Identification of the metabolites from co-cultures of marine Streptomyces sp. IMB18-531 and Cladosporium sp. IMB19-099 / 药学学报

A new siderophore chelate (1) and 8 known compounds were identified from the liquid co-cultures of the marine-derived Streptomyces sp. IMB18-531 and Cladosporium sp. IMB19-099 by a combination of chromatography methods, including C18 reversed-phase medium pressure chromatography, gel column chromatography and HPLC. Their structures were determined by spectroscopic analysis and chemical methods as aluminioxamine E (1), desferrioxamine E (2), ferrioxamine E (3), terragine E (4), capsimicin (5), cyclo(L-prolinyl-L-tyrosine) (6), anthranilic acid (7), (Z)-14-methylpentadec-9-enoic acid (8), and (Z)-hexadec-8-enoic acid (9). Compound 2 showed inhibitory activities against the expression of liver fibrosis related genes COL1A1, MMP2, and TIMP2. Compounds 5, 8, and 9 displayed antibacterial activities against methicillin-resistant Staphylococcus aureus, S. epidermidis and Bacillus subtilis, with MICs of 16-64 μg·mL-1. Compound 5 showed cytotoxicities against human pancreatic cancer MIA Paca-2 and human colon cancer HT-29 cell lines with IC50 of 2.9 and 6.3 μmol·L-1, respectively.

Influences of ethanol and temperature on sucrose-evoked response of gustatory neurons in the hamster solitary nucleus

Taste-responsive neurons in the nucleus of the solitary tract (NST), the first gustatory nucleus, often respond to thermal or mechanical stimulation. Alcohol, not a typical taste modality, is a rewarding stimulus. In this study, we aimed to investigate the effects of ethanol (EtOH) and/or temperature as stimuli to the tongue on the activity of taste-responsive neurons in hamster NST. In the first set of experiments, we recorded the activity of 113 gustatory NST neurons in urethane-anesthetized hamsters and evaluated responses to four basic taste stimuli, 25% EtOH, and 40°C and 4°C distilled water (dH2O). Sixty cells responded to 25% EtOH, with most of them also being sucrose sensitive. The response to 25% EtOH was significantly correlated with the sucrose-evoked response. A significant correlation was also observed between sucrose- and 40°C dH2O- and between 25% EtOH- and 40°C dH2O-evoked firings. In a subset of the cells, we evaluated neuronal activities in response to a series of EtOH concentrations, alone and in combination with 32 mM sucrose (EtOH/Suc) at room temperature (RT, 22°C–23°C), 40°C, and 4°C. Neuronal responses to EtOH at RT and 40°C increased as the concentrations increased. The firing rates to EtOH/Suc were greater than those to EtOH or sucrose alone. The responses were enhanced when solutions were applied at 40°C but diminished at 4°C. In summary, EtOH activates most sucrose-responsive NST gustatory cells, and the concomitant presence of sucrose or warm temperatures enhance this response. Our findings may contribute to elucidate the neural mechanisms underlying appetitive alcohol consumption.

SNP of AHSA2 gene in three cattle breeds using snapshot technology

Droughtmaster is a tropical breed of beef cattle that can survive in hot climates and easily adapt to torrid environments. These traits are important in livestock breeding. In this study, we genotyped five single-nucleotide polymorphisms (SNPs) of the AHSA2 gene from 190 cattle belonging to three different breeds (Droughtmaster, Angus and Simmental) by using snapshot technology. This work aimed to identify the valuable molecular marker of heat resistance in cattle. Results showed that Droughtmaster exhibited higher expected heterozygosity and polymorphic information content compared with the two other breeds. The AHSA2-1 locus deviated from the Hardy– Weinberg equilibrium in the Droughtmaster breed (P \ 0.05). Two SNPs in Droughtmaster diverged significantly from Angus and Simmental. The SNPs were identified as AHSA2-3 and AHSA2-4, which were closely linked to the three breeds based on pair-wise FST. AHSA2-4 involved a missense mutation. In summary, the GG genotypes in AHSA2-3 and AHSA2-4 may be candidate genotypes associated with heat resistance traits and may serve as valuable genetic markers for breeding of heat-tolerant beef cattle in the future.

