MiRNA-6870-3p Regulates Lipopolysaccharide Induced Epicardial Adipose Tissue Inflammatory Genes via Targeting Tollip-Mediated JNK and NF-κB Signaling in Coronary Artery Disease.

MiR-6870-3p acts as a crucial regulator of gene expression at the posttranscriptional level and participates in immune responses. However, the roles of miR-6870-3p and its target genes and their underlying mechanisms in the inflammatory responses of epicardial adipose tissues (EATs) are unknown.MiRNA microarray was used to collect the miRNA expression profiles of EATs from five patients with coronary artery disease (CAD) and four individuals without CAD (n-CAD). Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to examine the expression of miR-6870-3p in the EATs. The mRNA and protein expression levels of Tollip and the key genes of the Toll-like receptor 4 (TLR4) signaling pathway were examined by qRT-PCR and Western blot analysis. The levels of inflammatory factors in the cell supernatant were measured by enzyme-linked immunosorbent assay (ELISA). We used a dual-luciferase reporter assay to validate the target gene of miR-6870-3p. The protein expression levels of c-Jun N-terminal kinase (JNK) and nuclear factor kappa B (NF-κB) were measured by Western blot analysis.Our results showed that miR-6870-3p was higher in the CAD EATs than in the n-CAD EATs. MiR-6870-3p was positively correlated with TLR4, interleukin (IL)-6, JNK, NF-κB (p65), and tumor necrosis factor (TNF)-α in the CAD EAT samples. Lipopolysaccharide (LPS) treatment upregulated the miR-6870-3p expression and downregulated the Tollip expression in the macrophages. When the macrophages were stimulated with LPS, MiR-6870-3p upregulation also aggravated the production of proinflammatory cytokines. The result of the luciferase reporter assays confirmed that miR-6870-3p directly targets Tollip. Moreover, miR-6870-3p upregulation in the macrophages resulted in the activation of the JNK/NF-κB pathway.Our study showed that miR-6870-3p regulates human EAT inflammation by targeting the Tollip-mediated JNK and NF-κB signaling pathways.

[Interaction between homologous functional food Astragali Radix and intestinal flora].

Traditional Chinese medicine(TCM) is the treasure of our culture, and TCM theory is the core of traditional Chinese medicine. Many of its concepts can be unified and balanced with modern functional food ideas. Even in ancient days, people had already found that medicine and food have the same source. Nowadays, homology between drug and food has been accepted widely. Astragali Radix and some other herbs have been used both as food and medicine, with a variety of bio-active substances, so such herbs can be used as characteristics resources to be developed into functional food. It's a combination of traditional medicine and modern ideas. Flavonoids, polysaccharides and saponins, the main compositions of Astragali Radix, can keep intestinal microenvironment homeostasis and human health by influencing the population structure, metabolism and intestinal cell function of intestinal flora. On the other hand, intestinal flora is also involved in the absorption, metabolism, transformation and other steps of these active ingredients in the body, which has an impact on their effectiveness and improves their bioavailability, playing an essential role in the relevant mechanism of their effectiveness. In this paper, we summarize the interaction between the above three functional ingredients in Astragali Radix and intestinal flora, sum up the interaction between these three functional ingredients of other homologous drugs and intestinal flora, provide a theoretical basis for the mechanism and application of functional food materials, and propose some suggestions and prospects for their future development.

Toll-like receptor 3-activated macrophages confer anti-HCV activity to hepatocytes through exosomes.

