A B-box transcription factor OsBBX17 regulates saline-alkaline tolerance through the MAPK cascade pathway in rice.

Rice OsBBX17 encodes a B-box zinc finger transcription factor in which the N-terminal B-box structural domain interacts with OsMPK1. In addition, it directly binds to the G-box of OsHAK2 and OsHAK7 promoters and represses their transcription. Under saline-alkaline conditions, the expression of OsBBX17 was inhibited. Meanwhile, activation of the OsMPK1-mediated mitogen-activated protein kinase cascade pathway caused OsMPK1 to interact with OsBBX17 and phosphorylate OsBBX17 at the Thr-95 site. It reduced OsBBX17 DNA-binding activity and enhanced saline-alkaline tolerance by deregulating transcriptional repression of OsHAK2 and OsHAK7. Genetic assays showed that the osbbx17-KO had an excellent saline-alkaline tolerance, whereas the opposite was in OsBBX17-OE. In addition, overexpression of OsMPK1 significantly improved saline-alkaline tolerance, but knockout of OsMPK1 caused an increased sensitivity. Further overexpression of OsBBX17 in the osmpk1-KO caused extreme saline-alkaline sensitivity, even a quick death. OsBBX17 was validated in saline-alkaline tolerance from two independent aspects, transcriptional level and post-translational protein modification, unveiling a mechanistic framework by which OsMPK1-mediated phosphorylation of OsBBX17 regulates the transcription of OsHAK2 and OsHAK7 to enhance the Na+ /K+ homeostasis, which partially explains light on the molecular mechanisms of rice responds to saline-alkaline stress via B-box transcription factors for the genetic engineering of saline-alkaline tolerant crops.

Metabolite profiles of epimedin C in rat plasma and bile by ultra-performance liquid chromatography coupled with quadrupole-TOF-MS.

Epimedin C is one of the major bioactive constituents of Herba Epimedii. In this study, the metabolite profiles of epimedin C in rat plasma and bile were qualitatively investigated, and the possible metabolic pathways of epimedin C were subsequently proposed. After oral administration of epimedin C at a single dose of 80 mg/kg, rat biological samples were collected and pretreated by protein precipitation. Then these pretreated samples were injected into an Acquity UPLC BEH C18 column and detected by ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry. In all, 12 metabolites were identified in the biosamples. Of these, eight, two from plasma and six from bile, are, to our knowledge, reported here for the first time. The results indicated that epimedin C was metabolized via desugarization, dehydrogenation, hydrogenation, dehydroxylation, hydroxylation, demethylation and glucuronidation pathways in vivo. Thus, this study revealed the possible metabolite profiles of epimedin C in rat plasma and bile.

[Two-dimensional zebrafish model combined with hyphenated chromatographic techniques for evaluation anti-osteoporosis activity of epimendin A and its metabolite baohuoside I].

This article firstly established a new efficient method for screening anti-osteoporosis ingredients, which used two-dimensional zebrafish model combined with hyphenated chromatographic techniques to evaluate anti-osteoporosis activities of epimedin A and its metabolite baohuoside I. Adult zebrafish was used for metabolism of epimedin A in 0.5% DMSO, and LC-MS was used for analysis of the metabolite, which was captured by HPLC, and prednisolone-induced osteoporosis model of zebrafish was used to evaluate the anti-osteoporotic activities of trace amounts of epimedin A and baohuoside I. The results indicated that epimedin A and baohuoside I can prevent prednisolone-induced osteoporosis in zebrafish. The developed method in this paper enables the separation, enrichment and analysis of micro-amount metabolite of epimedin A, and anti-osteoporosis activities in vivo of epimedin A and baohuoside I was simple and efficient screening resorting to zebrafish osteoporosis mode. This paper would provide new ideas and methods for a rapid and early discovery of anti-osteoporosis activities of micro-ingredients and its metabolite of traditional Chinese medicine.

[Discussion about research progress and ideas on processing mechanism of traditional Chinese medicine].

