Volatile-compound fingerprinting and discrimination of positional isomers in stamp-pad ink tracing using HS-GC-IMS combined with multivariate statistical analysis.

The rising crime rate associated with document forgery has a significant impact on public safety and social stability. In document fraud cases, determining the origin of a particular stamp-pad ink is the most important objective. In this study, a comprehensive analysis of the volatile compounds in quick-drying stamp-pad inks from six commonly used brands were performed for the first time, utilizing a combination of headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) and multivariate statistical analysis methods. Visual and comparative analysis of the differential volatile components among different stamp-pad ink samples was conducted using fingerprints and volcano plots. A total of 127 volatile compounds were accurately identified, with ketones, esters, alcohols, and aldehydes being the most abundant compounds in the stamp-pad inks. Hierarchical clustering analysis (HCA), including dendrograms and clustering heatmaps, was utilized to explore the correlations between these compounds and the samples. Additionally, the precise identification of positional isomers and functional group isomers of aliphatic compounds was achieved. To achieve accurate discrimination of various stamp-pad ink samples, a multivariate statistical analysis method was utilized to establish a classification model for them. Based on the results obtained from HS-GC-IMS, effective discrimination among different brands of stamp-pad ink samples was achieved through principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The model exhibited excellent performance, with the fit index of dependent variables (R2Y) and the predictive index of the model (Q2) values of 0.99 and 0.984, respectively. These results provided significant theoretical evidence for the application of HS-GC-IMS as an efficient technique in the analysis of volatile compounds, identification of positional isomers and functional group isomers, as well as tracing the origin of stamp-pad ink and analyzing the formation time of documents.

Responses and comprehensive evaluation of growth characteristics of ephemeral plants in the desert-oasis ecotone to soil types.

The ecological environment of the Gurbantünggüt desert-oasis ecotone is extremely fragile. Ephemeral plants are an important part of the ecosystem and play an essential role in maintaining the ecological stability of the ecotone. However, few studies have focused on the growth, soil quality and system sustainability of ephemeral plants in different soils. This study was based on two typical soil types (grey desert soil, GS; aeolian soil, AS) in the aforementioned ecotone, considered four ephemeral plants (Tetracme recurvata, TR; Tetracme contorta, TC; Malcolmia scorpioides, MS; Isatis violascens, IV) as the research object, analysed plant characteristics and soil properties, and comprehensively evaluated the ephemeral plant system by analysing the soil quality index (SQI) and sustainability index (SI). The results showed that there were significant differences in biomass and nutrient accumulation between different ephemeral plants, which were significantly affected by soil types. In the two examined soils (GS and AS), the contents of nutrients and microbial carbon (MBC) and nitrogen (MBN) in the rhizosphere soil were higher than those in the bare soil (BS), and there were significant differences among different species. The key soil factors related to total biomass in GS and AS were also different. The SQI of ephemeral plants was significantly higher than that of the BS, and varied with soil types and plant species. The species with the highest SQI of the key factor data set in GS and AS were IV and TR, respectively. The SI analysis indicated that IV in GS and MS and IV in AS were sustainable, and the plant properties can be better used to assess the sustainability of ephemeral plant systems. In conclusion, ephemeral plants improved the soil quality and system sustainability of the study ecotone. Further, the growth of ephemeral plant and rhizosphere soil properties vary with plant species and soil types; thus, selecting suitable species for large-scale planting in different soil types is of great significance for improving the ecological stability of the ecotone.

Succession of endophytic fungi and arbuscular mycorrhizal fungi associated with the growth of plant and their correlation with secondary metabolites in the roots of plants.

BACKGROUND: To decipher the root and microbial interaction, secondary metabolite accumulation in roots and the microbial community's succession model during the plant's growth period demands an in-depth investigation. However, till now, no comprehensive study is available on the succession of endophytic fungi and arbuscular mycorrhizal fungi (AMF) with roots of medicinal licorice plants and the effects of endophytic fungi and AMF on the secondary metabolite accumulation in licorice plant's root. RESULTS: In the current study, interaction between root and microbes in 1-3 years old medicinal licorice plant's root and rhizospheric soil was investigated. Secondary metabolites content in licorice root was determined using high-performance liquid chromatography (HPLC). The composition and diversity of endophytic and AMF in the root and soil were deciphered using high-throughput sequencing technology. During the plant's growth period, as compared to AMF, time and species significantly affected the diversity and richness of endophytic fungi, such as Ascomycota, Basidiomycota, Fusarium, Cladosporium, Sarocladium. The growth period also influenced the AMF diversity, evident by the significant increase in the relative abundance of Glomus and the significant decrease in the relative abundance of Diversispora. It indicated a different succession pattern between the endophytic fungal and AMF communities. Meanwhile, distance-based redundancy analysis and Mantel tests revealed root's water content and secondary metabolites (glycyrrhizic acid, liquiritin, and total flavonoids), which conferred endophytic fungi and AMF diversity. Additionally, plant growth significantly altered soil's physicochemical properties, which influenced the distribution of endophytic fungal and AMF communities. CONCLUSIONS: This study indicated a different succession pattern between the endophytic fungal and AMF communities. During the plant's growth period, the contents of three secondary metabolites in roots increased per year, which contributed to the overall differences in composition and distribution of endophytic fungal and AMF communities. The endophytic fungal communities were more sensitive to secondary metabolites than AMF communities. The current study provides novel insights into the interaction between rhizospheric microbes and root exudates.

High-throughput sequencing analysis of the rhizosphere arbuscular mycorrhizal fungi (AMF) community composition associated with Ferula sinkiangensis.

BACKGROUND: Ferula sinkiangensis is an increasingly endangered medicinal plant. Arbuscular mycorrhiza fungi (AMF) are symbiotic microorganisms that live in the soil wherein they enhance nutrient uptake, stress resistance, and pathogen defense in host plants. While such AMF have the potential to contribute to the cultivation of Ferula sinkiangensis, the composition of AMF communities associated with Ferula sinkiangensis and the relationship between these fungi and other pertinent abiotic factors still remains to be clarified. RESULTS: Herein, we collected rhizosphere and surrounding soil samples at a range of depths (0-20, 20-40, and 40-60 cm) and a range of slope positions (bottom, middle, top). These samples were then subjected to analyses of soil physicochemical properties and high-throughput sequencing (Illumina MiSeq). We determined that Glomus and Diversispora species were highly enriched in all samples. We further found that AMF diversity and richness varied significantly as a function of slope position, with this variation primarily being tied to differences in relative Glomus and Diversispora abundance. In contrast, no significant relationship was observed between soil depth and overall AMF composition, although some AMF species were found to be sensitive to soil depth. Many factors significantly affected AMF community composition, including organic matter content, total nitrogen, total potassium, ammonium nitrogen, nitrate nitrogen, available potassium, total dissolvable salt levels, pH, soil water content, and slope position. We further determined that Shannon diversity index values in these communities were positively correlated with total phosphorus, nitrate-nitrogen levels, and pH values (P < 0.05), whereas total phosphorus, total dissolvable salt levels, and pH were positively correlated with Chao1 values (P < 0.05). CONCLUSION: In summary, our data revealed that Glomus and Diversispora are key AMF genera found within Ferula sinkiangensis rhizosphere soil. These fungi are closely associated with specific environmental and soil physicochemical properties, and these soil sample properties also differed significantly as a function of slope position (P < 0.05). Together, our results provide new insights regarding the relationship between AMF species and Ferula sinkiangensis, offering a theoretical basis for further studies of their development.

Root-associated endophytic bacterial community composition and structure of three medicinal licorices and their changes with the growing year.