Exosomes are a class of cell-released small vesicles that mediate intercellular communication by delivering functional factors to recipient cells. During hepatitis C virus (HCV) infection, the interaction between liver resident macrophages and hepatocytes is a key component in liver innate immunity. In this study, we explored the role of exosomes in the delivery of innate anti-HCV factors to hepatocytes from macrophages. We showed that supernatant from TLR3-activated macrophage cultures could efficiently inhibit HCV replication in Huh7 cells. This macrophage-mediated anti-HCV activity was through exosomes because inhibiting exosomes could abrogate the action of macrophages. Further analyses demonstrated that TLR3-activated macrophages release exosomes that contain anti-HCV microRNA (miRNA)-29 family members. Inhibiting miRNA29 could restore HCV replication. These findings suggest a novel antiviral mechanism in liver innate immunity against HCV infection and provide insights to support further studies on developing exosome-based delivery system for disease treatment.-Zhou, Y., Wang, X., Sun, L., Zhou, L., Ma, T.-C., Song, L., Wu, J.-G., Li, J.-L., Ho, W.-Z. Toll-like receptor 3-activated macrophages confer anti-HCV activity to hepatocytes through exosomes.

Activation of TLR3/interferon signaling pathway by bluetongue virus results in HIV inhibition in macrophages.

Bluetongue virus (BTV), a nonenveloped double-stranded RNA virus, is a potent inducer of type Ι interferons in multiple cell systems. In this study, we report that BTV16 treatment of primary human macrophages induced both type I and III IFN expression, resulting in the production of multiple antiviral factors, including myxovirus resistance protein A, 2',5'-oligoadenylate synthetase, and the IFN-stimulated gene 56. Additionally, BTV-treated macrophages expressed increased HIV restriction factors (apolipoprotein B mRNA-editing enzyme catalytic polypeptide 3 G/F/H) and CC chemokines (macrophage inflammatory protein 1-α, macrophage inflammatory protein 1-ß, regulated on activation of normal T cell expressed and secreted), the ligands for HIV entry coreceptor CC chemokine receptor type 5. BTV16 also induced the expression of tetherin, which restricts HIV release from infected cells. Furthermore, TLR3 signaling of macrophages by BTV16 resulted in the induction of several anti-HIV microRNAs (miRNA-28, -29a, -125b, -150, -223, and -382). More importantly, the induction of antiviral responses by BTV resulted in significant suppression of HIV in macrophages. These findings demonstrate the potential of BTV-mediated TLR3 activation in macrophage innate immunity against HIV.

Quality evaluation of the leaves of Magnolia officinalis var. biloba using high-performance liquid chromatography fingerprint analysis of phenolic compounds.

The high-performance liquid chromatography fingerprint method is a simple and reliable technique to evaluate the quality of leaves of Magnolia officinalis Rehd.et Wils. var. biloba Rehd.et Wils. We used the following bioactive phenolic constituents as reference compounds: rutin, afzelin, hyperoside, isoquercitrin, quercetin-3-O-α-l-rhamnoside, honokiol and magnolol. The conditions of an Agilent 1200 HPLC were: YMC-Pack-ODS-AQ column (250 × 4.6 mm id S-5 µm, 12 nm), mobile phase acetonitrile and 0.2% phosphoric acid in a gradient elute mode, flow rate 1.0 mL/min, detection wavelength 280 nm and column temperature 30°C. The analytical method was validated in terms of linearity, stability, repeatability, precision and recovery tests. While performing fingerprint analysis, we identified 11 peaks as characteristic peaks and assessed the similarities of 17 samples collected from different geological regions of China. The peak areas were used to evaluate the variation in the chemical composition of the tested samples. For this purpose, we performed hierarchical cluster analysis of the peak areas. Our results indicate that simultaneous determination of multiple ingredients could be done through chromatographic fingerprint analysis. Therefore, this high-performance liquid chromatography fingerprint method was readily utilized to evaluate the quality of leaves of M. officinalis var.biloba, which are used in several traditional herbal preparations.

QTL mapping based on the embryo and maternal genetic systems for non-essential amino acids in rapeseed (Brassica napus L.) meal.