Study on the processing mechanism of traditional Chinese medicine is the key to science of processing Chinese materia medica and modernization of traditional Chinese medicine. At present, chemical and pharmacology methods are mainly used to discuss the processing principle of efficiency, attenuated, delayed or cooked with different treatment. So that the processing mechanism of Chinese herbal medicine has made breakthrough progress. With the introduction of modern science and technology, biotransformation, intestinal absorption, pharmacokinetics and metabolomics methods have been gradually applied in traditional Chinese medicine processing mechanism. This article summarizes the achievements in the processing mechanism of traditional Chinese medicine in recent years, analyses and discusses some main problems, and points out to in-depth study on absorption and metabolism, strengthening excipient processing mechanism, paying attention to the integration of multiple disciplines and data statistical analysis. Combined with years of exploration and practice, the project group proposes a new idea "traditional Chinese medicine processing mechanism based on coupled effect of chemical composition transformation and intestinal absorption barrier" , which provides reference for the study of the mechanism of traditional Chinese medicine processing.

[Study on preparation of sagittatoside B with epimedin B converted from cellulase].

To prepare sagittatoside B with epimedin B Hydrolyzed from cellulase. With the conversion ratio as the index, the effects of pH value, temperature, reaction time, dosage of enzyme and concentration of substrates on the conversion ratio were detected. L9 (3(4)) orthogonal design was adopted to optimize the preparation process. Hydrolyzed products were identified by MS, 1H-NMR, and 13C-NMR. The results showed that the optimum reaction conditions for the enzymatic hydrolysis were that the temperature was 50 degrees C, the reaction medium was pH 5.6 acetic acid-sodium acetate buffer solution, the concentration of substrates was 20 g x L(-1), the mass ratio between enzyme and substrate was 3: 5, and the relative molecular mass of the reaction product was 646.23. NMR data proved that the product was sagittatoside B. The process is simple and reliable under mild reaction conditions, thus suitable for industrial production.

[Effect of suet oil on formation of self-assembled micelles of flavonoids in Epimedium].

In this paper, the action of suet oil in the preparation of self-assembled micelles of the active flavonoids in Epimedium in the simulated human environment was researched. Twelve suet oil samples were collected from different growing areas and different positions of sheep or goat to simulate the formation of micelles. Then the effects of the fatty acids in suet oil on the preparation of self-assembled micelles were studied furthermore. The results showed that the micelles had a dispersed state and spherical smooth surface. To compare the diameter, potential, encapsulation efficiency and drug loading of the 12 batches micelles, the micelles prepared by the suet oil from Qinghai were more stable and had a higher encapsulation efficiency. The fatty acids in suet oil could promote the formation of self-assembled micelles, but the whole suet oil had a better effect. Above all the study, we confirmed that the suet oil promoted the formation of self-assembled micelles of the flavonoids in Epimedium, it laid foundation for further research about increasing the efficacy of Epimedium and improved the absorption of the active flavonoids in Epimedium.

[Discussion on research ideas of synergistic mechanism of Epimedium fried with suet oil based on self-assembled micelles formation in vivo].

Accessories processing is a major feature of traditional pharmaceutical technology in China How to use modem science and technology to reveal the scientific connotation of accessories processing, is the core of traditional Chinese medicine processing research. As traditional beneficial Chinese medicine, Epimedium can enhance the effect of warming kidney yang after processing with suet oil However, the current literature research from two aspects of chemical constituents and pharmacological action cannot fully explain the processing mechanism of accessories suet oil. According to the characteristics of fatty acids in suet oil with long chain fatty and surface activity, and based on the theoretical basis of self-assembled drug delivery system, as well as the experimental basis of Epimedium flavonoids self-assembled micelles simulated formation and promoting absorption, this paper puts forward the research hypothesis on synergistic mechanism of Epimedium fried with suet oil based on self-assembled micelles formation in vivo. This paper also describes in detail the research ideas and methods of self-assembled micelles enhancing absorption and synergistic mechanism of Epimedium flavonoids, which has the important theory significance and application value to reveal the scientific connotation of Chinese medicine fried with oil.

[Processing mechanism of Epimedium fried with suet oil based on absorption and metabolism of flavonoids].

