Insulinoma-glucagonoma clone 20 can express at least 6 isoforms in human cells.These isoforms can affect cell apoptosis and proliferation through apoptosis related signaling pathway,such as TRAIL or TNF signaling pathways.Furthermore,insulinoma-glucagonoma clone 20,as a GTP-GDP exchange factor,participates in the transportation of nerve synaptophysins.

Objective To investigate the prevalence of virulence genes(ply, pspA, nanA, lytA, psaA) among Streptococcus pneumoniae recently isolated from clinical patients. Methods The 133 strains were isolated from patients in three teaching hospitals in Chongqing from 2006 to 2008. Polymerase chain reaction was used to screen for virulence genes (ply, pspA, nanA, lytA, psaA). Results The positive rate of lytA, psaA, ply, nanA and pspA in 133 clinical isolates were 94.7%, 85.0%, 82.7%, 84. 2% and 60.2%, respectively. The positive rates of the lytA, psaA, ply, nanA and pspA genes in 87 common serotypes isolates was 100%, 87.4%, 86.2%, 89.7%, 67.8%, respectively. Conclusion The total positive rates of five virulence genes in the 133 clinical strains were high. The positive rates of five genes in the com-mon serotypes isolates were higher than those in the no-common serotypes. These genes are important virulence genes of Streptococcus pneumoniae. They could be candidates for protein vaccine of Streptococcus pneumoniae.

Objective To investigate long-term biological variability of serum lipids in Chinese. Methods Serum lipids in a Chinese population with relatively stable life styles were monitored with standardized measurements for 1 year (specimens were taken bimonthly) (23 subjects) or 10～15 years (yearly) (100 subjects). Results The total intra-individual variability (analytical and biological variations combined) of total cholesterol, triglycerides, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol averaged 10%, 28%, 16% and 18%, respectively. Conclusion Biological intra-individual variability is a major source of inaccuracy of cardiovascular risk assessments based on lipid measurements. Measures need to be taken to minimize biological variation when medical decisions are to be made in the treatment of lipid disorders.

Objective To develop an HPLC method for the measurement of n-3 fatty acid index of serum cholesteryl esters.Methods Serum triglycerides were hydrolyzed with ethanolic sodium hydroxide and cholesteryl esters (CEs) were extracted with hexane.The extracted CEs were analyzed by reversed phase HPLC with a UV detection at 205 nm.Cholesteryl eicosapentaenoate and docosahexaenoate ( major n-3 fatty acid cholesteryl esters) were identified by liquid chromatography-tandem mass spectrometry and cholesterol in each CE fraction was measured.Peak areas of CEs were corrected for cholesterol and CE n-3 index was calculated using the corrected peak area and expressed as the percentage of n-3 fatty acid CEs in total CEs.Results The HPLC analysis can be finished in 6 minutes.Triglycerides which interfere with the determination of n-3 fatty acid index, were hydrolyzed with ethanolic sodium hydroxide (4 mol/L) in 30 seconds.The within-run and total CVs for CE n-3 index averaged 0.66% and 0.90%, respectively.CE n-3 indexes of 70 volunteers and 36 coronary heart disease patients apparently healthy subjects and patients with coronary heart disease in Beijing Hospital appeared to be positively skewed and leptokurtic distribution ( skewness = 1.25, kurtosis = 1.70 ).The median of n-3 indices were 0.98% ( 0.37% - 2.40% ).The logarithm of n-3 index appeared to be normal distribution and the average is 0.003 7% with standard deviations of 0.15.The distribution of n-3 indices of gender groups was similar with the total.The medians of females and males were 1.08% (0.60% -2.40%) and 0.95% (0.37% -2.11%) respectively, and the former were significantly higher than the latter( t = - 3.021, P = 0.003 ).Conclusion A new method for the measurement of n-3 index of serum cholesteryl esters by HPLC has been established.It is simple and precise and can be used in predicting cardiovascular diseases risks and monitoring dietary intake of n-3 fatty acids.

Objective To study the expression of thymic stromal lymphopoietin(TSLP) and the activation of DCs in OVA-induced murine asthma model, and investigate the effects and underlying mecha-nisms of TSLP on lung inflammation. Methods Thirty BALB/c mice were randomly divided into control group, OVA group and TSLP neutralizing antibody treated group. The asthma model was evaluated by airway responsiveness and histological analysis of lung tissues ; The levels of TSLP mRNA in lungs were determined by quantitative real-time PCR; The expression of TSLP in lungs were determined by immunohistochemistry and Western blot; The expression of CD40, CD80, CD86 in BALF was detected by FACS. Results Both the histological analysis of lung tissues and the airway responsiveness were all consistent with the characteris-tic of murine asthma model. The expression of TSLP and TSLP mRNA in the OVA group was significantly in-creased compared with blank group. The expression of CD40, CD80, CD86 in BALF from OVA group was increased significantly compared with the control group. Furthermore, treating mice with TSLP neutralizing antibody reduced the expression of CD40, CD80, CD86 on dendritic cells, and IL-4, IL-5, IL-13 in the OVA group. Conclusion Our study indicate that TSLP was highly expressed in the bronchial epithelia of murine asthma model, via upregulation of CD40, CD80, CD86, induce DCs to active CD4~+ T cells and pro-duce type 2 responses, so that aggravating the lung inflammation of asthma. Blocking TSLP is capable of in-hibiting the production of Th2 cytokines, thus presents a promising strategy for the treatment of asthma.

