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Determining the functional consequences of karyotypic changes is invariably challenging because evolution tends to obscure many of its own footprints, such as accumulated mutations, recombination events, and demographic perturbations. Here, we describe the assembly of a chromosome-level reference genome of the gayal (Bos frontalis) thereby revealing the structure, at base-pair-level resolution, of a telo/acrocentric-to-telo/acrocentric Robertsonian translocation (2;28) (T/A-to-T/A rob[2;28]). The absence of any reduction in the recombination rate or genetic introgression within the fusion region of gayal served to challenge the long-standing view of a role for fusion-induced meiotic dysfunction in speciation. The disproportionate increase noted in the distant interactions across pro-chr2 and pro-chr28, and the change in open-chromatin accessibility following rob(2;28), may, however, have led to the various gene expression irregularities observed in the gayal. Indeed, we found that many muscle-related genes, located synthetically on pro-chr2 and pro-chr28, exhibited significant changes in expression. This, combined with genome-scale structural variants and expression alterations in genes involved in myofibril composition, may have driven the rapid sarcomere adaptation of gayal to its rugged mountain habitat. Our findings not only suggest that large-scale chromosomal changes can lead to alterations in genome-level expression, thereby promoting both adaptation and speciation, but also illuminate novel avenues for studying the relationship between karyotype evolution and speciation., (© The Author(s) 2023. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.)

Objective: Osteosarcoma is the third most frequently diagnosed cancer among adolescents. Immunotherapy is an effective curative treatment for metastatic osteosarcoma patients. This study aimed to further reveal the significance of metabolism in tumor progression, and to categorize molecular subtypes for guiding personalized therapy., Materials and Methods: Univariate Cox regression analysis was performed to screen metabolism-related genes associated with osteosarcoma prognosis. A molecular subtyping system was developed by unsupervised consensus clustering. Survival analysis and functional analysis were used to evaluate the performance of subtyping and characterize the TME of subtypes. Stepwise Akaike information criterion (stepAIC) was employed to optimize the prognostic model., Results: C1 and C2 subtypes showed distinct prognosis, with more favorable survival in C2 subtype. C2 subtype presented a higher immune infiltration and active anti-tumor response. Notably, C2 subtype was predicted to have better immune response to immune checkpoint blockade. In addition, a 5-gene prognostic signature with robust ability to classify patients into high-risk and low-risk groups was developed., Conclusions: The study revealed the critical role of metabolism in tumorigenesis by comparing the features between the two subtypes. Oncogenic pathways including epithelial mesenchymal transition (EMT), glycolysis and hypoxia may be closely involved in the correlation with metabolism. Importantly, we developed a novel subtyping system and a 5-gene signature with high potential to be applied in clinical practice.

Lithium-free anode dual-ion batteries have attracted extensive studies due to their simple configuration, reduced cost, high safety and enhanced energy density. For the first time, a novel Li-free DIB based on a carbon paper anode (Li-free CGDIB) is reported in this paper. Carbon paper anodes usually have limited application in DIBs due to their poor electrochemical performance. Herein, by using a lithium bis(fluorosulfonyl)imide (LiFSI)-containing electrolyte, the battery shows outstanding electrochemical performance with a capacity retention of 96% after 300 cycles at 2C with a stable 98% coulombic efficiency and 89% capacity retention after 500 cycles at 5C with a stable coulombic efficiency of 98.5%. Moreover, the electrochemical properties of the CGDIB were investigated with a variety of in situ characterization techniques, such as in situ EIS, XRD and online differential electrochemical mass spectrometry (OEMS). The multifunctional effect of the LiFSI additive on the electrochemical properties of the Li-free CGDIB was also systematically analyzed, including generating a LiF-rich interfacial film, prohibiting Li dendrite growth effectively and forming a defective structure of graphite layers. This design strategy and fundamental analysis show great potential and lay a theoretical foundation for facilitating the further development of DIBs with high energy density., Competing Interests: The authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)

Syngnathids (seahorses, pipefishes and seadragons) exhibit an array of morphological innovations including loss of pelvic fins, a toothless tubular mouth and male pregnancy. They comprise two subfamilies: Syngnathinae and Nerophinae. Genomes of three Syngnathinae members have been analyzed previously. In this study, we have sequenced the genome of a Nerophinae member, the Manado pipefish ( Microphis manadensis ), which has a semi-enclosed brood pouch. Comparative genomic analysis revealed that the molecular evolutionary rate of the four syngnathids is higher than that of other teleosts. The loss of all but one P/Q-rich SCPP gene in the syngnathids suggests a role for the lost genes in dentin and enameloid formation in teleosts. Genome-wide comparison identified a set of 118 genes with parallel identical amino acid substitutions in syngnathids and placental mammals. Association of some of these genes with placental and embryonic development in mammals suggests a role for them in syngnathid pregnancy., (© The Author(s) 2020. Published by Oxford University Press on behalf of China Science Publishing & Media Ltd.)

Lithium (Li) metal is a favorable anode for most energy storage equipment, thanks to its higher theoretical specific capacity. However, nonuniform Li nucleation/growth results in large-sized and irregular dendrites generated from the Li anode, which causes rapid capacity fade and serious safety hazard, hindering its widespread practical applications. In this paper, with the aid of a lithium nitrate (LiNO 3 ) additive in a carbonate-based electrolyte, the Li anode shows low hysteresis for in excess of 1000 h at a current density of 0.5 mA cm -2 . At the same time, a Li-graphite dual-ion battery exhibits an outstanding cycling stability at 5C; after 1000 cycles, 81% of the capacity is retained. After calculation, the Li-graphite dual-ion battery shows a competitive specific energy density of 243 Wh kg -1 at a power density of 234 W kg -1 . Moreover, the linear sweep voltammetry test was first performed to analyze the Li nucleation/growth mechanism and explain the effect of the LiNO 3 additive. The superior electrochemical properties of the Li-graphite dual-ion battery are ascribed to the formation of smooth Li composed of Li nanoparticles and a steady solid electrolyte interface film.

