Genome-wide identification and expression analysis of the KNOX family and its diverse roles in response to growth and abiotic tolerance in sweet potato and its two diploid relatives.

KNOXs, a type of homeobox genes that encode atypical homeobox proteins, play an essential role in the regulation of growth and development, hormonal response, and abiotic stress in plants. However, the KNOX gene family has not been explored in sweet potato. In this study, through sequence alignment, genomic structure analysis, and phylogenetic characterization, 17, 12 and 11 KNOXs in sweet potato (I. batatas, 2n = 6x = 90) and its two diploid relatives I. trifida (2n = 2x = 30) and I. triloba (2n = 2x = 30) were identified. The protein physicochemical properties, chromosome localization, phylogenetic relationships, gene structure, protein interaction network, cis-elements of promoters, tissue-specific expression and expression patterns under hormone treatment and abiotic stresses of these 40 KNOX genes were systematically studied. IbKNOX4, -5, and - 6 were highly expressed in the leaves of the high-yield varieties Longshu9 and Xushu18. IbKNOX3 and IbKNOX8 in Class I were upregulated in initial storage roots compared to fibrous roots. IbKNOXs in Class M were specifically expressed in the stem tip and hardly expressed in other tissues. Moreover, IbKNOX2 and - 6, and their homologous genes were induced by PEG/mannitol and NaCl treatments. The results showed that KNOXs were involved in regulating growth and development, hormone crosstalk and abiotic stress responses between sweet potato and its two diploid relatives. This study provides a comparison of these KNOX genes in sweet potato and its two diploid relatives and a theoretical basis for functional studies.

Comparative analysis of main bio-active components in the herb pair Danshen-Honghua and its single herbs by ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry.

A sensitive, reliable, and powerful ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry method was developed for simultaneous quantification of the 15 main bio-active components including phenolic acids and flavonoids within 13 min for the first time. The proposed method was first reported and validated by good linearity (r2  > 0.9975), limit of detection (1.12-7.01 ng/mL), limit of quantification (3.73-23.37 ng/mL), intra- and inter-day precisions (RSD ≤ 1.92%, RSD ≤ 2.45%), stability (RSD ≤ 5.63%), repeatability (RSD ≤ 4.34%), recovery (96.84-102.12%), and matrix effects (0.92-1.02). The established analytical methodology was successfully applied to comparative analysis of main bio-active components in the herb pair Danshen-Honghua and its single herbs. Compared to the single herb, the content of most flavonoid glycosides was remarkably increased in their herb pair, and main phenolic acids were decreased, conversely. The content changes of the main components in the herb pair supported the synergistic effects on promoting blood circulation and removing blood stasis. The results provide a scientific basis and reference for the quality control of Danshen-Honghua herb pair and the drug interactions based on variation of bio-active components in herb pairs.

[Comparative study on promoting blood effects of Danshen-Honghua herb pair with different preparations based on chemometrics and multi-attribute comprehensive index methods].

To evaluate the promoting blood circulation and removing blood stasis effects of Danshen-Honghua(DH) herb pair with different preparations (alcohol, 50% alcohol and water) on blood rheology and coagulation functions in acute blood stasis rats, and optimize the best preparation method of DH based on principal component analysis(PCA), hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods. Ice water bath and subcutaneous injection of adrenaline were both used to establish the acute blood stasis rat model. Then the blood stasis rats were administrated intragastrically with DH (alcohol, 50% alcohol and water) extracts. The whole blood viscosity(WBV), plasma viscosity(PV), erythrocyte sedimentation rate(ESR) and haematocrit(HCT) were tested to observe the effects of DH herb pair with different preparations and doses on hemorheology of blood stasis rats; the activated partial thromboplastin time(APTT), thrombin time(TT), prothrombin time(PT), and plasma fibrinogen(FIB) were tested to observe the effects of DH herb pair with different preparations on blood coagulation function and platelet aggregation of blood stasis rats. Then PCA, hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods were all used to comprehensively evaluate the total promoting blood circulation and removing blood stasis effects of DH herb pair with different preparations. The hemorheological indexes and coagulation parameters of model group had significant differences with normal blank group. As compared with the model group, the DH herb pair with different preparations at low, middle and high doses could improve the blood hemorheology indexes and coagulation parameters in acute blood stasis rats with dose-effect relation. Based on the PCA, hierarchical cluster heatmap analysis and multi-attribute comprehensive index methods, the high dose group of 50% alcohol extract had the best effect of promoting blood circulation and removing blood stasis. Under the same dose but different preparations, 50% alcohol DH could obviously improve the hemorheology and blood coagulation function in acute blood stasis rats. These results suggested that DH herb pair with different preparations could obviously ameliorate the abnormality of hemorheology and blood coagulation function in acute blood stasis rats, and the optimized preparation of DH herb pair on promoting blood effects was 50% alcohol extract, providing scientific basis for more effective application of the DH herb pair in modern clinic medicine.

