Virginiamycin acetyltransferase D (VatD) plays a vital rule in streptogramins resistance by chemically inactivating streptogramin A. Therefore, it is desirable to discover novel small molecular weight inhibitors of VatD via state-of-the-art virtual screening techniques. This "cocktail" strategy by combining VatD inhibitor with streptogramins may provide new therapeutic opportunity for resistant bacteria infections. Structure-based virtual screening method (molecular docking) was applied to rank and score a chemical database containing 300 000 commercially available compounds against the VatD substrate binding site. Twenty six out of the 200 top scored compounds from the docking calculation were selected and submitted to the VatD enzymatic inhibition assay. The plasmid pRSET B/vatD was constructed and transformed into E. coli (trxB) host cells for over-expression, and VatD enzyme was purified and validated by showing acetyltransferase activity to Virginiamycin M1. Three out of these 26 tested compounds showed enzymatic inhibition on VatD with IC50 168.6, 91.0 and 55.2 micromol x L(-1), separately. Other compounds could not be dissolved in the system and/or had little effect on the enzyme (IC50 > 200 micromol x L(-1)). To our knowledge, it is first time that small molecular weight organic compounds were identified as VatD inhibitors. It is expected that the VatD inhibitors identified at present study could serve as lead compounds for the further development of the novel therapeutic agents to overcome streptogramins resistance.
Acetyltransferases ; antagonists & inhibitors ; genetics ; metabolism ; Catalysis ; drug effects ; Drug Design ; Drug Resistance, Bacterial ; Enzyme Inhibitors ; chemistry ; metabolism ; pharmacology ; Escherichia coli ; genetics ; Genetic Vectors ; Kinetics ; Molecular Structure ; Plasmids ; Streptogramin Group A ; chemistry ; metabolism ; pharmacology ; Transformation, Genetic

This paper aimed to study the effect nitrogen supplying on biomass accumulation and root respiration dynamic change of Glycyrrhiza uralensis and reveal the metabolic pathway of root respiration impact the biomass accumulating of G. uralensis. Six groups of one-year-old G. uralensis were fertilized with total nutrition containing various nitrogen concentration (0, 0.5, 1, 2, 4, 8 mmol x L(-1)) every week. At the end of every month, from June to October, the volume respiration rate and biomass of different classes of root samples were determined, and the correlation between root respiration and biomass was analyzed. The results indicated a negative correlation between volume respiration rate and biomass, nitrogen supply significantly affected both root respiration and biomass of G. uralensis by reducing root respiration and increasing root biomass. Under 8 mmol x L(-1) nitrogen supplying, there existed the optimal inhibition of root respiration, which has increased biomass of G. uralensis.
Biomass ; Dose-Response Relationship, Drug ; Glycyrrhiza uralensis ; drug effects ; growth & development ; metabolism ; Kinetics ; Nitrogen ; pharmacology ; Oxygen Consumption ; drug effects ; Plant Roots ; drug effects ; metabolism ; Seasons ; Time Factors

A new compound and five known compounds were isolated from the ethanolic extract of the leaves of Ilex pernyi Franch. Their structures were established on the basis of spectral analysis and identified as trans-isoeugenyl-alpha-L-arabinopynosyl (1 --> 6) -beta-D-glucopyranoside (1) , kaempferol-3-O-sambubioside (2), quercetin-3-O-sambubioside (3), isoquercitrin (4), (+) -syringaresinol-O-beta-D-glucopyranoside (5), amarantholidoside IV (6). Among them, compound 1 is a new phenolic glycoside, named as ilexperphenoside A, and compounds 2-6 were isolated from this plant for the first time.
Glucosides ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Ilex ; chemistry ; Molecular Structure ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analogs & derivatives ; chemistry ; isolation & purification

