Aim To investigate the molecular mechanisms of differentiation in human leukemia HL-60 cells induced by diallyl disulfide(DADS)using suppression subtractive hybridization(SSH).Methods In our privious study,the subtractive cDNA library was constructed successfully and efficiently. 30 clones were randomly analyed with restriction enzyme.The inserts of cDNAs were analyzed by restrictive enzyme EcoR I.Positive clones were sequenced and the homology of resulting cDNA sequences were analyzed through bioinformatics software Blastn.Results 18 clones contained 100～600 bp cDNA inserts.10 differantiation genes were obtained and involved in cell cycle,signal transduction,metabolism and RNA binding.And 3 of 10 genes,p21,STAT1 and CAMTA1 were up-regulated and detected by RT-PCR,the results matched with SSH.Conclusion sThere are tight correlation between the differentiation induced by DADS and three-upregulated gene:p21,STAT1 and CAMTA1.

Air Pollution ; prevention & control ; Epidemiologic Research Design

OBJECTIVETo analyze the causes of death and potential years of life lost (PYLL) in 7 districts of Guangzhou from 2003 to 2005.

METHODSThe bureau of Health of Guangzhou provided the data of 70 201 cases of death occurring between 2003 and 2005. The top 10 causes of death were analyzed and the PYLL and crude death rate were calculated using mortality difference dissembling method.

RESULTSBetween 2003 and 2005, the top 3 causes of death in Guangzhou were diseases of the circulatory system, neoplasms and respiratory diseases. Aging of the population led to an increased death rate. The major causes of death affecting PYLL consisted of neoplasms, diseases of the circulatory system, injuries and poisoning.

CONCLUSIONPriority should be given to the control of diseases of the circulatory system in the health care in Guangzhou. Neoplasms, injuries and poisoning all contribute to a high PYLL.

Objective To assess the application of three-dimensional digital subtraction angiography (3D-DSA) in evaluation of ruptured intracranial aneurysm after clipping and to discuss the different variable use of vol-ume rendering(VR), gradient rendering (GR) and maximum intensity projection (MIP). Methods From January 2011 to December 2012 , 88 patients with 92 ruptured intracranial aneurysms were treated with clipping using titani-um clips in our hospital and followed up by both 2D-DSA and 3D-DSA. Residual aneurysms , Clips place, clips and parent arteries and stenosis of parent arteries were evaluated by volume rendering (VR), gradient rendering (GR) and maximum intensity projection (MIP). Results Among 92 clipped aneurysms, 23 residual aneurysms were found by 3D-DSA. Residual aneurysms were recorded according to the Sindou grade: 15 of gradeⅠ, 3 of gradeⅡ, 4 of grade Ⅲand 1 of grade Ⅳ. Three patients of grade Ⅲand 1 of grade Ⅳwith residual aneurysms were retreated by clipping or coiling, and 1 patient of grade Ⅲ was dead with rupture of residual aneurysm. The clips and number of clips were clearly visualized , and relationship between the clips and the aneurysms was well demonstrated by VR, GR and MIP images. VR, GR images showed the remnants clearly. Three-dimensional digital subtraction angiography did not showed accurate details of the stenosis of parent arties which required an analysis of 2D-DSA. Conclusion Three-dimensional digital subtraction angiography can be used for definite evaluation of resid-ual aneurysms after clipping, especially by VR, GR images. It is helpful to manage the residual ruptured aneurysms.

AIM: To study the influence of injection agent of pidotimod on function of the immune-system in normal and immune-depressed mice, and to investigate the mechanisms of the immunomodulating action of pidotimod. METHODS: The expressions of TNF-? and IL-6 in mice spleen was detected by RT-PCR method. The effects of pidotimod on phagocytosis functions of mice peritoneal exudate cells was investigated by neutral red phagocytosis. The lymphocyte proliferations induced by Con A or LPS were detected by MTT method. Adopt the serum haemolysis to measure the production of serum antibody. RESULTS: The normal and immune-depressed mice were treated for 14 days with different dosages of pidotimod by injection. Pidotimod can significantly increase the expressions of TNF-? and IL-6 of spleen, potentiate phagocytosis of the peritoneal exudate cells, potentiate the lymphocyte proliferations ability induced by Con A or LPS. CONCLUSION: Pidotimod can potentiate amelioration normal and immune-depressed mice immunesystem function and increase expressions of TNF-? and IL-6 of spleen.

