Aim To develop an in vitro high throughput drug screening system based on reporter gene assay for identification of novel compounds with PXR, FXR and LXRα agonist activity. Methods The expressions of exogenous PXR, FXR and LXRαgene in HEK293, HepG2 and LS174T cells were examined by Real-Time quantity PCR. pSG5-hPXR and pGL3-XREM-CYP3A4, pEGFP-N3-hFXR and EcRE-TK-Luc, pCMX-FLAG-hLXRα and pGL3-XREM-CYP3A4 were cotransfected into cells and the optimal ratio of three plasmids was determined. The dose-response relationship between the positive drug and the fold induction was determined. The specificity of the model was ex-amined, and the repeatability was also determined by Z′ value. Results ① The PXR, FXR and LXRα mRNA expression in HEK293 cell is low among three different cells. ②reporter gene vector and expression plasmid ratio of 1∶ 1, 2∶ 1 and 2∶ 1 were proved to be suitable for highest relative luciferase activity for PXR, FXR or LXRα agonist screening model. ③ The relative luciferase activity was induced by Rif, CDCA or T0901317 in a dose-dependent manner. ④Only Rif, CDCA or T0901317 could significantly increase the relative luciferase activity in PXR,FXR or LXRα agonist screening model, no effect of other nuclear re-ceptors agonist was observed, and the values of Z′-factor for PXR, FXR and LXRαagonist screening model were 0. 58, 0. 66 and 0. 63, respectively. Conclusion An in vitro PXR, FXR and LXRα agonist high-throughput screening models are devel-oped with acceptable specificity and repeatability, and the mod-els can be used to screen PXR, FXR and LXRα agonist.

PurposeTo evaluate the significance of ultrasonic elastography strain ratio, MRI and the combination of both in diagnosis of breast tumor.Materials and MethodsFifty-four cases with single breast tumor underwent preoperative ultrasound elasticity imaging and MRI. Accuracy of ultrasound elastography strain rate ratio (SRR) of the tumor and surrounding normal breast tissue was measured by quantitative ultrasound elastography, and its combination with MRI were analyzed. ResultsThere was signiifcant differences on SRR between the benign group and the malignant group (2.24±1.28vs 4.96±1.73, t=2.648,P<0.05). Optimal threshold of ultrasonic elastography SRR in differential diagnosis of breast benign from malignant tumor was 2.41 determined by ROC curve. The accuracy of SRR, MRI and the combination of both in differentiating benign from malignant breast tumor was 81.48% (44/54), 85.19% (46/54) and 96.30%(52/54), respectively. There was no statistic difference between SRR and MRI in diagnostic accuracy (χ2=0.267,P>0.05). Combined both had higher diagnosis accuracy when compared with SR and MRI separately (χ2=6.000 and 3.967,P<0.05).Conclusion Ultrasonic elastography strain ratio is accurate and objective in differentiating benign from malignant breast tumors. It is a valuable quantitative index in clinical practice. Moreover, SRR combined with MRI can reduce the misdiagnosis rate.

Objective To compare the clinical features and results between transperitoneal laparoscopic radical prostatectomy and extraperitoneal laparoscopic radical prostatectomy.Methods Thirty-three prostate cancer patients treated with laparoscopic radical prostatectomy. Among them,21 cases had been done transperitoneally and 12 cases had been done extroperitoneally. The two different approaches were evaluated and compared in respects of operating time, estimated blood loss, complications during surgery, postoperative complications, intestinal function recovery time, catheterization time and length of hospital stay.Results All the surgeries had completed successfully without conversion to open surgery. For transperitoneal approach and extraperitoneal approach, the operating time was (299±46)min and (309±64)min, blood loss was (618±448)ml and (677±469)ml. There were 3 cases with severe blood loss, 2 cases with bladder injuries and 1 case with ureteral injury in transperitoneal approach group. There were 1 case with severe blood loss, 1 case with obturator never injury, 1 case with cysto-ureteral injury and 1 case with peritoneum injury in extraperitoneal approach group. For transperitoneal approach and extraperitoneal approach,the catheterization time was(14.6±3.8)d and (12.3±2.9)d, intestinal function recovery time was (2.7±0.7)d and (2.1±0.5)d, length of hospital stay was (17.0±3.6)d and (11.2±3.5)d, respectively.Conclusions Laparoscopic radical prostatectomy is feasible and safe in clinical practice. Extraperitoneal approach has better vision, less impact on abdominal organs, faster recovery and shorter hospital stay comparing to transperitoneal approach.

OBJECTIVE: To investigate the relationship between the ovarian response, induced by gonadotropin hormone, and levels of follicle-stimulating hormone receptor (FSHR). METHODS: The level of ovarian response was divided into three groups based on the number of follicles obtained during oocyte retrieval: poor responders (<4 follicles), normal responders (4 approximate, equals 13) and high responders(>13 follicles). FSHR mRNA was measured using reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The expression of FSHR mRNA in the poor responders was 0.54+/-0.07. This was much lower than that of the normal responders who were at 0.90+/-0.17,and the high responders who were at 1.20+/-0.45(P<0.05). CONCLUSION: Ovarian response induced by gonadotropin hormone stimulation is correlated with the level of FSHR mRNA in the granulosa cells.