The purpose of this study is to investigate the activity and mechanism of a new anti-tumor agent T03. MTT and colony formation assay were performed to determine anti-proliferation activity of T03 in vitro. Antitumor activity was observed by Renca xenograft model in vivo. The effect of T03 on cell cycle and apoptosis were measured by FCM analysis. Western blotting was performed to investigate the expression level of proteins in HepG2 cell lines treated with T03. T03 had anti-tumor activity by inhibiting tumor cell growth and colony formation in vitro, especially on hepatocellular carcinoma cells (HCC). At the concentration of 10 micromol x L(-1), T03 induced cell apoptosis and cell cycle arrest in HCC. Moreover, it proved that T03 reduced the tumor weight with the rate of 42.30% without any obviously side effect in Renca xenograft model. At the concentration of 2.0 micromol x L(-1), T03 was able to reduce the level of p-c-Raf (Ser259), and thus blocked Raf/MEK/ERK and AKT signaling in HepG2 cell lines. The result suggested that T03 has the potential to inhibit cell proliferation and induce cell apoptosis both in vitro and in vivo, particularly active against HCC, indicating T03 and its analogues may serve as a new anti-cancer drug against hepatocellular carcinoma.
Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Hep G2 Cells ; drug effects ; Humans ; Liver Neoplasms ; pathology ; Signal Transduction ; drug effects ; Xenograft Model Antitumor Assays

Objective To assess the value of real-time intraoperative neuronavigation integrated with ultrasound in the resection of deep-seated brain tumor. Methods Thirty patients with deep-seated brain tumor were treated with microneurosurgery guided with real-time ultrasound integrated with the Brain Lab IGSonic navigation. During the ultrasound based operation, the degree of brain shift and the tumor border was timely observed, and then the tumor was resected totally. Results Guiding with navigation integrated ultrasound, we noticed the brain shift with various degrees happening in 30 patients,and the border of tumor was exposed and the tumor was resected totally without serious complications.Conclusions Intraoperative ultrasound integrated with navigation is a reliable guidance which can accurately re-localize the border of deep-seated brain tumor even when the tumor is shifting, timely delineate the reformatted images from ultrasound and totally resectcd the tumor, thus decrease the surgical time and increase the safety of surgical procedure.

OBJECTIVETo investigate the associations of polymorphisms of metabolic enzyme genes with urinary 1-hydroxypyrene levels in coke oven workers.

METHODSOne hundred and forty-eight workers from a coke oven plant and 69 controls without occupational PAHs exposure were selected in this study. Urinary 1-hydroxypyrene was detected by high performance liquid chromatography with florescence detector. The genotypes at I462V site in exon 7 of CYP1A1 gene, GSTM1, GSTT1, I105V site in GSTP1gene, Pst1 and Dra1 sites in CYP2E1 gene, P187S site in NQO1 gene, Kpn1, BamH1 and Taq1 sites in NAT2 gene, and H113Y, R139H sites in mEH gene were determined by PCR-based methods. Personal information including occupational exposure history, age, sex, smoking and drinking status was collected by the questionnaire.

RESULTSThe level of urinary 1-hydroxypyrene in coke oven workers [(5.61 +/- 1.04) mol/mol Cr] was higher than that in control [(0.74 +/- 0.32) micro mol/mol Cr]. After adjusting external occupational exposure category and smoking, coke oven workers with variant homozygotes at H113Y site of mEH gene had significantly higher urinary 1-hydroxypyrene concentrations than those with heterozygotes, and wild homozygotes (6.41 +/- 1.09 vs. 6.24 +/- 1.08, and 4.62 +/- 0.95 micro mol/mol Cr, P < 0.05), and gene-gene interaction was found between CYP1A1 and mEH.

CONCLUSIONGenetic polymorphism of mEH gene could be a susceptible biomarker in coke oven workers which was involved in the individual susceptibility on metabolism of PAHs.

