used to stimulate the cultured cells.Flow cytometry was performed for cell cycle analysis.Cellular proliferation was evaluated via CCK8 chromatometry,and a proliferation index was calculated.ADSCs were assigned to 4 groups:a negative control group,a LIPUS group,an osteoinduction group and a LIPUS plus osteoinduction group,and treated accordingly.Alkaline phosphatase (ALP) activity was determined at the 7th and 14th day in each group,and calcium nodes were marked by Von Kossa staining.The levels of osteogenic differentiation in the different groups were evaluated.Results The ADSCs of passage 3 expressed CD 34low,and CD29high CD44high,which was consistent with the characteristics of ADSC surface markers.Proliferation was upregulated significantly in the LIPUS group compared with the negative control group.ALP activity was also elevated significantly and it resulted in mine-ralization.The highest mineralization rate was observed in the LIPUS plus osteoinduction group.Conclusions LIPUS not only can stimulate the proliferation of rat ADSCs,it also promotes their osteogenic differentiation.
8.Serum transforming growth factor beta 1 contributes to bone healing in patients with bone fractures combined with spinal cord injuries
Sen YANG ; Hailong WANG ; Weibin SHENG ; Tao XU ; Hailong GUO
Chinese Journal of Tissue Engineering Research 2015;(2):165-169
28 days after injury (P < 0.05). In the fracture+spinal cord injury group, the level of serum transforming growth factor beta 1 had a rapid increase on the 7th day, and reached the peak on the 14th day, and then, this level had no significant decrease until the 28th day. In the simple fracture group, the level of serum transforming growth factor beta 1 began to increase on the 2nd day, reached the peak on the 7th day, and then decreased gradualy. Remarkable changes of serum transforming growth factor beta 1 levels in patients with bone fracture combined with spinal cord injury may be associated with fracture healing in different periods.
9.Establishment and application of human CHO/NTR1 system.
Guo ZHANG ; Tao SUN ; Huijuan LIU ; Guojun NIU ; Feng XU
Acta Pharmaceutica Sinica 2014;49(9):1273-8
Abstract: Neurotensin receptor-1 (NTR1), which can stimulate the intracellular cascade signal pathway, belongs to the large superfamily of G-protein coupled receptors. NTR1 is related to the occurrence and development of several kinds of diseases. In order to screen the inhibitors for the cancers associated with NTR1 protein, we established a CHO (Chinese hamster ovary) cell line in which human neurotensin receptor-1 was highly expressed. The method is to construct the recombinant plasmid which was lysed with the hNTR1 gene and transfect it into CHO cells. After selected with G418, the cell line was evaluated by Western blotting analysis and calcium flux assays. Through the calcium flux assays on FlexStation 3, we got the EC50 value of neurotensin peptide which is the natural NTR1 agonist, and the IC 50 value of SR48692 which is the known NTR1 antagonist. The established human CHO/NTR1 cell line can be used to study the profile of NTR1 biological activity and further screen of NTR1 antagonists and agonists.
10.Effect of hydrogen sulifde on the expression of CSE, NF-κB, and IL-8 mRNA in GES-1 cells withHelicobacter pylori infection
Canxia XU ; Yingchun WAN ; Tao GUO ; Xiong CHEN
Journal of Central South University(Medical Sciences) 2013;38(10):977-983
Objective: To investigate the effect of hydrogen sulifde (H2S) on the expression of CSE, NF-κB, and IL-8 mRNA in GES-1 cells withHelicobacter pylori (H. pylori) infection and to explore its mechanism on gastric mucosa inlfammation caused byH. pylori.
Methods: GES-1 cells were cultured for 24 h and divided into a control group (neitherH. pylori nor NaHS), anH. pylori group, a NaHS group (which was further divided into 4 groups at 50, 100, 200, or 400 μmol/L NaHS), andH. pylori + NaHS group (which was further divided into 4 groups at 50, 100, 200, or 400 μmol/L NaHS). Each group was then cultured for 3, 6, or 12 h. The expression of CSE, NF-κB, and IL-8 mRNA was measured by RT-PCR, and their correlation was analyzed. Results: The expression of CSE, NF-κB, and IL-8 mRNA in GES-1 cells in theH. pylori group was higher than that in the control group. The expression of CSE in the 200 μmol/L NaHS group and 400 μmol/L NaHS group was lower than that of the control group (P<0.05), whereas the expression of NF-κB and IL-8 in all NaHS groups had no statistical differences compared with the control group (P>0.05). The expression of CSE, NF-κB, and IL-8 mRNA in all groups of NaHS,H. pylori + 200 μmol/L NaHS group, andH. pylori + 400 μmol/L NaHS group was lower than that in theH. pylori group (P<0.05). There was positive correlation among the expressions of CSE, NF-κB, and IL-8 mRNA in theH. pylori group, theH. pylori + 200 μmol/L NaHS group, and theH. pylori +400 μmol/L NaHS group (P<0.05).
Conclusion:H. pylori can induce NF-κB and IL-8 mRNA expression and upregulate CSE mRNA expression. At 200 and 400 μmol/L, NaHS can suppressH. pylori-induced NF-κB and IL-8 mRNA expression and ameliorate the morphology ofH. pylori-induced GES-1 injury, which may protect gastric epithelial cells byH. pylori infection.