Animals ; Chemical and Drug Induced Liver Injury ; metabolism ; pathology ; prevention & control ; Disease Models, Animal ; Female ; Genistein ; therapeutic use ; Liver ; drug effects ; metabolism ; pathology ; Male ; Mice

OBJECTIVETo explore the effects of cyclopamine on the proliferation and apoptosis of LNCaP cells and the expression of the PCA3 gene in human prostate cancer in vitro.

METHODSLNCaP cells were treated with cyclopamine at the concentrations of 1, 5, 10 and 15 micromol/L for 24, 48 and 72 hours. The inhibitory effects of cyclopamine on the proliferation and apoptosis of the LNCaP cells were detected by MTT and flow cytometry respectively, the morphological changes of the cells observed by Hoechst 33258 staining, and the expression of the PCA3 gene determined by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction (FQ-RT-PCR).

RESULTSCompared with the blank control group, cyclopamine significantly inhibited the proliferation of the LNCaP cells at 5, 10 and 15 micromol/L (P <0.01), reaching IC50 at 10 micro mol/L at 48 hours. The apoptosis rates of the LNCaP cells at 24, 48 and 72 hours were 37.21%, 57.38% and 57.98% in the 10 micromol/L group and 21. 16% , 71.31% and 72.90% in the 15 micro.mol/L group, significantly different from those in the control (P <0. 01). The cell apoptosis showed a rising trend with the increase of cyclopamine concentration and acting-time, while the expression of the PCA3 gene was decreasing with the increased concentration of cyclopamine, significantly lower than that of the blank control group (P <0.01) , and extremely low in the 10 micromo/L group

CONCLUSIONCyclopamine intervention at 10 and 15 micromol/L for 48 and 72 hours could significantly inhibit the at all time points. Proliferation and induce the apoptosis of LNCaP cells and reduce the expression level of PCA3.

Antigens, Neoplasm ; genetics ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Male ; Prostatic Neoplasms ; genetics ; pathology ; Veratrum Alkaloids ; pharmacology

Objective To explore the clinical and pathologic features of Langerhans cell histiocytosis(LCH).Method Thirty-eight cases of LCH from both in hospital and outpatient department in dermatology and pediatry department from Jan.1991 to Jun.2006 were analyzed about the features of clinic,pathology and immunohistochemistry.Results The mean age of onset was 2.18 years old.The male /female ratio was 1.71.Skin lesions occurred in 78.9% of the patients.Among them,68.5% were as the first manifestation.The eruptions mainly distributed on trunk,90% of them presented as hemorrhagic maculopapules,nevertheless,3.3% of the eruptions showed as crust with hilar depression,which was similar to pityriasis lichenoides etvarioliformis acuta.Fever,hepatomegalia and splenomegia occurred in patients at a rate of 60.5%,68.4%,55.3% respectively.Thirty-one point six percent of the patients had got lymphadenectasis,the neck and inguinal lymph nodes were the common site to be affected.Ossature involvement occurred in 31.6% of the patients,which 8.3% got multiple injuries,howe-ver,91.7% got a solitary bone involved.Skull was the main site to be injured,else were lumbar,humerus,hipbone,an so on.Respiratory tract,auditory canal,mucosa were also the sites involved in this disease,but the incidence rate was lower than 10%,respectively.The laboratory data showed that 81.3% of the patients were anaemia,60.7% with abnormal subgroup of T-cell,and 32.1% positive for EBV-IgG.The skin histopathology data of 26/30 cases revealed that lichenoid infiltrates of Langerhans cells confined to the upper dermis.Cytologic features were cells with abundant eosinophilic cytoplasms,and longitudinally grooved or reniform nuclei.Lymphocytes and a few eosinophils also could be seen.Four cases of thirty showed that the proliferative Langerhans cells were with pale cytoplasms,besides,there were numerous eosinophils,and sometimes a few multinucleate cells were scattered.The immunity test of 20 cases of thirty displayed that CD1a(+)S100(+)KP-1(-).Biopsy of lymphaden and tumor of the skull of the rest 8 patients were all diagnosed as eosinophilic granuloma through both hematoxylin and eosin-stained section and immune marks.Conclusions Multiple systems can be involved in LCH.Hemorrhagic maculopapules,fever and splenohepatomegalia are common presentations in this disease.The morphous of nucles of histiocytes is particular,and to diagnose definitely,both CD1a(+) and S-100(+) are needed.

