Objective To develop a sensitive,quick and convenient comet assay with Viciafaba root cells,so as to provide the reference for biomonitoring in situ.Methods The K_2CrO_7 and H_2O_2 were used as the positive control,the Viciafaba root cells isolated with mechanical and enzyme treatment respectively were exposed to the different concentrations of K_2CrO_7(1 nmol/L,10 nmol/L,100 nmol/L,1000 nmol/L)and H202(25 ? mol/L,50 ? mol/L,100 ?mol/L,200 ?mol/L).The effect of DNA damage was detected by using comet assay.Results The yield of Viciafaba root cells isolated from enzyme treatment were much higher than that from mechanical treatment.K_2CrO_7and H_2O_2 could cause DNA damage in Viciafaba root cells,there was significant difference between exposure group and control group(P

[ ABSTRACT] AIM: To explore whether autophagy is involved in the excessive death of renal tubular epithelial cells in subtotal nephrectomy ( SNx) rats and the relationship between autophagy and necroptosis in the kidney of SNx rats. METHODS:Male Sprague-Dawley rats were randomly assigned to control group ( n=6 ) and SNx group ( n=42 ) .The rats in SNx group were subjected to SNx.Sham surgery was performed in the rats in control group.The rats in SNx group were divided into subgroups at 0, 4, 8 and 12 weeks ( n=6) and the other rats in SNx group were divided into SNx+vehi-cle group, SNx+necrostatin-1 (Nec-1) group and SNx+3-methyladenine (3-MA) group.The expression of RIP1, RIP3, LC3 and beclin-1 at mRNA and protein levels was measured at 0, 4, 8 and 12 weeks by qPCR and immunohistochemistry. The effects of Nec-1 or 3-MA on the protein expression of LC3-I, LC3-II and beclin-1, and production of reactive oxygen species ( ROS) in the rat kidney were determined by Western blot and DCFH-DA staining.The death of renal tubular epi-thelial cells in the SNx rats was observed by TUNEL staining and electron microscopy.Finally, the effects of Nec-1 and 3-MA on blood urea nitrogen ( BUN) , serum creatinine ( SCr) and the pathological changes of the renal tissues were ana-lyzed.RESULTS:The highest mRNA and protein levels of RIP1, RIP3, LC3 and beclin-1 appeared at the 8th week after SNx (P<0.01).Compared with the rats in SNx+vehicle group, the protein over-expression of LC3-II/I and beclin-1, re-nal tubular epithelial cells with typical morphological features of necroptotic cell death and TUNEL-positive renal tubular
cells were decreased in the SNx rats treated with Nec-1 and 3-MA (P<0.01), but 3-MA did not reduce the increased con-centration of ROS.In addition, treatment with Nec-1 and 3-MA obviously reduced BUN, SCr (P<0.05), glomeruloscle-rosis index and tubulointerstitial injury score (P<0.01).CONCLUSION:Autophagy participates in the excessive death of renal tubular epithelial cells in SNx rats.Inhibition of autograph prevents necroptotic cell death of renal tubular cells, and alleviates chronic renal injury in SNx rats.

Objective To have a knowledge of the mutagenicity of contaminates in drinking water in a certain city. Methods Mutagenic activity of organic extracts of source water chloridized water in water plant and tap water from the city were detected with single cell gel-electrophoresis mouse primary hepatocytes in culture were used and Ames test. Results For Ames test the organic extract in 3 liters chloridized water in water plant and tap water samples produced positive result and that in 6 liters source water sample still gave a negative result. For single cell gel-electrophoresis test the organic extract in 0.1 liter chloridized water in water plant tap water and 0.5 liter source water produced positive result. Conclusion The sensitivity of single cell gel-electrophoresis with primary hepatocytes in detection of genotoxicity of cloridized water and source water is much higher than Ames test.

Objective To establish the data including anatomy and histology of main organs in Rongshui miniature pig (RMP).Methods F1 Rongshui miniature pigs with male and female (2 in each group) in 6 month old were used in this experiment.We measured body weights, dissected these pigs after anaesthesia, recorded total blood volume, total plasma volume, number of spine and dental formula, took main organs for photographs, and made histological sections observed and took photographs by microscope.Results We gained the photographs of main organs and histological sections, organ weights,organic coefficients and other basic data.Conclusion Basic anatomy and histology data of main organs in RMP were collected.

