Objective To investigate the role of intra-operative and percutaneous three-dimensional contrast-enhanced ultrasonic cholangiography (3D-CEUSC) in the diagnosis of biliary anomalies. Methods Diluted SonoVue, an ultrasonic contrast agent, was injected into the cystic duct of 13 living liver donors via a cannula for intra-operative 3D-CEUSC, while injection via a T tube or a PTCD tube were done in 14 patients for percutaneous 3D-CEUSC. The maximum branching order of the intra-hepatic bile ducts, the percentages of display of the first-, second-, and third-order intra-hepatic bile ducts, and the rates of detection of biliary anatomical anomalies were evaluated. Results The median of the maximum branching order of the intra-hepatic bile ducts shown was fourth-order on intraoperative 3D-CEUSC and third-order on percutaneous 3D-CEUSC, respectively. The difference was significant (P=0.01). From the first- to the third-order of intra-hepatic bile ducts, the percentages of display for intra-operative 3D-CEUSC were 100% (23/23), 96.2% (50/52) and 82.7% (86/104),respectively, while they were 100% (24/24), 94.6% (54/56) and 60. 7% (68/112) for percutaneous 3D-CEUSC. The differences were not significant when comparing the first- and the second-order branches of intra-hepatic bile ducts as shown either on intra-operative or percutaneous 3D-CEUSC (P=1).The percentages of display of the third-order branches using intra-operative 3D-CEUSC was higher than that of percutaneous 3D-CEUSC (P＜0. 0001). Three biliary anatomical anomalies were detected by intra-operative 3D-CEUSC and 4 anomalies were detected by percutaneous 3D-CEUSC. All of these anomalies were confirmed by X-ray cholangiography. Conclusion Both intra-operative and percutaneous 3D-CEUSC could diagnose biliary anatomical anomalies accurately. These procedures may become important techniques to evaluate the biliary anatomy in hepatobiliary surgery pre-operatively or intraoperatively.

efore graft harvesting in living donor liver transplantation.

Objective To investigate the feasibility of three-dimensional contrast-enhanced ultrasonic cholangiography(3D-CEUSC) in isolated porcine liver specimens.Methods The lobes of 5 isolated porcine livers were taken as scanning units and the lobular bile branches were cannulated.3D-CEUSC was performed after the administration of diluted ultrasonic contrast agents(SonoVue) through the cannula.The images were assessed in term of maximum visualization of branching orders and image quality.The coincidence of morphous was compared between 3D-CEUSC and cholangiagraphy using fluoroscopy.Results Fourteen lobular biliary branches were cannulated successfully in 5 isolated porcine livers.The maximum visualization of branching orders from the third- to first-order branches were 42.9% (6/14) ,42.9% (6/14), 14.2% (2/14) in sequence.The image quality was 21.4% (3/14) graded as well, 57.2% (8/14) as moderate, and 21.4% (3/14) as poor.Compare with cholangiagraphy using fluoroscopy, the coincidence of morphous was 42.9%(6/14) for well,42.9% (6/14) for moderate,and 14.2% (2/14) for poor.Conclusions 3D-CEUSC could be a new imaging method to evaluate the biliary anatomy.

OBJECTIVETo study the relationships among the main agronomic characters, yield per plant,content of artemisinin of Artemisia annua, and provide the theoretical basis for breeding the A. annua varieties with high-yield and high artemisinin content.

METHODThe main agronomic characters, yield per plant, artemisinin content of the wild and cultivated A. annua of Sichuan province were investigated and determined, and the correlated relationships among them were analyzed.

RESULTIt was showed that the weight of dried stem per plant had very significantly positive correlation with the yield per plant,and the number of primary branches, the diameter of stem had significantly positive correlation with the yield per plant. The flower bud stage, first flowering stage and full-bloom stage had significantly positive correlation with the artemisinin content. The first flowering stage and full-bloom stage had significantly positive correlation with the total artemisinin content per plant. The weight of dried stem per plant,the diameter of stem and the flower bud stage had significantly correlation with the total artemisinin content per plant. A. annua with the purple-stem had significant higher artemisinin content than that with the green-stem. Through the multiple regression analysis, the classic multiple regression equation of the yield per plant was gained as Y = 0.06121-0.000 08X(1) + 0.30378X(3) - 0.00051X(4) - 0.00031X5 (F = 20.15, P < 0.0001), the classic multiple regression equation of the artemisinin content as Y = -1.45849 - 0.00099X(1) - 0.24079X(3) + 0.13362X(6) - 0.00719X(7) + 0.01454X(9) (F = 46.55, P < 0.0001), the classic multiple regression equation of the total artemisinin content per plant as Y = -2.67001 - 0.00249X(1) + 0.98873X(3) + 0.30037X(6) + 0.113X(9) F = 22.62, P < 0.001).

CONCLUSIONThe plant with purple, thick and strong stem, higher plant height, moderate primary branches number and bloom stage, small stem-branch angle should be selected for a breeding new A. annua varieties with high yield and artemisinin content.

