At present,the commonly used tumor markers,with low sensitivity and low specificity,can 't achieve the purpose of early diagnosis.However,autoantibodies against tumox-associated antigens have been detected in the asymptomatic stage of cancer and thus can serve as biomarkers for early cancer diagnosis.Moreover,due to autoantibodies are found in sera,they can he screened easily using a noninvasive approach.The discovery of panels of autoantibodies with high sensitivity and specificity will help to improve the diagnostic and prognostic for the patients with cancer.

Objective To evaluate the diagnostic value of the combination of serum DKK1 (Dickkopf-1 )and P53 autoantibodies in patients with esophageal squamous cell carcinoma (ESCC).Methods Serum levels of DKK1 and P53 autoantibodies were measured by enzyme-linked immunosorbent assay (ELISA) for the 1 26 patients with ESCC and 60 normal controls.Receiver operating characteristics (ROC)was used to calculate the diagnostic efficiency.Results The serum levels of DKK1 and P53 autoantibodies were signifi-cantly higher in ESCC than those in normal controls [(673.09 ±343.82)pg/ml vs (362.05 ±1 48.07)pg/ml, Z =6.1 58,P <0.000 1 ;(0.398 ±0.546)vs (0.069 ±0.050),Z =3.832,P <0.000 1 ].ROC curves showed the optimum diagnostic cutoff for serum DKK1 was 588.77 pg/ml,with an area under curve (AUC)of 0.780 (95%CI:0.71 5 ~0.844,61 .9% sensitivity,95.0% specificity).Measurement of P53 autoantibodies demonstrated an AUC of 0.674 (95%CI:0.598 ~0.750,45.3% sensitivity,95.0% specificity).The com-bination of DKK1 and P53 autoantibodies yielded an AUC of 0.843 (95%CI:0.788 ~0.897,73.8% sensitiv-ity,95.0% specificity).In early-stage ESCC,combined detection of DKK1 and P53 autoantibodies improved the diagnostic power,with an AUC of 0.903 (95%CI:0.845 ~0.961 ,81 .0% sensitivity,95.0% specifici-ty).Conclusion Serum DKK1 and P53 autoantibodies can be used as potential diagnostic biomarkers for the ESCC.Combined detection of them might aid the early diagnosis of ESCC.

Objective To explore the diagnostic value of the combined detection of serum Dickkopf-1 (DKK1 )and EB viral capsid antigen immunoglobulin A (VCA-IgA)in patients with nasopharyngeal carcinoma (NPC).Methods Serum levels of DKK1 and VCA-IgA were measured by enzyme-linked immunosorbent assay (ELISA)for the 80 patients with NPC and 65 normal controls.Receiver operating characteristic (ROC) curve was used to calculate the diagnostic value.Results The serum levels [M(QR )]of DKK1 in patients with NPC were significantly higher than those in normal controls [580.773 (429.1 46 )pg/ml vs.31 6.1 74 (252.965)pg/ml],with a significant difference (Z=4.846,P<0.000 1 ).ROC curves showed that the opti-mum diagnostic cutoff for serum DKK1 was 611.981 pg/ml,with an area under curve (AUC)of 0.734 (95%CI:0.654-0.81 5,50.0% sensitivity,96.9% specificity).Measurement of VCA-IgA demonstrated an AUC of 0.71 4 (95%CI:0.631-0.798,47.5% sensitivity,95.4% specificity).The combined detection of DKK1 and VCA-IgA demonstrated an AUC of 0.849 (95%CI:0.783-0.91 4,76.3%sensitivity,95.4%spe-cificity).For patients with early-stage NPC,the detection effect of combined detection of DKK1 and VCA-IgA was much better than that in normal controls,with a significant difference (χ2 =23.784,P <0.001 ). Conclusion Serum DKK1 has potential diagnostic value for NPC.Combined detection of DKK1 and VCA-IgA may aid the early diagnosis of NPC.

Objective To explore the association between blood cadmium levels (BCLs) and clinicopathological characteristics of patients with breast cancer.Methods The clinicopathological characteristics and blood specimens of 186 patients diagnosed with breast cancer were collected between July and December 2009.BCLs were detected by graphite-furnace atomizer absorption spectrophotometer.Mann-Whitney U test and Kruskal-Wallis H test were used to compare the BCLs of patients with different clinical characteristics.Spearman rank correlation analysis was used to evaluate the relationships between BCLs and some indices of clinical characteristics.Results The BCLs was 2.280 (1.579) μg/L.The BCLs were significantly different in patients with different age and body mass index (BMI) (Z =-2.075,P =0.038;x2 =7.429,P =0.023).Also,there were significant differences in different T stages (x2 =10.137,P =0.017),M stages (Z =-2.225,P =0.026),clinical stages (x2 =16.060,P =0.001) and human epidermal growth factor receptor-2 (HER-2) status (Z=-2.072,P=0.038).Excepting for age (r =0.126,P =0.066),BCLs were positively associated with BMI,T stage,M stage,clinical stage and HER-2 status (r=0.159,P =0.030;r =0.171,P=0.020;r =0.166,P =0.044;r=0.154,P =0.040;r =0.152,P =0.038).Conclusion The BCLs are associated with some clinicopathological characteristics of breast cancer,and high blood cadmium burden may promote the development of breast cancer.