Objective To explore a better method for treatment atrophic rhinitis.Methods 56 patients with atrophic rhinitis(96 lateral)were treated by nasal submucou pediculated bone-suberiosteal muscle flap extracted from anterior wall of sinus maxillaries.Results All patients were followed 2 to 10 years,total effective rate was 100 %, with 49 cases(87.5 %)showing prominent effect.Conclusion The grafted flap cannot be assimilated,felled off and necrosis,because the flap has rich blood supply.This methods has obvious short-term effective and stable long-term effective.No complications were found.

OBJECTIVETo investigate the antioxidant effect of melatonin (MT) in the rats with phosgene-induced lung injury and its possible mechanism.

METHODSFifty male SD rats were equally randomized into phosgene exposure group, air control group, MT treatment group, dexamethasone (DX) treatment group, and negative control group. All groups except the air control group were exposed to 8.33 mg/L phosgene for 5 min, and the MT treatment group, DX treatment group, and negative control group were injected with MT (10 mg/kg), DX (2.5 mg/kg), and 1% ethanol saline (1 ml/kg), respectively, via the caudal vein 1 hour after exposure. The rats were sacrificed 6h later. Then, the wet/dry ratio of the lung, the total protein content and neutrophil count in bronchoalveolar lavage fluid (BALF), and the malonaldehyde (MDA) content and superoxide dismutase (SOD) and myeloperoxidase (MPO) activities in lung homogenate were measured; pathological observation was made on the lung tissue under an optical microscope; the protein expression of inducible nitric oxide synthase (iNOS) and NF-κB in the lung tissue was measured by Western blot.

RESULTSCompared with the air control group, the phosgene exposure group showed significantly increased wet/dry ratio of the lung and total protein content and neutrophil count in BALF (P < 0.01) as well as significantly increased MDA content and MPO activity in the lung tissue (P < 0.05). Compared with the phosgene exposure group, the MT treatment group showed significantly decreased MDA content and MPO activity and significantly increased SOD activity (P < 0.01), and the MT treatment group and DX treatment group showed significantly decreased protein expression of iNOS and NF-κB (P < 0.01).

CONCLUSIONMT has protective effect in phosgene-induced lung injury, and its protective mechanism may be associated with scavenging free radicals and inhibiting expression of iNOS and NF-κB.

Acute Lung Injury ; chemically induced ; metabolism ; prevention & control ; Animals ; Disease Models, Animal ; Male ; Malondialdehyde ; metabolism ; Melatonin ; pharmacology ; therapeutic use ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Peroxidase ; metabolism ; Phosgene ; toxicity ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the expression of lipopolysaccharide (LPS) receptor CD14 and Toll-like receptor 4 (TLR4) on Kupffer cells and its role in ischemia-reperfusion injury (IRI) on rat liver graft.

METHODSThe Kupffer cells following IRI were isolated and divided into control, ischemia-reperfusion (IR), and anti-CD14 antibody group. The mRNA and protein expression of CD14 and TLR4, nuclear factor kappa B (NF-kappaB) activity and TNF-alpha level in supernatant were measured.

RESULTSThe mRNA and protein expression of CD14 and TLR4 in IR group were significantly higher than those in control group (P < 0.01). The NF-kappaB activity and TNF-alpha level in IR group were significantly higher than those in control group (P < 0.01), and they greatly decreased after anti CD14 antibody treatment (compared with IR group, P < 0.05), but were still significantly higher than those in control group (P < 0.01).

CONCLUSIONSLPS following IRI could up-regulate CD14 and TLR4 gene and protein expression on Kupffer cells, and subsequently activate NF-kappaB to produce cytokines, but other signal transduction pathways might also participate in the IRI.