OBJECTIVETo assess the diagnostic value of sperm DNA fragmentation (SDF) for male infertility.

METHODSTwo hundred and ninety-nine males attending infertility clinic were classified into 157 primary infertile cases and 142 fertile controls. Semen analysis was performed as recommended by the World Health Organization (WHO). SDF was assessed by sperm chromatin dispersion (SCD) assay, and the results were expressed as DNA fragmentation index (DFI).

RESULTSThe DFI was significantly higher in infertile males than that in fertile controls [(17.1± 9.3)% vs. (14.2± 9.0)%](P< 0.01). No significant difference was detected in the age of male and female partners, seminal volume, sperm count, motility and morphology between infertile males and fertile controls (P> 0.05). The area under the receiver operating characteristic curve (AUC) was 0.861 [95% confidence interval (CI)= 0.814-0.907] for 15.1% of SDF. The threshold level of 15.1% was derived as cut-off value to discriminate infertile men from fertile controls. By this threshold, specificity was 88.2% and sensitivity was 81.8%. The 299 men were divided into group A (n= 120) with DFI≥ 15.1% and group B (n= 179) with DFI< 15.1% based on the cut-off value. The percentage of infertile men in group A was significantly higher than that in group B (79.2% vs. 34.6%) (P< 0.01). The odds ratio (OR) for infertility in the two groups was 7.2 (95%CI= 4.2-12.3).

CONCLUSIONSperms with high-level of DNA fragmentation can impair male fertility. DFI can be used as a good diagnostic marker for male infertility.

Adolescent ; Adult ; DNA ; metabolism ; DNA Fragmentation ; Female ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Spermatozoa ; metabolism ; Young Adult

Objective To detect the feasibility of magnetically labeled swine bone marrow mesenehymal stem cells(MSCs)with SHU 555A combined with poly-L-arginine(PLL),under MR imaging in vitro and in vivo.Methods Swine mesenehymal stem cells were isolated and culture-expanded 3 passages in vitro,then magnetically labeled by incubation with SHU 555A(25?g Fe/ml,Resovist,Schering)for 24 hours with 750 ng/mL poly-L-lysine(PLL;average MW_275 kDa)added 1 hour before incubation.Cellular iron incorporation and detention at 0 d,4 d,8 d,12 d,16 d,20 d after labeling was qualitatively assessed using Prussian blue and quantified at atomic absorption spectrometry.Cell viability was assessed by trypan-blue exclusion test.Cell suspensions underwent MR imaging with T_1-and T_2-weighted spin-echo and fast field-echo sequences on a clinical 1.5 T MR system.At last,1?10~6 SHU 555A labeled and unlabeled MSCs were transextracardially implanted into the infracted and normal myocardium approximately 2 week following the ligation of left anterior descending coronary artery in 1 swine respectively,and finally performed 1.5-T MRI within 1 week after infarction.Results①Intracytoplasmic particles stained with Prussian blue stain were detected for all cells with mean cellular iron content of(13.13?2.30)pg per cell.With division of stem cells, the stained particles decreased gradually with iron content(0.68?0.20)pg per cell.at 16 days after labeling, approximately to the prelabeled baseline values.(0.21?0.06)pg per cell(P＞0.05).The viability of the labeled cells at various time points were not significantly different with that of nonlabeled cells(P＞0.05).②MR images showed signal intensity changed most obviouly in T2*WI in vitro.The percentage change of signal intensity increased with increasing cell numbers,and decreased with the time.As few as 5?10~4-1?10~5 cells could be detected by using this approach.③Two injected sites containing MR-MSCs were detected in vivo,presentingas low signal intensity areas with the T_2*WI scanning sequence.Conclusion Swine bone marrow MSCs can be labeled with SHU555A-PLL and depicted with a standard 1.5-T MR imager in vitro and in vivo.(J lntervent Radiol,2007,16:115-121)

OBJECTIVETo evaluate the diversity of germplasm resources of Artemisia annua and provide the basis for improving utilization of germplasm resources, the agronomic traits of germplasm resources of A. annua were studied in Yun-Gui plateau.

METHODThe agronomic traits of 67 A. annua germplasm resources were measured by the visual observation and measurement methods. And the germplasm resources were clustered using flexible-beta method to analysis their genetic background.

RESULTThe result showed that 67 germplasm resources had a relatively wide variation on the 22 agronomic traits. Among 22 agronomic traits, the dry weight of branch had the greatest coefficient of variation, which was 53. 63, and the next were the dry weight of leaf, total plant weight, the length of pinnules and the length of leaflet, which were 42.74, 41.61, 39.54 and 39.22 respectively. The smallest coefficient of variation was the leaf corlor. Based the result of cluster analysis, these 67 germplasm resources were classed into 5 groups, and each group had its respective character. The first group showed early-maturing resources, dwarf stalk, slender rod, long bipinnata, high leaf-stem ratio and moderate leaf weight The third group showed late-maturing resources, tall and thick stalk, much-branch, bushy accessory pinna, high leaf weight and yield. The fifth group showed very late-maturing resources, strong lateral shoot, high leaf yield.

