Notoginsenoside R1 induces DNA damage via PHF6 protein to inhibit cervical carcinoma cell proliferation.

Notoginsenoside R1 (NGR1), a monomer of Traditional Chinese medicine, is from the Panax notoginsenoside complex, and has been reported to inhibit the proliferation of various types of cancer. However the mechanism underlying NGR1­mediated inhibition of cervical carcinoma cell proliferation remains unclear. Therefore, the current study aimed to investigate the antitumor effects of NGR1 on cervical carcinoma cell lines (CaSki and HeLa cells) in vitro. The Cell Counting Kit­8 and soft agar cell colony formation assay results revealed that NGR1 suppressed the viability and the number colonies of CaSki and HeLa cells, respectively. Furthermore, the DAPI staining, flow cytometry and western blotting results revealed that NGR1 induced cervical carcinoma cell apoptosis, cell cycle arrest in the S phase, upregulation of cyclin A2 and CDK2 expression levels, and downregulation of cyclin D1 expression levels. To further investigate the mechanisms of NGR1, DNA­damage­related proteins, including H2A.X variant histone (H2AX), ATR serine/threonine kinase (ATR) and p53, and the nucleolus protein, plant homeodomain finger protein 6 (PHF6) were analyzed. The results indicated that NGR1 triggered the phosphorylation of H2AX and ATR in a dose­ and time­dependent manner, and downregulated the expression level of PHF6 and upregulated the expression level of p53 in a dose­ and time­dependent manner. In conclusion, the findings of the present indicated that NGR1 may inhibit the viability of cervical carcinoma cells and induce cell apoptosis via DNA damage, which may be activated by the downregulation of PHF6 expression levels, and the subsequent triggering of the phosphorylation of H2AX and ATR. In addition, NGR1 may exert an ability to arrest cervical carcinoma cells in the S phase and upregulate the expression levels of cyclin A2 and CDK2. Therefore, NGR1 may serve as a novel chemotherapeutic agent for cervical carcinoma.

Determination of copper ions in herbal medicine based on click chemistry using an electronic balance as a readout.

The amount of copper affects the quality of herbal medicine greatly, it is necessary to develop some simple and sensitive methods to detect copper for the remote or resource-limited area. An electronic balance is one of the most familiar equipment that can be found nearly in all laboratories. The presence of Cu(i) can catalyze azide-alkyne cycloaddition reaction (called as click chemistry) with high efficiency. In this study, a simple method had been developed to detect copper ions in herbal medicine using an electronic balance as a readout device based on click chemistry. Cu(ii) is reduced to Cu(i) by sodium ascorbate in situ, which induces the "click" reaction between azido-DNA modified magnetic beads (MB-DNA) and alkynyl-DNA modified platinum nanoparticles (Pt NP-DNA) and results in the fixing of the platinum nanoparticles on the beads (called as MB-Pt NPs). MB-Pt NPs can be separated by a magnetic frame easily and transferred into a drainage reaction device containing hydrogen peroxide. Then, hydrogen peroxide can be decomposed by Pt NPs modified on MB to generate oxygen, which increases the pressure in the drainage reaction device and forces the water in the system to be discharged. The weight of the discharged water can be easily and accurately measured by an electronic balance. The weight of the water has a linear relationship with Cu(ii) in the range of 2.0-200 µM and a detection limit of 0.83 µM under 30 min of collected time. This method does not need complicated and expensive instruments, skilled technicians, and a complex data processing process. The proposed method had been applied to detect copper ions in herbal medicine with satisfactory results.

Rapid authentication of Pseudostellaria heterophylla (Taizishen) from different regions by near-infrared spectroscopy combined with chemometric methods.

Pseudostellaria heterophylla is a very popular traditional Chinese medicine herb, also called "Taizishen." Discrimination of P. heterophylla from different regions is critical for ensuring the effectiveness of drug use, because the drug effects of P. heterophylla from different regions are diversity of each other. To discriminate P. heterophylla from different regions rapidly and effectively, a model extracted by competitive adaptive reweighted sampling (CARS) was established. Original spectra of P. heterophylla in wave number range of 10,000 to 4,000 cm-1 were acquired. Orthogonal partial least squares discriminant analysis (OPLS-DA) was also used to establish a suitable model. CARS was performed for extracting key wave number variables. We found that the near-infrared spectrum of a series of samples analyzed by Row-center-SG, CARS, and OPLS-DA can effectively distinguish the P. heterophylla from different regions, and the accuracy of OPLS-DA model is also satisfactory in terms of good discrimination rate. These results show that the Row-center-SG, CARS, and OPLS-DA model can be used to identify the P. heterophylla from different regions. PRACTICAL APPLICATION: According to our research results, we can draw a conclusion that our research results may be used to distinguish the traditional Chinese medicine from those from different places of origin and the powder with similar appearance.