As beneficial traditional Chinese medicine, Epimedium fried with suet oil can enhance the effect of warming kidney yang. Previous literature studies about processing of Epimedium mainly focused on changes in chemical composition and pharmacological effect. From the angle of flavonoids absorption and metabolism, our group innovatively study the processing mechanism of Epimedium based on active component transformation combined with intestinal absorption barrier. The processing effect of fried Epimedium can be divided into two key aspects of " heat" during processing and processing accessories "suet oil". Through continuous three National Natural Science Foundation items, the research group has clarified the scientific connotation of "heat" during processing with ADME, and explains the synergistic mechanism of processing accessories "suet oil" based on self-assembled micelles formation in vivo for the first time. This paper summarizes the research ideas and results of Epimedium processing mechanism of the project team for many years, and discusses the future research direction and content, in order to provide new ideas and new methods for modern Chinese medicine processing mechanism.

[Construction of research system for processing mechanism of traditional Chinese medicine based on chemical composition transformation combined with intestinal absorption barrier].

Based on practice of Epimedium processing mechanism for many years and integrated multidisciplinary theory and technology, this paper initially constructs the research system for processing mechanism of traditional Chinese medicine based on chemical composition transformation combined with intestinal absorption barrier, which to form an innovative research mode of the " chemical composition changes-biological transformation-metabolism in vitro and in vivo-intestinal absorption-pharmacokinetic combined pharmacodynamic-pharmacodynamic mechanism". Combined with specific examples of Epimedium and other Chinese herbal medicine processing mechanism, this paper also discusses the academic thoughts, research methods and key technologies of this research system, which will be conducive to systematically reveal the modem scientific connotation of traditional Chinese medicine processing, and enrich the theory of Chinese herbal medicine processing.

Synthesis of 3,3-disubstituted oxindoles by visible-light-mediated radical reactions of aryl diazonium salts with N-arylacrylamides.

A mild and efficient visible-light-mediated diarylation of N-arylacrylamides with aryl diazonium salts under mild conditions has been developed. This method provides convenient access to a variety of useful 3,3-disubstituted oxindoles by constructing two C-C bonds in one step.

[Simultaneous content determination of notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and dracorhodin in ZJHX rubber paste by double wavelength HPLC].

Notoginsenoside R1, ginsenoside Rg1, Re, Rb, and dracorhodin from ZJHX rubber paste were analyzed simultaneously by HPLC, with acetonitrile-water as the mobile phase for gradient elution at a flow rate of 1.0 mL x min(-1). The column temperature was 35 degrees C and the sample size was 10 microL. The detection wavelength was set at 203 nm for ginsenoside and 440 nm for dracorhodin, respectively. The results showed that all of notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and dracorhodin could be were separated well by baseline, with the linear ranges of 0.251-5.020 microg (R2 = 0.999 8), 0.520-10.400 microg (R2 = 0.999 9), 0.251-5.010 microg (R2 = 0.999 7), 0.505-10.100 microg (R2 = 0.999 8) and 0.160-3.270 microg (R2 = 0.999 9), respectively. Each component showed a good linear relationship, with the average recoveries ranging from 99.39% to 100.5%. The established method was so simple, accurate and highly reproducible that it could be used for quality control of ZJHX rubber paste.

The UDP-glucuronic acid decarboxylase OsUXS3 regulates Na+ ion toxicity tolerance under salt stress by interacting with OsCATs in rice.

Overly Na+ ion in soil caused by salt stress has a significant negative impact on the growth and production of crops, especially rice (Oryza sativa L.). Therefore, it is vital for us to clarify how salt stress tolerance in rice is caused by Na+ ion toxicity. The UDP-glucuronic acid decarboxylase (UXS) is a critical enzyme in the biosynthesis of UDP-xylose, which is the key substrate of cytoderm synthesis in plants. In this study, we found that OsUXS3, a rice UXS, is a positive regulator to regulate Na+ ion toxicity under salt stress by interacting with OsCATs (Oryza sativa catalase; OsCAT). The expression of OsUXS3 was significantly up-regulated under NaCl and NaHCO3 treatments of rice seedlings. Meanwhile, by the genetic and biochemical evidence, knockout of OsUXS3 significantly increased reactive oxygen species (ROS) levels and decreased CAT activity under NaCl and NaHCO3 treatments in tissue. Furthermore, knockout of OsUXS3 caused excessive accumulation of Na + ion and rapid loss of K+ ion and disrupts Na+/K+ homeostasis under NaCl and NaHCO3 treatments. Based on the results above, we can conclude that OsUXS3 might regulate CAT activity by interacting with OsCATs, which is not only characterized for the first time but also regulating Na+/K+ homeostasis, positively regulating the Na+ ion toxicity tolerance under salt stress in rice.