Background: Pistachio (Pistacia vera), one of the most important commercial nut crops worldwide, is highly adaptable to abiotic stresses and is tolerant to drought and salt stresses., Results: Here, we provide a draft de novo genome of pistachio as well as large-scale genome resequencing. Comparative genomic analyses reveal stress adaptation of pistachio is likely attributable to the expanded cytochrome P450 and chitinase gene families. Particularly, a comparative transcriptomic analysis shows that the jasmonic acid (JA) biosynthetic pathway plays an important role in salt tolerance in pistachio. Moreover, we resequence 93 cultivars and 14 wild P. vera genomes and 35 closely related wild Pistacia genomes, to provide insights into population structure, genetic diversity, and domestication. We find that frequent genetic admixture occurred among the different wild Pistacia species. Comparative population genomic analyses reveal that pistachio was domesticated about 8000 years ago and suggest that key genes for domestication related to tree and seed size experienced artificial selection., Conclusions: Our study provides insight into genetic underpinning of local adaptation and domestication of pistachio. The Pistacia genome sequences should facilitate future studies to understand the genetic basis of agronomically and environmentally related traits of desert crops.

Dual-ion battery complements lithium-ion batteries in terms of the use of inexpensive materials and ease to construct cells. To improve the safety and energy density of dual-ion battery, in this paper, a novel MnO-graphite dual-ion battery is reported for the first time. Microporous MnO materials are used as anode, which exhibits a low conversion potential and a low voltage hysteresis. The MnO-graphite dual-ion battery can deliver a capacity of 104 mAh g -1 at 0.5C and exhibits good rate performances and cycling stability (capacity retention >93% after 300 cycles). A mechanism is proposed to explain the irreversibility in capacity during the initial cycle by using operando X-ray diffraction in combination with online electrochemical mass spectrometry and electrochemical impedance spectroscopy.

Aims: Protocatechuic acid (PCA) is a phenolic compound found in many antiviral Chinese herbal medicines. HNF4α and HNF1α, the members of hepatocyte nuclear factor (HNF) family, play an important regulatory role in the gene transcription of hepatitis B virus (HBV). Previous studies found that PCA inhibited HBV antigen secretion and HBV DNA replication in HepG2.2.15 cells, but its anti-HBV mechanism has not been fully understood. We aim to illustrate the anti-HBV mechanism of PCA., Materials and Methods: MTT was used to estimate cytotoxicity. The content of HBsAg or HBeAg was detected using an enzyme-linked immunosorbent assay kit. HBV DNA in cell-free culture media was detected by PCR kit. HNF1α and HNF4α mRNA expression was detected by real-time PCR. HNF1α, HNF4α and ERK1/2 protein expression was detected by western blotting and HBV promoter activity was tested by luciferase reporter assay., Key Findings: Our results demonstrated that PCA inhibited the gene transcription and protein translation of HNF1α and HNF4α in Huh7 and HepG2.2.15 cells, as well as the promoter activities of HBV X and preS1 in Huh7 cells transfected with the luciferase reporter plasmid of HBV promoter. Further study suggested that PCA induced the phosphorylation of extracellular-signal-related kinase (ERK) 1/2, and thereby inhibited HNF4α and HNF1α expression in HepG2.2.15 cells to exert its antiviral activity., Significance: To our knowledge, this study is the first to reveal the anti-HBV mechanism of PCA. Our results demonstrate that PCA inhibits HBV replication by activating ERK1/2 pathway and subsequently down-regulating HNF4α and HNF1α in HepG2.2.15 cells., (Copyright © 2017 Elsevier Inc. All rights reserved.)

In this study, an HPLC method was developed for simultaneous determination of seven alkaloids (cytosine, oxymatrine, N-oxysophocarpine, N-methylcytisine, sophoranol, matrine, and sophocarpine) and three flavonoids (trifolirhizin, fermononetin, and maackiain) from different samples of Sophorae Tonkinensis Radix et Rhizoma. Samples were analyzed on a Welch XtimateTM C₁₈ column (4. 6 mm× 250 mm, 5 μm) eluted with the mobile phase of acetonitrile (A) and 0.01 mol•L⁻¹ ammonium acetate solution (pH 8.0) (B) in a linear gradient mode as follows: 0-20 min,4%-14% A;20-30 min,14%-25% A;30-45 min,25%-40% A;45-65 min,40%-55% A;65-75 min,55% A. The flow rate of the mobile phase, the column temperature, and the PDA detector wavelength were set at 1.0 mL•min⁻¹, 30 ℃, and 225 nm, respectively. For the method validation, these ten compounds showed good separation and satisfactory linearity (r≥0.999 7) within the concentration ranges tested. The mean recoveries were in the range of 98.60% to 102.6% with the RSD (n=6) between 0.60% and 3.7%. This method was proved to be simple, accurate and repeatable. The quantitative results showed that there were significant differences in the contents of seven alkaloids and three flavonoids among the different samples. This result revealed that the quality of Sophorae Tonkinensis Radix et Rhizoma varied widely. This method could be used for the simultaneous determination of the multi-ingredients from Sophorae Tonkinensis Radix et Rhizoma, which might provide scientific evidences to evaluate/control the quality of Sophorae Tonkinensis Radix et Rhizoma, comprehensively., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

