Dual-function DEFENSIN 8 mediates phloem cadmium unloading and accumulation in rice grains.

Grain cadmium (Cd) is translocated from source to sink tissues exclusively via phloem, though the phloem Cd unloading transporter has not been identified yet. Here, we isolated and functionally characterized a defensin-like gene DEFENSIN 8 (DEF8) highly expressed in rice (Oryza sativa) grains and induced by Cd exposure in seedling roots. Histochemical analysis and subcellular localization detected DEF8 expression preferentially in pericycle cells and phloem of seedling roots, as well as in phloem of grain vasculatures. Further analysis demonstrated that DEF8 is secreted into extracellular spaces possibly by vesicle trafficking. DEF8 bound to Cd in vitro, and Cd efflux from protoplasts as well as loading into xylem vessels decreased in the def8 mutant seedlings compared with the wild type. At maturity, significantly less Cd accumulation was observed in the mutant grains. These results suggest that DEF8 is a dual function protein that facilitates Cd loading into xylem and unloading from phloem, thus mediating Cd translocation from roots to shoots and further allocation to grains, representing a phloem Cd unloading regulator. Moreover, essential mineral nutrient accumulation as well as important agronomic traits were not affected in the def8 mutants, suggesting DEF8 is an ideal target for breeding low grain Cd rice.

Luminescent Pt(II) Complexes Containing (1-Aza-15-crown-5)dithiocarbamate and (1-Aza-18-crown-6)dithiocarbamate: Mechanochromic and Solvent-Induced Luminescence.

The strong tendency to stack in the solid state and rich luminescence for the Pt(II) complexes makes them potential candidates as new mechanochromic materials and sensing applications. Six mononuclear complexes [Pt(ppy)(O4NCS2)] (1), [Pt(bpy)(O4NCS2)]ClO4 (2), [Pt(ppy)(O5NCS2)] (3), [Pt(phen)(O4NCS2)]ClO4·CH3OH (5a), [Pt(phen)(O4NCS2)]ClO4 (5b), and [Pt(phen)(O5NCS2)]ClO4 (6a), one dinuclear complex [Pt2(phen)2(NaO5NCS2)2(ClO4)3]ClO4 (6b), and one one-dimensional (1-D) coordination polymer {[Pt2(bpy)2(NaO5NCS2)2(ClO4)2](ClO4)2}n (4) were synthesized by reacting [Pt(ppy)Cl]2, Pt(bpy)Cl2, and Pt(phen)Cl2 (ppy = 2-phenylpyridine, bpy = 2,2'-bipyridine, and phen = 1,10-phenanthroline) with (1-aza-15-crown-5)dithiocarbamate (O4NCS2) or (1-aza-18-crown-6)dithiocarbamate (O5NCS2), respectively, which have been isolated and structurally characterized by X-ray diffraction. Neutral complexes 1 and 3 contain no intermolecular Pt(II)···Pt(II) contact, whereas cationic complexes 2, 5a, 5b, and 6a with ClO4- as counteranions show alternative intermolecular Pt(II)···Pt(II) contacts of 3.535/4.091, 3.480/5.001, 3.527/4.571, and 3.446/4.987 Å in the solid state, respectively. Interestingly, complex 4 forms a 1-D coordination polymer through coordination between the encapsulated Na+ ions inside the azacrown ether rings of O5NCS2 and ClO4- anions with respective intra- and intermolecular Pt(II)···Pt(II) contacts of 3.402 and 3.847 Å in crystal lattices, whereas a dinuclear complex 6b was surprisingly formed and also connected by the encapsulated Na+ ions and ClO4- anions with alternative intra- and intermolecular Pt(II)···Pt(II) contacts of 3.650 and 3.677/4.4.372 Å, respectively. Upon excitation, complexes 1 and 3 showed similar vibronic luminescence at 507, 534, and 502, 532 nm, respectively, and the other complexes 2 and 4-6 showed broad luminescence with maxima at 537-567 nm. The B3LYP/LanL2DZ calculation was carried out and used to clarify their excited-state properties. In addition, the powder samples for complexes 1-4 almost showed no energy shift for the luminescence and significantly those of complexes 5-6 exhibited the mechanochromic luminescence upon grinding. It is noted that complexes 5a and 6a only showed minor red shifts (i.e., from 544 to 556 nm for complex 5a and from 551 to 565 nm for complex 6a), whereas complex 6b exhibited a remarkable red shift from 558 to 603 nm upon grinding. Besides, their luminescence reversibility was also examined toward various solvents.