Characterization of metabolites of sagittatoside B in rats using UPLC-QTOF-MS spectrometry.

Sagittatoside B is one of the principal diglucosides in Herba Epimedii. In this work, an ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) was applied to the rapid analysis of sagittatoside B metabolites in rats after oral administration. A total number of 17 metabolites were detected or tentatively identified from rat plasma, bile, urine and feces. The major metabolic pathways of sagittatoside B in rats were hydrolysis, hydrogenation, hydroxylation, dehydrogenation, demethylation, decarbonylation and conjugation with glucuronic acid and different sugars. This work revealed the metabolism of sagittatoside B in vivo, and reported the characteristic metabolic reactions of sagittatoside B for the first time. This provided the basis for the further research and development of sagittatoside B, and also provided reference for the metabolism of active flavonoid compounds.

ECA-TFUnet: A U-shaped CNN-Transformer network with efficient channel attention for organ segmentation in anatomical sectional images of canines.

Automated organ segmentation in anatomical sectional images of canines is crucial for clinical applications and the study of sectional anatomy. The manual delineation of organ boundaries by experts is a time-consuming and laborious task. However, semi-automatic segmentation methods have shown low segmentation accuracy. Deep learning-based CNN models lack the ability to establish long-range dependencies, leading to limited segmentation performance. Although Transformer-based models excel at establishing long-range dependencies, they face a limitation in capturing local detail information. To address these challenges, we propose a novel ECA-TFUnet model for organ segmentation in anatomical sectional images of canines. ECA-TFUnet model is a U-shaped CNN-Transformer network with Efficient Channel Attention, which fully combines the strengths of the Unet network and Transformer block. Specifically, The U-Net network is excellent at capturing detailed local information. The Transformer block is equipped in the first skip connection layer of the Unet network to effectively learn the global dependencies of different regions, which improves the representation ability of the model. Additionally, the Efficient Channel Attention Block is introduced to the Unet network to focus on more important channel information, further improving the robustness of the model. Furthermore, the mixed loss strategy is incorporated to alleviate the problem of class imbalance. Experimental results showed that the ECA-TFUnet model yielded 92.63% IoU, outperforming 11 state-of-the-art methods. To comprehensively evaluate the model performance, we also conducted experiments on a public dataset, which achieved 87.93% IoU, still superior to 11 state-of-the-art methods. Finally, we explored the use of a transfer learning strategy to provide good initialization parameters for the ECA-TFUnet model. We demonstrated that the ECA-TFUnet model exhibits superior segmentation performance on anatomical sectional images of canines, which has the potential for application in medical clinical diagnosis.

Phosphorylation of OsRbohB by the protein kinase OsDMI3 promotes H2O2 production to potentiate ABA responses in rice.

In rice, the Ca2+/calmodulin-dependent protein kinase OsDMI3 is an important positive regulator of abscisic acid (ABA) signaling. In ABA signaling, H2O2 is required for ABA-induced activation of OsDMI3, which in turn increase H2O2 production. However, how OsDMI3 regulates H2O2 production in ABA signaling remains unknown. Here we show that OsRbohB is the main NADPH oxidase involved in ABA-induced H2O2 production and ABA-mediated physiological responses. OsDMI3 directly interacts with and phosphorylates OsRbohB at Ser-191, which is OsDMI3-mediated site-specific phosphorylation in ABA signaling. Further analyses revealed that OsDMI3-mediated OsRbohB Ser-191 phosphorylation positively regulates the activity of NADPH oxidase and the production of H2O2 in ABA signaling, thereby enhancing the sensitivity of seed germination and root growth to ABA and plant tolerance to water stress and oxidative stress. Moreover, we discovered that the OsDMI3-mediated OsRbohB phosphorylation and H2O2 production is dependent on the sucrose non-fermenting 1-related protein kinases SAPK8/9/10, which phosphorylate OsRbohB at Ser-140 in ABA signaling. Taken together, these results not only reveal an important regulatory mechanism that directly activates Rboh for ABA-induced H2O2 production but also uncover the importance of this regulatory mechanism in ABA signaling.