We prepared in this work an anchoring porphyrin and a series of hat-porphyrins. The zinc atom of the hat-porphyrins can be coordinated axially with the pyridine moiety of the anchoring porphyrin which is anchored on the titania surface by a carboxyl group. The structures of the assemblies were confirmed using computational calculations, transmission electron microscopy (TEM) and energy dispersive spectrometry (EDS). Solar cell devices of the monomer anchoring porphyrin and its assemblies were fabricated and the photovoltaic performances were measured under standard AM 1.5 sunlight irradiance. We found that the assembly devices showed higher JSC and lower VOC than that of the monomer anchoring porphyrin device. However, the comprehensive influence of JSC and VOC led to an enhancement in the solar-to-electric power-conversion efficiency (PCE) of the assemblies. We also studied the variation of JSC and VOC using electronic absorption and emission spectroscopy, charge extraction measurements, transient photovoltage decay measurements and electrochemical impedance spectroscopy.

Currently, chemotherapy is one of the main therapy for cancer. But the traditional antitumor drugs are systemic distribution in vivo, they are difficult to achieve an effective drug concentration in the tumor tissue and don't have the ability to distinguish normal cells and tumor cells by themselves, that cause systemic toxicity easily and can not meet the clinical needs. With the research on mesoporous silica nanoparticles (MSNs) deepening, more and more attention in the drug delivery system have been payed to in recent years, because of its unique physicochemical structure characteristics, it has the effect on specific targets, directly inhibits the tumor cell growth, reduces the side effects to normal cells, tissues and organs and can be long-term medication, etc. It is expected to be excellent carriers of antitumor drugs. MSNs application in the field of cancer treatment has now become a hot research field of medicine. In this paper, the latest research about MSNs in antitumor drugs targeting delivery system from 2008 to 2015 is summarized, including the application of MSNs separately in antitumor drug targeting, passive targeting, active targeting, physical or chemical conditions response targeting and other compound targeting drug delivery system. We expect it to provide a reference to the toxicity reducing and efficacy enhancing and further development of chemical medicine, natural medicine and monomeric compound of chinese herbal medicine.

Objective: To establish molecular method for identification of Ilex plants using the internal transcribed spacer(ITS) bar- code., Methods: DNA was extracted using centrifugal columnar kits. The ITS sequences of all experimental samples were amplified and sequenced using a pair of universal primers. The alignment and the phylogenetic tree were performed in MEGA5. 0., Results: The length of ITS sequence in each species was 759 bp,in which there were 216 sites discrepancy., Conclusion: DNA barcoding technology based on ITS sequence can be used to identify plants of Ilex genus.

Triptolide (TP) is a major active component in Tripterygium root, but its therapeutic window was very narrow due to its severe multi-organ toxicity. In this work, the effect of TP combined with glycyrrhetic acid (GA) on mRNA expression and activity of four cytochrome P450 (CYP) enzymes in rat liver was studied after intragastric administration of TP (0.05, 0.3 and 0.6 mg x kg(-1) x day(-1)) and TP (0.6 mg x kg(-1) x day(-1)) combined with GA (30 mg x kg(-1) x day(-1)) for 7 consecutive days. Compared with the control, the high dose of TP significantly up-regulated the mRNA expression levels of CYP2E1, 1A2, 3A1 and 2C11, the co-administration of TP and GA further up-regulated the mRNA expression levels of CYP3A1, 2C11 and 2E1 as compared with the high dose of TP. Meanwhile, TP at high dose and combined with GA significantly increased CYP3A-associated testosterone 6beta-hydroxylation activity (2.2-fold and 4.1-fold, respectively) as compared with the control. Because TP is mainly metabolized by CYP3A2 in male rats, the present work indicated that TP-induced increase of CYP3A activity might be an important reason for the rapidly metabolic clearance of TP in rat liver, and GA can reduce the hepatotoxicity of TP by promoting its hepatic metabolic clearance. Furthermore, the results also suggest that the drug interactions might be occurred when TP and GA were co-administered with other CYP3A substrate drug.

Abstract: The activities of four CYP450 enzymes (CYP3A, 1A2, 2El and 2C) and the mRNA expression levels of CYP1A2, 2El, 2Cll and 3A1 in rat liver were determined after Wistar rats were orally administered with brucine (BR) at three dosage levels (3, 15 and 60 mg.kg-1 per day) and the high dose of BR combined with glycyrrhetinic acid (GA, 25 mg.kg-1 per day) or liquiritin (LQ, 20 mg.kg-1 per day) for 7 consecutive days. Compared with the control, brucine caused 24.5% and 34.6% decrease of CYP3A-associated testosterone 6beta-hydroxylation (6betaTesto-OH) and CYP2C-associated tolbutamide hydroxylation (Tol-OH), respectively, and 146.1% increase of CYP2El-associated para-nitrophenol hydroxylation (PNP-OH) at the high dose level. On the other hand, (BR+GA) caused 51.4% and 33.5% decrease, respectively, of CYP2El-associated PNP-OH and CYP1A2-associated ethoxyresorufin-O-de-ethylation (EROD) as compared with the high dose of BR group. Meanwhile, (BR+LQ) caused 41.1% decrease of CYP2El-associated PNP-OH and 37.7% increase of CYP2C-associated Tol-OH. The results indicated that the co-administration of BR with GA or LQ had effect on mRNA expression and activities of the CYP450 enzymes mentioned above to some extent, and the in vivo antagonism of LQ on BR-induced CYPs adverse effects and the in vivo inhibitory action of GA on CYP2E1 and 1A2 might play an important role in the detoxification of Radix Glycyrrhizae against Strychnos nux-vomica L.

