Objective To investigate the effect of arsenic trioxide on expression VEGF of lymphoma cells. Methods The VEGF mRNA expression was analysed by by Real-time PCR, and VEGF protein expression in Raji and Jurkat lymphoma cell lines by ELISA. Results ATO can inhibit lymphoma cells by inducing apoptosis. ATO induced lymphoma cell apoptosis was due to time.With the period of ATO effecting on cells goes, the expression of VEGF mRNA and the protein were down-regulated significantly (after 24, 48, 72 h). There were, the VEGF mRNA △△Ct data of treated with ATO, at 12 h, for Raji: 0.75±0.15, 72 h, Jurkat: 1.67±0.13. After 72 h, Raji: 8.95±0.38; Jurkat: 9.09±0.16 (f =3.54, P <0.01; t =3.65, P <0.01). And about the VEGF protein, at 12 h, Raji: 198.38±4.37; Jurkat: 563.11±3.81. After 72 h, Raji: 23.55±2.06; Jurkat: 57.11 ±3.88 (t =2.48, P <0.05; t =2.59, P <0.05). Conclusion ATO can inhibit the proliferation of lymphoma cells by down-regulating the expression of VEGF mRNA and its protein.

Objective To establish a simple and practical method for isolating and purifying nucleosomes. Methods Nuclei were isolated from chicken erythrocytes, and then digested with staphylococcal nuclease. After centrifugation, the supernatant of digestion was separated and centrifugated on sucrose gradients. Results Nucleosomes with good stability were isolated properly by gradient centrifugation. Conclusion This method for the isolation and purification of nucleosomes is simple and effective, which might contribute to the further researches of the roles of nucleosomes in the pathogenesis of systemic lupus erythematosus.

Background Blepharitis is a common ocular surface disease.It is associated with the disorder of lipid secretion of meibomian gland.The change of tear film stability can cause dry eye symptoms,so blepharitis is thought to be one of the factors causing dry eye,but the relation between them is in study. Objective This study was to observe the morphology of meibomian gland in blepharitis patients and to investigate the correlation of morphology of meibomian gland with dry eye. Methods A series of case-observational study was designed in this study.A total of 83 eyes of consecutive 83 blepharitis patients were enrolled in Henan Eye Institute from October 2010 to April 2011.Blepharitis was diagnosed based on American Preferred Practice Pattern Guidelines.Some relevant ocular examinations were performed under the informed consent of the subjects,including the anterior segment manifestation by the slit lamp,such as meibography,lid margin abnormality,and the dry eye-relevant examinations,such as tear film break-up time (BUT),Schirmer test Ⅰ and corneal fluorescein staining also been carried out.Tear film shape was examined by film interference images and scored.Absent degree of meibomian gland was graded under a Noncontact Infrared Meibography.The correlations of absent degree of meibomian gland with ocular syndrome score,dry eye examination results were evaluated using Spearman rank correlation coefficients.Informed consent was obtained prior to this trail. Results No significant difference in the frequencies of blepharitis was found between male and female among different ages (x2 =2.69,P =0.75 ).Absent grading of the meibomian glands was positively correlated with age of blepharitis patients ( r =0.58,P =0.00 ),lid margin abnormality scores ( r =0.64,P =0.00 ),conjuntival hyperemia score ( r=0.50,P =0.00),tear film interference imaging grade ( r =0.23,P =0.04 ),corneal fluorescein staining score( r =0.50,P =0.00 ) but was negatively correlated with BUT ( r =-0.32,P =0.00 ).No significant correlation was found between meibography grading and gender( r =-0.09 ; P =0.99 ) or Schirmer test Ⅰ ( r =-0.05;P =0.69 ).No significant difference was found in meibography grading between male and female in different age groups(Z=-0.09,P=0.93). Conclusions Blepharitis can irriter dry eye symptom because of overevaporation of tear fluid and abnormality of secreting function of meibomian glands.The missing of the meibomain glands increases with age in the patients with blepharitis.Noncontact Meibography System is an assistant tool to the diagnosis of blepharitis.

Objective:To study the effects of polysaccharide from Lycium barbarum (LBP) on cell apoptosis and bc1-2 gene expression in irradiated mice. Methods:Forty eight Kunming mice were divided into three groups (LBP group, radiation group and normal group). LBP chow was prepared through adding 0.8%LBP to normal chow and was supplied to LBP group. Normal chow was supplied to normal group and radiation group. LBP group and radiation group were exposed to whole-body 60Co ?-rays at the dose of 0.084 Gy/day for 6 w, five times a week and the total dose was 2.52Gy. Then the micronucleus frequency of polychromatic erythrocytes(MF),chromosome and sperm aberration frequency, caspase-3 mRNA expression, cell apoptosis and bcl-2 gene expression were detected. Results:LBP could significantly lower MF, chromosome and sperm aberration frequency, and cell apoptosis , and it could increase the proliferation activity of bone marrow cells and bc1-2 gene expression in irradiated mice and decrease caspase-3 mRNA expression.Conclusion:The radioprotective effect of LBP is related to regulation of cell apoptosis and bc1-2 gene expression.

A series of novel 4-substituted-3-nitrobenzamide derivatives were designed and synthesized. The structures of the target compounds were confirmed with 1H NMR, 13C NMR, MS and element analysis. Anti-tumor activities against HCT-116, MDA-MB435 and HL-60 cell lines in vitro were evaluated by SRB assay. The results indicated most of the target compounds exhibited potent anti-tumor activity. Compound 4a showed the most potent inhibitory activities against three cancer cell lines with the GI50 values of 1.904-2.111 micromol x L(-1). Compounds 4g, 41-4n exhibited more potent inhibitory activities against MDA-MB435 and HL-60 cell lines with the GI50 values of 1.008-3.586 micromol x L(-1) and 1.993-3.778 micromol x L(-1), respectively. The structure-activity relationship of these compounds is discussed preliminarily.
Antineoplastic Agents ; chemical synthesis ; pharmacology ; Benzamides ; chemical synthesis ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Drug Design ; HL-60 Cells ; Humans ; Inhibitory Concentration 50 ; Structure-Activity Relationship

Background Human limbal allograft transplantation or limbal autograft transplantation are the primary approaches to the severe corneal-blindness,but their application in clinic were limited because of the defects of donor material.With the development of tissue engineering technology,transplantation of in vitro cultured limbal epithelial stem cells is being an advanced management.Objective The aim of this work was to expand human limbal epithelial stem cells ex vivo under the guidance of confocal microscope and to lay the foundation for fabricating ex vivo cultured cell sheets.Methods Ten eyes of ten patients were examined with the Heidelberg Retina Tomography Ⅲ Rostock Cornea Module(HRT3-RCM)to elucidate the structure of the human corneoscleral limbus and to correlate limbal epithelial dimensions.According to the analysis of the images of limbal epithelia,the limbal tissues provided by Eye Bank of Henan Eye Institute were cut into suitable explants.Then,this study was conducted to expand limbal epithelial stem cells ex vivo on denuded amniotic membrane.The phenotypes of primary cultured cells were evaluated by morphology and immunofluorescent staining with antibodies for limbal epithelial stem cell markers (p63,cytokeratinl9)and differentiation markers(keratin 3,involucrin).This experimental procedure was approved by the Ethic Committee of Henan Provincial People's Hospital.The written informed consent was obtained from subjects before initiation of any examination.Results The palisade morphology of human limbus was imaged clearly on the laser scanning in vivo confocal microscopy and many hyperreflective cells were observed in palisade basal cells.The cell-island phenomenon was seen in the basement membrane under the laser scanning in vivo confocal microscopy.The oblique sections of limbus showed many papilla-like epithelial columns below the superficial limbal epithelia.Throughout the experiment duration,the epithelial cells grew well with the migration rates from limbal tissue (68.62± 16.94)％ and the migration time(5.83 ±2.04)days,which depended on the tissue freshness.Compared with the second and forth batch of tissue,the migration rates of the third and sixth batch of tissues were significantly higher(P＜0.05),and the migration time was evidently longer in the forth and sixth batch of tissue compared with the first,second,third and fifth batch(P＜0.05).The positively expressing rates in the cultured corneal stem cells were 4.05％ and 36.52％ for p63,26.07％ and 40.55％ for CK19,57.88％ and 40.81％ for K3,64.66％ and 59.19％ for involucrin.Conclusion Human limbal epithelial stem cells can be successfully and purposefully obtained from the limbal tissue based on the guidance confocal miscroscope.The cultured corneal stem cells can grow well on the denuded amniotic membrane

