Effects of S1P/S1PR1 pathway on high glucose induced epithelial-mesenchymal transition of rat renal tubular epithelial cells by regulating ROS/NLRP3 / 中国药理学通报

Aim To explore the effect of S1P/S1PR1 signaling pathway on high glucose(HG)-induced epithelial-mesenchymal transition of rat renal tubular epithelial cells and its possible mechanism. Methods Cells were treated with different concentrations of glucose, and intracellular S1P expression was detected by ELISA and S1PR1 protein expression was detected by Western blot. The cells were divided into normal control group, HG group and HG + siS1PR1 group. The expression of E-cadherin, Vimentin, Fibronectin and Twist mRNA were detected by RT-qPCR and E-cadherin, α-SMA, Vimentin, NLRP3, ASC and NF-κB protein expression were detected by Western blot, and the levels of reactive oxygen species(ROS) were detected by flow cytometry. The cells were divided into normal control group, S1P group and S1P + siS1PR1 group. Vimentin, Snail, α-SMA, NLRP3, ASC and NF-κB protein expressions were detected by Western blot, and ROS levels were measured by fluorescence microscopy. Results ELISA results showed that the content of S1P in cells increased significantly under high glucose stimulation. Western blot results showed that S1PR1 protein expression was significantly higher at 30 mmol · L

Advances in online two-dimensional liquid chromatography for natural product screening / 药学学报

As a treasure resource of novel drug lead compounds, how to rapidly and high-efficiently screen and isolate active components from natural products is critical. Thanks to its high resolution, high automation and flexible integration, online two-dimensional liquid chromatography has great potential for screening active ingredients from complex matrices by integrating a highly specific bio-recognition process into a two-dimensional liquid chromatography system before, on or after the column separation. This review comprehensively summarized recent developments, applications and shortcomings of online two-dimensional liquid chromatography for natural product screening from different integration modes, including pre-column, on-column and post-column screening methods.

An Anterior Cingulate Cortex-to-Midbrain Projection Controls Chronic Itch in Mice / 神经科学通报·英文版

Itch is an unpleasant sensation that provokes the desire to scratch. While acute itch serves as a protective system to warn the body of external irritating agents, chronic itch is a debilitating but poorly-treated clinical disease leading to repetitive scratching and skin lesions. However, the neural mechanisms underlying the pathophysiology of chronic itch remain mysterious. Here, we identified a cell type-dependent role of the anterior cingulate cortex (ACC) in controlling chronic itch-related excessive scratching behaviors in mice. Moreover, we delineated a neural circuit originating from excitatory neurons of the ACC to the ventral tegmental area (VTA) that was critically involved in chronic itch. Furthermore, we demonstrate that the ACC→VTA circuit also selectively modulated histaminergic acute itch. Finally, the ACC neurons were shown to predominantly innervate the non-dopaminergic neurons of the VTA. Taken together, our findings uncover a cortex-midbrain circuit for chronic itch-evoked scratching behaviors and shed novel insights on therapeutic intervention.

Study on Platelet Adhesion and Aggregation Induced by Gradient Shear Stress Using Microfluidic Chip Technology / 中国实验血液学杂志

OBJECTIVE@#To study the effect of gradient shear stress on platelet aggregation by microfluidic chip Technology.@*METHODS@#Microfluidic chip was used to simulate 80% fixed stenotic microchannel, and the hydrodynamic behavior of the stenotic microchannel model was analyzed by the finite element analysis module of sollidwork software. Microfluidic chip was used to analyze the adhesion and aggregation behavior of platelets in patients with different diseases, and flow cytometry was used to detect expression of the platelet activation marker CD62p. Aspirin, Tirofiban and protocatechuic acid were used to treat the blood, and the adhesion and aggregation of platelets were observed by fluorescence microscope.@*RESULTS@#The gradient fluid shear rate produced by the stenosis model of microfluidic chip could induce platelet aggregation, and the degree of platelet adhesion and aggregation increased with the increase of shear rate within a certain range of shear rate. The effect of platelet aggregation in patients with arterial thrombotic diseases were significantly higher than normal group (P<0.05), and the effect of platelet aggregation in patients with myelodysplastic disease was lower than normal group (P<0.05).@*CONCLUSION@#The microfluidic chip analysis technology can accurately analyze and evaluate the platelet adhesion and aggregation effects of various thrombotic diseases unde the environment of the shear rate, and is helpful for auxiliary diagnosis of clinical thrombotic diseases.