To study the influence of the compatibility of ophiopogonis tuber and Chinese magnoliavine fruit with gastrodia rhizome on the pharmacokinetics of gastrodin in rat, three dosages of compound Tianma granule extract (equivalent to gastrodin 50, 100, 200 mg x kg(-1)) and one dosage of Tianma extract (equivalent to gastrodin 100 mg x kg(-1) were administered to rats by intragastric administration separately. Plasma samples were collected at different times and treated with methanol and acetonitrile to precipitate protein. The contents of gastrodin in plasma were determined by HPLC method. The mean plasma concentration-time curves of different medication administration teams were processed with WinNonlin 5.2.1 pharmacokinetic software. The pharmacokinetic parameters of different medication administration teams were analyzed with SPSS statistics 17.0 software. The results indicated that the in vivo kinetic process of gastrodin was fitted to first-order absorption un-compartment model at low, middle dosages and zero-order absorption un-compartment model at high dosage of compound Tianma granule extract. By comparison with the pharmacokinetic parameters of gastrodin (100 mg x kg(-1)) in Tianma extract, the significant decrease for Cmax and significant increase for MRT0-infinity in compound Tianma granule extract indicated that the compatibility of ophiopogonis tuber and Chinese magnoliavine fruit with Gastrodia rhizome can delay the absorption, reduce the elimination rate and prolong the action time of gastrodin in vivo.

To describe a modified simple iris suture for pupillary dilation technique during vitrectomy in cases with a miotic pupil. Four translimbal incisions were created with a sharp straight blade at 1:30, 10:30, 4:30, and 7:30 o'clock, respectively. The straight needle of 10-0 polypropylene suture and a Sinskey IOL hook was used to displace the pupillary margin toward the limbus. In 3 cases, four sutures caused a 6-mm to 9-mm square-shaped pupil, and the pupil was allowed to return to a smaller size at the end of the operation. It is simple and may reduce postoperative complications.

After oral administration of low, middle, high dose of simulative Maijunan tablets to SD rats and puerarin, hydrochlorothiazide at middle dosage to SD rats separately, plasma samples were collected at different times and treated with acetonitrile to precipitate protein. The contents of puerarin and hydrochlorothiazide in plasma were determined by HPLC method. The mean plasma concentration-time profiles of puerarin and hydrochlorothiazide at different dosages of medication administration were processed by 3P97 pharmacokinetic software and SPSS statistics 17.0 software. The results indicated that the in vivo kinetic processes of puerarin in rats were all fitted to a two-compartment open model and hydrochlorothiazide fitted to a one-compartment open model. Hydrochlorothiazide in vivo kinetic process in rats was in accordance with the linear dynamics. The combination of hydrochlorothiazide and rhynchophylla with pueraria promoted the absorption, reduced the elimination rate and prolonged the action time of puerarin in vivo. Meanwhile, the combination also promoted the absorption rate and the bioavailability, prolonged the action time and the accumulation time of hydrochlorothiazide in vivo.

Objectives: Despite its important therapeutic value, the metabolism of palmatine is not yet clear. Our objective was to investigate its in-vivo and in-vitro metabolism., Methods: Liquid chromatography-tandem electrospray ionization mass spectrometry (LC-ESI/MSn) was employed in this work. In-vivo samples, including faeces, urine and plasma of rats, were collected after oral administration of palmatine (20 mg/kg) to rats. In-vitro samples were prepared by incubating palmatine with intestinal flora and liver microsome of rats, respectively. All the samples were purified via a C18 solid-phase extraction procedure, then chromatographically separated by a reverse-phase C18 column with methanol-formic acid aqueous solution (pH 3.5, 70:30 v/v) as mobile phase, and detected by an on-line MSn detector. The structure of each metabolite was elucidated by comparing its molecular weight, retention time and full-scan MSn spectra with those of the parent drug., Key Findings: The results revealed that 12 metabolites were present in rat faeces, 13 metabolites in rat urine, 7 metabolites in rat plasma, 10 metabolites in rat intestinal flora and 9 metabolites in rat liver microsomes. Except for six of the metabolites in rat urine, the other in-vivo and in-vitro metabolites were reported for the first time., Conclusions: Seven new metabolites of palmatine (tri-hydroxyl palmatine, di-demethoxyl palmatine, tri-demethyl palmatine, mono-demethoxyl dehydrogen palmatine, di-demethoxyl dehydrogen palmatine, mono-demethyl dehydrogen palmatine, tri-demethyl dehydrogen palmatine) were reported in this work.

Kunming mice (male, weight 20 +/- 2g) were daily intragastric administration of Chinese nutgall extract (0.2 mL/10 g body weight, equal to 8 g Chinese nutgall material/1 kg body weight) for 30 days. Liver tissue mRNA were extracted from normal control group and Chinese nutgall treated group respectively, then were reversely transcribed to cDNA with dUTP labeled by different fluorescence (Cy3, Cy5) as hybridization probes. Both of the cDNA probes were mixed equally in 20 microL of hybridization solution and hybridized with mice complete genome oligonucleotide microarray. The fluorescent signals were acquired by laser scanner and analyzed by GenePix Pro 4.0 software. The biological function analysis and the pathway analysis of differentially expressed genes were performed according to Gene Ontology database. As a result, there were 461 genes differentially expressed in Chinese nutgall treated group, in which 373 genes were function-known and the others were function-unknown. Among the 461 genes, 267 genes were up-regulated and the others were down-regulated. The differentially expressed genes were involved in metabolism, DNA binding and transcription, protein synthesis and modification, cell cytoskeleton and cell adhesion, cell cycle and differentiation, ion channels and transporters, signal transduction, immune response and apoptosis of liver cell. The presented work might be quite important for understanding the pathogenic mechanisms of liver injury induced by Chinese nutgall.

