Objective To study the combined effect of interleukin-21 gene transfer and ionizing radiation on the growth of cervical carcinoma HeLa cells.Methods Previously constructed Ad-IL-21 gene was amplified by infecting 293A cells and the titer was measured by TCID50 method. HeLa cells were transfected with Ad-1L-21 and then irradiated with 6 Gy 137Cs γ-rays.The cells were divided into 5 groups,including blank control,Ad-LaeZ group,Ad-IL-21 group,radiation group and Ad-IL-21 combined with radiation group (combination group).The cell growth,cell cycle,apoptosis,and the expressions of IL-21 gene and protein in HeLa cells were detected.Results Ad-IL-21 was successfully amplified and the titer of Ad-11.-21 was 9 × 1010 pfu/ml.Compared with Ad-IL-21 group and radiation group,the cell growth of combination group was significantly inhibited at 96 h after transfection ( F =85.26,72.98,P ＜ 0.05 ).The cells in combination group were arrested in G1 phase and decreased at S phase( F =36.69,34.83,P ＜ 0.05),while the cellular apoptosis increased markedly ( F =28.23,25.57,P ＜ O.05 ). The gene expression of 1L-21 in the combination group was 1.54- and 2.43-fold of Ad-IL-21 group and blank control group,respectively (F=22.31,36.65, P ＜ 0.05 ), while the protein expression of IL-21 in the combination group was 1.62-fold and 2.31-fold of Ad-IL-21 group and blank control group,respectively ( F =27.36,35.86,P ＜ 0.05 ).Conclusions Ad-IL-21 gene transfection combined with radiation has synergic effect on the inhibition of cervical carcinoma cell growth.

Objective To investigate the therapeutic effect of post-traumatic complex posterior urethral stricture in the male patients.Methods Clinical data of 479 male patients with post-traumatic complex posterior urethral stricture were reviewed.One-stage resection of the stenosis and end-to-end anastomosis was performed in 422 patients and scrotal flap with blood pedicle posterior urethroplasty in 57.Results The mean operation time was 115 minutes(range,90-140 minutes).The mean blood loss was 225 ml(range,100-300 ml).No intraoperative blood transfusion was needed.The mean follow-up time was 15 months(range,12-24 months).Among the 422 patients performed end-to-end anastomosis,386 patients had good voiding and 36 had dysuria because of the formation of anastomotic stoma valve(21 patients)or stricture ring(15 patients).The problem was resolved by transurethral valve/stricture ring resection.Among 57 patients undergone posterior urethroplasty,45 patients had good voiding nine patients were found with anterior urethra-skin tube anastomotic stoma stricture,of which four patients were treated by urethral dilatation and five by endourethrotomy; three patients were found with posterior urethra-skin tube anastomotic stoma stricture,of which one patient was treated by urethral dilation and two by endourethrotomy.Conclusions One-stage resection of the stenosis and end-to-end anastomosis is the main treatment for post-traumatic complex posterior urethral stricture.If the condition of the patients does not allow the end-to-end anastomosis,posterior urethroplasty can be an alternative.

OBJECTIVETo understand the current situations of saturnism and blood lead levels of children resided in village and circumjacent areas, and to know its relations with sex, age and other factors on children' s health as to providing some evidences for prevention and control.

METHODSAn epidemiological survey was conducted for finding out the pollution sources and for a better understanding of the surrounding environment. All 221 children under 14 years old, from the lead pollution villages and surrounding establishments were enrolled, and their blood lead levels were detected by graphite atomizer absorption spectrophotometer method. Symptoms of the saturnism were investigated through a standardized questionnaire. SPSS13.0 software was administrated for data analysis.

RESULTSHigh blood lead level identification rate was 66.06% (146/221), and saturnism rate 32.13% (71/221). The children's blood lead levels among group 1, group 2, group 3 in this village and jade factory were (161.20 +/- 32.94), (176.60 +/- 43.62), (258.00 +/-106.08) and (238.01 +/- 55.20) microg/L respectively and the significant differences were observed through Kruskal-Wallis test (chi2 = 51.84, df= 3, P<0. 01). The blood lead levels of children from group 3 in this village and the jade factory were higher than those of other two groups. No correlation was found between children's age and blood lead level (r = 0.10, P = 0.13). There was a significant difference in blood lead levels between boys and girls (t' = 3.83, P<0.01). With the children's blood lead levels rising, the occurrence rate of main saturnism symptoms was significantly increased. This survey suggested that the pollution source was a coarse lead smelter.