Comparative Analysis of the Effects of Hydroxysafflor Yellow A and Anhydrosafflor Yellow B in Safflower Series of Herb Pairs Using Prep-HPLC and a Selective Knock-Out Approach.

The flower of Carthamus tinctorius L. (Carthami Flos, safflower), important in traditional Chinese medicine (TCM), is known for treating blood stasis, coronary heart disease, hypertension, and cerebrovascular disease in clinical and experimental studies. It is widely accepted that hydroxysafflor yellow A (HSYA) and anhydrosafflor yellow B (ASYB) are the major bioactive components of many formulae comprised of safflower. In this study, selective knock-out of target components such as HSYA and ASYB by using preparative high performance liquid chromatography (prep-HPLC) followed by antiplatelet and anticoagulation activities evaluation was used to investigate the roles of bioactive ingredients in safflower series of herb pairs. The results showed that both HSYA and ASYB not only played a direct role in activating blood circulation, but also indirectly made a contribution to the total bioactivity of safflower series of herb pairs. The degree of contribution of HSYA in the safflower and its series herb pairs was as follows: Carthami Flos-Ginseng Radix et Rhizoma Rubra (CF-GR) > Carthami Flos-Sappan Lignum (CF-SL) > Carthami Flos-Angelicae Sinensis Radix (CF-AS) > Carthami Flos-Astragali Radix (CF-AR) > Carthami Flos-Angelicae Sinensis Radix (CF-AS) > Carthami Flos-Glycyrrhizae Radix et Rhizoma (CF-GL) > Carthami Flos-Salviae Miltiorrhizae Radix et Rhizoma (CF-SM) > Carthami Flos (CF), and the contribution degree of ASYB in the safflower and its series herb pairs: CF-GL > CF-PS > CF-AS > CF-SL > CF-SM > CF-AR > CF-GR > CF. So, this study provided a significant and effective approach to elucidate the contribution of different herbal components to the bioactivity of the herb pair, and clarification of the variation of herb-pair compatibilities. In addition, this study provides guidance for investigating the relationship between herbal compounds and the bioactivities of herb pairs. It also provides a scientific basis for reasonable clinical applications and new drug development on the basis of the safflower series of herb pairs.

[Rapidly Detection for Moso Bamboo Density under Different Moisture ConditionBased on X-CT Technology].

