Objective To study the influence of fluorine on signaling pathway of osteoprotegerin(OPG)/ receptor activator of NF-κB ligand(RANKL) in cultured rat osteoblasts.Methods Osteoblasts were isolated from skull of neonatal rats(＜ 24 hours) by enzyme digestion,and fluorine of different concentrations [0 (control),1 × 10-3,1 × 10-4,1 × 10-5,1 × l0-6 and 1 × 10-7 mol/L] were added into the culture medium of second generation of osteoblasts.The expressions of OPG and RANKL mRNA were determined using real-time PCR 24 and 48 hours after culturing.The expressions of OPG and RANKL protein were measured by Western blotting.Results ① After exposed to fluorine for 24 hours,the differences of RANKL and OPG mRNA expression had statistical significance between groups(F =30.95,22.62,all P ＜ 0.01),the expression of RANKL mRNA(5.99 ± 0.39) in the 1 × 10-5 mol/L group and the expressions of OPG mRNA(3.52 ± 0.09,4.81 ± 0.15,3.68 ± 0.04) in the 1 × 10-4,1 × 10-5 and 1 × 10-6 mol/L groups were higher than those of the control group(3.20 ± 0.19,3.09 ± 0.58,all P ＜ 0.05),but in the 1 × 10-3 mol/L group,RANKL mRNA(2.29 ± 0.18) was lower than that of the control group(P ＜ 0.05).After exposed to fluorine for 48 hours,the differences of RANKL and OPG mRNA expression had statistical significance between groups(F =26.62,5.72,all P ＜ 0.01),the expressions of RANKL and OPG mRNA(6.67 ± 0.49 and 5.05 ± 0.51) in the 1 × 10-5 mol/L group were higher than those of the control group(4.29 ± 0.07 and 4.34 ± 0.12,all P ＜ 0.05),and in the 1 × 10-3 mol/L group the expression of OPG mRNA(3.63 ± 0.49) was lower than that of the control group(P ＜ 0.05).② The expression of RANKL protein was not statistically significant between 24 hours and 48 hours groups (F =0.07,0.49,all P ＞ 0.05) ; the differences of OPG protein expression had statistical significance between groups(F =3.26,P ＜ 0.05),the expression of OPG protein in the 1 × 10-5 mol/L group(1.45 ± 0.10) was higher than that of the control group(1.05 ± 0.06,P ＜ 0.05) at the 24 hours.After 48 hours,the expression of OPG protein was not statistically significant(F =0.44,P ＞ 0.05).Conclusions At lower fluorine concentrations,bone formation is the main activity.But when fluorine concentration increased and time prolonged,the osteoclast differentiation and maturation are promoted,and the bone resorption is the main thing.

Objective To investigate the epidemic characteristics and trends of incidence and mortality of prostate cancer in cancer registration areas in Zhejiang province from 2000 to 2009.Methods Clinical data incidence and mortality of prostate cancer were collected from 6 cancer registration areas in Zhejiang province,including Hangzhou,Jiaxing,Jiashan,Haining,Shangyu and Xianju.Crude rates,standardized rate and change trend,age-specific rates and annual percent change (APC,95％ CI) of prostate cancer were checked,sorted and analyzed in Zhejiang Cancer Center.Results The prostate cancer incidence rate from 2000 to 2009 was 9.79/100 000,age-standardized incidence rates by world standard population (ASIRW) was 6.39/100 000,and the incidence cumulative risk of males aged 0-74 was 0.72％ ; while the mortality rate was 2.73/100 000,age-standardized mortality rates by world standard population was 1.74/100 000,and the mortality cumulative risk of males aged 0-74 was 0.14％.Age-specific incidence of prostate cancer remained low before 50,years old and peaked at over 85-year-old group (130.30/100 000).Age-specific mortality of prostate cancer increased after 55,and also peaked at over 85-year-old group (81.19/100 000).The annual prostate cancer incidence rate generally grew from 1.39/100 000 (2000) to 13.89/100 000 (2009),and the APC was 14.18％ (95％ CI,9.68％-18.98％).Meanwhile,the prostate cancer mortality rate also increased from 1.52/100 000 (2000) to 3.58/100 000 (2009),and the APC was 11.83％ (95％ CI,5.69％-18.33％).Conclusion Prostate cancer incidence and mortality in Zhejiang cancer registration areas increased sharply,and the prevention and treatment of prostate cancer should be strengthened.

