2-Amino-3-hydroxylpyridinE-H2O2-horseradish peroxidase (HRP) voltammetric enzymE-linked immunoassay based on N-heterocyclic substrate has been successfully applied for the detection of carcionembryonic antigen(CEA) in human serum. 2-Amino-3-hydroxylpyridine is oxidized with H2O2 catalyzed by HRP, and the resulting electroactive product produces a sensitive voltammetric peak at the potential of 0.36 V(vs. SCE) in Britton-Robinson (B-R) buffer solution. By this voltammetric peak, free HRP can be measured and immunoassay of HRP label can be developed. The enzymE-catalyzed reaction conditions and voltammetric detection conditions have been optimized, and the electrode procedure of the enzymatic product was investigated. The selected optimum reaction conditions were that the reaction medium was pH 6.0 B-R buffer solution for 10 mL reaction solution containing 1.0 mL of 0.2 mol/L B-R buffer solution, 3.0 mL of 8.0 mmol/L 2-amino-3-hydroxylpyridine solution and 1.5 mL of 0.5 mmol/L H2O2 solution, and the reaction time was 30 min at 37 ℃. The optimum detection conditions were that the supporting electrolyte was pH 7.0 B-R buffer solution for 10 mL of the overall detection solution containing 5 mL of reaction solution and 1.0 mL of 0.2 mol/L B-R buffer solution. The optimum instrumental conditions for the detection were chosen as follows: the initial potential, 0.00 V; the final potential, -0.80 V; the potential scanning rate, 400 mV/s; the mercury drop standing time, 7 s. Under the optimum conditions, the linear range for detection of free HRP was 4.0×10-4-1.0 μg/L with a detection limit of 0.12 μg/L. Based on the new immunoassay system, the linear range of the detection to CEA was 0.50-80 μg/L and the detection limit was 0.50 μg/L, which is 10 times lower than that of traditional spectrophotometric enzymE-linked immunosorbent assay method. The 2-Amino-3-hydroxylpyridinE-H2O2-HRP voltammetric enzymE-linked immunoassay new system exhibits excellent performance having wider linear range and lower detection limit. The new method is inexpensive and simple. It has a great potential in clinical diagnosis.

OBJECTIVETo observe the effect of Astragalus injection (AI) combined with chemotherapy on quality of life (QOF) in patients with advanced non-small cell lung caner (NSCLC).

METHODSSixty-NSCLC patients were randomly divided into the treated group (n = 30, treated with AI combined with chemotherapy) and the control group (n = 30, treated with chemotherapy alone). Chemotherapy of MVP protocol was applied to both groups. AI was supplemented to the treated group by intravenous dripping 60 ml per day. Treatment of 21-28 days as one treatment cycle, and 2-3 treatment cycles were applied.

RESULTSThe effective rate in the treated group was 40.0% and in the control group was 36.7%, the mean remission rate in the treated and control group was 5.4 months and 3.3 months, the median survival period 11 months and 7 months, and the 1-year survival rate 46.75% and 30.0%, respectively, the differences of these indexes between the two groups were all significant (P < 0.05). Moreover, the clinical improving rate and QOF elevation rate in the treated group was 80.4% and 43.3%, as compared with those in the control group (50.0% and 23.3% respectively), the difference was also significant (P < 0.01).

CONCLUSIONAI combined with chemotherapy can significantly improve the QOF in NSCLC patients of advanced stage.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Astragalus membranaceus ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Carcinoma, Squamous Cell ; drug therapy ; Cisplatin ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Mitomycins ; therapeutic use ; Phytotherapy ; Quality of Life ; Vinblastine ; therapeutic use

Aged ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Carcinoma, Merkel Cell ; metabolism ; pathology ; Carcinoma, Small Cell ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Lymphoma ; metabolism ; pathology ; Melanoma ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism

Objective: To determine protein binding characteri stic and signal transmission pathway of melatonin（Mel） receptor(MR) in human e mbryonic peripheral organ tissues. Methods: MR was measured by radio ligand-binding assay and the effect of GTPγS on melatonin specific bindi ng was studied. Results: Mel specific binding sites were det ermined in 16 kinds of human embryonic tissue and this binding could be inhibit ed by GTPγS, supporting the theory that MR is coupled to inhibitory G-proteins system. Conclusion: MR is measured in human embryo tissue, the se results provide experimental data for elucidating the mechanism of the effect of Mel.

0.05).The change of heart rate in 2 groups was greater than that of distilled water respectively(P

Objective To develop a method for detecting anti-cyclic citrullinated peptide (anti-CCP)in the serum of rheumatoid arthritis patients. Methods The range of lineage correlation, precision and detection range of time-resolved fluoroimmunoassay (TRFIA) was analyzed. Thirty-two positive serum of antiCCP, and the sera from 50 health blood donors, 32 SLE patients, 26 patients with SS, 10 patients with scleroderma, 20 patients with MCTD and 18 patients with MS were tested in this study. The clinical specificity was assessed. The consistency between TRFIA and ELISA was analyzed by Mc Nemar test. Results Analyzed by SPSS 13.0 statistical software, the intra-batch precision (n=20) rate was 2%, 3% and 4% respectively, and the inter-batch precision (n=8) was 3%, 4% and 7% respectively for 3 different concentrations. The clinical specificity was 98%, 97%, 96%, 100%, 95% and 100% in the sera of healthy blood donors, SL,E, SS,scleroderma, MCTD and MS patients respectively. The correlation coefficient was 0.989.The average recovery rate was 101%, and the sensitivity was 1 U/ml. When compared with ELISA, the detection range of TRFIA was wider and also with betterstability. Conclusion TRFIA is a stable method with high sensitivity and wide detection range. It can be used to detect anti-CCP antibody. It is important for early diagnosis of RA and monitor the curative effect of RA. And this method has potential to be used in clinical diagnosis.

Objective To explore the diagnosis value of 320-row detector dynamic volume CT in complex congenital heart disease (CCHD)with double outlet right ventricle(DORV).Methods Seventy-eight patients who proveed DORV by surgery in Xinqiao Hospital of Third Military Medical University were reviewed.Thirty-six patients of group A performed 320-row detector dynamic volume CT by using segmental analysis,and were compared with the group B(42 Cases) performed conventional 64-slice CT respectively.Results In 36 cases of DORV confirmed by surgery in group A,MSCT provided accurate qualitative diagnosis in all cases.The accuracy rate of diagnosis of the group B was 90.7％.There was no significant differences compared with the group A (P＞0.05).There were ventricular septal defect in all the 78 cases,pulmonary stenosis in 56 cases,atrial septal defect in 34 cases,pulmonary hypertension in 21 cases,patent ductus arteriosus in 16 cases,coarctation of aorta in 9 cases.Conclusion The 320-row detector dynamic volume CT has important diagnostic value for DORV of the anatomical diagnosis.

Objective To study the effect of lidocaine on the expression of CD11b/CD18 and MPO in lung injury after hemorrhagic shock in rats. Methods Eighty male Wistar rats weighing 230-270 g were randomly divided into 4 groups: sham operation group ( group Ⅰ, n = 8 ) received operation without shock, shock group ( group Ⅱ, n = 8 ) received hemorrhagic shock without resuscitation , normal saline treatment group (group Ⅲ, n = 32 ) received normal saline after shock, lidocaine treatment group ( group Ⅳ, n = 32 ) received lidocaine after shock. The animals were anesthetized with intraperitoneal pentobarbital sodium 40 mg? kg-1 . Hemorrhagic shock was induced by withdrawing blood from right femoral artery at the rate of 2.0 ml?kg-1? min-1 , MAP was maintained at 40 mm Hg for 60 min before resuscitation. Direct arterial MAP and HR were continuously monitored. Group Ⅳwas received lidoacaine at the beginning of resuscitation with a bonus dose 2.0 mg? kg-1 and then followed continuous infusion1.0mg?kg-1?h-1 for 2 h. Group III was received the same dose of normal saline. Flow cytometric analysis was used to assess the expression of CD11b/CD18 on leukocyte and the change of myeloperoxidase (MPO) in the lung was studied at the end of shock (groupⅡ ) and at 2,4, 8, 12 h after resuscitation (group Ⅲ, group Ⅳ) . The rats were sacrificed and the piece of lung was immediately removed for electron microscopic examination. Result In group Ⅲand group Ⅳ ,the expression of CD11b/CD18 on PMNs and MPO in lung were increased significantly compared with Group Ⅰ. Microscopic examination indicated the longer time of reperfusion ,the more serious injury of the lung. And in groupⅣ ,the changes of CD11b/CD18 and MPO were reduced as compared with group Ⅲ (P

Objective To investigate expression of transforming growth factor-?1(TGF-?1)and connective tissue growth factor(CTGF) protein in renal tissues,and detect the levels of urinary TGF-?1 and CTGF in rats with diabetic nephropathy(DN).To observe the influence of losartan on expression of the two protein in renal tissue and excretion in urine.Methods Wistar rats were treated by intravenous injection of streptozotocin after right nephrectomy to induce DN rat model.The DN rats were randomly divided into two groups:DN experimental group and losartan treated group.The expression of TGF-?1 and CTGF in renal tissue were determined with immunohistochemical staining,urinary TGF-?1 and CTGF were assayed by enzyme-linked immunosorbent assay(ELISA) at 6,12 weeks respectively.Results Compared with losartan treated group,urinary protein excretion and the protein expression of TGF-?1 and CTGF significantly increased(P

Reduced radiosensitivity of lung cancer cells represents a pivotal obstacle in clinical oncology. The hypoxia-inducible factor (HIF)-1α plays a crucial role in radiosensitivity, but the detailed mechanisms remain elusive. A relationship has been suggested to exist between hypoxia and autophagy recently. In the current study, we studied the effect of hypoxia-induced autophagy on radioresistance in lung cancer cell lines. A549 and H1299 cells were cultured under normoxia or hypoxia, followed by irradiation at dosage ranging from 0 to 8 Gy. Clonogenic assay was performed to calculate surviving fraction. EGFP-LC3 plasmid was stably transfected into cells to monitor autophagic processes. Western blotting was used to evaluate the protein expression levels of HIF-1α, c-Jun, phosphorylated c-Jun, Beclin 1, LC3 and p62. The mRNA levels of Beclin 1 were detected by qRT-PCR. We found that under hypoxia, both A549 and H1299 cells were radio-resistant compared with normoxia. Hypoxia-induced elevated HIF-1α protein expression preferentially triggered autophagy, accompanied by LC3 induction, EGFP-LC3 puncta and p62 degradation. In the meantime, HIF-1α increased downstream c-Jun phosphorylation, which in turn upregulated Beclin 1 mRNA and protein expression. The upregulation of Beclin 1 expression, instead of HIF-1α, could be blocked by SP600125 (a specific inhibitor of c-Jun NH2-terminal kinase), followed by suppression of autophagy. Under hypoxia, combined treatment of irradiation and chloroquine (a potent autophagy inhibitor) significantly decreased the survival potential of lung cancer cells in vitro and in vivo. In conclusion, hypoxia-induced autophagy through evaluating Beclin1 expression may be considered as a target to reverse the radioresistance in cancer cells.