CRISPR/Cas9 targeted mutations of OsDSG1 gene enhanced salt tolerance in rice.

Salinization is one of the leading causes of arable land shrinkage and rice yield decline, recently. Therefore, developing and utilizing salt-tolerant rice varieties have been seen as a crucial and urgent strategy to reduce the effects of saline intrusion and protect food security worldwide. In the current study, the CRISPR/Cas9 system was utilized to induce targeted mutations in the coding sequence of the OsDSG1, a gene involved in the ubiquitination pathway and the regulation of biochemical reactions in rice. The CRISPR/Cas9-induced mutations of the OsDSG1 were generated in a local rice cultivar and the mutant inheritance was validated at different generations. The OsDSG1 mutant lines showed an enhancement in salt tolerance compared to wild type plants at both germination and seedling stages indicated by increases in plant height, root length, and total fresh weight as well as the total chlorophyll and relative water contents under the salt stress condition. In addition, lower proline and MDA contents were observed in mutant rice as compared to wild type plants in the presence of salt stress. Importantly, no effect on seed germination and plant growth parameters was recorded in the CRISRP/Cas9-induced mutant rice under the normal condition. This study again indicates the involvement of the OsDSG1 gene in the salt resistant mechanism in rice and provides a potential strategy to enhance the tolerance of local rice varieties to the salt stress.

Ethynyl-B(dan) in [3+2] Cycloaddition and Larock Indole Synthesis: Synthesis of Stable Boron-Containing Heteroaromatic Compounds.

The cycloaddition of nitrile oxides with ethynyl-B(dan) (dan=naphthalene-1,8-diaminato) allowed the facile preparation of diverse isoxazolyl-B(dan) compounds, all of which displayed excellent protodeborylation-resistant properties. The dan-installation on the boron center proves vital to the high stability of the products as well as the perfect regioselectivity arising from hydrogen bond-directed orientation in the cycloaddition. The diminished boron-Lewis acidity of ethynyl-B(dan) also renders it amenable to azide-alkyne cycloaddition, Larock indole synthesis and related heteroannulations. The obtained boron-containing triazole, indoles, benzofuran and indenone exhibit sufficient resistance toward protodeborylation. Despite the commonly accepted transmetalation-inactive property derived from the diminished Lewis acidity, the synthesized heteroaryl-B(dan) compound was still found to be convertible to the oligoarene via sequential Suzuki-Miyaura coupling.

Multi-omics approaches for the understanding of therapeutic mechanism for Huang-Qi-Long-Dan Granule against ischemic stroke.

After long-term clinical application, traditional Chinese medicine (TCM) has accumulated rich experience in the stroke treatment. Huang-Qi-Long-Dan Granule (HQLDG) is a TCM formula that has been used in clinical for the treatment of acute ischemic stroke. However, its mechanism against ischemic stroke is still unknown. This study aimed to identify HQLDG's effect against ischemic stroke and explore its underlying mechanism. 16s rRNA sequencing, metabolomics/tryptophan (Trp)-targeted metabolomics analysis and transcriptomic analysis were used to investigate HQLDG underlying therapeutic mechanism. Our results revealed that HQLDG significantly decreased the infarct volume, improved mouse behavior and brain slices pathological staining. In addition, it could ameliorate intestinal barrier damage and regulate tight junction gene expression. 16s rRNA, metabolomics and transcriptomics analysis revealed that HQLDG treatment significantly improved the composition of gut microbiota and Trp metabolism pathway, and further downregulated Th17/IL-17 signaling pathway. HQLDG treatment could significantly decrease serum inflammatory cytokines, IL-17A and IL-22; down-regulate Trp metabolism receptor gene (Ahr), inflammatory cytokines genes (IL-17a, IL-22), and an important coding gene for maintaining the mature Th17 (rorc) in both brain and intestinal tissues. In the contrary, after gut microbiota removal, this effect of HQLDG was impaired. HQLDG treated mouse fecal microbiota transplantation also had positive effect against tMCAO injury. Moreover, AhR inhibitor could decrease IL-17A immunofluorescence. These results suggested that the gut microbiota regulation might be an important intermediate in HQLDG against tMCAO injury. HQLDG might exert anti-ischemic stroke effects through the gut microbiota-Trp metabolism-Th17/IL-17 signaling, which provides new insights into HQLDG-mediated prevention in ischemic stroke.