Objective: To obtain purified and functional CDNF-his recombinant protein and prepare its polyclonal antibodies.Methods:Preparation of recombinant CDNF-his was carried out in HEK 293 T cells with pVR1012-CDNF-his successfully constructed transfected into them.Then,the recombinant protein was purified by Ni-NTA immunoaffinity chromatography.The purity was analyzed by SDS-PAGE and the protein′s identity was tested by Western blot.MTT was used to verify the biological function of the protein purified.New Zealand white rabbits were immunized with purified CDNF-his protein for preparation of polyclonal antibodies.Results:pVR1012-CDNF-his expressed successfully in HEK 293 T cells.The purity of protein was up to more than 90%after purification.MTT showed that CDNF-his was able to protect PC 12 cells from damage by 6-OHDA.The polyclonal antibody was detected at the end of animal immunizing process.Conclusion: A method to express and purify protein using HEK 293T cell and following Ni-NTA immunoaffinity chromatography has been built.CDNF-his with biological activity is obtained based that.Finally, polyclonal antibodies of CDNF were generated successfully.

Objective To explore the changes and the tendency of breast cancer radiotherapy in China within the past decade.Methods A survey on breast cancer radiotherapy in 1 999 and 2006 was conducted in 4 hospitals located within the southeast coastal region of China,including patients'clinical characteristics,the purpose of radiotherapy and its techniques.Results The percentage of breast conservative treatment(BCT)increased from 3%in 1999 to 13%in 2006.For radiotherapy techniques in 2006 compared with 1999,the growth trend was found in use of breast immobilization devices(80% vs.46%),treatment planning system(70% vs.23%),and CT simulation(14%vs.0).The frequency was increased in the chest wall irradiation after masteetomy(90%vs.67%),but decreased in internal mammary region(30%vs.76%)and axilla(37%vs.69%)treatment.There were no differences in total dose and fractionation prescription.Conclusions BCT was performed more frequently,but postmastectomy radiotherapy was still essential option in China at present.Although some advanced techniques were applied frequently in clinical practice.Simulation,treatment planning,and irradiation target design were urgent for improvement and standardization.

Objective To analyze the associations of the interleukin-10 (IL-10) promoter-1082G/A and-819C/T single nucleotide polymorphism (SNP) and serum level of IL-10 with intracranial aneurysm (IAs). Methods The polymerase chain reaction (PCR) and DNA direct sequencing methods were used to detect IL-10 gene promoter district two SNP site,-1082G/A and-819C/T genotype frequency and allele frequency in 206 patients with IAs and 187 controls. Chi-square test was used to analyze differences between two groups. The serum level of IL-10 was analyzed by ELISA, and t-test was used to analyze significant differences between two groups. Results There were significant differences in genotypes of GG and GA+AA, as well as the alleles G and A, in-1082G/A locus between IAs group and control group (P<0.01). There were higher frequencies of genotype GA+AA and the allele A in IAs group than those in control group (P<0.01). There was higher risk of suffering IAs in patients with genotype GA+AA (OR=4.137, 95%CI:2.476-6.914) and the allele A (OR=3.368, 95%CI:2.476-4.583). There were higher frequencies of the genotype CT+TT and the allele T in-819C/T locus in IAs group than those of control group (P<0.01). There was higher risk of suffering IAs in patients with genotype CT+TT (OR=3.393, 95%CI:1.952-5.900) and the allele T (OR=3.764, 95%CI:2.730-5.192). The serum level of IL-10 was significantly lower in IAs group than that of control group (P<0.01). Conclusion The IL-10 promoter SNP influences the expression of IL-10. IL-10 promoter-1082G/A and-819C/T polymorphisms are correlated with the formation of IAs.