[Content determination of seven active components of Eucommiae Cortex in aortic vascular endothelial cells of spontaneously hypertensive rats by UPLC-MS/MS].

In the present study, the contents of seven active components [genipinic acid(GA), protocatechuic acid(PCA), neochlorogenic acid(NCA), chlorogenic acid(CA), cryptochlorogenic acid(CCA),(+)-pinoresinol di-O-ß-D-glucopyranosid(PDG), and(+)-pinoresinol 4'-O-ß-D-glucopyranoside(PG)] of Eucommiae Cortex in aortic vascular endothelial cells of spontaneously hypertensive rats(SHR) were simultaneously determined by ultra-high liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS). The qualified SHR models were selected. The primary aortic endothelial cells(VECs) of rats were separated and cultured by ligation and adherence, followed by subculture. After successful identification, an UPLC-MS/MS method for simultaneously determining the contents of GA, PCA, NCA, CA, CCA, PDG, PG in seven components of Eucommiae Cortex in VECs was established, including specificity, linearity, matrix effect, recovery, accuracy, precision and stability. The established method had the lo-west limit of quantification of 0.97-4.95 µg·L~(-1), accuracy of 87.26%-109.6%, extraction recovery of 89.23%-105.3%, matrix effect of 85.86%-106.2%, and stability of 86.00%-112.5%. Therefore, the established accurate UPLC-MS/MS method could rapidly and simultaneously determine the contents of the seven active components of Eucommiae Cortex in VECs of SHRs, which provided a refe-rence for the study of cellular pharmacokinetics of active components of Eucommiae Cortex extract.

[Establishment of PK-PD model in anti-inflammatory active components in Inula cappa extract based on lipopolysaccharide-induced in vitro inflammation model].

In the present study, a pharmacokinetics(PK)-pharmacodynamics(PD) model in the anti-inflammatory active components in Inula cappa extract was established based on the lipopolysaccharide(LPS)-induced in vitro inflammation model in order to clarify the relationship between the dynamic changes of anti-inflammatory active components in inflammatory cells and their efficacy. Firstly, the inflammation model in vitro was induced by 1 µg·mL~(-1) LPS in RAW264.7 cells for 24 h. After treatment with 400 µg·mL~(-1) I. cappa extract, the pharmacokinetics(PK) of five anti-inflammatory active components, including luteolin(LUT), chlorogenic acid(CA), cryptochlorogenic acid(CCA), 3,4-dicaffeoylquinic acid(3,4-DCQA), and 4,5-dicaffeoylquinic acid(4,5-DCQA), in normal cells and inflammatory cells was compared. Meanwhile, the PD study was carried out by measuring the inflammatory factors NO and TNF-α in the cell supernatant at each time point, which was fitted with PK by the Phoenix Model in the WinNonlin 8.2 to establish the PK-PD model for five components including LUT, CA, CCA, 3,4-DCQA, and 4,5-DCQA. The results showed that compared with normal cells, the model cells showed increased or decreased uptake of five components, advanced T_(max), faster absorption, prolonged MRT and t_(1/2), and increasing or decreasing trend of CL_(z/F) and V_(z/F). When NO was used as the efficacy index, the PK-PD model after the integration of the multi-effect components in I. cappa was E=7.45×\[1-Ce~(5.74)/(78.24~(5.74)+Ce~(5.74))\], while with TNF-α as the efficacy index, the PK-PD model after the integration of the multi-effect components in I. cappa was E=79.28×[1-Ce~(6.45)/(85.10~(6.45)+Ce~(6.45))]. The results of the study suggested that the inflammatory state could change the cellular PK of I. cappa. The anti-inflammatory effect of active components in I. cappa might be related to the down-regulation of the secretion of NO and TNF-α in inflammatory cells, and NO and TNF-α might serve as the anti-inflammatory targets of active components of I. cappa.

[Differences in intestinal absorption characteristics of Cynanchum auriculatum extract based on in situ intestinal circulation perfusion model in two states].

