[Clinical management of thrombocytopenia in cirrhosis].

Thrombocytopenia is one of the common complications of cirrhotic patients, which can induce an increasing bleeding risk and closely correlate with bleeding following invasive procedures. Consequently, how to respond to thrombocytopenia is crucial for improving the prognosis of patients with cirrhosis. This article reviews the main mechanisms of cirrhosis concurrent with thrombocytopenia, as well as the corresponding clinical management strategies.

MicroRNA-28-5p regulates glioma cell proliferation, invasion and migration by targeting SphK1.

The article "MicroRNA-28-5p regulates glioma cell proliferation, invasion and migration by targeting SphK1, by H.-S. Chen, A.-Q. Lu, P.-Y. Yang, J. Liang, Y. Wei, Y.-W. Shang, Q. Li, published in Eur Rev Med Pharmacol Sci 2019; 23 (15): 6621-6628-DOI: 10.26355/eurrev_201908_18551-PMID: 31378904" has been withdrawn from the authors stating that "after our follow-up experiments and in-depth research, we found that the previous experimental data had some loopholes and deviations. After the experiment was improved, we found that some experimental data could not be repeated again. To avoid academic adverse effects, we ask the magazine to withdraw the manuscript". The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18551.

Simvastatin promotes osteogenic differentiation of mesenchymal stem cells in rat model of osteoporosis through BMP-2/Smads signaling pathway.

OBJECTIVE: By establishing osteoporosis (OP) model in rats, the specific regulatory effect of simvastatin on promoting the differentiation of mesenchymal stem cells (MSCs) into osteoblasts through the bone morphogenetic protein 2 (BMP-2)/Smads signaling pathway was investigated. MATERIALS AND METHODS: A total of 45 Sprague-Dawley rats were selected to establish the OP model by performing ovariectomy. The rats were divided into OP model group (OP group, n=15), 10-7 mmol/L simvastatin treatment group (SIM group, n=15), and normal control group (Control group, n=15). After the experimental period, the enzyme-linked immunosorbent assay (ELISA) was applied to observe the serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-1. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was adopted to detect the contents of the differentiation-associated genes [runt-related transcription factor 2 (RUNX2) and Osterix (Osx)]. Later, the bone marrow MSCs (BMSCs) were selected and divided into Control group, 10-7 mol/L simvastatin group (SIM group), and osteoinduction medium group (OM group). Cell morphology in each group was observed. The Cell Counting Kit-8 (CCK-8) was performed to determine the proliferation activity of BMSCs. ELISA was performed to measure the level of alkaline phosphatase (ALP). RT-PCR was conducted to examine the levels of key differentiation-associated gene RUNX2 and those in BMP-2/Smads pathway. Moreover, the Western blotting was adopted to analyze the expressions of RUNX2 and genes in BMP-2/Smads pathway. RESULTS: The serum levels of TNF-α, IL-6, and IL-1 in OP group were remarkably higher than those in the Control group, and their levels in the SIM group were close to those in the Control group. The elevated messenger ribonucleic acid (mRNA) levels of the key differentiation-associated factors RUNX2, osteoprotegerin (OPG), osteopontin (OPN), and Osx were observed in the SIM group. In vitro cell culture revealed that the cells were in a favorable growth status in the SIM group and OM group, mostly manifesting in fusiform or spindle shape, and proliferated rapidly. In addition, the ALP level notably increased in the two groups compared with that in the Control group (p<0.05). Both SIM group and OM group had evidently higher mRNA expression levels of RUNX2, OPG, OPN, and Osx than those in the Control group (p<0.05), consistent with the expression trends of the genes in BMP-2/Smads pathway. The Western blotting indicated that the expression levels of RUNX2 and genes in BMP-2/Smads pathway in the SIM group were significantly higher than those in the Control group. CONCLUSIONS: Simvastatin can promote the differentiation of MSCs into osteoblasts in the OP rat model through the BMP-2/Smads signaling pathway.

MicroRNA-28-5p regulates glioma cell proliferation, invasion and migration by targeting SphK1.

OBJECTIVE: MicroRNAs (miRNAs) are a conserved class of endogenous and short non-coding RNAs that post-transcriptionally regulate the expression of genes involved in diverse cellular processes. MiR-28-5p has been reported to be associated with several cancers, including human glioma. However, the roles of miR-28-5p in glioma development are poorly understood. MATERIALS AND METHODS: Sixteen human glioma tissues and paired adjacent normal tissues were acquired through the Gansu Provincial Hospital. We performed quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) to detect the miR-28-5p expression between 16 paired adjacent normal and glioma tissues, as well as the miR-28-5p expression between normal human astrocytes cells and five glioma cell lines. To examine the functional roles of the downregulated miR-28-5p in glioma, cell viability and colony formation assays were performed for the analysis of cell growth. We overexpressed miR-28-5p by transient transfection of miRNAs mimics and performed the transwell Matrigel invasion assay and transwell migration (without Matrigel) assay. To investigate the roles of miR-28-5p in SphK1 expression, Western blot and Real Time-Polymerase Chain Reaction assays were performed. RESULTS: In this work, we demonstrated that miR-28-5p is downregulated in glioma tissues compared to the adjacent normal tissues. Functional studies showed that miR-28-5p overexpression inhibited the cell viability, colony formation and proliferation; meanwhile, it induced the cell apoptosis. The transwell invasion assay indicated that miR-28-5p blocked the invasion and migration of glioma cells. SphK1 (Sphingosine kinase 1 antibody) is predicted as a targeted candidate of miR-28-5p. Then, the Luciferase reporter assay, Western blot and Real Time-Polymerase Chain Reaction (PCR) validated that miR-28-5p negatively regulated SphK1 expression by directly targeting its 3'untranslated regions (3'UTR) in U87 cells. Furthermore, rescue assay suggested that overexpression of SphK1 without its 3'UTR could prevent the miR-28-5p from inducing the inhibition of glioma tumor cells. CONCLUSIONS: Our findings showed that miR-28-5p could suppress the growth, invasion and migration of glioma cells by suppressing the SphK1 expression. The results demonstrated that miR-28-5p might serve as an important potential therapeutic target for glioma.

Androcam, a Drosophila calmodulin-related protein, is expressed specifically in the testis and decorates loop kl-3 of the Y chromosome.

The Drosophila genome encodes a protein that is 68% identical to Drosophila calmodulin (Cam). We show here that this Cam-related gene is specifically expressed in the germ-line of the testis, leading to the name Androcam (Acam). Early in spermatogenesis Acam accumulates on one of the chromatin loops of the Y chromosome, kl-3. This association with kl-3 may indicate an RNA processing-related role for Acam and/or could reflect an unusual storage/assembly function hypothesized for the Y loops. After meiosis Acam is detectable in developing sperm tail cytoplasm, where at least some of the protein is not tightly associated with tubulin. Late in spermiogenesis, some Acam staining overlaps the periphery of the investment cones, actin-containing structures hypothesized to support the motor function for cytoplasmic stripping of the tail. Acam cannot be detected in mature sperm by immunolocalization, but immunoblotting established that Acam is present in sperm stored in mated females, suggesting epitope masking during final maturation. Proteins more related to Acam than Cam are present in the testes of other Drosophila species and a mammalian species, the mouse.

Conformational and metal-binding properties of androcam, a testis-specific, calmodulin-related protein from Drosophila.

Androcam is a testis-specific protein of Drosophila melanogaster, with 67% sequence identity to calmodulin and four potential EF-hand calcium-binding sites. Spectroscopic monitoring of the thermal unfolding of recombinant calcium-free androcam shows a biphasic process characteristic of a two-domain protein, with the apo-N-domain less stable than the apo-C-domain. The two EF hands of the C-domain of androcam bind calcium cooperatively with 40-fold higher average affinity than the corresponding calmodulin sites. Magnesium competes with calcium binding [Ka(Mg) approximately 3 x 10(3) M(-1)]. Weak calcium binding is also detected at one or more N-domain sites. Compared to apo-calmodulin, apo-androcam has a smaller conformational response to calcium and a lower alpha-helical content over a range of experimental conditions. Unlike calmodulin, a tryptic cleavage site in the N-domain of apo-androcam remains trypsin sensitive in the presence of calcium, suggesting an altered calcium-dependent conformational change in this domain. The affinity of model target peptides for androcam is 10(3)-10(5) times lower than for calmodulin, and interaction of the N-domain of androcam with these peptides is significantly reduced. Thus, androcam shows calcium-induced conformational responses typical of a calcium sensor, but its properties indicate calcium sensitivity and target interactions significantly different from those of calmodulin. From the sequence differences and the altered calcium-binding properties it is likely that androcam differs from calmodulin in the conformation of residues in the second calcium-binding loop. Molecular modeling supports the deduction that there are significant conformational differences in the N-domain of androcam compared to calmodulin, and that these could affect the surface, conferring a different specificity on androcam in target interactions related to testis-specific calcium signaling functions.

Calcium-binding proteins and development.

The known roles for calcium-binding proteins in developmental signaling pathways are reviewed. Current information on the calcium-binding characteristics of three classes of cell-surface developmental signaling proteins (EGF-domain proteins, cadherins and integrins) is presented together with an overview of the intracellular pathways downstream of these surface receptors. The developmental roles delineated to date for the universal intracellular calcium sensor, calmodulin, and its targets, and for calcium-binding regulators of the cytoskeleton are also reviewed.