The proliferation difference between Human Umbilical Vein Endothelial Cells (HUVEC) and Human Umbilical Artery Smooth Muscle Cells (HUASMC) in response to the concentration of silver nanoparticles was investigated via MTT, BCA and FCM tests. The obtained experimental data were statistically analyzed and discussed in order to know the causation of the proliferation difference. The results show there is significant difference in proliferation between HUVEC and HUASMC corresponding to the concentration of silver nanoparticles, and such difference can be attributed to the varied adhesion shape and apoptosis of the cells being influenced by nano-Ag content and Fetal bovine serum (FBS) content in culture medium.
Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Metal Nanoparticles ; chemistry ; Muscle, Smooth, Vascular ; cytology ; Silver ; pharmacology ; Umbilical Arteries ; cytology

Objective To investigate the differences of measurement of gross target volume (GTV)between endoscopic ultrasonography ( EUS )-based ( GTVEUS ) and computed tomography ( CT ) -based (GTVCT) method for thoracic esophageal squamous cell carcinoma. Methods EUS was performed on 36consecutive patients with thoracic squamous cell carcinoma, and the superior and inferior boarders of the tumor defined by EUS were marked with hemoclips. The CT planning scan was then performed with the patient in supine position, and the GTVCT and GTVEUS were contoured respectively. The lengths ( LCT and LEUS) and spatial locations of longitudinal GTVCT and GTVEUS were compared. Results The mean LCT and LEUS were (7. 79 ± 3. 15 ) cm and (7. 42 ± 2. 72) cm, respectively ( t = 0. 82, P ＞ 0. 05 ), with a correlation coefficient of 0. 61 (P ＜0. 001 ). Locations of longitudinal GTVCT and GTVEUS were compared in 34cases, with 2 excluded for invisualization on CT. The mean conformal index (CI) was (0. 79 ± 0. 18 ), and spatial variations were found in 71% patients, with 8 patients at proximal end and 21 others at distal end.There was no clip placement associated complication. Conclusion Endoscopic hemoclips placement is safe and reliable. EUS can provide additional information to CT in defining longitudinal GTV in thoracic esophageal squamous cell carcinoma, especially in superficial and submucosal carcinomas.

Objective:To investigate the potential association between FADS1 rs1 74537 polymorphism and serum proteins in patients with aggressive periodontitis,which may provide benefits for diagnosis and treatment of aggressive periodontitis.Methods:A total of 353 patients with aggressive periodontitis (group AgP)and 1 25 matched controls (group HP)were recruited in the study.Genotyping of FADS1 rs1 74537 and serum biochemical indexes were tested at the study’s start.The relationships between the levels of TP,GLB,ALB,A/G and genotyping were analyzed.Results:(1 )The detection rate of allele G in group AgP was higher than that in group HP(68.1% vs.61 .2%,P=0.046,OR=1 .35,95% CI 1 .00-1 .83 );the detection rate of genotype GG in group AgP was higher than in group HP(45 .5%vs. 34.4%,P=0.029,OR=1 .60,95%CI 1 .05 -2.44).(2)In group AgP,the patients with GG geno-type exhibited significantly lower TP,GLB than the patients with GT+TT genotype [(77.08 ±7.88)g/L vs.(79.00 ±4.66)g/L,P=0.007;(28.1 7 ±7.63)g/L vs.(29.88 ±3.49)g/L,P=0.007)and the higher A/G(1 .72 ±0.22 vs.1 .67 ±0.22,P=0.040),but there was no significant difference in ALB between the patients with GG genotype and the patients with GT+TT genotype.In group HP,there were no significant differences in TP,GLB,A/G and ALB between individuals with genotype GT+TT and with genotype GG.(3 )Compared with individuals with genotype GT+TT in group HP,the AgP pa-tients with genotype GT +TT exhibited significantly higher TP,GLB [(79.00 ±4.66 ) g/L vs. (75.20 ±4.53)g/L,P<0.01;(29.88 ±3.49)g/L vs.(26.55 ±2.94)g/L,P<0.01 )and the lo-wer A/G(1 .67 ±0.22 vs.1 .88 ±0.30,P<0.01 ),but there was no significant difference in ALB. There were no significant differences in TP,GLB,A/G and ALB the between the AgP patients with ge-notype GG and the healthy subjects with the same genotype either.Conclusion:FADS1 rs1 74537 poly-morphism is associated with aggressive periodontitis.The patients with genotype GG in group AgP had relatively lower TP,GLB and higher A/G.Genotype GG might be a risk indicator for aggressive periodon-titis by reducing host defense capability and contributing to inflammatory response in the occurrence and development of aggressive periodontitis.