In this work, liquid chromatography-electrospray ionization tandem ion-trap mass spectrometry (LC-MS(n)) was used to investigate the in-vivo and in-vitro metabolism of tectoridin. After oral administration of a single dose (100 mg kg(-1)) of tectoridin to healthy rats, faeces and urine samples were collected for 0-48 h and 0-24 h, respectively. Tectoridin was also incubated with rat intestinal flora and rat liver microsomes. Samples from in-vivo and in-vitro metabolism studies were purified using a C(18) solid-phase extraction cartridge, then separated using a reverse-phase C(18) column with methanol/ water (30:70, v/v, adjusted to pH 10.0 with ammonia water) as mobile phase and detected by an on-line MS(n) system. The structure of the metabolites was elucidated by comparing their molecular weights, retention times and full-scan MS(n) spectra with those of the parent drug. The results revealed six metabolites of tectoridin in urine (tectorigenin, hydrogenated tectorigenin, mono-hydroxylated tectorigenin, di-hydroxylated tectorigenin, glucuronide-conjugated tectorigenin and sulfate-conjugated tectorigenin); three metabolites in faeces (tectorigenin, di-hydroxylated tectorigenin and sulfateconjugated tectorigenin); one metabolite in the intestinal flora incubation mixture (tectorigenin), and four in the liver microsomal incubation mixture (tectorigenin, hydrogenated tectorigenin, mono-hydroxylated tectorigenin and di-hydroxylated tectorigenin). Except for tectorigenin, all other metabolites of tectoridin are reported for the first time.

The albumin is the richest protein in blood circulatory system, which can combine with many drugs and play an important role in transporing protein. In the present work, the non-covalent interaction between human serum albumin and cinnamic acid was studied by using fluorescence quenching method. The results showed that cinnamic acid had a powerful ability to quench the fluorescence of human serum albumin at excitation and emission wavelengths of lambda(ex) = 286 nm and lambda(ex) = 340 nm, respectively, in the reaction solution of pH 7.4. The binding constants(K) at 37 and 47 degrees C were found to be 1.276 7 x 10(3) and 3.404 1 x 10(3) L x mol(-1), with the number of binding site(n) of 0.758 6 and 0.835 6, respectively, suggesting that the reaction temperature is advantageous for the binding reaction in a way. The changes in the thermodynamic parameters of binding interaction at 37 and 47 degrees C indicated that the main binding force between cinnamic acid and human serum albumin was hydrophobic force. These provide important information for studying the pharmacological effects of cinnamic acid and the influence of cinnamic acid on the configuration change of HSA.

Amygdalin and its metabolites in rat urine were identified using liquid chromatography-electrospray ionization (ESI) tandem ion-trap mass spectrometry. The purified rat urine sample was separated using a reversed-phase C18 column with 10 mM sodium phosphate buffer (pH 3.1) containing 30% methanol as the mobile phase, amygdalin and its metabolites were detected by on-line mass detector in selected ion monitoring (SIM) mode. The identification of the metabolites and elucidation of their structure were performed by comparing the changes in molecular masses (DeltaM), retention times and MS(2) spectral patterns of metabolites with those of parent drug. At least seven metabolites and the parent drug were found in rat urine after i.v. injection of 100 mg/kg doses of amygdalin. Among them, six metabolites were reported for the first time.

Aim: To analyze the gene expression profiles of mice livers injured by Leigongteng and explore the relationship between the differentially expressed genes and liver damage., Methods: The experimental mice were randomly divided into a control group and a liver-injured group in which the mice were administrated 33 mu gamma of triptolide/kg per day for 30 d. Liver mRNAs were extracted from animals in both groups and were reverse-transcribed to cDNA with dUTP labeled by different fluorescence (Cy3, Cy5) as hybridization probes. The mixed probes were hybridized with oligonucleotide microarray chips. The fluorescent signal results were acquired by scanner and analyzed with software., Results: Among the 35852 target genes, 29 genes were found to be significantly differentially expressed, with 20 genes up-regulated and 9 genes down-regulated. The reliability of the differentially expressed genes was validated by RT-PCR experiments of 5 randomly selected differentially expressed genes., Conclusion: Based on the biological functions of the differentially expressed genes, it is obvious that the occurrence and development of liver damage induced by Leigongteng in mice are highly associated with immune response, metabolism, apoptosis and the cell skeleton of liver cells. This might be important for elucidating the regulatory network of gene expression associated with liver damage and it may also be important for discovering the pathogenic mechanisms of liver damage induced by Leigongteng.

