Humans ; Neoplasms ; therapy ; Receptor, Epidermal Growth Factor

Objective To observe the expression of c-fos mRNA and protein in fluoride treated mouse fibroblast (FB) and osteoblast (OB) and to further explore the effects of c-fos in the osteogenic action of FB. Methods Mouse FB and OB were divided into control group and six fluoride groups (0, 0.0001, 0.0010, 0.1000, 1.0000, 10.0000,20.0000 mg/L F-), and the levels of c-fos protein at 2,4,24,48,72 h and c-fos mRNA at 48 h were measured by using ELISA and RT-PCR methods. Results Compared with the control group, fluoride increased the content of c-fos protein obviously in all FB group(P<0.01); and it is increased in 0.0001,0.0010 mg/L groups at 48 h (0.73±0.04, 0.64±0.14) and 0.0001 mg/L group at 72 h(0.70±0.17) in OB compared with the control group (0.32±0.04,0.27±0.05, P<0.01). Compared with the control group (0.95±0.11), RT-PCR revealed an increasing tendency of the expression of c-fos mRNA at 48 h in FB (1.06±0.16, 1.06±0.12,1.12±0.16,1.04±0.15,1.04±0.10,1.15±0.29), but the difference was not statistically significant(P>0.05); however, a statistically significant difference(P<0.01) of c-fos mRNA in 20.0000 mg/L group(1.40±0.17) in O B was found compared with the control group (1.06±0.06). Conclusion The higher expression of c-fos mRNA and protein in FB induced by fluoride may play an important role in the transformation of osteoblastic phenotype as well as increase the osteogenesis ability in FB.

Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Terminology as Topic ; Translations

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Eye ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Paralysis ; physiopathology ; therapy ; Trochlear Nerve Diseases ; physiopathology ; therapy ; Young Adult

Objective: To study the chemical constituents from Sinopodophyllum hexandrum and their antitumor activities. Methods: The constituents were separated by chromatography of silica gel, ODS, Sephadex LH20 and pre-TLC. Their structures were elucidated by spectroscopic means. The in vitro cytotoxic activities of the isolated compounds were studied by MTT method. Results: Nine compounds were isolated and identified as 8,2'-diprenylquercetin 3-methyl ether-4'-O-β-D-glucoside (1), 8,2'-diprenyl quercetin-3- methylether (2), 5,7,4'-trihydroxy-3'-(3-methylbut-2-enyl)-3-methoxy flavone (3), 8-prenylkaempferol (4), sophoflavescenol (5), podoverine A (6), sinoflavonoid K (7), diosmetin (8) and acacetin (9). Conclusion: Compound 1 is a new compound named sinoflavonoid glycosides A, and compounds 5-9 are isolated from S. hexandrum for the first time. Compounds 1-5 show cytotoxicities against HeLa cells with IC50 of 42.6, 46.9, 26.9, 16.1 and 31.2 μmol/L, respectively.

Objective To study the relationship of urine RBC morphology between UF-100 urine sediment analytic instrument andphase contrast microscope.Methods The UF-100 urine sediment analytic instrument to analyze 500 urine specimens and study the relation-ship of urine RBC morphology between urine sediment analytic instrument and phase contrast microscope.Results The according perceptionof Normocytic,Microcytic and Non-classified RBC between phase contrast microscope and UF-100 urine sediment analytic instrument RBC-info are 91.4%,94.4%,83.3% respectively,the according perception between phase contrast microscope and RBC-P70Fsc are 94.9%,95.7%,94.7% respectively,and the according perception between phase contrast microscope and RBC Fsc-DW are 84.4%,86.8%,90.5% respectively,the specificity of UF-100 and phase contrast microscope in glomerular hematuria and non-glomerular hematuria are84.3%,88.1% and 83.3%,87.9% respectively.Conclusion The results show that the UF-100 urine sediment analytic instrument issimply operating,fast and high accurate,and which can instruct clinical dignose,therapy and prognosis judgement.

Objective To establish the reference interval for serum prostate-specific antigen (PSA) in apparent healthy men of different ages in Nanjing.Methods A total of 25 820 healthy men undergoing routine physical examinations in the First Affiliated Hospital of Nanjing Medical University from October 2013 to September 2015 were selected for the study.All of them were screened by prostate B ultrasound,excluding abnormal urinary tract diseases.The concentration of serum PSA and free prostate-specific antigen (fPSA) were measured by automatic luminescence immunoassay analyzer,and the fPSA/PSA values were calculated.The participants were divided into four groups (20～ 39,40～ 59,60～ 79 and older than 80 years old groups),then the median,5th,25th,75th and 95th percentiles of both PSA and fPSA/PSA were counted,respectively.Results The median of PSA (95th percentile ranges) of these groups by age from low to high were 0.78 (1.93),0.90 (2.93),1.34(6.60) and 2.01(11.91),respectively.The 25th to 75th percentiles were 0.55～1.11,0.61～1.36,0.77～2.51 and 0.94 ～ 4.19,respectively.The median of fPSA/PSA (95 th percentile ranges) were 0.37 (0.60),0.31 (0.56),0.28 (0.53) and 0.29(0.52),respectively.The 25th to 75th percentiles were 0.28～0.46,0.23～0.40,0.22～0.36 and 0.23～ 0.37,respectively.Among all the groups,median differences of both PSA and fPSA/PSA were statistically significant (P＜0.05),and PSA levels rise with age.PSA levels in different regions were different.Conclusion The PSA level of men under 40 years in Nanjing should be 0～2.5 ng/ml,40～60 years should be 0～4 ng/ml,while men who are above 60 years,could use 0～5 ng/ml as reference interval.

Objective To explore the effect of Bailing capsule on epithelial-mesenchymal transition( EMT) in rats with adenine-in-duced tubulointerstitial fibrosis. Methods Tubulointerstitial fibrosis animal models were established and SD rats were divided into mo-del group ( n = 30), treatment group ( n = 30) and control group( n = 30), randomly. Experimental rats were harvested at 7 w, 12 w,17 w after onset of experiment and functional evaluations were performed. Histology, immunohistology were examined to investigateboth histolopathology changes and the expression of bone morphogenic protein-7 (BMP-7), transforming growth factor-β1 (TGF-β1 )and a-smooth muscle actin (α-SMA) in kidneys at three time points mentioned above, respectively. Results Compared with controlgroup, 24 h urinary protein in model group lost increasingly and significantly difference appeared at three time points relative to controlgroup ( P ＜ 0.01 ). Urinary NAG in model group was markedly higher than that in control group from 7 w after onset (P ＜ 0.01 ) andwas increasingly raised at 12 w and 17 w (P＜0.01). The value of blood BUN and Cr in model group increased at 7 w (P＞0.05) rel-ative to control group. There was significant difference at 12 w and 17.w (P ＜ 0.01 ). Histologically, kidneys in model group, at 7 w,exhibited tubular casts and gently tubular dilation, granuloma in cortex, mononuclear cells infiltration in tubulointerstitial areas, andmild interstitial fibrosis. At 12 w, the degree of tubular injury and tubulointerstitial fibrosis gradually aggravated. Up to 17 w, diffusetubular dilation or atrophy was observed and focal tubules disappear. Diffuse interstitial fibrosis was exhibited. In normal kidneys, im-munohistochemistry suggested that the light expression of BMP-7 was detected in proximal renal tubular epithelial cells and marked ex-pression was identified in distal tubule, collecting duct, and renal tubular epithelial in junction area between cortex and medulla. How-ever, the expression of BMP-7 in kidneys of model group significantly decreased with increasing tubulointerstitial fibrosis and was nega-tive correlation with the expression of TGF-β1(r = -0. 981 P＜0.01) and α-SMA (r= -0.975 P＜0.01). Bailing capsule ad-ministration protected the expression of BMP-7 and reduced TGF-β1 and α-SMA expression before 12 w(P＜ 0.01 ). Conclusions Ourstudy shows an anti-fibrotic reno-protective function of Bailing capsule in rats with tubulointerstitial fibrosis via prevention of epithelial-mesenchymal transition at early stage. However, the beneficial effect lost with increasing tubulointerstitial fibrosis.