The present study aimed to investigate the intestinal absorption characteristics of six components(syringic acid, scopoletin, baishouwu benzophenone, caudatin, qingyangshengenin, and deacylmetaplexigenin) in Cynanchum auriculatum extract. In situ intestinal circulation perfusion model was employed to investigate the differences in intestinal absorption characteristics of C. auriculatum extract under the influence of different intestinal segments, different drug concentrations, and bile in the normal and functional dyspepsia(FD) states. The results showed that the absorption of baishouwu benzophenone decreased with the increase in the concentration of C. auriculatum extract(P<0.01), while the absorption of syringic acid and other components increased in a dose-independent manner, suggesting that baishouwu benzophenone might follow active absorption, while other components might not be on a single absorption pattern. The main absorption sites of each component in the normal state were different from those in the FD state. The cumulative absorption conversion rates in the FD state were generally lower than those in the normal state, and bile inhibited the absorption of other components except for scopoletin in both states(P<0.05). As revealed, the small intestine showed selectivity to the absorption of drugs, and the pathological state(such as FD) and bile both affected the absorption of the main components, which provides a theoretical basis for the development of new drugs and further development of C. auriculatum.

[Uptake and transport of Laportea bulbifera extract in Caco-2 cell model].

Laportea bulbifera extract is effective in resisting inflammation and shows a good therapeutic effect on rheumatoid arthritis in rats. However, the absorption characteristics of active components in L. bulbifera extract in Caco-2 cells are still unclear, which limits the in-depth development of L. bulbifera resources. The purpose of this study was to investigate the absorption and transport mechanism of the active components of L. bulbifera extract in the Caco-2 cell model and explore the effects of different factors(concentration, time, pH value, temperature, and efflux transporter inhibitor) on its uptake and transport. The results showed that L. bulbifera extract at the concentration of 2.0-8.0 mg·mL~(-1) showed no toxicity to Caco-2 cells. The uptake and transport of L. bulbifera extract in the Caco-2 cell model were concentration-dependent and time-dependent. The main absorption mechanism was passive diffusion, and acidic condition(pH 5.0-6.0) and 37 ℃ were more favorable for drug absorption. P_(app)>1.0×10~(-6 )cm·s~(-1) of each component indicated that L. bulbifera was a moderately absorbed drug. P-gp, MRP2, and BCRP were not involved in its uptake and transport.

[Potential pharmacodynamic substances of Laportea bulbifera in treatment of rheumatoid arthritis based on serum pharmacochemistry and pharmacology].

The present study investigated the pharmacodynamic material basis of Laportea bulbifera in the treatment of rheumatoid arthritis. Firstly, human rheumatoid arthritis fibroblast-like synoviocyte line MH7A was cultured in vitro and treated with tumor necrosis factor alpha(TNF-α, 50 ng·mL~(-1)). The proliferation and the levels of inflammatory cytokines such as prostaglandin E2(PGE2), interleukin-1ß(IL-1ß), and interleukin-6(IL-6) of the MH7A cells exposed to the serum containing L. bulbifera were determined to evaluate the anti-rheumatoid arthritis effects of the serum. Furthermore, the ultra-performance liquid chromatography tandem mass spectrometry fingerprints of the L. bulbifera crude extract, the drug-containing serum, and the drug-free serum were compared to identify the compounds newly generated in the serum after oral administration of the extract. According to the peak areas of common peaks and the results of anti-rheumatoid arthritis effect test, the active components were identified. The serum containing L. bulbifera significantly inhibited the proliferation of the MH7A cells activated by TNF-α and the expression of PGE2, IL-6, and IL-1ß. Thirty newly generated compounds were detected in the drug-containing serum. Among them, neochlorogenic acid, cryptochlorogenic acid, chlorogenic acid, rutin, isoquercitrin, luteoloside, kaempferol-3-O-rutinoside, and quercitrin were also present in the crude extract. Twelve characteristic peaks(3, 7, 8, 14, 18, 19, 21, 23, 24, m6, m7, and m15) were significantly correlated with the pharmaceutical effect. According to the correlations, neochlorogenic acid, cryptochlorogenic acid, and chlorogenic acid had great contributions to the anti-rheumatoid arthritis activity. This study preliminarily clarified the potential pharmacodynamic substances of L. bulbifera in the treatment of rheumatoid arthritis, which laid a theoretical and experimental foundation for further development and application of the medicinal plant.

[Effect of Laportea bulbifera extract on CYP450 enzyme activities in rats by Cocktail probe drug method].

The Cocktail probe drug method was used to evaluate the effect of Laportea bulbifera extract on the different subtypes of CYP450 enzyme activities in rats, and to provide references for its clinical rational drug use. The rats were randomly divided into a high-dose L. bulbifera group(0.45 g·kg~(-1)·d~(-1)) and a low-dose L. bulbifera group(0.09 g·kg~(-1)·d~(-1)). After continuous gavage for 7 and 14 days, the Cocktail probe mixing solution, including caffeine, midazolam, tolbutamide, omeprazole, metoprolol, and chlorzoxazone, was injected into the tail vein, and the blood sample was obtained from the tail vein at different time points. Ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS) was established to determine the concentration of six probe drugs in rat plasma. DAS 2.0 was used to calculate its pharmacokinetic parameters, and the effect of L. bulbifera extract on CYP1 A2, CYP2 C9, CYP2 C19, CYP2 D6, CYP2 E1, and CYP3 A4 in rats was investigated. As compared with the blank control group, under the omeprazole index, the AUC_(0-t) and AUC_(0-∞) of the 7-day administration groups and the 14-day high-dose group were significantly decreased, and the CLz and Vz were significantly increased. Under the midazolam index, the AUC_(0-t) and AUC_(0-∞) of the 7-day low-dose group and the 14-day administration group were significantly decreased, and the CLz was significantly increased. In addition, the AUC_(0-∞) of the 7-day high-dose group was also significantly decreased. Under the index of metoprolol, the AUC_(0-t) and AUC_(0-∞) of each experimental group were decreased significantly, and the CLz and Vz of the 7-day low-dose group and the 14-day low-dose group were increased significantly. Under the caffeine index, the AUC_(0-t) and AUC_(0-∞) of the 7-day administration groups were increased significantly, the CLz was decreased significantly, and the t_(1/2 z) of the 14-day high-dose group was increased significantly. Under the chlorzoxazone index, the AUC_(0-t) and AUC_(0-∞) of the 7-day low-dose group were increased significantly, and the CLz was decreased significantly. Under the tolbutamide index, there was no statistical difference in the pharmacokinetic parameters. In conclusion, L. bulbifera extract induces the activities of CYP2 C19, CYP3 A4, and CYP2 D6, inhibits the activities of CYP1 A2 and CYP2 E1, and does not affect the activity of CYP2 C9.

Solvent-Induced Luminescence and Structural Transformation of a Dinuclear Gold(I) (Aza-18-crown-6)dithiocarbamate Compound.

The reaction of AuCl(SMe2) with equimolar NaO5NCS2 [O5NCS2 = (aza-18-crown-6)dithiocarbamate] in CH3CN gave [Au2(O5NCS2)2]·2CH3CN (2·2CH3CN), where six other 2·solvates (solvates = 2DMF, 2DMSO, 2THF, 2acetone, 1.5toluene, and 1.5anisole) can be successfully isolated from different crystal-growing processes (i.e., ether diffusion, layer method, or evaporation in air) by dissolving the dry powder samples of 2·2CH3CN in the respective solvents, and their crystal structures are all determined by X-ray diffraction as well. It is noted that there are different intermolecular Au(I)···Au(I) contacts in combination with various luminescences for 2·solvates and indeed there is a close relationship between intermolecular Au(I)···Au(I) contacts [i.e., 2.8254(7)-2.9420(5) Å] and luminescence energies (i.e., 554-604 nm), including three examples of 2·2CH3CN, 2·0.5m-xylene, and 2·tert-butylbenzene·H2O reported in our previous work. In 2·solvates, the toluene and tert-butylbenzene solvates have the shortest [2.8254(7)-2.8289(7) Å] and longest [2.9420(5) Å] intermolecular Au(I)···Au(I) contacts, respectively, and consequently they show the respective lowest (604 nm) and highest (554 nm) luminescence energies. Indeed, 2·solvates exhibit different types of time-dependent luminescence upon solvate loss in air. Furthermore, B3LYP/LanL2DZ calculation results can help to clarify the relationship between intermolecular Au(I)···Au(I) contacts and luminescence energies for 2·solvates.

First Report of Fruit Rot of Grapes (Vitis vinifera) Caused by Cladosporium cladosporioides in Xinjiang, China.