Cost-effectiveness of severe acute malnutrition treatment delivered by community health workers in the district of Mayahi, Niger.

BACKGROUND: A non-randomized controlled trial, conducted from June 2018 to March 2019 in two rural communes in the health district of Mayahi in Niger, showed that including community health workers (CHWs) in the treatment of severe acute malnutrition (SAM) resulted in a better recovery rate (77.2% vs. 72.1%) compared with the standard treatment provided solely at the health centers. The present study aims to assess the cost and cost-effectiveness of the CHWs led treatment of uncomplicated SAM in children 6-59 months compared to the standard national protocol. METHODS: To account for all relevant costs, the cost analysis included activity-based costing and bottom-up approaches from a societal perspective and on a within-trial time horizon. The cost-effectiveness analysis was conducted through a decision analysis network built with OpenMarkov and evaluated under two approaches: (1) with recovery rate and cost per child admitted for treatment as measures of effectiveness and cost, respectively; and (2) assessing the total number of children recovered and the total cost incurred. In addition, a multivariate probabilistic sensitivity analysis was carried out to evaluate the effect of uncertainty around the base case input data. RESULTS: For the base case data, the average cost per child recovered was 116.52 USD in the standard treatment and 107.22 USD in the CHWs-led treatment. Based on the first approach, the CHWs-led treatment was more cost-effective than the standard treatment with an average cost per child admitted for treatment of 82.81 USD vs. 84.01 USD. Based on the second approach, the incremental cost-effectiveness ratio of the transition from the standard to the CHWs-led treatment amounted to 98.01 USD per additional SAM case recovered. CONCLUSIONS: In the district of Mayahi in Niger, the CHWs-led SAM treatment was found to be cost-effective when compared to the standard protocol and provided additional advantages such as the reduction of costs for households. TRIAL REGISTRATION: ISRCTN with ID 31143316. https://doi.org/10.1186/ISRCTN31143316.

Qi-dan-dihuang decoction ameliorates renal fibrosis in diabetic rats via p38MAPK/AKT/mTOR signaling pathway.

CONTEXT: Qi-dan-dihuang decoction (QDD) has been used to treat diabetic kidney disease (DKD), but the underlying mechanisms are poorly understood. OBJECTIVE: This study reveals the mechanism by which QDD ameliorates DKD. MATERIALS AND METHODS: The compounds in QDD were identified by high-performance liquid chromatography and quadrupole-time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS). Key targets and signaling pathways were screened through bioinformatics. Nondiabetic Lepr db/m mice were used as control group, while Lepr db/db mice were divided into model group, dapagliflozin group, 1% QDD-low (QDD-L), and 2% QDD-high (QDD-H) group. After 12 weeks of administration, 24 h urinary protein, serum creatinine, and blood urea nitrogen levels were detected. Kidney tissues damage and fibrosis were evaluated by pathological staining. In addition, 30 mmol/L glucose-treated HK-2 and NRK-52E cells to induce DKD model. Cell activity and migration capacity as well as protein expression levels were evaluated. RESULTS: A total of 46 key target genes were identified. Functional enrichment analyses showed that key target genes were significantly enriched in the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) and mitogen-activated protein kinase (MAPK) signaling pathways. In addition, in vivo and in vitro experiments confirmed that QDD ameliorated renal fibrosis in diabetic mice by resolving inflammation and inhibiting the epithelial-mesenchymal transition (EMT) via the p38MAPK and AKT-mammalian target of rapamycin (mTOR) pathways. DISCUSSION AND CONCLUSION: QDD inhibits EMT and the inflammatory response through the p38MAPK and AKT/mTOR signaling pathways, thereby playing a protective role in renal fibrosis in DKD.

A Comprehensive Analysis of Fel Ursi and Its Common Adulterants Based on UHPLC-QTOF-MSE and Chemometrics.