Animals ; In Vitro Techniques ; Kupffer Cells ; pathology ; physiology ; Lipopolysaccharide Receptors ; genetics ; metabolism ; Liver Transplantation ; pathology ; physiology ; Male ; NF-kappa B ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; pathology ; physiopathology ; Signal Transduction ; Toll-Like Receptor 4 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; secretion

Objective To quantitative the changing information of estrogen receptor βgene which was in tissue and organ of sex gland during oestrus and dioestrus of Beagles, and to show the different expression situation of hypothalamus-pituitary-gonad axis during oestrus and dioestrus, and providing the basic of theory to research deeply the mechanism of heat of Beagles. Methods As the key gene in regulation reproduction, ERβgene is located in hypothalamus-pituitary-gonad axis, so using Beagles which was in oestrus and dioestrus, and extract the RNA from hypothalamus、pituitary、ovary and uterus respectively,after reverse transcription we detected the expression of ERβgene by real-time quantitative PCR.Results The expression of ERβgene mRNA from ovary、uterus、pituitary、hypothalamus of Beagles which was in dioestrus was 0.35 times, 0.17 times, 0.44 times and 0.43 times than the expression of ERβgene mRNA from ovary, uterus, pituitary, hypothalamus of Beagles which was in oestrus.Conclusion The expression of ERβgene was up-regulation in hypothalamus-pituitary-ovary axis of Beagles which was in oestrus.

OBJECTIVETo study the difference in the expression of VEGF, bFGF and their receptors between young and postmenopausal women with breast cancer.

METHODSThe expression of VEGF, FLK-1, bFGF and FLG in 40 young and 30 postmenopausal women with breast cancer was studied by immunohistochemical method (SABC), with its relation with axillary lymph node metastasis and the clinical and pathologic characteristics. The expression index between these two groups was compared.

RESULTSThe positive axillary lymph node rate and the mean expression of VEGF, bFGF in the young group were higher than postmenopausal group (P < 0.01 and P < 0.05), respectively. The mean expression of VEGF, bFGF, FLK-1 and FLG of axillary lymph node positive patients was higher than the negative ones both in young and postmenopausal women groups (P < 0.05 and P < 0.01). There was also a significant difference in VEGF, bFGF, FLK-1, FLG and MVC between the stage 0 - II and stage III - IV (P < 0.05 and P < 0.01) in both groups.

CONCLUSIONBreast cancer angiogenesis, characterized by the high expression of VEGF and bFGF, is directly correlated with the high tumor aggressiveness in the young women.

Adult ; Age Factors ; Aged ; Breast Neoplasms ; chemistry ; pathology ; Female ; Fibroblast Growth Factor 2 ; analysis ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Middle Aged ; Postmenopause ; Receptor, Fibroblast Growth Factor, Type 1 ; analysis ; Receptors, Estrogen ; analysis ; Vascular Endothelial Growth Factor A ; analysis ; Vascular Endothelial Growth Factor Receptor-2 ; analysis

Objective:To evaluate the efficacy and safety of thalidomide on preventing and treating radiation-induced oral mucositis (RIOM) in nasopharyngeal carcinoma (NPC). Methods:A total of 60 patients with NPC were randomly divided into treatment group and control group. The treatment group took thalidomide and gargled with mixture of saline+gentamycin+dexamethasone when ra-diotherapy started, and the control group gargled only with mixture of saline+gentamycin+dexamethasone. The time of occurrence and degree of RIOM in both groups were registered at the same time. Results:The incidence of RIOM in treatment group was lower than that in control group (P<0.05). For the incidence of 3 or 4 grade RIOM, the treatment group was also lower than the control group. No statistical difference in terms of objective response rate was found between the two groups (P>0.05). There was no statisti-cally significant difference between the treatment group and the control group in the oral average dose, V30, V35, V40, V45, V50, and V54 (P>0.05). There was no statistically significant difference between the treatment group and the control group in the use of nutri-tion agents, hormones and antibiotics (P>0.05). There was no statistical difference between the groups in the drowsiness, peripheral nerve toxicity, hematologic toxicity and gastrointestinal reaction (P>0.05). Conclusion:Thalidomide can reduce the incidence and se-verity of RIOM for radiotherapy of patients with NPC, which deserves clinic application.