Objective To observe the expression of vascular endothelial growth factor(VEGF) mRNA and protein in fluoride(F~-) treated fibroblast(FB) of mice in planar(2D) and FBs populated collagen lattice(3D) culture systems and to further explore the effects of VEGF on the osteogenic action of FB. Methods FB were divided into 0 (control group), 0.0001,0.0010,0.1000,1.0000,10.0000 and 20.0000 mg/L groups(F~-). The levels of VEGF mRNA and protein at 48 h were measured by using RT-PCR, ELISA and immunohistochemistry (IHC) methods. Results The expression of VEGF mRNA increased obviously in group of 0.1000 mg/L(1.08 ± 0.09) in 3D FB compared with the control group(0.93 ± 0.02, all P < 0.05). Fluoride increased the content of VEGF protein obviously in groups of 0.1000,1.0000,10.0000 mg/L(0.19 ± 0.02, 0.26 ± 0.01 and 0.32 ± 0.01 ), higher than that in 2D FB culture supematant in the control group(0.14 ± 0.01, all P < 0.05) ; and in groups of 0.1000, 1.0000 rag/L(0.59 ± 0.06 and 0.52 ± 0.03) it was higher than that in 3D FB culture supematant in the control group(0.37 ± 0.05, all P< 0.01 ). The IHC results showed that the VEGF positive staining cells increased significantly in group of 0.001 mg/L (0.45 ± 0.05) in 2D FB when it was compared with control group(0.36 ± 0.03, P< 0.05); and in groups of 0.0010, 0.1000, 1.0000 rag/L(0.62 ± 0.04,0.70 ± 0.06 and 0.65 ± 0.07) are it was higher than that in 3D FB control group (0.44 ± 0.04, P < 0.05 or < 0.01 ). Conclusions The higher expression of VEGF mRNA and protein in 2D and 3D FB induced by fluoride may play an important role in stimulating the osteogenesis ability in FB.

Animals ; Antigens, CD ; metabolism ; Apoptosis ; B7-2 Antigen ; metabolism ; CD4-CD8 Ratio ; Dendritic Cells ; metabolism ; pathology ; Disease Models, Animal ; Lectins, C-Type ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Minor Histocompatibility Antigens ; Multiple Organ Failure ; metabolism ; pathology ; Receptors, Cell Surface ; metabolism ; Spleen ; metabolism ; pathology

Objective To explore the role and clinical significance of interleukin-6(IL-6) in children with bronchial asthma.Methods Sera were collected from 29 cases with asthmatic attacks,32 asthmatic children who in stable conditions,and 20 health children.Serum IL-6 concentrations were measured by radioimmunoassay(RIA).Results 1.Asthmatic children appeared significantly higher levels of serum IL-6 during asthmatic attacks as compared to those in stable conditions and healthy ones respectively(P3 years old had significantly higher serum IL6 concentrations than ≤3 years old children in remission.Conclusion IL-6 may participate pathogenesis of asthma,and it may play different biological roles during asthmatic attacksas or stable conditions.