CONCLUSIONThe blood lead level should ke overwhelmingly increased among children who live nearby the higher level of lead blood, that living nearby the lead smeltery,might result in stautnism and negative effect on children's healthy.

Child ; Environmental Pollution ; analysis ; Female ; Humans ; Industrial Waste ; Lead ; blood ; Lead Poisoning ; epidemiology ; Male ; Metallurgy

OBJECTIVETo develop a small-caliber vascular grafts and study its morphologies, mechanical properties and biocompatibility.

METHODSThe effects of electrospinning conditions on the microstructure and porosity of the resulting scaffolds were investigated for preparation of a small-caliber (4 mm) polyurethane vascular grafts with optimum microstructures and mechanical properties. The mechanical properties and biocompatibility of the prepared grafts were evaluated.

RESULTSThe polyurethane vascular grafts showed a three-dimensional reticular structure consisting of nanofibers, with an average porosity of (51.48∓4.47)% and tensile strength of 5.85 ∓ 0.62 MPa. The grafts provided a better long-term support than e-PTFE graft for endothelial cell growth and endothelialization.

CONCLUSIONThe polyurethane vascular prosthesis possessed favorable microstructures, excellent mechanical properties and good biocompatibility for potential clinical application.

Biocompatible Materials ; chemistry ; Blood Vessel Prosthesis ; Cell Adhesion ; Humans ; Materials Testing ; Mechanical Phenomena ; Polyurethanes ; chemistry ; Porosity ; Prosthesis Design ; Tensile Strength

A rice pse(t) (premature senescence, tentatively) mutant line, was isolated from 4,500 independent T-DNA inserted transgenic lines. The symptoms of premature senescence appeared more severely than those of the control plants (Zhonghua 11, japonica) at the last development stage. To characterize the mutant and provide basic information on the candidate genes by mapping to a physical region of 220-kb, experiments were carried out in two phytotrons under controlled temperature of 24 degrees C and 28 degrees C, respectively. The content of chlorophyll, soluble protein and MDA (malondialdehyde), net photosynthesis, the antioxidant enzyme activities of SOD (superoxide dismuase) (EC 1.15.1.1) and POD (peroxidase) (EC 1.11.1.7) and the peptidase activities of leaves were measured from top to bottom according to the leaf positions at the flowering stage. Compared with the control plant, the mutant showed the following characteristics: (1) Higher net photosynthesis rate (P(n)) appeared in the 1st and 2nd leaves, contents of chlorophyll and soluble protein were also higher in the 1st leaf; (2) The activities of SOD, POD and peptidase were higher according to the leaf position from top to bottom; (3) The symptom of premature senescence was accelerated in the mutant at 28 degrees C treatment. The MDA content and the SOD and POD activities between the 24 degrees C and 28 degrees C treatment mutants were not significantly different. Content of chlorophyll and soluble protein of leaves mutant decreased rapidly at 28 degrees C treatment. The results show that pse(t) is sensitive to high temperature. The probable function of PSE(T) is discussed.
Aging ; physiology ; Antioxidants ; metabolism ; Apoptosis ; physiology ; Mutation ; Oryza ; classification ; genetics ; growth & development ; Photosynthesis ; physiology ; Plant Leaves ; classification ; genetics ; growth & development ; Plant Proteins ; genetics ; metabolism

OBJECTIVETo investigate the protective effect of manganese superoxide dismutase (MnSOD) gene transfer to small intestinal epithelial cells from radiation injury.

METHODSHerpes simplex virus (HSV) vector containing both the human MnSOD and GFP genes was introduced into mouse small intestine. Expression of MnSOD by the intestinal villi was confirmed by nested RT-PCR, immunofluorescence and enzyme activity assay. Mice were then given various doses of irradiation over the abdomen. The height of intestinal villi was measured on histopathology sections by SZ-PT optical system before irradiation, 24 h and 72 h post-irradiation. All comparisons were performed by one-way analysis of variance using the SPSS statistical software to analyze the significance between groups.