Objective To investigate the safety of rituximab combination chemotherapy in the treatment of B-cell non-Hodgkin' s lymphoma (B-NHL) complicated with hepatitis B virus (HBV) infection,and assess the incidence of HBV reactivation reduced by prophylactic lamivudine.Methods A retrospective study of HBV-related markers,HBV-DNA and liver function was performed before and after rituximabcontaining treatment in B-NHL patients.Thirty nine B-NHL patients with HBcAb(+)/HBsAb(-) were divided into prophylactic group (14 cases) and control group (25 cases).The incidences of HBV reactivation,functional damage of liver were measured.Results Among the 108 B-NHL patients who received rituximab combinatio nchemotherapy,15 (13.89 ％) were HBsAg (+) and 39 (36.11％) HBsAg (-) / HBcAb (+).Of the 15 HBsAg (+)patients,2 (13.3 ％) experienced reactivation of HBV.The prevalence of HBV reactivation was 7.7 ％(1/13) in patients who received prophylactic antiviral treatment and 50 ％ (1/2) in those who did not receivelamivudine.Among the 39 HBsAg (-) / HBcAb (+) patients,3 cases (7.7 ％) experienced reactivation of HBV.The prevalence of HBV reactivation was 0 in patients who receivcd prophylactic lamivudine treatment and 12 ％ (3/25) in those who did not receive this antiviral drug.Conclusion Prophylactic lamivudine before rituximab combination chemotherapy can reduce HBV reactivation obviously.

Objective To prepare controlled release microspheres of vascular endothelial growth factor(VEGF)& calcium alginate and observe their effect on proliferation of human umbilical vein endo- thelial cells(HUVEC)in order to provide theoretical basis for controlled release of VEGF facilitating an- giogenesis of tissue engineering bone.Methods VEGF-calcium alginate microspheres were prepared by using the needle extrusion/external gelation method to investigate physicochemical character and in vitro release of VEGF.According to the different ingredients added into the culture medium,the seconda- ry cultured HUVEC were divided into four groups,ie,control group,microsphere group,VEGF group and VEGF-calcium alginate microsphere group,in which the proliferation of the cultured HUVEC was ob- served with cell counting method,MTT method and flow cytometry.Results The calcium alginate mi- crospheres were revealed as spherical shape and evenly distributed,with mean grain diameter of(560?50)?m,carrying capacity of 0.72 ng/mg and the encapsulation efficiency of 54%.Smooth controlled re- lease in VEGF-alginate microspheres lasted for more than 10 days.Proliferation of the cultured HUVEC was accelerated the most in VEGF group at the beginning but in EGF-calcium alginate microsphere group at midanaphase compared with other groups,with statistical difference(P＜0.05).There was no statis- tical difference upon cell counting,cell activity and time point of cell cycle between control group and mi- crosphere group.Conclusion VEGF-sodium alginate microapheres can continue activity of VEGF,re- lease VEGF for over 10 days and promote proliferation cultured HUVEC for a long time.

