[Effects of retinopathy on visual function in type 2 diabetes mellitus].

Objective: To study the influence of the severity of diabetic retinopathy (DR) on the visual function of patients with type 2 diabetes, to provide scientific basis for the early prevention and control of DR. Methods: This study was designed as a cross-sectional study, recruiting already-diagnosed type 2 diabetes patients in four community health service centers in Guizhou Province between February and September 2022. Employing the Chinese version of the Visual Function Index-14 (VF-14), assess the participants' near vision, visual adaptation, subjective visual perception, and stereo vision, with higher scores indicating poorer visual function. Categorize the severity of each eye's damage into no diabetic retinopathy (DR), mild non-proliferative diabetic retinopathy (NPDR), moderate NPDR, severe NPDR, and proliferative diabetic retinopathy (PDR), and use a 5-level DR grading system to evaluate the overall severity of diabetic retinopathy in both eyes. Employing linear regression analysis to investigate the linear relationship between DR and visual function index. Local weighted regression evaluates the nonlinear relationship between the DR composite score and the scores of visual function, with a steeper slope indicating poorer visual function for that level. Results: A total of 542 patients with type 2 diabetes were investigated, including 244 (45.02%) males, 298 (54.98%) females, and 162 (29.89%) patients with DR. After adjusting for confounders, compared with those without DR, patients with binocular DR Had overall scores (ß=0.136, P=0.003), near vision (ß=0.163, P<0.001), visual adaptation (ß=0.092, P=0.042), subjective vision (ß=0.120, P=0.009) and stereo vision (ß=0.094, P=0.044) were higher than those without DR. There were no differences in visual functions between DR And monocular DR. The local weighted regression curve showed that near vision (slope: 23.78) and overall score (slope: 58.37) increased sharply from mild to moderate NPDR in both eyes. Visual adaptation (slope: 5.37, 7.72), subjective vision (slope: 6.53, 7.93), stereovision (slope: 0.74, 0.91) increased slowly in mild to moderate NPDR in both eyes and in moderate to severe NPDR/PDR in both eyes. Conclusion: Binocular DR is associated with impaired visual function, but there is no difference between monocular DR And non-DR visual function. The early damage of DR To visual function is mainly manifested in near vision. In the prevention and control of DR, more attention should be paid to visual function, especially the change of near vision, and retinal damage should not be assessed solely by visual status.

[Effect of RNA interference targeting neuropilin-2 gene on proliferation and apoptosis of colon cancer cell line HCT-8].

Objective: To investigate the effects of down-regulation of expression of neuropilin-2 (NRP-2) by RNA interference (RNAi) technique on proliferation and apoptosis of HCT-8 colon cancer cells. Methods: NRP2-siRNA and negative control (NControl)-siRNA were transferred into HCT-8 colon cancer cells by liposomes (lip2000) as transfection group and negative control group, and phosphate buffered solution (PBS) was added as blank control group. Quantitative reverse transcription PCR (RT-qPCR) and Western blot were used to detect the transfection effect. The proliferation of cells in the three groups was examined by cell counting kit (CCK) assay, colony-forming unit assay and Ki-67 protein staining assay, respectively. Moreover, the apoptosis of cells in the three groups was determined by acridine orange/propranidine iodide (AO/PI) staining method. Results: The results of RT-qPCR and Western blot showed that the relative expression of NRP-2 mRNA and the content of NRP-2 protein in the transfer group decreased (0.46±0.05 vs 0.99±0.05 and 1.00±0.06; 1.04±0.06 vs 1.73±0.09 and 1.65±0.11) (all P<0.05). The results of CCK-8 demonstrated that the optical density of transfection group was significantly lower than that of the negative control group and the blank control group(24 h: 0.53±0.04 vs 0.82±0.07 and 0.87±0.07; 48 h: 0.54±0.05 vs 1.00±0.09 and 1.17±0.05; 72 h: 0.75±0.05 vs 1.31±0.13 and 1.50±0.03; 96 h:1.05±0.04 vs 1.46±0.09 and 1.86±0.06) (all P<0.05). The results of colony-forming unit assay indicated that the proliferation ability of the cells in the transfer group was significantly lower than that in the other two groups (134.67±8.74 vs 245.33±19.14 and 300.33±14.01, P<0.05). The results of Ki-67 protein staining assay showed that compared with the negative control group and blank control group, the expression of Ki-67 protein was significantly decreased in the transfection group (5.93±0.22 vs 8.36±0.09 and 8.70±0.21, P<0.05). The results of AO/PI assay revealed that the ratio of apoptotic cells to living cells in the transfer group was significantly higher than that in the other two groups (0.43±0.07 vs 0.14±0.04 and 0.11±0.04, P<0.05). Conclusion: The proliferation ability of HCT-8 colon cancer cells decreases, and the apoptosis ability increases by decreasing the expression of NRP-2.