BACKGROUND: As one of the four most valuable animal medicines, Fel Ursi, named Xiong Dan (XD) in China, has the effect of clearing heat, calming the liver, and brightening the eyes. However, due to the special source of XD and its high price, other animals' bile is often sold as XD or mixed with XD on the market, seriously affecting its clinical efficacy and consumers' rights and interests. In order to realize identification and adulteration analysis of XD, UHPLC-QTOF-MSE and multivariate statistical analysis were used to explore the differences in XD and six other animals' bile. METHODS: XD, pig gall (Zhu Dan, ZD), cow gall (Niu Dan, ND), rabbit gallbladder (Tu Dan, TD), duck gall (Yan Dan, YD), sheep gall (Yang Dan, YND), and chicken gall (Ji Dan, JD) were analyzed by UHPLC-QTOF-MSE, and the MS data, combined with multivariate analysis methods, were used to distinguish between them. Meanwhile, the potential chemical composition markers that contribute to their differences were further explored. RESULTS: The results showed that XD and six other animals' bile can be distinguished from each other obviously, with 27 ions with VIP > 1.0. We preliminarily identified 10 different bile acid-like components in XD and the other animals' bile with significant differences (p < 0.01) and VIP > 1.0, such as tauroursodeoxycholic acid, Glycohyodeoxycholic acid, and Glycodeoxycholic acid. CONCLUSIONS: The developed method was efficient and rapid in accurately distinguishing between XD and six other animals' bile. Based on the obtained chemical composition markers, it is beneficial to strengthen quality control for bile medicines.

Clinical Study on the Emotional Intervention of Patients with Asymptomatic and Mild Novel Coronavirus (COVID-19).

Background and Objectives: The aim was to explore the interventional effect of the traditional Chinese medicine (TCM) exercise of Tian Dan Shugan Tiaoxi on the emotions of patients with mild novel coronavirus (COVID-19). Materials and Methods: A total of 110 asymptomatic and mildly symptomatic COVID-19 patients from Hongkou Memorial Road Temporary Cabin Hospital and South Renji Hospital were selected between April 2022 and June 2022, and randomly divided into two groups: a control group and an intervention group. There were 55 participants in each group. The control group was treated with Lianhua Qingwen granules, and members of the intervention group were made to practice Tian Dan Shugan Tiaoxi (an exercise that soothes the liver and regulates emotions) every day for 5 days. The Patient Health Questionnaire-9 (PHQ-9), the Generalized Anxiety Disorder questionnaire (GAD-7), and the Symptom Checklist 90 (SCL-90) were used to evaluate the data collected before and after the trial. Results: The incidence of anxiety and depression was high in the patients included in this study, at 73.64% and 69.09%, respectively. After intervention, the scores of the Patient Health Questionnaire-9 (PHQ-9) and the Generalized Anxiety Disorder questionnaire (GAD-7) in the two groups had decreased in comparison with those recorded before intervention (p < 0.05). The PHQ-9 and GAD-7 scores in the intervention group were significantly better than those of the control group (p < 0.05). The factors of somatization, depression, anxiety, hostility, and fear in the SCL-90 in the intervention group were significantly improved after intervention, and generally, better than those in the control group (p < 0.05). Conclusions: Patients infected with novel coronavirus in shelter hospitals have different degrees of emotional abnormalities. Tian Dan Shugan Tiaoxi can reduce the anxiety and depression of people with mild novel coronavirus, and it can be practiced clinically to improve the recovery rate among infected people.

[Preparation of Huoluo Xiaoling gel plaster and its transdermal penetration in vitro].

