OBJECTIVETo study the role of integrin-linked kinase (ILK) on the proliferation and differentiation of human fibroblast in hypertrophic scar and its effect on the scar formation.

METHODSThe human scar fibroblasts were isolated and cultured in vitro. The cells were divided into 4 groups. (1) control group: only contains DMEM; (2) jetPRIME group: DMEM with 200 microl jetPRIME buffer and 4 microl jetPRIME; (3) ILK siRNA group: DMEM and ILK siRNA; (4) ILK cDNA group: DMEM and ILK cDNA. The cell proliferation was detected by XTT assay and the mRNA and protein expressions of ILK and alpha-SMA were detected by Real-time qPCR and Western blot.

RESULTS(1) XTT results showed that the cellular proliferation level after 48 h in four groups were 0.820 +/- 0.065, 0.873 +/- 0.041, 0.554 +/- 0.013 and 1.296 +/- 0.094, respectively. The cellular proliferation curve showed that the cellular proliferation level was very flat in ILK siRNA group while the cellular proliferation level gradually increased from 12 h. 48 h after transfection, the cellular proliferation level in ILK siRNA group was significant lower than those in other groups (P value were 0.021, 0.034, 0), while the cellular proliferation level in ILK cDNA group was the highest among all 4 groups (P value were 0.017, 0.009, 0). (2) The Real-time qPCR showed that the expressions of ILK mRNA and alpha-SMA mRNA were 0.693 +/- 0.412 and 0.422 +/- 0.037 in control group, were 0.621 +/- 0.183 and 0.388 +/- 0.005 in jetPRIME group, were 0.052 +/- 0.019 and 0.073 +/- 0.023 in ILK siRNA group, were 240.193 +/- 35.170 and 138.056 +/- 24.060 in ILK cDNA group. The expressions of ILK mRNA and alpha-SMA mRNA in ILK siRNA group were significantly lower than those in other three groups (P < 0.05). And the expressions of ILK mRNA and alpha-SMA mRNA in ILK cDNA group were significantly higher than those in other three groups (P < 0.05). (3) The Western blot also showed that the expression of ILK and alpha-SMA proteins were decreased in ILK siRNA group and increased in ILK cDNA group.

CONCLUSIONILK may promote the proliferation and differentiation of human scar fibroblast. It may play an important role in scar formation and contracture.

Actins ; metabolism ; Adolescent ; Adult ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; Female ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; pharmacology ; RNA, Messenger ; genetics ; Transfection ; Young Adult

Objective To investigate the role of three-dimensional scaffolds seeded with NeuroD1-modified neural stem cells (NSCs) for repair of spinal cord injury in rats.Methods A new three-dimensional bio-printer was used to make bionic spinal cord scaffolds.NSCs are transduced with retrovirus vectors encoding NeuroD1 to express transgenes in high levels.Forty healthy female SD rats were divided into control group,scaffold group,scaffold + NSCs-green fluorescent protein (GFP) group and scaffold + NSCs-NeuroD1 group with 10 rats per group,according to the random number table.Spinal cord injury in rats was induced using the electric controlled cortical impactor.A week later,the control group was excised 3 mm spinal cord at the injury site under microscope.RT-PCR was used to confirm the construction of NeuroD1 overexpressing NSCs.Survival and differentiation of transplanted NSCs were detected with fluorescent staining.Rat neurological motor function was evaluated with BBB score at postoperative 1,2,4,6 and 8 weeks.Rat electrophysiological changes were observed by monitoring motion evoked potential and sensory evoked potential at 8 weeks.Results RT-PCR results confirmed the successful reconstruction of NeuroD1-overexpressing NSCs.BBB score in scaffold + NSCs-NeuroD1 group was the highest and had significant differences compared to other three groups (P ＜ 0.05).Electrophysiological results showed the motor and sensory in scaffold + NSCs-NeuroD1 group had the shortest latencies and highest amplitudes,which revealed significant differences compared to other three groups (P ＜0.05).Immunofluorescence staining showed GFP cells in scaffold + NSCs-NeuroD1 group at 8 weeks,which differentiated into neurons and astrocytes.GAP-43 was positively stained,and myelin formation was detected.Conclusion Three-dimensional scaffolds seeded with NeuroD1-modified NSCs can promote nerve loop reconstruction in spinal cord injury rats,and accelerate recovery of motor and sensory function.

Objective To compare the effects of different support method on eccentric control angle and muscle strength of knee joints in 10-week Nordic hamstring exercise.Methods Twenty-eight college male football players were randomly divided into hand support group (HS group, n=14) and non-hand support group (NHS group, n=14) for 10 weeks. The eccentric control angle and eccentric muscle strength of knee joints were tested before and after the intervention.Results After 10 weeks of Nordic hamstring training, the eccentric control flexion angle of knee joint in HS group and NHS group was significantly decreased by 8° and 10°, and the knee joint angle in NHS group was 12° lower than that in HS group (P<0.05); the peak torque of hamstring and quadriceps femoris in both HS group and NHS group was significantly increased, and the peak torque of hamstring centrifugal motion at 60°/s and 120°/s in NHS group was 16 and 13 N·m higher than that in HS group, respectively (P<0.05).Conclusions NHS group is better than HS group in improving knee flexion angle and centrifugal muscle force. It is suggested that Nordic hamstring training should be taken as one of the daily training tasks for football players to prevent hamstring injury.