ObjectiveTo explore the effect of bFGF-chitosan carriers on inducing bone marrow-derived mesenchymal stem cells (MSCs) to differentiate into nerve cells.MethodsMSCs were detected by immunohistochemistry and Western blot after they were induced by bFGF-chitosan carriers to differentiate into neurons. The MTT chromometry assay was carried out to determine cell viability.ResultsThe proportion of express neural stem cells marker Nestin, and neuronal markers class Ⅲ β-tubulin and MAP-2 was 83.54% after MSCs induced by bFGF-chitosan carriers.ConclusionbFGF-chitosan carriers can induce MSCs to differentiate into nerve cells with a high percentage.

BACKGROUND:Neural stem cells (NSCs) hold self-renewal and multi-directional differentiation potential. NSCs differentiation into neurons in high proportion under induction conditions exhibits broad application prospect. OBJECTIVE:To explore the effect of basic fibroblast growth factor (bFGF)-chitosan carrier on the NSCs differentiation into neurons in vitro, and whether the differentiated neurons could form synaptic-like connection with myocytes. METHODS:After purification, the NSCs were co-cultured with chitosan, soluble bFGF or bFGF-chitosan carrier. After 7-day induction, the NSCs differentiation into neurons was observed by immunofluorescence staining of beta tubulin Ⅲ. The NSCs differentiation into cholinergic neurons was observed through double immunofluorescence staining of ChaT and beta tubulin Ⅲ. The synaptic-like connection between the neurons and myocytes was observed by triple staining of beta tubulin Ⅲ and MHC. RESULTS AND CONCLUSION:The percentage of differentiated neurons in the bFGF-chitosan carrier group was 74%, which was significantly higher than that in the other two groups. Additionally, the synaptic-like connection formed between the differentiated neurons and myocytes. To conclude, the bFGF-chitosan carrier promotes the NSCs differentiation into neurons to form synaptic-like connection with the co-cultured myocytes.

Objective To silence the expression of CTGF by small interfering RNA technology,to observe the influence on fibroblast?like synovial cell apoptosis and several apoptosis?related genes,and to explore the mechanism of action of CTGF in rheumatoid arthritis synovial lesions. Methods Effective CTGF siRNA was screened through real?time PCR. The influence of CTGF siRNA on FLS apoptosis was detected with FITC?PI double staining by flow cytometry. bax,bcl?xl and survivin were detected using real?time PCR when CTGF mRNA has been silenced. Results Compared with other 2 groups of oligo and NC oligo,H1 oligo exhibited the strongest interfering action to CTGF(inhibition ratio>70%),so that it is selected as the effective target gene sequence for the following experiment. Apoptosis of FLS induced by serum deprivation was significantly decreased in the presence of exogenous CTGF. When expression of the CTGFgene was knocked down in FLS,FLS apoptosis was significantly increased,and expres?sion levels of survivin mRNA were decreased significantly(P<0.01). Conclusion FLS survival is positively regulated by CTGF,which may through the sustaining the expression of survivin.

Objective To explore the effect of basic fibroblast growth factor (bFGF)-chitosan carriers on neural differentiation of neural stem cells (NSCs). Methods NSCs were isolated from spinal cord of a neonatal Wistar rat and cultured. Purity of cultured NSCs was identified with Nestin immunofluorescent staining. The 10 mg/ml chitosan carriers, 20 ng/ml bFGF or 10 mg/ml bFGF-chitosan carriers were added into medium of P3~P4 NSCs respectively. NSCs were observed with immunofluorescent staining: 3 days after incubation with Nestin and β-tubulin III; 7 days after incubation with microtubule-associated protein-2 (MAP2), glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP); and 14 days after incubation with synapsin-1 and MAP2. The electrophysiological activity of cells was detected with MED64. Results 3 days after incubation, all the NSCs differentiated into Nestin+/β-tubulin III+, and the length of neurofilament was the highest in those co-cultured with bFGF-chitosan carriers. 7 days after incubation, NSCs differentiated into MAP2+, GFAP+ and MBP+, and more NSCs differentiated into MAP2+ with bFGF-chitosan carriers. 14 days after incubation, NSCs differentiated with bFGF-chitosan carriers express synapsin-1+/MAP2+ and showed electrophysiological activity. Conclusion bFGF-chitosan carriers can induce NSCs to differentiate into neuron with high percentage and the differentiated neurons can form synapses with electrophysiology activity.

Objective To explore the induction of basic fibroblast growth factor (bFGF)-chitosan carrier transforming the adult rat bone marrow mesenchymal stem cells (rMSCs) into nerve cells. Methods rMSCs were detected qualitatively and counted quantitatively by immunohistochemistry after they were induced into nerve cells, such as neural stem cells neurons and astrocytes. The methyl thiazolyl tetrazolium (MTT) chromometry assay was carried out to determine the cell viability. Results rMSCs induced by bFGF-chitosan carrier expressed neural stem cell marker nestin, neuron marker β-tubulin Ⅲ and astrocytes marker glial fibrillary acidic protein (GFAP). Nestin expressed more in the bFGF-chitosan group, and reached its maximum (49.40%) at the 9th day. Conclusion bFGF-chitosan carrier can induce the adult rMSCs differentiate into neural stem cells in a high proportion.

Objective To estimate the value of BIOMED-2 primers for the detection of T cell receptor γ (TCR-γ) gene rearrangements in different types of specimens from patients with mycosis fungoides (MF).Methods Totally,15 paraffin-embedded tissue specimens,14 fresh tissue specimens and 18 whole blood specimens were obtained from 28 patients with MF,and subjected to DNA extraction.BIOMED-2 multiplex PCR tubes TCRγ (A+B) were used for the analysis of TCRγgene rearrangements.Data were processed by SPSS 13.0 software,and statistical analysis was done by chi-square test and Fisher's exact probability test.Results TCR-γ gene rearrangements were detected in 3 paraffin-embedded tissue specimens,11 fresh tissue specimens and 12 blood specimens,with significant differences in the detection rate between the three samples (x2 =13.047,P ＜ 0.01).The fresh tissue samples showed a significantly higher detection rate than the paraffin-embedded tissue samples (X2 =12.523,P ＜ 0.01).The detection rate of TCRγgene rearrangements was 3/6 in paraffin-embedded tissue samples collected in 2011,significantly higher than that in the other 9 paraffin-embedded tissue samples collected before 2011 (Fisher's exact probability test,P =0.044),but similar to that in 14 fresh tissue specimens (12/14,Fisher's exact probability test,P =0.044).Decreased detection rate of TCRγ gene rearrangements was observed in blood samples compared with fresh tissue specimens,but no statistical difference was observed between the two types of specimens (x2 =2.358,P ＞ 0.05).Conclusions BIOMED-2 multiplex PCR tubes TCRγ(A+B) are suitable for the detection of clonal rearrangements of TCRγgene in different types of specimens,especially in fresh tissue specimens,from patients with MF.

Objective To analyze the effect of different treatment conditions on cells synchronization in G0/G1 phase to get the best con-dition, and to explore its effect on neural differentiation of bone marrow mesenchymal stem cells (BMSCs) induced by basic fibroblast growth factor (bFGF). Methods BMSCs were isolated and cultured in 5%, 1%, 0.5%, 0.1%, 0 fetal bovine serum (FBS) respectively, for 24 hours and 48 hours. After PI staining, cell cycle proportions of each phase were detected by flow cytometry, and were compared with the normal group (10%FBS). After the optimal treatment condition was got, 20 ng/ml bFGF was added into synchronization group and unsyn-chronization group 3 days and 7 days, respectively. The expression of Nestin and Tuj-1 were detected with immunofluorescence. Results Adult rat BMSCs were isolated from bone marrow and cultured, after passage, the cells were with long spindle shape. Compared with the normal group, the cell proportion of G1/G0 phase increased under different treatments, peaked with (94.274 ± 0.468)%under 1%FBS, 48 hours (F=39.91, P<0.001). After bFGF induction for 3 days, the Nestin+cell number was higher in the synchronization group than in the un-synchronization group [(80.3 ± 2.4)%vs. (12.1 ± 1.5)%] (F=28.25, P<0.001). After bFGF induction for 7 days, the Tuj-1+cell number was higher in the synchronization group than in the unsynchronization group [(74.8±3.2%)%vs. (19.3±2.5)%] (F=17.95, P<0.001). Conclusion 1%FBS, 48 hours is the optimal condition to BMSCs synchronization in G0/G1 phase, which can promote the neural differentiation of BM-SCs.