BACKGROUND: Non-essential amino acids are a good source of nitrogen and also very important contributors to the metabolic process. Analysis of quantitative trait locus (QTL) simultaneously located on the amphidiploid embryo and maternal plant nuclear genomes for non-essential amino acid contents in rapeseed meal across different environments was conducive to further clarify the genetic mechanism of seed quality traits. RESULTS: Twenty-eight QTLs associated with arginine (five QTLs), histidine (four QTLs), glutamic acid (three QTLs), glycine (three QTLs), proline (three QTLs), alanine (four QTLs) and aspartic acid (six QTLs) contents were identified in present study. All of these QTLs had significant additive main effects from embryo and maternal plant nuclear genomes with eight of them showing significant embryo dominance main effects and 12 showing notable QTL × environment interaction effects. Among them, 12 QTLs were major QTLs which could explain 13.27-35.71% of the phenotypic variation. Specially, five QTL clusters associated with several QTLs related to multiple traits were distributed on chromosomes A1, A4, A5, A7 and C2. CONCLUSION: Non-essential amino acids in rapeseed meal could be simultaneously controlled by the genetic effects from the QTLs which were located on the chromosomes both in the embryo and maternal plant genetic systems.

Hepatoprotective effect of ganoderma triterpenoids against oxidative damage induced by tert-butyl hydroperoxide in human hepatic HepG2 cells.

CONTEXT: Ganoderma triterpenoids (GTs) have been recognised as an important bioactive ingredient in Ganoderma Lucidum (Leyss. ex Fr.) Karst. (Polyporaceae), widely used for treating and preventing chronic hepatopathy of various etiologies. OBJECTIVE: The objective of this study is to better understand the hepatoprotective effect of GTs and to enhance their use in food supplement pharmaceutical and medical industries. MATERIALS AND METHODS: HepG2 cells were pretreated in the presence or absence of GTs (50, 100 and 200 µg/ml) for 4 h, then exposed with 60 µmol/L of t-BHP for an additional 4 h. The cell viability was evaluated by MTT method. ALT, AST and LDH production in culture medium and intracellular MDA, GSH and SOD levels were determined. Moreover, the total triterpenoid content and chemical constituents in GTs were detected by ultraviolet spectrophotometry and HPLC/Q-TOF-MS, respectively. RESULTS: GTs (50, 100 and 200 µg/ml) significantly increased the relative cell viability by 4.66, 7.78 and 13.46%, respectively, and reduced the level of ALT by 11.44%, 33.41% and 51.24%, AST by 10.05%, 15.63% and 33.64%, and LDH by 16.03%, 23.4% and 24.07% in culture medium, respectively. GTs could also remarkably decrease the level of MDA and increase the content of GSH and SOD in HepG2 cells. Furthermore, the total triterpenoid content in GTs was 438 mg GAAEs/g GTs. And 16 triterpenoids in GTs were identified or tentatively characterised. DISCUSSION AND CONCLUSION: Our results showed that GTs had potent cytoprotective effect against oxidative damage induced by t-BHP in HepG2 cells, thus suggesting their potential use as liver protectant.

Biometabolic Distribution of 99mTc-3PRGD2 and Its Potential Value in Monitoring Chemotherapeutic Effects.

