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.

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.

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.

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.

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

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).

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.

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.

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.

Inhibition of compound kushen injection on suppressing proliferation, migration and invasion of SMMC-7221 cells in vitro and its mechanism / 中国药理学与毒理学杂志

OBJECTIVE To explore the inhibition of Compound Kushen Injection (CKI) on the prolif-eration, in vitro migration and in vitro invasion of SMMC-7221 cells and the mechanism. METHODS SMMC-7221 cells were treated with CKI of different dilution percentages (0.0% (cell control), 2.5%, 5.0% and 10.0%) for 24 h. The viability of SMMC-7221 cells was evaluated by MTT assay, while the ability of cell migration and invasion was evaluated by the wound healing and transwell chamber assay, respectively. Cells were treated with 0.0%, 5.0% and 10.0% CKI for 12 or 24 h, before the mRNA and protein levels of NF-kB were detected by RT-PCR and Western blotting, respectively. RESULTS Compared with the cell control group, the survival rates in 2.5%, 5.0% and 10.0% CKI groups were reduced to (92±7)%, (77±5)% (P<0.01) and (56±7)% (P<0.01), the migration rates to (90±7)%, (50±10)% (P<0.01) and (25±5)% (P<0.01), and the invasion rates to (98±7)%, (51 ±10)% (P<0.01) and (20±5)% (P<0.01), respectively. The NF-kB mRNA levels in 5.0% and 10.0% CKI groups were reduced to (42±9)% and (46±10)% of the cell control group (P<0.01) after 12 h treatment of CKI, and the protein levels of NF-kB were reduced to (67±16)% and (27±11)% of cell control group after 24 h treatment with CKI, respectively (P<0.01). CONCLUSION CKI can inhibit the proliferation, migration and invasion of SMMC-7221 cells in vitro, and the mechanism may be related to the inhibition of NF-kB signal pathway.

Protective effect of cardiomyopeptide on doxorubicin induced cell injury in H9c2 cardiomyocytes / 中国药理学通报

Aim To investigate the effect of cardio-myopeptide on doxorubicin-induced toxicity on H9c2 cardiomyoblasts and its related mechanism. Methods H9c2 cells were respectively pretreated with different concentrations(0, 10, 20, 40 mg ∗ L-1) of cardio-myopeptide for 6 h,8 h, 12 h. Cell viability was determined by MTT assay. The protein expression of caspase-3, Bcl-2, Bax, IGF-1R and IGFBP-3 were detected by Western blot. Results Cardiomyopeptide protected H9c2 cardiomyocytes from doxorubicin induced apoptosis in a dose-and time-dependent manner. The half maximal inhibitory concentration (IC50) was shifted from(1.2 ±0.4) umol • L-1 to(2.3 ±0.2) jimol • L-1 The expression of Bcl-2, IGF-1R was up-regulated , and Bax, IGFBP-3 and caspase-3 decreased in H9c2 cells. Conclusions Cardiomyopeptide could prevent from apoptosis induced by doxorubicin in H9c2 cells via increasing expression of IGF-1R, thus up-reg-ulation of the antiapoptotic protein Bcl-2 and inhibiting caspase-3 activity.

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.

Effect of different interfaces on FiO_2 and CO_2 rebreathing during noninvasive positive pressure ventilation / 中国实用内科杂志