Objective To investigate the role of oxidative stress in the epithelial-to-mesenchymal transdifferentiation (EMT) of peritoneal mesothelial cells in rat model of peritoneal fibrosis and the effect of probucol on peritoneal fibrosis. Methods The rat model of peritoneal fibrosis was induced by 4.25% high glucose peritoneal dialysis fluid (PDF). The rats were randomly divided into 4 groups:the control group, the saline group, the peritoneal fibrosis group, and the probucol group. A 4 hour peritoneal equilibration test (PET) was performed 4 weeks later. The peritoneal function and net ultrafiltration (UF) volume were determined. The level of malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in peritoneal tissue were examined. The histology of peritoneal membrane was evaluated by light microscopy. E-cadherin and α-smooth muscle actin (α-SMA) protein expression was evaluated by immunohistochemical method and Western blot.Results The mesothelial cells were detached from peritoneal membrane in peritoneal firbosis rats. Comparing with the control rats, the thickness of visceral peritoneum, the level of MDA, and the-SMA protein expression were increased while the net ultrafiltration volume, the level of GSH-Px and E-cadherin protein expression were decreased in peritoneal firbosis rats. All these changes were reversed in the rats treated with probucol.Conclusion Oxidative stress plays an important role in transdifferentiation of peritoneal mesothelial cell in the peritoneal fibrosis rats. Probucol can improve structure and function of peritoneum, and partially reverse the EMT by reducing the oxidative stress.

Objective To evaluate the nephrotoxicity induced by radiographic contrast media with different osmolality in rats with hypercholesterolemia, and to explore the protective effect of pentoxifylline. Methods Forty-eight healthy SD male rats were randomly divided into normal dietary group (NN, n =8) and high cholesterol supplemented dietary group (H, 4% cholesterol and 1% cholic acid, n=40). At the end of 8th week, the rats with high cholesterol diet were randomly divided into five subgroups (n=8, respectively): high cholesterol diet group(HN), high cholesterol plus iso-osmolar contrast media (iodixanol, IOCM) group (HI), high cholesterol plus low-osmolar contrast media (iohexle, LOCM) group (HL), high cholesterol diet plus high-osmolar contrast media (diatrizoate, HOCM) group (HH) and high cholesterol plus HOCM plus pentoxifylline group (HHP). Forty-eight hours after contrast media injection, the rats were executed and blood samples were prepared to determine total cholesterol, triglyceride, serum creatinine, creatinine clearance(Ccr), fractional excretion of sodium and potassium(FeNa, FeK), and angtension Ⅱ (Ang Ⅱ) levels. The renal injury was assessed by HE staining and TUNEL staining, respectively. The expression of NF-κB protein in the renal tissue was detected by using immunohistocbemical method. Results An increase of cholesterol was observed in all the rats with high cholesterol diet. Scr, FeNa%, FeK% and Ang Ⅱ levels of rats in HH group were obviously higher than those in HL and HI groups respectively. Ccr in HH group [(0.11±0.02) ml·min-1·(100 g)-1] was significantly lower than that in HHP group [(0.43±0.03) ml·min-1·(100 g)-1], HL group [(0.25±0.02) ml·min-1·(100 g)-1] or HI group [(0.27±0.03) ml·min-1,(100 g)-1] (P<0.05). TUNEL staining showed that the pewentage of apoptotic cells in HH group [(89.60±6.40)% ] was higher than that of the other groups [NN (2.40±0.77)%, HN (5.60±1.08)%, HHP (8.91±1.44)%, HL (63.34±11.97)% and HI (61.50±9.40)%]. Immunohistochemistry staining showed that the average gray value of NF-κB positive cells in HH group decreased (P<0.05). There were no significant differences of all indices between HL and HI groups (P0.05). Conclusions Contrast media can cause kidney injuries in the rats with hypercholesterolemia. PTX can protect the renal tissue from nephrotoxicity induced by HOCM in hypercholesterolemia.

