Objective To determine the estrogenic activity of dibutyl phthalate DBP. Methods The tested compound was dibutyl phthalate. Human estrogen-dependent MCF-7 breast cancer cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum FBS. Five days before the addition of the tested compounds the cells were rinsed by phosphate balanced solution PBS and the medium was substituted with a phenol red-free RPMI 1640 medium containing 5% dextral charcoal-stripped FBS. The respective tested compound was added in fresh medium and the control cells received only the vehicle ethanol. The proliferation of MCF-7 cells was analyzed by the MTT assay growth curves mitotic index and coloning efficiency. Results Compared with the control the proliferation of tested cells treated with DBP like estradiol was markedly enhanced and the activity of the cell proliferation reached the maximum at 10-5 mol/L DBP. During log phase the mitotic index of the test cells treated with DBP and estradiol was significantly increased. The cell coloning efficiency was enhanced which was treated by 10-5 mol/L DBP only for 48 hours. The results showed the time-dependent and dose-dependent model. Conclusion Dibutyl phthalate may enhance the proliferation of human breast cancer MCF-7 cells in vitro that demonstrates an estrogenic activity of dibutyl phthalate.

OBJECTIVETo explore clinical significance of rotational axis of distal femur on Chinese adults in total knee arthroplasty (TKA).

METHODSThere were 86 Chinese adults (106 normal knees) including 47 males (53 knees) and 39 females (53 knees), 54 knees were on left and 52 on right. The CT scan was employed in the distal femur. The scan direction was aligned to be on the plane perpendicular to the mechanical axis of the femoral. The CT images of cross sections across lateral and medial femoral epicondyle were moved to personal computer,lateral angle between anterior posterior line (APL) and surgical transepicondylar axis (STEA) (ATA),lateral angle between posterior condylar line (PCL) and APL (APA), angle between perpendicularity of APL and PCL (A-PA), posterior condylar angle (PCA), condylar twist angle (CTA), angle between clinical transepicondylar axis (CTEA) and STEA (CSA) were measured. These values were divided into different groups according to gender and side, the values of CTA, PCA, A-PA, angle PT (varus of tibia plateau), constant 3, ATA, APA and constant 90° were compared by statistically. A-PA and PCA, and CTA were analysed statistically with the liner regression, the relationship among CTEA, STEA ,PCL, APL and PLP were performed to assess by liner regression.

RESULTSATA was (89.79 ± 1.22)°, APA was (84.84 ± 1.83)°, A-PA was (5.16 ± 1.83)°, PCA was (4.80 ± 1.23)°, CTA was (8.23 ± 1.40), CSA was (3.45 ± 0.68)°. All the parameters had no differences on sex and side,but CSA had difference on male and female. There was no difference among angle PT, PCA, A-PA. There was significant difference in CAT, constant 30 and angle PT, PCA,A-PA. There was no difference between ATA and constant 90°, but there was difference between APA and constant 90°. There was relativity between PCA and CTA, and also PCA and A-PA, CTA and A-PA. There was significant relativity between STEA and CTEA, between STEA and APL, between STEA and PCL, and also between APLP, APL and PCL, but there was no significant relativity between PCL and CTEA.

CONCLUSIONTKA for Chinese, the section of femoral posterior condyle should be external 5° to obtain the optimum rotational orientation. The property is different entirely between STEA and CTEA, the rotational alignment is not performed according to parallel to the CTEA in distal femur. Among STEA, APL, PCL, the STEA is the most reliable mark ofrotational alignment of the distal femur, and the PCL is the less reliable mark.

Adult ; Arthroplasty, Replacement, Knee ; Female ; Femur ; anatomy & histology ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Rotation ; Tomography, X-Ray Computed

Aim To study relative bioavailablity of cefaclor effervescent tablets in healthy volunteers. Methods According to the crossover design, A volunteers were each orally given a single does of the 0.75 g cefaclor effervescent tablets and cefaclor capsules with an interval of 5 days between the two formulations.The plasma concentrations of the drug were determined by RP-HPLC.Pharmacokinetic parameters were obtained by ATPK programe,and calculated on the basis of open single compartment model.Results After a single oral dose, the peak levels in plasma averaged Cmax(31.27?5.81)?g?ml-1 and(30.56?5.25) ?g?ml-1 at (0.58?0.12)h and(0.73?0.17)h and AUC0～4(35.48?4.65) ?g?h?ml-1 and (35.89?2.90) ?g?h?ml-1 for tablet and capsule,respectively. Conclusion The result shows that two formulations are bioequivalence.