BACKGROUND: The dried roots and rhizomes of medicinal licorices are widely used worldwide as a traditional medicinal herb, which are mainly attributed to a variety of bioactive compounds that can be extracted from licorice root. Endophytes and plants form a symbiotic relationship, which is an important source of host secondary metabolites. RESULTS: In this study, we used high-throughput sequencing technology and high-performance liquid chromatography to explore the composition and structure of the endophytic bacterial community and the content of bioactive compounds (glycyrrhizic acid, liquiritin and total flavonoids) in different species of medicinal licorices (Glycyrrhiza uralensis, Glycyrrhiza glabra, and Glycyrrhiza inflata) and in different planting years (1-3 years). Our results showed that the contents of the bioactive compounds in the roots of medicinal licorices were not affected by the species, but were significantly affected by the main effect growing year (1-3) (P < 0.05), and with a trend of stable increase in the contents observed with each growing year. In 27 samples, a total of 1,979,531 effective sequences were obtained after quality control, and 2432 effective operational taxonomic units (OTUs) were obtained at 97% identity. The phylum Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes, and the genera unified-Rhizobiaceae, Pseudomonas, Novosphingobium, and Pantoea were significantly dominant in the 27 samples. Distance-based redundancy analysis (db-RDA) showed that the content of total flavonoids explained the differences in composition and distribution of endophytic bacterial communities in roots of cultivated medicinal liquorices to the greatest extent. Total soil salt was the most important factor that significantly affected the endophytic bacterial community in soil factors, followed by ammonium nitrogen and nitrate nitrogen. Among the leaf nutrition factors, leaf water content had the most significant effect on the endophytic bacterial community, followed by total phosphorus and total potassium. CONCLUSIONS: This study not only provides information on the composition and distribution of endophytic bacteria in the roots of medicinal licorices, but also reveals the influence of abiotic factors on the community of endophytic bacteria and bioactive compounds, which provides a reference for improving the quality of licorice.

High-throughput sequencing reveals rhizosphere fungal community composition and diversity at different growth stages of Populus euphratica in the lower reaches of the Tarim River.

Background: Populus euphratica is one of the most ancient and primitive tree species of Populus spp and plays an important role in maintaining the ecological balance in desert areas. To decipher the diversity, community structure, and relationship between rhizosphere fungi and environmental factors at different growth stages of P. euphratica demands an in-depth investigation. Methods: In this study, P. euphratica at different growth stages (young, medium, overripe, and decline periods) was selected as the research object, based on the determination of the physicochemical properties of its rhizosphere soil, the fungal community structure and diversity of P. euphratica and their correlation with soil physicochemical properties were comprehensively analyzed through high-throughput sequencing technology (internal transcribed spacer (ITS)) and bioinformatics analysis methods. Results: According to the analysis of OTU annotation results, the rhizosphere soil fungal communities identified in Populus euphratica were categorized into10 phyla, 36 classes, 77 orders, 165 families, 275 genera and 353 species. The alpha diversity analysis showed that there was no obvious change between the different growth stages, while beta diversity analysis showed that there were significantly differences in the composition of rhizosphere soil fungal communities between mature and overripe trees (R 2 = 0.31, P = 0.001), mature and deadwood (R 2 = 0.28, P = 0.001). Ascomycota and Basidiomycota were dominant phyla in the rhizosphere fungal community and the dominant genera were Geopora, Chondrostereum and unidentified_Sordariales_sp. The relative abundance of the top ten fungi at each classification level differed greatly in different stages. Canonical correspondence analysis (CCA) and Spearman's correlation analysis showed that conductivity (EC) was the main soil factor affecting the composition of Populus euphratica rhizosphere soil fungal community (P < 0.01), followed by total dissolvable salts (TDS) and available potassium (AK) (P < 0.05). Conclusions: Our data revealed that the rhizosphere fungal communities at the different growth stages of P. euphratica have differences, conductivity (EC) was the key factor driving rhizosphere fungi diversity and community structure, followed by total dissolvable salts (TDS) and available potassium (AK).

Soil properties, root morphology and physiological responses to cotton stalk biochar addition in two continuous cropping cotton field soils from Xinjiang, China.

