Selective protein degradation by the ubiquitin 26S proteasome pathway has emerged as a key regulatory mechanism in a wide variety of cellular processes.The ubiquitin/26S proteosome pathway mainly consists of ubiquitin activating enzyme(E1),ubiquitin conjugating enzyme(E2),ubiquitin protein ligase(E3),and 26S proteasome.In an ATP-dependent reaction,uibquitin(Ub) is conjugated to E1,the activated Ub is then transferred to an E2.Finally,the Ub-E2 intermediate delivers the Ub to the target protein by E3 recognition.Polyubiquinated proteins are eventually degraded by the 26S proteasome.In plants,regulated protein degradation by /26S proteasome pathway contributes significantly to development by affecting a wide range of progress,including hormone signaling,photomorphogenesis,self-incompatibility and cell cycle.The recent progress towards understanding the role of the Ub/26S proteasome pathway during plant development was reviewed.

Objective To investigate the polymorphism of glutathione S-transferease P1 gene(GSTP1) and the association between the mutation and susceptibility in childhood asthma.Methods The distribute frequency of Ile105/Ile105,Ile105/Val105 and Val105/Val105 ge-notype in GSTP1 of 51 children with asthmatic and 40 normal children were studied with polymerase chain reaction-restriction tragment length polymorphism(PCR-RFLP).Results The frequencies of Ile/Ile Ile/Val,Val/Val genotype were 66.7%,27.4% and 5.9%,the frequencies of Ile,Val allele were 80.4% and 19.6% in the asthmatic group.But the frequencies of Ile/Ile,Ile/Val,Val/Val genotype were 90.0%,7.5% and 2.5%,the frequencies of Ile,Val allele were 93.8%,6.2% in control group.The frequencies Ile/Val,Val/Val genotype and Val allele in asthmatic group were more than that in control group.A significant difference was found in the frequency distribution of GSTP1 genotypes between two groups(?2=6.947 P

Objective To understand the pathogen distribution situation among the children inpatients with hand ,foot and mouth diseases(HFMD) in Guiyang area during 2012 to provide the basis for the diagnosis ,treatment and prevention .Methods The data in 3 179 cases of HFMD were collected .The fluorescence quantitative RT‐PCR was adopted to perform the genotyping on universal enterovirus ,enterovirus 71(EV71)and Coxsackie virus A16(CA16) .Results A total 3 179 samples of HFMD were col‐lected ,among them ,151 cases (4 .75% ) were CA16 positive ,331 cases (10 .41% ) were EV71 positive ,7 cases (0 .22% ) were CA16 and EV71 co‐infection ,and 897 cases(28 .22% )were the other enterovirus .The whole year had 2 peaks of onset ,which were April to July and October to November .The onset age focused on the children aged under 5 years old (96 .16% ) ,among them ,0-3 years old had the highest onset ,moreover male children were more than female .Conclusion The etiology distribution of children HFMD in Guiyang area during 201 was dominated by the other genotypes of enterovirus and EV71 .

Objective To study the distribution of genotypes of hepatitis C virus in Guizhou and its relationship between infec-tious route of genotype and age ,gender was analyzed .Methods Serum specimens in this study were obtained from 198 patients , whose anti-HCV and HCV RNA were positive .A reverse transcriptase PCR(RT nested-PCR)assay using conserved primers de-duced from the core-envelope 1(C-E1)region of the hepatitis C virus(HCV)genome was employed to amplify a 474-nucleotide-long fragment .Phylogenetic analysis of the C-E1 sequences was conducted by direct sequencing of the RT-PCR products and alignment with published HCV subtypes in GenBank .Subtypes of the samples were determined by nucleotide sequencing followed by composi-tion of a phylogenetic tree .Results Among the 198 patients surveyed ,genotype 1a was detected in 4 cases(2 .0% ) ,genotype 1b in 71 cases(35 .9% ) ,genotype 2a in 9 cases(4 .6% ) ,genotype 3a in 29 cases(14 .7% ) ,genotype 3b in 47 cases(23 .7% ) ,genotype 6 a in 37 cases(18 .7% )and genotype 6d in 1 cases(0 .5% ) .Genotype distribution on gender had no statistical significance(P>0 .05) , and its distribution on people with different ages had statistical significance(P<0 .05) ,and its distribution on patients with different infectious routes was significantly different(P<0 .05) .Conclusion The major genotypes of HCV are 1b ,3b ,6a and 3a in Guizhou , and genotype 1a is predominant .Genotypes 1a ,2a and 6d exist too .Genotypes of patients infected with HCV are related to their in-fectious routes ,and the HCV genotypes are in a great variety .

OBJECTIVETo study genetic damage of mice caused by the volatile organic compounds (VOC) of decoration materials.

METHODSFifty-five hotel guest rooms newly decorated within 6 months and 18 hotel guest rooms not decorated within 3 years were selected to determine the concentrations of 6 main VOC (benzene, methylbenzene, dimethylbenzene, ethyl acetate, butyl acetate, formaldehyde) in the air. Mice were exposed to VOC with the concentrations of 5, 10, 20, 40 times respectively as high as those present in the newly decorated rooms in an exposure cabinet for 15 days. DNA damage of peripheral lymphocytes of the mice was determined by single cell gel electrophoresis (SCGE) and bone marrow micronucleus test.

RESULTSThe concentrations of benzene, methylbenzene, dimethylbenzene, ethyl acetate, butyl acetate and formaldehyde in the rooms newly decorated within 6 months (6.50, 3.00, 6.70, 41.33, 1.70 and 0.14 mg/m(3) respectively) were significantly higher than those in rooms not decorated within 3 years (0.08, 0.94, 1.38, 0.25, 0.25, 0.01 mg/m(3), P < 0.01). DNA damage rates of peripheral lymphocytes in the concentrations of 10, 20, 40 times of exposure groups were significantly higher than those in the control groups (P < 0.05 or P < 0.01), and the frequencies of micronucleus in the mice exposed to 40 times of concentration was significantly higher than that in control group.

CONCLUSIONHigh concentrations of the volatile organic chemical compounds may cause genetic damage in mice. SCGE test is more sensitive than micronucleus test.

Air Pollutants ; toxicity ; Air Pollution, Indoor ; adverse effects ; Animals ; DNA Damage ; drug effects ; Mice ; Micronucleus Tests ; No-Observed-Adverse-Effect Level ; Organic Chemicals ; toxicity ; Paint ; adverse effects

OBJECTIVETo compare the effect of laparoscopic radical operation and open operation on systemic immunity in patients with colorectal cancer.

METHODSSixty patients with colorectal cancer were randomly divided into laparoscopic and open operation groups from March 2004 to December 2004, each group had 30 cases. CRP, IgA, IgM, IgG, CD3 (+) cells, CD4 (+) cells, CD8 (+) cells, NK cells, CD4 (+) CD5RA (+) cells, CD4 (+) CD45RO (+) cells and lymphocytes in peripheral blood were counted and compared on the 1st day before operation, 3rd and 7th day after operation.

RESULTSThe two groups were comparable as for age, tumor location and stages. In open operation group, lymphocyte counts were (1.09+/- 0.29) x 10(9)/L, CD4 (+) cell (0.54 +/- 0.14) x 10(9)/L, CD8 (+) cell (0.31 +/- 0.08) x 10(9)/L, CD4 (+) CD45RO (+) (61.1+/- 8.9)%, and IgM level (136.9+/- 52.8) IU/ml, IgG (115.2 +/- 45.7) IU/ml on the 3rd day after operation, CD8 (+) cell counts were (0.32 +/- 0.09) x 10 (9)/L, CD4 (+) CD45RO (+) cell (63.2 +/- 9.1)% on the 7th day after operation, were all significantly lower than those on the 1st day before operation respectively(P< 0.05, P< 0.01). In laparoscopic operation group, the decreases of such parameters except CD4 (+) CD45RO (+) cell (62.7 +/- 12.5)% were not obvious on the 3rd day after operation. There were significant difference in lymphocyte counts (1.29 +/- 0.37) x 10( 9 )/L, IgM (164.5 +/- 48.2) IU/ml and CD8 (+) cell counts (0.38 +/- 0.09) x 10 (9) /L on the 3rd day after operation between two groups (P< 0.05).

CONCLUSIONCompared with open radical operation, laparoscopic radical operation has predominance in protecting systemic immunity to treat colorectal carcinoma.

Aged ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; immunology ; Colorectal Neoplasms ; immunology ; surgery ; Female ; Humans ; Killer Cells, Natural ; immunology ; Laparoscopy ; Lymphocyte Count ; Male ; Middle Aged

OBJECTIVETo investigate the effects of hydroquinone (HQ) on expression of ubiquitin-ligating enzyme Rad18 in human hepatic cells (L-02), and to explore the role and possible mechanism of Rad18 involved in toxicity of HQ to hepatic cells.

METHODSAfter L-02 hepatic cells were exposed to HQ with various concentrations (0, 5, 10, 20, 40, 80 and 160 micromol/L) for 24 h, cell survival rate was measured by MTT assay; DNA impairment was evaluated by single cell gel electrophoresis (SCGE); The expression levels of Rad18 mRNA and protein were detected by Real-time fluorescent quantitative polymerase chain reaction (QPCR) technique and Western blot method respectively.

RESULTSHQ with concentration from 0 to 80 micromol/L had little effect on survival rate of L-02 (P > 0.05); Whereas the survival rate in the group of 160 micromol/L was significantly lower than in the control with the significant difference (P < 0.01) after treated with HQ for 24 h; The higher dose of HQ presented, the more degrees of olive tail moment (OTM) were produced and a dose-dependent relationship was shown. HQ in a low concentration (0 to approximately 40 micromol/L) could induce increase in the expression of Rad18 mRNA and protein which was in proportion to the increment of HQ concentration; the expression of Rad18 mRNA was enhanced increasingly, while the expression of Rad18 protein unchanged basically once the concentration of HQ exceeded 40 micromol/L; Besides, there was a positive correlation between OTM and the expression level of Rad18 mRNA (r = 0.919, P < 0.01).

CONCLUSIONHQ could regulate up the expression of Rad18 in L-02 hepatic cells.