Objective: A new method for the simultaneous quantitative determination of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill using reversed phase high performance liquid chromatographic method had been developed., Method: The optimum chromatographic conditions were as follows: Agilent Zorbax SB - C18 column (4.6 mm 250 mm, 5 m), acetonitrile-H2O (containing 6 mmol L(-1) KH2PO4, pH 4.6) gradient elution; as a detection wavelength of 343 nm., Result: The calibration curves of geniposide, baicalin and berberine were linear at the ranges of 4.50-110.00, 5.00-153.00, 6.40-191.00 mg L(-1), respectively. The limits of detection of the method were 0.77 ng for geniposide, 1.53 ng for baicalin and 1.43 ng for berberine hydrochloride. The recoveries of the method were 104.44% (RSD 1.79% ) for geniposide, 96.98% (RSD 1.76%) for baicalin, 101.08% (RSD 3.1%) for berberine hydrochloride., Conclusion: This method had been successfully applied to determine the content of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill.

A rapid, sensitive and specific liquid chromatographic-electrospray ionization (ESI) tandem ion trap mass spectrometric method has been developed for identification of dauricine and its metabolites in rat urine. Six healthy rats were administrated a single dose (100 mg/kg) of dauricine by oral gavage. The urine were sampled from 0 to 24 h and purified by using a C18 solid-phase extraction (SPE) cartridge, then the purified urine samples were separated on a reversed-phase C18 column using methanol/2 mmol/L ammonium acetate (70:30, v/v, adjusted to pH 3.5 with formic acid) as mobile phase and detected by an on-line MS detector. Identification and structural elucidation of the metabolites were performed by comparing their changes in molecular mass (Deltam) and full scan MS(n) spectra with those of the parent drug. At least eight metabolites (such as N-demethyl, dehydrogenate, demethoxyl, hydroxyl, glucuronide conjugated and sulfate conjugated metabolites) and the parent drug were found in rat urine.

A new MS-titration method for the non-covalent binding of protein-ligand based on the research of berberine and alpha1-acid glycoprotein was established. The major presumption of new method is that the total concentration of protein-ligand complex is approximately the same as the total concentration of acting protein if a certain extent of affinity is existed between protein and ligand, in addition, the mole amount of acting ligand is more than that of acting protein. The non-covalent binding behaviours between berberine and alpha1-acid glycoprotein was studied by using electrospray ionization ion trap mass spectrometry (ESI-ITMS) , and the results were verified by fluorescence quenching method. The results showed that the binding behaviours between berberine and alpha1-acid glycoprotein, for example, stability constant, number of binding site, type of the main binding force, were almost the same by using the new MS-titration method and fluorescence quenching method. Comparing with the reported MS-titration method, the presented MS-titration method in this paper is more simple and applicable, does not demand much for the devices, and can lead to reliable results in same cases.

Pectinases are mainly used in the food industry to clarify fruit juices and wine, improve oil extraction, remove the peel from the citrus fruit, increase the firmness of some fruits and degum fibres. The filamentous fungus Aspergillus oryzae, used for the production of traditional fermented foods, only could produce less pectinases under general conditions. So far only a few of PGs expressed in yeast or E. coli were reported but they did not show higher activity. The cDNA of mature PGA (without signal peptide) was synthesized with specific primers from total RNA of Aspergillus oryzae by RT-PCR. PGA cDNA was ligated into pET-28a( + ) expression vector, creating plasmid pET-28a( + )-pgA. The plasmid pET-28a( + )-pgA was transformed into E. coli Turner (DE3) plac I cells to express PGA heterogeneously. For improving the efficiency of PGA expression in E. coli, the conditions for expression of the PGA in E. coli were optimized. E. coli Turner (DE3) plac I cells with pET-28a( + )-pgA was first cultivated at 37 degrees, 220r/min until OD600nm reached about 0.8. Then, cultivation broth was added with 0.5 mmol/L IPTG and incubated at 15 degrees C, 170r/min for other 24 h for induced-expression of PGA. Our data showed that the activity of recombinant expressed PGA could reach to 70u/mL medium, which is 87.5-fold of the activity of PGA produced in culture of A. oryzae and superior than known recombinant expression amount of PGA reported by other researchers.

Aim: To establish a LC-MS(n) method for the identification of anisodamine and its metabolites in rat feces., Methods: Feces samples were collected after single administration of 25 mg x kg(-1) anisodamine to rats, and dipped in water for 1 h. Samples were then extracted by ethyl acetate. The pretreated samples were separated on a reversed-phase C18 column using a mobile phase of methanol / 0.01% triethylamine (adjusted to pH 3.5 with formic acid) (60 : 40, v/v) and detected by LC-MS". Identification of the metabolites and elucidation of their structures were performed by comparing their changes in molecular masses (deltaM), retention-times and full scan MS(n) spectra with those of the parent drug and blank feces., Results: The parent drug and its seven metabolites (6beta-hydroxytropine, nor-6beta-hydroxytropine, aponoranisodamine, apoanisodamine, noranisodamine and hydroxyanisodamine, tropic acid) were found in rat feces., Conclusion: This method is sensitive, rapid, simple, effective, and suitable for the rapid identification of drug and its metabolites in biologic samples.

Aim: To identify the hydroxylate metabolites and its sulfate conjugates of daidzein in rat urine., Methods: Urine samples from 0 - 24 h were collected after single ig dose of 500 mg x kg(-1) daidzein to each of six rats. The urine samples were purified by SPE column (SPE C18) and analyzed with liquid chromatographic-tandem electrospray ionization ion trap mass spectrometry (LC-ESI/MS(n)) for potential metabolites., Results: Several new hydroxylate metabolites and its sulfate conjugates were found and identified in rat urine., Conclusion: LC-ESI/MS(n) is proved to be a simple, rapid, sensitive and specific technique for identification of the hydroxylate metabolites and its sulfate conjugates of daidzein in rat urine.