Long-term and widespread cotton production in Xinjiang, China, has resulted in significant soil degradation, thereby leading to continuous cropping obstacles; cotton stalk biochar (CSB) addition may be an effective countermeasure to this issue, with effects that are felt immediately by root systems in direct contact with the soil. In this study, we assess the effects of different CSB application rates on soil nutrient contents, root morphology, and root physiology in two soil types commonly used for cotton production in the region. Compared with CK (no CSB addition), a 1% CSB addition increased total nitrogen (TN), available phosphorus (AP), and organic matter (OM) by 13.3%, 7.2%, and 50% in grey desert soil, respectively , and 36.5%, 19.9%, and 176.4%, respectively, in aeolian sandy soil. A 3% CSB addition increased TN, AP, and OM by 38.8%, 23.8%, and 208.1%, respectively, in grey desert soil, and 36%, 13%, and 183.2%, respectively, in aeolian sandy soil. Compared with the aeolian sandy soil, a 1% CSB addition increased TN, OM, and AP by 95%, 94.8%, and 33.3%, respectively, in the grey desert soil , while in the same soil 3% CSB addition increased TN, OM, and AP by 108%, 21.1%, and 73.9%, respectively. In the grey desert soil, compared with CK, a 1% CSB application increased the root length (RL) (34%), specific root length (SRL) (27.9%), and root volume (RV) (32.6%) during the bud stage, increased glutamine synthetase (GS) (13.9%) and nitrate reductase (NR) activities (237%), decreased the RV (34%) and average root diameter (ARD) (36.2%) during the harvesting stage. A 3% CSB addition increased the RL (44%), SRL (20%), and RV (41.2%) during the bud stage and decreased the RV (29%) and ARD (27%) during the harvesting stage. In the aeolian sandy soil, 1% CSB increased the RL (38.3%), SRL (73.7%), and RV (17%), while a 3% caused a greater increase in the RL (55%), SRL (89%), RV (28%), soluble sugar content (128%), and underground biomass (33.8%). Compared with the grey desert soil, a 1% CSB addition increased the RL (48.6%), SRL (58%), and RV (18.6%) in the aeolian sandy soil, while a 3% further increased the RL (54.8%), SRL (84.2%), RV (21.9%), and soluble sugar content (233%). The mechanisms by which CSB addition improves the two soils differ: root morphology changed from coarse and short to fine and long in the grey desert soil, and from fine and long to longer in the aeolian sandy soil. Overall, a 3% CSB addition may be a promising and sustainable strategy for maintaining cotton productivity in aeolian sandy soil in the Xinjiang region.

Differences in the endophytic fungal community and effective ingredients in root of three Glycyrrhiza species in Xinjiang, China.

BACKGROUND: Endophytic fungi influence the quality and quantity of the medicinal plant's bioactive compounds through specific fungus-host interactions. Nevertheless, due to the paucity of information, the composition of endophytic fungal communities and the mechanism by which effective ingredients regulate endophytic fungal communities in roots remains unclear. METHODS: In this study, we collected root and soil samples (depth range: 0-20, 20-40, and 40-60 cm) of three Glycyrrhiza species (Glycyrrhiza uralensis, Glycyrrhiza inflata, and Glycyrrhiza glabra). Glycyrrhizic acid and liquiritin content were determined using high-performance liquid chromatography (HPLC), and total flavonoid content was determined using ultraviolet spectrophotometry. High-throughput sequencing technology was employed to explore the composition and diversity of the endophytic fungal community in different root segments of three Glycyrrhiza species. Furthermore, soil samples were subjected to physicochemical analyses. RESULTS: We observed that the liquiritin content was not affected by the root depth (0-20 cm, 20-40 cm, and 40-60 cm). Still, it was significantly affected by the Glycyrrhiza species (Glycyrrhiza uralensis, Glycyrrhiza inflata, Glycyrrhiza glabra) (P < 0.05). In Glycyrrhiza root, a total of eight phyla and 140 genera were annotated so far, out of which Ascomycota and Basidiomycota phyla, and the Fusarium, Paraphoma, and Helminthosporium genera were found to be significantly dominant. Spearman correlation analysis revealed that liquiritin content was accountable for the differences in the diversity of the endophytic fungal community. Furthermore, distance-based redundancy analysis (db-RDA) showed that physicochemical properties of the soil (available potassium and ammonium nitrogen) and the root factors (liquiritin and water content) were the main contributing factors for the variations in the overall structure of the endophytic fungal community. Our results showed that the effective ingredients of Glycyrrhiza root and physicochemical properties of the soil regulated the endophytic fungal community composition and medicinal licorice diversity.

High throughput sequencing-based analysis of the soil bacterial community structure and functions of Tamarix shrubs in the lower reaches of the Tarim River.

