Objective To detect the expression level of Taurine up?regulated gene 1( TUG1) in the re?nal cell carcinoma and paired paracancerous normal tissues,then explore the relationships between the expression level of TUG1 and clinical characteristics.Methods RNA was Extacted from the resected renal cell carcinoma tissues and paired paracancerous normal tissues of 46 patients respectively,by reverse transcription to get cDNA, the expression level of the TUG1 was detected by RT?qPCR, the relationship between the expression level of TUG1 and the clinicopathological characteristics was analyzed by statistically software. Results The expression of TUG1 in renal cell carcinoma was obviously lower than that in paired paracancerous normal tissues(0.533±0. 027 vs. 1.000±0.298,t=-3.350,P<0.01).The△CT value of Tug1 in 46 cases of renal cell carcinoma after log?arithmic transformation,the minimum value was -5.535,maximum was 3.085,average value was -0.908,with the average of -0.908 as a dividing line,46 cases of renal cell carcinoma with 25 cases (54.34%) were down regulated the expression.The expression level of TUG1 of patients with renal carcinoma have no significant corre?lation with age,sex,type of renal cell carcinoma,TNM staging and UICC/AJCC staging(P>0.05).Conclusion The expression of TUG1 in renal cell carcinoma tissues are down?regulated,which also suggest that it may be re?lated to the tumorigenesis and development of renal cell carcinoma.

Objective To study effect of excessive iodine intake on sodium-iodide symporter(NIS)mRNA and protein expression of breast in lactating rats.Methods60 Wistar rats,having been weaned for one month,were randomly divided into three groups according to their body weights,I.e,①normal iodine(NI,30 rats);②ten fold high iodine(10 HI,15 rats);③one hundred fold high iodine(100 HI,15 rats).Eating food containing iodine of 300μg/L and drinking water of iodine at 5,1845,20 295μg/L,respectively.After fed for 3 months,the rats mated and had offspring,and urine and milk iodine of lactating rats were determined by As-Ce-catalytic spectrophotometric method.Their marmnary glands were sampled at lactation day 10.Then NIS mRNA expression by RT-PCR was determined and NIS protein by immunohistochemistry(SABC)was observed.Results The urine iodine of 10 HI group(3597.5μg/L)and 100HI group(25 404.3μg/L)increased obviously compared with that of NI group(344.7μg/L).The milk iodine of 10HI group(27.1×103μg/L)and 100HI group(191.0×1μg/L)was higher than that of NI group(6.0×103μg/L),but the increased fold of milk iodine was not paralleled with that of urine iodine.Difference of NIS mRNA expression was significant(F=24.19,P＜0.01)among the groups,and the NIS mRNA expression in 10HI(1.250±0.034)and 100HI(1.272±0.039)group were less than that in NI (1.532±0.044)group(P＜0.01).The breast NIS mRNA expression in lactating rats(1.532±0.044)was significantly higher than that in unlactating rats(0.879±0.018,P＜0.01).With the increasing iodine uptake,NIS protein expression decreased.Conclusions The NIS mRNA and protein in rat breasts is down-regulated by excessive iodine intake.So increasing extent of milk iodine concentration is inhibited,which is important to prevent off-spring from getting excessive iodine intake from parental generation.

Human manganese superoxide dismutase (hMn-SOD) cDNA was amplified by RT-PCR from total RNA of human liver cell (L02), and cloned into yeast expression vector pPIC9K containing AOX1 promoter and the alpha-factor signal peptide sequence. The resultant pPIC9K-MnSOD was transformed to P. pastoris GS115, screened for Mut+ carrying multiple copies of hMn-SOD. The positive transformants were fermented in flasks and induced by 0.5% methanol. After 4 days of methanol induction, the expressed hMn-SOD was up to 32% of the total proteins in the supernatant by SDS-PAGE with specific activity of 247.7 u/mg.
Cloning, Molecular ; DNA, Complementary ; biosynthesis ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Superoxide Dismutase ; biosynthesis ; genetics

Objective To analyze the clinical effect of pedicle screw fixation in the treatment of multi-segmental lumbar fracture and dislocation under the guidance of visualization technique. Methods A total of 21 patients with multi-segmental lumbar fracture and dislocation were selected from November 2012 to November 2013. Before the screw implantation, the structure of bilateral pedicle was observed through Mimics software and the implantation parameters were measured. The position of pedicle screws by postoperative CT scan, operation time, and the satisfaction of the patients were assessed. The percentages of anterior vertebral height and Cobb′s angle were measured before operation, 2 weeks and 8 months after operation. Results All patients were satisfied with informed consent score and the way of pedicle screw and the selection of plant were more reasonable. With better screw position, shorter operative time and less blood loss and adverse reactions, pedicle screw fixation achieved good effect. Conclusion With high security and considerable clinical value, pedicle screw fixation in the treatment of multi-segmental lumbar fracture and dislocation under the guidance of visualization technique has exact and good effecct.

Objective To determine the changes of cerebrospinal fluid(CSF) ?-endorphin(?-EP) and C-reactive protein(CRP) in children with central nervous system(CNS) infection.Methods Sixty-five children suffered from CNS infection were determined the plasma and CSF ?-EP and CRP concentration during the acute and recovering stage with radioimmunoassay, which included 48 viral encephalitis, 12 purulent meningitis and 5 tuberculou meningitis,and 24 non-CNS disease children were as control group.Results The concentrations of plasma and CSF ?-EP of every experimental group were obviously higher than those of control group during the early stage of CNS infection and these were obviously lower during the recovering stage. The serum concentration of CRP during acute stage was significantly higher than that during recovering stage. No change of serum and CSF CRP concentration was determined during either the acute or recovering stage in the other two experimental groups.Conclusions Determining the plasma and CSF ?-EP is mea-(ningful) in early diagnosis of CNS infection,and determining the serum CRP at the same time may be helpful in differentiating septic and inseptic infection.

Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.
Cell Wall ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; analysis ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Powders ; Quality Control

Objective Take alkaline ashing method as golden standard to explore the accuracy of chloric acid digestion method in determination of human milk iodine. Methods Sixty one breast milk samples collected in Hexi district of Tianjin was measured by the method for determination of iodine in foodstuff by As3+-Ce4+ catalytic spectrophotometry (referred to as the alkaline ashing method) published in 2008 and the method for determination of iodine in urine by As3+-Ce4+ catalytic spectrophotometry(referred to as acid digestion) published in 1999, respectively. were highly correlated(r = 0.960, t = 26.3, P < 0.01), and the regression equation was (Y) = - 28.1 + 0.808X, in which X was independent variable, that is the results of alkaline ashing method; (Y) was dependent variable, that is the estimated data of chloric acid digestion method. The average difference of the results measured by the two methods was 68.3 μg/L, and the results from chloric acid digestion was 38.9% which lower than that of alkaline samples were diluted by 3,4 and 5-fold and then digested by chloric acid, the liquid clarification rates were 80.3% ashing and chloric acid digestion method were, respectively, 165.4, 110.0 μg/L. Conclusions Compared with alkaline ashing method, the results determined by chloric acid digestion method are significantly lower. It is suggested that there are systemic errors in chloric acid digestion method, which means that alkaline ashing method can not be replaced by the chloric acid digestion method.

Objective To observe the effect of human β-defensin-3 (HBD-3) on proliferation and the secretion of prostaglandin E2(PGE2) and matrix metalloproteinase-1 (MMP-1) in human gingival fibroblasts(HGF).Methods The HGF were cultured with tissue-explant method and the fourth-generation HGF were plated in 96-well plate.All groups except the control group were treated with different concentrations of HBD-3 for 7 days.Then the HGF proliferation was evaluated with methyl thiazolyl tetrazolium(MTT) colorimetry and the secretions of PGE2 and MMP-1 at the 12th hours of each group were detected by enzyme-linked immunosorbent assay(ELISA).Results The result of MTT dynamic monitoring showed that the amount of HGF increased with time in all groups in concentration dependent manner.ELISA showed that the secretions of PGE2 and MMP-1 in 1.0 mg/L HBD-3 group were (350.56 ± 63.96) ng/L and (13.22 ±0.59) μg/L,significantly higher than those in the control group and 10.0 mg/L HBD-3 group (P ＜ 0.05).Conclusions HBD-3 promoted the proliferation of HGF.The low concentration of HBD-3 may play a role in immunoregulation through increasing the secretions of PGE2 and MMP-1.

ObjectiveTo summarize the operative experiences and postoperative complications of pediatric liver transplantation (LT).MethodsClinical data of 88 child patients undergoing LT in West China Hospital, Sichuan University between January 2000 and October 2015 were retrospectively analyzed. Among the patients, 38 were males and 50 were females, with the age ranging from 3 months to 17 years and 7 months old and the median of 2 years and 3 months old. Seventy-seven cases underwent living donor LT and 11 underwent cadaver LT. Thirty-nine cases underwent left lobe transplantation and 49 underwent left lateral lobe transplantation. The main immunosuppressive regimen after operation was adrenocortical hormone + tacrolimus (FK506) or cyclosporin. And heparin was used after operation for anticoagulation. The informed consents of all patients and their families were obtained and the local ethical committee approval was received. The intraoperative conditions and postoperative complications of the patients were observed.ResultsThe average operation time was (7.5±2.4) h, the median intraoperative blood loss was 475(100-2 000) ml, the transfusion of red blood cells was 1.8(0-6.5) U, the warm ischemia time was 2.5(1.0-10.0) min, the cold ischemia time was 188(110-590) min and the inferior vena cava occlusion time was 75(35-170) min. Forty-nine cases developed early complications after operation, and the incidence of vascular complications was 11%(10/88), including 4 cases of hepatic artery thrombosis, 5 of portal vein thrombosis and 1 of hepatic vein stenosis. Sixteen cases developed late complications, and the incidence of biliary complications was 12% (11/88), including 9 cases of cholangitis, 1 of bile leakage and 1 of bile duct stricture. The 1-, 3-year survival rate after LT was respectively 81% and 75%.Conclusions The success rate and postoperative survival rate of pediatric LT have increased signiifcantly in recent years. However, the incidence of postoperative vascular and biliary complications are still high. Thus, the operation procedures should be kept improving, and the immunosuppressant and anticoagulant therapy should be individually adjusted in order to obtain better efifcacy.

OBJECTIVETo investigate the effect of N-desulfated heparin on tumor metastasis, tumor angiogenesis and basic fibroblast growth factor(bFGF) gene expression of orthotopically implanted human gastric carcinoma in NOD-SCID mice.

METHODSHuman gastric cancer SGC-7901 tissues were orthotopically implanted into the stomach of the NOD-SCID mice. Twenty mice were randomly divided into two groups which received either intravenous injection of 0.9% NaCl solution(0.9%NaCl solution group) or 10 mg/kg N-desulfated heparin (N-desulfated heparin group) twice a week for three weeks. Mice were sacrificed six weeks after tumor implantation. Tissues from stomach and other organs were obtained for histopathological evaluation. The intratumoral microvessel density (MVD) in tumor was evaluated immunohistochemically. Real time PCR was used to detect bFGF mRNA expression.

RESULTSThe tumor metastasis rates were 9/10 in 0.9% NaCl solution group and 2/10 in N-desulfated heparin group(P<0.05).MVD was 9.1+/-3.4 in 0.9% NaCl solution group and 4.7+/-1.8 in N-desulfated heparin group (t=3.617,P<0.05). bFGF mRNA expression was lower in N-desulfated heparin group(2.60+/-0.56%)than that in 0.9% NaCl solution group(30.65+/-6.84%).

CONCLUSIONN-desulfated heparin can inhibit the metastasis of gastric cancer through inhibiting tumor bFGF gene expression and tumor angiogenesis with no obvious anticoagulant activity.

Animals ; Fibroblast Growth Factor 2 ; genetics ; Gene Expression Regulation, Neoplastic ; drug effects ; genetics ; Heparin ; analogs & derivatives ; pharmacology ; therapeutic use ; Humans ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Neoplasm Metastasis ; Neoplasm Transplantation ; Neovascularization, Pathologic ; drug therapy ; Polymerase Chain Reaction ; RNA, Messenger ; genetics ; metabolism ; Stomach Neoplasms ; blood supply ; drug therapy ; genetics