CRISPR/Cas9 Targeted Editing of Osiris24 in Drosophlia melanogaster / 中国生物化学与分子生物学报

The gene expression of Osiris is coincident with the timing of chitin deposition. Osiris gene may be involved in the developmental regulation of insect cuticle. The objective of this study is to generate the gene-edited flies with Osiris24 by CRISPR/Cas9-mediated editing system to understand the traits of Osiris24 mutant flies and the expression pattern of Osiris24. Two sgRNA targeted sequences were designed according to the sequence of exon 1 of Osiris24 and inserted into pCFD4 vector backbone. A donor vector with Gal4 protein sequence was constructed. Above two plasmids were mixed and injected into nosCas9 fly embryos to generate GO generation. The results showed that 92.8% GO flies have Gal4 protein insert in genome. Homozygous mutants of Osiris24 were lethal at the embryonic stage or first-instar stage, and no visible phenotype was observed in heterozygous mutants. Osiris24 is expressed throughout larval and pupal stages. At the larval stage, Osiris24 is mainly expressed in the integument, foregut and hind-gut, while Osiris24 is expressed in the integument and wings at the pupal stage. These results indicated that Osiris24 plays an important role in the development of Drosophila. This study provides a research model for in-depth exploration of Osiris gene function.

Intermedin inhibits lipopolysaccharide-induced polarization of macrophages by activating AMPK signaling pathway / 中国药理学通报

Aim To evaluate the mechanism by which intermdin(IMD)inhibits lipopolysaccha ride(LPS)-induced polarization in RAW264.7 cells.Methods RAW264.7 cells were divided into control groups, LPS groups, LPS+IMD groups, LPS+IMD+Compound C groups.The mRNA expressions of tumor necrosis factor-α,(TNF-α,), CD86, inducible nitric oxide synthase(iNOS), Arginase-1(Arg-1)and CD206 were detected by Realtime-PCR.The protein expressions of p-AMPK, AMPK, TNF-α, intereukin-6(IL-6)and intereukin-10(IL-10)were detected by Western blot.The proportion of CD86+ M1 type cells was detected by Flow cytometry.In addition, the expression levels of supernatant cytokines, including IL-6 and TNF-α, were detected by ELISA.Results Compared with control and LPS groups, IMD treatment could up-regulate the expression level of p-AMPK and the ratio of p-AMPK/AMPK.LPS promoted M1 polarization, since the expressions of CD86, TNF-α and iNOS increased, while the expressions of CD206 and Arg-1 decreased by LPS induction.The proportion of M1 type cells increased and the secretion of TNF-α, IL-6 in the cell supernatant increased.And IMD treatment could inhibit the polarization of M1 induced by LPS.These effects were reversed by Compound C, an inhibitor of AMPK.Conclusion IMD can inhibit the M1-type polarization of LPS-induced macrophages by activating AMPK signaling pathway.