Density, which is closely relate with many physical and mechanical properties of bamboo, is one of the important indicators of bamboo material properties. Moreover, because of existing different moisture gradients in bamboo, the measured results of the density are different. Based on X-ray computed tomography (X-CT) technology, the divergent degree of the CT values of 7 different aged Moso bamboo was compared under oven-dried, air-dried and water-saturated conditions. Except for the 4-year-old and 10-year-old Moso bamboo, the CT values of other aged bamboos have minor differences with each other; the models for the measured CT values and the corresponding densities of Moso bamboo were respectively fitted under oven-dried, air-dried and water-saturated conditions. Meanwhile, the model was also fitted under different moisture gradients, which was composed by the measured CT values and the corresponding densities of Moso bamboo. Then the relations between the CT values andthe densitiesof 7 different aged Moso bamboo were systematically analyzed under single moisture content and three moisture gradients;the CT values were fitted under oven-dried condition, of which the radial positions are relative to the outer of Moso bamboo. According to the relation between the CT value and the density, the fitting curves explain the reasons for the radial density variations of 7 different aged Moso bamboo. Results show that the relations, which are fitted by the measured densities and the corresponding CT values of 7 different aged Moso bamboo under oven-dried, air-dried and water-saturated conditions, are good linear and the slopes of those models are approximate; the relation of the densities with the CT values for Moso bamboo is linear under different moisture gradients, moreover, which is rarely affected by moisture. The regression equation is: D=0.001 H+1.003 2, R2=0.968 3(D is the density, H is the CT value) and the determination coefficient of the validation model is: R2=0.974 3; there is no obvious variation between the densities of the inner and the outer, but not in middle part to 7 different aged Moso bamboo under oven-dried condition. To realize rapid detection on the densities of Moso bamboo under different moisture content, these results provide technical support and data reference. At the same time, X-ray computed tomography (X-CT) technology also puts forward a new feasible way for the further studies of bamboo material properties and structure.

[Variation of microfibril angle in Dendrocalamus farinosus analyzed based on X-ray diffraction spectrum and its effect on tensile properties].

X-ray diffraction technology was used to rapidly predict variation in microfibril angle (MFA) in Dendrocalamus fari- X-ray diffraction technology was used to rapidly predict variation in microfibril angle (MFA) in Dendrocalamus farinosus. The results show that there is little variation in MFA with bamboo age from 2a to 5e, and MFA of 3a is at the maximum with the value of 8.521 degrees. The average value of MFA of 2a or 3a is greater than 4a or 5a with absolute differences less than 0.10 degrees. MFA of base, middle and upper position respectively are 8.499 degrees, 8.497 degrees and 8.483 degrees with coefficient of variation about 5%. There is an increasing tendency from the periphery to the inner of bamboo culms. Variance analysis indicates that MFA is highly sensitive to radial position, but insensitive to bamboo age and longitudinal position. The correlation coefficient of longitudinal strength and modulus of elasticity (MOE) is 0.57. MFA was responsible for 35% and 43% of the variation found in longitudinal strength and MOE respectively, which means MFA has a certain extent effect on mechanical properties.

Specification and guideline for technical aspects and scanning parameter settings of neonatal lung ultrasound examination.

Lung ultrasound (LUS) is now widely used in the diagnosis and monitor of neonatal lung diseases. Nevertheless, in the published literatures, the LUS images may display a significant variation in technical execution, while scanning parameters may influence diagnostic accuracy. The inter- and intra-observer reliabilities of ultrasound exam have been extensively studied in general and in LUS. As expected, the reliability declines in the hands of novices when they perform the point-of-care ultrasound (POC US). Consequently, having appropriate guidelines regarding to technical aspects of neonatal LUS exam is very important especially because diagnosis is mainly based on interpretation of artifacts produced by the pleural line and the lungs. The present work aimed to create an instrument operation specification and parameter setting guidelines for neonatal LUS. Technical aspects and scanning parameter settings that allow for standardization in obtaining LUS images include (1) select a high-end equipment with high-frequency linear array transducer (12-14 MHz). (2) Choose preset suitable for lung examination or small organs. (3) Keep the probe perpendicular to the ribs or parallel to the intercostal space. (4) Set the scanning depth at 4-5 cm. (5) Set 1-2 focal zones and adjust them close to the pleural line. (6) Use fundamental frequency with speckle reduction 2-3 or similar techniques. (7) Turn off spatial compounding imaging. (8) Adjust the time-gain compensation to get uniform image from the near-to far-field.

Herb-drug interactions: in vivo and in vitro effect of Shenmai injection, a herbal preparation, on the metabolic activities of hepatic cytochrome P450 3A1/2, 2C6, 1A2, and 2E1 in rats.