Huoluo Xiaoling Dan is a classical prescription commonly used for blood circulation and pain relief in clinic with obvious effects. To make it directly treat lesion and improve the effect, this research optimized the preparation process of Huoluo Xiaoling gel paste and further evaluated its in vitro transdermal absorption performance, so as to provide a scientific basis for its development and utilization. Using primary viscosity, holding viscosity, and sensory score as evaluation indexes, the matrix amount of gel paste was determined by the single factor test and Box-Behnken response surface method. The ultra-performance liquid chromatography(UPLC) method was established to determine the content of eight active ingredients, including Danshensu, ferulic acid, salvianolic acid B, salvianolic acid A, ligustilide, tanshinone Ⅱ_A, 11-keto-ß-boswellic(KBA), and 3-acetyl-11-keto-ß-boswellic acid(AKBA). A mo-dified Franz diffusion cell method was used to evaluate and compare the absorption properties of the gel paste without volatile oil and with volatile oil microemulsion. The results showed that the optimal prescription for Huoluo Xiaoling gel paste matrix was NP700(1.35 g), glycerol(7.00 g), micropowder silica gel(1.25 g), sodium carboxymethyl cellulose(0.20 g), tartaric acid(0.06 g), and glyceryl aluminum(0.04 g). The mass fractions of eight active ingredients in the paste were successively 0.48, 0.014, 0.95, 0.39, 0.57, 0.055, 0.35, and 0.97 mg·g~(-1). The results of the in vitro transdermal absorption test showed that the addition of the volatile oil or the volatile oil microemulsion promoted the transdermal absorption of the active ingredients, and the law of drug penetration conformed to the zero equation or the Higuchi equation. The gel paste prepared by the optimal prescription has good appearance and adhesion, with no residue, and has the characteristics of skeletal slow-release preparation, which is easy to reduce the number of administration, la-ying a foundation for the development of new external dosage forms of Huoluo Xiaoling Dan.

Lipidomic-based investigation into the therapeutic effects of polyene phosphatidylcholine and Babao Dan on rats with non-alcoholic fatty liver disease.

In recent years, with the improvement of people's living standards, non-alcoholic fatty liver disease (NAFLD) has become the most common chronic liver disease in the world. In this paper, the metabolic disorders in Sprague Dawley (SD) rats were induced by a choline-deficient, l-amino acid-defined (CDAA) diet. The therapeutic effects of polyene phosphatidylcholine (PPC) and Babao Dan (BBD) on NAFLD were observed. Lipidomic analysis was performed using ultra-high-performance liquid chromatography-Orbitrap MS, and data analysis and lipid identification were performed using the software LipidSearch. Both PPC and BBD can reduce lipid accumulation in the liver and improve abnormal biochemical indicators in rats, including reduction of triglycerides, total cholesterol, alanine transaminase and aspartate transaminase in serum. In addition, lipids in rat serum were systematically analyzed by lipidomics. The lipidomic results showed that the most obvious lipids with abnormal metabolism in CDAA diet-induced rats were glycerides (triglycerides and diacylglycerols), phospholipids and cholesterol esters. Both BBD and PPC partly reversed the disturbance to lipids induced by the CDAA diet. PPC may be more effective than BBD in alleviating NAFLD because it has a better effect on inhibiting the abnormal accumulation of lipids and reducing the inflammatory reaction in the body.

In vivo and in vitro metabolism study of traditional Chinese medicine formula Dingkun Dan in rats by using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

Dingkun Dan (DKD), a reputable traditional Chinese medicine formula, has been used to treat gynecological diseases and showed significant clinical effects since ancient times. However, the application and development of DKD are seriously hampered by the unclear active substances. Structural characterization of compounds absorbed in vivo and their corresponding metabolites is significant for clarifying the pharmacodynamic material basis. In this study, an integrated strategy using ultra-performance liquid chromatography, coupled with quadrupole time-of-flight mass spectrometry and UNIFI™ software, was used to identify prototypes and metabolites after oral administration of DKD in rats. As a result, a total of 261 compounds, including 140 prototypes and 121 metabolites, were tentatively characterized in rat plasma, urine, and feces. The metabolic pathways of prototypes have been studied to clarify their possible transformation process in vivo. Moreover, an in vitro metabolism study was applied for verifying the metabolites under simulating the metabolic environment in vivo. This first systematic metabolic study of DKD is important for elucidating the metabolites and metabolic pathways and could provide a scientific basis for explaining the integrative mechanism in further pharmacology study.

[Action mechanism of Professor Xu Fusong's Shuilu Erxian Dan in the treatment of premature ejaculation: An exploration based on network pharmacology and molecular docking technology].