Study on improvement of non-alcoholic steatohepatitis by regulating CDKN1A by Yiqi-Bushen-Tiaozhi formula / 中国药理学通报

Aim To screen the mechanism of Yiqi- Bushen-Tiaozhi formula ( YBTF) in treating non-alcoholic steatohepatitis ( NASH) by network pharmacology analysis and to verify it by animal experiments. Methods TCMSP database and HPLC-MS analysis were used to mine the active ingredients and targets of YBTF; GSE89632 dataset was used to screen the differential expressed genes ( DEGs) between the normal and the NASH groups; GeneCards and DisGeNET databases were used to screen NASH-related disease genes. The intersection genes of the three are the target genes of YBTF treatment of NASH. The intersection gene of the three sets of genes was the target gene of YBTF in treating NASH. GO, KEGG, DO enrichment analysis, protein-protein interaction network, and network topology analysis were used to identify the hub genes of YBTF in the treatment of NASH. Molecular docking was used to judge whether cmcial target genes, active ingredients could be combined and exer ted a curative effect; Oil red 0 and HE staining were used to determine whether YBTF could treat NASH mice; (3-galactosidase ( SA- (3-Gal) test was used to determine whether NASH mice had hepatocyte senescence and whether YBTF improved senescence; West-ern blot. Quantitative Real-time PGR ( qRT-PCR) combined with sequencing results were used to verify whether YBTF could regulate the expression of the essential target genes screened from the protein and RNA levels. Results YBTF could improve cellular aging and treat NASH through CDKN1A. Conclusion The rational application of Traditional Chinese medicine (TCM) network pharmacology and experiments can provide new ideas and directions for studying the mechanism of YBTF.

Advances in understanding leukemia bone marrow microenvironments / 药学学报

The bone marrow microenvironment, also known as the bone marrow niche, plays a critical role in maintaining the functions of hematopoietic stem cells. Under physiological conditions, various bone marrow cells regulate each other to sustain hematopoietic homeostasis. However, bone marrow cells gain abnormal function under pathological conditions to cause and promote the occurrence of leukemia and induce drug resistance. Recent findings indicate that abnormal proliferation and differentiation are not the sole reason to cause leukemia. Different types of bone marrow cells also induce intercellular adhesion, abnormally secrete cytokines and chemokines, accelerating leukemia's progress. This article reviews the multiple signaling pathways that regulate the formation and progress of leukemia bone marrow niche, such as C-X-C motif chemokine ligand 12/C-X-C motif chemokine receptor 4 signaling pathway, et al. It emphasizes that targeting leukemia bone marrow niche is a vital strategy for improving the leukemia treatment.

Acetylation of Rehmannia glutinosa polysaccharides and antioxidant activity of acetylated derivatives / 中国中药杂志

This study aims to acetylate Rehmannia glutinosa polysaccharides by acetic anhydride method, optimize process parameters and evaluate their antioxidant activity. With the degree of substitution(D_s) as a criterion, the effects of reaction time, acetic anhydride-to-polysaccharides ratio and temperature were investigated. Process parameters were optimized by single-factor experiment and response surface methodology. The infrared spectroscopy(IR) and scanning electron microscopy(SEM) proved the successful acetylation and were employed to preliminarily analyze the structural characteristics of acetylated derivatives. The results showed that the D_s was 0.327 under the optimal technological conditions, including m(acetic anhydride):m(R. glutinosa polysaccharides)=2.70, reaction time 3.0 h and temperature 48 ℃. Further, the antioxidant properties of acetylated derivatives were investigated in vitro and acetylation was found effective to improve the antioxidant activity of R. glutinosa polysaccharides. This study provides a reference for the further development and application of R. glutinosa polysaccharides.

Structure Analysis of the Interfering Substance N-methyl-2-phenylpropan-1-amine of Methamphetamine in Wastewater / 法医学杂志