Effect of Total Alkaloids from Nandina domfestica in Attenuating Arsenic Trioxide and Its LC-MS / 中国实验方剂学杂志

Objective::To study the effect of total alkaloids of Nandina domestica in attenuating arsenic trioxide, and analyze chemical constituents of alkaloids extract of Nandina, in order to provide the theoretical basis for studying the effect of N. domestica in attenuating arsenic trioxide with alkaloids extract of N. domestica as effective fraction. Method::The model of acute liver injury induced by arsenic trioxide was used to compare the effects on heart and liver functions of mice between arsenic trioxide alone and total alkaloids of N. domestica combined with arsenic trioxide. The detoxification of total alkaloids on arsenic trioxide was evaluated based on biochemical parameters and pathological report. Peakview (Version1.2, AB SCIEX) software was used to process the mass spectrometry data of total alkaloids of N. domestica. The structures of determined compounds were identified by molecular weight of compound (molecular formula), secondary fragments, chromatographic peak retention and literature information. Result::Among biochemical indicators, creatine kinase(CK), lactate dehydrogenase(LDH), blood urea nitrogen(BUN), malondialdehyde(MDA), alanine aminotransferase(ALT) and amino transferase of aspartate(AST) of the arsenic trioxide group were increased, while elimination rates of Na+ -K+ -adenosine triphosphate(ATP), superoxide dismutase(SOD), catalase(CAT) and serum creatinine(SCr) were decreased, compared with those of the combination group. CK and LDH of the alkaloids extract group were more obviously increased than those of the N. domestica extract group, but with no remarkable difference. In histomorphometric examination, edema of mouse heart cells was improved, and some kidney and liver damages in rats were alleviated. Totally 25 alkaloids of alkaloid extract were identified. Among them, 18 were known, and 7 were unknown, including 3 structural types, in which apomorphine alkaloids were mostly. Conclusion::Heart, kidney and liver damage degrees of the combination group were significantly alleviated compared with the arsenic trioxide group. The total alkaloids fraction extracted and purified have a significant attenuation in arsenic trioxide toxicity. The detoxification of total alkaloids extract was equal to that of N. domestica extract. Furthermore, apomorphine alkaloids, such as nantenine and domesticine, can be used as index components to establish a quality control method for total alkaloids.

Histomorphological Identification of Nandina domestica / 中国实验方剂学杂志

Objective::Histomorphological study of the Nandinae Radix, Nandinae Caulis, Nandinae Folium, Nandinae Fructus were conducted to provide the basis for the identification of its authenticity and falsehood. Method::The origin and macroscopic identification were used to describe the plant morphology and the appearance characteristics of all medicinal parts. The microscopic characteristics of the medicinal parts, such as roots, stems, leaves, flowers, fruits, were observed and photographed by paraffin section and powder preparation techniques. Result::It was found that the morphological characteristics of the original plant were consistent with the descriptions in herbaceous books. There was no pith on the cross-section of roots. In the transverse section of stems, there were intermittent circular fiber bundles in the cortex and a cap-shaped fibrous bundle in the inner part of the xylem. In the transverse section of the leaves, the palisade tissue was wider and the fiber bundles around the main vascular bundles formed a ring. In the transverse section of petiole, the fiber bundles were arranged intermittently into rings. In the transverse section of fruit, multi-layered sclereids formed a ring in the mesocarp. The powder characteristics of root and stem mainly contained crystalliferous sclereids. There were crystal sheath fibers and stomatal infinitive in the leaf powder, and pollen grains in the flower powder, with 3-hole grooves, obvious reticulate engraving pattern in the outer wall and more reticulate cells. There were a large number of branched sclereids and calcium oxalate square crystals in the fruit powder. Conclusion::The above-mentioned morphological and microscopic features have identification significance, and provide scientific basis for the authenticity identification, the quality standard and the utilization of resources of Nandina domestica.

