Gypenoside Inhibits Bovine Viral Diarrhea Virus Replication by Interfering with Viral Attachment and Internalization and Activating Apoptosis of Infected Cells.

Bovine viral diarrhea virus (BVDV) causes a severe threat to the cattle industry due to ineffective control measures. Gypenoside is the primary component of Gynostemma pentaphyllum, which has potential medicinal value and has been widely applied as a food additive and herbal supplement. However, little is known about the antiviral effects of gypenoside. The present study aimed to explore the antiviral activities of gypenoside against BVDV infection. The inhibitory activity of gypenoside against BVDV was assessed by using virus titration and performing Western blotting, quantitative reverse transcription PCR (RT-qPCR), and immunofluorescence assays in MDBK cells. We found that gypenoside exhibited high anti-BVDV activity by interfering with the viral attachment to and internalization in cells. The study showed that BVDV infection inhibits apoptosis of infected cells from escaping the innate defense of host cells. Our data further demonstrated that gypenoside inhibited BVDV infection by electively activating the apoptosis of BVDV-infected cells for execution, as evidenced by the regulation of the expression of the apoptosis-related protein, promotion of caspase-3 activation, and display of positive TUNEL staining; no toxicity was observed in non-infected cells. Collectively, the data identified that gypenoside exerts an anti-BVDV-infection role by inhibiting viral attachment and internalization and selectively purging virally infected cells. Therefore, our study will contribute to the development of a novel prophylactic and therapeutic strategy against BVDV infection.

Synthesis and characterization of magnesium oxide nanoparticle-containing biochar composites for efficient phosphorus removal from aqueous solution.

The effective removal and recovery of phosphorus from aquatic environments are highly important for successful eutrophication control and phosphorus recycling. Herein, we prepared biochar containing MgO nanoparticles (MgO-biochar) by fast pyrolysis of MgCl2-impregnated corn stalks, probed its phosphate adsorption performance. Through the fast pyrolysis, the MgCl2 promoted the formation of micropores and mesoporous, and decomposed into MgO nanoparticles with the size smaller than 100 nm. The adsorption experiments showed that the adsorption property increased with the increase of Mg content, and had a strong correlation with the external surface area. And the phosphate adsorption was well described by the Langmuir-Freundlich model (maximum adsorption capacity was determined as 60.95 mg P/g). Kinetic analysis and characterization analysis of MgO-biochar for different adsorption time indicated that phosphate adsorption onto MgO-biochar was mainly controlled by rapid binding to the external surface (about 75% of the equilibrium adsorption amount), and the uptake rate was limited by the slow diffusion of phosphate into the biochar interior (about 25% of the equilibrium adsorption amount). The results suggested that the synthesized MgO-biochar with enough MgO active site dispersed on a higher external surface can be used as a potential adsorbent for phosphate removal and recovery from aqueous solution.

Response of alternative splice isoforms of OsRad9 gene from Oryza sativa to environmental stress.

Rad9 protein plays an important role in cell-cycle checkpoint signal transduction in human and yeast cells, but knowledge about Rad9 in plants is limited. This study reports that the Rad9 gene of rice can generate the transcript products OsRad9.1 and OsRad9.2 through alternative splicing. OsRad9.1, with all nine exons, is the main cell-cycle checkpoint protein involved in the response of rice to genotoxic stresses (ultraviolet radiation and antibiotic stress), environmental stresses (drought, salt, and heavy metal stress), and auxin stimuli (2,4-D, IAA, and IBA). Meanwhile, transcript isoform OsRad9.2, which lost exon7 and exon8, showed different preferential stimulation effects on these stresses and pollen development duration. These results might indicat that besides the monitoring and repair of DNA damage, Rad9 might involve in the development of pollen.

gp85 protein vaccine adjuvanted with silica nanoparticles against ALV-J in chickens.

This study focused on the effect of silica nanoparticles as adjuvant for vaccine applications comprised of gp85, a dominating structural protein of J Subgroup Avian Leukosis Virus (ALV-J), and which was evaluated by comparing with the responsiveness induced by that emulsified in Freund adjuvant. Thirty-six chickens were inoculated twice with gp85 adjuvanted with the silica nanoparticles or Freund's adjuvant at the 2nd and 3rd week old. Two weeks later, the inoculated chickens were challenged with a 102.2 50% tissue culture infective dose (TCID50) of ALV-J. The blood samples were collected weekly to detect the serum antibodies and viremia. Results showed that positive serum antibodies (S/P value>0.6) against gp85 emerged at the third week in the inoculated chickens, while the antibodies level persisted longer in silica nanoparticles adjuvanted-group to Freund's adjuvanted-group. Furthermore, viremia in silica nanoparticles adjuvanted-group was recovered more quickly compared with Freund's adjuvanted-group. Hence our study revealed that silica nanoparticles can effectively improve the protection of gp85 vaccine against ALV-J and present a better performance than Freund's adjuvant.