Coke ; adverse effects ; Cytochrome P-450 CYP1A1 ; genetics ; DNA Damage ; genetics ; Epoxide Hydrolases ; genetics ; Genetic Predisposition to Disease ; genetics ; Glutathione Transferase ; genetics ; Humans ; Male ; Occupational Exposure ; Polycyclic Aromatic Hydrocarbons ; poisoning ; Polymorphism, Genetic ; Pyrenes ; analysis ; metabolism

OBJECTIVETo investigate the association of polymorphisms of metabolic and DNA repair enzyme genes and DNA damage in peripheral blood lymphocytes in coke-oven workers.

METHODSOne hundred and forty-four coke-oven workers and 50 controls were recruited in this study. Urinary 1-hydroxypyrene (1-OHP) levels were measured as the internal dose of polycyclic aromatic hydrocarbons exposure. DNA damage was detected by alkaline comet assay, and the value of 1.74 was used as the cut-off value to determine whether the individual's DNA damage was positive. The genotypes of CYP1A1, CYP2E1, GSTP1, NQO1, mEH and XRCC1 were determined by PCR-based methods. With adjustment for urinary 1-OHP, age, sex, multiple analysis of covariance was used to study the association between genotypes and the ln-transformed olive TM and multiple logistic regression was used to calculate the adjusted OR and the 95% CI for the risk of DNA damage.

RESULTSIn 144 coke-oven workers, with adjustment for urinary 1-OHP, coking history and sex, the olive TM was significantly higher with XRCC1 280His allele than those with Arg allele (5.6 vs. 2.8, P < 0.01). The subjects with XRCC1 280His allele also have significantly higher risk for DNA damage than subjects with Arg allele (adjusted OR = 2.66, 95% CI = 1.00-7.14, P = 0.05) and the subjects with GSTP1 104Val allele have higher risk for DNA damage than subjects with Ile allele (adjusted OR = 1.90, 95% CI = 0.94-3.85, P = 0.07).

CONCLUSIONXRCC1 and GSTP1 polymorphisms might influence the susceptibility of DNA damage in occupational PAH-exposed coke-oven workers.

Adult ; Case-Control Studies ; Coke ; poisoning ; Comet Assay ; Cytochrome P-450 CYP1A1 ; genetics ; Cytochrome P-450 CYP2E1 ; genetics ; DNA Damage ; DNA Ligase ATP ; DNA Ligases ; genetics ; DNA Repair Enzymes ; genetics ; Female ; Genotype ; Glutathione S-Transferase pi ; genetics ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; NAD(P)H Dehydrogenase (Quinone) ; genetics ; Occupational Exposure ; adverse effects ; analysis ; Polymorphism, Genetic

OBJECTIVETo observe effects of removing arms and ovarian on lumbar intervertebral disc and vertebral bone mineral density (BMD) by establishing rat model of lumbar intervetebral disc degeneration (IDD) with kidney deficiency, and to explore internal mechanism of disc degeneration, relationship between disc degeneration and osteoporosis.

METHODSThirty Sprague-Dawley female rats aged one month were randomly divided into control group, lumbar IDD group and lumbar IDD with kidney deficiency group (combined group), 10 rats in each group. Lumbar IDD group removed double arms, lumbar IDD with kidney deficiency group removed double arms after 3 months, both ovaries were removed. Vertebral bone mineral density were observed by Micro-CT scan; morphological changes were tested by safranine O-fast green staining; II, X collagen protein expression in the intervertebral disc were obsevered by immunohistochemistry; extracellular matrix gene expression were obsevered by real-time polymerase chain reaction (RT-PCR), in order to evaluate the effects of removed of forelimbs and double ovarian on degeneration and vertebral bone mineral density of intervertebral disc.

RESULTSMicro-CT scan showed osteoporosis in kidney deficiency group was obviously worse than other two groups; safranine O-fast green staining showed that intervertebral space became narrowed, intervertebral disc tissue degenerated obviously, chondral palte was underdeveloped in kidney deficiency group; immunohistochemistry showed that X collagen expression increased, type II collagen expression decreased in kidney deficiency group; RT-PCR showed that type II collagen expression in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P=0.000); Age 1 in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P= 0.000); while type X collagen expression was higher than control group, but no significant meaning; MMP-13 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P= 0.000, P=0.000); aggrecanase-2 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P=0.006, P=0.008).