Research Progress of Houpo Wenzhongtang / 中国实验方剂学杂志

Houpo Wenzhongtang was contained in Neiwaishang bianhuolun.It was composed of seven medicinal materials of Magnoliae Officinalis Cortex(processed with ginger),Citri Reticulatae Pericarpium(remove white sac),Glycyrrhizea Radix et Rhizoma(processed with honey),Alpiniae Katsumadai Semen,Poria,Aucklandiae Radix and Zingiberis Rhizoma.It was a classic prescription for treatment of deficient cold of spleen and stomach,distention of chest and abdomen,autumn and winter guest cold crime stomach and feel pain at times by LI Dongyuan,who was a famous doctors in Jin-Yuan dynasties.It has been included in the Catalogue of Ancient Classical Prescription(The First Batch) issued by the State Administration of Traditional Chinese Medicine in 2018.This paper systematically reviewed the relevant research progress of Houpo Wenzhongtang from the aspects of famous doctors' theory,usage and dosage,chemical composition,quality analysis,pharmacology and toxicology,clinical application in database,and to provide a reference for further exerting the clinical application of this classical prescription.

Phenolic Components from Roots of Psidium guajava / 中国实验方剂学杂志

Objective:The different parts of Psidium guajava(Myrrhinaceae) have different bioactivities. There are intensive studies for chemical constituents of its leaves and fruits at present. However,there are a few studies for the roots. Therefore, we systematically investigated the chemical constituents of phenolic acids in roots of P.guajava. Method:The chemical constituents were isolated and purified by various column chromatography methods and semi-preparative HPLC. The isolated compounds were identified by physicochemical properties and spectral analysis. Result:Sixteen phenolic components were isolated from the ethanol extract and identified as 3,3',4'-tri-O-methylellagic acid(1),3,3',4-tri-O-methylellagic acid-4'-O-β-D-glucopyranoside(2),3-O-methylellagic acid-4'-O-α-L-rhamnopyranoside(3),3'-O-methyl-3,4-O,O-metheneellagic acid-4'-O-β-D-glucopyranoside(4),gallic acid(5),methyl gallate(6),ethyl gallate(7), 3,4,5-trimethoxypheny-1-β-D-glucopyranoside(8),3,5-dimethoxy-4-hydroxy-benzoic acid-7-O-β-D-glucoside(9),1-hydroxy-3,4,5-trimethoxyphenyl-1-O-[6'-O-(4″-carboxy-1″,3″,5″-trihydroxy)phenyl]-β-D-glucopyranoside(10),vanillic acid(11),protocatechuic acid(12),secoisolariciresinol 9-O-β-D-glucopyranoside(13),phloretin 4'-O-β-D-glucopyranoside(14),cinchonain Ib(15) and epicatechin(16). Conclusion:Compounds 3,4,6,8-10,13-15 were isolated from this plant for the first time.

Advances in CRISPR/Cas9 gene-editing system / 基础医学与临床

CRISPR/Cas9 gene-editing system has been broadly used in various fields of bioscience and medicine in recent years. The system can be guided by RNA to specific DNA site thus achieving targeted gene editing.Off-target effect and editing efficiency remain to be two crucial challeges to the system. Currently, a number of researches have been focused on the optimization of the system by reducing off-target effects and increasing editing efficiency, which may enhance its safety and expand its application.

Recombinant Expression and Purification of Mouse Nectin-like 4 Glycoprotein in 293ET Cell Line / 中国医学科学杂志(英文版)

Objective To screen the transient and stable cell lines with high production of Nectin-like 4 (Necl-4) protein. Methods First, cDNA sequences encoding the extracellular domain of Necls were cloned into the modified vector pAPtag at the N terminus of alkaline phosphatase (AP) for fusion expression. Next, 293ET cells stably expressed Necls-AP fusion protein and secreted it into the culture medium which were detected by the AP activity assay and Western blot analysis. Then, by adding N-glycosylation processing inhibitor kifunensine into the medium, complex glycan was inhibited to generate. The residual glycan of purified protein was removed by endoglycosidase H. Finally, AP protein was removed by using human rhinovirus protease and size exclusion chromatography. The concentration of purified Necl-4 protein was monitored by measuring the absorbance at 280 nm and analyzed by SDS-PAGE. Result The transient and stable cell lines with high production of Necl-4 protein were screened by the color reaction with the AP-tag in the recombinant vector. The soluble and active form of purified Necl-4 protein was obtained after deglycosylation of native N-glycan protein with an expression level of 4 mg/L culture and purity of 95%. Conclusions By using modified AP mammalian protein expression system, we can easily screen the high productive stable cell lines by using AP activity assay. By adding mannosidase inhibitor kifunensine into the medium and cutting purified protein by using endoglycosidase H, we can obtain deglycosylated Necl-4 protein in milligram quantities. Our method might throw a light on the expression and purification of glycoprotein for structural and functional studies.

