Three butylphthalide derivatives were isolated from the Rhizome of Ligusticum chuanxiong using a series of isolation and purification approaches including macroporous resin, ODS-A column, Sephadex LH-20 and preparative HPLC. These structures were elucidated based on extensive spectroscopic data (UV, IR, HR-ESI-MS and NMR) and identified as (3Z,3aE)-(6R,7R,2'S)-6-hydroxy-7-(2-carboxyl-2-hydroxyethylthio)-3-(2-hydroxybutylidene)-4,5,6,7-tetrahydro-phthalide (1), (3Z,3aZ)-3-butylidene-6,7-dihydroxy-4,5,6,7-tetrahydro-phthalide 7-O-α-D-glucopyranosyl-(1→2)-β-D-fructo-furanoside (2) and 3-(3-β-D-glucopyranosyloxy-butylidene)-7-hydroxy-phthalide (3).

Objective: To optimize preparation techniques for Herba E pimedii flavonoid phytosomes (EFP) and explore their suitable pharmaceutics. Methods: To optimize the preparation conditions by means of unifo rm design and step regression, prepare Herba Epimedii total flavonoid phytosomes by means of solvent evaporation and investigate the accumulative dissolution of different ratios of EFP-PVP precipitates by means of dissolution release. Resu lt: The optimized preparation conditions are as follows: solvent-tetrahydrofura n, lecithin to PVP-2.5 times, temperature-40　℃ and reaction-3 hours. Oil/wa ter apparent partition coefficient of icariin was enhanced more than 4 times by phospholipid. The accumulative dissolution of Herba Epimedii flavonoids of EFP- PVP precipitate was significantly higher than that of its physical mixture and H erba Epimedii extract tablet. Conclusion: Phospholipid can ef fectively enhance the oil/water apparent partition coefficient of icariin, and P VP can improve the dissolution of Herba Epimedii phytocomes, but the pharmacokin etics needs further study.

OBJECTIVETo study the chemical constituents from root of Prismatomeris tetrandra.

METHODCompounds were isolated by chromatographic techniques on silica gel column. Their structures were elucidated by chemical and spectral methods.

RESULTSix compounds were identified as 1-hydroxy-2-methylanthraquinone (I), 2-hydoxy-3-methoxyanthraquinone (II), 1, 3-dihydroxy-2-methoxyanthraquione (II), rubiadin (IV), rubiadin-1-methyl ether (V) and beta-sitosterol (V).

CONCLUSIONI, II and III were isolated from the plant for the first time.

Anthraquinones ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rubiaceae ; chemistry

OBJECTIVETo study the chemical constituents from the roots of Cudrania cochinchinensis.

METHODThe chemical constituents of C. cochinchinensis were isolated and purified by silica gel, polyamide column chromatography, Their structures were identified on the basis of spectroscopic data.

RESULTSix compounds were isolated and identified. Their structures were established as 3, 5, 7, 4"-tetrahydroxyflavanone-7-O-(6"-acetyl)-glucoside (1), 3, 5, 7, 4'-tetra hydroxyflavanone-7-O-glucoside (2), 2', 4', 5, 7-tetrahydroxy-6-prenyldihydroflavanone (3), 5, 7, 4'-trihysdroxy-6-prenylisoflavanone (4), 1, 3, 5, 6-tetrahydroxyxanthone (5), stilbene-2, 4, 3', 5'-tetraol (6).

CONCLUSIONCompound 1 was isolated from this genus, while compounds 4 and 5 were isolated from this plant for the first time.

Flavanones ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Moraceae ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Xanthones ; chemistry ; isolation & purification

OBJECTIVETo investigate the regulatory mechanism of the transcription of tumor metastasis suppressor gene TMSG-1.

METHODSLuciferase reporter assay and site-directed mutagenesis were used to analyze the regulatory region of TMSG-1. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were carried out to verify the interaction of KLF6 and Sp1 with the regulatory region of TMSG-1. Co-immunoprecipitation (CoIP) was performed to analyze the interaction between KLF6 and Sp1. TMSG-1 and wt-KLF6 mRNA expressions in cells with different metastatic capacities were quantitated by real-time PCR. Cell invasive capability was determined by Matrigel invasion assay.