Tamarix is a dominant species in the Tarim River Basin, the longest inland river in China. Tamarix plays an important role in the ecological restoration of this region. In this study, to investigate the soil bacterial community diversity in Tamarix shrubs, we collected soil samples from the inside and edge of the canopy and the edge of nebkhas and non-nebkhas Tamarix shrubs located near the Yingsu section in the lower reaches of Tarim River. High throughput sequencing technology was employed to discern the composition and function of soil bacterial communities in nebkhas and non-nebkhas Tamarix shrubs. Besides, the physicochemical properties of soil and the spatial distribution characteristics of soil bacteria and their correlation were analyzed. The outcomes of this analysis demonstrated that different parts of Tamarix shrubs had significantly different effects on soil pH, total K (TK), available K (AK), ammonium N (NH4 +), and available P (AP) values (P < 0.05), but not on soil moisture (SWC), total salt (TDS), electrical conductivity (EC), organic matter (OM), total N (TN), total P (TP), and nitrate N (NO3 -) values. The soil bacterial communities identified in Tamarix shrubs were categorized into two kingdoms, 71 phyla, 161 classes, 345 orders, 473 families, and 702 genera. Halobacterota, unidentified bacteria, and Proteobacteria were found to be dominant phyla. The correlation between the soil physicochemical factors and soil bacterial community was analyzed, and as per the outcomes OM, AK, AP, EC, and NH4 + were found to primarily affect the structure of the soil bacterial community. SWC, TK and pH were positively correlated with each other, but negatively correlated with other soil factors. At the phyla level, a significantly positive correlation was observed between the Halobacterota and AP, OM as well as Bacteroidota and AK (P < 0.01), but a significantly negative correlation was observed between the Chloroflexi and AK, EC (P < 0.01). The PICRUSt software was employed to predict the functional genes. A total of 6,195 KEGG ortholog genes were obtained. The function of soil bacteria was annotated, and six metabolic pathways in level 1, 41 metabolic pathways in level 2, and 307 metabolic pathways in level 3 were enriched, among which the functional gene related to metabolism, genetic information processing, and environmental information processing was found to have the dominant advantage. The results showed that the nebkhas and canopy of Tamarix shrubs had a certain enrichment effect on soil nutrients content, and bacterial abundance and significant effects on the structure and function of the soil bacterial community.

Variation of rhizosphere bacterial community diversity in the desert ephemeral plant Ferula sinkiangensis across environmental gradients.

Ferula sinkiangensis (F. sinkiangensis) is a desert short-lived medicinal plant, and its number is rapidly decreasing. Rhizosphere microbial community plays an important role in plant growth and adaptability. However, F. sinkiangensis rhizosphere bacterial communities and the soil physicochemical factors that drive the bacterial community distribution are currently unclear. On this study, based on high-throughput sequencing, we explored the diversity, structure and composition of F. sinkiangensis rhizosphere bacterial communities at different slope positions and soil depths and their correlation with soil physicochemical properties. Our results revealed the heterogeneity and changed trend of F. sinkiangensis rhizosphere bacterial community diversity and abundance on slope position and soil depth and found Actinobacteria (25.5%), Acidobacteria (16.9%), Proteobacteria (16.6%), Gemmatimonadetes (11.5%) and Bacteroidetes (5.8%) were the dominant bacterial phyla in F. sinkiangensis rhizosphere soil. Among all soil physicochemical variables shown in this study, there was a strong positive correlation between phosphorus (AP) and the diversity of rhizosphere bacterial community in F. sinkiangensis. In addition, Soil physicochemical factors jointly explained 24.28% of variation in F. sinkiangensis rhizosphere bacterial community structure. Among them, pH largely explained the variation of F. sinkiangensis rhizosphere bacterial community structure (5.58%), followed by total salt (TS, 5.21%) and phosphorus (TP, 4.90%).

High-throughput sequencing reveals the diversity and community structure of rhizosphere fungi of Ferula Sinkiangensis at different soil depths.