Genome analysis of a plasmid-bearing myxobacterim Myxococcus sp. strain MxC21 with salt-tolerant property.

Myxobacteria are widely distributed in various habitats of soil and oceanic sediment. However, it is unclear whether soil-dwelling myxobacteria tolerate a saline environment. In this study, a salt-tolerant myxobacterium Myxococcus sp. strain MxC21 was isolated from forest soil with NaCl tolerance >2% concentration. Under 1% salt-contained condition, strain MxC21 could kill and consume bacteria prey and exhibited complex social behaviors such as S-motility, biofilm, and fruiting body formation but adopted an asocial living pattern with the presence of 1.5% NaCl. To investigate the genomic basis of stress tolerance, the complete genome of MxC21 was sequenced and analyzed. Strain MxC21 consists of a circular chromosome with a total length of 9.13 Mbp and a circular plasmid of 64.3 kb. Comparative genomic analysis revealed that the genomes of strain MxC21 and M. xanthus DK1622 share high genome synteny, while no endogenous plasmid was found in DK1622. Further analysis showed that approximately 21% of its coding genes from the genome of strain MxC21 are predominantly associated with signal transduction, transcriptional regulation, and protein folding involved in diverse niche adaptation such as salt tolerance, which enables social behavior such as gliding motility, sporulation, and predation. Meantime, a high number of genes are also found to be involved in defense against oxidative stress and production of antimicrobial compounds. All of these functional genes may be responsible for the potential salt-toleration. Otherwise, strain MxC21 is the second reported myxobacteria containing indigenous plasmid, while only a small proportion of genes was specific to the circular plasmid of strain MxC21, and most of them were annotated as hypothetical proteins, which may have a direct relationship with the habitat adaptation of strain MxC21 under saline environment. This study provides an inspiration of the adaptive evolution of salt-tolerant myxobacterium and facilitates a potential application in the improvement of saline soil in future.

A Retrospective Study on Risk Factors for Urinary Tract Infection in Patients with Intracranial Cerebral Hemorrhage.

OBJECTIVE: This study aimed to explore the risk factors of urinary tract infection (UTI) in patients with intracranial cerebral hemorrhage (ICH). DESIGN: This is a retrospective study, and a total of 77 patients with ICH consecutively admitted to the First Affiliated Hospital of USTC (Anhui Provincial Hospital, Hefei, China) during the period of August 2015 to August 2017 were included. The patients were divided into an UTI group (24 cases) and a non-UTI group (53 cases); patients with UTI were diagnosed according to clinical manifestations, recent urinary routines, and urine culture results. The following information in these two groups was recorded: age, sex, course of disease, side of paralysis, location and type of cerebral hemorrhage, disturbance of consciousness or not, the Brunnstrom stage of paralysed lower limbs, number of basic diseases, whether there were complications (tracheotomy, retention catheterization, pulmonary infection, pressure sore, deep venous thrombosis, etc.), whether rehabilitation interventions were conducted, blood routine, biochemistry index, DIC complete set, urine routine, and urine culture data. Univariate analysis and multivariate logistic regression analysis were used to examine the risk factors of UTI in patients with ICH. RESULTS: Univariate analysis showed that age, side of paralysis, disturbance of consciousness, the Brunnstrom stage of lower limbs, tracheotomies, retention catheterization, pulmonary infection, leukocyte count, neutrophil proportion, sodium, uric acid, D-dimer, and fibrinogen may be related to UTI in patients with ICH (P < 0.05). Regression analysis showed that age (OR (95% CI) = 1.207 (1.022-1.424), P < 0.05). Regression analysis showed that age (OR (95% CI) = 1.207 (1.022-1.424), P < 0.05). Regression analysis showed that age (OR (95% CI) = 1.207 (1.022-1.424), P < 0.05). Regression analysis showed that age (OR (95% CI) = 1.207 (1.022-1.424). CONCLUSIONS: Increased age and high D-dimer are independent risk factors for UTI in patients with ICH, while right-sided paralysis is a protective factor for UTI in patients with ICH.

