DataSheet1_The Changes in Drainage Systems of Weihe Basin and Sanmenxia Basin Since Late Pliocene Give New Insights Into the Evolution of the Yellow River.docx

The formation of the Yellow River involved the draining of a series of ancestral local lakes along their course, substantially changing the regional, geomorphic, and paleoenvironmental evolution. However, the evolution of the Weihe-Sanmenxia Basin section of the Yellow River remains indistinct as previous studies regard the Weihe and Sanmenxia Basin as one integral basin of the Late Cenozoic. Here, we present the detrital zircon age spectra from the Pliocene-Pleistocene Sanmen Formation to clarify the drainage system evolution of the two basins since the Late Pliocene. The results reveal that these two basins belonged to different drainage systems in the Late Pliocene because no sediments from the marginal mountains of the Weihe Basin accumulated in the Sanmenxia Basin. At 2.8/2.6 Ma, the currents presented at the edge of the basins and transported the sediment of east Hua Mountain into the Sanmenxia Basin, where it was deposited. This integration likely leads to a mismatch between the deposition and regional paleoclimate in previous studies. At ∼1.0 Ma, the Sanmenxia Gorge was traversed and the Yellow River finally formed, depositing Jinshaan Gorge sediment into the Sanmenxia Basin and lower reaches of the Yellow River.</p

Primer sequences for real-time PCR.

<p>Primer sequences for real-time PCR.</p

DataSheet2_The Changes in Drainage Systems of Weihe Basin and Sanmenxia Basin Since Late Pliocene Give New Insights Into the Evolution of the Yellow River.xlsx

The formation of the Yellow River involved the draining of a series of ancestral local lakes along their course, substantially changing the regional, geomorphic, and paleoenvironmental evolution. However, the evolution of the Weihe-Sanmenxia Basin section of the Yellow River remains indistinct as previous studies regard the Weihe and Sanmenxia Basin as one integral basin of the Late Cenozoic. Here, we present the detrital zircon age spectra from the Pliocene-Pleistocene Sanmen Formation to clarify the drainage system evolution of the two basins since the Late Pliocene. The results reveal that these two basins belonged to different drainage systems in the Late Pliocene because no sediments from the marginal mountains of the Weihe Basin accumulated in the Sanmenxia Basin. At 2.8/2.6 Ma, the currents presented at the edge of the basins and transported the sediment of east Hua Mountain into the Sanmenxia Basin, where it was deposited. This integration likely leads to a mismatch between the deposition and regional paleoclimate in previous studies. At ∼1.0 Ma, the Sanmenxia Gorge was traversed and the Yellow River finally formed, depositing Jinshaan Gorge sediment into the Sanmenxia Basin and lower reaches of the Yellow River.</p

BMP-2 Is Involved in Scleral Remodeling in Myopia Development

<div><p>The development of myopia is associated with scleral remodeling, but it is unclear which factors regulate this process. This study investigated bone morphogenetic protein-2 (BMP-2) expression in the sclera of guinea pigs with lens-induced myopia (LIM) and after recovery from myopia and evaluated the effect of BMP-2 on extracellular matrix (ECM) synthesis in human scleral fibroblasts (HSFs) cultured <i>in vitro</i>. Lens-induced myopia was brought about in two groups of guinea pigs (the lens-induced myopia and myopia recovery groups) by placing -4.00 D lenses on the right eye for three weeks. The left eye served as a contralateral control. In the recovery group, the lenses were removed after one week. The refractive power and axial length of the eyes were measured, and the BMP-2 expression levels in the sclera were measured. After three weeks, the lens-induced eyes acquired relative myopia in both groups of guinea pigs. Immunostaining of the eyeballs revealed significantly decreased BMP-2 expression in the posterior sclera of the myopic eyes compared to the contralateral eyes. One week after lens removal, BMP-2 expression recovered, and no differences were observed between the experimental and contralateral eyes in the recovery group. HSFs were cultured with BMP-2 or transforming growth factor-β1 (TGF-β1). Type I and type III collagen synthesis was significantly up-regulated following BMP-2 treatment in culture after one and two weeks, but the ratio of type III to type I collagen mRNA was not increased. Biosynthesis of glycosaminoglycan (GAG) and aggrecan was increased in HSFs treated with BMP-2. Some chondrogenesis-associated genes expression increased in HSFs treated with BMP-2. From this study, we concluded that BMP-2 is involved in scleral remodeling in the development and recovery of lens-induced myopia.</p></div