Selenium Administration Alleviates Toxicity of Chromium(VI) in the Chicken Brain.

Selenium (Se) can play a protective role against heavy metal toxicity. This experiment aims to evaluate the effect of Se supplementation at different doses on the chicken brains. Oxidative stress was induced in the chicken brains by chromium(VI). A total of 105 Hyland brown male chickens were randomly divided into seven groups, including the control group, poisoned group [6%LD50 K2Cr2O7 body weight (B.W.)], and detoxification groups K2Cr2O7 (6%LD50) + Se (0.31, 0.63, 1.25, 2.50, and 5.00 Na2SeO3 mg/kg B.W.) orally in water for 42 days. The chickens were detected by the activities of mitochondrial membrane potential, 2'-benzoyloxycinnamaldehyde, superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), and Ca2+-ATPase. Cr(VI) administration caused histopathological damage. In addition, changes in oxidative stress indicators were observed in the chicken's brains. Se supplement increased the levels of GSH, mitochondrial membrane potential (MMP), and Ca2+-ATPase and reduced MDA activity in the detoxification groups. However, the high-dose Se supplementation groups of 2.50 and 5.00 mg/kg reduced the activities of GSH, MMP, and Ca2+-ATPase; increased the brain-body ratio; and increased SOD activity. In conclusion, Cr(VI) exposure caused oxidative stress. Se exerted a remission effect on toxic responses in the chicken brains. However, a high Se concentration was synergistic to the toxic effect of Cr(VI).

Resveratrol attenuates inflammation and oxidative stress in epididymal white adipose tissue: implications for its involvement in improving steroidogenesis in diet-induced obese mice.

Chronic, low-grade systemic inflammation has been shown to play an important role in the development of obesity-related complications. Epididymal white adipose tissue (WAT) can influence testicular function through its endocrine function. The purpose of this study was to assess the effects of resveratrol on the epididymal WAT inflammatory response and on testicular steroidogenesis in obese individuals. Seven-week-old male C57BL/6J mice were fed a high-calorie and high-cholesterol diet (HCD group) or HCD supplemented with resveratrol (HCD+Res group) for 18 weeks. As we previously showed that resveratrol protects against Leydig cell steroidogenesis in HCD-induced obese mice, this study assessed macrophage infiltration in fat depots by measuring crown-like structure (CLS) density. Histological analysis showed that adipocyte size was significantly smaller and CLSs were less numerous in the HCD+Res group than the HCD group (P < 0.01). Additionally, resveratrol supplementation decreased Nfkb1 expression (P < 0.01) and increased the IκB-α protein abundance (P < 0.01) in epididymal WAT. Consistent with this alteration in NF-κB signaling, the expression of two classic proinflammatory cytokines, TNF-α (Tnfa) and IL-1ß (Il1b), were significantly decreased in the HCD+Res group compared with the HCD group (P < 0.01). Significant differences were also found in the expression of sirtuin1 (Sirt1) (P < 0.01) and manganese superoxide dismutase (Sod2) (P < 0.01) between the HCD and HCD+Res groups. Our data suggest that resveratrol can attenuate obesity-induced inflammation and oxidative stress in epididymal WAT, which partly accounts for its beneficial effects in testicular steroidogenesis.

Identification of novel acyl-ACP thioesterase gene ClFATB1 from Cinnamomum longepaniculatum.

A putative fatty acyl-acyl carrier protein (acyl-ACP) thioesterase (thioesterase) full-length cDNA sequence named as ClFATB1 was obtained from the seed cDNA library of Cinnamomum longepaniculatum by the SMART-RACE method. The novel gene encodes a protein of 382 amino acid residues with close homology to fatty acid thioesterase type B (FATB) enzymes of other plants, with two essential residues (His285 and Cys320) for thioesterase catalytic activity. The gene was transcribed in all tissues of C. longepaniculatum, the highest being in seeds. Recombinant ClFATB1 in Escherichia coli had higher specific activities against saturated 16:0- and 18:0-ACPs than on unsaturated 18:1-ACP. Overexpression of ClFATB1 in transgenic tobaccos upregulated thioesterase activities of crude proteins against 16:0-ACP and 18:0-ACP by 20.3 and 5.7%, respectively, and resulted in an increase in the contents of palmitic and stearic acids by 15.4 and 10.5%, respectively. However, ectopic expression of this gene decreased the substrate specificities of crude proteins to unsaturated 18:1-ACP by 12.7% in transgenic tobacco and lowered the contents of oleic, linoleic, and linolenic acids in transgenic leaves. So ClFATB1 would potentially upregulate the synthesis of saturated fatty acids and downregulate unsaturated ones in the fatty acid synthesis pathway of plants.

MicroRNA profiles and their control of male gametophyte development in rice.

Plant microRNAs (miRNAs) act as negative regulators of gene expression by slicing target transcripts or inhibiting translation. A number of miRNAs play important roles in development. In order to investigate the potential function of miRNAs during male gametogenesis in rice, we obtained both gene and small RNA expression profiles by combining microarray and high-throughput sequencing technologies. From the microarray datasets, 2,925 male gametophyte-specific genes were identified, including 107 transcription factors and three significant Argonaute genes (AGO12, AGO13, and AGO17). From the sRNA-Seq datasets, 104 unique miRNAs (miRus) were identified, including 47 known miRus and 57 novel miRus; interestingly, most of the new miRus are pollen-specific and not conserved among species. Furthermore, an interactive network of miRNA-target was constructed based on the two datasets. By employing enrichment analysis, the miRNA-regulated targets were found to be involved in both the up and down pathways, but predominantly in the down pathways, including 37 GO biological processes and 32 KEGG pathways. These findings indicate that miRNAs play a broad regulatory role during male gametophyte development in rice.

Cloning, overexpression, purification, and characterization of a new iron superoxide dismutase from Jatropha curcas.

A novel iron superoxide dismutase (Fe-SOD) gene from Jatropha curcas was cloned and expressed in Escherichia coli BL21 (DE3). This recombinant enzyme was isolated by a combined procedure involving immobilized metal-ion affinity chromatography and ion-exchange chromatography. The apparent molecular mass of this purified enzyme (designated as JcFe-SOD) was 35 kDa on SDS-PAGE. The full-length complementary DNA sequence of JcFe-SOD contained a 918-bp open reading frame encoding a 305-amino-acid precursor of 34.589 kDa. The result of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry showed that the purified enzyme may own two forms: a dimer and a monomer. The enzyme was relatively stable and showed 54% activity when incubated in 70°C for 60 Min. JcFe-SOD was found to have good pH stability in the pH range of 5.5-9.5 at 25°C over 1 H incubation. The activity of this enzyme was gradually inhibited by increasing concentration of H2O2, 2-mercaptoethanol, and ethylenediaminetetraacetic acid. An assay of the atomic absorption spectrum showed the presence of 0.41 atom of Fe in each SOD subunit.

[Self-microemulsifying drug delivery system increasing solubility and intestinal absorption in situ of tanshinones].

OBJECTIVE: Study the effect of self-microemulsifying drug delivery system(SMEDDS) on the solubility and absorption of tanshinones to guide the selection of composition of tanshinone SMEDDS. METHOD: The solubility of tanshinones in the solution of SMEDDS was determined by UV-spectrometer and the absorption of tanshinone SMEDDS was determined by HPLC as the detection method. RESULT: The solubility of tanshinones in solution of SMEDDS was 10 times in water and 2.5 times in micelle solution. The solubility of tanshinones in solution of SMEDDS was increased with the increasing of oil (MCT) in composition of tanshinone SMEDDS. The absorption constants (Ka) in SMEDDS and micelle solution was 0.479 h(-1) and 0.326 h(-1) respectively, and the absorption half life (t1/2) was 1.44 h and 2.12 h respectively. The absorption was increased with the oil increasing in composition of tanshinone SMEDDS. CONCLUSION: SMEDDS can increase the solubility and absorption of tanshinones significantly and the increasing of oil content (MCT) in SMEDDS composition promote the dissolution and absorption of tanshinones.

Molecular cloning and characterization of phospholipase D from Jatropha curcas.

Phospholipase D (PLD, EC 3.1.4.4) is a key enzyme involved in phospholipid catabolism, initiating a lipolytic cascade in membrane deterioration during senescence and stress, which was cloned from Jatropha curcas L., an important plant species as its seed is the raw material for biodiesels. The cDNA was 2,886 bp in length with a complete open reading frame of 2,427 bp which encoded a polypeptide of 808 amino acids including a putative signal peptide of 53 amino acid residues and a mature protein of 755 amino acids with a predicted molecular mass of 86 kD and a pI of 5.44, having two highly conserved HKD' motifs. Phylogenetic analysis indicated the J. curcas PLD alpha (JcPLDalpha) showed a high similarity to other PLD alpha from plants. Semi-quantitative RT-PCR analysis revealed that it was especially abundant in root, stem, leaf, endosperm and flower, weakly in seed. And the JcPLDalpha was increasedly expressed in leaf undergoing environmental stress such as salt (300 mM NaCl), drought (30% PEG), cold (4degreeC) and heat (50degreeC). The JcPLDalpha protein was successfully expressed in Escherichia coli and showed high enzymatic activities. Maximal activity was at pH 8 and 60degreeC.

[Design, optimization and quality evaluation of curcumin self-emulsifying drug delivery system (SEDDS)].

OBJECTIVE: To develop the self-emulsifying drug delivery system (SEDDS) of curcumin, and evaluate its quality in vitro. METHODS: The excipients of curcumin SEDDS were selected via its solubility study in various oils, surfactants and co-surfactants and the formulation was optimized using ternary phase diagram study and central composite design-response surface methodology (oil, surfactant and co-surfactant percentages as factors; solubility, droplet size, polydispersity index and emulsifying time as responses). The appearance, droplet size and polydispersity index after emulsifying and the emulsifying time of optimized curcumin SEDDS were studied. The solubility of curcumin in the solution of SEDDS was determined. RESULTS: Castor oil-(tween-80) -ethanol = 28: 55: 20 (w/ w/w) was selected for optimum curcumin SEDDS. The droplet size was 222. 2 nm, polydispersity index was 0. 171. The time of self-emulsifying was 10 s and the solubility of curcumin in SEDDS was 1.93 mg/mL. CONCLUSION: Curcumin SEDDS formulation is selected and optimized successfully, and the preparation of curcumin SEDDS is simple, the quality is stable.

[Clinical effect and mechanismn of xuefu zhuyu capsule in treating unstable angina pectoris].

OBJECTIVE: To observe the clinical effect of Xuefu Zhuyu Capsule (XFZYC) in treating unstable angina pectoris (UAP) and to investigate its mechanism of protection on vascular endothelia. METHODS: Sixty UAP patients were randomly assigned to two groups, the 30 in the treated group were treated by conventional therapy plus XFZYC, and the 30 in the control group treated by conventional therapy alone. The frequency and persistent time of angina pectois, dosage of nitroglycerin used, changes of electrocardiogram (ECG) were observed, and the plasma levels of endothelin (ET), nitric oxide (NO), von Willebrand factor (vWF), soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1) were tested before and after the two-month therapeutic course. RESULTS: (1) The clinical symptoms as frequency and persistent time of angina pectoris in the treated group were bettered significantly after treatment and the dosage of nitroglycerin used decreased (all P < 0.01), showing significant difference when compared with those in the control group (P < 0.05). (2) The effective rates on UAP symptoms and ECG in the treated group were 86.7% and 76.7% respectively, which were significantly higher than those in the control group (70.0% and 63.3%, P < 0.05). (3) Laboratory examination showed in the control group, changes only displayed in the decrease of vWF and ET and increase of NO (all P < 0.05), while in the treated group, plasma levels of vWF, ET, sVCAM-1 and sICAM-1 after treatment significantly decreased and were lower than those in the control group (P < 0.01), but NO elevated significantly and was higher than that in the control group (P < 0.01). CONCLUSIONS: XFZYC is an effective Chinese patent medicine for treatment of UAP. It displays its effect on protecting vascular endothelia and anti-angina pectoris partially by decreasing the plasma levels of ET, vWF, sVCAM-1 and sICAM-1 and elevating the level of NO.

[Analysis on tongue manifestations of 488 cases of Warm disease in Qing Dynasty].

488 cases of warm disease with tongue diagnosis picked up from 29 monographs providing materials of tongue diagnosis are sorted out of 51 monographs of medical records in the Qing Dynasty. Statistics and analysis were made on these 488 cases of tongue diagnoses, according to the classification of tongue manifestations in the Textbook of Diagnostics of Chinese Medicine for TCM schools of higher education. Results showed that, among all the cases, the recording rate of tongue fur was 65. 16% and was the highest one of the recording materials. The frequencies of different fur recorded from high to low were yellow, white and black. From 1850 to 1911 in the Qing Dynasty, occurrence rate of tongue coating had changed. Those of white, yellow and compound fur were increased. On the contrary, that of black fur was decreased. The recording rate of the color of tongue proper was 23. 16%, among which the proportions of red and crimson tongue were the highest. The recording rate of fur quality of fur was 37. 7%. From 1850 to 1911 in the Qing Dynasty, the recording rate of greasy fur reached the first place in fur quality. Tongue diagnosis was applied in warm- heat and damp-heat types of Warm Disease and pestilence.