RESULTSNested RT-PCR, immunofluorescence and enzyme activity assay of MnSOD demonstrated overexpression and increased activity of MnSOD in the inoculated intestine of mice. Control (sham inoculated) irradiated mice showed decreased villi height by 40.1%-59.3% on day 1 and 44.2%-65.1% on day 3 (7.5-15 Gy). Treatment of mice with HSV-MnSOD prior to radiation led to statistically significant radioprotection of the small bowel with mean villi height decreased by only 3.1%-12.4% on day 1 and 6.3%-29.1% on day 3.

CONCLUSIONThe results demonstrate that overexpression of human MnSOD via a replication defective herpes simplex viral vector is an effective method to protect the small intestine from damage caused by ionizing radiation.

Animals ; Epithelial Cells ; metabolism ; Genetic Therapy ; Genetic Vectors ; Intestine, Small ; metabolism ; Mice ; Radiation Injuries, Experimental ; prevention & control ; Simplexvirus ; genetics ; Superoxide Dismutase ; genetics ; Transfection

OBJECTIVETo explore the synergetic effect of norcantharidin (NCTD) and adriamycin (ADR) on the proliferation and apoptosis of multiple myeloma (MM) cells.

METHODSHuman MM cell line U266 cells were treated with NCTD alone (10 µmol/L) or in combination with ADR (0.25 µmol/L). MTT and Annexin V/PI staining were used to determine cell viability and apoptosis. The protein expression of nuclear factor-κB P65 (NF-κB P65), phosphorylated NF-κB p65 (p-NF-κB p65), NF-κB P65 inhibitor IκBα, phosphorylated IκBα (p-IκBα), survivin, Bcl-2 and Bax were determined by Western blot. Immunohistochemistry was used to determine the expression of vascular endothelial growth factor (VEGF).

RESULTS(1) NCTD potentiated the cytotoxicity and pro-apoptotic effects induced by ADR. The combination of NCTD and ADR had synergistic anti-proliferation effect. (2) Combination of ADR and NCTD downregulated the expression of nuclear NF-κB P65 and cytoplasm p-IκBα induced by ADR. The expression of nuclear NF-κB P65 and cytoplasm p-IκBα decreased from 2.08 ± 0.29 and 0.39 ± 0.07 to 0.48 ± 0.08 and 0.02 ± 0.01 respectively, while the expression of the cytoplasm NF-κB P65 and IκBα were unchanged in the ADR alone group and the combined group. (3) The expression of survivin and bcl-2 decreased from 0.31 ± 0.05 and 0.23 ± 0.05 to 0.03 ± 0.02 and 0.05 ± 0.02, while the expression of Bax increased from 0.46 ± 0.06 to 0.62 ± 0.08 respectively in ADR alone group and combined group. (4) The positive rate of VEGF in ADR group and combination group were (44.6 ± 4.4)% and (27.0 ± 2.1)% respectively, indicating that NCTD could potentiate the inhibition effect on VEGF induced by ADR.

CONCLUSIONSThe results suggest that NCTD can potentialize the chemosensitivity of multiple myeloma cells to ADR through regulating NF-κB/IκBα signaling pathway and NF-κB-regulated gene products including survivin, Bcl-2, Bax and VEGF.

Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Doxorubicin ; pharmacology ; Humans ; I-kappa B Proteins ; metabolism ; Inhibitor of Apoptosis Proteins ; metabolism ; Multiple Myeloma ; metabolism ; NF-KappaB Inhibitor alpha ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction ; drug effects ; Transcription Factor RelA ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the effects of Apo A1 and B gene polymorphism on avascular necrosis of the femoral head (ANFH) in north Chinese Han population.

METHODSPolymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) technique was used in samples of 143 cases with documented ANFH and 92 healthy control matched by age and sex individuals selected from north Chinese Han nationality. The studied loci include promoter region (-75bp) and the intron 1 (+83 bp) of Apo A1 gene, Eco RI,Xba I of Apo B gene; polymerase chain reaction was used to study 3'-VNTR of Apo B gene.

RESULTSAt -75 bp in promoter, the frequency of A/A genotype in ANFH group was significantly higher than that in control group (P < 0.01), while the frequency of G/A genotype in ANFH group was significantly lower than that in control group (P < 0.01). No difference was found in the frequency of genotype at +83bp in intron 1 of Apo A1 gene, Eco RI, Xba I and 3'-VNTR loci of Apo B gene.