Objective:To evaluate the relationship between red blood cell distribution width (RDW) and metabolic syndrome (MS) in patients with impaired glucose tolerance (IGT).Methods:A total of 415 patients with abnormal glucose tolerance were screened by oral glucose tolerance test in Changsha Traditional Chinese Medicine Hospital (Changsha Eighth Hospital) from October 2015 to September 2019. General data were collected and blood routine and biochemical indexes were detected. There were 193 cases in the observation group and 222 cases in the control group. The RDW and other clinical indicators were compared between the two groups, the correlation between RDW and other indicators was analyzed, and the risk factors of metabolic syndrome were analyzed.Results:⑴ The RDW, systolic blood pressure (SBP), diastolic blood pressure (DBP), height (Ht), weight (Wt), waist circumferenc (Wc), triglyceride (TG), cholesterol (CHOL), low density lipoprotein (LDL), creatinine (Cr), uric acid (UA), alanine aminotransferase (ALT), high sensitive C-reactive protein (hs-CRP), body mass index (BMI) of the observation group were significantly higher than those of the control group, while the high density lipoprotein (HDL) was significantly lower than that of the control group, with statistically significant difference ( P<0.05); ⑵ correlation analysis showed that RDW was positively correlated with SBP, DBP, Ht, Wt, Wc, TG, CHOL, Cr, UA, ALT, hs-CRP, BMI, and negatively correlated with HDL ( P<0.05); ⑶ binary logistic regression analysis showed that RDW, Wt, Wc, CHOL, HDL, LDL and hs-CRP were independent risk factors for MS in patients with impaired glucose tolerance. Conclusions:The increase of RDW is a predictor of metabolic syndrome in people with abnormal glucose tolerance, which may provide some reference value for the prevention and treatment of metabolic syndrome.

OBJECTIVE:To establish the HPLC fingerprints for Dangua yangmu cream. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.026% phosphoric(gradient elution)at flow rate of 0.8 ml/min,the detection wavelength was 270 nm,the column temperature was 25 ℃ and the injection volume was 20 μl. The chromatographic peak of aurantio-obtusin was used as reference peak to determine the 10 batches of Dangua yangmu cream,and Similarity Evalua-tion System for Chromatographic Fingerprint of TCM(version 2.0)was conducted to identify common peaks and evaluate similari-ty. RESULTS:There were totally 25 common peaks in the 10 batches of Dangua yangmu cream,and the similarity was not lower than 0.921. The validation results showed the fingerprints of 10 batches of Dangua yangmu cream had good consistency with the ref-erence fingerprints. COMCLUSIONS:The established method is specific and reliable,and can provide basis for the quality evalua-tion and control of Dangua yangmu cream.

【Objective】 To explore a method for the construction of antitumor suicide-gene therapeutic system. 【Methods】 The thymidine kinase gene of herpes simplex virus type 1 (HSV1-tk) was orientationally cloned into the retroviral vector plasmid (pLXSN) by DNA recombinant technique, and then the recombinant plasmid of pLXSN-tk was identified by restriction endonuclease cutting and DNA sequencing. Introduced by PolyFect Transfection reagent, the recombinant plasmid was transfected into the packaging cell line PT67. Screened by G418, a cell line, which could produce virus stably, was obtained. The virus was transfected into human gastric carcinomatous cell strain SGC-7901 and the transfected SGC-7901 was applied to evaluate an anti-tumor effect. 【Results】 The identification with restriction endonuclease cutting and DNA sequencing showed that HSV1-tk was successfully inserted into the recombined plasmid of pLXSN-tk. The stable cloned PT67/tk was obtained by screening with G418 and then its amount was enlarged by culture, the titer of the PT67/tk solution being 4?10~4 cfu/mL. The anti-G418 cloned cell line SGC-7901/tk was got by infecting SGC-7901 with the virus. Anti-tumor experiment showed that GCV had an obvious toxic effect on SGC-7901/tk but had no effect on SGC-7901, indicating recombinant retroviral HSV1-tk expressed HSV1-tk gene which had biological activity. 【Conclusion】 Cloning HSV1-tk into the retroviral vector is effective in obtaining the recombinant retroviral HSV1-tk which can express HSV1-tk gene, and also effective in establishing an antitumor suicide-gene therapeutic system. This research naturally lays a foundation for further studying of Chinese medicines having synergistic anti-tumor action with suicide gene.

Deoxycytidylate (dCMP) deaminase is an enzyme belonged to dCMP cyt deam family. The dCMP deaminase from Streptococcus mutans UA159 was cloned and expressed in E. coli, and purified to homogeneity. The FPLC size exclusion chromatography analysis reveals that the S. mutans dCMP deaminase forms hexamer in solution. The protein was crystallized using hanging drop vapour-diffusion method and diffracted to a resolution of 3.1 (A). The diffraction data were collected at BSRF beamline3W1A. The crystals belong to P213 space group, with unit cell parameters a = b = c = 113.2(A), α =β = γ = 90°. Assuming there are two subunits per asymmetric unit, the Matthews coefficient is 3.6 (A)3 ·Da-1. This is the first crystallization report of the wild-type deoxycytidylate deaminase.