Objective To observe the effects of 75 gram glucose oral tolerance test (75 g OGTT) and standard mixed meal test (SMMT) on insulin secretion function of the islets of Langerhans and plasma free fatty acid (FFA) in patients with type 2 diabetes mellitus.Methods Seventy-six patients with type 2 diabetes without using insulin and with no obvious complications were recruited for 75 g OGTT following overnight fasting on the first day and SMMT (bread 50 g,egg 50 g and milk 250 ml) on the 7th day.Blood specimens were collected from each patients before the tests and 30 min,60 min,120 min and 180 min after glucose or meal load to measure their levels of plasma glucose,serum insulin,C peptide,FFA and lipids (total cholesterol,triglyceride,high-density and low-density lipoprotein cholesterol).Results No difference in fasting plasma glucose,serum insulin,C peptide,FFA and lipids between 75 g OGTT and SMMT was found.Postprandial plasma glucose 30 min,60 min,120 min and 180 min after 75 g OGTT was significantly higher than that after SMMT,with (15.3?3.5) vs (9.9?3.4) mmol/L,(18.2?4.8) vs (12.8?4.0) mmol/L,(16.3?5.8) vs (12.2?4.9) mmol/L and (10.6?5.4) vs (9.5?4.5) mmol/L (F=28.1,P

OBJECTIVE: To select specific DNA aptamer for determining ketamine by FluMag-SELEX. METHODS: Based on magnetic beads with tosyl surface modification as solid carrier and ketamine as target, a random ssDNA library with total length of 78 bp in vitro was compounded. After 13 rounds screening, DNA cloning and sequencing were done. Primary and secondary, structures were analyzed. The affinity, specificity and Kd values of selected aptamer were measured by monitoring the fluorescence intensity. RESULTS: Two ssDNA aptamers (Apt#4 and Apt#8) were successfully selected with high and specific abilities to bind ketamine as target with Kd value of 0.59 and 0.66 μmol/L. The prediction of secondary structure was main stem-loop and G-tetramer. The stem was the basis of stability of aptamer's structure. And loop and G-tetramer was the key of specific binding of ketamine. CONCLUSION FluMag-SELEX can greatly improve the selection efficiency of the aptamer, obtain the ketamine-binding DNA aptamer, and develop a new method for rapid detection of ketamine.
Aptamers, Nucleotide/metabolism* ; DNA ; DNA, Single-Stranded/genetics* ; In Vitro Techniques ; Ketamine/metabolism* ; Oligonucleotides ; SELEX Aptamer Technique/methods*

Objective：The aim of this study was to determine the expression of p21waf1, c erbB 2, and p53 in node negative breast cancer in relation to the clinicopathological parameters and prognosis. Methods： Expression of p21waf1, c erbB 2 and p53 in 121 patients were determined by LSAB immunohistochemical method, and Kaplan Meier method and Cox proportional hazard model were used to analyze the relationship of their expressions with prognosis. Results： (1) The expression rate of p21waf1 was 48.8％ , and was in connection with histological grade and ER positive. (2) The expression rate of p53 and c erbB 2 was 36.4％ and 26.4％ respectively. C erbB 2 positive expression was associated with histological grade. (3) the expression of p21waf1 was inversely correlated with that of p53 protein (P< 0.05). (4) the results of survival analysis with Kaplan Meier method showed that better disease free survival(DFS) in the patients with p21waf1 expression than those without, the negative c erbB 2 expression exhibited a significantly better prognosis than positive c erbB 2 expression (P< 0.01). Conclusions： There was a significant correlation between the expression of p21waf1, c erbB 2,and tumor grade; a p53 indenpent pathway can mainly induce the expression of p21waf1; p21waf1 expression was a good prognostic marker for the patients with node negative breast cancer; c erbB 2 positive expression was an independent prognostic marker.

OBJECTIVETo observe the occurrence and progression of liver fibrosis induced by pig serum exposure and bile duct ligation, and analyze the relationship between hepatic inflammation and liver fibrosis.

METHODSChronically immune-mediated liver fibrosis was induced in rats by weekly injection of pig serum (IPS) into the peritoneal cavity at 3 ml/kg for 12 weeks. Cholestatic fibrosis was induced by common bile duct ligation (BDL). The Knodell score was used to evaluate the histological changes in the liver, and immunohistochemistry was performed using anti-SMA, anti-ED1, anti-CK7, and anti-CD45 antibodies. Quantitative real time PCR (qPCR) analysis was employed to quantify the mRNA expression of the genes related to inflammation, including interleukin-1beta (IL-1beta), IL-6, monocyte chemotactic protein-1, tumor necrosis factor-alpha, regulated upon activation normal T cell expressed and secreted (RANTES), transforming growth factor-beta, platelet-derived growth factor A, as well as the genes associated with fibrogenesis, namely collagen 1, alphaSMA, MMP-9 and TIMP-1.

RESULTSKnodell scores for periportal necrosis, intralobular degeneration and focal necrosis, and portal inflammation were all significantly higher in the BDL group than in the IPS group (P<0.01), whereas the scores for fibrosis was higher in the IPS group (P<0.05). Immunohistochemistry showed obvious inflammation with numerous alphaSMA-positive cells in the liver of the rats in BDL group; the liver of the rats in IPS group showed numerous alphaSMA-positive myofibroblasts with limited inflammatory cell infiltration. qPCR demonstrated a significant up-regulation of the genes related to extracellular matrix remodeling such as collagen 1 (P<0.01), alphaSMA (P<0.01), MMP-9 (P<0.01) and TIMP-1 (P<0.01) in the rat liver in IPS group compared with those in the normal control group, and the mRNA expressions of the inflammation-related cytokines, except for RANTES, were comparable with those in the control. In contrast, the BDL group showed a significant up-regulation of all the pro-inflammatory genes examined with also increased expression of the fibrogenesis-related genes (P<0.05).