In the current study, nine flavonoids were isolated and purified from 75% ethanol extract of Selaginella uncinata (Desv.) Spring by column chromatographic techniques over macroporous resin, polyamide, silica gel, Sephadex LH-20 and pre-HPLC. On the basis of their physico-chemical properties and spectroscopic data analyses, these compounds were elucidated as cirsimarin (1), nepitrin (2), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (3), apigenin-6-C-β-D-glucopyranosyl-8-C-α-L-arabinopyranoside (4), apigenin-7-O-β-D-glucopyranoside (5), 2,3-dihydroamentoflavone (6), 4'-O-methylamentoflavone (7), 2,3-dihydro-4'-O-methyl-amentoflavone (8), and 2,3,2",3"-tetrahydron-4'-O-methyl-robustaflavone (9). Compounds 1-5 belong to flavonoid glycosides and were isolated from the genus Selaginella for the first time.
Flavonoids ; analysis ; Selaginellaceae ; chemistry

Aim Toresearchthemolecularmecha-nisms of DADS-induced apoptosis in human leukemia K562cells.Methods Cellviabilitywasmeasuredby MTT. Levels of DADS-induced ROS were measured by 2ˊ, 7ˊ-dichlorofluorescein diacetate ( DCFH-DA) fluo-rescence. DADS-induced mRNA levels of components of the NADPH oxidase were detected by Real-time PCR. The combination of protein Rac2 and p67phox was measured by immunoprecipitation assays. Flow cy-tometry methods were used to determine the percentage of apoptosis cells. DADS-induced Rac2 levels were measuredbyWesternblot.Results TheDADS-trea-ted K562 cells showed a dose-and time-dependent de-crease in cell viability and proliferation. There was sig-nificant up-regulation of the mRNA level of components of the NADPH oxidase complex in K562 cells after treatment with 6 mg·L-1 DADS for 6 h. Western blot results revealed that, compared with the control group, there was a significant up-regulation of Rac2 protein in K562 cells treated with 5. 0 and 10. 0 mg·L-1 DADS for 24h. And Rac2 combined with p67phox in DADS-induced apoptosis in K562 cells. PMA markedly in-creased the percentage of apoptotic cells, and DPI re-duced the percentage of apoptotic cells in DADS-in-duced K562 cells. Levels of DADS-induced ROS, also showed enhancement when exposed in PMA, but there was no DADS-induced ROS production evident when exposed in DPI in DADS induced K562 cells. Conclu-sions TheseresultsindicatethatNADPHoxidaseis the main source of DADS-induced ROS production. Diallyl disulfide induces apoptosis in human leukemia K562 cells through activation of NADPH oxidase.

AIM: To discuss the clinical efficacy of fresh amniotic membrane ( AM ) during the microscopic adjustable suture surgery in children's intercommunity strabismus, in order to guide clinical treatment.
METHODS: With the clinical randomized control study (RCT), 60 (112 eyes) cases of patients in childhood who received microscopic strabismus surgery in our hospital were divided them into two different groups from Jan. 2010 to Oct. 2015. According to the application of AM on the basis of ophthalmology outpatient number, 30 cases (58 eyes) in group A were treated with rectus muscle recession surgery combined adjustable suture combined with AM. The other 30 cases (54 eyes) in group B were treated with rectus muscle recession surgery combined adjustable suture only. All patients in two groups were followed-up over 6mo after the strabismus surgery.
RESULTS:Twenty-seven cases ( 48 eyes ) of all the strabismus patients must be adjusted after strabismus surgery, and the eye position adjustment rate was 42.9%. At 1mo after surgery, eye position of 18 cases (29 eyes) can be adjusted in all patients, and 44. 8% (16 cases, 26 eyes ) in group A with the average of adjustment lengths was 2. 56±0. 64mm, and 5. 6% ( 2 cases, 3 eyes ) in group B, with the average of adjustment lengths was 0. 52±0. 28mm, the differences of the adjustment rate and the average of adjustment amount were both high statistically significant (χ2 =22.477, P<0. 01; t=16. 502, P<0. 01 ) between the two groups. Except of 3 cases who couldn't cooperate with eye position adjustment, they all received eye position adjustment in different degrees in one month after strabismus surgery,and after eye position adjustment, 27 cases (53 eyes) in group A got normal eye position, and the correction rate of eye position was 91. 4%, and 16 cases (28 eyes) in group B got normal eye position after eye position adjustment, the correction rate was 51. 9%, the differences of the correction rate were statistically significant (χ2=21. 827, P<0. 01) between the two groups.
CONCLUSION: The application of fresh AM in the microscopic adjustable suture strabismus surgery is exactly effective in treatment of children's intercommunity strabismus. It can significantly extend the adjustment time and increase the adjustment amount, and it also can statistically improve the controllability and achievement ratio for children's strabismus surgery.