Objective To explore the effect of the combined sequential application of er-lotinib between the chemotherapy periods on the treatment of the patients with advanced non small cell lung cancer (NSCLC).Methods 45 patients with advanced NSCLC were collected.The pa-tients with the initial treatment were treated with gemcitabine combined with platinum for chemotherapy,while the patients with retreatment were treated with docetaxel or pemetrexed for chemotherapy.Between the chemotherapy periods,the sequential erlotinib therapy was applied to all the patients.The short-term efficacy and the adverse reactions were evaluated and compared. Results The clinical efficacy of the initial treatment patients and the retreatment patients were 27.6% and 18.8% respectively (P >0.05),and the clinical control rate of the initial treatment patients was significantly higher than that of the retreatment patients (P <0.05).The clinical ef-ficacy of the patients with a history of smoking was significantly lower than that of the patients without a history of smoking (P <0.05).The clinical efficacy of the initial treatment patients with age over 65 years old was significantly higher than that of the initial treatment patients with age less than 65 years old(P <0.05).The clinical efficacy of the initial treatment patients with stage Ⅲ b was significantly higher than that of the initial treatment patients with stage Ⅳ (P <0.05).The clinical efficacy of the retreatment patients with adenocarcinoma pathological type was significantly higher than that of the patients without adenocarcinoma pathological type (P <0 .0 5 ).The main adverse reactions were nausea ,diarrhea ,rash ,vomiting and loss of appetite .
Conclusion The clinical efficacy of the combined sequential application of erlotinib between the chemotherapy periods in the treatment of the patients with advanced NSCLC is not such high. Smoking history,age,tumor stage and pathological type may affect its curative effect.Efficacy of this therapy in controlling the disease progress of the initial treatment patients is significant,the adverse reactions are mild,and the tolerance of the patients for the therapy is higher.

Objective To observe and analyze the curative effects of the combined sequential application of erlotinib between the chemotherapy periods in the treatment of the patients with ad-vanced non small cell lung cancer (NSCLC).Methods 45 patients with advanced NSCLC were se-lected as the research objects.The patients of the initial treatment were treated with gemcitabine combined with platinum for chemotherapy and the patients of retreatment were treated with doc-etaxel or pemetrexed for chemotherapy.Between the chemotherapy periods,the sequential erlotinib therapy was applied to all the patients.The short-term curative effects and the adverse reactions were evaluated and compared.Results The clinical efficiency of the initial treatment patients and the retreatment patients were 27.6% and 18.8% respectively.There were no significant differences of short-term curative effects and the clinical efficiency between the initial treatment patients and the retreatment patients (P >0.05),while the clinical control rate of the initial treatment patients was significantly higher than that of the retreatment patients (P <0.05).The clinical efficiency of the patients with a smoking history was significantly lower than that of the patients without smok-ing history(P <0.05).The clinical efficiency of the initial treatment patients with age of 65 years or older was significantly higher than that of the initial treatment patients of age < 6 5 years (P <0.05).The clinical efficiency of the initial treatment patients with stage Ⅲb was signifi-cantly higher than that of the initial treatment patients with stage Ⅳ(P <0.05).The clinical ef-ficiency of the retreatment patients with adenocarcinoma pathological type was significantly higher than that of the patients without adenocarcinoma pathological type (P <0.05).The main adverse reactions of the patients were nausea,diarrhea,rash,vomiting,loss of appetite.Conclusion The clinical efficiency of the combined sequential application of erlotinib between the chemothera-py periods in the treatment of the patients with advanced NSCLC is not high.Smoking history, age,tumor stage,pathological type may affect its curative effect.The effects of this therapy in controlling the disease progress of the initial treatment patients are more significant,the adverse reactions are mild,and the tolerance of the patients for the therapy is higher.