Implementation of an Ultrasensitive Biomolecular Controller for Enzymatic Reaction Processes With Delay Using DNA Strand Displacement.

In this article, a set of abstract chemical reactions has been employed to construct a novel nonlinear biomolecular controller, i.e, the Brink controller (BC) with direct positive autoregulation (DPAR) (namely BC-DPAR controller). In comparison to dual rail representation-based controllers such as the quasi sliding mode (QSM) controller, the BC-DPAR controller directly reduces the number of CRNs required for realizing an ultrasensitive input-output response because it does not involve the subtraction module, reducing the complexity of DNA implementations. Then, the action mechanism and steady-state condition constraints of two nonlinear controllers, BC-DPAR controller and QSM controller, are investigated further. Considering the mapping relationship between CRNs and DNA implementation, a CRNs-based enzymatic reaction process with delay is constructed, and a DNA strand displacement (DSD) scheme representing time delay is proposed. The BC-DPAR controller, when compared to the QSM controller, can reduce the number of abstract chemical reactions and DSD reactions required by 33.3% and 31.8%, respectively. Finally, an enzymatic reaction scheme with BC-DPAR controller is designed using DSD reactions. According to the findings, the enzymatic reaction process's output substance can approach the target level at a quasi-steady state in both delay-free and non-zero delay conditions, but the target level can only be achieved during a finite-time period, mainly due to the fuel stand depletion.

Chromosome genome assembly and annotation of the yellowbelly pufferfish with PacBio and Hi-C sequencing data.

Pufferfish are ideal models for vertebrate chromosome evolution studies. The yellowbelly pufferfish, Takifugu flavidus, is an important marine fish species in the aquaculture industry and ecology of East Asia. The chromosome assembly of the species could facilitate the study of chromosome evolution and functional gene mapping. To this end, 44, 27 and 50 Gb reads were generated for genome assembly using Illumina, PacBio and Hi-C sequencing technologies, respectively. More than 13 Gb full-length transcripts were sequenced on the PacBio platform. A 366 Mb genome was obtained with the contig of 4.4 Mb and scaffold N50 length of 15.7 Mb. 266 contigs were reliably assembled into 22 chromosomes, representing 95.9% of the total genome. A total of 29,416 protein-coding genes were predicted and 28,071 genes were functionally annotated. More than 97.7% of the BUSCO genes were successfully detected in the genome. The genome resource in this work will be used for the conservation and population genetics of the yellowbelly pufferfish, as well as in vertebrate chromosome evolution studies.

Ouabain targets the Na+/K+-ATPase α3 isoform to inhibit cancer cell proliferation and induce apoptosis.

Ouabain has been used for the treatment of heart failure and atrial fibrillation. Its potential anticancer effect has also attracted great interest. The aim of the present study was to evaluate the anticancer effect of ouabain and investigate its molecular target. The effects of ouabain on the viability of and induction of cellular death on OS-RC-2 renal cancer cells were examined using the MTT assay and acridine orange/ethidium bromide staining. The levels of Ca2+ and reactive oxygen species were determined using Fura-3-acetoxymethyl ester and dichloro-dihydro-fluorescein diacetate probes, respectively. Apoptosis was examined using annexin V-fluorescein isothiocyanate/propidium iodide staining and western blotting. The expression profile of the different Na+/K+-ATPase (NKA) isoforms in NCI-H446 small cell lung cancer cells was determined using immunocytochemistry and reverse transcription polymerase chain reaction analysis. In the present study, it was demonstrated that ouabain inhibited cancer cell proliferation and induced apoptosis while no significant difference in the expression of NKA α1 and α3 isoforms was detected following 48 h of ouabain treatment. Furthermore, expression of NKA α3 but not the α1 isoform was associated with ouabain sensitivity. The results of the present study indicated that ouabain targets the NKA α3 isoform, inhibits cancer cell proliferation and induces apoptosis.