Ferula sinkiangesis is a valuable medicinal plant that has become endangered. Improving the soil habitat of Ferula sinkiangesis can alleviate plant damage. Fungi play an important role in the soil, but current information on the fungal community structure in the habitat of Ferula sinkiangesis and the relationship between soil fungi and abiotic factors remains unclear. In this study, we analyzed the relative abundance of fungal species in the rhizosphere of Ferula sinkiangesis. Spearman correlation analysis showed that the abiotic factor total potassium (TK) significantly explained the alpha diversity of the fungal community. At altitude, available phosphorus (AP), nitrate nitrogen (NN) and TK were significantly associated with the fungal species. In addition, a two-way ANOVA showed that soil depth had no significant effects on the alpha diversity of rhizosphere and non-rhizosphere fungi. Interestingly, linear discriminant effect size (LEfSe) analysis indicated that different biomarkers were present at varying soil depths. These findings may be related to the growth and medicinal properties of Ferula Sinkiangensis.

Future N deposition and precipitation changes will be beneficial for the growth of Haloxylon ammodendron in Gurbantunggut Desert, northwest China.

Evaluation of precipitation and nitrogen (N) deposition in desert ecosystems helps to elucidate the reaction of desert ecosystems to future environmental changes. An in-situ field experiment was established to examine the influence of a long-term enhanced precipitation and N deposition on the photosynthetic traits and physiological characteristics of Haloxylon ammodendron in the Gurbantunggut Desert, northwest China, throughout the growing season in 2014-2016. Results showed a significant interaction between precipitation and N applications. Increased precipitation and N deposition and their coupling could significantly improve photosynthetic capacity, alter the variability in amplitude of water potential and change the content of substances regulating osmotic pressure in H. ammodendron. According to the comprehensive evaluation of H. ammodendron's adaptability using six different water and N coupling models, a combination of a 30% increase in precipitation and a 30 kg N ha-1 yr-1 addition in nitrogen deposition, or the addition of N at a concentration of 60 kg N ha-1 yr-1 with natural precipitation were beneficial to H. ammodendron growth and development. Hence, changes in the future global environment can be anticipated to be beneficial to H. ammodendron growth.

Bacterial diversity and community structure in the rhizosphere of four Ferula species.

The medicinal value of the Ferula L. has been recognized for more than a thousand years. Wild stocks of Ferula have declined dramatically because high economic value has led to overharvesting. The objective of this study was to compare the rhizosphere microbial community of four Ferula species [F. syreitschikowii K.-Pol., F. gracilis (Ledeb.) Ledeb., F. ferulaeoides (Steud.) Korov., and F. lehmannii Boiss.] in the northern part of Xinjiang, China. The 16S rRNA sequences of rhizosphere bacteria were obtained with an Illumina paired-end sequence platform. Analysis was conducted to determine the richness and diversity of the rhizosphere bacterial communities. Two-way ANOVA indicated that plant species and soil depth had no significant effect on the alpha diversity of rhizobacteria. Linear discriminant analysis effect size showed that F. lehmannii followed by F. ferulaeoides had the most biomarkers and the highest taxon level, F. syreitschikowii and F. gracilis the least, while F. syreitschikowii and F. gracilis had the least property. This trend is consistent with reports that the medicinal value of F. lehmannii and F. ferulaeoides is greater than that of F. gracilis and F. syreitschikowii. The results of this study provide information that could be used for the commercial cultivation of Ferula spp.

The effects of neuromuscular blockade on operating conditions during general anesthesia for spinal surgery.

BACKGROUND: Muscle relaxants are prescribed routinely for patients undergoing general anesthesia, but the requirement for paralysis in spinal surgery is unclear. This study compared the operating conditions of general anesthesia with and without a muscle relaxant on spinal surgery patients. METHODS: Eighty-six adults who underwent elective spinal surgery were randomly assigned to a relaxant group (group R) or a no-relaxant group (group NR). All patients were induced with intravenous midazolam (0.05 mg/kg), fentanyl (4 µg/kg), propofol (1.0 mg/kg), and succinylcholine (2 mg/kg) and then atracurium was used in group R but not in group NR. The operating conditions, including muscle tone, body movements, airway pressure, anesthetics consumption, eye-opening time, extubation time, and the Observer's Assessment of the Alertness/Sedation (OAA/S) score 20 minutes after the extubation were compared between the 2 groups. RESULTS: The operating conditions including muscle tone scales, body movements, and airway pressure did not differ between the 2 groups. Eye-opening time (9.35±2.34 vs. 11.02±2.50 min; P=0.002) and extubation time (13.95±3.41 vs. 16.72±3.67 min; P=0.001) were shorter in group NR than in group R. The BIS score at extubation (87.2±5.0 vs. 83.3±5.7; P=0.001) and the OAA/S score 20 minutes after extubation (5 [3 to 5] vs. 4 [3 to 5]; P=0.005) were significantly higher in group NR than in group R. Propofol consumption was higher in group NR than in group R (4206.10±415.80 vs. 3900.60±365.40 µg/kg, respectively; P=0.001). CONCLUSIONS: General anesthesia without muscle relaxant provides similar working conditions to those observed with muscle relaxant, and it is associated with earlier eye opening and extubation and higher level of consciousness on emergence from spinal surgery.

Preparative separation and purification of sulforaphene from radish seeds by high-speed countercurrent chromatography.

Sulforaphene, a kind of isothiocyanates, derived from glucoraphenin which is the important ingredient of radish (Raphanus sativus L.) seeds, has shown significant pharmacological activities. In this paper, the separation and purification of sulforaphene from radish seeds, was achieved by high-speed countercurrent chromatography (HSCCC). A two-phase solvent system consisted of n-hexane-ethyl acetate-methanol-water (35:100:35:100, v/v/v/v) was applied. The revolution speed of the separation column, flow rate of the mobile phase and separation temperature were 800 rpm, 2 ml/min and 30°C, respectively. From about 1000 mg amount of the crude plant extract, 249.4 mg of pure sulforaphene was obtained by one-step separation on a 280 ml HSCCC column. The purified sulforaphene was at a high purity of 96.9% and the mass recovery was more than 95%. The purity of sulforaphene was determined by HPLC analysis and its chemical structure was assessed by MS, (1)H NMR, (13)C NMR and DEPT-135 NMR.

Separation and purification of sulforaphene from radish seeds using macroporous resin and preparative high-performance liquid chromatography.

This present study described a rapid and cost-effective method for the separation and purification of natural sulforaphene from radish seeds by SP-700 macroporous resin and preparative high-performance liquid chromatography (HPLC). Sulforaphene with high purity and recovery was obtained by preparative HPLC with a C18 column and 30% methanol in ultra-pure water as the mobile phase. 12.5 kg of radish seeds, which contained 87.5 g of sulforaphene, produced 117.5 g of sulforaphene-rich extract of 65.8% sulforaphene after primary separation by SP-700 macroporous resin. 5.9 g of 96.5% sulforaphene was obtained from 9.5 g of the sulforaphene-rich extract after purification by preparative HPLC. The purified compound was assessed by analytical HPLC and characterised by ESI/MS, (1)H NMR and (13)C NMR. Standard curve was developed using the purified sulforaphene to allow quantification of sulforaphene in the extracts of radish seeds by analytical HPLC.

Development and validation of a new PCR optimization method by combining experimental design and artificial neural network.

Polymerase chain reaction (PCR) is one of the most powerful techniques in a variety of clinical and biological research fields. In this paper, a chemometrics approach, combining experimental design (ED) and artificial neural network (ANN), was proposed for optimization of PCR amplification of lycopene cyclase gene carRA in Blakeslea Trispora. Five-level star design was carried out to obtain experimental information and provide data source for ANN modeling. Nine variables were used as inputs in ANN, including the added amount of template, primer, dNTP, polymerase and magnesium ion, the temperature of denaturating, annealing and extension, and the number of cycles. The output variable was the efficiency (yield) of the PCR. Based on the developed model, the effects of each parameter on PCR efficiency were predicted and the most suitable operation condition for present system was determined. At last, the validation experiment was performed under the optimized condition, and the expectant results were produced. The results obtained in this paper showed that the combination of ANN and ED provided a satisfactory optimization model with good descriptive and predictive abilities, indicating that the method of combining ANN and ED can be a useful tool in PCR optimization and other biological applications.