Previous studies have reported that 99mTc-3PRGD2 is an excellent tumor imaging agent that showed a good correlation with integrin αvß3, a main factor of tumor-induced angiogenesis. In this study, we investigated the biometabolic distribution characteristics of 99mTc-3PRGD2 with a continuous dynamic acquisition mode to explore the potential value of 99mTc-3PRGD2 in monitoring chemotherapeutic effects in VX2 tumor models. Eighteen rabbits with 27 implanted VX2 squamous cell tumors were randomly divided into a nontreated control group (NTG, n = 8; 12 tumors) and a treatment group (TG, n = 10; 15 tumors). 99mTc-3PRGD2 imaging was performed prior to cisplatin injection and repeated on days 0, 1, 7, and 14 postinjection. Continuous dynamic scanning up to 30 minutes; static imaging at 0.5 hours, 1 hour, and 3 hours; and single-photon emission computed tomography/computed tomography (SPECT/CT)-integrated imaging at 3 hours post-99mTc-3PRGD2 injection were performed. The peak time (time to reach peak in dynamic curve), tumor to normal (T/N) ratios, and their change rates relative to pretherapy were calculated. Autoradiography, hematoxylin-eosin (H&E) staining, and CD31 and integrin αv immunohistochemical staining were examined. VX2 tumors were clearly visualized at 3 hours post-99mTc-3PRGD2 injection. Tumors in the TG shrank significantly on day 7 after cisplatin administration (p < .05). The half-life (t1/2) of the radiotracer in heart, liver, and tumor in the NTG were 3.43 ± 0.94 minutes, 13.41 ± 9.17 minutes, and 70.83 ± 33.37 minutes, respectively. The peak time was delayed in the TG immediately and continuously after cisplatin administration compared to the peak time in the NTG. The T/N values and their change rates decreased significantly in the TG compared to the NTG after therapy (p < .05). The immunostained areas were significantly decreased in the TG (p < .05) compared to the NTG. 99mTc-3PRGD2 was an excellent imaging agent for demonstrating tumor angiogenesis. The peak time, T/N values, and their change rates were sensitive parameters to monitor early chemotherapeutic effects. Due to the specific target mechanism and the cost-effective value of 99mTc-3PRGD2, 99mTc-3PRGD2 SPECT imaging may have potential in detecting the therapeutic effects of anticancer therapy.

Induction of interferon-λ contributes to TLR3 and RIG-I activation-mediated inhibition of herpes simplex virus type 2 replication in human cervical epithelial cells.

STUDY HYPOTHESIS: Is it possible to immunologically activate human cervical epithelial cells to produce antiviral factors that inhibit herpes simplex virus type 2 (HSV-2) replication? STUDY FINDING: Our results indicate that human cervical epithelial cells possess a functional TLR3/RIG-I signaling system, the activation of which can mount an Interferon-λ (IFN-λ)-mediated anti-HSV-2 response. WHAT IS KNOWN ALREADY: There is limited information about the role of cervical epithelial cells in genital innate immunity against HSV-2 infection. STUDY DESIGN, SAMPLES/MATERIALS, METHODS: We examined the expression of toll-like receptors (TLRs) and retinoic acid-inducible I (RIG-I) in End1/E6E7 cells by real-time PCR. The IFN-λ induced by TLR3 and RIG-I activation of End1/E6E7 cells was also examined by real-time PCR and ELISA. HSV-2 infection of End1/E6E7 cells was evaluated by the real-time PCR detection of HSV-2 gD expression. The antibody to IL-10Rß was used to determine whether IFN-λ contributes to TLR3/RIG-I mediated HSV-2 inhibition. Expression of interferon regulatory factor 3 (IRF3), IRF7, IFN-stimulated gene 56 (ISG56), 2'-5'-oligoadenylate synthetase I (OAS-1) and myxovirus resistance A (MxA) were determined by the real-time PCR and western blot. End1/E6E7 cells were transfected with shRNA to knockdown the IRF3, IRF7 or RIG-I expression. Student's t-test and post Newman-Keuls test were used to analyze stabilized differences in the immunological parameters above between TLR3/RIG-I-activated cells and control cells. MAIN RESULTS AND THE ROLE OF CHANCE: Human cervical epithelial cells expressed functional TLR3 and RIG-I, which could be activated by poly I:C and 5'ppp double-strand RNAs (5'ppp dsRNA), resulting in the induction of endogenous interferon lambda (IFN-λ). The induced IFN-λ contributed to TLR3/RIG-I-mediated inhibition of HSV-2 replication in human cervical epithelial cells, as an antibody to IL-10Rß, an IFN-λ receptor subunit, could compromise TLR3/RIG-I-mediated inhibition of HSV-2. Further studies showed that TLR3/RIG-I signaling in the cervical epithelial cells by dsRNA induced the expression of the IFN-stimulated genes (ISGs), ISG56, 2'-5'-oligoadenylate synthetase I (OAS-1) and myxovirus resistance A (MxA), the key antiviral elements in the IFN signaling pathway. In addition, we observed that the topical treatment of genital mucosa with poly I:C could protect mice from genital HSV-2 infection. LIMITATIONS, REASONS FOR CAUTION: Future prospective studies with primary cells and suitable animal models are needed in order to confirm these outcomes. WIDER IMPLICATIONS OF THE FINDINGS: The findings provide direct and compelling evidence that there is intracellular expression and regulation of IFN-λ in human cervical epithelial cells, which may have a key role in the innate genital protection against viral infections. LARGE SCALE DATA: Not applicable. STUDY FUNDING AND COMPETING INTERESTS: This work was supported by the National Natural Science Foundation of China (81301428 to L.Z. and 81271334 to W.-Z.H.), the Fundamental Research Funds for the Central Universities (2042015kf0188 to L.Z.), the China Postdoctoral Science Foundation (2013M531745 to L.Z.), the Development Program of China ('973', 2012CB518900 to W.-Z.H.) from the Ministry of Science and Technology of the People's Republic of China, grants (DA12815 and DA022177 to W.-Z.H.) from the National Institute on Drug Abuse (NIDA) and the open project of Hubei Key Laboratory of Wudang Local Chinese Medicine Research (WDCM005 to M.S.). The authors declare no competing financial interests.