Defective insulin signaling and the protective effects of dimethyldiguanide during follicular development in the ovaries of polycystic ovary syndrome.

It is established that the physiological effects of insulin are primarily mediated by the insulin signaling pathway. However, a defective insulin signaling is closely associated with the clinical manifestations of polycystic ovary syndrome (PCOS), which include excess androgen levels, insulin resistance and anovulation, and is involved in the pathophysiology of PCOS at the molecular level. Dimethyldiguanide (DMBG) has been widely employed to alleviate reproduction dysfunction in women with PCOS, however, the exact mechanism of this effect remains unclear. The objective of the present study was to investigate the effects of DMBG on the expression of the insulin signaling pathway in the ovaries of rats with PCOS, and to identify the potential underlying molecular mechanisms of these effects in PCOS. In the present study, a PCOS rat model was induced by letrozole, and successful establishment of the model was confirmed by examining ovarian histology and determining serum testosterone levels, by hematoxylin and eosin staining and ELISA, respectively. Subsequently, the expression of two key elements of insulin signaling, insulin receptor substrate (IRS)­2 and phosphatidylinositol 3­kinase (PI3K), was determined by immunohistochemistry and western blot analysis. The results demonstrated that IRS­2 and PI3K expression was markedly decreased in PCOS ovaries, which was rescued by DMBG treatment. These results indicate that IRS­2/PI3K signaling may be involved in the development of PCOS and the therapeutic effects of DMBG on PCOS. To further confirm the effects of DMBG on insulin signaling expression during this process, the expression of an additional two downstream proteins, phosphoinositide­dependent kinase­1 (PDK­1) and the mammalian target of rapamycin (mTOR), was also investigated in the present study, and the results demonstrated that the expression of PDK­1 and mTOR was significantly reduced in PCOS ovaries and increased following DMBG treatment, further indicating that altered insulin signaling may have an important role in the development and treatment of PCOS. In conclusion, the results of the present study indicate that the reduced expression of proteins involved in insulin signaling may contribute to the development of the clinical features of PCOS, and DMBG reverses reduced expression of insulin signaling components, by a mechanism that is yet to be determined, to attenuate certain symptoms of PCOS, such as obesity. To the best of our knowledge, the present study is the first to provide data regarding the detailed changes of insulin signaling during the development and treatment of PCOS, and may provide an important reference for clinical PCOS treatment.

Digitoxin synergizes with sorafenib to inhibit hepatocelluar carcinoma cell growth without inhibiting cell migration.

Sorafenib is a chemotherapeutic agent approved for the treatment of hepatocellular carcinoma (HCC) in China. Digitoxin is a cardiotonic drug, which has been demonstrated to exhibit anticancer effects in a number of cancers, but not in HCC. The aim of the present study was to evaluate the combinational effect of sorafenib and digitoxin on the treatment of HCC and to investigate the relevant molecular mechanisms of action that underlie these effects. The proliferation, cell death and migration of HCC cell lines, HepG2 and BEL­7402, were examined using MTT, acridine orange/ethidium bromide staining and scratch wound healing assays, respectively. In addition, alterations in the expression of phosphorylated-extracellular signal-regulated kinase (ERK), hypoxia­inducible factor 1­α (HIF­1α), hypoxia­inducible factor 2­α (HIF­2α) and vascular endothelial growth factor (VEGF) were measured prior to and following drug application using western blot analysis. Digitoxin and sorafenib synergistically inhibited cell viability, but did not inhibit migration, which was potentially mediated by suppression of ERK and hypoxia signaling. In downstream signaling pathways, the activity of ERK was synergistically suppressed by combinatorial treatment of HepG2 and BEL­7402 cells with sorafenib and digitoxin. In addition, the expression of HIF­1α, HIF­2α and VEGF was synergistically downregulated by combinational treatment.

3D assessment of lip scarring and residual dysmorphology following surgical repair of cleft lip and palate: a preliminary study.

OBJECTIVE: To evaluate lip scarring and the three-dimensional (3D) lip morphology following primary reconstruction in children with unilateral cleft lip and palate (UCLP) relative to contemporaneous noncleft data. DESIGN: Retrospective, cross-sectional, controlled study. SETTING: Glasgow Dental Hospital and School, University of Glasgow, U.K. PATIENTS AND PARTICIPANTS: Three groups of 10-year-old children: 51 with UCLP, 43 UCL (unilateral cleft lip), and 68 controls. METHODS: Three-dimensional images of the face were recorded using stereo cameras on a two-pod capture station, and 3D coordinates of anthropometric landmarks were extracted from the facial images. A novel method was applied to quantify residual scarring and the associated lip dysmorphologies. The relationships among outcome measures were investigated. RESULTS: Residual lip dysmorphologies were more pronounced in UCLP cases. The width of the Cupid's bow was increased due to lateral displacement of the christa philteri left (cphL) in both UCL and UCLP patients. In the upper part of the lip, the nostril base was significantly wider in UCLP cases when compared with UCL cases and controls. Scar redness was more pronounced in UCL than in UCLP cases. No relationship could be identified between lip scarring and other measurements of lip dysmorphology. CONCLUSIONS: Stereophotogrammetry, together with associated image analysis, allow early detection of residual dysmorphology following cleft repair.

[Polysaccharides from Scurrula parasitica L. inhibit sarcoma S180 growth in mice].

To study the anti-tumor activity of Scurrula parasitica polysaccharides (SP). Water extraction and ethanol precipitation were used to isolate SP from S. parasitica leaf. S180, K562 and HL-60 cell lines proliferation inhibition by SP were detected by MTT assay. The expressions of Ki-67, Cyclin D1, Bax and Bcl-2 protein in the sarcoma S180 tissues were detected by immunohistochemistry technique to approach the anti-tumor mechanism of SP+ SP could not inhibit cancer cell proliferation. SP ip could inhibit the growth of sarcoma S180 in mice, 100 mg x kg(-1) x d(-1). SP ip was the optimal dose on inhibiting S180 growth, with the tumor inhibition rate of 54%. The expression of Ki-67, Cyclin D1, Bax and Bcl-2 protein in the sarcoma S180 tissues were detected by immunohistochemistry technique to approach the anti-tumor mechanism of SP. The result showed that SP could down-regulate the expression of Ki-67, CyclinD1 and Bcl-2 protein, and up-regulate the expression of Bax protein. It indicted that inhibiting cancer cell proliferation and promoting cancer cell apoptosis in vivo maybe one of the anti-cancer mechanisms of SP.

[Selective effect of nispex in inhibiting human cancer cell proliferation and inducing cell apoptosis].

OBJECTIVE: To observe the in vitro anticancer effect of Nispex, the total flavonoids extract from Scurrula parasitic L. METHODS: The cell proliferation inhibitory effects of Nispex on various kinds of tumor cells or non-tumor cells in human and rats were detected with MTT assay and colony forming assay respectively, the cell apoptosis induced by Nispex was detected by AO/EB fluorescence staining, TUNEL assay and AnnexinV-FITC/PI flow cytometry. RESULTS: Nispex could significantly inhibit human cancer cell proliferation and induce human cancer cell apoptosis, especially to the proliferative cell group, but its inhibition on human non-tumor cell was insignificant, and showed no effect on murine cancer cells in the tested scope. CONCLUSION: Nispex is a nature plant extract which shows good selectivity for killing human cancer cell.

[Study on cytotoxic activities on human leukemia cell line HL-60 by flavonoids extracts of Scurrula parasitica from four different host trees].

