Messenger ribonucleic acids (mRNAs) comprise a class of small nucleic acids carrying genetic information, which exhibit very important role in medical research and diagnosis. If only the mean mRNA expression levels of the mRNA population are considered in medical research, important information linking mRNA expression and cellular function may be lost. Single-cell analysis provides valuable insights into studying its heterogeneity, signaling, and stochastic gene expression. In this study, a \"bunge bedstraw herb\"-type DNA machine based on DNAzyme catalyzing coupled clamping hybrid chain reaction (c-HCR) is presented. In the DNA machine, a bunge bedstraw herb-type DNA structure was first formed by hybridizing a core junction scaffold cruciform probe to a hairpin probe that can trigger the c-HCR via a target molecule in four directions. This approach can reduce the detection limit of mRNA to 5 × 10-15 M. Absolute quantification of survivin mRNA in individual cells was achieved using the DNA machine on a microfluidic chip electrophoresis platform. The reported method represents an unprecedented single-cell analysis platform for single-cell biology studies.

To accurately evaluate expression levels of target genes, stable internal reference genes is required for normalization of quantitative real-time PCR (qRT-PCR) data. However, there have been no systematical investigation on the stability of reference genes used in the bedstraw weed, Galium aparine L. (BGA). In this study, the expression profiles of seven traditionally used reference genes, namely 18S, 28S, ACT, GAPDH, EF1α, RPL7 and TBP in BGA were assessed under both biotic (developmental time and tissue), and abiotic (temperature, regions and herbicide) conditions. Four analytical algorithms (geNorm, Normfinder, BestKeeper and the ΔCt method) were used to analyze the suitability of these genes as internal reference genes. RefFinder, a comprehensive analytical software, was used to rank the overall stability of the candidate genes. The optimal normalization internal control genes were ranked as: 28S and RPL7 were best for all the different experimental conditions (developmental stages, tissues, temperature, regions and herbicide treatment); 28S and RPL7 for developmental stages; TBP and GAPDH for different tissues; 28S and GAPDH were relatively stable for different temperature; 28S and TBP were suitable for herbicide treatment. A specific set of reference genes were recommended for each experimental condition in BGA.

Galium L. is the largest genus in the tribe Rubieae, with about 667 species distributed worldwide. Previous researches mainly focused on species from the Americas and Europe. In the present paper, we greatly increased the number of samples examined from eastern Asia (especially China), representing the most comprehensive sampling of Galium to date. A total of 194 species and variations (subspecies) of Galium were sampled to determine phylogenetic relationships, using two nuclear and five chloroplast markers. Our data are largely consistent with all previous phylogenetic results and confirmed that Galium is non-monophyletic, as are most of its sections. Most members of Galium, including the Chinese taxa, fall into three large clades mixed with other genera from the Galium s.l. group; the exception being the distinct Galium paradoxum Maxim., the first diverged lineage in the Galium s.l. group, which was treated as a new genus (Pseudogalium L.-E. Yang, Z.-L. Nie & H. Sun, gen. nov.). The Galium s.s is a well-supported clade comprised entirely of Galium species, usually with six or more leaves per whorl, mostly from the Old World. Samples from G. maximowiczii (Kom.) Pobed, G. sect. Depauperata and sect. Aparinoides, together with a few from Asperula sect. Glabella and Microphysa (Schrenk ex Fisch. & C.A. Mey.) Pobed., form the second clade. The third clade comprises taxa purely from Galium that usually have four leaves per whorl, from both the New and Old World. Our results also indicated that the monotypic genus Microphysa should be retained and clarified phylogenetic relationships of some specific confused taxa from China. Unlike prior inferences, the combination of opposite leaves associated with two stipules is proposed as the ancestral characteristic of the Galium s.l. group and even the tribe. In addition, the shapes of different corolla and inflorescence types are important for distinguishing some taxa within Rubieae.

Intercropping affects heavy-metal uptake in plants. In this study, three cadmium (Cd)-accumulator species (Stellaria media, Malachium aquaticum, and Galium aparine) were intercropped together in Cd-contaminated soil to study the effects of intercropping on co-remediation. Mutual intercropping significantly increased the biomasses of S. media, M. aquaticum, and G. aparine compared with their respective monocultures. The photosynthetic pigment contents of three species were not significantly affected by mutual intercropping. Mutual intercropping did not increase the Cd contents in roots and shoots of G. aparine, but it increased the Cd contents in roots of S. media and M. aquaticum. It also decreased the Cd contents in shoots of S. media and M. aquaticum. Only G. aparine intercropped with M. aquaticum and three-species intercropping increased Cd-accumulation levels in whole plants of each species. Only S. media intercropped with M. aquaticum and three-species intercropping increased the Cd accumulation at the whole plant level in every plant in a single pot, with S. media intercropped with M. aquaticum showing the greatest increase. Therefore, S. media intercropped with M. aquaticum and three-species intercropping may improve the efficiency of phytoremediation of Cd-contaminated soil, with S. media intercropped with M. aquaticum representing the best combination.

