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.

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.

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.

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

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.

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 assess the levels of nucleosomes released from peripheral blood mononu-clear cells(PBMC)of patients with systemic lupus erythematosus(SLE),and their relationship with auto-antibodies as well as disease activity.Methods Levels of both nucleosomes released from PBMC and vari-ous auto-antibodies were detected by ELISA in sera from SLE patients.The disease severity was evaluated using SLEDAI(systemic lupus erythematosus disease activity index)system.Results Levels of nucleosomes released from PBMC were significantly higher in patients with active SLE than those of patients with inactive disease and normal controls(39.39?25.70,13.44?8.82,and11.73?7.87IU/mL,respectively).There was a significant positive correlation between nucleosome levels and SLEDAI scores,serum ds-DNA auto-an-tibody levels,and low C3levels.Conclusion Nucleosomes released from apoptotic PBMC of patients with SLE is closely correlated to disease activity,which implies that nucleosomes may play an important role in the pathogenesis of SLE.

Objective To investigate the role of nucleosome in the pathogenesis of systemic lupus erythematosus (SLE). Methods BALB/c mice were immunized with nucleosome, and then serum dsDNA and ANA autoantibodies were detected by ELISA. Kidney specimens were observed by immunofluorescence and histological examination. Results High titers of IgG dsDNA and ANA autoantibodies in sera of BALB/c mice were observed at the 14th day after immunization with nucleosome. Nephritis and immune complex deposition in renal glomeruli were observed at the 35th day. Conclusion Nucleosome could induce SLE-like disease in BALB/c mice, and may play a critical role in the pathogenesis of SLE.

Objective To investigate specific cellular immune response induced by dendritic cells(DCs) activated by human papilloma virus (HPV) 16 E7 peptide. Methods DCs separated from peripheral blood mononuclear cells were induced with granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4), the DCs were then activated by HPV 16 E7 peptide. After that the surface antigens on the DCs were detected by flow cytometer. Homogenic mixed lymphocyte reaction (HMLR) with DCs was performed. LDH release assay was also used to detect the activity of cytotoxic T lymphocytes(CTL)to Caski tumor cells, which was induced by DCs. Results The surface antigens on the activated DCs were highly expressed, such as CD1a(58.4 ? 6.7)%, CD80(70.6 ? 3.4)% and HLA-DR(74.8 ? 4.2)%. HMLR showed that the activated DCs stimulated T cell proliferation significantly. Antigen-specific CTL induced by the activated DCs specifically killed Caski tumer cells, but had no effect on SiHa and AN3CA cells. Conclusions DCs loaded with HPV 16 E7 peptide can induce highly effective and specific cellular immune response.