Visual and refractive outcomes of implantable collamer lens with a central hole for high myopia / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate the efficacy of implantable collamer lens (ICL)-V4c implantation in the correction for high myopia. METHODS: This retrospective analysis collected twenty-five patients (43 eyes) with high myopia treated with ICL/TICL implantation from September to December 2016 at Department of Ophthalmology, West China Hospital. Routine ophthalmic examinations were performed. The patients were followed 3mo to monitor uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), refractive error, intraocular pressure, corneal endothelial cell count, contrast sensitivity and scores for Quality of Life lmpact of Refractive Correction Questionnaire ( QIRC ). Statistical analyses were performed using repeated measures ANOVA, LSD-t test and paired-samples T test. RESULTS: Postoperative UCVA and BCVA were significantly higher than before surgery (all P<0. 01). Postoperative spherical mirrors at all time points were lower than before surgery(all P<0.01),and there was no significant difference between the different time point postoperative (all P>0. 05). The safety index in 3mo postoperatively was 1.38士0.24 and the effective index was 1. 22士0. 15. There was no statistical difference in intraocular pressure between preoperative and postoperative time points (F=1.464, P=0.215). There was no statistical difference in endothelial cell density at 3mo after surgery compared to preoperative (t=-0.544, P= 0. 586). The postoperative Log contrast sensitivity (LogCS) of 12.0 cycles per degree (c/d) and 18.0 c/d were significantly higher compared to the preoperative (F=4.347, P=0.010; F=4.539, P=0.005; respectively), but other (the postoperative LogCS of 3.0 c/d and 6.0 c/d) differences were statistically insignificant (F=1.094, P=0.354;F=1 325,P=0.271;respectively). There was no significant difference in the amount of change in the QIRC scores between preoperative and postoperative(F=2.669, P=0 094). CONCLUSION: ICL V4c implantation is a safe and effective method for the correction of high myopia.

Comparison of corneal thickness measured by CASIA SS-1000 OCT and Sirius anterior eye assay system / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To compare CASIA SS-1000 and Sirius OCT sweep anterior segment analyzer instrument for measuring normal corneal vertex thickness (CCT) and the thinnest corneal thickness (TCT) results the difference,correlation and consistency,and provide a theoretical basis for clinical application.METHODS:This was a prospective study.A total of 34 normal subjects were collected.The subjects were measured by the same skilled operator.The SS-1000 OCT was first used,and then the corneal thickness was measured repeatedly by Sirius anterior segment analyzer.Paired t test and Bland-Altman were used to evaluate the consistency of corneal apex and corneal thinnest point between SS-1000 OCT and Sirius anterior segment analyzer.RESULTS:The mean corneal apex measured by SS-1000 OCT and Sirius corneal topography were 517.62± 25.29μm and 518.47±27.23μm CCT,respectively.The thinnest points of SS-1000 OCT and Sirius anterior segment analyzer CCT were 513.53±25.06μ m and 515.32± 26.69μm,respectively.Paired t test showed that the difference on corneal thickness of vertex was not statistically significant (P＞0.05),but the thinnest corneal thickness was statistically significant (P＜ 0.05).Pearson analysis of the two devices,the correlation is 0.969,0.965.The results of 95％ consistency limiting analysis on the corneal vertex thickness by Bland-Altman was (-14.22μm,12.52μm),that of the thinnest corneal thickness was (-15.61μm,12.03μm),4％ (3/68) was out of the 95％ consistency limiting,but the thinnest corneal thickness was of a little larger differences.CONCLUSION:SS-1000 OCT measurement of CCT and Sirius anterior segment analyzer is highly consistent,in clinical work can be considered alternative,but the thinnest point of the cornea can not be replaced each other.

Two new compounds from Smilax glaucochina / 中国中药杂志

A new sesquiterpenes named glaucochinarol A (1) and a new phenylpropane glycoside named glcacochinaside A (2), together with six known ones, including trichothecolone (3), β-D-(6-O-trans-feruloyl)fructofuranosyl-α-D-O-glucopyranoisde (4), 3,4,5-trimethoxyphenyl-β-D-glucopyranoside (5), (4R)-p-menth-1-ene-7,8-diol-7-O-β-D-glucopyranoside (6), naringenin (7), and emodin-8-O-β-glucoside (8) were isolated from smilax glaucochina warb. Their structures were elucidated on the basis of NMR, MS and published data. Compounds 3-8 were isolated from the species for this first time.

A new triterpenoid from roots of Psidium guajava / 药学学报

To investigated the chemical constituents of the roots of Psidium guajava,we isolated seven compounds by silica gel column chromatography.These include oleanane derivatives,2 α,3β,6β,23-tetrahydroxylurs-12,20(30)-dien-28-oic acid β-D-glucopyranoside (1),2α,3β,6β,23-tetrahydroxylurs-12,18-dien-28-oic acid β-D-glucopyranoside (2),2α,3β,23-trihydroxylurs-12,18-dien-28-oic acid β-D-glucopyranoside (3), nigaichigoside F1(4),asiaticoside C (5),2α,3β,6β,19α,23-pentahydroxylurs-12,18-dien-28-oic acid β-Dglucopyranoside (6) and 2α,3β,19α,23-tetrahydroxylurs-12-en-28-oic acid (7).Their structures were elucidated on the basis of spectral analysis with reference to the published data.Compound 1 is brand new,compounds 2-6 were first isolated from this plant.The new compound was evaluated for their cytotoxic activity against human hepatoma Bel 7402 in vitro.The results were expressed as the ratio of inhibiting Bel 7402 cells growth by comparing to untreated cells.The new compound (concentration:25 μmol·L-1) showed the ratio values of 52.5%.

Studies on chemical constituents of rhizomes of Smilax trinervula / 中国中药杂志

The chemical constituents were separated and purified from the 70% ethanol extract of Smilax trinervulaby various chromatographic methods including silica gel, Sephadex LH-20, MCI and preparative HPLC. Their structures were obtained and identified by analysis of the spectroscopic data. Compounds 1-11 were separated from this genus for the first time. Compound 12 was obtained from S. trinervula for the first time.

Research progress on Acorus tatarinowii and its active ingredients in treatment of Alzheimer's disease / 中草药

Currently, Chinese materia medica is a research hotspot on the treatment of Alzheimer's disease (AD). The extracts from Acorus tatarinowii have a variety of pharmacological effects on the central nervous system, among which the volatile oil and β-asarone are the main active ingredients in A. tatarinowii reported on the effect on improving the learning and memory and prevention of AD. This review focuses on the main active ingredients in A. tatarinowii and progresses in their different therapeutic effects and proposed pharmacological mechanisms for AD on the basis of referring to the relevant literatures at home and abroad.

Flavonoids from leaves of Psidum littorale / 药学学报

We investigated the chemical constituents of the leaves of Psidum littorale, which include 16 flavonoids, including seven flavonols, six flavonoid glycosides and three flavonones. The compounds were isolated by silica gel column chromatography. Their structures were elucidated on the basis of spectral analysis and by comparison with published data. Seven flavonols were kaempferol (1), isorhamnetin (2), myricetin-3,7,3'-trimethyl ether (3), laricitrin (4), quercetin (5), myricetin (6) and quercein-3,4'-dimethyl ether (7), six flavonoid glycosides were guaijaverin (8), hyperoside (9), 5,4'-dyhydroxy-3,7,5'-methoxyflavone-3'-O-β-D-glucoside (10), laricitrin-3-O-xyloside (11), myricetin-3-O-α-L-rhamnopyranoside (12) and myricetin-3-O-β-D-xyloside (13). Three flavonones were 4'-O-methyldihydroquercetin (14), dihydroapigenin (15) and ampelopsin 4'-O-β-D-glucopyranoside (16). Compound 10 is a new chemical, compounds 2-4, 7, 10-16 were first isolated from this plant. 1H NMR and 13C NMR data of compound 11 were not reported in literature.