Artemisia annua ; chemistry ; growth & development ; metabolism ; Artemisinins ; analysis ; metabolism ; Biomass ; China ; Flowers ; chemistry ; growth & development ; metabolism

OBJECTIVETo provide more proofs for expounding the genetic relationships among the (varietal) species in genus Fritillaria from Sichuan province.

METHODThe ISSR marker technique was used to study relationships and genetic polymorphism of nineteen populations in ten species and one varietal species of genus Fritillaria. Genetic similarities were calculated by using NTSYS software and the dendrogram was constructed by using UPGMA method.

RESULTEleven primers were selected from 35 ISSR primers, and 179 DNA fragments were amplified from 19 populations. Of which, 179 fragments were polymorphic (percentage of polymorphic bands was 86.8%). The genetic similarity among all accessions ranged from 0.569 to 0.855. Clustering analysis showed that the 19 populations of Fritillaria could be distinctively classified into 4 groups. F. cirrhosa, F. przewalskii, F. cirrhosa var. logirnectarea and F. dajitensis were in the first group; The second group was the cluster of F. cirrhosa and F. mellea (wild and cultivated species); The third group was F. sulcisquamosa, F. thunbergii, wabunesis and F. delavayi; F. hupehensis alone formed the fourth group.

CONCLUSIONISSR marker technique is suitable for the genetic diversity of Fritillaria from Sichuan province. Interspecific identifications among the four original species of Bulhus Fritillariae Cirrhosae recorded by pharmacopoeia of China, and between them and the other species of genus Fritillaria from Sichuan province could not be gained by using ISSR markers technique. In addition, the cluster result of genus Fritillaria had some relationships with the geographical distribution.

China ; Fritillaria ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny

OBJECTIVETo study the physiologic indexes, yield and the contents of alkaloids of Fritillaria cirrhosa D. Don under different temperature.

METHODThe growth temperatures (15, 20, 25, 30 degrees C) of F. cirrhosa were controlled by using artificial climate, the growth was observed, the contents of chlorophyll a and b, soluble sugar, MAD, proline of the leaves of F. cirrhosa were tested, and the yield and the alkaloids content of the bulbs were analyzed.

RESULTThe growth period of F. cirrhosa under 15, 20 degrees C were appropriately extended. The difference of the content of leaves chlorophyll b under four temperatures and the contents of total chlorophyll and chlorophyll a under 15, 20, 30 degrees C were not significant. The contents of soluble sugar, MAD and proline of leaves and the growth ratio, dry weight and content of alkaloids of bulb increased with the temperature decrease.

CONCLUSIONHigher temperature is not suitable for the growth of F. cirrhosa. Under the relatively lower temperature, the growth period of F. cirrhosa extended, the bulb can grow properly, and the content of alkaloid increased. F. cirrhosa can improve its cold tolerance by increasing the content of proline and soluble sugar, and it also can maintain the normal content of chlorophyll under the lower temperature.

Alkaloids ; metabolism ; Fritillaria ; growth & development ; metabolism ; Gene Expression Regulation, Plant ; Temperature

Objective　To observe the changes of the microvascular permeability after blunt chest trauma (BCT), endotoxemia and their combined injury in rats. Methods　After the establishment of the rat models of BCT, endotoxemia and their combined injury in the right lungs, the fluorescein sodium (FINa) content was measured with flurospectrophotometer in lungs 0.5, 1, 4 and 8 h after injury. Results　There was an early obvious increase of the microvascular permeability in the impact lateral (peak at half an hour after injury), and a delayed increase in the contralateral lung (peak at the 8th h) in the BCT group. The FINa content was higher in endotoxemia group than in the BCT group(P<0.05), and lower than that in the combined injury group(P<0.05) in the contralateral lung. Conclusion　Results indicate that there were different pathophysiologic processes among the 3 kinds of injury and the FINa content is a useful index to manifest the changes of microvascular permeability in tissues.

OBJECTIVETo explore the therapeutic efficiency of human telomerase reverse transcriptase promoter (hTERTp) mediated horseradish peroxidase (HRP) catalyzed effects of indole-3-acetic (IAA) on laryngeal squamous cell carcinoma with different radiosensitivity in vivo.

METHODSHuman laryngeal squamous cell carcinoma Hep-2 and Hep-2R cells were transplanted into nude mice. After growing to about 30 approximately 50 mm3, the tumor-bearing mice were randomly divided into eight groups: Hep-2 line: combined group (A), gene group (B), radiation group (C) and blank group (D); Hep-2R line: combined group (AR), gene group (BR), radiation group (CR) and blank group (DR). The phTERTp-HRP was delivered by intratumoral injection and the IAA by intraperitoneal injection, combined with 2 Gy daily radiation to a total dose of 30 Gy. The tumor volume was recorded. The cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The expression of HRP protein was detected by AP immunohistochemisty.