OBJECTIVES@#To analyze the chemical structure of the interfering substance that affects the result of methamphetamine analysis in wastewater.@*METHODS@#A combination of GC-MS and liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) was used to analyze the mass spectrum characteristics of the interfering substance that affects the result of methamphetamine analysis and to infer its possible structure. Liquid chromatography-triple quadrupole-mass spectrometry (LC-TQ-MS) was used to confirm the control material.@*RESULTS@#Using LC-QTOF-MS in positive electrospray ionization (ESI+) mode, the mass-to-charge ratio (m/z) of quasi-molecular ion in the MS1 mass spectrometry of interfering substance was identical to that of methamphetamine, indicating that the interfering substance was probably an isomer of methamphetamine. The MS2 mass spectra obtained at three collision energies of 15 V, 30 V and 45 V were highly similar to methamphetamine, suggesting that the interfering substance contained methylamino and benzyl groups. Further analysis using GC-MS in electron impact (EI) ionization mode showed that the base peak in the mass spectrum of the interfering substance was at m/z 44. The interfering substance was confirmed to be N-methyl-2-phenylpropan-1-amine by compared with the standard reference.@*CONCLUSIONS@#The chemical structure of N-methyl-2-phenylpropan-1-amine is highly similar to methamphetamine, which is easy to cause interference for the detection of trace amounts of methamphetamine in wastewater using LC-TQ-MS. Therefore, in the actual analysis, the chromatographic retention time can be used to distinguish between N-methyl-2-phenylpropan-1-amine and methamphetamine.

Establishment of high-resolution bioassay profiling platform to screen α-glucosidase inhibitors from Malus hupehensis / 药学学报

italic>α-Glucosidase inhibitors play an important role in the treatment of diabetes. This study established a high-resolution bioassay profiling platform for rapidly screening α-glucosidase inhibitors in natural product extracts. Five α-glucosidase inhibitors were identified from Malus hupehensis, namely, 3-hydroxyphloridzin, quercetin-3-O-β-D-glucopyranoside, phloridzin, avicularin and quercitrin. The establishment and successful application of this platform provides a powerful tool for the efficient discovery of anti-diabetic active ingredients in complex systems.

New tirucallane-type triterpenoids from the resin of Boswellia carteriiand their NO inhibitory activities / 中国天然药物

Six new tirucallane-type triterpenoids (1-6), along with ten known triterpenoids, were isolated from methylene chloride extract of the resin of Boswellia carterii Birdw. By the application of the comprehensive spectroscopic data, the structures of the compounds were clarified. The experimental electronic circular dichroism spectra were compared with those calculated, which allowed to assign the absolute configurations. Compounds 5 and 6 possesed a 2, 3-seco tirucallane-type triterpenoid skeleton, which were first reported. Their inhibitory activity against NO formation in LPS-activated BV-2 cells were evaluated. Compound 9 showed appreciable inhibitory effect, with an IC

Mechanism of Hispolon in treatment of colon cancer based on network pharmacology and cell experiment / 中国药理学通报

Aim To explore the mechanism of Hispolon in the treatment of colon cancer by network pharmacology and cell experimental validation. Methods The potential targets of Hispolon were obtained from the Swiss Target Prediction website, and intersected with colon cancer targets from GeneCards and OMIM databases. The protein-protein interaction network of targets was built by the STRING11. 0 database. Meanwhile , the core targets of PPI network was explored by Cytoscape 3. 7. 2 software. Furthermore, the GO and KEGG pathway enrichment were analyzed by Metas- cape database. Finally, Western blotting was used to verify the regulation of Hispolon on some key targets in colon cancer cell SW480. Results Sixty-nine com-mon targets of Hispolon and colon cancer were obtained, which were colon cancer therapeutic targets. The core targets included BCL-2L1, EP300, CDK1, AR, MTOR and EGFR. The enrichment analysis showed that Hispolon played a role in the treatment of colon cancer by regulating the pathways in cancer, PI3K-Akt signaling pathway, prostate cancer and Mi- croRNAs in cancer. And the key targets in the pathway involved core targets such as BCL-2 LI, EP300, CDK1, MTOR and EGFR. Cell experiments confirmed that Hispolon promoted SW480 cell apoptosis by down- regulating the expression of target proteins BCL-2L1 and mTOR. Conclusions The discussion of the molecular mechanism of Hispolon in the treatment of colon cancer suggests that Hispolon may play a role in the treatment of colon cancer through multiple targets and multiple pathways. The results provide a scientific basis for the elucidation of the mechanisms and clinical application of Hispolon against colon cancer.