Aim: To identify the main metabolites of jatrorrhizine in rat urine., Methods: The rat urine samples were collected 0 - 72 h after ig 12 mg x kg(-1) jatrorrhizine, then the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by combining liquid chromatography and tandem electrospray ionization ion trap mass spectrometry (LC-ESI/ITMS(n)). Identification and structural elucidation of the metabolites were performed by comparing the changes in molecular masses, retention-times and full scan MS(n) spectra with those of the parent drug., Results: At least seven phase I metabolites (such as de-methyl, de-hydrogen and hydroxyl metabolites) and eleven phase II metabolites (such as glucuronide conjugates and methyl-conjugates) were identified in rat urine., Conclusion: The developed LC-ESI/ITMS(n) method is not only simple and rapid but also sensitive and specific for the identification of metabolites of jatrorrhizine in rat urine.

Aim: To identify anisodine and its metabolites in rat plasma after ingestion of anisodine by combining liquid chromatography and tandem mass spectrometry (LC-MS(n))., Methods: Plasma samples from rats after a single orally administration of 20 mg anisodine were added with methanol to precipitate protein. Then, it was analyzed by LC-MS(n). Identification and structural elucidation of the metabolites were performed by comparing their changes in molecular masses, retention-times and full scan MS(n) spectra with those of the parent drug and blank plasma., Results: The results revealed that the parent drug and its four metabolites (norscopine, scopine, hydroxyanisodine, N-oxide anisodine) existed in rat plasma., Conclusion: This method is sensitive, rapid, simple, and it is suitable for the rapid identification of drug and its metabolits.

Aim: To identify the main metabolites of stachydrine in rat., Methods: The ionization, cleavage and chromatographic characteristics of stachydrine were studied by using high-performance liquid chromatography-electrospray ionization ion trap tandem mass spectrometry (HPLC-ESI/MS) for the first time. These characteristics of stachydrine were used as the basis for the analyses of metabolites in rat urine. The 0 - 24 h urine samples of rats after ig 25 mg x kg(-1) stachydrine were collected and purified by using C10 solid-phase extraction cartridge, and then analyzed by HPLC-ESI/MS to identify stachydrine and its metabolites., Results: The parent drug (stachydrine), 6 phase I metabolites (N-demethyl, dehydrogenation, ring-oxidation) and 2 phase II metabolites (glycine conjugates of 2 ring-oxidation products) were identified existing in rat urine., Conclusion: The presented method was proved to be sensitive, rapid, high selective and specific for the identification of stachydrine and its metabolites in rat urine.

Aim: To establish a rapid and sensitive LC-MSn method for the identification of trigonelline and its main metabolites in rat urine., Methods: After optimizing the detection conditions of LC-MSn chromatography and mass spectrometry using trigonelline, its ionization and cleavage in ESI-MS and ESI-MSn modes were summarized, then serving as the basis for the metabolite analysis of trigonelline in rat urine. The 0-48 h urine samples of rats were collected after iv 8 mg x kg(-1) trigonelline, then, the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by LC-MSn., Results: The structures of trigonelline metabolites were elucidated according to the changes of the molecular weights of the metabolites (deltaM) and their cleavage pattern in ESI-ITMSn. As a result, two phase I metabolites and the parent drug were identified existing in rat urine, and two phase II metabolites were identified., Conclusion: The LC-MSn method is rapid and high sensitive and specific, it is suitable for the identification of trigonelline and its metabolites in rat urine.

Comparison between CCl4 liver injury model and immunity liver injury model was studied through physiological, biochemical and gene expression profile methods, and the pathological mechanism of them were discussed. There are significant changes about the physiological, biochemical parameters including ALT,AST between two model groups and normal control group. Among the 14112 target genes, 379 and 293 differentially expressed genes were screened out from the both two model groups respectively in gene expression profile experiments, including 105 common, differentially expressed genes. The results indicate that the two models of CCl4 and immunity have some similarities. However, the differences between them are also obvious. It is useful for the further investigation on the liver-injury mechanism and pathological process of the two different liver-injury models.

Aim: To estabilish a rapid and sensitive LC-ESI-ITMSn method for the identification of ephedrine and its main metabolites in rat urine., Methods: After optimizing the detection condition of LC-ESI-ITMSn chromatography and mass spectrometry by using a standard ephedrine, the ionization and cleavage rules of ephedrine in ESI-MS and ESI-MSn modes were summarized, and then serving as the basis for the metabolite analysis of ephedrine in rat urine. Rat urine samples of 0-48 h were collected after ig 10 mg x kg(-1) ephedrine, then the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by LC-ESI-ITMSn., Results: The structures of ephedrine metabolites were elucidated according to the changes of the molecular weights of the metabolites (deltaM) and their cleavage pattern in ESI-ITMSn. As a result, three phase I metabolites and the parent drug ephedrine were identified existing in rat urine, but no phase II metabolites were found., Conclusion: The LC-ESI-ITMSn method is rapid and highly sensitive and sepecific, it is suitable for the identification of ephedrine and its metabolites in rat urine.

Aim: To identify the main metabolites of oxymatrine (OMT) in rats., Methods: To optimize the conditions of LC/ESI-ITMS' chromatograms and spectra by oxymatrine and matrine (MT), and summarize their ionization and cleavage rules in ESIMS, then serving as the basis for the metabolite analyses of oxymatrine in rats. To collect the 0-24 h urine samples of the rats after ip 40 mg x kg(-1) oxymatrine, the samples were enriched and purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by LC/ESI-ITMS. The structures of OMT metabolites were identified according to their retention times and ESI-ITMSn rules., Results: Six phase I metabolites and the parent drug OMT were found in the rat urine, and the main metabolite was MT. No phase II metabolites were found., Conclusion: The developed LC/ESI-ITMSn methods to identify the metabolites of oxymatrine in rats is not only simple and rapid but also sensitive and specific. This technology is one of the most efficient methods for the analysis of drug metabolites.

