ABSTRACT
Objective: To screen for an optimized siRNA sequence targeting rat Toll-like receptor 4 (TLR4) in vitro. Methods: The full length gene of rat TLR4 was cloned and inserted into pEGFP-C1 plasmid to construct pEGFP-rTLR4. Three pairs of siRNAs targeting rTLR4 were chemically synthesized and were co-transfected with pEGFP-rTLR4 into HEK-293 cells via Lipofectamine2000. Cells were also co-transfected with siRNA targeting EGFP and negative control siRNA. The expression of EGFP was observed under inverted fluorescene microscope and flow cytometry. Results: Compared with the negative control group, 3 pairs of siRNAs targeting TLR4 and one pair of siRNA targeting EGFP significantly suppressed the EGFP expression (P<0.05); the inhibitory effect of siRNA2 (gene sequence: 5′-GTC TCA GAT ATC TAG ATC T-3′, 19 bp, 1 352-1 370) was the strongest one, with an interference efficiency over 75%. Conclusion: We have successfully obtained the siRNA sequence targeting TLR4 mRNA, which can efficiently suppress the expression of rat TLR4 mRNA in vitro.
ABSTRACT
Objective: To prepare a simple, practical fiberoptic bronchoscope (FB) training simulator, and to evaluate its training efficacy. Methods: A box apparatus was designed for basic and advanced FB training; the box consisted of five boards with holes in it. The holes were aligned according to the entries of bronchus to pass the FB. Assistant materials could be placed in the box to simulate foreign bodies due to diseases, which was aimed to help the training of sophisticated skills including tracheal tube intubation, biopsy, brushing and bronchoalveolar lavage. To testify the training efficacy, forty volunteer residents who had never manipulated a FB were randomly assigned into two groups: one group was trained using the prepared training box (Group TB, n = 20) and the other group was trained using video-based technique (Group C, n = 20). The trainees of the two groups received an intubation examination using an advanced patient simulator, and the periods of intubation was compared between the two groups. Results: Flexible fiberoptic bronchoscope could smoothly pass through the holes in the box; the photos of the holes under bronchoscopy well simulated the normal images. All residents completed the training and examination. The mean duration of intubation was (55. 7 ± 10. 2) s in Group TB and (69. 3 ± 8. 9) s in Group C (P<0. 001). Conclusion: The present training box is simple and practical, and it can be used for advanced training of FB, with obvious training efficacy.
ABSTRACT
Objective To examine the changes in the expression of voltage-gated sodium channel?subunit mRNA in the dorsal root ganglion(DRG)and the role it plays in the neuropathie pain.Methods Thirty- two male SD rats weighing 250-400g were randomly divided into 2 groups:groupⅠneuropathic pain(SNL,n= 20)and groupⅡsham operation(n=12).Neuropathic pain was produced by ligation of right sciatic nerve according to Seltzer.Paw withdrawal latency to noxious thermal(PWHL)and mechanical(PWML)stimulation were measured before(baseline)and 1,2,3,5,8,11,14,28 day after sciatic nerve ligation(SNL).DRG at L_(4,5) was isolated on the 14th day after SNL in 8 SNL and 4 sham-operated animals for determination of sodium channel?subunit mRNA expression(by in-situ hybridization).Results PWHL and PWML were significantly decreased on the 2nd-28th day after SNL as compared to the baseline in SNL group.There was no significant difference in?_1 subunit mRNA expression between the 2 groups.The?_2 subunit mRNA expression in DRG was hardly detectable.The?_3 subunit mRNA expression in DRG on the operated side was significantly higher in SNL group than in sham-operation group.Conclusion The up-regulation of sodium channel?_3 subunit mRNA expression in DRG may play an important role in neuropathic pain.