OBJECTIVE: To analyze the potential action mechanism of Shuilu Erxian Dan in the treatment of premature ejaculation based on network pharmacology. METHODS: The TCMSP database was searched for the main chemical constituents of Euryale and golden cherry, the core components of Shuilu Erxian Dan. Oral bioavailability and medicinal properties were set as the parameters for screening active components, premature ejaculation-related disease targets retrieved from the DisGeNET, GeneCards, OMIM and TTD databases, the common targets of drug pairs and diseases identified with the Wayne diagram, and a drug-active ingredient-target-disease network map constructed with the software Cytoscape. A PPI network map designed through the STRING platform, the website Metascape used for GO and KEGG analyses of the common targets, and the protein receptors and small molecular ligands of the core genes employed for molecular docking and PyMOL visualization. RESULTS: A total of 7 chemical constituents and 171 predictive targets, 1 918 premature ejaculation targets and 113 common targets were identified. GO analysis showed significant enrichment of BP in response to toxic substances and inorganic substances, CC in membrane raft and plasma membrane protein complex, and MF in transcription factor binding and kinase binding. KEGG analysis manifested markedly enriched TNF signaling pathway, JAK-STAT signaling pathway and Cocaine addiction. The results of molecular docking exhibited a good binding energy between the core gene and the component. CONCLUSION: Network pharmacology has revealed the potential action mechanism of Shuilu Erxian Dan in the treatment of premature ejaculation, which has paved the theoretical ground for verification of the effectiveness of Shuilu Erxian Dan in the treatment of premature ejaculation via animals and cells.

Asymmetric faces: Symbolic, spiritual, and representative.

In humans, physically attractive faces are measurably, though subtly, asymmetric. As asymmetry increases, it has a negative impact. Medically, asymmetry can be congenital or acquired. Symbolically, it has varied connotations from playfulness and complexity to despair and corruption. In Chinese opera, stylized make-up exaggerates the features, and aspects of the "mask" indicate qualities of the character. The asymmetric faces belong to characters who are corrupt, devious, or evil. In the Dan culture of western Africa, performance masks channel spirits in the community. A spirit with asymmetric facial mask exemplifies ugliness and moral failing. The Nasca culture of South America made generic figures of farmers, deities, and so on, but not of individuals. However, there is evidence of mutual influence between the Nasca and the Wari, with whom they traded. A clay figure apparently representing an individual, or at least a very specific recognized persona, is a ball player with facial asymmetry presumably due to injury. Here the message is one of fierceness and strength. The relative rarity of asymmetric facial depictions compared to symmetric ones is cross-cultural. This implies that asymmetry is special somehow, in all connotations of that term.

Examination of mercury contamination from a recent coal ash spill into the Dan River, North Carolina, United States.

Coal ash spills occasionally occur due to the accidental failure of surface impoundments, and toxic metal-laden ash can pose a serious health threat to adjacent aquatic ecosystems. Here, we performed an investigation into longitudinal variations of mercury (Hg) contamination in the Dan River (North Carolina, United States) about 17 and 29 months after a February 2014 coal ash spill incident, in which the reported Hg concentrations in the spilled coal ash (210 ng/g) were 1-2 orders of magnitude higher than the river sediments (2-61 ng/g). We examined total Hg (THg) and methyl Hg (MeHg) in sediments from 0 to 65 km downstream of the spill, and found that most of the variations of THg and MeHg in surface sediments (0-16 cm) could be well accounted by the organic matter content and appeared to be not contaminated by Hg derived from coal ash. In examining MeHg bioaccumulation in invertebrates (aquatic and riparian) and fish in the Dan River and fish in a reservoir downstream of Dan River, we found no evidence of elevated MeHg bioaccumulation due to the 2014 coal ash spill. Thus, we concluded that Hg contamination from the coal ash spill is largely absent in the Dan River for both surface sediments and biota within the first three years of spill (until 2017), even though the majority of coal ash may be buried deeper in the sediment in the river channel and/or the downstream reservoir. Alternatively, the Hg associated with the coal ash is largely not bioavailable for extensive microbial Hg methylation. The findings provide useful insights into remediation strategies for this incident and other coal ash spills.

Charge-Transfer Interactions Stabilize G-Quadruplex-Forming Thrombin Binding Aptamers and Can Improve Their Anticoagulant Activity.

In the search for optimized thrombin binding aptamers (TBAs), we herein describe the synthesis of a library of TBA analogues obtained by end-functionalization with the electron-rich 1,5-dialkoxy naphthalene (DAN) and the electron-deficient 1,8,4,5-naphthalenetetra-carboxylic diimide (NDI) moieties. Indeed, when these G-rich oligonucleotides were folded into the peculiar TBA G-quadruplex (G4) structure, effective donor-acceptor charge transfer interactions between the DAN and NDI residues attached to the extremities of the sequence were induced, providing pseudo-cyclic structures. Alternatively, insertion of NDI groups at both extremities produced TBA analogues stabilized by π-π stacking interactions. All the doubly-modified TBAs were characterized by different biophysical techniques and compared with the analogues carrying only the DAN or NDI residue and unmodified TBA. These modified TBAs exhibited higher nuclease resistance, and their G4 structures were markedly stabilized, as evidenced by increased Tm values compared to TBA. These favorable properties were also associated with improved anticoagulant activity for one DAN/NDI-modified TBA, and for one NDI/NDI-modified TBA. Our results indicated that TBA pseudo-cyclic structuring by ad hoc designed end-functionalization represents an efficient approach to improve the aptamer features, while pre-organizing and stabilizing the G4 structure but allowing sufficient flexibility to the aptamer folding, which is necessary for optimal thrombin recognition.

[Influence of Dingkun Dan Combined with Estradiol Valerateon Wnt/ß-catenin Signaling Pathway in Rats with Thin Endometrium with Kidney-Yang Deficiency].

OBJECTIVE: To explore the influence for combination of Dingkun Dan with estradiol valerateon in treating rats with thin endometrium with Kidney-Yang deficiency based on Wnt/ß-catenin signaling pathway. METHODS: The estrous period 40 rats were randomly divided into the normal control group, the model group, the Dingkun Dan group, the estradiol valerateon group and the combination group, with 8 rats in each group. In addition to the normal control group, the rat model of thin endometrium with Kidney-Yang deficiency was established in other groups. The control group used free diet, the model group was given distilled water, the estradiol valerateon group was treated with progynova by gavage at 0.3 mg/(kg·d) , Dingkun Dan group was treated with Dingkun Dan by gavage at 2.26 g/(kg·d), and the combined group was given Dingkun Dan at 2.26 g/(kg·d) on the basis of progynova at 0.3 mg/(kg·d). After 3 estrous cycles, the rats were killed and harvested. HE staining was used to observe histopathologic changes in endometrium. The expression of VEGF in rats endometrium were detected by immunohistochemistry. The expression of ß-catenin, E-cadherin and MMP-9 protein in rat endometrium was detected by Western blot. RESULTS: Compared with the control group, the uterine cavity was narrowed or enlarged, the endometrium glands and blood vessels were sparse, and the endometrium was thinner significantly in the model group ( P<0.01); the levels of VEGF was decreased significantly ( P<0.01), while ß-catenin, E-cadherin and MMP-9 were increased significantly ( P<0.01). Compared with the model group, more endometrial glands, rich intimal vessels, the endometrium were thickened significantly in the 3 treatment groups ( P<0.01 or P<0.05); the levels of VEGF was increased differently. The protein levels of ß-catenin and E-cadherin were significantly decreased in each treatment group ( P<0.01), and MMP-9 were significantly decreased in the Dingkun Dan group and in the combination group ( P<0.01). Compared with the estradiol valerateon group, the level of ß-catenin in Dingkun Dan group was higher, and MMP-9 was lower ( P<0.05 or P<0.01). The levels of ß-catenin and MMP-9 in the combination group were significantly decreased ( P<0.01). Compared with the combination group, the levels of ß-catenin was increased significantly, while decreased signicantly in Dingkun Dan group ( P<0.01 or P<0.05). CONCLUSION: Dingkun Dan combined with estradiol valerateon can increase the thicken of the endometrium by up-regulation of VEGF, while down-regulate of ß-catenin, E-cadherin and MMP-9 in rats with Shen-Yang deficiency and thin endometrium.

COVID: Using the DAN Model with Couples to Increase Family Resilience.

The COVID-19 pandemic has massively impacted our society, affecting both wider systems and smaller ones alike. Families across the world have had to adapt to a new style of living, dominated by uncertainty, which is testing their resilience. Within this context, we have asked ourselves, as systemic therapists, what can be done to provide support and a sense of understanding to families. In this article, it is argued that therapeutic tools taken from a manualised intervention to promote resilience in couples can provide valuable help to families who are struggling in this difficult time. This intervention, developed at the Iscra Institute in Modena, Italy, is called Digital, Analogic, and Narrative (DAN). The tools used in the DAN model are described in this paper and their application in practice is illustrated through a case study.

[Pharmacokinetics of eight active ingredients of Salviae Miltiorrhizae Radix et Rhizoma-Puerariae Lobatae Radix combination in rats].

To reveal the rationality of compatibility of Salviae Miltiorrhizae Radix et Rhizoma(SMRR) and Puerariae Lobatae Radix(PLR) from the perspective of pharmacokinetics, this study established a UPLC-MS/MS method for quantitative determination of PLR flavonoids(3'-hydroxy puerarin, puerarin, puerarin 6″-O-xyloside, 3'-methoxy puerarin, puerarin apioside) and salvianolic acids and tanshinones(salvianolic acid B, cryptotanshinone, and tanshinone Ⅱ_A) in plasma of rats. Rats were given SMRR extract, PLR extract, and SMRR-PLR extract by gavage and then plasma was collected at different time. UPLC separation was performed under the following conditions: Eclipse C_(18) column(2.1 mm×50 mm, 1.8 µm), 0.1% formic acid in water(A)-0.1% formic acid in acetonitrile(B) as mobile phase for gradient elution. Conditions for MS are as below: multiple reaction monitoring(MRM), ESI~(+/-). Comprehensive validation of the UPLC-MS/MS method(specifically, from the aspects of calibration curve, precision, accuracy, repeatability, stability, matrix effect, extract recovery) was performed and the result demonstrated that it complied with quantitative analysis requirements for biological samples. Compared with SMRR extract alone or PLR extract alone, SMRR-PLR extract significantly increased the AUC and C_(max) of PLR flavonoids and tanshinones in rat plasma, suggesting that the combination of SMRR and PLR promoted the absorption of the above components. The underlying mechanism needs to be further studied.

Microbial community succession and toxic alkaloids change during fermentation of Huafeng Dan Yaomu.

Fermentation is a traditional processing method that can impact the abundance of relevant components in Chinese herbal medicines. Huafeng Dan Yaomu was processed by fermentation. In this study, we investigated changes in toxic alkaloids using high-performance liquid chromatography and analysed the changes in microbial communities during fermentation of Huafeng Dan Yaomu by Illumina MiSeq platform. The results indicated that highly toxic alkaloid compounds including aconitine, mesaconitine and hypaconitine were decreased, whereas benzoylmesaconine and benzoylhypaconitine were increased during fermentation. The dominant bacterial genera in the raw material (day 0) were Bacillus (58·38%), Enterobacter (16·05%), Enterococcus (4·10%) and others (11·48%). After 7 days of fermentation, Pediococcus predominated, increasing from 98·26 to 99·97%. The dominant fungal genera in the raw material (day 0) were Incertae_Sedis_incertae_sedis (45·36%), Eurotiales_unclassified (20·63%), Millerozyma (15·58%) and Saccharomycopsis (10·64%). After 7 days of fermentation, Saccharomycopsis was increased to 90·84%. After 14 days of fermentation, Pichia became the main fungal genera in the fermentation process. There were 16 bacterial genera and seven fungal genera that displayed significant correlation with toxic alkaloids. The dynamics of the microbiota and chemical compounds during fermentation of Huafeng Dan Yaomu were revealed, providing a foundation for further investigation of the effects of microbes on chemical compounds and insights into the manufacturing of Huafeng Dan Yaomu. SIGNIFICANCE AND IMPACT OF THE STUDY: Huafeng Dan Yaomu is a unique fermentation-based micro-organism product. This study reveals that the change in toxic alkaloids during fermentation was related to micro-organisms. The results of this study can be used to improve the Chinese herb medicine fermentation process, such as the selection of beneficial strains and their composition for modern fermentation. The results of this study may also provide new ideas for the processing of Traditional Chinese Medicine.

