Objective To develop a highly sensitive and accurate time-resolved immunofluorometric assay (TR-IFMA) for measurement of Dickkopf-1(DKK-1),as a novel serologic biomarker for lung cancer. Methods The study constructed a two-manoclonal-antibody "sandwich"-type assay and the sensitivity, within run CV and between run CV and accuracy were evaluated. Serum DKK-1 concentrations were measured by TR-IFMA in 120 healthy controls, in 72 benign lung disease patients, and in 212 lung cancer patients before surgery. The association between serum DKK-1 levels and clinicopathological features were evaluated. Results A standard curve for DKK-1 TR-IFMA had been developed with good sensitivity (0.08 μg/L). Both within run CV and between run CV were less than 6.5%. Accuracy studies, parallelism and precision data were determined and all found to be satisfactory. The validity of DKK-1 assay was confirmed by the good correlation between the results obtained by TR-IFMA and commercial ELISA (r=0.972, P=0.01). The serum levels of DKK-1 were higher in lung cancer patients 31.93(79.47-18.03) μg/L than in benign lung diseases 15.25(18.41-11.49) μg/L and in healthy controls 13. 90( 16. 91-11.02) μg/L DKK-1 levels were significant associated with the presence of distant metastases, as well as lymph node metastases and TNM stage, but not with patients' age, gender and tumor histology. At the cutoff of 22.63 μg/L, the diagnostic sensitivity, specificity, accuracy, positive predictive value and negative predictive value of the TR-IFMA for lung carcinoma were 68.4%, 92.2%, 82.1% ,90.6% and 72.5%. Diagnostic sensitivity and accuracy were higher for small cell carcinoma than for non-small cell carcinoma (70.7% vs 69.5% and 85.6% vs 80.7%, respectively). Conclusions A highly sensitive and reliable TR-IFMA for DKK-1 has been developed. The determination of serum DKK-1 levels may be useful for diagnosis and tumor staging of lung cancer.

CYP2D6 is one of the cytochrome P450 isozymes which are involved in the metabolism of various drugs with wide use. Polymorphism at the CYP2D6 locus is one of the most widely known causes for pharmacogenetic variability in humans beings. This review focuses on the importance of CYP2D6 polymorphism in the metabolism of tamoxifen, relationships between the genetic polymorphism and prognosis of patients who have underwent endocrine therapy, and evidences indicating that CYP2D6 may be used as a predictive marker for choosing optimal endocrine therapy for patients with breast cancer.

Objective To analyze the impact of medical insurance reimbursement mechanism on the income of urban community health centers run by various entities. Methods The data of the baseline survey of community health centers in 28 cities made in 2007 were called into play. These data were used to compare the percentage of the annual medical insurance income from outpatient clinics of three types of community health centers in their gross annual income and their annual outpatient clinic income. Results For 929 community health centers with income from medical insurance reimbursement, the average percentage of their annual income from outpatient clinic accounts for 24% of their gross annual income and 26. 09% of their annual outpatient income;In respect of the reimbursement rates of medical insurance at outpatient clinics, the highest come from those run by enterprises, secondly those by the government,and the least those run by social organizations or individuals. Conclusion The mechanism of medical insurance reimbursement has already affected the income of the third type of community health centers. It is recommended to raise the coverage rate of medical insurance of such type of centers and their reimbursement rates of medical insurance. This may further consolidate the policies of medical insurance reimbursement.

Objective To study the effects of changes of neuropeptide Y(NPY) and neurotensin(NT) concentrations in plasma and their ratio imbalance on the incidence of essential hypertension (EH). Methods NPY and NT in plasma in 176 patients with EH were measured with radioimmunoassay(RIA). Results NPY in the EH patients was higher than that in the normal subjects (P

Objective:To obtain anti HCAP1 polyclonal antiserum and define subcellular localization of HCC associated protein(HCAP1).Methods:The HCAP1 was expressed in E.coli DE3 cells and purified.The antiserum was prepared by immunizing rabbit with purified HCAP1 protein.The expression of HCAP1 was analyzed by Western blot in the HCC cells lines.The subcellular localization was observed using fluoroimmunocytochemistry.Results:The rabbit antiserum was obtained and HCAP1 was localized in cytoplasm and nucleoli.Conclusion:The expressed HCAP1 and anti HCAP1 antiserum could be used for studying the HCAP1 role in the HCC;The expression pattern of HCAP1 was mainly in cytoplasm and it was also detected in the nucleoli.

Purpose　To introduce a method to isolate cDNA clones using bacteriophage P1-derived artificial chromosome (PAC) or bacterial artificial chromosome (BAC) as probe for hybridization and try to find some novel genes related to hepatocellular carcinoma.　Methods　PAC 579 (D17S926 locus) and BAC 1529 (D17S1529 locus) in the deletion region of chromosome 17p13.3 in human hepatocellular carcinoma were chosen to screening the human liver cDNA library as probe for hybridization. The isolated positive cDNA clones were partially sequenced, then analyzed by computer comparison and Southern blot.　Results　After three cycles of screening, 78 and 8 candidate positive cDNA clones were isolated using PAC 579 and BAC 1529 probes respectively. Further analysis indicated 18 cDNA clones isolated by PAC 579 probe and 5 cDNA isolated by BAC 1529 probe were potential novel genes related to hepatocellular carcinoma.　Conclusions　The isolation of cDNA clones using PAC and BAC probes is effective and practical.