CONCLUSIONRats model of lumbar disc degeneration established by removed forelimbs and ovariectomized can occure "bone like"--osteoporosis, which is similar with clinical kidney lumbar disc degeneration in tissue morphology, molecular cell biology expression.

Animals ; Collagen ; genetics ; metabolism ; Extracellular Matrix ; genetics ; metabolism ; Female ; Humans ; Intervertebral Disc Degeneration ; etiology ; metabolism ; physiopathology ; surgery ; Kidney ; physiopathology ; Osteoporosis ; complications ; genetics ; metabolism ; Ovariectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the association of gene polymorphism of organophosphate insecticides (OPs) metabolic enzymes with intermediate myasthenia syndrome (IMS) following acute OPs poisoning.

METHODSThirty six of 147 acute OPs poisoning patients developed IMS one to four days after poisoning. Peripheral blood samples were collected from all the patients and whole blood cholinesterase (ChE) activity was determined by DTNB spectrometry. The genetic polymorphism of CYP2E1 (1091C-->T) and GSTP1 (313A-->G) were analyzed by polymerase chain reaction (PCR)-restrict fragment length polymorphism, CYP1A1 (4889A-->G), GSTM1 and GSTT1 by allele-specific PCR, and PON1 at 55 codon (55L-->M) by PCR-single strand conformation polymorphism.

RESULTSThe whole blood ChE activity in IMS patients was not significantly different from non-IMS patients at admission (38.22 +/- 17.56)% and (42.49 +/- 16.23)%, respectively, P > 0.05, but recovered much slower in IMS patients than that in non-IMS patients. The frequencies of heterozygote and variant homozygote of PON1 at 55 codon, GSTM1 null, and both GSTM1 and GSTT1 null were higher in IMS patients than those in non-IMS patients (P < 0.05), with odds ratios and their 95% confident intervals of 2.48 (1.06 - 5.78), 11.23 (2.95- 42.76), 2.53 (1.14 - 5.61) and 2.68 (1.20 - 5.97), respectively.

CONCLUSIONSPatients of OPs and its mixture poisoning with genotype of variant allele at 55 codon of PON1, GSTM1 null and both GSTM1 and GSTT1 null probably had higher risk for IMS.

Adult ; Cholinesterases ; metabolism ; Cytochrome P-450 CYP2E1 ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Glutathione Transferase ; genetics ; Homozygote ; Humans ; Insecticides ; poisoning ; Male ; Middle Aged ; Myasthenia Gravis ; chemically induced ; genetics ; Organophosphorus Compounds ; Point Mutation ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Syndrome

OBJECTIVETo observe the relationship between protein sythesis and cardiomyocyte viability in neonatal rats.

METHODSThe protein sythesis in neonatal rat cardiomyocytes was measured according to Brandford's method, the absorbance at 490 nm (A(490 nm)) of the cells was measured with MTT assay and the cell viability evaluated by the ratio of A(490 nm) to the total cell number.

RESULTSET-1 increased cardiomyocyte protein synthesis dose-dependently, and this effect was attenuated by the application of lacidipine and tetramethylpyrazines Higher doses of ET-1 resulted in lower A(490 nm)/total cell number ratio, which was further lowered by larcidipine and tetramethylpyrazine.

CONCLUSIONThe status of protein synthesis is not associated with the viability of neonatal rat cardiomyocytes.

Animals ; Animals, Newborn ; Calcium Channel Blockers ; pharmacology ; Cell Survival ; drug effects ; Cells, Cultured ; Dihydropyridines ; pharmacology ; Dose-Response Relationship, Drug ; Endothelin-1 ; pharmacology ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Protein Biosynthesis ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley