The use of optimal regulatory sequences for simultaneous expression of the transgenes might play a significant role in engineering plants with increased disease and insect resistance.The plant expression vector pOMS-GUS,which contained the GUS gene under the control of a chimeric promoter based upon the mannopine synthase(mas)promoter and the octopine synthase(ocs)enhancer,was constructed.Used as control,another vector pMAS-GUS,carried the GUS gene driven by only the mas promoter.The two vectors were introduced into tobacco plants by Agrobacterium-mediated transformation.Fluorometric assays for GUS activity and reverse transcription-polymerase chain reaction(RT-PCR)analysis revealed that GUS gene expressed weakly with untreated transgenic tobacco while the level of GUS activity increased steadily after 1 h subjected to wounding.The expression of the mas and ocs/mas promoters was induced a further 1.8-fold and 5.7-fold,respectively.SA(1 mmol/L)or MJ(250 ?mol/L)treatment also caused a large induction of the ocs/mas chimeric promoter;And the application of SA in combination with MJ(1 mmol/LSA & 250 ?mol/L MJ)produced an additive effect that exceeded the wounding response.The results showed that the ocs/mas chimeric promoter is a strong inducible promoter that can be activated by various stresses.The chimeric promoter should have utility in development of disease and insect resistant transgenic crops.

Angiopoietins ; metabolism ; physiology ; Animals ; Cell Proliferation ; Endothelial Cells ; cytology ; physiology ; Humans ; Neoplasms ; blood supply ; Neovascularization, Pathologic ; physiopathology ; Neovascularization, Physiologic ; physiology ; Pericytes ; cytology ; metabolism ; physiology ; Proto-Oncogene Proteins c-sis ; metabolism ; physiology ; Receptor, Platelet-Derived Growth Factor beta ; metabolism ; physiology ; Receptor, TIE-2 ; metabolism ; physiology ; Signal Transduction

Adult ; Benzamides ; pharmacology ; Humans ; Hypereosinophilic Syndrome ; drug therapy ; genetics ; Imatinib Mesylate ; Male ; Mutation ; Oncogene Proteins, Fusion ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; Receptor, Platelet-Derived Growth Factor alpha ; genetics ; mRNA Cleavage and Polyadenylation Factors

OBJECTIVETo examine the reliability and validity of the 66-item version on the gross motor function measure (GMFM-66) to assess the gross motor functions of children below 3 years oldwith cerebral palsy.

METHODS298 valid samples were obtained from 171 children with cerebral palsy (male 126, female 45 with mean age 19 months, age range 3-36 months) measured with GMFM-88. Then a 73-item version of GMFM (GMFM-73) special for these children was obtained by Rasch analysis. Both GMFM-66 and GMFM-73 scores of to each sample were obtained. The reliability and validity of GMFM-66 were evaluated through analyzing the correlation between the scores and between the changed scores of these two GMFM versions. The relative precision of GMFM-73 versus GMFM-66 was also analyzed.

RESULTSSignificant correlations were found between the scores and between the changed scores of these two versions of GMFM. A 14% less gain in relative precision was achieved when using GMFM-73 versus GMFM-66.

CONCLUSIONResults indicated that the GMFM-66 had good reliability and validity in assessing the gross motor functions of children below 3 years old with cerebral palsy.

Cerebral Palsy ; physiopathology ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Motor Skills ; classification ; Reproducibility of Results ; Surveys and Questionnaires

Objective To investigate the feasibility of complicate myomectomy.Methods Six- seven patients with complicated uterine myomas undergoing laparoscopic myomectomy were retrospectively analyzed.The myomectomy was done using ureteral infravision imaging system or/and with self-made myoma segregate-stick.Among the total,there are 29 cases of multiple myomas(the number of myomas≥5)and 23 cases of single myoma(the diameter of myoma≥7cm,including 19 cases of intramural myoma,4 cases of subserous myoma),6 cases of myoma of broad ligament of uterus and 9 cases of cervical myoma.Results All cases were performed successfully laparoscopically.No intra-operative laparotomy or complications occurred.The average operating time and blood loss were(114?32)min and(114?78)ml respectively. The average time of hospital stay was 5.1 d.The average operating time and blood loss in the group (including multiple myoma group,intramural myoma group and cervical myoma)were significantly exceeded the other groups(including myoma of broad ligament of uterus group and subserous myoma group). Conclusions Laparoscopic complicate myomectomy can be performed and the operation indication is enlarged using Ureteral Infravision Imaging System.Advancement in surgical instruments and expert operating skills are the key to operation success.

Background::The use of microRNAs in the therapy of kidney disease is hampered by the difficulties in their effective delivery. Microvesicles (MVs) are known as natural carriers of small RNAs. Our prior research has demonstrated that MVs isolated from mesenchymal stem cells (MSCs) are capable of attenuating kidney injuries induced by unilateral ureteral obstruction and 5/6 sub-total nephrectomy in mice. The present study aimed to evaluate the effects of miR-34a-5p (miR-34a)-modified MSC-MVs on transforming growth factor (TGF)-β1-induced fibrosis and apoptosis in vitro. Methods::Bone marrow MSCs were modified by lentiviruses over-expressing miR-34a, from which MVs were collected for the treatment of human Kidney-2 (HK-2) renal tubular cells exposed to TGF-β1 (6 ng/mL). The survival of HK-2 cells was determined using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and Annexin V-Light 650/propidium iodide (PI) assays. The expression levels of epithelial markers (tight junction protein 1 [TJP1] and E-cadherin) and mesenchymal markers (smooth muscle actin alpha (α-SMA) and fibronectin) in HK-2 cells were measured using Western blot analysis and an immunofluorescence assay. In addition, changes in Notch-1/Jagged-1 signaling were analyzed using Western blotting. Data were analyzed using a Student’s t test or one-way analysis of variance. Results::MiR-34a expression increased three-fold in MVs generated by miR-34a-modified MSCs compared with that expressed in control MVs ( P < 0.01, t= 16.55). In HK-2 cells, TJP1 and E-cadherin levels decreased to 31% and 37% after treatment with TGF-β1, respectively, and were restored to 62% and 70% by miR-34a-enriched MSC-MVs, respectively. The expression of α-SMA and fibronectin increased by 3.9- and 5.0-fold following TGF-β1 treatment, and decreased to 2.0- and 1.7-fold after treatment of HK-2 cells with miR-34a-enriched MSC-MVs. The effects of miR-34a-enriched MSC-MVs on epithelial-mesenchymal transition (EMT) markers were stronger than control MSC-MVs. The effects of miR-34a-enriched MSC-MVs on these EMT markers were stronger than control MSC-MVs. Notch-1 receptor and Jagged-1 ligand, two major molecules of Notch signaling pathway, are predicted targets of miR-34a. It was further observed that elevation of Notch-1 and Jagged-1 induced by TGF-β1 was inhibited by miR-34a-enriched MSC-MVs. In addition, TGF-β1 exposure also induced apoptosis in HK-2 cells. Although miR-34a-mofidied MSC-MVs were able to inhibit TGF-β1-triggered apoptosis in HK-2 cells, the effects were less significant than control MSC-MVs (control:TGF-β1 :miR-nc-MV:miR-34a-MV = 1.3:0.6:1.1:0.9 for MTT assay, 1.8%:23.3%:9.4%:17.4% for apoptosis assay). This phenomenon may be the result of the pro-apoptotic effects of miR-34a. Conclusions::The present study demonstrated that miR-34a-over-expressing MSC-MVs inhibit EMT induced by pro-fibrotic TGF-β1 in renal tubular epithelial cells, possibly through inhibition of the Jagged-1/Notch-1 pathway. Genetic modification of MSC-MVs with an anti-fibrotic molecule may represent a novel strategy for the treatment of renal injuries.

OBJECTIVETo study mechanism of antitumor activity of Sargassum Fusiforme Polysaccharide (SFPS).

METHODThe effect on cell cycle and apoptosis was studied with flow cytometry (FCM). Intracellular calcium concentration [Ca2+]i was marked with Fluo-3/AM and measured with laser scanner confocal microscope (LSCM).

RESULTSFPS inhibited G0/G1 stage SGC-7901 from entering to S stage and increased APO%. The [Ca2+]i showed a transient rise and return to the original level. The concentration could be raised again by administering CaCl2.

CONCLUSIONThe antitumor effect of SFPS seems to be accomplished through the apoptosis associated with the increase in intracellular calcium concentration. Intracellular stores release the calcium during its action.

Antineoplastic Agents ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Calcium ; metabolism ; Calcium Chloride ; pharmacology ; Cell Cycle ; drug effects ; Humans ; Polysaccharides ; isolation & purification ; pharmacology ; Sargassum ; chemistry ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Cells, Cultured

Adult ; Aged ; Aged, 80 and over ; Female ; Hepatic Encephalopathy ; diagnosis ; Humans ; Male ; Middle Aged ; Neuropsychological Tests ; Psychometrics ; methods ; Young Adult

OBJECTIVETo investigate the possible association of single nucleotide polymorphism (SNP) at the 41657C/T position and 4234G/C position of X-ray repair cross-complementing gene 2 (XRCC2) with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA) in a population of high incidence region, Ci county and She county of Hebei.

METHODSThe genotypes of XRCC2 41657C/T and 4234G/C SNPs were detected by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis in 330 ESCC patients, 254 GCA patients and 629 healthy controls.

RESULTSThe genotype frequency of XRCC2 41657C/T in ESCC patients (67.8%, 26.4% and 5.8%) was significantly different from that in controls (68.8%, 28.8% and 2.4%; chi square was 7.43, P was 0.02). Compared with CC genotype, TT genotype significantly increased the risk of developing ESCC (OR=2.12, 95%CI: 1.03-4.35). The genotype (59.9%, 35.8% and 4.3%) and allelotype distributions ofXRCC2 41657C/T in GCA patients were significantly different from that in controls (chi square was 7.46 and 7.23, P was 0.02 and 0.01). Compared with CC genotype, CT genotype significantly increased the risk of developing GCA (OR=1.38, 95%CI: 1.01-1.89). The genotype and allelotype distributions of the 4234G/C SNPs in ESCC and GCA patients were not significantly different from that in controls (all P values were above 0.05). Compared with GG genotype, the CG and CC genotype of XRCC2 4234G/C did not show significant effect on the risk of developing ESCC and GCA. When the two XRCC2 SNPs were combined analyzed, the haplotype distribution in GCA patients was significantly different from that in controls (chi square was 13.28, P was less than 0.01). Compared with 41657C/4234G haplotype, 41657C/4234C and 41657T/4234G haplotypes significantly increased the risk of developing GCA (OR were 1.44 and 1.55, 95%CI were 1.06-1.95 and 1.18-2.02, respectively).

CONCLUSIONIn high incidence region of Hebei province, we conclude that XRCC2 41657C/T polymorphism has a potential to be a susceptibility factor for ESCC and GCA while XRCC2 4234G/C polymorphism may not provide a useful marker to predict susceptibility to ESCC and GCA. However, the 41657C/4234C and 41657T/4234G haplotypes might increase the risk of developing GCA.

Adenocarcinoma ; genetics ; Alleles ; Asian Continental Ancestry Group ; genetics ; Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; China ; DNA-Binding Proteins ; genetics ; Esophageal Neoplasms ; genetics ; Female ; Genetic Predisposition to Disease ; Haplotypes ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Stomach Neoplasms ; genetics

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; genetics ; physiology ; Cells, Cultured ; Diabetes Mellitus ; therapy ; Immunohistochemistry ; Islets of Langerhans ; cytology ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Polymerase Chain Reaction ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley