Objective Recent researches show that bone sialoprotein is closely related with metastasis of breast cancer to bone. In order to investigate the effect of bone sialoprotein in the course of metastasis of breast cancer to bone, the following study was undertaken. Methods The growth curve, clone formation and adherence of breast cancer cells experiments were carried out to study the effect of hBSP over-expression on biological behaviors of breast cancer cells. Results It was found that breast cancer cells MDA-MB-231BO(pI) and MDA-MB-231BO(pID) stably transfected with hBSP cDNA were not inhibited through proliferation assay of breast cancer cells. The activity and tumorigenicity of breast cancer cells with hBSP over-expression were enhanced. Comprising with MDA-MB-231BO non-transfected the colony formation percentage of breast cancer cells MDA-MB-231BO(pI) and MDA-MB-231BO(pID) increased from 8.90%?0.53% to 15.6%?0.94% and 12.8%?0.77%. The OD_ 620 value of adherence of breast cancer cells MDA-MB-231BO(pI) and MDA-MB-231BO(pID)increased from 0.29?0.017 to 0.52?0.031 and 0.43?0.026 respectively in cells attachment assay, which suggested that over-expression of hBSP enhances breast cancer cell MDA-MB-231BO adherence and attachment property. Conclusion The study results suggested that hBSP over-expression had autocrine effect on breast cancer cells and could enhance the ability of breast cancer cells metastasizing to bone.

Diagnosis, Differential ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Male ; Prostatic Hyperplasia ; diagnosis ; Prostatic Intraepithelial Neoplasia ; diagnosis ; Prostatic Neoplasms ; diagnosis ; Racemases and Epimerases ; analysis ; Staining and Labeling ; methods

BACKGROUND: Bone sialoprotein (BSP) gene is expressed in human breast cancer cells, in which bone metastasis occurs easily outside the mineralized tissue. Clinical observation shows that the expression level of BSP of breast cancer cells at bone metastasis is higher that at the primary site;therefore, BSP may be closely related to tumor specific bone metastasis.The study on breast cancer bone metastasis can provide new drug target for clinical prevention and treatment.OBJECTIVE: To establish breast cancer cell strains of BSP with stable expression and observe the effect of BSP in the whole process of breast cancer bone metastasis.DESIGN: Controlled experiment.SETTING: College of Biological Sciences and Engineering, South China University of Science and Technology; Medical Experiment Center,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA.MATERIALS: This experiment was conducted in the Medical Experimental Center,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA,betweer November 2003 and March 2004..pIRES2-EGFP vector (5.3 kb) was purchased from BD Biosciences Clontech Inc.; E.Coli.Top10, pB-hBSP plasmid containing the coding region of hbsp, and human breast carcinoma cells, MDA-MB-231BR that was specifically transferred to brain and MDA -MB-231BO that was specifically transferred to bone.METHODS: hbsp gene was subcloned from pB-hBSP vector by PCR. Bg1Ⅱ and Pst Ⅰ restriction enzyme sites were inserted at 5' and 3' ends, orientation cloned to eukaryon expression vector pIRES2-EGFP, and constructed recombinant vector pIRES2-EGFP. The constructed recombinant vector was transfected into MDA-MB-231BR that was specifically transferred to brain and MDA-MB-231BO that was specifically transferred to bone.MAIN OUTCOME MEASURES: Construction of pIRES2-hBSP-EGFP recombinant expression vector; recombinant expression vector pIRES2-hBSP-EGFP transfecting breast cancer cells.Breast cancer strains specific in bone metastasis and brain metastasis were successfully transfected. The fluorescence labeling could be observed under the fluorescence microscope, and BSP had corresponding expression.CONCLUSION: The successful construction and transfection of pIRES2hBSP-EGFP of eukaryon expression vector would lay foundation for further study on the role of BSP in breast cancer metastasizing to bone in vivo or in vitro.

Objective: To compare the advantages and disadvantages of three staining methods of HBV. Methods: Normal Liver tissue and HBV-infected, HCV-infected ,or dually infected (HBV and HCV) liver tissues were selected for this study. Formalin- fixed, paraffin-embedded sections(4 ?m) were prepared. Each of the liver tissue specimens was detected by three staining methods, including immunohistochemical methods ,Shikata’s orcein stain and Victoria blue stain,respectively. Results: In the three methods , all of six HBV -infected cases showed intense staining, and three cases with dual infection (HBV and HCV) were weakly positive. However, both normal and HCV-infected liver tissues showed no staining. HBsAg stained dark brown with Immunohistochemical stain; HBsAg containing ground-glass hepatocytes stained magenta with Shikata’s orcein stain; HBsAg stained blue with Victoria blue. Conclusion: Each of three methods has its own advantages and disadvantages: high specificity and sensitivity, but high cost for immunohistochemical methods;complicated and overelabrate procedure for preparation of solutions, lower specificity and sensitivity,but low cost, for special staining methods.

ObjectiveTo observe the effect of buprenorphine sublingual tablets (Bup) and methadone for heroin addicts on out patients treatment during recovery period. MethodsHeroin addicts who completed acute drug detoxification treatment were assigned in the order of cross to the observation group and control group. The observation group and the control group were treated with Bup and methadone, respectively for six months. Both groups were observed on the time of keeping heroin abstinence during the treatment and were assessed with Anxiety Scale before and after Bup or methadone treatment. ResultsBup group kept abstinence from heroin longer than methadone group ( P ＜0.01 ) ,and had lower anxiety scores( P ＜0.05). ConclusionBupernorphine sublingual tablets used for out patients at recovery period could better alleviate heroin addicts' anxiety and prolong their abstinence time.

Objective: To study the effects of antisense epidermal growth factor receptor (EGFR) cDNA transfection on telomerase activity and telomere length of glioblastomas U87MG cells and the mechanism. Methods: Glioblastoma U87MG cells, which over-expressed EGFR, were transfected with antisense-EGFR cDNA constructs. Several clones stably expressing lower or undetectable levels of EGFR protein were obtained. The effect of antisense-EGFR cDNA on telomerase activity was assessed by Telomeric Repeat Amplification Protocol (TRAP) assay. The effect of antisense- EGFR cDNA on telomere length was determined by Southern blot hybridization. The mRNA expressions of hTERT, hTEP1 and c-myc were analyzed by semi-quantitative PCR. Results: U87MG cells that were transfected with antisense-EGFR cDNA expressed lower level of EGFR and were less responsive to the growth stimulation of EGF compared with the control cells. Telomerase activity was significantly decreased in the antisense-EGFR clones. Telomere length was shortened. The mRNA expression of hTERT was slightly decreased in the antisense-EGFR clones, whereas the expressions of hTEP1 and c-myc were not altered. Conclusion: Antisense-EGFR cDNA transfection can sufficiently inhibit EGFR signal transduction pathway, decrease telomerase activity and shorten telomere length, which may be a new mechanism of antisense-EGFR approach in tumor suppression. The down-regulation of hTERT mRNA may contribute to the decreased telomerase activity in the antisense-EGFR clones.

Hypertension is one of main risk factors for the occurrence and death of stroke and coronary heart disease. Its prevalence rate is rising year by year. It severely threatens the health of the human beings. The acupuncture method of "activating blood and dispersing wind, harmonizing Gan-Pi" for treating hypertension launched by Academician SHI Xue-min has aroused great attention due to good cur- ative effect and less adverse reactions. In this paper principles of the circular motion covered by the acupuncture method of "activating blood and dispersing wind, harmonizing Gan-Pi" were clarified.
Acupuncture ; Acupuncture Therapy ; methods ; Coronary Artery Disease ; Humans ; Hypertension ; therapy ; Medicine, Chinese Traditional ; Risk Factors ; Stroke ; Wind

We evaluated the patient in the early,advanced or healing phase of the disease by MRI in the treatment of chronic brucellosis spondylitis and to guide the clinical treatment.MRI images of 40 patients with clinically diagnosed chronic brucellosis spondylitis were analyzed retrospectively.The imaging findings of early,advanced and healing patients were summarized.MRI showed abnormal signals in the vertebral body,intervertebral disc,paraspinaI and psoas muscle.It is early stage if the intervertebral space was normal,and advanced stage if combined with interverbral space stenosis.It demonstrated short T1 and short T2 signal or similar to the normal vertebral body,combined with intervertebral space stenosis,for the healing stage.According to the specific imaging manifestations of chronic brucellar spondylitis in the course of disease development,it is possible to evaluate the clinical stage of the disease and guide the rational selection of clinical treatment.

Objective To investigate the age-associated changes of ultrastructure,mRNA and protein expressions of H+-K+-ATPase in elderly gastric parietal cell. Methods Fifty patients with relative normal stomach without gastroduodenal diseases were enrolled,including younger group (aged 20-59 years,n=19) and elderly group (aged≥60 years,n=31).Furthermore,the elderly group was divided into 3 subgroups:60-69 years old (n =11 ),70-79 years old (n=10 ),above 80 years old (n =10).The ultrastructure of gastric parietal cell was observed under electron microscope.The expression of H+-K+-ATPase α subunit mRNA and H+-K+-ATPase β subunit protein were assessed by quantitative real-time PCR and Western-blot,respectively.The ageing-associated changes of all these data were respectively compared. Results No significant difference was showed in the morphology of gastric parietal cell and acid-secretion-associated organelles among all the groups.The average ratio Am to Ac (Am means the area of mitochondria,Ac means the area of cytoplasm) of gastric parietal cell and the average At to Ac ratio (At means the area of secretory canaliculi and tubulovesicular system )between younger group and elderly group had no significant difference[(48.4±7.5) ％ vs.(50.6±7.6) ％,t=-0.775,P=0.444; (13.8±4.1) ％ vs.(12.2±4.7) ％,t=0.984,P=0.332].Meanwhile,there were no distinctions in the expression of H+-K+ -ATPase α subunit mRNA and H+-K+-ATPase protein among all elderly subgroups(F=1.522,2.32,P=0.24,0.114).However,the mRNA expression of H+-K+-ATPase a subunit was higher in the elderly group than in the younger group(t=-3.682,P=0.001).Furthermore,the expression of H+ -K+ -ATPase protein in the elderly group was increased as compared with younger group(t=-3.389,P=0.004). Conclusions Acidsecretion-associated organelles of human gastric parietal cell have no degeneration and the expression of H + -K+-ATPase is in trend of increase with aging,indicating that healthy elderly people have the basis of ultrastructure and molecular biology to maintain well function of acid secretion.

Objective To investigate the combination application of the intracranial buried electrode and electrical stimulation techniques in excising the epileptogenic zone in the central zone.Methods Seven patients with epileptogenic zone located close to or in the central zone of brain were recruited in the present study.The lone term ECoG monitoring and electrical stimulation of the codex were performed to identify the epileptogenic zone and the central zone of the brain after patients received intracranial electrode implants.The epileptogenic zone was excised with maximum preservation of the cen-tral zone.The patients were follow-up for 6 to 12 months,the outcomes were evaluated based on the Engel's scale and the Karnofsky(KPS)score.Results Seven patients did not experience any seizures and their Engei's and KPS scores were markedly improve after operation.Conclusions Intracranial buried electrodes and cortical electrical stimulation can guide the resection of epileptogenic zone in the central zone.Patients have no seizure and no serious dysfunction after operation and their quality of life was improved markedly.