On-spot surface enhanced Raman scattering detection of Aflatoxin B1 in peanut extracts using gold nanobipyramids evenly trapped into the AAO nanoholes.

Simple, rapid, convenient, and economical surface enhanced Raman scattering (SERS) substrate is developed for on-site evaluation of Aflatoxin B1 (AFB1) in food matrix using handheld Raman Spectrometer. Self-assembly of gold nanobipyramids (Au NBPs) into the nanoholes of anodic aluminum oxide (AAO) template/pattern using 'drop-dry' approach provides a reliable pathway for the rapid fabrication of highly active and uniform SERS substrate. It shows enhanced and reproducible SERS signals towards the probe molecule, 4-aminothiophenol (4-ATP) with a relative standard deviation (RSD) of less than 10% and an average enhancement factor (EF) of 1 × 108. For practical application, the proposed method is demonstrated for the detection of aflatoxin B1 (AFB1) in peanut extracts. The results show that the AFB1 in peanut extracts can be identified within 1 min, with a limit of detection of 0.5 µg/L. Compared with conventional ELISA based AFB1 analysis, our method is much more efficient (1 min versus >30 min).

Antibacterial mechanism of Tetrastigma hemsleyanum Diels et Gilg's polysaccharides by metabolomics based on HPLC/MS.

Tetrastigma hemsleyanum Diels et Gilg (THDG) is used as a Chinese traditional anti-inflammatory medicine for about thousands of years. In this work, Tetrastigma hemsleyanum Diels et Gilg's polysaccharide (TP) can inhibit E. coli's growth in initial dosing period. Compared with the antibacterial effect of Achyranthe's polysaccharide (AP) from their metabolic profile, it's obviously that their metabolic sites for E. coli were inconsistent. Moreover, TP could not only increase the level of fructose-6-phosphate (F6P), decrease the level of fructose-1,6-diphosphate (FBP), but also charge the amount of the two differential metabolic with the change of the concentration and the dosing time. Actually, F6P could transform into FBP by catalyze of 6-phosphofructokinase-1(6-PFK-1), which is an important process in glycolysis. Furthermore, FBP was considered have positively correlated with E. coli's growth rate. Therefore, TP can inhibit the E. coli's proliferation by interfering with the process for glycolysis and gluconeogenesis. Based on the experimental result, we proposed a new mouthwash method to evaluate the anti-bacterial activity. Compared with AP, TP can inhibit the E. coli's growth within 2 h with a low concentration (0.5%) and a short dosing time (5 min). This study extends the applications of THDG and establishes a new assessment method for the pharmacology activity of Chinese herbal medicine.

Rapid synthesis of a highly active and uniform 3-dimensional SERS substrate for on-spot sensing of dopamine.

A rapid method is described for the preparation of a highly uniform and sensitive SERS substrate by an improved 'drop-and-dry' method. Gold nanobipyramids (Au NBPs) were prepared inside the nanoholes (nanowalls) of anodic aluminum oxide (AAO) templates with a typically 5-µm nanohole depth. The SERS substrate can be prepared by this method within 40 s and on large scale. The SERS signals obtained with this Au NBPs-AAO substrate is stronger by four-orders of magnitude compared to conventional a silicon wafer substrate. The SERS signal for dopamine (DA; measured at 1311 cm-1) is found to be enhanced by a factor of 2.2 × 108. The response to DA extends from 10 nM to 0.1 mM, and the limit of detection is 6.5 nM (at S/N = 3). The assay was applied to the determination of DA in spiked human serum. Graphical abstract Schematic presentation of a highly active and uniform 3-dimensional substrate composed of gold nanobipyramids and anodic aluminum oxide (Au NBP/AAO). It was used for on-spot sensing of dopamine.

Polysaccharides from Tetrastigma hemsleyanum Diels et Gilg: Extraction optimization, structural characterizations, antioxidant and antihyperlipidemic activities in hyperlipidemic mice.

In this study, a novel polysaccharide TDGP-3 from Tetrastigma hemsleyanum Diels et Gilg was extracted by enzymolysis-ultrasonic assisted extraction (EUAE) method and its antioxidant and hypolipidemic activities were evaluated. With a molecular weight of 3.31 × 105 Da, TDGP-3 was composed of 1,4-Glcp, 1,4-Glap and 1,3,6-Manp linkage in the main chain. TDGP-3 showed potential effects on relieving hyperlipidemia and preventing oxidative stress, reflecting by regulating blood lipid levels (TC, TG, HDL-C and LDL-C), decreasing the contents of MDA in liver, restoring the activities of hepatic antioxidant enzymes (SOD, GSH-Px and CAT). The results clearly indicated that the TDGP-3 possesses a potent hypolipidemic activity and might be used for hyperlipidaemia treatment.