?AIM: To explore the relation of homocysteine ( Hcy ) and its metabolic related indicators with primary angle-closure glaucoma ( PACG) .?METHODS: In this study, a total of 150 PACG patients and 150 controls were enrolled. The patients with PACG were diagnosed by applanation tonometer, Humphrey perimetry ( HVF ) , optical coherence tomography ( OCT ) , gonioscope. The normal controls were recruited from physical examination center. Blood samples were collected and the plasma was used to determine homocysteine, vitamin B12 and folic acid. DNA was extracted to determine the methylenetetrahydrofolate reductase ( MTHFR ) C677TT genotype and gene polymorphism.?RESULTS:There was a statistically significant difference (t=2. 04,P=0. 04) in mean homocysteine levels between patients with PACG ( 16. 11 ± 1. 66μmol/L ) and controls (15. 74± 1. 52μmol/L). The level of vitamin B12 in PACG group was 248. 09 ± 119. 07pg/mL and the control group was 230. 21 ± 142. 93pg/mL. No statistically significant difference was found on the vitamin B12 level between the two groups ( t = 0. 84, P = 0. 40 ). The folic acid levels showed no statistically significant differences( t=1. 65,P=0.17) between the PACG group(12. 02±7. 81ng/mL) and the control group (13. 15 ± 6. 25ng/mL). The frequency distribution of the MTHFR C677T genotypes was found to be significantly associated (χ2 =6. 2,P<0. 05) with PCAG (CC 84. 7%, CT 4. 7%, TT 10. 7%) as compared to the controls (CC 94. 7%, CT 2. 0%, TT 3. 3%). The frequency distribution of TT genotypes of MTHFR C677T in the PACG group was significantly higher than that in the control group. Moreover, our findings showed significant difference (χ2=14. 2,P<0. 05) between the allele frequency of the C677T single nucleotide polymorphism within the MTHFR gene in POAG patients ( T 87. 0%, C 13. 0%) and the control group(T 95. 7%,C 4. 3%) .?CONCLUSION: These results show that the Hcy level and the TT genotypes of MTHFR C677T were higher in the PACG patients than in controls. Therefore, our data suggests that high Hcy level and the related indicators are associated with high PACG risk.

Objective To develop a human immune-deficiency virus (HIV) infection risk appraisal model suitable for men who has sex with men (MSM) in Guangzhou,and to provide tools for follow-up the outcomes on health education and behavior intervention.Methods A cros-sectional study was conducted in Guangzhou from 2008 to 2010.Based on the HIV surveillance data,the main risk factors of HIV infection among MSM were screened by means of logistic regression.Degree on relative risk was transformed into risk scores by adopting the statistics models.Individual risk scores,group risk scores and individual infection risk in comparison with usual MSM groups could then be calculated according to the rate of exposure on those risk factors appeared in data from the surveillance programs.Results Risk factors related to HIV infection among MSM and the quantitative assessment standard (risk scores and risk scores table of population groups) for those factors were set up by multiple logistic regression,including age,location of registered residence,monthly income,major location for finding their sexual partners,HIV testing in the past year,age when having the first sexual intercourse,rate of condom use in the past six months,symptoms related to sexually transmitted diseases (STDs) and syphilis in particular.The average risk score of population was 6.06,with risk scores for HIV positive and negative as 3.10 and 18.08 respectively (P＜ 0.001 ).The rates of H IV infection for different score groups were 0.9％,2.0％,7.0％,14.4％ and 33.3％,respectively.The sensitivity and specificity on the prediction of scores were 54.4％ and 75.4％ respectively,with the accuracy rate as 74.2％.Conclusion HIV infection risk model could be used to quantify and classify the individual' s infectious status and related factors among MSM more directly and effectively,so as to help the individuals to identify their high-risk behaviors as well as lifestyles.We felt that it could also serve as an important tool used for personalized HIV health education and behavior intervention programs.