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.

Objective To evaluate the influence of different surface roughness on final surface texture of glazed ceramic. Methods Sixty porcelain discs were fabricated and randomly divided into five groups (n=12), which were sequentially grounded with water-cooled sandpapers of decreasing abrasiveness (220# , 400# , 600# , 800# and 1200# grit), and then all the specimens were glazed. Before and after glazing, the surface roughness (Ra) μm of the same specimens were evaluated quantitatively with profilometer and qualitatively using scanning electron microscopy (SEM). Results The smoothest surfaces were obtained with 1 200# sandpapers griting prior to overglazing (Ra =0. 19 μm),and 220# sandpapers griting prior to overglazing displayed the roughest surfaces (Ra =0. 55 μm) ; each group displayed significant difference before and after glazing (P < 0. 05). The images obtained through SEM were consistent with the profilometer readings. Conclusions The results suggest that decreasing the surface roughness of dental ceramic prior to glazing is of great importance in order to obtain the smoothest surface after glazing.

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 observe the incidence of ocular fundus disease in preschool children examined by non-mydriatic fundus camera and evaluate its effectiveness compared with direct inspection shadow mirror.
METHODS: Three thousand eight hundred and ninety-six preschool children from April 2012 to October 2013 were examined by Topcon TRC-NW300 color fluorescence fundus camera and direct inspection shadow mirror, and images were saved immediately.
RESULTS: Detection rate of non - mydriatic fundus photography was higher than that of direct inspection shadow mirror. In 3 896 cases, 41 eyes were detected abnormal fundus accounting for 1. 05%. The retinal myelinated nerve fibers, morning glory syndrome, retinitis pigmentosa, congenital retinoschisis were common, accounted for 24. 39%, 21. 95%, 14. 63%, 12-20% respectively. The children eye diseases were often accompanied by abnormal vision (68. 30%), ametropia (63. 41%), strabismus (19. 51%).
CONCLUSION:Non-mydriatic fundus photography is a mydriatic method without medicine, so it is easy for preschool children to accept. Image results could directly display the fundus lesions. It shows important significance in the screening for preschool children eye diseases.

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.

AIMTo explore the effects of heme- heme oxygenase-1 (HO-1)-carbon monoxide(CO)-cyclic GMP (cGMP)on aortic vascular reactivity in endotoxemic rats and its molecular mechanism.

METHODSBy using isolated vascular ring tension detecting technique, cumulative responses of thoracic aortic rings (TARs)to phenylephrine (PE) were measured at 6 h after lipopolysaccharide administration. Effects on contractile responses to PE were measured under which the TARs were incubated with hemin (He, donor of CO), zinc-protoporphyrin-IX(ZnPP-IX, selective inhibitor of HO-1), or methylene blue (MB, inhibitor of guanylyl cyclase), respectively. The content of CO and the activity of HO-1 were measured. The protein and the gene expression of HO-1 were examined by Western blot and RT-PCR.

RESULTSContractile responses of TARs to cumulative doses of PE were depressed by pretreated with LPS. The hyporesponsiveness was partly reversed by incubation with ZnPP-IX and was restored to normal by incubation with MB in endotoxemic rats. Incubation with He could contribute to the vascular hyporeactivity. The content of CO, the activity and the protein and the gene expression of HO-1 were significantly increased in aorta of endotoxemic rats.

CONCLUSIONLPS could induce the HO-1 mRNA and the protein expression, the activity of HO-1 increase in aorta, lead to active the pathway of heme-HO-1-CO-cGMP, which is one of the important mechanisms of the vascular hyporeactivity in endotoxemic rats.

Animals ; Aorta ; drug effects ; metabolism ; Carbon Monoxide ; metabolism ; Cyclic GMP ; metabolism ; Heme Oxygenase (Decyclizing) ; metabolism ; Lipopolysaccharides ; adverse effects ; Male ; Phenylephrine ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

·AIM: To investigate the ocular surface and refractive change after the surgery of levator resection and frontalis suspension. ·METHODS:One hundred and twenty-one patients (146 eyes) for corrective surgery of congenital ptosis were selected in our department from July 2014 to June 2016. According to the severity of congenital ptosis, all the children divided into mild group (47 eyes), moderate group (68 eyes) and severe group(31 eyes). All the children were divided into group of less than 5 years old (104 eyes) and group of more than or equal to 5 years old (42 eyes) according to the age. According to the surgical approach, all the children divided into group of levator resection (62 eyes) and group of frontalis suspension(84 eyes). The effects of the two surgeries on the ocular surface and refractive were compared. The relationship between age, severity of postoperative ptosis and postoperative ocular surface, refractive of children were analyzed. ·RESULTS: There were no significant differences in break-up time (BUT), keratometry (Km) and corneal astigmatism (Ast) in the levator resection group and frontalis suspension group (P > 0. 05). There was significant difference between the mild, moderate and severe group on the proportion of excellent corrected to uncorrected (Z=-2.936, P=0.003). The proportion of excellent correction in mild group was higher than that of moderate and severe group(93.6%, 83.8% and 67.7%). The indexes of BUT (F=9.793, P=0.041) and Km (F=11.657,P=0.037) in the three groups decreased with the increase of severity. While the Ast (F=28.417, P<0.01) showed an increasing trend, the difference was significant. In addition, the proportion of excellent correction in <5 year old group was significantly more than≥5 years old group(x2=4.082,P=0.043). The index of Km (t= 2.813, P= 0.006) was higher and Ast (t=-7.741, P<0.01) was lower in the <5 year old group. There was no significant difference in the index of BUT between the two groups after surgery (P>0.05). · CONCLUSION: The corneal refractive power and astigmatism can be improved similarly after treated with levator resection and frontalis suspension surgery. And there is some damage of corneal epithelial in the initial postoperation, but the function of ocular surface can return to normal after a period of time. The less severity of ptosis before surgery,the better improvement of ocular surface and refractive after sugery. There is no correlation between the age and postoperative ocular surface, but earlier surgical treatment may improve the children's refractive power and astigmatism and achieve greater benefits.

Objective To analyze the willingness and influencing factors for usage of pre-exposure prophylaxis(PrEP) among male students who have sex with men (MSM) in universities of Guangxi. Meth-ods 295 MSM students in universities were recruited by Snow-balling methods. Questionnaires were self-administered to collect social demographic information,AIDS related risky sexual behaviors and willingness for usage of PrEP. Logistic regression was employed for univariate and multivariate analysis. Results Of the 295 MSM,58(19.7%)had ever heard PrEP,265(89.8%) said that they would like to use PrEP. Multivari-ate logistic regression analysis showed that those who had found partners through friends ( OR=11.419,95%CI:1.363~95.641), those who would advise his friend to use PrEP ( OR=87.946,95%CI:13.660-566. 222),those who concerned the convenience to gain the medicine ( OR=119.652,95%CI:3.765-3802.184) and those who said that they could take medicine every day ( OR=88.245,95%CI:10.237-760.696) were more likely to accept PrEP. The subjects whose partners would be angry if they stick to use condoms( OR=0.106,95%CI:0.019-0.606),and those who suspected the effectiveness of drugs( OR=0.010,95%CI:0. 001-0.112) were less likely to accept PrEP. Conclusion The acceptability of PrEP is high in university students'MSM. The safety of drugs and the convenience to gain the medicine are the main influencing factors for usage of PrEP.Using of PrEP should be strengthened to the MSM.

Central Nervous System Fungal Infections ; complications ; Diabetic Ketoacidosis ; complications ; Humans ; Male ; Middle Aged ; Mucormycosis ; complications ; Nose Diseases ; complications

OBJECTIVE To explore the mechanism of Gαiand Gβγsubunits on dexmedetomidine (DMED)-induced sedation.METHODS Kunming mice were randomly placed into three groups(DMED group, DMED+dbcAMP/rolipram/gallein/M119 group, dbcAMP/rolipram/gallein/M119 group) to explore the regulation of dbcAMP/rolipram/gallein/M119 on DMED-induced sedation by establishing loss of righting reflex (LORR) model. DbcAMP/rolipram was intracerebroventricular injected and gallein/M119 was intraperitoneal injected 15 min before DMED intravenous injection. In CHO-α2A-AR cells, after administration of DMED/gallein/M119, the regulation on the cAMP accumulation stimulated by Forskolin (FSK) was detected, so was the intracellular calcium ion concentration ([Ca2+]i. The levels of pERK/pCREB were detected by Western Blot to explore the key signal molecules involved in DMED-induced sedation. RESULTS The ED50of DMED-induced LORR (200.0 nmol·kg-1) was increased to 375.0 or 433.3 nmol·kg-1by pre-treatment with cAMP analog dbcAMP(50 nmol/5μl per mouse)or phosphodies-terase 4 inhibitor rolipram(100 nmol/5μl per mouse).In addition,the ED50of DMED-induced LORR was decreased to 113.6 or 136.5 nmol·kg-1when pre-treated with Gβγsubunits inhibitor M119(100 mg·kg-1) or gallein(100 mg·kg-1)respectively.Administration of dbcAMP,rolipram,gallein or M119 alone had little effect on LORR of mice.Gallein(10 μmol·L-1)significantly inhibited forskolin-stimulated cAMP accumu-lation in CHO-α2A-AR cells.Compared with Gβγsubunits inhibitors or DMED alone,[Ca2+]iand pERK1/2 significantly increased after co-administration of Gβγsubunits inhibitors with DMED.DbcAMP(5 μmol·L-1) or rolipram (5 μmol·L- 1) alone had little effect on ERK1/2 phosphorylation, but decreased DMED-induced ERK1/2 phosphorylation after co-administration with DMED. Gβγsubunit inhibitors treatment increased DMED-induced phosphorylation of CREB, whereas dbcAMP or rolipram had little effect on pCREB induced by DMED.CONCLUSION Gβγsubunits might inhibit DMED-induced sedation through cAMP and pERK1/2 pathway,which was opposite to Gαisubuint.