To establish a HepG2 cell line ,which stably expressing CYP3A29 of Bama miniature pig in order to evaluate the drug metabolic characteristics of this isoenzyme,the gene for this system was obtained through total RNA extraction and RT-PCR assay.the gene was subcloned into plasmid PMD18-T,designated as pMD-CYP3A29.This gene was then amplified by PCR, and cloned into eukaryotic expression vector pcDNA3.1(+),and the recombinant plasmid was designated as pcDNA-CYP3A29. Sequencing was used to confirmed the correctness of the gene of this gene.the expressed gene was then transfected into HepG2 cells by lipid-media transfection and the transformants were screened by G418 for 10 generations ;the expression of CYP3A29 was identified by RT-PCR、West-blot and the metabolic activity of the transformant HepG2-CYP3A29 was verified by nifedipine oxidation.In comparison with HepG2, the transformant HepG2-CYP3A29 showed remarkable oxidative activity.It is apparent that the cell line stably expressing CYP3A29 isoenzyme was successfully established, and it may be used for the metabolic study of related drugs.

Objective To discuss the effects of utero placental ischemia on the body and nervous system development in fetal rats Methods By clamping the unilateral uterine artery of the rat, we produced a utero placental ischemia model The opposite uterus of the rat with normal uterine artery supply served as control We compared the body weight, weight of brain, and the expression of growth associated protein 43(GAP 43) mRNA in cerebral tissue by RT PCR in the 13 day (group 1) and 17 day(group 2) old fetal rats respectively Results The body weight and weight of brain in group 1 were 3 2 g and 0 16 g respectively, significantly lower than those of control 1 of 3 6 g and 0 18 g respectively ( P 0 05) However, GAP 43 mRNA in cerebral tissue of the group 2 (1 06) was significantly decreased compared with that of its control(1 21 )( P

OBJECTIVEThis study aims to test the accuracy and precision of iWitness photogrammetry for measuring the facial tissues of mannequin head.

METHODSUnder ideal circumstances, the 3D landmark coordinates were repeatedly obtained from a mannequin head using iWitness photogrammetric system with different parameters, to examine the precision of this system. The differences between the 3D data and their true distance values of mannequin head were computed.

RESULTSOperator error of 3D system in non-zoom and zoom status were 0.20 mm and 0.09 mm, and the difference was significant (P 0.05). Image captured error of 3D system was 0.283 mm, and there was no significant difference compared with the same group of images (P>0.05). Error of 3D systen with recalibration was 0.251 mm, and the difference was not statistically significant compared with image captured error (P>0.05). Good congruence was observed between means derived from the 3D photos and direct anthropometry, with difference ranging from -0.4 mm to +0.4 mm.

CONCLUSIONThis study provides further evidence of the high reliability of iWitness photogrammetry for several craniofacial measurements, including landmarks and inter-landmark distances. The evaluated system can be recommended for the evaluation and documentation of the facial surface.

Anthropometry ; Cephalometry ; Face ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Manikins ; Photogrammetry ; Reproducibility of Results

Objective To discuss the effects of abnormal nutritional supply during pregnancy on the adult insulin and leptin resistance in rats.Methods We established the pregnant rat models given either low protein or high nutrition diet,and normal diet group served as control.There were 12 pregnant rats in each group.After vaginal delivery,the pups birth weight were measured.The randomly selected small for gestational age(SGA)from low protein diet group,large for gestational age(LGA)pups from high nutrition diet group and normal birth weight pups from control group were studied at both 4 weeks and 12 weeks after being born.Each group contained 36 rats.The insulin and leptin level were measured by the method of ELISA,and insulin sensitive index were calculated respectively.Results The pups of mothers served with low protein showed obviously lower birth weight than those mothers with normal diet(P0.05),while the weight of fat around kidney,the ratio of fat and body weight(FW/BW)were(0.36?0.14)g,6.5?0.3,which were higher than those of control (0.19?0.13)g,3.4+O.3(P

Proteins extracted from two varieties of Chinese roses leaves were separated by two- dimensional polyacrylamide gel electrophoresis (2-DE) with immobilized pH gradient (IPG). Many difference proteins were isolated with molecular weights ranging 10-30 kDa and pI5-6. Three proteins of high levels observed in a gel were excised and identified using peptide mass fingerprinting and MS-MS. A summary of the identified proteins and their putative functions are presented. They are identified as eIF-5A、LEA protein and Hsp17. 5. Functions of these proteins in plant tolerance to high temperature were discussed.

OBJECTIVETo study the effect of herbal compound 861 (HB861) on expression and activity of nitric oxide synthase (NOS) in hepatic stellate cells (HSC), and to explore the feasibility of its application in preventing and treating the early portal hypertension.

METHODSHSC of HSC-T6 cell line (1 x 10(5)/ml) were cultured in dish with 95% O2 plus 5% CO2 under 37 degrees C for 24 hrs, then divided into 5 groups, 6 dishes in each group. Group A was the blank control group. To Group B-E, HB861 2 mg/ml, HB861 4 mg/ml, HB861 8 mg/ml, HB861 4 mg/ml + NW-Nitro-L-Arginine Methyl Ester (L-NAME)4 mg/ml were added separately, and continuously cultured for 24 hrs. NOS activity was measured using colorimetry, NO level was determined by nitrate reductase technique. The cells were fixed by 4% paraformaldehyde for 2 hrs for test HSC-T6 iNOS expression by immunocyto-chemical method.