Objective To investigate the anti-tumor effect of ZM-66 on multidrug-resistant leukemic cell line K562/ADM.
Methods The K562/ADM cells were treated with varying concentrations (0, 1, 2, 4×10-3 mmol/L) of ZM-66 or etoposide for 24 hours. The proliferation was detected by Sulforhodamine B Sodium Salt (SRB) assay and apoptosis was detected by flow cytometry analysis and fluorescent staining. In addition, the expression levels of p53 and bax genes in K562/ADM cells were detected by RT-PCR analysis. The level of P-glycoprotein (P-gp), P53 and Bax protein in K562/ADM cells were detected by Western blot assay.
Results SRB assay demonstrated that etoposide had little inhibitory effect on K562/ADM cells, whereas ZM-66 (1, 2, 4×10-3 mmol/L) had significantly inhibitory effect on K562/ADM cells (all P<0.01). The acridine orange/propidium iodide dual staining showed that there were typical condensation of chromatin and nuclear fragmentation nuclei with red color in ZM-66 treated cells. Flow cytometric analysis showed that there was a significantly increase of apoptotic cells in K562/ADM cells after treated with ZM-66. RT-PCR showed that the p53 and bax mRNA expression levels in K562/ADM cells treated with ZM-66 at 1, 2, 4×10-3 mmol/L were higher than those in the cell without treatment. Western blot showed that the P53 and Bax protein expression levels in K562/ADM cells treated with ZM-66 at 2, 4×10-3 mmol/L were higher than those in the cell without treatment. But the P-gp protein expression level in K562/ADM cells treated with ZM-66 at 2, 4×10-3 mmol/L was gradually lower than those in the cell without treatment.
Conclusion ZM-66 is able to induce cell death by apoptosis in vitro, as a result of the reverse of the apoptosis resistance in drug-resistant K562/ADM cells by modulating expression of key factors associated with apoptosis induction.

Objective To investigate the impact of anesthetization and manipulation on neurotransmitters glutamate and dopamine during cell transplantation. Methods Neural stem cells cultured in vitro from postnatal rats were implanted into the striatum of normal adult rats. Brain microdialysis combined with high-performance liquid chromatography (HPLC) was applied to dynamically detect the impact of microdialysis probe implantation, anesthetics ketamine and pentobarbital and implanted cells etc on glutamate and dopamine levels. Results After 15 min probe implantation into the rat striatum, glutamate and dopamine levels in the striatum increased, evidently higher than the baseline value and declined to the baseline level within about 5-6 h. Ketamine and pentobarbital anesthetization for 15 rain resulted in a transient increase in glutamate and dopamine levels in the rat striamm;there were no significant effects on in vivo glutamate and dopamine levels by cell implantation itself. Conclusion Routine doses of intraperitoneal ketamine or pentobarbital anesthetization may result in a transient increase in glutamate and dopamine levels within the brain extracellular space. Based on these data, the optimal time for commencing brain microdialysis on glutamate and dopamine should be at least 6 h after probe implantation.

Introduction:The 2010 targets of the China Hepatitis B Prevention Programme were a prevalence of hepatitis B surface antigen (HBsAg) less than 1.0% for children less than five years old and less than 6.0% for the total population. This survey assessed the prevalence of Hepatitis B infection in Lianyungang, Jiangsu province, China in 2009–2010.Methods:Multistage sampling was used with 2372 subjects among 17 selected villages. Blood specimen collection and testing by enzyme-linked immunosorbnet assay (ELISA) were completed using the following markers for hepatitis infection: HBsAg and antibody to HBsAg (anti-HBs); hepatitis B e antigen (HBeAg) and antibody to HBeAg (anti-HBe); and hepatitis B core antibody (total anti-HBc). The data were analysed with Epi Info, version 3.3.2.Results:The prevalence of HBsAg was 2.4% (95% Confidence Interval [CI]: 1.8–3.0; Adjusted Prevalence [AP] 2.9%); anti-HBs prevalence was 51.1% (95% CI: 49.1–53.1; AP 49.2%) and total anti-HBc prevalence was 41.7% (95% CI: 39.8–43.7; AP 45.5%). The prevalence of HBsAg and total anti-HBc positivity increased from young to older age groups, yet the prevalence of anti-HBs positivity decreased from young to older age groups (P< 0.001 for all). There was no difference in the prevalences of HBsAg and anti-HBs among females and males (P= 0.108 and 0.089), but females had a higher prevalence than males for total anti-HBc positivity (P< 0.001). Discussion: This survey showed that in 2010 the prevalence of HBsAg among children aged less than five years was lower than the national target of 1.0% and that the prevalence of HBsAg for the total population was lower than the national target of 6.0%.