Objective To establish a new non radioimmunoassay for detecting insulin. Methods We put together the techniques of chemiluminescent analysis, flow injection analysis and immunoassay and established a method,flow injection chemiluminescent immunoassay, for insulin determination using sandwhich procedure with the horseradish peroxdase (HRP) as label of antibody and using the detection system of synergistic enhanced chemiluminescence of para phenylpenol and sodium tetraphenylborate with HRP catalysed oxidation of luminol. Results Satisfactory results were obtained in the dynamic range 0.15～60.00 ?U with a relative coefficient of 0 997 2 ( P

Objective The author investigated the clinical features of the the patients with acute pancreatitis and hyperlipaemia (hypertriglyceridaemia),in order to intensify the knowledge of the condition.Methods Forty-three patients with acute pancreatitis were randomly selected and assigned into 2 grups,i.e.,those with hyperlipaemia (11cases) and those with billiary type (32 cases).Results Among the 43 cases,7 were acute necrotizing pancreatitis (5 in hyperlipaemia group).Enumeration correlation demonstrated significant positive correlation between hyperglyceridaemia and the necrotizing type of acute pancreatitis (? 2=6.58,P)or diabetics were significantly higher in hyperglyceridaemia group than in acute billiary pancreatitis group, and so was the life-threatening complications.Conclusion ①Hyperglyceridaemia bears significant positive correlation to the necrotizing type of acute pancreatitis and its life-threatening complications as well.②Diet control,reducing body weight and application of anti-hyperlipaemia drugs will be helpful in prevention and treatment of the relapse of the disease.

Objective: To examine the relationship between parental rearing and adolescent personality traits. Methods: A sample of 637 adolescents was tested by CAPI and EMBU. Results: Neuroticism of girls is higher than that of boys, but interpersonal harmony, face and defensiveness of boys is higher than those of girls. Extraversion, openness and agreeableness of only child is higher than those of other adolescents. Most correlations between parental rearing and Adolescent personality traits are significant. Parental warmth and understanding have positive effect on adolescent extraversion, agreeableness and conscientiousness, and father punishment and mother refusal have negative effect on adolescent agreeableness and interpersonal relationship. Conclusion: Adolescent personality is greatly affected by parental rearing.

OBJECTIVE: To study the general pattern and characteristics of the ADR induced by lidocaine.METHODS: A total of 44 ADR cases caused by lidocaine reported in CHKD during 2002～ 2006 were collected and analyzed statistically. RESULTS: Lidocaine-induced ADR were mostly occurred in local anesthesia and block anesthesia,especially in the first time medication and within ten minutes.ADR could involve multiple organs and systems,and the clinical manifestations were complicated and diversified,which were mainly allergic reactions or even death in severe cases.CONCLUSION: Clinical physicians and pharmacists should be alert to the ADR induced by lidocaine and they should stick to the principle of rational drug use.

Objective:To improve the immunoblotting of immunoprecipitated proteins and decrease the interference of immunoprecipitation antibody in the interaction of endogenous proteins. Methods:Transient transfect cells with fusion protein expression vector containing the targeted S5b gene and the FLAG tag, the transfected cells or untransfected cells were harvested to study the exogenous or endogenous protein interaction. The total cell lysate was immunoprecipitated by specific antibody. Then the eluted immunocomplex was separated by SDS-PAGE, and the TrueBlot antibody or conventional antibody was used as the secondary antibody for immunoblotting detection of S5b and its partner (Rpt1 and Rpt2). Results:Clear immunoblotting bands for S5b, Rpt1 and Rpt2 were obtained. Conclusion:TrueBlot antibody prefers the immunoblot antibody to immunoprecipitation antibody, and decreases the interruption of immunoprecipitation antibody to display clear protein band.

Objective To apply a fluorescent nucleotide dye Sybr Green I in detecting telomerase activity and assess its clinical value in the diagnosis of colorectal carcinoma. Methods Telomerase activity was (measured) by telomeric repeat amplification protocol (TRAP) combined with fluorescent nucleotide dye Sybr Green I in 46 cases of colorectal carcinoma tissue specimens and 19 tumor-adjacent tissue (specimens). Results (Telomeric) repeat amplification product was displayed as clear bands in PAGE stained with Sybr Green I; the positive rate of telomoerase activity in colorectal carcinoma tissue was 89.13%, whereas no telomerase (activity) was observed in 19 tumor-adjacent tissue specimens. There was no (relationship) between telomerase (activity) and the level of cell differentiation, Dukes stages, or metastasis of lymph nodes. Conclusions The method of TRAP combined with Sybr Green I to determine telemerase (activity) is an important method for the diagnosis of colorectal carcinoma.