Objective To investigate the association of promoter hypermethylation of secreted frizzled-related proteins (SFRPs) in patients with colorectal cancer. Methods The promoter hypermethylation of SFRPs in 20 sporadic colorectal cancer tissues and adjacent mucosa were detected by methylation-specific PCR. The amplified DNA was subcloned into the T-A cloning vector and sequenced. Two colorectal cancer cell lines (HCT116 and SW480) were treated with 5-aza-2' deoxycytidine for demethylation. The promoter hypermethylation and protein expression of SFRPs in colorectal cancer cell lines were detected by methylation-specific PCR and Western blotting. Results It was demonstrated that the hypermethylation of SFRP 1, 2, 4 or 5 was 19/20,17/20,3/20 or 13/20in cancer tissues, respectively, whereas it was 12/20, 8/12, 1/20 or 7/20 in adjacent mucosa,respectively. SFRP 1, 2 or 5 methylation was more frequently found in cancer tissue than in adjacent mucosa (P～0.05). Methylation of SFRP 1, 2, 4 and 5 were found in HCT116 cell line, but only SFRP1 and SFRP2 were found in SW480 cell line. There was a negative correlation between protein expression and methylation of SFRPs. The Western blotting revealed that SFRP protein re-expressedafter it treated with 5-aza-2' deoxyeytidine. Conclusion Methylation of SFRP 1, 2 or 5 gene is associated with the evolution of eolorectal cancer, and is closely related to silencing expression.

Objective:To investigate the reversing effect of phosphatidylinositol 3-kinase (PI3K) inhibitors, LY294002(LY) and wortmannin (Wort), on the drug resistance of mitoxantrone (MIT)-resistant human breast cancer MCF-7/MIT cells. Methods:Drug-resistant MCF-7/MIT cells were treated with LY or Wort combined with MIT. Cell viability and proliferation were measured using the MTT assay and morphological changes were recorded by microscopy. Intracellular accumulation of MIT in MCF-7/MIT cells was detected by flow cytometry. Mitochondrial membrane potential was determined by rhodamine 123 staining. Cell cycle was examined by propidium iodide staining. Results:LY significantly enhanced the cytotoxicity of MIT to MCF-7/MIT cells. In LY and MIT cotreated cells, the percentage of cells arrested at S and G2/M phases and the mitochondrial membrane potential decreased significantly compared with single LY- or MIT-treated cells. The mechanism was related with increased accumulation of MIT in MCF-7/MIT cells induced by LY. While Wort, another PI3K inhibitor, did not significantly enhance the cytotoxic effects of MIT.Conclusion: The PI3K inhibitor significantly enhances the sensitivity of MCF-7/MIT cells to MIT.

Objective This is the first study to explore clinical application value of serum Dickkopf-1 (DKK-1) detection in diagnosis of heptocellular carcinoma (HCC) by magnetic solid phase chemiluminescent immunoassay.Methods The level of serum DKK-1 and AFP in 205 cases of HCC,40 cases of liver cirrhosis,and 200 cases of healthy control were quantitatively detected by Magnetic solid phase chemiluminescent immunoassay.The area under ROC curve,sensitivity and specificity of DKK-1 and AFP for diagnosing HCC were calculated.Results The serum level of DKK-1 in HCC group was significantly higher than those of the liver cirrhosis group and healthy control group (P<0.01).DKK-1 maintained diagnostic sensitivity for patients with HCC who were alpha-fetoprotein (AFP) negative (66.3%).ROC curves showed optimum diagnostic cut-off value was 2.4 ng/mL,area under curve (AUC) was 0.822 (95% CI:0.783-0.856),sensitivity 65.9%,and specificity 87.5%).Moreover,measurement of DKK1 and AFP together improved diagnostic accuracy for HCC versus all controls compared with either test alone [AUC 0.915,95%CI:0.886-0.940),sensitivity 81.5 %(P<0.05)].Conclusion Serum DKK-1 detection has an important clinical value for diagnosis of HCC,especially for HCC with AFP negative.The combined detection of serum DKK-1 and AFP can greatly increase sensitivity and accuracy for diagnosing HCC.

Objective: To systematically evaluate the effectiveness of high intensity interval training (HIIT) on body weight and body composition of overweight and obese female college students. In order to provide a theoretical basis for choosing HIIT method. Methods: Randomized controlled trials (RCTs) published till December 14, 2020 were searched in PubMed, the Cochrane Library, Web of Science, Embase, CNKI, CBM, VIP and WanFang Databases. Literature screening was conducted based on inclusion and exclusion criteria, methodological quality evaluation, Meta analysis and publication bias test were conducted on the included literature. Results: There were 20 RCTs, among which 14 studies graded as moderate quality. Meta analysis showed that HIIT could significantly reduce the weight of overweight and obese female college students ( MD =-4.22, 95% CI =-7.20--1.25, P < 0.01 ). Improved body fat rate ( MD =-5.31, 95% CI =-6.88--3.73, P <0.01), BMI ( MD =-2.11, 95% CI =-2.65--1.56, P < 0.01 ), total body fat ( MD =-3.66, 95% CI =-4.89--2.43, P <0.01), abdominal fat ( MD =-0.31, 95% CI =-0.47--0.15, P < 0.01 ), trunk fat ( MD =-2.15, 95% CI =-2.86--1.44, P <0.01) were observed. There was no significant effect on lean body weight ( MD =0.42, 95% CI =-0.94-1.78, P =0.55). Conclusion HIIT can significantly reduce the weight and improve body composition in overweight and obese female college students. It can provide a reliable basis for long term HIIT in overweight and obese female college students.