Hypoglycemic Effects of a Polysaccharide from Tetrastigma hemsleyanum Diels & Gilg in Alloxan-Induced Diabetic Mice.

Tetrastigma hemsleyanum Diels & Gilg, a well-known traditional Chinese medicine, possesses antitumor and anti-inflammatory activity, etc. However, the anti-diabetic effect has not been determined. In our present study, a water-soluble polysaccharide, named THP with molecular weight of 93 307 Da, was isolated from T. hemsleyanum by DEAE-52 ion-exchange and Sephadex G-100 chromatography. It contains rhamnose, arabinose, mannose, glucose, and galactose in the molar ratio of 0.07:0.14:0.38:0.21:0.31. Then anti-diabetic effects of THP were examined by treating alloxan-induced diabetic mice with different doses (100, 200, and 300 mg/kg) of THP orally. The results showed that THP could decrease the blood glucose, TC, TG, LDL-C levels, increase the body weight, HDL-C, insulin levels, and enhance the activities of antioxidant enzyme system in alloxan-induced diabetic mice. Furthermore, the histopathological examination of pancreas, liver, and kidney indicated that THP could protect and reverse ß-cells in diabetic mice with low damage to liver and kidney, which suggests that THP may stimulate pancreatic release of insulin and can be an effectively potential candidate for diabetes mellitus.

Highly Selective and Sensitive Electrochemiluminescence Biosensor for p53 DNA Sequence Based on Nicking Endonuclease Assisted Target Recycling and Hyperbranched Rolling Circle Amplification.

An ultrasensitive and specific electrochemiluminescence (ECL) biosensor has been designed for the p53 DNA sequence, which is based on cascade signal amplification of nicking endonuclease assisted target recycling and hyperbranched rolling circle amplification (HRCA). First of all, biotin modified hairpin capture DNA (HP) probe was immobilized on the surface of streptavidin magnespheres paramagnetic particles (PMPs). Target DNA hybridized with the loop portion of the HP probe, therefore unfolding HP to form a double-stranded DNA (dsDNA) containing the specific nicking site of the nicking endonuclease. Then, the nicking endonuclease recognized the specific nicking site and cleaved the HP into two pieces, liberating target DNA and the complementary sequence piece for the padlock probe. The intact target DNA would initiate the next cycle of hybridization and cleavage, thereby releasing multiple complementary sequences for the padlock probes. The liberated complementary sequences hybridized with the padlock probes, subsequently inducing the HRCA reaction and generating numerous dsDNA segments. Herein, Ru(phen)3(2+) was embedded into dsDNA and worked as ECL signal reporter. The reaction products were eventually pretreated by dialysis tube with the cutoff membrane to remove the residual Ru(phen)3(2+) in the solution for the following ECL measurements. Using this cascade amplification strategy, an ultrasensitive p53 DNA sequence detection method was developed with a wide linear range from 0.05 to 100 fM and a low detection limit of 0.02 fM. Moreover, this cascade amplified ECL biosensor had specific recognition capacity for noncomplementary and single- and double-base mismatched DNA. The proposed ECL biosensor might have a great potential in biomedical research and clinic analysis.

Multicolor ELISA based on alkaline phosphatase-triggered growth of Au nanorods.

Seed-mediated synthesis of gold nanorods (AuNRs) has been widely used for diverse applications in the past decade. In this work, this synthetic process is demonstrated for multicolor biosensing for the first time. Our investigation reveals that ascorbic acid acts as a key factor to mediate the growth of AuNRs. This phenomenon is incorporated into the alkaline phosphatase (ALP)-enzyme-linked immunosorbent assay (ELISA) system based on the fact that ALP can catalyze the conversion of ascorbic acid-phosphate into ascorbic acid with high efficiency. This allows us to develop a multicolor ELISA approach for sensitive detection of disease biomarkers with the naked eye. We show the proof-of-concept multicolor ELISA for the detection of prostate-specific antigen (PSA) in human serum. The results show that different colors are presented in response to different concentrations of PSA, and a detection limit of 3 × 10(-15) g mL(-1) in human serum was achieved. The proposed multicolor ELISA could be a good supplement to conventional ELISA for POC diagnostics.