Oxidative damage to DNA and its relationship with diabetic complications.

OBJECTIVE: To detect the oxidative DNA damage in diabetic patients and to investigate the relationship of oxidative DNA damage with diabetes and diabetic nephropathy. METHODS: Single cell gel electrophoresis (SCGE) was used to detect the DNA strand breaks in peripheral blood lymphocytes, and oxidative DNA damage product and serum 8-OHdG were determined by a competitive ELISA in 47 cases, including 25 patients without diabetic complications, 22 patients with diabetic nephropathy and 25 normal control subjects. RESULTS: Diabetic patients showed greater oxidative damage to DNA. The percentage of comet cells and the length of DNA migration (comet tail length) of peripheral blood lymphocytes were significantly increased in patients with diabetes, and significantly higher in patients with diabetic nephropathy than in diabetic patients without vascular complications (P < 0.05). There was a significant increase in serum 8-OHdG in diabetic patients compared with normal subjects (P < 0.05). Moreover, serum 8-OHdG was much higher in patients with diabetic nephropathy than in diabetic patients without vascular complications (P < 0.05). CONCLUSION: There is severe oxidative DNA damage in diabetic patients. Enhanced oxidative stress may be associated with diabetes, especially in patients with diabetic nephropathy.

[Investigation of RHD 1227A allele in five pedigrees in Zhejiang Han population].

OBJECTIVE: To analysis the genetic mode of Rh DEL phenotype and RHD 1227A allele in Zhejiang Han population through family investigations. METHODS: Rh DEL phenotypes were identified by a serologic adsorption-elution method. Two polymerase chain reaction-sequence specific prime (PCR-SSP) methods which detectED RHD 1227A allele and Rhesus hybrid box, respectively, and a nucleotide sequencing method focused on the exon 9 of RHD 1227A allele were employed to determine the zygosity of RHD allele. RESULTS: All five probands with Rh DEL phenotype harbored a RHD 1227A allele and had a RHD allele deletion, and they were RHD 1227A/RHd heterozygote. One of the parent members was found to contain a RHD 1227A allele and a normal RHD allele in pedigree 1, 2 and 3, respectively. Thus, they were RHD 1227A/RHD heterozygotes and presented normal D positive phenotype. The son of proband No 1. inherited the RHD 1227A allele and presented a normal D positive phenotype due to a RHD 1227A/RHD heterozygote; The offsprings of proband No. 2, No. 4, and No. 5 did not inherit RHD 1227A allele and presented a normal D positive phenotype. CONCLUSION: RHD 1227A allele is an important genetic marker of Rh DEL phenotype; RHD 1227A is recessive to normal RHD allele and dominant to RHd allele; RHD 1227A allele is an ancestral, but not a spontaneously mutated allele.

Metabolite Profiles of Icariin in Rat Feces, Bile and Urine by Ultraperformance Liquid-Chromatography/Quadrupole-Time-of-Flight Mass Spectrometry.

In this work, the metabolite profiles of icariin in rat feces, bile and urine were qualitatively investigated, and the possible metabolic pathways of icariin were subsequently proposed. After oral administration of icariin at a single dose of 100 mg/kg, rat biological samples were collected and pretreated. Then, these pretreated samples were detected by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). In all, 17 metabolites were identified in the biosamples. Of these, 5, including F8-F9 (icariside I), D3-D4 (isopentenyl alcohol-icaritin-3-O-rha-7-O-gluA) and N3 (1,3-isoprene icariside II), were to our knowledge reported for the first time. The results indicated that icariin was metabolized via desugarization, dehydrogenation, hydroxylation, demethylation and glucuronidation pathways in vivo. This study revealed the possible metabolite profiles of icariin in rats.