CONCLUSIONApolipoprotein A1 gene A/A substitution at position -75 in promotor is associated with ANFH, the mutation may be one of the sensitive genes of ANFH, first reported inside and abroad. But no evident relationship was found between gene polymorphism of +75 bp loci of Apo A1 gene, Eco RI loci of Apo B gene, Xba I loci of Apo B gene or 3'-VNTR of Apo B gene and ANFH.

Adult ; Apolipoprotein A-I ; genetics ; Apolipoproteins B ; genetics ; Case-Control Studies ; China ; Electrophoresis ; Female ; Femur Head Necrosis ; genetics ; Gene Frequency ; Genotype ; Humans ; Male ; Polymorphism, Genetic

This study was aimed at evaluating the antioxidant activities of methanol extract of Thamnolia vermicularis. The antioxidant activity, reducing power, superoxide anion radical scavenging and free radical scavenging activities were studied. The antioxidant activity of the extract correlated with its concentration (0.2~2 mg/ml) in the reaction mixtures containing linoleic acid. Upto 67% of lipid peroxidation was inhibited by 2 mg/ml of the lichen extract. The extract showed strong free radical scavenging activity similar to that of BHA (positive control) in a manner of concentration dependent. The lichen extract also showed moderate effects on superoxide anoin scavenging activity and reducing power, which was not so effective as that of Quercetin and BHA used as positive controls. This study suggests that T. vermicularis lichen can be used as a novel source of natural antioxidant.
Butylated Hydroxyanisole ; Lichens* ; Linoleic Acid ; Lipid Peroxidation ; Methanol ; Quercetin ; Superoxides

OBJECTIVETo observe the expressions of the substance P (SP) mRNA and neurokinin-1 receptor (NK-1R) in the posterior horn of the L5 - S2 spinal cord in the rat model of chronic prostatitis pain, and to investigate the changes in the activation of astrocytes and influence of SP on this activation in rat spinal cord astrocytes cultured in vitro.

METHODSThe rat model of chronic prostatitis pain was established by injection of complete Freund's adjuvant (CFA) and assessed by the tail flick threshold test, the control rats injected with sodium chloride and all observed at 0, 14 and 28 days. Changes in the expressions of SP mRNA, NK-1R, glial fibrillary acidic protein (GFAP), tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS) in the posterior horn of the L5 - S2 spinal cord were detected by RT-PCR and Western blot. Rat spinal cord astrocytes were cultured in vitro and divided into a control group, cultured with ITS cell culture fluid, and two experiment groups, with Group 1 stimulated with SP at the concentration of 10(-9) - 10(-6) mol/L for 12 hours followed by determination of the expressions of TNF-alpha, IL-1beta, NO and NOS by ELISA and nitrate reductase and colorimetric methods, and Group 2 at 10(-7) mol/L for 0, 24, 48 and 72 hours followed by detection of the GFAP expression by Western blot.

RESULTSThe expressions of SP mRNA, NK-1 R, GFAP, TNF-alpha and iNOS in the posterior horn of the L5 - S2 spinal cord were obviously higher in the rat prostatitis pain models than in the controls, successively higher at 28 than at 14 and 0 d (P < 0.01), and so was the expression of GFAP at 28 than at 14 d in the experiment groups (P < 0.05). SP induced a gradual increase at 10(-7) mol/L in the expression of GFAP in the spinal cord astrocytes at 0 -72 h, significantly different from that of the control group (P < 0.01), and it promoted the excretion of TNF-alpha and IL-1beta and the activity of NO and NOS at 10(-9) - 10(-6) mol/L at 12 h in a concentration-dependent manner, with marked differences between the experiment and control groups (P < 0.01, P < 0.05). But a decreased excretion of IL-1 beta was observed in the 10(-6) mol/L group, though with no significant difference from the control (P > 0.05).

CONCLUSIONChronic prostatitis pain could upregulate the expressions of the excitatory transmitter SP and receptor in the L5 - S2 spinal cord, and result in the activation of astrocytes and increased excretion of proinflammatory cytokines, which may be associated with the persistence and generalization of prostatitis pain.

Animals ; Astrocytes ; metabolism ; Chronic Disease ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Pain ; metabolism ; Prostatitis ; metabolism ; Rats ; Receptors, Neurokinin-1 ; metabolism ; Spinal Cord ; cytology ; metabolism ; pathology ; Substance P ; metabolism