(Tb1-x Tmx).L2.(ClO4).2H2O(x = 0.000 to 0.200, L = C6H5SOCH2COO-) have been synthesized. The coordination compounds have been studied by means of composition analysis, molar conductivity, IR, and the condition of coordination have been inferred. In the fluorescent spectra it was found that Tm3+ has a strongly sensitization effect to the fluorescence of Tb3+. The fluorescent emission intensity of Terbium perchlorate with phenylcarboxymethyl sulfoxide coordination compounds would be enhanced by Tm3+ in mixing. Tm3+ has a sensitization to the fluorescence of Tb3+ in the ratio of Tb3+:Tm3+ = 0.999:0.001-0.900:0.100. In the solubility experiment it was found that the complexes have high solubility in ethanol.

A high performance capillary electrophoresis method has been developed for separation and quantitative analysis of glibenclamide in Xiaotangling tablets. Electrophoretic conditions were as follows: micellar electrokinetic capillary chromatography (MECC) mode, trimethoprimum as internal standard, uncoated fused silica capillary (34.8 cm/39.5 cm(effective/total length), 50 microns i.d.), 25 mmol/L borax-30 mmol/L SDS(pH 9.0), applied voltage 17 kV, (+)-->(-), temperature 28 degrees C, detection wavelength 228 nm, pressure injection 68.95 kPa.s. Glibenclamide was separated successfully from other components within 14 minutes under the optimum conditions, and the relative peak area of glibenclamide increased linearly with the increase of its concentration within the range of 25 mg/L-275 mg/L. The recovery was (100.6 +/- 1.4)%. The method is simple, rapid and well reproducible, and can be used as a reliable tool for the quality control of Chinese traditional medicine containing glibenclamide.

Qingkailing Oral Tonic and Shuanghuanglian Oral Tonic are two kinds of complex formlae of Chinese traditional patent medicine, and both contain Scutellaria baicalensis Georgi. Baicalin is referred to the main active component with functions of dispelling wind-heat to reduce fever and detoxicating, and takes effect on uppering respiratory tract infection and tonsillities. In this work a high performance capillary electrophoresis (HPCE) method was established for separation and quantitative analysis of baicalin in prescriptions containing Scutellaria baicalensis Georgi, based on the mode of capillary zone electrophoresis. Electrophoretic conditions are as follows: uncoated fused silica capillary (34.8/39.5 cm x 50 microns i.d.), 40 mmol/L borax buffer(pH 9.0), applied voltage 17 kV, polarity anode (inlet) to cathode (outlet), temperature 25 degrees C, detection wavelength 285 nm, injection 68.95 kPa x s. Under the optimum conditions, baicalin was separated successfully from other components within 15 minutes. The relative peak area of baicalin increased linearly with the increase of its concentration in the range of 10-640 mg/L. The recoveries were (100.31 +/- 1.98)% for Qingkailing Oral Tonic and (100.60 +/- 2.36)% for Shuanghuanglian Oral Tonic. The analytical results demonstrate the method is simple, rapid and well reproducible, and can be used as a reliable tool for the quality control of Chinese traditional medicines containing Scutellaria baicalensis Georgi.

Yinhuang Granule is a kind of complex formulae made up of extract of Flos lonicerae and that of Radix scutellariae, of the fuctions of dispelling wind-heat to reduce fever, detoxicating, relieving inflammation, promoting urination and tranquilizimg the mind and can be applied to upper respiratory tract infection and tonsillitis. Baicalin and chlorogenic acid are referred to main effective components of it. In this paper, the separation and determination of baicalin and chlorogenic acid in Yinhuang Granule by capillary electrophoresis is described. The experimental conditions are stated as follow: separation column: bare fused-sillica capillary (50 microns i.d., 370 microns o.d., total length 47 cm, 40 cm from the inlet to UV detector); electrolyte: 25 mmol/L borax, pH 8.5; detection wavelength: 310 nm; sampling: 17 kPa.s; running voltage: 25 kV, polarity from anode (inlet) to cathode (outlelt); temperature: 25 degrees C; internal standard: p-nitrobenzoic acid. The linear ranges of determination for baicalin and chlorogenic acid were 160-960 mg/L(r = 0.9993, RSD = 1.76%-2.33%) and 80-960 mg/L(r = 0.9989, RSD = 1.07%-2.51%), respectively. The recoveries of baicalin and chlorogenic acid were (102.09 + 1.74)% (RSD = 1.71%, n = 6) and (99.81 + 3.11)% (RSD = 3.12%, n = 6). The method is simple, fast and accurate. It can be used for the quality control of Yinhuang Granule and its related Chinese medicines.

The derivatives of antipyrine and phenylbutazone are important antipyretic analgesics commonly used in clinical medicine. Although high performance liquid chromatography has been the conventional method used for the analysis of these drugs, in recent years capillary electrophoresis was validated to be a useful method in the analysis of antipyretic analgesics. However, there has been no report on the separation of antipyrine (AP), 4-aminoantipyrine (4-AAP), aminopyrine (APY), dipyrone (DIP) and phenylbutazone (PHE) in the literature. In this paper, a micellar electrokinetic capillary chromatographic (MECC) separation method was described for the five antipyretic analgesics.