OBJECTIVE: To compare the anticancer effects of flavonoids extracts of Scurrula parasitica from different host trees in vitro. METHOD: 80% ethanol extracts of S. parasitica parasitizing on Nernium indicum, Morus alba, Opsmanthus fragrans, and Sapindus mulorossi were purified by polyamides column chromatography, and the eluates of 30%, 50%, 70% and 90% ethanol were mixed as flavonoids extracts. Human acute myeloid leukemia cell line HL-60 was used to evaluate the cytotoxicity induced by flavonoids extracts of S. parasitica L with MTT assay. Apoptosis was detected by AO/EB fluorescence staining and DNA fragmentation analysis, apoptosis rates and cell cycle distribution were detected by flow cytometry analysis. RESULT: Extract of S. parasitica parasitizing on N. indicum (NISPEX) was the most sensitive to HL-60 cells of the 4 different host trees, the IC50 value being 0.60 mg x L(-1); and extract of S. parasitica parasitizing on M. alba took the second place, the IC50 value, being 2.49 mg x L(-1); extract of S. parasitica parasitizing on O. fragrans had no effectiveness as high as 50 mg x L(-1) concentration. NISPEX induced HL-60 cell apoptosis and inhibited the cell proliferation in dose and time-dependent manner. Cell cycles were arrested at G0-G1 phase after treated with NISPEX. CONCLUSION: Anticancer effects of S. parasitica correlated with the host trees. Flavonoids extracts of S. parasitica parasitizing on N. indicum exhibited comparatively better anticancer activity in vitro among the host trees studied. NISPEX is found to be a good candidate for anticancer.

A functional-based segmentation of human body scans in arbitrary postures.

This paper presents a general framework that aims to address the task of segmenting three-dimensional (3-D) scan data representing the human form into subsets which correspond to functional human body parts. Such a task is challenging due to the articulated and deformable nature of the human body. A salient feature of this framework is that it is able to cope with various body postures and is in addition robust to noise, holes, irregular sampling and rigid transformations. Although whole human body scanners are now capable of routinely capturing the shape of the whole body in machine readable format, they have not yet realized their potential to provide automatic extraction of key body measurements. Automated production of anthropometric databases is a prerequisite to satisfying the needs of certain industrial sectors (e.g., the clothing industry). This implies that in order to extract specific measurements of interest, whole body 3-D scan data must be segmented by machine into subsets corresponding to functional human body parts. However, previously reported attempts at automating the segmentation process suffer from various limitations, such as being restricted to a standard specific posture and being vulnerable to scan data artifacts. Our human body segmentation algorithm advances the state of the art to overcome the above limitations and we present experimental results obtained using both real and synthetic data that confirm the validity, effectiveness, and robustness of our approach.

[First-line chemotherapy of weekly paclitaxel/cisplatin for un-resectable non-small cell lung cancer].

BACKGROUND: Chemotherapy is an important treatment for un-resectable lung cancer patients. The aim of this study is to investigate effects and safety of weekly paclitaxel/cisplatin as first-line chemotherapy in un-resectable non-small cell lung cancer (NSCLC). METHODS: Thirty-eight initially treated patients (male/female: 20/18) with un-resectable NSCLC were enrolled for the study. They were at ages ranging from 33 to 82 years old with ECOG PS of 0 to 2. Paclitaxel 80mg/m² was given by intravenous infusion on 1st and 8th day, and cisplatin 25mg/m² on 2th to 6th days, 3 to 4 weeks was one cycle. The responses and toxicity of chemotherapy were evaluated after six cycles and the patients were followed up. RESULTS: In 38 patients, partial response and complete response were observed in 21 cases and 1 case, respectively with overall response rate of 57.9%. The response rate in cases with ECOG of 0 to 1 was significantly higher than those with ECOG PS of 2 (69.0% vs 22.2%). Median survival time was 14 months and 1-, 2- and 3-year survival rate were 63.8%, 29.5% and 16.2% respectively. Main Toxicities were leucopenia and alopecia, and all patients could tolerate the side effects and there was no drug-related death associated with myelotoxicity. CONCLUSIONS: Regimen of paclitaxel/cisplatin was efficient and safe as the first-line treatment for un-resectable NSCLC patients.