CONCLUSIONLiver fibrosis induced by IPS is characterized by active ECM remodeling in the absence of obvious inflammation, indicating that chronic development of liver fibrosis can be independent of active hepatic inflammation. BDL-induced liver fibrosis highlights obvious inflammation and fibrous proliferation in the liver.

Animals ; Bile Ducts ; surgery ; Cholestasis ; complications ; physiopathology ; Ligation ; Liver Cirrhosis, Experimental ; etiology ; immunology ; pathology ; Male ; Rats ; Rats, Inbred F344 ; Serum ; immunology ; Swine

Objective To investigate the relationship between cortical watershed infarction and carotid artery stenosis and evaluate the stent insertion operation.Methods After 23 cortical watershed infarction patients diagnosed by CT or MRI received DSA detection,we performed stent insertion operationon 11 patients according to their requirements,and conservative treatment on the remaining 12 patients.All the patients underwent follow up for 6-12 months post-operatively.Results Among the 23 cortical watershed infarction patients,22 Were detected with carotid artery stenosis.Statistical analysis showed that the degree of carotid artery stenosis was associated With the elinical svmDtoms and the volume of steal phenomenon(P<0.05);further,the artery stenosis improvement was over 90%with the stent inserted;conversely,dizziness and steal phenomenon disappeared.The post procedure follow-up,ranging 6-12 months,showed that the patients with stent insertion got less new symptoms,steal phenomenon and artery stenosis,compared with the patients with conservation treatment(P<0.05).Conclusion Cortical watershed infarction is associated with carotid artery stenosis.The stent insertion iS useful for the treatment ofcarotid artery stenosis and prevention of cortical watershed infarction.

Polyamidoamine-polyethylene glycol (PAMAM-PEG) copolymers were synthesized using IPDI as coupling reagent by two-step method. The copolymers were characterized by IR spectrum and 1H NMR spectrum, and the PEG conjugating ratios of the copolymers were calculated equal to 10% and 30% separately. MTT assay indicated that after PEGylation a lower cytotoxicity of the copolymers could be found, and with increasing PEG conjugating ratio the cytotoxicity decreased obviously. Agarose gel retardation assay demonstrated that PAMAM-PEG copolymers could be combined with DNA and PAMAM-PEG/DNA complexes were prepared by self-assembly. DLS measurement showed that when N/P > or = 50, the particle size of copolymer/ gene complexes was in a range of 150-200 nm, and the zeta potential was in a range of 10-25 mV. In vitro gene transfection illustrated that when N/P < or = 50, the gene transfection efficiency of PAMAM-PEG copolymers was a little less than that of PAMAM-G5, but the transfection efficiency can be raised by increasing N/P ratio or transfection time. Considering both cytotoxicity and transfection efficiency aspects PAMAM-PEG-13 was more effect than PAMAM-PEG-39 in PEGylation.
Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Survival ; drug effects ; DNA ; chemistry ; pharmacology ; Dendrimers ; chemical synthesis ; pharmacology ; Gene Transfer Techniques ; Genetic Vectors ; Humans ; Isocyanates ; chemistry ; Liver Neoplasms ; pathology ; Particle Size ; Polyamines ; chemistry ; Polyethylene Glycols ; chemical synthesis ; chemistry ; pharmacology ; Transfection

OBJECTIVETo investigate the effects of metoprolol on electrophysiology of ischemic and anoxic myocardium in diabetic rats.

METHODSForty Sprague-Dawley (SD) rats were divided into 4 groups: diabetes group; diabetes and ablation of left sympathetic nerve group; diabetes and metoprolol group and sham group. The diabetes model was induced by intraperitoneal injection of streptozotocin (STZ, 60 mg/kg). The ventricular diastolic effective threshold (DET), effective refractive period (ERP), and Ventricular fibrillation threshold (VFT) were measured. The serum concentration of nerve growth factor (NGF) was measured.

RESULTSMetoprolol increased DET of ischemic and anoxic myocardium in diabetic rats. The ablation of the left sympathetic nerve increased VFT of diabetic rats. VFT in metoprolo group was significantly increased compared to diabetes group after ischemia. The concentrations of NGF in diabetic group and metoprolol group were higher than those in sham group. There were no difference in NGF levels between ablation of left sympathetic nerve group and sham group.

CONCLUSIONThe remodeling of sympathetic nerve affects the electrophysiology of ischemic myocardium of diabetic rats. Metoprolol can increase the VFT and decrease the excitation threshold of the ischemic myocardium in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; physiopathology ; Heart ; drug effects ; physiopathology ; Male ; Metoprolol ; pharmacology ; Myocardial Ischemia ; physiopathology ; Nerve Growth Factor ; blood ; Rats ; Rats, Sprague-Dawley ; Sympathectomy