The effect of BMP-2 on chondrogenesis-related genes expression in HSFs.

<p>The mRNA level of SOX5 (A), SOX6 (B), SOX9 (C), RUNX2 (D), HAPLN (E), PTHR1 (F) was analyzed. HSFs were treated for 3 days, 1 week, and 2 weeks seperately. *: Denotes a significant difference relative to the control group, P< 0.05.</p

The effect of BMP-2 on type I collagen expression in HSFs.

<p>The expression of type III collagen in HSFs after 3 days (A), 1 week (B) and 2 weeks (C). Lane 1-Untreated control, Lane 2–10 ng/ml BMP-2, Lane 3–50 ng/ml BMP-2, Lane 4–100 ng/ml BMP-2, Lane 5–10 ng/ml TGF-β1. The data are expressed as the mean±SEM, N = 3, * p≤0.05 vs. control. Real-time PCR (E) also shows an increase in type III collagen after BMP-2 and TGF-β1 treatment (D). Immunocytochemistry (F) reveals the expression of type III collagen in HSFs after 7 days culture. The ratio of type III/I collagen mRNA increased significantly in HSFs treated with 50ng/BMP-2 for 3 days compared to control cells, but this ratio decreased and was not changed after 1 or 2 weeks of treatment (G). The data are expressed as the mean±SEM. N = 3, * p≤0.05 vs. control.</p

BMP-2 expression in myopia and recovery from myopia of guinea pigs.

<p>Immunofluorescence analysis of BMP-2 expression in the retina, RPE, choroid and sclera of guinea pigs. The staining in the myopic sclera of the LIM group was weaker than that of the control eyes (A). Changes in the expression of BMP-2 in the sclera of guinea pigs detected by western blot analysis (C,D). The data are expressed as the mean±SEM, * p≤0.05 vs. the contralateral eye. <i>INL</i> inner nuclear layer, <i>ONL</i> outer nuclear layer, <i>RPE</i> retinal pigmental epithelium, <i>chl</i> choroid, <i>Sc</i> sclera layer.</p

The effect of BMP-2 on GAG synthesis in HSFs.

<p>GAG synthesis was visualized by toluidine blue staining of HSFs in culture. GAG was observed after treatment with BMP-2 for one week but not in control cells. Original magnification: 40×.</p

The effect of BMP-2 on aggrecan expression in HSFs.

<p>Aggrecan mRNA level in HSFs was analyzed after 3 days, 1 week and 2 weeks (A). Immunocytochemistry shows an increase in aggrecan expression after 7 days (B). The data are expressed as the mean±SEM. N = 3, * p≤0.05 vs. Control.</p

The effect of BMP-2 on type II collagen expression in HSFs.

<p>Type II collagen synthesis in HSFs after 3 days (A), 1 week (B), 2 weeks (C) detected by western lot analysis (D). Lane 1- Untreated control, Lane 2–10 ng/ml BMP-2, Lane 3–50 ng/ml BMP-2, Lane 4–100 ng/ml BMP-2, Lane 5–10 ng/ml TGF-β1. The expression of type II collagen mRNA was analyzed using real-time PCR (E). Immunocytochemistry showed an increase in collagen II expression in HSFs after 7 days of treatment with BMP-2 and TGF-β1 in culture (F). The data are expressed as the mean±SEM. N = 3, * p≤0.05 vs. Control.</p