Chromatography, High Pressure Liquid ; methods ; Drug Residues ; analysis ; Electrochemistry ; Ethidium ; analysis ; Luminescent Measurements ; methods ; Sensitivity and Specificity

Objective To establish a simple and sensitive method for the determination of serum copper by spectrophotometry.Methods Nitro-PAPS was used as a coloring agent for serum copper in the presence of surfactants Tween-80 and Triton X-100 and the formed complex was measured by spectrophotometry.Results The maximum absorption wavelength of the complex was 570 nm and the molar absorption coefficient was 7.95?10~4 L/(mol?cm).The lineafity of the method was up to 63.0 ?mol/L and the recoveries ranged from 98.6% to 103.1%.The within-run and between-run CVs were 2.1%-3.3% and 2.7%-3.8%.The method(Y)was compared with an AAS method(X)and a correlation of Y=1.01X -0.27(r=0.998 2)was obtained.A reference interval(x~-?2s)determined with this method on 68 individuals was 9.7-24.1 ?moL/L.Conclusions A simple and sensitive method for serum copper has been established.It may used for the analysis of serum copper in clinical laboratories.

OBJECTIVE To investigate enhanced immune function of methionine encephalin (MENK) and its anti-tumor mechanism in CT26 colon cancer mouse model. METHODS 3×106 CT26 cells were implanted subcutaneously in BALB/c mice. Four days after, MENK was peritoneally administrated at the concentration of 20 mg·kg-1 for 14 d. The percentage of MDSCs in bone marrow, spleen, blood, tumor and liver were detected by flow cytometry. Non- esterified fatty acid (NEFA), triglycerides (TG) and total cholesterol (T-CHO) in liver homogenate were tested by a NEFA test kit, a TG test kit and a T- CHO test kit respectively. qRT- PCR and Western blot were used to measure mRNA and protein levels of inflammation-, glycometabolsim- and lipometabolsim-associated indexes in liver. RESULTS MENK decreased percentages of MDSCs in bone marrow, spleen, blood and tumor in colon cancer mice. MENK-treated mice displayed elevated ratio of CD4+T and CD8+T cells in spleen as well as increased T and B lymphocytes proliferation. Meanwhile, MENK also ameliorated liver damage reflected by lower levels of GPT and GOT in serum and reduced risks of cancer- associated index including inflammation, high lipid and high glucose. Furthermore, MENK lowered down the levels of NEFA, TG and T- CHO in liver homogenate. MENK treatment decreased expression of p- STAT3, increased expression of p-AKT, IRS1 and Glut4 at protein level as well as reduced lipogenesis-associated genes and elevated glycolysis-associated genes in liver of tumor bearing mice. Also, abated expression of genes associated with MDSCs generation (M-CSF, GM-CSF, IL-6, IL-1β) and migration (S100A9, KC) was observed within shrunken subcutaneous tumor by MENK intervention. CONCLUSION MENK has the ability to strength immune function against colon cancer by reducing MDSCs and improving liver metabolism.

Background and purpose:Chemotherapy plays an important role in the treatment of breast carcinoma by inhibiting the tumor growth and inducing the apoptosis.MAPK transduction pathway is closely related to proliferation and apoptosis of varieties of tumor cells,inhibition of MAPK pathway may increase the efficiency and decrease the toxicity of chemotherapy.Our study was to investigate the effect of MEK inhibitor PD98059 in response of breast cancer cell lines to Epirubicin.Methods:Human breast cancer cell lines MCF-7 and MCF-7/ADR were used as cell models.Epirubicin(EADM),PD98059(inhibitor of MAPK Kinase-MEK),or EADM+PD98059 was added into the culture medium,the expression of MEK2 and p-ERK were measured by Western blot,the growth of the two cell lines were measured by MTT.Results:ERK activity was elevated in MCF-7 after the treatment of EADM,the cells were more sensitive to EADM if combined with PD98059,while in MCF-7/ADR,ERK activity kept unchanged after EADM treatment,and PD98059 has no effect on the sensitivity of cells to EADM.Conclusion:MAPK signal transduction may be activated in some cells treated by EADM,adding inhibitor of MAPK signal transduction could improve the sensitivity of the cells to EADM.

OBJECTIVETo investigate the indications for non-surgical management of traumatic splenic rupture.

METHODSFrom Jan. 2002 to Jan. 2008, 36 patients with traumatic splenic rupture underwent non-surgical management in the First Affiliated Hospital of Jinan University.

RESULTSOf the 36 cases, 32 were successfully managed without surgical interventions, and 4 converted to open surgery. No death occurred in these patients, nor was delayed splenic rupture identified 1 to 5 years after the treatment.

CONCLUSIONHemodynamically index is an important reference to select the patients, and the degree of splenic rupture, the patient's age and conditions of the hospital should be considered.

Accidents, Traffic ; Adolescent ; Adult ; Aged ; Child ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Splenic Rupture ; physiopathology ; therapy ; Young Adult

OBJECTIVETo measure anatomic data of laminar of C7 and evaluate the possibility of the placement of C7 laminar screw.