OBJECTIVE: To investigate the effects of flavone from Galium verum L (FGVL) on hydrogen peroxide induced oxidative injury in human umbilical vein endothelial cells (HUVEC), and explore related mechanisms.
METHODS: HUVEC were divided into five groups: control group (1640 complete medium), injured group (HUVEC treated with 100 μmol/L hydrogen peroxide for 4 h), FGVL group (HUVEC treated with 12.5 mg/L FGVL (group F1), 25.0 mg/L (group F2), 50.0 mg/L (group F3) for 24 h before hydrogen peroxide). The nitric oxide content was measured by nitric acid reductase method. The 6-keto-Prostacyclin-F1α (6-keto-PGF1α), thromboxane B2 (TXB2), interleukin(IL)-6 and IL-22 were determined by ELISA.mRNA expression of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and endothelial nitric oxide synthase (eNOS) was detected by RT-PCR.Protein expression of p-Akt (ser(473)) and p-eNOS (ser(1177)) was determined by Western blot.Cell apoptosis was observed with fluorescence microscope after Hoechst33258 staining.
RESULTS: (1) The contents of nitric oxide were significantly lower in the injured group than in the control group ((34.11±1.78) μmol/L vs. (74.81±2.93) μmol/L, P<0.05), which was significantly increased in group F2 ((41.86±2.32) μmol/L) and group F3 ((62.79±1.16) μmol/L) compared with injured group (both P<0.05). (2)The secretion level of 6-keto-PGF1α was significantly lower in the injured group ((44.84±3.87) ng/L) than in the control group ((82.38±3.98) ng/L, P<0.05), which was significantly increased in group F1 ((52.76±1.78) ng/L), FGVL 2 group which was(56.58±1.44) ng/L and FGVL 3 group which was(67.78±2.02) ng/L than that of injured group(all P<0.05). The secretion level of TXB2 was significantly higher in the injured group((43.37±3.96) ng/L) than in the control group ((25.56±1.75) ng/L, P<0.05), which was significantly reduced group F2 group ((32.41±1.68) ng/L) and group F3 ((28.23±2.15) ng/L) than that of injured group(both P<0.05). (3) The contents of IL-6 and IL-22 were significantly higher in the injured group ((539.74±11.63) ng/L) and ((23.70±3.05) ng/L, respectively) than in the control group ((288.67±19.52) ng/L) and ((23.70±3.05) ng/L, respectively, both P<0.05). The contents of IL-6 were significantly lower in group F1, F2 and F3 compared to that of injured group(all P<0.05). The contents of IL-22 were significantly lower in group F2 and F3 than that of injured group(both P<0.05). (4)The relative levels of PI3K mRNA and eNOS mRNA in injured group (0.68±0.09 and 0.22±0.03, respectively) were significantly lower compared to control group(0.81±0.12 and 0.63±0.11, respectively, both P<0.05), PI3K mRNA in group F2 (0.76±0.03) and group F3 (PI3K mRNA 0.83±0.06) as well as eNOS mRNA in group F1 (0.37±0.08), F2 (0.53±0.04) and F3 (0.56±0.09) than those of injured group(all P<0.05). The mRNA expression of Akt was similar among groups (P>0.05). (5) The relative levels of p-Akt (ser(473)) and p-eNOS (ser(1177)) in injured group (0.48±0.05 and 0.23±0.03, respectively) were significantly lower compared to control group (0.71±0.12 and 0.66±0.05, respectively, both P<0.05), which was up-regulated in group F1, F2 and F3 groups compared to injured group(all P<0.05). (6) The cell apoptosis rate in injured groups was significantly higher compared to control group which ((63.67±11.37)% vs. (4.67±1.15)%, P<0.05) which was significantly reduced in group F1((43.33±4.16)%), F2((18.33±4.93)%) and F3((15.67±2.08)%) compared to injured group(all P<0.05).
CONCLUSIONS: The FGVL can reduce hydrogen peroxide induced oxidative injury in HUVEC by increasing the level of nitric oxide through PI3K/Akt/eNOS pathway.

Portulaca oleracea L. has been used as folk medicine in different countries to treat different ailments in humans. P. oleracea L. polysaccharide (POL-P), extracted from P. oleracea L., is found to have bioactivities such as hypoglycemic and hypolipidemic activities, antioxidant and antitumor activities. In our study, a water-soluble polysaccharide (POL-P3b) was successfully purified from Galium verum L. by DEAE cellulose and Sephadex G-200 column chromatography. To evaluate the anticancer efficacy and associated mechanisms of POL-P3b on cervical cancer in vitro and in vivo, we showed that treatment of HeLa cell with POL-P3b inhibited cell proliferation. In addition, POL-P3b significantly inhibited tumor growth in U14-bearing mice. Further analysis indicated that POL-P3b possesses the activity of inhibiting cervical cancer cell growth in vitro and in vivo at a concentration- and time-dependent manner, and the mechanisms were associated with Sub-G1 phase cell cycle arrest, triggering DNA damage and inducing apoptosis.