CONCLUSIONThere were significant genetic difference and diversity in the germplasm resources of A. annua. The result of cluster analysis showed that the resources of group 1, group 3 and group 5 were suitable as breeding material of A. annua.

Artemisia annua ; classification ; genetics ; growth & development ; Biodiversity ; Biomass ; China ; Cluster Analysis

Flavonoids are an important effective component of traditional Chinese medicine, which are widely distributed in the plant kingdom. The biosynthesis of flavonoid in plants is affected and regulated by various environmental factors. For a necessary environmental factor to plant growth and development, mineral nutrients are paid more and more attention on the regulation to the metabolism of flavonoids in medicinal plants. In this paper, an overview of flavonoids biosynthetic pathway, and the macroelements, microelements and rare earth elements on the metabolism of flavonoids in medicinal plants are presented. And the regulation mechanism of them are also analyzed and discussed.
Flavonoids ; analysis ; metabolism ; Minerals ; analysis ; metabolism ; Nutrition Assessment ; Plants, Medicinal ; chemistry ; metabolism

OBJECTIVETo provide the basis for improving utilization of Artemisia annua germplasm resources and breeding variety, the interrelations between artemisinin content, artemisinin yield and agronomic traits of A. annua were studied.

METHODThe artemisinin content and each agronomic trait of 63 A. annua germplasm resources were measured by the visual observation and measurement methods. And the correlation analysis, regression analysis and path analysis were adopted.

RESULTThe result showed that there were significant differences in the artemisinin content and yield of 63 germplasm resources from the main production region of A. annua. Correlation analysis showed that there were significantly positive correlation between leaf weight and artemisinin yield with stem and branch characters, but there were negative correlation between artemisinin content with leaf characters of A. annua plant. The artemisinin content of A. annua increased with the increasing of primary branch number, bottom secondary branch number, and bottom stem diameter, etc. On the other hand, it decreased with the increasing of top secondary branch number, secondary leaf axis length, and bottom branch diameter, etc. The artemisinin yield of A. annua increased with the increasing of artemisinin content, leaf weight, and bottom secondary branch number, etc., and decreased with the increasing of bottom branch diameter, middle secondary branch number, and stem weight, etc. Path analysis showed that the primary branch number and bottom secondary branch number had a direct positive effect on the artemisinin content of A. annua. But the top secondary branch number had a direct negative effect on the artemisinin content of A. annua. The leaf weight and artemisinin content had a direct positive effect on the artemisinin yield and the ratio of leaf/stem, branch weight and stem weight had a direct negative effect.

CONCLUSIONOn the breeding A. annua variety, it can take into account both high leaf yield and high artemisinin content. And it was strongly recommend that the plant with moderate plant height and crown, shortness pinnae and secondary leaf axis, less middle and top secondary branch, strong stem, higher primary branch number and bottom secondary branch number, and higher ratio leaf/stem could be selected for breeding new varieties with high leaf yield and high artemisinin content.

Artemisia annua ; chemistry ; growth & development ; Artemisinins ; analysis ; Biomass ; Plant Extracts ; analysis ; Plant Leaves ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development

OBJECTIVETo analyze the mutational features of adenomatous polyposis coli (APC) gene and to explore the effect of mismatch repair (MMR) deficiency on its mutations in hereditary non-polyposis colorectal cancers (HNPCC).

METHODSPCR-based in vitro synthesized protein test (IVSP) assay and sequencing analysis were used to confirm somatic mutations of whole APC gene in 19 HNPCC patients.

RESULTSEleven cases with thirteen mutations were determined. The frequency of APC mutation was 58%(11/19). The exhibiting mutations consisted of 9 frameshift mutations and 4 nonsense ones, indicating the existence of more frameshift mutations (69%). All of frameshift mutations were deletion or insertion of 1-2 bp and most of them (7/9) happened at simple nucleotide repeat sequences, particularly within (A) n tracts (5/9). All of four nonsense mutations resulted from C to T transitions at CpG sites.

CONCLUSIONMutational inactivations of APC gene were detected in more than half of HNPCC patients in this study, indicating that APC mutation is a common molecular event in the tumorigenesis of HNPCC. According to the location of frameshift mutations at simple nucleotide repeat sequences and point mutations at CpG sites, it was suggested that endogenous mechanisms like MMR deficiency might exert an effect on the nature of APC mutations in most HNPCC.

Adenomatous Polyposis Coli ; genetics ; Adenomatous Polyposis Coli Protein ; genetics ; metabolism ; Carcinoma ; genetics ; Colorectal Neoplasms ; genetics ; pathology ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; Genes, APC ; physiology ; Humans