Objective To comparative study the effect on colorectal cleansing of CT colonography with gulping down 10 mg bisacodyl before or 1 h after oral taking 2 liter polyethylene glycol .Methods Forty participants with informed consent were appor‐tioned to group A ,group B randomly ,20 cases in each group .On the day before CT colonography ,participants in group A oral took 20 mL of 40% W/V barium sulfate prior to 3 mealtime ,and 20 mL of 60% diatrizoate meglumine diluted in 250 mL of water after supper ,then gulped down 10 mg bisacodyl enteric‐coated tablets 1 hour before oral taking 2 liter polyethylene glycol electrolyte so‐lution .Participants in group B were the same as that in group A ,with the exception of gulping down 10 mg bisacodyl enteric‐coated tablets 1 hour after oral taking 2 liter polyethylene glycol electrolyte solution .Cleansing efficacy of stool and fluid ,and attenuation value of remainder fluid between the two groups were analyzed statistically .Results In group A ,score of cleansing efficacy of stool (1 .96 ± 0 .11) was lower than that in group B (2 .01 ± 0 .12) ,segments with good cleansing efficacy of stool (87/120 segments , 72 .50% ) was higher than that in group B (83/120 segments ,69 .17% ) ,the difference was not statistically significant (P>0 .05) .In group A ,score of cleansing efficacy of fluid (1 .50 ± 0 .06) was lower than that in group B (1 .53 ± 0 .06) ,segments with good cleansing efficacy of fluid(113/120 segments ,94 .17% ) was higher than that in group B (111/120 segments ,92 .50% ) ,the differ‐ence was not statistically significant (P>0 .05) .Attenuation value of remainder fluid [(729 ± 29)HU ] in group A was higher than that in group B[(653 ± 25)HU] ,the difference was statistically significant(P<0 .05) .Conclusion Gulping down 10 mg Bisacodyl before or after oral taking 2 liter polyethylene glycol has no effect on cleansing of stool and fluid ,with good cleansing efficacy .The former has better cleansing efficacy of fluid ,is beneficial to detecting polyps for CT colonography .

Objective To observe the effects of neurotrophin 3 (NT3)-chitosan on motor function, and proliferation and differentiation of the neural stem cells (NSCs) in the injury area and subventricular zone (SVZ) in rats with motor cortex injury. Methods Sixty-five Wistar rats were divided into control group (n=7), injury group (n=29) and NT3-chitosan group (n=29). The motor cortex was aspirated and re-moved as cerebral injury model. NT3-chitosan was immediately implanted into the injured area after operation, and the control group re-ceived no intervention. Pellet reaching test was performed to detect the recovery of the forelimb function, HE staining was used to observe the lesion cavity size, and immunofluorescence staining was used to observe the proliferation and differentiation of NSCs 3 days, 7 days, 14 days, 1 month, 2 months and 3 months after operation. Results The grasp success rate was higher (F>6.00, P≤0.05), and the lesion cavity size was significantly smaller (F>629.5, P<0.001) in the NT3-chitosan group than in the injury group. In the NSCs differentiation experi-ment, the number of BrdU cells at all the time points was significantly higher in the NT3-chitosan group than in the injury group (F>171.43, P<0.001). In the NSCs proliferation experiment, the number of BrdU positive cells was still significantly higher in the NT3-chitosan group than in the control group and in the injury group (F>155.06, P<0.001), the number of Dcx positive cells was significantly higher in the NT3-chitosan group than in the injury group (F=62.367, P<0.001), and the number of BrdU/Dcx positive cells was significantly higher in the NT3-chitosan group than in the control group (F=33.527, P<0.001). Conclusion NT3-chitosan could activate NSCs in the SVZ, and pro-mote endogenous neurogenesis and forelimb function recovery in rats after motor cortex injury.

Objective To activate and amplify γδT cells with low molecular peptide antigen of Mycobacterium tuberculosis low molecular peptide antigen (Mtb-Ag),and to investigate the expression of CD69 molecules on γδT cellular surface.Methods Healthy human peripheral blood mononuclear cells (PBMC) were obtained and separated,then positive cells were isolated by immuno-magnetic beads selection,and the proportion of γδT cells in the PBMCs was detected by fluorescent monoclonal TCR γδT-PE staining and flow cytometry.Expression of CD69 molecules in γδT cells was detected by γδ-PE/CD69 FITC double staining.Results The proportion of γδT cells was (4.9±1.85)％ in freshly obtained PBMC,(69.2±6.57)％ after 10 d of Mtb-Ag activation,and (99.3±8.92)％ after immuno-magnetic beads selection.The expression of CD69 molecules in γδT cells reached the peak (75.2％) at 24 h after initial Mtb-Ag stimulation,and reached the peak (72.0％) at 6 h after second stimulation.Conclusions MtbAg can specifically stimulate the proliferation of γδT cells in the PBMC.Both its initial and the second stimulation can specifically activate γδT cells.