Shenmai injection (SMI), a mixture of Radix Ginseng and Radix Ophiopogonis, is one of the most popular herbal medicinal products and is widely used for the treatment of coronary atherosclerotic cardiopathy and viral myocarditis. The purpose of this study was to investigate the effect of SMI, in vivo and in vitro, on the metabolic activities of hepatic cytochrome CYP450 3A1/2, 2C6, 2E1, and 1A2 in rats. After a single or multiple pretreatment with SMI, the rats were administrated intravenously a cocktail containing midazolam (1 mg/kg), diclofenac (0.5 mg/kg), theophylline (1 mg/kg), and chlorzoxazone (0.5 mg/kg) as probe substrates of rat CYP450 3A1/2, 2C6, 1A2, and 2E1, respectively. Single and multiple SMI pretreatment to rats resulted in a rise of 33.8 % (p < 0.01) and 25.6 % (p < 0.01) in AUC for midazolam, and an increase in AUC for diclofenac by 14.7 % (p < 0.05) and 31.2 % (p < 0.01), respectively. However, the pharmacokinetics of chlorzoxazone and theophylline in rats was not altered markedly. In rat liver microsomes, linear mixed-type inhibition of SMI against the enzyme activities of CYP3A1/2, CYP2C6, and CYP1A2 was shown with IC(50) values of 3.3 %, 2.0 %, and 3.1 % and K(i) values of 3.8 %, 1.5 %. and 1.9 %, respectively. These in vivo and in vitro results demonstrated that SMI had the potential to inhibit the activities of hepatic CYP3A1/2 and CYP2C6, but might not significantly affect CYP1A2 and CYP2E1-mediated metabolism in rats.

Cross electro-nape-acupuncture ameliorates cerebral hemorrhage-induced brain damage by inhibiting necroptosis.

BACKGROUND: Receptor interacting protein kinase 1 (RIPK1)-mediated cell death, including apoptosis and necroptosis, belongs to programmed cell death. It has been reported that RIPK1-mediated necroptosis exists in lesions of cerebral hemorrhage (CH). Electroacupuncture, a treatment derived from traditional Chinese medicine, could improve neurological impairment in patients with brain injury. AIM: To investigate the protective role of cross electro-nape acupuncture (CENA) in CH, and clarify the potential mechanism. METHODS: CH rat models were established, and CENA was applied to the experimental rats. Neurological functions and encephaledema were then measured. Necrotic cells in the brain of rats with CH were evaluated by propidium iodide staining. Necroptosis was assessed by immunofluorescence. Activation of the necroptosis-related pathway was detected by western blot. Extraction of brain tissue, cerebrospinal fluid and serum samples was conducted to measure the expression and secretion of inflammatory cytokines by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: The necroptotic marker p-MLKL was detectable in the brains of rats with CH. Next, we found that CENA could ameliorate neurological functions in rat models of CH. Moreover, the upregulation of RIPK1-mediated necroptosis-related molecules in the brains of rats with CH were inhibited by CENA. Further investigation revealed that CENA partially blocked the interaction between RIPK1 and RIPK3. Finally, in vivo assays showed that CENA decreased the expression of the inflammatory cytokines tumor necrosis factor-α, interleukin-6 and interleukin-8 in CH rat models. CONCLUSION: These findings revealed that CENA exerts a protective role in CH models by inhibiting RIPK1-mediated necroptosis.

International Expert Consensus and Recommendations for Neonatal Pneumothorax Ultrasound Diagnosis and Ultrasound-guided Thoracentesis Procedure.

Pneumothorax (PTX) represents accumulation of the air in the pleural space. A large or tension pneumothorax can collapse the lung and cause hemodynamic compromise, a life-threatening disorder. Traditionally, neonatal pneumothorax diagnosis has been based on clinical images, auscultation, transillumination, and chest X-ray findings. This approach may potentially lead to a delay in both diagnosis and treatment. The use of lung US in diagnosis of PTX together with US-guided thoracentesis results in earlier and more precise management. The recommendations presented in this publication are aimed at improving the application of lung US in guiding neonatal PTX diagnosis and management.

Protocol and Guidelines for Point-of-Care Lung Ultrasound in Diagnosing Neonatal Pulmonary Diseases Based on International Expert Consensus.

Ultrasound is a safe bedside imaging tool that obviates the use of ionizing radiation diagnostic procedures. Due to its convenience, the lung ultrasound has received increasing attention from neonatal physicians. Nevertheless, clear reference standards and guideline limits are needed for accurate application of this diagnostic modality. This document aims to summarize expert opinions and to provide precise guidance to help facilitate the use of the lung ultrasound in the diagnosis of neonatal lung diseases.

A Cross-sectional Survey of Disability Attributed to Mental Disorders and Service Use in China.

BACKGROUND: Mental disorders are strongly associated with disabilities. National survey on disability could provide a reliable basis for policymaking in care and rehabilitation of disabled persons. This study aimed to describe the disability prevalence rates attributed to mental disorders, their distribution by sociodemographic factors, and utilizations of service. METHODS: This study is a secondary data analysis of the Second National Sample Survey on Disability in 2006. The disability and severity were assessed using the World Health Organization Disability Assessment Schedule 2.0. Mental disorders were diagnosed according to the International Statistical Classification of Diseases and Related Health Problems 10th Revision Classification of Mental and Behavioral Disorders. Using descriptive and analytic epidemiological methods, prevalence rates of disability attributed to mental disorders and service use were calculated. RESULTS: Data of 2,526,145 respondents were analyzed. The disability prevalence rate attributed to mental disorders in China was 6.3‰, accounting for 9.9% of all disabled people. Regarding disability prevalence attributed to mental disorders, it showed that gender, residential area, marital status, education level, and economic area were related to the prevalence distributions. The proportions of mild disability were highest in the disabled people with onset age of 18-64 years, while the proportion of extremely severe disability was highest in the disabled people with onset age of 65 years and above. Only 58.6% of disabled people attributed to mental disorders used some of the services. CONCLUSIONS: There are statistical differences of disability prevalence attributed to mental disorders by people and region in China. Service use in disabled people with mental disorders is insufficient.

[Variation characteristics of farmland soil pH in the past 30 years of Enshi Autonomous Prefecture, Hubei, China]. / 恩施州耕地土壤pH近30年变化特征.

In order to explore temporal-spatial variability of farmland soil pH at Enshi Antonomous Prefecture, Hubei, China, soil pH during the past three decades was analyzed, using the datasets of the Second National Soil Survey (1980-1983) and the Cultivated Land Quality Evaluation (2010-2013). The natural and human factors inducing the change of soil pH were evaluated to provide theoretical guidance for further soil acidification management. Results showed that acidic soil (i.e., pH＜6.5) and neutral and alkaline soil (i.e., pH 6.5-8.5) were accounted for 98.4% and 1.6% in the farmland during the period of 2010-2013, respectively. The ratio increased 61.4% for the acidic soil but decreased 61.2% for the neutral and alkaline soil as compared with the period of 1980-1983. In addition, there was no alkaline soil (pH>8.5) in the region in 2010-2013. According to the dataset of the Second National Soil Survey (1980-1983), acidic soil was mainly distributed at Laifeng, Lichuan, Xuanen and Xianfeng counties, with the area ratio of 74.4%, 63.5%, 61.3% and 60.7%, respectively. For the period of 2010-2013, the ratio of acidic soil enhanced widely which was above 96% for each county. At Enshi Autonomous Prefecture, farmland soil showed an obvious acidification trend during the past three decades, with spatial variation of higher in the eastern part and lower in the western part of the region. Furthermore, soil pH decline occurred among different land use types in different areas. Overall, farmland soil pH declined 0.90 on average, with 1.14 decrease for upland and 0.87 for paddy soil, respectively. Clearly, upland soil acidification was severe than paddy soil. Factors related to soil acidification in the Enshi Autonomous Prefecture were mainly human factors such as unreasonable fertilizer combination, fertilizer ratio change, and more base cations taking away by high crop yield.

Comparative metabolomics analysis for the compatibility and incompatibility of kansui and licorice with different ratios by UHPLC-QTOF/MS and multivariate data analysis.