Xinjiang Province accounts for nearly 20% of the total grape (Vitis Vinifera L.), proles orientalis) (wine, table and raisin combined) production, being the largest production area in China. Fruit rot is the most common disease that impacts grape quality and yield. A new disease where the ripe grape berry surfaces were coated with brownish mildew was observed, and the disease mainly occurred on whole clusters or most of the berries in the cluster. In September 2019 and 2020, 125 diseased grape clusters were collected from 10 locations in northern Xinjiang where the disease incidence was 15.3% - 27.4% ((diseased clusters/ total clusters)*100). To identify the pathogen, symptomatic grape berries were disinfected with 1% NaClO for 2 min, followed by 70% ethanol for 30 s, and rinsed thrice in sterile distilled water. Three pieces of ~0.5 cm2 diseased grape skin with partial exocarp were placed on potato dextrose agar (PDA) amended with streptomycin sulfate and kanamycin (50 µg/mL each). The PDA plates were then incubated at 25℃ under light condition with the luminous intensity 3500 Lux for 7 days. Fungal colonies emerging from the plated tissue were subcultured and single-spored three times to obtain pure cultures. From 20 strains with similar colony phenotype and grey olive hue, flocculent, felt-like surface, six (Cc-Vivi-3, 7, 9, 11, 13 and 19) isolates were chosen for further characterization after 7 days of incubation. Conidia were either single or grew in chains, with around 4 conidia per chain. Conidia were ovoid, nearly spindle or globose with slightly smooth or irregular reticulate surface. Conidiophores were solitary, smooth, septate, erect or geniculate. These characteristics were consistent with the descriptions for Cladosporium cladosporioides. To confirm this identification, PCR was performed on the genomic DNA of the selected strains using primers for internal transcribed spacer (ITS) region ITS1/ITS4, actin (ACT) and translation elongation factor (TEF) (Schubert et al., 2007; Braun et al., 2003). Amplified ITS sequences provided a 100% match to C. cladosporioides (AY213641) in NCBI. Homology of ACT sequences to C. cladosporioides (HM148527 and MH047330) was 99.57% and 100%, respectively; and the homology of TEF sequences with C. cladosporioides (HM148258, HM148289, HM148260 and HM148266) was 97.56% ~ 100%. To further confirm the evolutionary relationship of strains from grapes with Cladosporium spp., phylogenetic analyses based on ITS, ACT and TEF conjoint sequences from the six experimental isolates, five C. cladosporioides strains, eight proximal Cladosporium species were analyzed. The phylogenetic tree showed that the six isolates from grapes clustered with C. cladosporioides strains, but not other proximal Cladosporium species. This confirmed that all six isolates evaluated were C. cladosporioides. Pathogenicity tests with one C. cladosporioides isolate (Cc-Vivi-3; accession No. ITS: MW556429, ACT: MW567144, TEF: MW567143) were carried out as follows: ripe and healthy grape clusters from cultivars Xinyu and Munag when total soluble solids were 20-21°Bx and 19-20 °Bx, respectively, were detached from the vines. Five berries of three clusters of each cultivar were punctured with a sterile syringe, then inoculated with a 20 µL conidial suspension (107 conidia/mL). And uninoculated, punctured berries in clusters treated with sterilized water served as controls. The experiment was repeated three times. Symptoms were recorded 15 days after incubation at 80% relative humidity and 25℃ with a 14 h light/10 h dark cycle. The olive green or blackish green mildew layer was produced on all inoculated berries. No symptoms were observed on the uninoculated berries. Koch's postulates were fulfilled by reisolating C. cladosporioides from all symptomatic tissues and identifying them by PCR targeting the ACT gene. This is the first description of C. cladosporioides causing grape fruit rot in Xinjiang, China. In recent years, worldwide reports of Cladosporium spp. damaging crops are increasing (Briceño et al., 2008; Walker et al., 2016; Meneses et al., 2018; Robles-Yerena et al., 2019; Ding et al., 2019; Yang et al., 2021). However, relatively few methods of management including some fungicides and biocontrol agents are available in different crops (Wang et al., 2018; Addrah et al., 2019). In view of the important role of Xinjiang in China agricultural production, that should arouse strong attention.

Association between magnetic resonance imaging of carotid artery and coronary stenosis detected by computed tomography angiography.