RESULTSA 63 bp inducible regulatory region (+59 bp - +123 bp) in exon 1 was identified by luciferase assay using reporter plasmids with a series of TMSG-1 regulatory region deletions. Mutations in KLF6/Sp1 binding sites of this region resulted in a decrease of luciferase activity, while cotransfection with KLF6 or Sp1 expressing plasmids led to a remarkable increase of luciferase activity. EMSA and ChIP demonstrated that KLF6 as well as Sp1 interacted with this region. CoIP also indicated a possible interaction between KLF6 and Sp1 proteins. In the highly metastatic cell sublines, a low level of wild type KLF6 was associated synchronously with a low TMSG-1 level. Prostate carcinoma cells overexpressing KLF6 exhibited a higher TMSG-1 level and a lower invasive capability.

CONCLUSIONSTranscription factor complex of KLF6 and Sp1 may participate in the inducible transcriptional regulation of TMSG-1, and a decreased wild type KLF6 expression is likely associated with a low TMSG-1 level in the highly metastatic cell sublines.

Binding Sites ; genetics ; Cell Line, Tumor ; Electrophoretic Mobility Shift Assay ; Humans ; Immunoprecipitation ; Kruppel-Like Factor 6 ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mutagenesis, Site-Directed ; Mutation ; Neoplasm Invasiveness ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Sp1 Transcription Factor ; genetics ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transcriptional Activation ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo study the survival and growth of Vibrio cholerae inside the Acanthamoeba polyphage.

METHODSSurvival and growth of Vibro cholerae O139, co-cultured with Acanthamoeba polyphaga, was observed inside the trophozoites and cysts, using Gram stain and electron microscope.

RESULTSViable O139 was observed inside the amoebal vacuoles in 24 hours. Vacuoles were filled with more bacteria along with the longer period of co-culture. The process of O139 infection with Amoebae would include uptake, formation of O139 vacuole, multiplication, trophozoites lysed and expel under electron microscopy. Some infected trophozoites could subsequently encyst and the surviving O139 could locate in the vesicles inside the cysts.

CONCLUSIONO139 might survive and multiply in the trophozoites and reside inside the cysts of Amoebae, suggesting that Acanthamoebae might serve as one of the environmental hosts of Vibro cholerae.

Acanthamoeba ; growth & development ; microbiology ; ultrastructure ; Animals ; Coculture Techniques ; Colony Count, Microbial ; Culture Media ; Vibrio cholerae O139 ; growth & development ; ultrastructure ; Water ; parasitology

OBJECTIVETo explore the role of preoperative blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) and diffusion tensor tractography (DTT) to identify the relationship between motor cortical area, pyramidal tracts with brain gliomas in neurosurgical treatment of intraoperative electrical stimulation for gliomas involving motor areas at 3T.

METHODSTwenty-eight patients with brain gliomas involving motor areas were included. They underwent MRI examination, which included conventional T1WI, T2WI, BOLD-fMRI of bilateral hands movement paradigm and diffusion tensor imaging (DTI). The data of BOLD-fMRI and DTI were transferred to the workstation (Leonardo syngo 2003A, Siemens) and analyzed. Activation mapping of hands movement, fractional Anisotropy (FA) Color and three dimensional pyramidal tracts were produced. The relationship between motor cortical area, pyramidal tracts and brain gliomas was demonstrated, which was used to optimize the pre-surgical planning. With guidance of the result of BOLD-fMRI and DTT, all patients received microsurgery under anaesthesia retaining consciousness using intraoperative motor functional brain mapping with the method of direct electrical stimulations. The brain lesions were removed as far as possible in the case of eloquent areas and sub-cortical important white matters preservation. The preoperative and postoperative KPS of all patients were operated to evaluate the state of patients.

RESULTSBOLD-fMRI, DTI were performed successfully in 28 patients. The relationship between the primary motor cortex, premotor area, supplementary motor area, pyramidal tracts and brain gliomas localized by preoperative fMRI and DTI. Under anaesthesia retaining consciousness, the primary motor area was monitored by the method of direct electrical stimulations with the guidance of preoperative BOLD-fMRI. There was good correlation between preoperative fMRI and intraoperative cortical stimulation. Furthermore, the preoperative mappings and DTT could make up for the un-monitored motor areas and pyramidal tracts during operative cortical stimulation. Comparing the preoperative KPS, the postoperative KPS was advanced.

CONCLUSIONSBOLD-fMRI and DTT could non-invasively localize the relationship between brain motor cortex, pyramidal tracts and brain gliomas in vivo to optimize the surgical planning, guide the microsurgery under anaesthesia retaining consciousness using intraoperative motor functional brain mapping with the method of direct electrical stimulations and remove brain tumors as far as possible in the case of eloquent areas and sub-cortical important white matters preservation.