The effect of neuropilin-1 silencing on the transforming growth factor-ß1-mediated epithelial-mesenchymal transition of colon cancer SW480 cells and its effect on the proliferation and migration of colon cancer cells.

This study aimed to investigate the effect of neuropilin-1 (NRP-1) silencing on epithelial-mesenchymal transformation (EMT) mediated by transforming growth factor-ß1 (TGF-ß1) and on the proliferation and migration of colon cancer SW480 cells. After transfection of small interfering ribonucleic acid (siRNA)-NRP-1 into colon cancer SW480 cells, the messenger RNA (mRNA) and protein expression levels of NRP-1 were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Four EMT models were induced using 0, 2, 5, and 10 ng/mL TGF-ß1, respectively. Cell proliferation was detected using Cell Counting Kit-8, and the protein levels of EMT markers E-cadherin and vimentin were detected using Western blot. EMT was induced in the transfected SW480 cells using TGF-ß1, after which four groups were created: a negative control group (siRNA-Ncontrol), a transfection group (siRNA-NRP-1), an induction group (TGF-ß1), and a transfection + induction group (siRNA-NRP-1+TGF-ß1). Western blot was then used to detect the expression of E-cadherin and vimentin, and cell proliferation and migration were detected using cell counting kit-8 (CCK-8) and scratch assay. After transfection with siRNA-NRP-1, the mRNA and protein expression levels of SW480 cells were significantly decreased (P<0.05). After 48 hours of induction with 10 ng/mL TGF-ß1, cell proliferation was obvious, E-cadherin expression decreased, and vimentin expression significantly increased (P<0.05), indicating that EMT had been successfully induced compared with the induction group, the transfection + induction group had significantly increased E-cadherin expression after corresponding treatments (including transfection and induction alone) (P<0.05), and the proliferation and migration of colon cancer cells decreased (P<0.05). In conclusion: silencing, NRP-1 in colon cancer SW480 cells can partially reverse TGF-ß1-mediated EMT, reduce the proliferation activity of colon cancer cells, and slow their migration ability. Therefore, NRP-1 may become a new target for the treatment of colon cancer.

First Report of Broad bean wilt virus 2 Infecting Bupleurum chinense in China.

Samples of the medicinal plant, Bupleurum chinense DC., were collected in October 2007 from the garden of the Institute of Medicinal Plant Development in Beijing. Partial fragments of the genomic RNA1 and RNA2 of Broad bean wilt virus 2 (BBWV-2) were obtained from the root cDNA library of these plants. Sequence analyses showed the 1,896-nt RNA1 fragment (GenBank No. FJ485684) encoding a portion of the RNA-dependent RNA polymerase (RdRp) and the 2,017-nt RNA2 fragment (No. FJ485685) encoding 612 amino acids of the complete large (LCP) and small coat protein (SCP), respectively. The amino acid identities of LCP and SCP were 90.8 to 96.7% compared with sequences of other BBWV-2 isolates deposited in the GenBank with the highest homology to Japanese IP (No. AB018698) and the lowest to Japanese 1-2 (No. AB018701). This strongly suggests that the B. chinense plants utilized for cDNA library construction were infected by what appears to be an isolate of BBWV-2. Seeds from the same batch were sown again in the same garden in May 2008. In August 2008, approximately 30% of these plants showed mosaic, distortion, and stunting. Reverse transcription (RT)-PCR amplicons were obtained from eight symptomatic plants using a pair of conserved primers for specific detection of viruses within the Fabavirus genus (2). A symptomless plant tested negative by RT-PCR. The same single 391-bp amplicon of RNA1 (No. FJ485686) obtained from five of those eight symptomatic plants were cloned and sequenced. Sequence comparison with the corresponding sequences of other BBWV-2 isolates showed that the sequenced isolate was most closely related to B935-a Chinese faba bean isolate (No. AF149425). Crude sap of one diseased B. chinense plant was used for mechanical inoculation to Chenopodium amaranticolor Coste & Reyn. Chlorotic local lesions were observed on inoculated leaves 5 days after inoculation, and subsequently, systemic mottle and malformed symptoms appeared on the upper leaves. Twelve plants were inoculated and all plants showed symptoms of virus infection. RT-PCR tests of inoculated indicator plants showing local lesions confirmed the presence of BBWV-2. To date, Clover yellow vein virus and Lettuce mosaic virus have been isolated from the genus Bupleurum (B. griffithii hort. and B. falcatum L. sensu lato) in Japan and Israel, respectively (1,3). Furthermore, to our knowledge, no genomic sequence of BBWV-2 naturally infecting plants in the family Umbelliferae/Apiaceae has been reported. Therefore, this is the first report of BBWV-2 on B. chinense (Umbelliferae/Apiaceae), which was designated as a BC isolate of BBWV-2. In China, BBWV-2 was reported to be infecting and causing heavy losses to many plant species mostly belonging to the family Leguminosae (4). B. chinense is a commonly used bulk medicinal plant mainly cultivated in Hebei, Sichuan, Gansu, and Shanxi provinces in China for decoction pieces and extracts of its dried roots, which are also exported to Japan, Korea, and Southeast Asia. These results demonstrate the need for further assessment of BBWV-2 incidence and the losses it may cause. References: (1) J. Cohen et al. Phytoparasitica 30:88, 2002. (2) R. M. Ferrer et al. J. Virol. Methods 144:156, 2007. (3) H. Yamamoto. Jpn. J. Phytopathol. 69:420, 2003. (4) X. P. Zhou et al. Acta Phytopathol. Sin. 26:347, 1996.

A calmodulin-binding peptide of caldesmon.

Caldesmon is a major actin-binding protein identified in smooth muscle and many non-muscle cells. It also interacts with calmodulin and a number of other acidic proteins. We have shown previously that the polypeptide stretch from Val629 to Ser666 near the C terminus contains a calmodulin binding site (Wang, C.-L. A., Wang, L.-W. C., Xu, S., Lu, R. C., Saavedra-Alanis, V., and Bryan, J. (1991) J. Biol. Chem. 266, 9166-9172). On the other hand, Bartegi et al. (Bartegi, A., Fattoum, A., Derancourt, J., and Kassab, R. (1990) J. Biol. Chem. 265, 15231-15238) reported a cyanogen bromide fragment beginning at Trp659 which is also capable of binding both calmodulin and actin. A comparison of the overlapping sequence between these two peptides suggests that this calmodulin binding site is localized in a 7-residue segment, 659Trp-Glu-Lys-Gly-Asn-Val-Phe665. We have chemically synthesized an 18-residue peptide (GS17C, from Gly651 to Ser667 with an added cysteine at the C terminus) that contains this segment. This peptide was purified by high performance liquid chromatography and labeled with fluorescent probes at the terminal cysteine residue. We found that GS17C indeed binds calmodulin in a Ca(2+)-dependent manner (Kd = 8 x 10(-7) M) and appears to compete with caldesmon. Interestingly, this synthetic peptide also co-sediments with F-actin, binding to actin being displaceable by calmodulin, as in the case of the native caldesmon. But GS17C does not have any effect on the actomyosin ATPase activity, indicating that this peptide segment does not contain the inhibitory region.