Mapping cognitive and emotional networks in neurosurgical patients using resting-state functional magnetic resonance imaging.

Neurosurgery has been at the forefront of a paradigm shift from a localizationist perspective to a network-based approach to brain mapping. Over the last 2 decades, we have seen dramatic improvements in the way we can image the human brain and noninvasively estimate the location of critical functional networks. In certain patients with brain tumors and epilepsy, intraoperative electrical stimulation has revealed direct links between these networks and their function. The focus of these techniques has rightfully been identification and preservation of so-called "eloquent" brain functions (i.e., motor and language), but there is building momentum for more extensive mapping of cognitive and emotional networks. In addition, there is growing interest in mapping these functions in patients with a broad range of neurosurgical diseases. Resting-state functional MRI (rs-fMRI) is a noninvasive imaging modality that is able to measure spontaneous low-frequency blood oxygen level-dependent signal fluctuations at rest to infer neuronal activity. Rs-fMRI may be able to map cognitive and emotional networks for individual patients. In this review, the authors give an overview of the rs-fMRI technique and associated cognitive and emotional resting-state networks, discuss the potential applications of rs-fMRI, and propose future directions for the mapping of cognition and emotion in neurosurgical patients.

[Effects of Dingkun Dan combined with Western medicine on endometrial ß-catenin, VEGF and MMP-9 mRNA expression in rats with multiple lesions].

The aim of this paper was to investigate the effect of Dingkun Dan on endometrial receptivity in rats with multiple lesions. Forty SD female rats with regular sexual cycle were randomly divided into 5 groups, control group, model group, progynova group, Dingkun Dan group and combination group. The thin endometrium model of kidney-yang deficiency was established in all the other rats except normal control group. The rats in normal control group were free to drink and eat; the rats in the model group were administered with distilled water; the rats in the progynova group were treated with progynova; rats in Dingkun Dan group were treated with Dingkun Dan, and the rats in combination group were treated with Dingkun Dan and progynova. After 15 days, serum levels of OPN, VEGF and MMP-9 were measured by ELISA. HE staining, immunohistochemistry and RT-PCR were used to analyze endome-trial morphology, endometrial thickness and the treatment mechanism of Dingkun Dan. As compared with the control group, the serum levels of OPN, VEGF and MMP-9 in the model group were significantly increased(P<0.01). As compared with the model group, the serum levels of OPN and MMP-9 were decreased in Dingkun Dan group(P<0.05, P<0.01). As compared with the control group, endometrial stromal cells were fewer, the endometrium glands and blood vessels were sparse, and the endometrium was thinner significantly in the model group(P<0.01). As compared with the model group, there were more endometrial glands, rich intimal vessels, and dense stromal cells in various treatment groups, and the endometrium were thickened significantly in the treatment groups(P<0.01). As compared with the control group, the expression area of CK19 in the model group was decreased significantly(P<0.01). As compared with the model group, the expression area of CK19 in each treatment group was increased significantly(P<0.05). As compared with the control group, endometrial ß-catenin and MMP-9 mRNA expression levels were increased significantly in the model group(P<0.05), while VEGF mRNA expression was decreased(P<0.05). As compared with the model group, MMP-9 mRNA expression was decreased significantly in the progynova group and the combination group(P<0.05). Dingkun Dan combined with progynova can improve endometrial receptivity by up-regulating expression of ß-catenin, VEGF mRNA and down-regulating the expression of MMP-9 mRNA in the injury rats.