Objective To investigate the different diagnostic value of serum Chitinase 3-like 1 protein(CHI3L1)and Alpha-fe-toprotein(AFP)in diagnosing hepatocellular carcinoma (HCC).Methods One hundreds HCC patients confirmed by histopa-thology were recruited between December,2015 to April,2016 from the First Affiliated Hospital of Nanjing Medical Univer-sity.Simultaneously,100 patients with chronic hepatitis B and 59 healthy individuals,matched by sex and age with HCC pa-tients,were recruited as control groups.Serum CHI3L1and AFP were measured in different groups and the difference were analyzed by STATA 1 2.0 Statistical software.The ability of these two items in differentiating different group was analyzing by ROC curve using MedCal Ver 15.2.2 software.Results Serum CHI3L1 were significantly differences in the three groups using Kruskal-Wallis analysis (χ2=93.19,P=0.000),the differences were further compared in different two groups using Mann-Whitney analysis.The results showed that serum CHI3L1 in HCC group were significantly higher than in chronic hepatitis B and healthy control group (P=0.000,z=8.766,7.400).Serum AFP were significantly differences in the three groups using Kruskal-Wallis analysis (χ2=147.54,P=0.000),and the differences were further compared in different two groups using Mann-Whitney analysis.The results showed that serum AFP in HCC group were significantly higher than in chronic hepatitis B and healthy control group (P=0.000,z=10.938,9.033).The ROC curve analysis of serum CHI3L1 and AFP for differentiating HCC group from CHB group showed that CHI3L1 yield AUC of 0.859 (95% CI:0.803~0.904) with 85% sensitivity,79% specificity and 76.8 pg/ml cut-off value,AFP yield AUC of 0.948 (95% CI:0.904~0.974)with 85% sensitivity,98% specificity and 7.6 ng/ml cut-off value,in distinguishing HCC with CHB group,the power of AFP was superior to that of CHI3L1 (P=0.006).The ROC curve analysis of serum CHI3L1 and AFP for differentiating HCC group from healthy individuals group showed that CHI3L1 yield AUC of 0.852 (95% CI:0.787~0.903)with 85% sensi-tivity,76% specificity and 76.8 pg/ml cut-off value,AFP yield AUC of 0.929 (95% CI:0.878~0.964)with 84% sensitivi-ty,100% specificity and 7.8 ng/ml cut-off value,in distinguishing HCC with healthy individuals group,the power of AFP was also superior to that of CHI3L1 (P=0.045).Conclusion Serum CHI3L1 similar to AFP has much power ability to di-agnosis HCC,but AFP was superior to CHI3L1.

Objective:To remove murine embryonic stem cells(mESC)from the differentiating cell culture and purify the differentiated cells by Magnetic Activated Cell Sorting(MACS).Methods:Neural differentiation of mESC was induced by a 5-stage method.The specific cell surface marker,SSEA-1,was used to identify ES cells in the differentiating cells.The optimal dilutions of mouse anti mouse SSEA-1 IgM primary antibody and FITC conjugated goat anti mouse secondary antibody were determined before the flow cytometry test.The incubation time and incubation temperature of primary antibody were all optimized to make the cytometry test accurate.After the optimization,stage 4 cells were dissociated into single cell suspension,incubated with antibody of SSEA-1 and microbeads conjugated goat anti mouse IgM,and then sorted through the magnetic field.The rate of SSEA-1 positive cells in pre-and post-separation groups was assessed by flow cytometry,and the viability of cells was evaluated by trypan blue staining counting under light microscopy.Results:The proportion of SSEA-1 positive cells in the separated cells can be reduced from(7.19?1.36)% to(1.34?0.80)%.The survival rate of sorted cells was more than 92%,similar to that of pre-separation cells.Conclusions:The MACS system we used can effectively remove mESC from the differentiated cells.The sorted cells will be well provided for the subsequent studies about transplantation therapy.