METHODSTwenty-two specimens of C7 were measured with an vernier caliper and a goniomete and finded out the anatomic data for prominent vertebra laminar screw placement. The indicators included the thickness of the superior, median and inferior part of C7 laminar (T1, T2, T3), the height of the laminar(H), the distance of the anchor point to the medial wall of C7 pedicle and the lateral wall of the lateral mass (L1, L2) and the angle between the axis of C7 lamina and sagittal plane(alpha).

RESULTSThe mean thickness of superior part of C7 lamina (T1) was (4.30 +/- 0.27) mm, (5.32 +/- 0.27) mm in median part (T2) with 27% (6 cases) of specimens thickness less than 4.50 mm and (4.78 +/- 0.26) mm in inferior part (T3). Both median part and inferior part were thicker than 4.50 mm. The mean height of C7 laminar (H) was (14.83 +/- 0.26) mm. The distance from the laminar anchor point to the medial wall of C7 pedicle (L1) and the lateral wall of the lateral mass (L2) was (26.99 +/- 0.48) mm and (41.60 +/- 1.10) mm respectively. The mean angle between the axis of C7 laminar and sagittal plane(alpha) was (49.3 +/- 1.1) degrees.

CONCLUSIONIt is feasible to place screw in C7 laminar and this technique can be applied in 73% of Chinese. Compared with C7 pedicle screw technique, this technique is simple with little risk to the nerve and vascular structures.

Adult ; Bone Screws ; Feasibility Studies ; Humans ; Internal Fixators ; Spine ; anatomy & histology ; surgery

Five structural important residues of rice nonspecific lipid transfer protein LTP110 were mutated by site-directed mutagenesis. Sequence results showed that they were all mutated successfully. After trying various E. coli expression systems, thioredoxin fusion expression system was found to be a proper system to express wild type and mutant LTP110. cDNA sequences encoding wild type LTP110 and the mutants Y17A, P72L, R46A, D43A, C50A were cloned into two kinds of thioredoxin fusion expression vectors. The expression results were compared. In pTrxFus/GI724 expression system, wild type LTP110 and the mutants Y17A, P72L, R46A could be expressed at low level while D43A and C50A could not be expressed normally; in pET32a(+)/BL21 (DE3) trxB- expression system, wild type LTP110 and all mutant proteins could be expressed very well and the levels were higher than that in pTrxFus/GI724 system. LTP110 fusion protein expressed in pET32a(+) vector was purified and its activity was checked by fluorescence labeled fatty acid. Results indicated that the recombinant LTP110 fusion protein has lipid binding activity. This work provides good basis for the further study.
Amino Acid Sequence ; Carrier Proteins ; genetics ; isolation & purification ; metabolism ; Gene Expression ; Genetic Engineering ; Genetic Vectors ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Oryza ; genetics ; Plant Proteins ; genetics ; isolation & purification ; metabolism ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Thioredoxins ; genetics

PCR is a powerful tool for the amplification of genetic sequences. It has been widely applied in molecular biology. It is generally used to amplify short segments (several hundreds basepairs to several kilobasepairs). It is difficult to amplify a long DNA segment. Based on the sequenced genes, it is known that most intact genes are very long. And intact gene is very important for the gene to express specially and effectively. Long PCR is a very useful tool to amplify intact genes for constructing special expression vectors. We have tried several chemicals to optimize long PCR system and found betaine was the best. Betaine, as an amino acid analogue with small tetraalkylammonium ions, could remarkably improve the amplification of long targets from the plant genome. The suitable concentration of betaine was between 1.0 mol/L and 2.5mol/L. We could effectively amplify a 9 kb DNA segment from maize genome DNA and a 16 kb DNA segment from plasmid. It was shown that different primers and different targets (different GC content) needed different concentrations of betaine. Betaine can reduce or eliminate non-special amplification. In the meantime we tried other additive chemicals, such as DMSO, glycerin, formamide. They were no notable results in long PCR.
Betaine ; pharmacology ; Polymerase Chain Reaction ; methods

OBJECTIVETo analyze the Y chromosome microdeletions in the family of the infertile male and to study on the vertical transmission of Y chromosome microdeletions from father to son.

METHODSThe peripheral blood of infertile patients' family male members was extracted and analyzed with modified multiplex PCR. The infertile family tree was drawn according to the results.

RESULTSTwo cases in twelve investigated families had azoospermia factor (AZFc) microdeletion heredity. The others had no heredity.

CONCLUSIONAZFc microdeletion of the Y chromosome can be transmitted to the male offspring naturally,and the same deletion can result in different phenotypes in different individuals.

Adult ; Aged ; Azoospermia ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Family Health ; Female ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Pedigree ; Young Adult

Animals ; Carcinoma, Hepatocellular ; genetics ; metabolism ; virology ; Genes, p53 ; Hepatitis B ; genetics ; metabolism ; virology ; Hepatitis B virus ; Humans ; Inhibitor of Growth Protein 1 ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; virology ; Loss of Heterozygosity ; Nuclear Proteins ; metabolism ; Point Mutation ; Signal Transduction ; Trans-Activators ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Tumor Suppressor Proteins ; metabolism