Kansui, the root of Euphorbia kansui T.N. Liou ex T.P. Wang (Euphorbiaceae), is a well-known poisonous traditional Chinese medicine (TCM). However, many monographs of TCM indicated that it cannot be co-used with licorice, as kansui-licorice is a typical "eighteen incompatible" medicaments. Our previous studies have indicated that kansui was effective in treating malignant pleural effusion (MPE), and the efficacy could be weakened by the co-use of licorice, even causing serious toxicity at the given ratio. Nevertheless, the actual mechanisms of their dosage-toxicity-efficacy relationship need to be well clarified. The present study aimed to investigate the effect of individual and combined use of kansui and licorice on MPE rats, and explain the underlying mechanisms from a metabolomic perspective. Urine samples were analyzed by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Partial least-squares discriminate analysis (PLS-DA) models were built to evaluate the interaction between kansui and licorice. Seven potential biomarkers contribute to the separation of model group and control group were tentatively identified. And selenoamino acid metabolism and nicotinate and nicotinamide metabolism with the impact-value 0.31 and 0.24, respectively, were filtered out as the most important metabolic pathways. Kansui and kansui-licorice at a ratio of 4:1 can treat MPE rats by adjusting abnormal metabolic pathways to the normal state, while it may have opposite result with kansui-licorice 1:4. The different influences to the two metabolic pathways may partially explain the dosage-toxicity-efficacy relationship of kansui-licorice with different ratios. The results could offer valuable insights into the compatibility property changes for the two herbs.

A new two-dimensional manganese(II) coordination polymer based on thiophene-3,4-dicarboxylic acid.

A novel manganese coordination polymer, poly[(µ5-thiophene-3,4-dicarboxylato)manganese(II)], [Mn(C6H2O4S)]n, was synthesized hydrothermally using 3,4-thiophenedicarboxylate (3,4-tdc(2-)) as the organic linker. The asymmetric unit of the complex contains an Mn(2+) cation and one half of a deprotonated 3,4-tdc(2-) anion, both residing on a twofold axis. Each Mn(2+) centre is six-coordinated by O atoms of bridging/chelating carboxylate groups from five 3,4-tdc(2-) anions, forming a slightly distorted octahedron. The Mn(2+) centres are bridged by 3,4-tdc(2-) anions to give an infinite two-dimensional layer which incorporates one-dimensional Mn-O gridlike chains, and in which the 3,4-tdc(2-) anion adopts a novel hexadentate chelating and µ5-bridging coordination mode. The fully deprotonated 3,4-tdc(2-) anion exhibits unexpected efficiency as a ligand towards the Mn(2+) centres, which it coordinates through all of its carboxylate O atoms to provide the novel coordination mode. The IR spectrum of the complex is also reported.

Quantitative determination of ophiopogonin d by liquid chromatography/electrospray ionization mass spectrometry and its pharmacokinetics in rat.

A sensitive and rapid liquid chromatography-mass spectrometric method for the determination of ophiopogonin D in rat plasma was developed and validated. Chromatographic separation was performed on a C (18) column using a step gradient program with the mobile phase of 0.5 mmol/L ammonium chloride solution and acetonitrile. Ophiopogonin D was quantified using an electrospray negative ionization mass spectrometry in the selected ion monitoring (SIM) mode using digoxin as an internal standard. Good linearity was obtained in the concentration range of 2.5 - 480.0 ng/mL ( R2 = 0.9984). The lower limit of quantification (LLOQ) and lower limit of detection (LLOD) were 2.5 ng/mL and 1.0 ng/mL, respectively. Both the intra- and inter-day precision was less than 8.9 % and the accuracy was within 97.5 - 107.3 %. The pharmacokinetic study of ophiopogonin D in rats was then defined using the method after intravenous dosing (77.0 microg/kg). The plasma concentration-time profile for ophiopogonin D was best fitted to an open two-compartment model with a clearance of 0.024 +/- 0.010 L/min/kg and a terminal half life of 17.29 +/- 1.70 min. A comparison of the pharmacokinetics of ophiopogonin D as a pure compound and as a component of 'SHENMAI' injection revealed a significantly smaller clearance of ophiopogonin D (0.007 +/- 0.002 L/min/kg) for the latter formulation, consistent with an inhibition by one or more other components in the formulation.