Objective To evaluate the analytical performance of 7 homogeneous HDL-cholesterol reagents. Methods An altracentrifugation-HPLC method was used as the comparison method. Fourty fresh patient samples were analyzed by homogeneous methods and the comparison method. The homogeneous methods were all performed on a Hitachi 7170A chemistry analyzer according to the manufacturer's instructions, Precision, accuracy and total errors were analyzed. Results The homogeneous assays typically demonstrated within run coefficient variance(CV) of < 1%, and total CV of < 3%. Methods A, B and D showed average bias, bias at the medical decision points and total errors all within the NCEP performance criteria and method C and F unacceptable biases (-19.74% and 11.46%, respectively) and total errors according to the NCEP criteria. However, all the homogenous methods (A-F) had total errors of < 30%, as required by the US Clinical Laboratory Improvement Amendment (CLIA). Conclusions Homogeneous HDL-C assays have been shown to be reasonably precise, but discrepant results have been observed with some of the assays. Clinical laboratories should pay more attention on selecting and validating homogeneous HDL-C reagents.

Objective To standardize total cholesterol (TC) and HDL cholesterol (HDLC) analytical systems with the US CDC TC reference method and HDLC designated comparison method (DCM). Methods CDC TC reference method and HDLC DCM were set up and the quality was controlled by participating in the CDC Cholesterol Reference Method laboratory Network (CRMLN) bimonthly survey. The performance of 21 TC or HDLC analytical systems from 3 manufacturers were tested with the methods according to the CRMLN certification protocols. Results The coefficient variation (CV) of TC analyses with the reference method in 18 surveys averaged 0.29% and the bias versus CDC target value 0.1%. The DCM HDLC CV in 17 surveys averaged 0.010 mmol/L(0.39 mg/dl) and the averaged biases versus CDC target and group mean were - 0.019 mmol/L (-0.72 mg/dl) and - 0.006 mmol/L (-0.25 mg/dl), respectively. Most of the TC and HDLC analysis events (> 90%) satisfied the CRMLN accuracy and precision criteria for the reference method and DCM. Eighteen of the 21 tested TC or HDLC systems met the performance criteria for analytical systems and were certified for traceability by CDC. Conclusions A reference method for cholesterol and a DCM for HDLC and performed within an international reference laboratory network have been established and used for certification of TC and HDLC analytical systems, Further application of the methods to the standardization of lipid analysis are expected.

Objective To investigate the angiographic manifestation of the proper esophageal artery (PEA),the hish risk factom for the presence of the anomalous PEA in hemoptysis and to evaluate the safety of transcatheter aaefial embolization(TAE) of the PEA using gelatin sponge(GS).Methods Selective esophageal arteriography WSS performed in forty-three patients with hemoptysis,including 15 cases of pulmonary tuberculosis,18 cases of bmnchiectasis,7 cases of posttuberculous bronchiectasis and three cases of lung cancer. One case experienced failure of bronchial arterial embolization. The angiographic manifestation of the PEAs Was studied.The complications of the procedure and clinical results were observed in the patients who underwent TAE using GS.Results Thirty-nine PEAs were catheterized selectively in 37 patients(86.0%).Eighteen anomalous PEAs(46.2%)were catheterized selectively in 17 patients (45.9%).The anomalous PEAs showed tortuosity,dilatation,hyperplasia,shunting with pulmonary artery and anastomosis with the bronchial artery.All lesions involved basal segment of inferior pulmonary lobar. Bronchiectasis Was the most frequent disease for PEA abnormality. No complications occurred and satisfactory curative effect Was achieved with TAE of the anomalous PEAs.Conclusions It is necessary to perform selective proper esophageal arteriography when the lesion involves basal segment of inferior pulmonary lobar in hemoptysis.Supplemental TAE of the anomalous PEA using GS is safe and valuable in the management of hemoptysis.

Objective To design a new cancer vaccine by using alpha-galactosylceramide (α-Galcer,α-GC) loaded tumor cells in combination with TLR 9 ligand and to evaluate its therapeutic effects on colon canc-er in mice.Methods MC38 cells were transfected with lentivirus (GFP-CD1d) to prepare CD1d-MC38 cells. The expression of CD1d molecules in CD1d-MC38 cells was detected by fluorescence microscopy , RT-PCR and flow cytometry.The sorted CD1d-MC38 cells were loaded with α-Galcer to prepare CD1d-MC38/α-GC complex. Flow cytometry was performed to evaluate the efficiency of combination .A mouse model of colon cancer was es-tablished to investigate the therapeutic effects of α-Galcer loaded tumor cells in combination with TLR 9 ligand ( CD1d-MC38/α-GC+CpG1826) on colon cancer in mice by analyzing tumor growth and mice survival time .Im-munohistochemical staining was used to detect CD 4+T and CD8+T infiltrating lymphocytes in tumor tissues .Re-sults The MC38 cancer cells that expressed CD 1d and GFP were successfully constructed , among which 98.10%±2.53%were positive for CD1d.Moreover, the CD1d-MC38 cells could combine with α-Galcer effec-tively in a dose and time dependent manner .Compared with PBS treated group ,α-GC treated group and TLR9 ligand treated group , the experimental vaccine strategy was sufficient to inhibit the growth of established tumors and prolong survival of tumor-bearing mice (P<0.01).Immunohistochemistry analysis revealed that levels of CD4+T cells and CD8+T cells in experiment group were significantly higher than those in groups treated with PBS,α-GC and TLR9 ligand (P<0.01).Conclusion CD1d-MC38/α-GC in combination with CpG1826 could efficiently inhibit the growth of established tumors and prolong survival of tumor-bearing mice .Immunohisto-chemistry analysis revealed that CD 4+T cells and CD8+T cells played important roles in anti-tumor immunity.