Adult ; Brain Neoplasms ; pathology ; physiopathology ; surgery ; Diffusion Magnetic Resonance Imaging ; methods ; Echo-Planar Imaging ; methods ; Female ; Glioma ; pathology ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; Neuronavigation ; methods ; Transcutaneous Electric Nerve Stimulation ; methods

OBJECTIVETo study the effect of PDGF on dermal blood vessel reconstruction by transplanted tissue-engineering skin containing PDGF-B gene to rats.

METHODSThe recombined eukaryotic expression vector, pcDNA3.1-hPDGF-B, was constructed and transfected into fibroblasts mediated by LipofectAMINE. Keratinocytes + acellular dermal matrix (group A), keratinocytes + acellular dermal matrix + fibroblasts (group B), keratinocytes + acellular dermal matrix + fibroblasts with PDGF gene (group C) were recombined respectively, then transplanted them to rat dorsum and evaluated the reconstruction of blood vessels in the dermis after 2, 4, 6 week postoperation.

RESULTSIn 2-4 weeks after skin grafting the vascularization rate in group C was higher than that of group B and group A. The vascularization rates in all groups had no significant differences in six weeks (P > 0.05).

CONCLUSIONPDGF-B gene plays an important role in reconstruction of blood vessels in the dermis at early tissue-engineering skin grafting, which ensures the take of grafted tissue-engineering skin.

Acellular Dermis ; Animals ; Male ; Neovascularization, Physiologic ; Proto-Oncogene Proteins c-sis ; genetics ; Rats ; Rats, Sprague-Dawley ; Skin ; blood supply ; Skin Transplantation ; Skin, Artificial ; Swine ; Tissue Engineering ; methods ; Transfection

OBJECTIVETo observe the change of erythrocyte theology in rabbits with acute renal failure (ARF).

METHODSThirty-eight healthy rabbits were randomly divided into control group (n = 8), model group (establishing ARF model via intramuscular injection 1% HgCl2, and divided into 12 h, 24 h, 48 h subgroups, all n = 10), the arterial blood sample was taken out through carotid artery at corresponding times after anesthetization with urethane, for detecting the indices of renal function and erythrocyte rheology.

RESULTSThe levels of urea and creatinine in plasma of model rabbits at 12 h, 24 h and 48 h were higher than those of control group, and there was a rise trend along with the time extension. The erythrocytes electrophoresis time at 12 h of model group was higher, the electrophoresis rate and migration rate of erythrocytes were lower compared with those of control group; the erythrocytes electrophoresis time at 24 h of model group was lower and the electrophoresis rate and migration rate were higher compared with those of model group at 12 h; and there were no statistical differences in erythrocytes electrophoresis indices between model group at 48 h and other groups. Meanwhile, there was a rise trend in erythrocyte sedimentation rate (ESR), K value of equation and emendation along with the time extension of ARF, but these indices only at 48 h of model group was lower significantly than that of control group. There were no statistical differences in aggregation index and deformability index of erythrocytes among groups.

CONCLUSIONDuring the process of ARF, the erythrocytes electrophoresis time lengthen and electrophoresis rate and migration rate decrease at early stage, and these indices gradually return to normal; the indices of ESR increase gradually.

Acute Kidney Injury ; blood ; physiopathology ; Animals ; Erythrocyte Indices ; Erythrocytes ; physiology ; Hemorheology ; Rabbits

Objective To understand the health status and potential impact resulted in the second stage of Three Gorges Reservoir Areas sluicing. Methods Data were collected on deaths, prevalence rates of infectious and endemic diseases, as well as on vector surveillance through the project entitled 'Three Gorges Population Health Survey System'. Results The main causes of death in the population living in the Three Gorges Reservoir Areas would include: circulatory system diseases, tumors, respiratory system diseases, injuries/poison and digestive system diseases. The number of deaths caused by the above said five kind of diseases accounted for 90.94% of the total number of deaths. The prevalence rates on Water-born diseases related to the sluicing of reservoir and zoonosis-bome diseases related to the changes of vectors were still low. The indoor and outdoor densities of rodents were 3.11% and 3.16%, both were higher than that in 2006 but lower than the average numbers in the five years prior to the sluicing. The constituent ratio of Apodemus agrarius had constantly risen since 2006. The density of mosquitoes found in livestock barns and human households was higher than that in 2006 but lower than the average number of the five-year studies prior to the sluicing. Conclusion Environment change after the sluicing of the Three Gorges Reservoir Areas did not seem to have obvious impact on the health status of the people living in the areas. However, to strengthen the surveillance on the biological features of the vectors which might have related to the transmission of diseases would be highly recommended.