OBJECTIVE: To investigate the correlation between carotid artery stenosis (CAS) measured by magnetic resonance imaging (MRI) and the coronary stenosis (CS) determined by computed tomography angiography (CTA). METHODS: In this prospective study, 42 subjects diagnosed with coronary artery disease (CAD) underwent MRI and CTA examinations. The severity degree and number of CAS, the score, detection rate and type of carotid plaque, and also the severity degree and number of CS were assessed. Spearman's rank correlation test was used to evaluate the correlation between CAS and CS. RESULTS: CS was detected in 42 (100%) subjects, while CAS was detected in 36 (85.7%) subjects. Distribution of CAS severity grades in multiple-vessel group was significantly different from other groups, which with more moderate and severe stenosis (p < 0.05). A positive and significant correlation between the CAS severity and CS severity (r = 0.612, p < 0.05), and the number of involved coronary vessels (r = 0.572, p < 0.05) were observed, respectively. Both detection rate (r = 0.587, p < 0.05) and score (r = 0.735, p < 0.05) of carotid plaque showed a good correlation with the number of involved coronary vessels. After carotid MRI, 71 carotid plaques were detected in 42 subjects, with an incidence rate of 9.9% in subjects with mild CS, 18.3% in moderate CS and 71.8% in severe CS. CONCLUSION: Correlation between CAS measured by MRI and CS determined by CTA was identified in present study. These results indicated that the non-invasive CAS evaluation employing the MRI may be clinically useful for the assessment of CS.

[Study on metabolites of Laportea bulbifera extract in rat feces based on UHPLC-Q-TOF-MS~E technique].

This project is to study the metabolites of Laportea bulbifera extract in rat feces. After the SD rats were gavaged with the extract(136 g·kg~(-1), according to the crude drug dose), the metabolites in their feces were detected by UHPLC-Q-TOF-MS~E technique, and the obtained mass spectrometry data was combined with UNIFI software for prediction. The prototype components and metabolites in rat feces were identified with reference materials and related literature. A total of 43 metabolites were identified(including 8 prototype components and 35 metabolites). The metabolic pathways mainly include monocaffeoylquinic acid(hydrogenation reduction, ring-opening cracking, sulfation, hydroxylation, glucuronidation), quercetin(O-C2 bond ring-opening cleavage, C2-C3 double bond reduction, rutin carbonylation) and so on. The metabolites and metabolic process of L. bulbifera extract in rat feces were clarified, which provided a basis for the study of the active substances and its mechanism of action.

[Intestinal absorption properties of Gastrodiae Rhizoma powder with different particle size by circulation pass perfusion model in rats].

This study is to study the absorption properties of different particle size of Gastrodiae Rhizoma powder in rats. In vivo circulation pass perfusion model combined with ultra high performance liquid chromatography-mass spectrometry(UPLC-MS/MS) method was used to determine the cumulative absorption of each component in different particle size of Gastrodiae Rhizoma powder, and the effect of different particle size, different concentrations, different intestine segments and bile on the intestine absorption of gastrodin and other compositions in Gastrodiae Rhizoma powder was investigated to illuminate the absorption properties and compare the absorption difference of gastrodin and other compositions in Gastrodiae Rhizoma powder in different particle size. The results showed that the absorption of gastrodin in each intestinal segment has no significant difference, pointing out that gastrodin may be passive absorption and the absorption of barrison glycosides may be active absorption; the absorption of gastrodin in ultrafine powder was better than that of common powder and superfine powder of Gastrodiae Rhizoma; the absorption of these barrison glycosides was good in ultrafine powder of Gastrodiae Rhizoma under the high concentration. However, an appropriate degree of superfine grinding can promote the absorption of active ingredients of Gastrodiae Rhizoma. This test can provide information for the deep development of Gastrodiae Rhizoma.

[Differences in intestinal absorption characteristics of Laportea bulbifera extract in normal and rheumatoid arthritis model rats by isolated everted intestine model].

This work aimed to investigate the intestinal absorption characteristics of Laportea bulbifera extract in normal and rheumatoid arthritis model rats. The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, rutin, kaempferol-3-O-rutinoside, galuteolin, quercetin and isoquercetin in intestinal absorption solution samples were detected by UPLC-MS/MS with 5.0 g·L~(-1) as the absorption concentration. The cumulative absorption(Q) and absorption rate constant(K_a) were calculated, and the absorption characteristics of different components of L. bulbifera in intestinal absorption solution of normal rats and rheumatoid arthritis rats were compared. The results showed that all the eight index components in the extract of L. bulbifera could be absorbed into the intestinal capsule, the cumulative absorption-time curve of each component showed an upward trend without saturation, and the correlation regression coefficient(R~2) was greater than 0.92, which is consistent with the zero-order absorption rate process. It was speculated that the possible absorption mode of each component was passive diffusion. In normal condition, the absorption of ileum was the best(except chlorogenic acid), and in pathological condition, duodenum was the best. The total absorption of 8 components in each intestinal segment of RA rats was better than that of normal rats, which speculated that rheumatoid arthritis may change the specific site of drug absorption. The experimental results showed that rheumatoid arthritis could change the intestinal absorption of the extract of L. bulbifera, and its mechanism needs further study.

Glucocorticoid receptor in ayu (Plecoglossus altivelis): Genomic and non-genomic effects on monocytes/macrophages function.

The glucocorticoid receptor (GR) is an important feedback regulator of the hypothalamic-pituitary-interrenal (HPI) axis. However, there are a limited number of studies focused on host-pathogen interactions in which an association between GR and immune response has been evaluated in monocytes/macrophages (MO/MФ) after being challenged with highly pathogenic bacteria. Here, we cloned the cDNA sequence of the glucocorticoid receptor (PaGR) gene from ayu fish. The PaGR transcript was expressed in all tissues, and changes in expression were observed in immune tissues and MO/MФ after live Vibrio anguillarum infection. Subsequently, PaGR was expressed and purified to prepare anti-PaGR antibodies. We analyzed the subcellular localization of PaGR. PaGR was expressed not only in the intracellular space but also in the plasma membrane. PaGR activation decreased the expression of pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokines. However, PaGR activation suppressed the phagocytosis activity of V. anguillarum-infected ayu MO/MФ via a non-genomic pathway. Interestingly, PaGR activation could enhance MO/MФ bacterial killing capability and apoptosis. Therefore, PaGR may modulate the immune response in ayu MO/MФ by genomic and non-genomic pathways.

[UPLC-Q-TOF-MS/MS analysis on prototypes components and metabolites of effective fractions of Polygonum orientale flower in rat serum and urine].

Ultra performance liquid chromatography coupled with time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) method was applied to analyze the prototypes and metabolites of the effective components of Polygonum orientale in SD rat serum and urine. The separation was performed on Agilent Eclipse Plus C_(18) column( 2. 1 mm×100 mm,1. 8 µm),with 0. 1% formic acid solution( A)-acetonitrile( B) as the mobile phase for gradient elution. Mass spectrometry data of biological samples were obtained under positive and negative electrospray ion mode. By comparing chromatogram differences between blank samples and drug treatment samples,prototype components and metabolites of the effective components of P. orientale extract were identified. The results showed that 12 metabolites were detected in serum and 26 metabolites in urine( including cross-components) of rats. The main metabolic pathways included hydrogenation,hydroxylation,glucuronidation,sulfation reaction,and methylation-glucuronidation,etc. The method established in this study was reliable and effective for studying the metabolic characteristics of the effective components of P. orientale in rats,and it can provide a reference for further studies on therapeutic material basis of this herb.

[Absorption of Inula cappa extract based on everted intestinal sac method].

To investigate the absorptive characteristics of Inula cappa extract based on the rat everted intestinal sac method in vitro. Nine representative ingredients in I. cappa extract were selected as the study objects. An UPLC-MS/MS method was established to determine and detect their cumulative absorption amount for expounding the absorptive characteristics of ingredients in different intestinal sections. According to the results， the transport mechanism of 8 compounds showed passive diffusion by the reverted gut sac method. And scopolin was actively transported in the intestine. The best absorption site of chlorogenic acid was duodenum. The best absorption site of cryptochlorogenic acid， 1，3-O-dicaffeoylquinic acid， luteolin-7-glucoside and 3，4-O-dicaffeoylquinic acid were jejunum. The best absorption site of neochlorogenic acid， scopolin， 4，5-O-dicaffeoylquinic acid and 3，5-O-dicaffeoylquinic acid was ileum. The absorption of all the compounds was affected by pH and bile. All of the nine ingredients in I. cappa extract could be absorbed in intestines， but with differences in the absorption rate， the best absorptive site and mechanism， indicating that the intestinal absorption of I. cappa extract was selective.

The Improved Suppression and Analysis of Stray Light in the Miniature Raman Spectrometer.

Stray light analysis has great benefits in designing Raman spectrometry system, which is the sensitive detection system of weak optical signal. In this paper, optical design software and Solidworks are utilized to optimize the optical and mechanical structure. The system resolution is 0.7 nm, and the volume is 110 mm×95 mm, which belongs to portable and miniature Raman spectrometer. Based on the stray light simulation model, we made an analysis of ray tracing simulation for this system. First, the stray light come with the incident ray were suppressed by the aperture stop. Then the receiver of stray light was introduced and improved in the design progress to suppress internal stray light, especially for zero-order diffraction light of plane grating. The improved receiver of stray light is more effectively using the internal space of the spectrometer and analysis results show that a 50% reduction in the number of stray light and stray light normalized irradiance intensity from 10-5 down to 10-7. The analysis shows that the improved receiver of stray light can effectively suppress stray light, which is beneficial to weak signal detection, and provide reference for design and adjustment of the miniature Raman spectrometer.

[Simultaneous Determination of Protocatechuic Acid,Kaempferol Biotin Glucoside and Quercitrin in Rat Plasma and Pharmacokinetics By UPLC-MS/MS].

Objective: To develop a sensitive and reliable ultra performance liquid chromatography tandem mass spectrometry ( UPLC-MS / MS) method for simultaneous determination and pharmacokinetics of protocatechuic acid, kaempferol biotin glucoside and quercitrin in rat plasma, and study their pharmacokinetic characteristics in rats. Methods: Three compounds were simultaneously determined by UPLC-MS / MS on Waters BEH C18( 50 mm × 2. 1 mm,1. 7 µm) column. The mobile phase was 0. 1% acetonitrile formic acid and0. 1% aqueous formic acid, and programmed in a linear gradient mode. The compounds were ionized in the electrospray ionization ion source of the mass spectrometer and detected in MRM mode. Results: The t1 /2of protocatechuic acid,kaempferol biotin glucoside and quercitrin after intravenous injection were( 41. 9 ± 12. 3),( 71. 3 ± 56. 8) and( 90. 3 ± 74. 8) min. The Cmaxwere( 1. 7 ± 0. 7) µg / m L,( 2. 1 ± 0. 9) µg/m L,( 8. 7 ± 3. 7) µg/m L. Conclusion: The established method is specific,rapid,accurate and sensitive,and is proved to meet the requirements of biological sample analyses,and is suitable for the concentration determination of protocatechuic acid, kaempferol biotin glucoside and quercitrin in rat plasma, three compounds are all best fitted to the two-compartment open pharmacokinetic model.

Directed gas-phase formation of the ethynylsulfidoboron molecule.

As a member of the organo sulfidoboron (RBS) family, the hitherto elusive ethynylsulfidoboron molecule (HCCBS) has been formed via the bimolecular reaction of the boron monosulfide radical (BS) with acetylene (C2H2) under single collision conditions in the gas phase, exploiting the crossed molecular beams technique. The reaction mechanism follows indirect dynamics via a barrierless addition of the boron monosulfide radical with its boron atom to the carbon atom of the acetylene molecule, leading to the trans-HCCHBS intermediate. As predicted by ab initio electronic structure calculations, the initial collision complex either isomerizes to its cis-form or undergoes a hydrogen atom migration to form H2CCBS. The cis-HCCHBS intermediate either isomerizes via hydrogen atom shift from the carbon to the boron atom, leading to the HCCBHS isomer, or decomposes to ethynylsulfidoboron (HCCBS). Both H2CCBS and HCCBHS intermediates were predicted to fragment to ethynylsulfidoboron via atomic hydrogen losses. Statistical (RRKM) calculations report yields to form the ethynylsulfidoboron molecule from cis-HCCHBS, H2CCBS, and HCCBHS to be 21%, 7%, and 72%, respectively, under current experimental conditions. Our findings open up an unconventional path to access the previously obscure class of organo sulfidoboron molecules, which are difficult to access through "classical" formation.