RESULTSThe tumor growth of combined groups was attenuated significantly and the tumor volume of Hep-2R blank group was the largest. The inhibition rate of each group was: A: 54.8%, B: 10.0%, C: 31.9%; AR: 52.7%, BR: 24.8%, CR: 17.0%. In the combined groups, necrosis and apoptosis of tumor cells were observed under the light microscope and the apoptotic index [A (16.6 +/- 1.3)% vs. AR (17.6 +/- 1.3)%] of tumor cells was highest (P < 0.05). The HRP protein expression of BR (33.3 +/- 8.9)% was higher than that of B (21.9 +/- 5.7)%, which was directly up-regulated in the tumors (45.0% vs. 54.8%, P < 0.05) after radiation.

CONCLUSIONIn the Hep-2- and Hep-2R-transplantation tumors in nude mice, hTERTp can be induced by radiation and enhance the expression of horseradish peroxidase (HRP) gene according to telomerase activity. hTERTp-HRP/IAA system, which has synergistic effects with radiation and inhibits the tumor growth by induction of apoptosis and necrosis, may be a new gene-radiation strategy for the treatment of laryngeal carcinoma.

Animals ; Apoptosis ; Cell Line, Tumor ; Combined Modality Therapy ; Female ; Genetic Therapy ; methods ; Horseradish Peroxidase ; genetics ; metabolism ; Humans ; Indoleacetic Acids ; metabolism ; pharmacology ; Laryngeal Neoplasms ; metabolism ; pathology ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Promoter Regions, Genetic ; Radiation Tolerance ; Radiotherapy ; methods ; Random Allocation ; Telomerase ; genetics ; Tumor Burden

Critical Illness ; Extracorporeal Membrane Oxygenation ; Hemodynamics ; Humans ; Infant, Newborn ; Male

OBJECTIVETo explore the synergistic anti-tumor effect of radiotherapy and horseradish peroxidase/prodrug indole-3-acetic acid (HRP/IAA) gene therapy system using chimeric hTERT promoter responsive to ionizing radiation.

METHODSThe synthetic hTERT promoters containing four tandem-repeat copies of radio-inducible CArG elements, and the chimeric promoter containing cytomegalovirus (CMV) early promoter were both constructed. The activities of the chimeric promoters in cancer cell lines (HeLa, A549, and MHCC97) and normal cell line (MRC-5) were detected using luciferase reporter gene expression analysis after a (60)Co γ-irradiation treatment at a series of doses(a single dose of 0 to 10 Gy). The anti-tumor effect of combining irradiation with HRP/IAA gene-directed enzyme prodrug therapy system controlled by the chimeric promoter was tested by colony formation assay, cell counting and apoptosis analysis.

RESULTSThe chimeric promoters were ineffective in normal human cells, even after irradiation, but the expression of luciferase gene in tumor cells was significantly higher. The activity of the chimeric promoter in MRC-5 cells was 22.3%, 12.9% and 13.6% of that in HeLa, A549 and MHCC97 cells, respectively. After irradiation, the ratios were 11.7%, 8.7% and 8.8%, respectively. Furthermore, the chimeric promoters could successfully induce the expression of luciferase gene following different doses of radiation, with maximal inducible activity seen after 6 Gy irradiation. The chimeric promoter containing four tandem-repeat copies of radio-inducible CArG elements and CMV early promoter showed the highest activity with 6 Gy irradiation. The relative luciferase activities in HeLa, A549 and MHCC97 cells were 1.7 ± 0.2, 2.3 ± 0.2 and 2.3 ± 0.1, respectively. The chimeric promoter mediated suicide gene therapy system could increase radio-sensitivity in different cancer cells. Compared with the control system, it plus irradiation showed stronger cell proliferation inhibition, 67.3% vs. 26.1% in HeLa, 69.0% vs. 28.3% in A549, 64.6% vs. 20.8% in MHCC97 cells, and also higher apoptosis-inducing effect, 39.6% vs. 14.2% in HeLa, 33.0% vs. 12.4% in A549, and 33.2% vs. 14.2% in MHCC97 cells.

CONCLUSIONSChimeric promoter containing hTERT promoter, CArG elements and CMV promoter preserve the tumor-specificity in telomerase-positive tumor cells, and irradiation-responsive to low dose of radiation. The suicide gene therapy using this promoter plus radiotherapy show a strong anti-tumor effect in vitro. It is expected to have a good potential for future application in gene radiotherapy.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Combined Modality Therapy ; Cytomegalovirus ; genetics ; Genes, Transgenic, Suicide ; Genetic Therapy ; methods ; Genetic Vectors ; Horseradish Peroxidase ; genetics ; metabolism ; Humans ; Indoleacetic Acids ; metabolism ; Luciferases ; genetics ; metabolism ; Plasmids ; Prodrugs ; Promoter Regions, Genetic ; radiation effects ; Radiotherapy ; methods ; Recombinant Proteins ; genetics ; metabolism ; Telomerase ; genetics ; metabolism ; Transfection