RESULTSHB861 in 2 mg/ml, 4 mg/ml and 8 mg/ml could increase HSC-T6 NOS activity from 1.7 +/- 0.1 to 2.5 +/- 0.3, 3.5 +/- 0.4 and 3.7 +/- 0.9 respectively (P < 0.01), the NO levels in supernatant were increased in parallel from 56.1 +/- 4.8 to 90.7 +/- 4.6, 99.7 +/- 4.1 and 109.0 +/- 2.7 respectively (P < 0.01). L-NAME could not inhibit the effect of HB861 in increasing the synthesis and secretion of NO by activated HSC-T6. Immuno-cyto-chemical study showed that there was iNOS expression in cytoplasm, and which could be increased by HB861.

CONCLUSIONThe activated HSC-T6 showed positive iNOS expression, suggesting it could produce NO. HB861 could markedly increase HSC-T6 iNOS expression and NOS activity, enhance the NO synthesis and secretion, it also could inhibit the contractility of activated HSC by way of increase HSC to secrete NO, so as to lower the resistance in hepatic sinusoid, therefore would play important role in preventing and treating of early portal hypertension.

Animals ; Cells, Cultured ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; enzymology ; Hypertension, Portal ; prevention & control ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; biosynthesis ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective This study aims to analyze if high sensitivity C Reactive Protein (hs-CRP) was a independent risk factor of acute kidney injury(AKI) after A type aortic dissection surgery.Methods Clinical data of the 169 patients who underwent A type acute aortic dissection surgery from February 2009 to October 2010 were collected.Patients without preoperative detection of hs-CRP,patients with preoperative infection and patients diagnosed infection before AKI were excluded.Enrolled patients were divided into AKI group and non-AKI group,and according to using RRT or not,the patients were divided into RRT group and non-RRT group.All the factors were evaluated by means of univariate and multivariate logistic regression analysis to identify relative risk factors of AKI.Results AKI occurred in 95 cases(56.2％),Using RRT in 8 cases (4.7％).hsCRP is an independent risk factor of AKI(OR =0.975,95％ CI 0.952-0.999,P =0.041).hs-CRP and aortic cross clamping time were the independent risk factors of using RRT,The in-hospital mortality was significant difference between RRT group and non-RRT group (P ＜ 0.05).The area under the ROC curve of hs-CRP on RRT diagnosis was 0.733,95％ CI 0.570-0.896,P =0.026.The sensitivity of CRP ＞ 30.42 mg/L warning AKI need RRT was 87.5％,the specificity was 53.4％.Conclusion AKI after A type aortic dissection surgery was a severe complication and RRT associated with in-hospital mortality,hs-CRP was higher in acute aortic dissection patients.The level of hs-CRP and aortic cross clamping time were independent risk factors of AKI and RRT.

In this study, 101 strains of bacteria were isolated from arct ic water and sediment samples. The methanol extracts of the fermented broth prod uced by these strains were screened in vitro for anti-tumor activity on mou se tsFT210 cells using the method of flow cytometry, and screened for antibacter ial activity by the method of paper disk diffusion. The result showed that one strain exhibited anti-tumor activity and eight strains had antibacterial activ ity. The stability of the antibacterial components produced by strain AR084 an d its optimum medium were also studied. The research indicated that arctic bac teria had potential application in pharmaceutics.

OBJECTIVETo culture rat prostate glandular epithelial cells and study their barrier functions in vitro.

METHODSRat prostate glandular epithelial cells were cultured in vitro. The expression of the tight junction protein claudin-1 was determined by immunohistochemistry, the structure and composition of the epithelial cells observed under the inverted microscope and transmission electron microscope. The transepithelial electrical resistances (TEERs) were monitored with the Millicell system. The permeability of the prostate glandular epithelial cells was assessed by the phenol red leakage test.

RESULTSCompact monolayer cell structures were formed in the prostate glandular epithelial cells cultured in vitro. Immunohistochemistry showed the expression of the tight junction protein claudin-1 and transmission electron microscopy confirmed the formation of tight junctions between the adjacent glandular epithelial cells. The TEERs in the cultured prostate glandular epithelial cells reached the peak of about (201.3 ± 3.5) Ω/cm2 on the 8th day. The phenol red leakage test manifested a decreased permeability of the cell layers with the increase of TEERs.

CONCLUSIONThe structure and function of rat prostate glandular epithelial cells are similar to those of brain capillary endothelial cells, retinal capillary endothelial cells, and intestinal epithelial cells. In vitro cultured prostate glandular epithelial cells have the barrier function and can be used as a model for the study of blood prostate barrier in vitro.

Animals ; Cell Membrane Permeability ; Cells, Cultured ; Claudin-1 ; metabolism ; Electric Impedance ; Epithelial Cells ; pathology ; physiology ; ultrastructure ; In Vitro Techniques ; Male ; Microscopy, Electron, Transmission ; Phenolsulfonphthalein ; pharmacokinetics ; Prostate ; metabolism ; pathology ; Rats ; Tight Junctions

AlM:To explore the effective method for the treatment of complicated ocular trauma accompanied with cyclodialysis.METHODS: Sixty - eight cases of complexity ocular trauma with cyclodialysis in different degrees were undergone vitrectomy and ( or ) combined with ciliary body reduction.RESULTS:All patients were followed up for 10 ~36mo (mean 17. 0±5. 7mo). The postoperative visual acuity was better than that of preoperation (P<0. 05). Compared with preoperative, intraocular pressure was significantly increased ( P < 0. 05 ). Successful rate of ciliary body restoration was 91%. CONCLUSlON: For the complicated ocular trauma accompaniedwith cyclodialysis, vitrectomy and ( or ) ciliary body reduction is an effective treatment method.