Rice grassy stunt virus nonstructural protein p5 serves as a viral suppressor of RNA silencing and interacts with nonstructural protein p3.

Rice grassy stunt virus (RGSV), a member of the genus Tenuivirus, causes serious rice disease in Southeast Asian countries. In this study, a green fluorescent protein (GFP)-based transient expression assay was conducted to show that p5, encoded on RNA5 in the viral sense, is a viral suppressor of RNA silencing (VSR). Protein-protein interactions (PPIs) between p5 and all RGSV proteins except pC1 and pC2 were investigated using Gal4-based yeast two-hybrid (Y2H) experiments. The results demonstrated that p5 interacts with itself and with p3 encoded on RNA3 in the viral sense. p5-p5 and p5-p3 interactions were detected by bimolecular fluorescence complementation (BiFC) assay, and the p5-p3 interaction was confirmed by subcellular co-localization and co-immunoprecipitation (Co-IP) assays. Using the Y2H system, we demonstrated that the p5-p3 interaction requires both the N-terminal (amino acid residues 1 to 99) and C-terminal (amino acid residues 94 to 191) domains of p5. In addition, either p5 or p3 could enhance the pathogenicity of potato virus X (PVX) in Nicotiana benthamiana plants. A much more significant enhancement of PVX pathogenicity and accumulation was observed when p5 and p3 were expressed together. Our data also showed that RGSV p3 does not function as a VSR, and it had no effect on the VSR activity of p5 or the subcellular localization pattern of p5 in plant cells from Nicotiana benthamiana.

MiR-125b promotes cell migration and invasion by targeting PPP1CA-Rb signal pathways in gastric cancer, resulting in a poor prognosis.

BACKGROUND: MiR-125b functions as an oncogene in many cancers; however, its clinical significance and molecular mechanism in gastric cancers have never been sufficiently investigated. Here, we elucidated the functions and molecular regulated pathways of MiR-125b in gastric cancer. METHODS: We investigated MiR-125b expression in fresh tissues from 50 gastric cancer patients and 6 gastric cancer cell lines using RT-PCR, and explored its prognostic value by hybridizing MiR-125b in situ for 300 clinical gastric tumor tissues with pathological diagnosis and clinical parameters. The effects of MiR-125b on gastric cancer cells and downstream target genes and proteins were analyzed by MTT, transwell assay, RT-PCR, and western blot on the basis of silencing MiR-125b in vitro. Luciferase reporter plasmid was constructed to demonstrate MiR-125b's direct target. RESULTS: MiR-125b was upregulated in gastric cancer tissues and cell lines, and significantly promoted cellular proliferation, migration, and invasion by downregulating the expression of PPP1CA and upregulating Rb phosphorylation. MiR-125b expression was significantly correlated with tumor size and depth of invasion, lymph nodes, distant metastasis, and TNM stage. The high-MiR-125b-expression group had a significantly poorer prognosis than the low-expression group (P < 0.05) in stages I, II, and III, and the 5-year survival rate in of the high-expression group was significantly lower than that of the low-expression group. CONCLUSIONS: MiR-125b functions as an oncogene by targeting downregulated PPP1CA and upregulated Rb phosphorylation in gastric cancer. MiR-125b not only promotes cellular proliferation, migration, and invasion in vitro, but also acts as an independent prognostic factor in gastric cancer.

Self-retaining contact lens simplifies intravitreal injection course in rats.

BACKGROUND: The aim of this study was to design a self-retaining rat contact lens to simplify intravitreous injection in rats. MATERIAL/METHODS: A self-retaining, plane-concave prism contact lens customized for rats was designed. Forty diabetic rats were randomly divided into 2 groups and received the intravitreous injection of 10 µl of a cell suspension containing bone marrow-derived stroma cell (BMSC). Group A: used a microsyringe and a rat contact lens (n=20). Group B: used the same microsyringe and a traditional cover-slip (n=20).The duration of the intravitreous injection course and the success rate of intravitreous injection were observed. RESULTS: With the use of a self-retaining rat contact lens, a clear and stable view of the rat fundus was provided and the intravitreous injection course of rats quickly achieved, averaging 4.65±0.53 min in Group A and 12.33±2.79 min in Group B. The difference was statistically significant, and the time saved averaged 7.68 min. None of the Group A rats had retinal bleeding or lens injury; whereas 2 of 20 Group B rats had bleeding and 1 of 20 had lens injury. There was no significant difference between the rats in Group A and Group B. CONCLUSIONS: A self-retaining rat contact lens is a potentially powerful instrument that allows high-quality observation of the rat fundus and simplifies the course of intravitreal injection.

Quantitative and chemical fingerprint analysis for the quality evaluation of Receptaculum Nelumbinis by RP-HPLC coupled with hierarchical clustering analysis.

A simple and reliable method of high-performance liquid chromatography with photodiode array detection (HPLC-DAD) was developed to evaluate the quality of Receptaculum Nelumbinis (dried receptacle of Nelumbo nucifera) through establishing chromatographic fingerprint and simultaneous determination of five flavonol glycosides, including hyperoside, isoquercitrin, quercetin-3-O-ß-d-glucuronide, isorhamnetin-3-O-ß-d-galactoside and syringetin-3-O-ß-d-glucoside. In quantitative analysis, the five components showed good regression (R > 0.9998) within linear ranges, and their recoveries were in the range of 98.31%-100.32%. In the chromatographic fingerprint, twelve peaks were selected as the characteristic peaks to assess the similarities of different samples collected from different origins in China according to the State Food and Drug Administration (SFDA) requirements. Furthermore, hierarchical cluster analysis (HCA) was also applied to evaluate the variation of chemical components among different sources of Receptaculum Nelumbinis in China. This study indicated that the combination of quantitative and chromatographic fingerprint analysis can be readily utilized as a quality control method for Receptaculum Nelumbinis and its related traditional Chinese medicinal preparations.

[Promotion of tuition quality in Genetics by developing an excellent instructional system].

This article, which is based on the requirements for developing the course of Genetics as an outstanding course, summarizes the experience of engendering an excellent instructional system, expatiates on the effects of its application to the teaching process of "Genetics" and next step work for continuing this instructional system. The course quality of "Genetics" has been improved under the excellent instructional system by including different teaching methods, renovating the teaching contents, innovating the teaching means, developing a practical courseware, writing extractive textbooks, reforming experimental teaching, and constructing an instructional network, together with teaching methods related to scientific research.

[Morphological characteristics and gene mapping of a novel bent pedicel branch (bpb1) mutant in rice].