Cell Survival ; drug effects ; Cells, Cultured ; DNA Damage ; drug effects ; DNA-Binding Proteins ; metabolism ; Hepatocytes ; drug effects ; enzymology ; Humans ; Hydroquinones ; toxicity ; Ubiquitin-Protein Ligases

OBJECTIVETo investigate the effects of hydroquinone (HQ) on expression of Polymerase eta (Pol eta) and DNA damage in human hepatic cells (L-02), and to explore the role and possible mechanism of Pol eta involved in the process of DNA damage-tolerance.

METHODSAfter L-02 hepatic cells were exposed to HQ with various concentrations (0, 5, 10, 20, 40, 80 and 160 micromol/L) for 24 h, cell survival rate was detected by MTT assay; DNA impairment was detected by single cell gel electrophoresis (SCGE); Real-time fluorescent quantitative PCR and Western blotting methods were used to measure the expression of Pol eta at the mRNA and protein level in L-02 hepatic cells exposed to HQ with various concentrations (0, 5, 10, 20, 40, 80 and 160 micromol/L).

RESULTSMTT assay showed that HQ with concentrations from 0 to 80 micromol/L had little effect on the survival rate of L-02 (P>0.05); whereas the survival rate of the group of 160 micromol/Lwas significantly higher than that of the control (P<0.01) after being treated with HQ for 24 h; the higher dose of HQ presented, the more degrees of DNA damage were produced. It was found that HQ in a low concentration (1-80 micromol/L) could induce the expression of Pol eta which was in proportion to the increasements of HQ concentration; the expression levels of mRNA and protein were reached to the maximum when treated with 80 micromol/L; the expression of Pol eta decreased (the relative quantity values were 2.32 +/- 0.16 and 1.20 respectively) once the concentration of HQ exceeded 160 micromol/L as compared with the group of 80 micromol/L, but it was higher than that of the control.

CONCLUSIONThis study suggested that Pol eta might involve in the process of DNA damage-tolerance induced by HQ in the hepatic cells.

Cell Survival ; drug effects ; Cells, Cultured ; DNA Damage ; drug effects ; DNA Repair ; DNA-Directed DNA Polymerase ; metabolism ; Hepatocytes ; drug effects ; metabolism ; Humans ; Hydroquinones ; adverse effects ; Mutagens

OBJECTIVETo investigate the effect and mechanism of FTY720 on acute graft versus host disease (GVHD) in rat small bowel transplantation (SBTx).

METHODSHeterotopic SBTx was performed using a parent (WF)-into-F1 (WFxACI) rat combination. Recipient rats were divided into experimental group (n=6) and control group (n=6). Rats in the experimental group were administered with FTY720 at 0.5 mg/kg for 14 days. Lymphocyte apoptosis in the liver and the mucosa of intestine and graft was detected by TUNEL and flow cytometry 15 days after transplantation. Recipient survival and lymphocyte apoptosis were compared between the two groups.

RESULTSRecipients in the control group died of GVHD after a mean survival time of (16+/-2.1) days. FTY720-treated recipients had a significantly longer survival (>100 days). After administration of FTY720, the percentage of apoptotic lymphocytes was significantly increased in the graft as compared to that in the control group by flow cytometry. The ratio of apoptotic lymphocyte in the liver and graft was also significantly higher in the experimental group by TUNEL.

CONCLUSIONFTY720 effectively induces the lymphocyte apoptosis, inhibits the lesion of target tissues by GVHD, and prolongs the recipient survival.

Animals ; Apoptosis ; drug effects ; Fingolimod Hydrochloride ; Graft vs Host Disease ; immunology ; prevention & control ; Immunosuppressive Agents ; pharmacology ; Intestine, Small ; transplantation ; Lymphocytes ; cytology ; drug effects ; Male ; Propylene Glycols ; pharmacology ; Rats ; Rats, Inbred WF ; Sphingosine ; analogs & derivatives ; pharmacology ; Transplantation, Heterotopic

OBJECTIVETo explore the differential proteomic expression in human liver cells L-02 induced by different dosages of trichloroethylene (TCE).

METHODSHuman liver cells L-02 were treated with different concentrations of TCE and the solvent control (dimethylsulfoxide). The total cellular proteins were separated using 2DE and visualized with silver staining after TCE treatment. The images were analyzed with Image Master 2D Platinum 5.0 analysis software. The differentially expressed protein spots were identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-TOF-MS).

RESULTSFifteen protein spots with significant difference were found, and went upward or downward or disappeared after the stimulation of TCE with different dosages, which indicated that TCE induced the change of the proteomic expression in the liver cells. The mass spectrum identification and the IPI human database retrieval were used for identifying 9 proteins related to the L-02 Liver cells induced by TCE.

CONCLUSIONThe result provides an insight to TCE-related molecular mechanism and which might be useful for further study of the TCE-associated proteins and molecular markers.

Cell Line ; Dose-Response Relationship, Drug ; Electrophoresis, Gel, Two-Dimensional ; Hepatocytes ; drug effects ; metabolism ; Humans ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Trichloroethylene ; toxicity