GaN-functionalized rare earth (Eu3+ and Tb3+) organic/inorganic mesoporous nanocomposites have been successfully synthesized (designated as RE(L-SBA-15)3(L-GaN); RE=Eu, Tb; L=TAA-Si, BTA-Si). The organosilane precursor materials (L-SBA-15) are synthesized by co-condensation of tetraethylorthosilicate (TEOS) and the functionalized β-diketones (TAA-Si and BTA-Si) in the presence of Pluronic P123 surfactant as a template. The modified β-diketones ligands are also used to covalently bond with surface-modified GaN and formed another precursor L-GaN. Both of the precursors can coordinate with rare earth ions to synthesize the final mesoporous materials via a sol-gel process. FTIR, TEM, XRD, and nitrogen (N2) adsorption/desorption measurements are employed to characterize the mesostructure of RE(L-SBA-15)3(L-GaN). The luminescence properties and thermogravimetric analysis of all the prepared materials are characterized in detail, and the results reveal that a series of uniformed mesopore structure hybrid materials has been achieved. The mesoporous material Eu(BTA-Si-SBA-15)3(BTA-Si-GaN) has better luminescence intensity, higher quantum efficiency, and longer lifetime than Eu(TAA-Si-SBA-15)3(TAA-Si-GaN). While the nanocomposite Tb(TAA-Si-SBA-15)3(TAA-Si-GaN) revealed the strongest characteristic emission of Tb ions than Tb(BTASiSBA-15)(BTASiGaN), the excellent luminescent properties and thermal stability enable the hybrid mesoporous material to have potential applications in optical field.

Hook-like surface structures, observed in some plant species, play an important role in the process of plant growth and seed dispersal. In this study, we developed an elastic model and further used it to investigate the mechanical behaviour of fruit hooks in four plant species, previously measured in an experimental study. Based on Euler-Bernoulli beam theory, the force-displacement relationship is derived, and its Young\'s modulus is obtained. The result agrees well with the experimental data. The model aids in understanding the mechanics of hooks, and could be used in the development of new bioinspired Velcro-like materials.

Diosmetin (DGVL) extracted from the traditional Chinese herb Galium verum L. has been found to have anticancer activity. In this study, the effects of DGVL on the thymus of U14-bearing mice were investigated. Using flow cytometry, peripheral blood lymphocytes were characterized based on the expression of surface markers for T helper cells (CD4(+)) and T suppressor cells (CD8(+)). Serum levels of tumor necrosis factor α (TNF-α), interleukin-2 (IL-2), IL-10, and transforming growth factor β1 (TGF-β1) and a cell proliferation assay were determined with an enzyme-linked immunosorbent assay. The expression of Fas and Fas ligand (FasL) on the thymus was determined by Western blotting. Our results showed that DGVL inhibited tumor growth and significantly increased the thymus weight compared with the control. Also, DGVL elevated serum levels of IL-2 and significantly reduced levels of TNF-α, TGF-β1, and IL-10 in a dose-dependent manner. Histological study and terminal dUTP nick end labeling staining results showed that DGVL protected thymus tissue against the onslaught of tumor growth by inhibiting thymus lymphocyte apoptosis. The cell proliferation assay revealed that DGVL might promote more thymus lymphocytes towards proliferation. Furthermore, the ratio of CD4(+)/CD8(+) T lymphocytes was significantly increased from 0.69 to 2.29 by treatment with DGVL. Immunoblotting analyses revealed that the expression of Fas and FasL on the thymus was lower in mice in the DGVL treatment group than in the control mice. In conclusion, DGVL can inhibit tumor growth and protect tumor-induced apoptosis of the thymus, and the mechanism is closely associated with reduced cell death in the thymus and a Fas-FasL-dependent pathway.

OBJECTIVE: To study the influence of C-terminal domain of ADAMTS13 on its cleaving activity.
METHODS: The full-length wild-type (WT) and C-terminal domain truncated type (TT, TSP8 + CUB domains were deleted) of human ADAMTS13 recombinant protein were transfected into and permanent expressed on Hela cells. Western blot and R-CBA were used to directly detect the activities of the two recombinant proteins under the static and stressed condition respectively. ELISA was used to compare the binding abilities of the two proteins by coating with vWF.
RESULTS: The recombinant proteins were identified by Western blot with anti-his-tag or anti-ADAMTS13 antibodies. With pretreatment of 1.5 M urea, the enzyme activity of TT was significantly higher than that of WT, and so did in binding ability with vWF While, only WT could cleave vWF under high stress.
CONCLUSIONS: The distal carboxyl-terminal TSP8 together with CUB domains of ADAMTS13 may affect the enzyme activity by regulating the binding of ADAMTS13 to vWF in different conditions, and they are very important for the enzyme activity under high stress force condition.