Rice pedicels are tightly associated with the yield of grain. In the present study, a novel and stable pedicel mutant bpb1 (bent pedicel branch 1) was obtained from the wild type "Zhenong 7" after 60Co γ-ray treatment. The mutant had the typical phenotype of bent pedicel branches with multiple abnormal phenotypes, such as longer pedicels, short panicles, and dwarfism. Detail examination using scanning electron microscopy revealed that the pedicel epidermal hairs and stomas in the mutant were smaller than those in the wild type. The epidermal and sclerenchymatous cells were arranged irregularly, and the cells in the bend region of pedicels became smaller and arranged closely. The transverse observation of the mutant pedicel branches showed that the small vascular bundles arranged differently from those of the wild type. Genetic analysis indicated that the abnormal phenotypes were controlled by a single recessive gene. Using the F2 mapping population from the bpb1 mutant crossed with the japonica rice variety "Zhenongda 104", the bpb1 gene was mapped in a 343 kb region between two SSR markers, RM21537 and RM21552, at the long arm of chromosome 7. Because no homologous gene was found in this region until now, bpb1 might be a novel gene related to the pedicel development and growth. This study could be beneficial to future cloning and functional analysis of the bpb1 gene.

[Acupuncture for glaucoma-induced optic atrophy: a randomized controlled trial].

OBJECTIVE: To observe the clinical effect of acupuncture for glaucoma-induced optic atrophy. METHODS: A total of 70 patients (89 affected eyes) with glaucoma-induced optic atrophy were randomized into an observation group and a control group, 35 cases in each group. The control group was given basic western medicine treatment. In the observation group, on the basis of the treatment in the control group, acupuncture was applied at main acupoints i.e. Baihui (GV 20), Shangjingming (Extra), Chengqi (ST 1), Fengchi (GB 20), Zusanli (ST 36), combined with supplementary acupoints based on syndrome differentiation, once every three days, twice a week. The treatment for 3 months was required in both groups. Before treatment, after treatment and in follow-up of 6 months after treatment, the best corrected visual acuity (BCVA), intraocular pressure (IOP), indexes of visual field (visual field index [VFI], mean deviation [MD], pattern standard deviation [PSD]) and mean thickness of retinal nerve fiber layer (RNFL) were observed in the two groups. RESULTS: Compared before treatment, BCVA was decreased after treatment and in follow-up in the control group (P<0.05); in the follow-up, BCVA in the observation group was higher than that in the control group (P<0.05). On each time point before and after treatment, there was no significant difference within or between the two groups (P>0.05). After treatment and in the follow-up, the mean thickness of RNFL was larger than the control group (P<0.05). CONCLUSION: On the basis of the basic western medicine treatment, acupuncture can delay the decline of vision and the thinning of retinal nerve fiber layer in patients with glaucoma-induced optic atrophy.

A mutation screening platform for rapeseed (Brassica napus L.) and the detection of sinapine biosynthesis mutants.

We developed two mutant populations of oilseed rape (Brassica napus L.) using EMS (ethylmethanesulfonate) as a mutagen. The populations were derived from the spring type line YN01-429 and the winter type cultivar Express 617 encompassing 5,361 and 3,488 M(2) plants, respectively. A high-throughput screening protocol was established based on a two-dimensional 8× pooling strategy. Genes of the sinapine biosynthesis pathway were chosen for determining the mutation frequencies and for creating novel genetic variation for rapeseed breeding. The extraction meal of oilseed rape is a rich protein source containing about 40% protein. Its use as an animal feed or human food, however, is limited by antinutritive compounds like sinapine. The targeting-induced local lesions in genomes (TILLING) strategy was applied to identify mutations of major genes of the sinapine biosynthesis pathway. We constructed locus-specific primers for several TILLING amplicons of two sinapine synthesis genes, BnaX.SGT and BnaX.REF1, covering 80-90% of the coding sequences. Screening of both populations revealed 229 and 341 mutations within the BnaX.SGT sequences (135 missense and 13 nonsense mutations) and the BnaX.REF1 sequences (162 missense, 3 nonsense, 8 splice site mutations), respectively. These mutants provide a new resource for breeding low-sinapine oilseed rape. The frequencies of missense and nonsense mutations corresponded to the frequencies of the target codons. Mutation frequencies ranged from 1/12 to 1/22 kb for the Express 617 population and from 1/27 to 1/60 kb for the YN01-429 population. Our TILLING resource is publicly available. Due to the high mutation frequencies in combination with an 8× pooling strategy, mutants can be routinely identified in a cost-efficient manner. However, primers have to be carefully designed to amplify single sequences from the polyploid rapeseed genome.