Study on chemical constituents from fruiting bodies of Ganoderma calidophilum / 中国中药杂志

Chemical constituents were isolated and purified from fruiting bodies of Ganoderma calidophilum by various column chromatographic techniques, and their chemical structures were identified through combined analysis of physicochemical properties and spectral data. As a result, 11 compounds were isolated and identified as(24E)-lanosta-8,24-dien-3,11-dione-26-al(1), ganoderone A(2), 3-oxo-15α-acetoxy-lanosta-7,9(11), 24-trien-26-oleic acid(3),(23E)-27-nor-lanosta-8,23-diene-3,7,25-trione(4), ganodecanone B(5), ganoderic aldehyde A(6), 11β-hydroxy-lucidadiol(7), 3,4-dihydroxyacetophenone(8), methyl gentiate(9), ganoleucin C(10), ganotheaecolumol H(11). Among them, compound 1 is a new triterpenoid. The cytotoxic activities of all of the compounds against tumor cell lines were evaluated. The results showed that compounds 1, 3, 4 and 6 showed cytotoxic activity against BEL-7402, with IC_(50) values of 26.55, 11.35, 23.23, 18.66 μmol·L~(-1); compounds 1 and 3-6 showed cytotoxic activity against K562, with IC_(50) values of 5.79, 22.16, 12.16, 35.32, and 5.59 μmol·L~(-1), and compound 4 showed cytotoxic activity against A549, with IC_(50) value of 42.50 μmol·L~(-1).

Herbal textual research of Astragali Complanati Semen / 中草药

Astragali Complanati Semen has a long-term history of use as a common Chinese traditional and herbal medicine. Through a herbal textual research on the appellation, origin, efficacy, genuine producing area and counterfeit of Astragali Complanati Semen, the evolution of its appellation underwent three stages of "white terrestris", "Shayuan terrestris", and "Astragali Complanati Semen"; The botanical origin and medicinal part is dried mature seed from Astragalus complanatus of Legume family recorded in the literatures of past dynasties; And the varieties of ancient and modern application are basically the same. Its kidney-nourishing and essence-enriching efficacy has been progressively cleared with clinical application since records began in Bencao Yanyi in the Song dynasty; The genuine producing areas of Astragali Complanati Semen were Shayuan in Shaanxi Province from the Song Dynasty, and gradually moved to Tongguan in Shaanxi Province and Shanxi Province in the Ming and Qing Dynasties. After the founding of new China, it was mainly distributed in Shaanxi Province and then spread to northern adjacent provinces, in which the Tongguan in Shaanxi Province was preferred. Its counterfeits included Mapiao semon, an unknown counterfeit with a hint of green and Astragalus sinicus seed based on herbal literatures published before 1949, and mainly included the seeds of A. sinicus, Crotalaria pallida, Asfraglus chinensis and Astragalus adsurgens after 1949. Based on a systematic herbal textual research, this article takes a radical reform of Astragali Complanati Semen, so as to provide reference for its further development and utilization.

Advances in chromatography-based methods for screening active compounds from natural products / 药学学报

Natural products have been a major source of leading compounds in drug discovery. How to effectively screen active compounds from complex matrix remains an interesting topic. In this review, we comprehensively summarized advanced liquid chromatography based approaches in natural products screening, including pre-column, on-column and post-column screening methods. Their advantages, disadvantages and prospect are also discussed.

Clinical Phenotype and Gene Mutation Analysis of a Family with Early-Onset Familial Alzheimer′s Disease / 中山大学学报(医学科学版)

@#【Objective】A full exome sequencing of an early-onset family Alzheimer′s disease (EOFAD) was conduct? ed to identify the mutational sites which may cause diseases. The result of the current study may provide suggestion to genetic counseling and prenatal diagnosis.【Methods】Whole exome sequencing was performed on the family members and software PolyPhen-2 as well as SIFT was employed for hazard prediction (Prediction on functional effects of the missense mutation).【Results】The heterozygous mutation c.758A>G (p.Tyr253Cys) in exon 9 of TTC3 gene had been identified in proband whose mother had been proved with heterozygous mutation c.758A>G. According to the family separation and related bioinformatics analysis, the mutant gene was a possible pathogenic mutation. 【Conclusion】 A new mutation was found of c.758A>G in TTC3 gene within a Chinese EOFAD family and a new mutation to the spectrum of genetic mutation in EOFAD was expanded. The finding provides a significant groundwork for future exploration on the mechanisms underlying EOFAD.

Research advances in the functional roles of ion channels in immune cells and immune response / 生理学报

Ion channels are a widespread class of membrane proteins that help establish and control cell membrane potential by allowing the passive diffusion of inorganic ions with high specificity through cell membrane. They are widely distributed in various cells and tissues, and their normal structure and function are of fundamental importance for all living organisms. The rapid advances in molecular cloning, protein structure analysis, patch clamp recordings and other technologies have greatly promoted the research on the biophysical and molecular properties of ion channels, and made significant progress in the study of the relationship between ion channels and pathophysiology as well. The immune system is made up of immune cells and organs that work together to protect the body and respond to infection and disease. Remarkably, recent basic and clinical research has revealed that ion channels are frequently and abundantly expressed in immune cells and have crucial roles in immune cell development and immune response. This review summarized recent progress in the roles of ion channels in immune cells, including the expression and regulation of ion channels in immune cells, the effects of ion flux mediated by ion channels on lymphocyte development, and functional roles of ion channels in both innate and adaptive immune responses. We also discussed some unresolved and insufficiently addressed issues in the current research, so as to provide an informative reference for better understanding the functional roles of ion channels in the immune system and further elucidation of their function from a physiological and pathological point of view.

Diagonal root separation method for extracting impacted double-root mandibular third molars / 华西口腔医学杂志

Double-root mandibular third molars are the common types of mandibular third molars that are prone to impac-tion. Based on the summary of clinical cases and experience, the commonly used crown-root separation technique is improved as diagonal root separation technique that divides the roots by the line connecting the distal axis angle to the root furcation. This new method improves the efficiency of the operation and reduces surgical trauma. This paper details the surgical method, minimally invasive mechanics, and key points of this technique, compares the technique with the crown-root separation tech-nique, and discusses their joint applications.

Association between obesity and blood pressure in preschool children in urban areas / 中华疾病控制杂志

Objective To investigate the blood pressure status of preschool children in urban areas of Qingdao, and to determine the relationship between obesity and blood pressure in preschool children. Methods A stratified cluster sampling method was used to select a total of 13 kindergartens in urban districts of Qingdao. Height, weight, waist circumference, hip circumference and blood pressure of children in three classes were measured. Body mass index (BMI), waist to hip ratio and waist to height ratio were calculated and the relationship between obesity and blood pressure was analyzed. Results The mean values of systolic and diastolic blood pressure in preschool children in urban areas of Qingdao were (95.52±7.66) and (62.78±6.52) mmHg, respectively.The detection rate of hypertension in preschool children was 13.50%. The SBP and DBP were positively correlated with BMI, waist circumference, hip circumference and waist to height ratio. There was a linear regression relationship between body mass index and age and blood pressure. The risk of hypertension in overweight and obese children was 5.191 and 2.824 times of normal body weight, respectively. Conclusions The prevalence of hypertension in preschool children in Qingdao urban areas is high.Overweight and obesity are risk factors for elevated blood pressure.Therefore, while preventing preschool children from obesity, preschool children's blood pressure monitoring and blood pressure monitoring and early intervention of hypertension of preschool children should be implemented.