Objective To construct the double-H1promoters siRNA expression cassettes(SECs) targeted to human telomere retrotranscriptase(hTERT),and investigate the interfering effect of its siRNA on hTERT gene expression in HepG2 cells.Methods SECs were constructed by fusing PCR,based on two different human telomerase hTERT gene fragments.When SEGs transferred into HepG2 cells respectively,the SECs were transcripted to the siRNA.The interfering effect of SECs on the telomerase activity in the cells was assessed by telomeric repeat amplification protocol(TRAP) and PCR-EIA.Results SEGs were successful constracted,and the telomerase activity was significantly inhibited when the HepG2 cells were tranfected with SECs.Conclusions The siRNA SECs display a definite RNA interference effect on the expression of telomerase.This method of SECs preparation can be applied for RNAi research in tumor inhibition.

Based on melamine-Cu conjugate and subsequent interruption of formation of polythymine (Poly T)-templated copper nanoclusters (CuNCs), a novel fluorescent strategy was developed for detection of melamine.The method relied on the principle that the coordination of melamine to copper would be unfavorable for the reduction of Cu2+ in the poly T-CuNCs synthesis process, and thereby resulting in the decrease of fluorescence intensity of CuNCs.By this method, the fluorescence response of CuNCs displayed an linear relationship with melamine concentration in the range from 5 μmol/L to 120 μmol/L.The detection limit was 1.5 μmol/L.Furthermore, the assay was successfully applied in the detection of melamine in milk samples with good recoveries.

Electrophoretic-purified human glycophorin A (GPA) was used to produce its derivatives: (1) glycopeptides were separated and purified from GPA by trypsin digestion; (2) preparation of GPA-antibody and GPA glycopeptide-antibody; (3) preparation of deglycosylated GPA (dGPA); (4) incorporating GPA or dGPA into human RBC membrane lipids to form two kinds of liposomes. The products described above were used to test Plasmodium falciparum FCC1/HN merozoites for their ability to invade human erythrocytes. It was found that GPA-liposomes were able to bind with merozoites and dGPA-liposomes had a negative reaction. GPA, GPA glycopeptide, GPA-antibody, GPA glycopeptide-antibody and GPA-liposome all had the effect to hinder the invasion of merozoites into human erythrocyte while dGPA-liposome had no such an effect.

Objective To investigate the effects of exogenous hydrogen sulfide (H2S) on the mitochondrial function during focal cerebral ischemia and the relationship with mitochondrial damage in rats.Methods Eighty male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 5 groups (n =16 each):sham operation group (group S),cerebral ischemia group (group CI),and high,medium and low dose NaHS groups (NaHS1,NaHS2 and NaHS3 groups).The animals were anesthetized with intraperitoneal chloral hydrate.The focal cerebral ischemia was induced by middle cerebral artery occlusion.Normal saline 1 ml/kg was injected intraperitoneally at 3 h after the model was established.NaHS 2.8,1.4 and 0.7 mg/kg were injected intraperitoneally in NaHS1,NaHS2 and NaHS3 groups,respectively.At 24 h after the model was established,the cerebral infarct volume was determined.The changes in the cerebral infarct volume were observed after administration of NaHS.Cerebral specimens on the ischemic side were obtained for determination of the content of H2S and activity of 3-mercaptopyruvate sulphurtransferase (3MST) in brain tissues.The mitochondria were extracted for determination of the activity and swelling of mitochondrial membrane and changes in the total ATPase,superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and malondialdehyde (MDA) content.Results Compared with group S,the cerebral infarct volume was significantly enlarged in CI,NaHS1,NaHS2 and NaHS3 groups,the content of H2 S in brain tissues and activity of mitochondrial 3MST were decreased in CI,NaHS2 and NaHS3 groups,the activities of mitochondria,SOD and GSH-Px were significantly decreased,and the swelling of mitochondrial membrane and content of MDA were increased in CI and NaHS3 groups,the total ATPase activity was decreased in CI group,and SOD activity was decreased in NaHS1 and NaHS2 groups.Compared with CI group,the cerebral infarct volume was significantly reduced,the content of H2 S in brain tissues and activity of mitochondrial 3MST were increased,the activities of mitochondria,total ATPase,SOD and GSH-Px were increased,the swelling of mitochondrial membrane was reduced,and the content of MDA was decreased in NaHS1 and NaHS2 groups.Conclusion The mechanism by which exogenous H2 S mitigates focal cerebral ischemia is related to enhanced anti-oxidant activity of mitochondria and reduced mitochondrial damage in rats.