OBJECTIVE: To investigate the effect of different interfaces on FiO_2 and CO_2 rebreathing(ViCO_2) during noninvasive positive pressure ventilation. METHODS: The Active Servo Lung respiratory simulation system was used to simulate a hypercapnia COPD patient received NPPV through injection of CO_2 gas from the place closest to the outlet of simulated lung to maintain end-tidal carbon dioxide partial pressure at 80 mmHg,and oxygen delivery during NPPV was simulated by means of injection of 5 L/min oxygen through the hole of interface. FiO_2 and ViCO_2 were integral calculated through the synchronous collect real-time pressure, flow rate, oxygen concentration and CO_2 concentration. The effect of different type and models of interfaces on FiO_2 and ViCO_2 were compared, and the bivariate correlation analysis was applied to investigate the correlation of FiO_2 and ViCO_2. RESULTS: The inner volume, leak volume and leak position of different interfaces were different. The FiO_2 of the oral-nasal mask and nasal mask were significantly different(P<0.05), which were(39.81±9.06)% and(43.91±6.33)%,respectively. The FiO_2 of the different models of interfaces was significantly different(P<0.05), the FiO_2 of the No.4 oral-nasal mask was the lowest(28.29±0.08)%, and the FiO_2 of the No. 10 oral-nasal mask was the highest(53.83±0.63)%. The ViCO_2 of the oral-nasal mask and nasal mask wasn't significantly different(P=0.19), which was(15.26±1.27)mL and(14.79±2.21)mL, respectively. The ViCO_2 of the different models of interfaces was significantly different(P<0.05), the FiO_2 of the No.8 nasal mask was the lowest(12.48±0.29%), and the ViCO_2 of the No. 10 nasal mask was the highest(18.38±0.31)%. There was a significant correlation between FiO_2 and ViCO_2(r=0.41, P<0.05), and the general linear equation was Y=14.51+1.82 X(R~2=0.168). CONCLUSION: Different interfaces have significant effects on FiO_2 and ViCO_2 during NPPV,and FiO_2 and ViCO_2 are positively correlated.

Study on DNA molecular identification of mix samples of five species of Baitouweng medicinal materials based on high-throughput sequencing technology / 药学学报

Traditional Chinese medicine Baitouweng have a long history of application. The pharmacopoeia included dry roots of Pulsatilla chinensis (Bge.) Regel of Ranunculaceae. There are easily confused species in the market circulation, such as P. cernua (Thunb.) Bercht. et Opiz., P. dahurica (Fisch.) Spreng., P. turczaninovii Kryl. et Serg., and P. chinensis (Bge.) Regel var. kissii (Mandl) S. H. Li et Y. H. Huang, etc. In this study, using the method of metagenomics, based on high-throughput sequencing technology, the ITS2 sequence of mixed samples of five species of Baitouweng medicinal materials was sequenced. First, the total DNA extraction of medicinal materials mixing powder, and the ITS2 fragment of total DNA was amplified by PCR. Second, the Illumina MiSeq platform was used to carry out Paired-end sequencing for DNA fragments. Last, using FLASH, QⅡME and GraPhlAn software to arrange and analyze, and clustering analysis with the sequences of uploaded to GenBank by our group in the early stage. The results showed that a total of 53 024 sequences of ITS2 were obtained from the mixed samples, there are 52 295 effective sequences, there are a total of 49 079 of five species of medicinal materials of P. Miller. After the representative sequences and the sequence of uploaded to GenBank by our group in the early stage were clustering analysis, 5 species of Baitouweng medicinal materials were clustered into one branch separately, presenting monophyletic. The results showed that using the high-throughput sequencing technology, using ITS2 sequence as DNA barcode, the mix powder of 5 species of Baitouweng medicinal materials could be effectively identified. It provides a new method and thought for the origin identification of mixed Chinese medicinal materials.

Studies on the enzymatic synthesis of a new PPT-type ginsenoside via UDP-glycosyltransferase PgUGT74AE2 from Panax ginseng / 药学学报

UDP-glycosyltransferase PgUGT74AE2 from Panax ginseng can transfer a glucose moiety to the free C-3 hydroxyl of protopanaxadiol (PPD) to produce ginsenoside Rh2. However, no report demonstrates that PgUGT74AE2 can transfer a glucose moiety to the free C-3 hydroxyl of protopanaxatriol (PPT) to produce a PPT-type ginsenoside. In this study, the expression plasmid pET-32a-PgUGT74AE2 was constructed for expression of the recombinant protein and transferred into Escherichia coli Transetta (DE3) to generate the recombinant strain Transetta-PgUGT74AE2. The recombinant enzyme PgUGT74AE2 was expressed by induction of isopropyl-β-D-thiogalactoside (IPTG). An in vitro enzymatic reaction system was established with the recombinant enzyme PgUGT74AE2 and the substrate PPT. PgUGT74AE2 catalyzed the glycosylation of the free C-3 hydroxyl of PPT to produce 3-O-β-D-glucopyranosyl-dammar-24-ene-3β,6α,12β,20S-tetraol, a new PPT-type ginsenoside. This study provides an efficient approach for the biosynthesis of a new PPT-type ginsenoside through in vitro enzymatic reaction, which may pave a way to produce promising lead in drug discovery.