Aim To study the effects of cycle arrest and molecular mechanism in human leukemia HL-60 cells induced by diallyl disulfide ( DADS ) . Methods Cell count, colony formation in soft agar experiments and flow cytometry analysis were employed to observe the DADS-induced cell growth inhibition and the effect of cycle arrest in HL-60 cells. The expressions of Chk1/2 and its downstream element in HL-60 cells were detected by Western blot. Results Cell count revealed that population doubling time increased to 35. 03 h and 71. 82 h, respectively, from 19. 14 h in HL-60 cells treated with 60 and 120 μmol·L-1 DADS ( P<0. 05 ) . Colony formation in soft agar experiments showed that colony formation inhibition rate of HL-60 cells exposed to 30, 60, 90 and 120μmol·L-1 DADS increased to 35. 06%, 62. 10%, 93. 79% and 99. 35%, respectively ( P<0. 05 ) . Flow cytometry a-nalysis exhibited that HL-60 cells treated with 60 and 120 μmol · L-1 DADS for 24 h and 48 h arrested in G2/M phase in a concentration-and time-dependent manner ( P <0. 05 ) . Western blot disclosed that the expression of p-Chk1 increased in a time-dependent manner ( P <0. 05 ); however, Chk1, Chk2 and p-Chk2 were not changed in HL-60 cells treated with 60μmol·L-1 DADS (P >0. 05). The expression of Cdc25C, CyclinB1 and CDK1 decreased after treated with 60 μmol·L-1 DADS in a time-dependent manner ( P<0. 05 ) , but the expression of 14-3-3 protein did not change ( P>0. 05 ) . Conclusion DADS can in-hibit the proliferation of HL-60 cells, and induce G2/M arrest through Chk1/Cdc25 C/CyclinB1/CDK1 path-way.

Objective - （ ） To investigate the effects of nuclear factor erythroid 2 related factor 2 NRF2 on the oxidative stress （ ） Methods ） ，， induced by trichloromethane TCM in human normal hepatocyte L02 cells. i L02 cells were stimulated with 1 2 ， ， ， （ ）， 4 8 12 16 and 20 mmol/L TCM solution dissolved in dimethyl sulfoxide and the control group and blank group were set ， - ， up. After culturing for 24 hours the cell viability was detected by CCK 8 colorimetric method and the concentration of TCM ） -， - stimulation was screened. ii L02 cells in logarithmic growth phase were randomly divided into control group and low medium - ， ， ， and high dose groups. After 24 hours of exposure to 0 4 8 and 12 mmol/L TCM the cells were collected. The activity of （ ）， （ ）， （ - ） （ ） superoxide dismutase SOD catalase CAT glutathione peroxidase GSH Px and the level of malondialdehyde MDA NRF2， - （HO-1）， were detected by colorimetric analysis. The mRNA expression levels of heme oxygenase 1 glutamate cysteine （GCLC） （） （NQO1） - ligase catalytic subunit and NAD P H quinone dehydrogenase 1 were detected by real time fluorescence ， - ， polymerase chain reaction. The protein levels of NRF2 HO 1 GCLC and NQO1 were detected by Western blotting.Results ） ， ， ， ， i When the concentration of TCM was 4 8 12 16 and 20 mmol/L the survival rate of L02 cells decreased （ P ） ， ， significantly compared with the control group all <0.05 . The concentration of 0 4 8 and 12 mmol/L were selected as the ） ， - stimulation doses for subsequent experiments. ii Compared with the control group the activities of SOD and GSH Px in L02 （ P ） （ P ）， - cells in the three doses groups decreased all <0.05 and the levels of MAD increased all <0.05 with a dose effect - （P ）， relationship. The CAT activity of L02 cells in the medium dose group was lower than that in the control group <0.05 and the - （ P ） CAT activity of L02 cells in the high dose group was lower than that in the others three groups all <0.05 . Compared with the ， NRF2 - （P ），NRF2 control group the relative expression levels of mRNA in L02 cells in the low dose group decreased <0.05 - （P ）， NRF2 mRNA in L02 cells in the medium dose group increased <0.05 mRNA and NRF2 protein expression in L02 cells in （ P ） HO-1，GCLC， NQO1 ， the highdose group increased both <0.05 . The relative expression level of mRNA and GCLC NQO1 （ P ） protein expression in L02 cells in the three doses groups increased compared with the control group all <0.05 . The relative NRF2 - - - expression level of mRNA in L02 cells in the high dose group was higher than that in the low and medium dose groups （ P ）， - （P ）， both <0.05 and the relative expression of NRF2 protein was higher than that in the low dose group <0.05 but the HO-1 GCLC - - （ relative expression levels of and mRNA and HO 1 protein level were lower than those in the medium dose group all P ）Conclusion - <0.05 . TCM exposure can inhibit the proliferation of L02 cells by inducing oxidative stress with a dose effect ， relationship. In this process the antioxidant mechanism mediated by NRF2 was activated. The expression of antioxidant defense ， - ， and detoxification related target genes downstream of NRF2 signaling pathway was activated and the expression of HO 1 - GCLC and NQO1 was up regulated to alleviate the oxidative damage caused by TCM.