Quality control of Maca (Lepidium meyenii) / 中国中药杂志

To control the quality of Maca, the quality standard was established in this study. According to the methods recorded in the Appendix of Chinese Pharmacopoeia (2010 Edition), the water, extract, total ash, acid insoluble substance, and heavy metals inspections in Lepidium meyenii were carried out. N-benzyl-9Z, 12Z-octadecadienamide in L. meyenii was identified by TLC, and it was determined by HPLC. The results showed that the N-benzyl-9Z, 12Z-octadecadienamide identification of TLC was a strong mark and specificity. In content determination experiment, the linearity of N-benzyl-9Z, 12Z-octadecadienamide was in the range of 0.01-2 microg (r = 0.9998), and the average recovery (n=9) was 99.27% (RSD 2.0%). The methods were simple, accurate, with good reproducibility. It is suitable for quality control L. meyenii.

Research progress of bioactivity of steroidal saponins in recent ten years / 中国中药杂志

Steroidal saponins have a wide range of pharmacological effects and biological activities, such as anti-tumor, antifungal, hypoglycemic, immune regulation, insecticides, etc. In the last ten years, some new structures of steroidal saponins compounds were found from natural plants, they have some new and different activities. In order to accelerate the research on the drug innovation of steroidal saponins, we summarized the new progress of the research on such compounds.

The application of respiratory exerciser tri-ball in pulmonary rehabilitation patients with chronic obstructive pulmonary disease / 重庆医学

Objective To discuss the function of respiratory exerciser tri‐ball in pulmonary rehabilitation (PR) patients with chronic obstructive pulmonary disease(COPD) .Methods Prospectie case‐control study was used in COPD patients ,the patients were randomly divided into three groups ,60 COPD patients (group A) using respiratory exerciser TRI‐BALL ,58 COPD patients (group B) using traditional pursed lips ventral breathing training ,and 58 COPD patients(group C) using general internal medicine treatment .Results Group A :compared with before breath training ,the increases of FEV1/FEV1 predicted (% ) and MVV/MVV predicted (% ) and the decrease of quality of life score (QOL) were statistically significant after breath training (P0 .05) .Group B :compared with before breath training ,the decrease of QOL was statistically significant (P0 .05) .Compared with control group after breath training ,the increases of FEV1/FEV1 predicted (% ) and MVV/MVV predicted (% ) and the decrease of quality of life score (QOL) were statistically significant in group A (P0 .05) .Compared the changes of pulmonary function test(PFT) index and QOL between group A and B ,the increments of FEV1/FEV1 predicted (% ) and MVV/MVV predicted (% ) were statistically significant in group A(P0 .05) .Conclusion It is useful to improve the pulmonary function and quality of life in patients with COPD using respiratory exerciser tri‐ball .It is more effective than traditional pursed lips ventral breathing training ,due to the equipment is very small ,cheap ,easy to quantify training and convenient for household use ,it is worth to be popularized in primary hospital .

A Case of Lemierre's Syndrome in Young Patient with Acute Peritonsillar Abscess

Lemierre's syndrome is very rare and is characterized by previous oropharyngeal infections, such as pharyngitis or peritonsillar abscess, leading to high fever, internal jugular venous thrombosis, and metastatic infections to multiple internal organs. Prompt and accurate diagnosis followed by early treatment with antibiotics is very important because its mortality is high if treatment is delayed. We report on the case of a 23-year-old female who was transferred to our hospital with a left peritonsillar abscess combined with left jugular venous thrombosis, complaining of a sore throat and left submandibular swelling, and diagnosed as Lemierre's syndrome. Finally, she was treated successfully without anticoagulants.