A simultaneous iris angiography technique in pigmented rabbits.

PURPOSE: To report a simultaneous iris angiography (IA) technique combined with the use of indocyanine green (ICG) and fluorescein sodium (FS) in pigmented rabbits. METHODS: 15 rabbits were randomly divided into 3 groups according to the dye doses: the dose of the lower-volume group was 2 mg FS and 2 mg ICG; the dose of the moderate-volume group was 5 mg FS and 5 mg ICG, and the dose of the higher-volume group was 8 mg FS and 8 mg ICG. Fifteen IAs were performed by a confocal scanning laser ophthalmoscope and a rapid localization technique giving a flat-on image. RESULTS: Simultaneous digital angiography provided two kinds of a fully dynamic video of iris vessels fluorescein in all pigmented rabbits, which is the same in pigmented and albino rabbits and no vascular pattern was hidden by the iris stromal pigment. Furthermore, dye doses influenced the IA effect in rabbits. The lower-volume dye can demonstrate iris vessels, without obvious leakage, which, however, extinguish rapidly after 10-15 s. In the higher-volume group, vascular imaging lasted longer, but subsequently significant FS and ICG leakage appeared as streams on the surface of the iris until the dye disappeared from the iris vessels; the diffuse and intense aqueous fluid lasted 1 day. The moderate-volume dye displayed vessels clearly by ICG for 300 s, without leakage; FS clear vessels were maintained for about 15-20 s and the structure of the iris became fuzzy due to quick leakage. CONCLUSIONS: Simultaneous IA with a rapid localization technique allows high-quality imaging of the pigmented rabbits and the moderate dosage of 5 mg FS and 5 mg ICG is preferred for best visualization.

Antioxidant activities of extract and fractions from receptaculum nelumbinis and related flavonol glycosides.

The antioxidant activities of ethanolic crude extract (ECE) and its four different solvent sub-fractions (namely, petroleum ether fraction (PEF), ethyl acetate fraction (EAF), n-butanol fraction (BF) and the aqueous fraction (AF) from the receptacles of Nelumbo nucifera Gaertn. (Receptaculum Nelumbinis) were investigated using two in vitro antioxidant assays. BF showed the highest total phenolic content (607.6 mg/g gallic acid equivalents), total flavonoid content (862.7 mg/g rutin equivalents) and total proanthocyanidin content (331.0 mg/g catechin equivalents), accompanied with the highest antioxidant activity compared to other fractions through 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging assays. Five flavonol glycosides, namely hyperoside (1), isoquercitrin (2), quercetin-3-O-ß-d-glucuronide (3), isorhamnetin-3-O-ß-d-galactoside (4) and syringetin-3-O-ß-d-glucoside (5) were isolated from the Receptaculum Nelumbinis. Compounds 2-5 were isolated for the first time from the Receptaculum Nelumbinis. The five isolated flavone glycosides, particularly compounds 1-3, demonstrated significant DPPH and ABTS radical scavenging activity, with IC(50) values of 8.9 ± 0.2, 5.2 ± 0.2, 7.5 ± 0.1 for DPPH and 114.2 ± 1.7, 112.8 ± 0.8, 172.5 ± 0.7 µg/mL for ABTS, respectively. These results suggest that Receptaculum Nelumbinis has strong antioxidant potential and may be potentially used as a safe and inexpensive bioactive source of natural antioxidants.