Rab GTPases serve as master regulators of vesicular membrane transport on both the exo-and endocytic pathways. Though there are many reports on Rab proteins, the function of these small proteins still remain in speculation. And no report has ever clarified the character of human Rab26. Here it was reported that a novel Rab protein Rab26 is membranous organelle related and in volved inendocytosis of HeLa cells. By using RT-PCR method a novel Rab26 cDNA full-length cDNA of Rab26 that is 1656 bp was identified.The cDNA sequence that at 1197 is 'A' other than 'G', while 'C' at 956 substitutes for 'T', and has 'GCC' insertion at 48 to 50 compared with published sequences. The complete open reading frame (ORF) is 771 bp in length encoding 256-residue protein with a calculated molecular mass of 27.9 ku (GenBank accession No.AY646153), rather than a shorter one with 190-amino acid residue as reported previously. GFP labeled full-length Rab26 expression showed that Rab26 was mainly sublocated in membranous organelles and could enhance endocytosis which means could took PE labeled protein as an endocytic tracer. RT-PCR analysis showed Rab26 was detected to express in several kinds of adenocarcinoma cell lines such as Acc2, AccM, SPC-A1 and HeLa cell lines, which indicated that Rab26 expression might be associated with some carcinomas.

Objective:To investigate the effects of applying Stent Boost Subtract (SBS) technique during percutaneous coronary intervention in patients with complex coronary lesions.Methods:200 patients with coronary artery disease (CAD) who hospitalized in the department of cardiology in Guangzhou First People′s Hospital from June 2018 to June 2020 were enrolled. The coronary lesions of all patients were corresponding to B2 or C type suggested by American Heart Association (AHA)/American College of Cardiology (ACC) according to coronary angiography and treated with percutaneous coronary intervention (PCI). Patients were randomly divided into SBS group ( n=82, SBS technique was applied during PCI) and IVUS group [ n=118, intravascular ultrasound (IVUS) was applied during PCI]. After stent implantation, quantitative coronary angiography (QCA) automatic analysis system was used to measure the related parameters of stent diameter (including the minimum, maximum and mean value of stent diameter) and calculate the stent eccentricity index. During PCI, stent eccentricity index, post-stent expansion, poorly positioned stent with open lesions, failure of overlapping stent with long lesions, mean cumulative dose (CD), product of total dose area (DAP), X-ray time, operation time and operation cost of each PCI were recorded in the two groups. Patients were followed up for 18 months after PCI, and the occurrence of adverse cardiovascular events (MACE) was recorded during the follow-up period, and the cumulative survival rate without MACE was compared between the two groups. Results:There were no statistically significant differences between the two groups in stent eccentricity index, proportion of guided stent expansion, proportion of poorly positioned stent with open lesions, proportion of stent failure to overlap, with statistically significant difference[(0.12±0.04) vs (0.10±0.03); 80.49% vs 85.49%; 2.44% vs 2.54%; 1.22% vs 2.54%, all P>0.05]. There were no significant differences in CD, X-ray time and DAP in SBS group compared with IVUS group [(1 394.18±42.29)Gy/cm 2 vs (1 391.82±45.06)Gy/cm 2; (18.79±3.01)min vs (18.95±3.12)min, (100.24±5.70)Gy/cm 2 vs (99.47±5.93)Gy/cm 2; all P>0.05]. The operation time in SBS group was shorter than that in IVUS group [(70.91±6.51)min vs (73.89±8.95)min, P<0.05], and the operation cost was less than that in IVUS group [(2.98±0.86)ten thousand yuan vs (3.85±0.81)ten thousand yuan, P<0.05]. After 18 months of follow-up after PCI, Kaplan-Meier survival analysis showed that there was no significant difference in MACE event-free survival between SBS group and IVUS group (91.46% vs 94.07%, Log Rank=0.480, P=0.489). Conclusions:SBS is a kind of convenient and effective technique in guiding PCI in patients with complex coronary lesions without increasing operation time and radiation dose, which can achieve the same effect as IVUS guidance.

ObjectiveTo investigate the effects of microRNA (miRNA, miR)-887-3p on the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells and its underlying molecular mechanism. MethodsAnnulus fibrosus tissues were obtained from 8-week-old SPF-grade SD male rats, centrifuged to prepare and identify annulus fibrosus cells. Rats in the experiment were randomly divided into four groups: a Normal group consisting of primary annulus fibrosus cells without any treatment; a Control group treated with 10 ng/mL interleukin-1β (IL-1β) for 24 hours to establish a degenerative cell model; an interference group (miR-887-3p inhibitor) transfected with miR-887-3p inhibitor using Lipo3000 based on the Control group; and an overexpression group (miR-887-3p mimics) transfected with miR-887-3p mimics using Lipo3000 based on the Control group. CCK-8 assay was used to assess cell viability; flow cytometry was used to measure cell apoptosis rates; real-time fluorescence quantitative PCR (qPCR) was used to detect the expression levels of miR-887-3p and murine double minute 4 (MDM4) mRNA; Western blotting was used to measure the protein expression levels of MDM4, Bcl-2, and Caspase-3. ResultsImmunofluorescence staining of isolated and cultured cells revealed a Collagen I positive rate of over 90% in rat intervertebral disc annulus fibrosus cells, indicating a cell purity level greater than 90%. Real-time fluorescence qPCR results showed that after establishing an annulus fibrosus degenerative cell model using IL-1β, the expression level of miR-887-3p significantly increased compared to the Normal group (P<0.001). Compared to the Control group, transfection with miR-887-3p inhibitor resulted in a significant decrease in its expression level (P<0.001). The CCK-8 assay showed that compared to the Normal group, cell viability significantly decreased in the Control group (P<0.001). Compared to the Control group, cell proliferation ability significantly increased after miR-887-3p inhibition, and significantly decreased after overexpression of miR-887-3p. Flow cytometry results revealed that compared to the Normal group, the apoptosis rate in the Control group significantly increased (P<0.001). Compared to the Control group, the cell apoptosis rate significantly decreased in the miR-887-3p interference group (P<0.001) and increased in the overexpression group (P<0.001). Western blotting analysis showed that compared to the Normal group, Bcl-2 expression level significantly decreased (P<0.001) and Caspase-3 expression level significantly increased (P<0.001) in the Control group. Compared to the Control group, Bcl-2 and MDM4 expression levels significantly increased (P<0.01), and Caspase-3 expression level significantly decreased (P<0.01) in the miR-887-3p interference group; whereas in the overexpression group, Bcl‑2 and MDM4 expression levels significantly decreased (P<0.05), and Caspase-3 levels significantly increased (P<0.05). Real-time fluorescence qPCR and protein immunoblotting results showed that after interfering with miR-887-3p, the expression of MDM4 protein and mRNA increased (P<0.001); after overexpressing miR-887-3p, their expression decreased (protein, P<0.01; mRNA, P<0.001). ConclusionMiR-887-3p may modulate the cell proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells by regulating MDM4 expression, thereby influencing the development and progression of disc degeneration.