Objective To analyze the characteristic of serum proteins in non-small cell lung cancer (NSCLC) patients,establish serum markers pattern for the diagnosis of NSCLC. Methods Surface enhanced laser desorption ionization time of flight mass spectormetry(SELDI-TOF-MS) technology was used to analyze serum samples. Biomarker Pattern Software (BPS) was used to detect the protein peaks. Results Sixteen significantly different protein peaks were found in serum samples in NSCLC patients and healthy controls. Eight up-regulated protein peaks and eight down-regulated protein peaks (P

Objective To analyze the epidemud growth factor receptor(EGFR)mutations in NSCLC patients in Fujian province.Methods Fresh specimens of lung cancer and corresponding normal lung tissue were collected from 50 cases of NSCLC patients.After DNA extraction,nested polymerase chain reaction (nested PCR)and direct deoxyribonucleic acid(DNA)sequencing were used to analyze EGFR gene mutations in NSCLC patients.Results EGFR mutations in tumors were identified from 13 of 50(26%)patients,including 10 cages of in-frame deletion in exon 19 and 3 cases of amino acid substitution in exon 21.Conclusion The mjor type of EGFR mutation in NSCLC patients in Fujian is in-frame deletion in exon 19.

Objective To analyze the characteristic of serum proteins in non-small cell lung cancer (NSCLC) patients, establish serum markers pattern for the diagnosis of NSCLC. Methods Surface enhanced laser desorption ionization time of flight mass spectormetry ( SELDI-TOF-MS) technology was used to analyze serum samples. Bio-marker Pattern Software (BPS) was used to detect the protein peaks. Results Sixteen significantly different pro-tein peaks were found in serum samples in NSCLC patients and healthy controls. Eight up-regulated protein peaks and eight down-regulated protein peaks ( P < 0. 001 ) were identified in serum samples of NSCLC patients. Three up-regulated protein peaks(P <0. 05) were identified in serum samples of patients of NSCLC with smoking history. Two up-regulated protein peaks(P <0. 01) were identified in serum samples of patients of squamous carcinoma comparing with adenocarcinoma. No significantly different protein peak was found in serum samples of NSCLC patients at different clinical stages . Conclusion SELDI - TOF - MS technology can identify different protein peaks and so function as a diagnostic tool with high sensitivity and specificity.

Based on the teaching fact and feature of pharmacy specialty. In this article, curriculum location of general microbiology about object, character, function, content design for the higher vocational colleges were disscused. The result would provide some gist to reform teaching methods for microbiology course.

ObjectiveTo clarify the influence of human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein 120 V4 region with mutations at amino acid locuses on its abilities to enter target cells.Methods Based on the facts that ADA strains was a CCR5-tropic strain,only had the ability to infect CCR5 cells; that HXB2 strains was a CXCR4-tropic strain,only had the ability to infect CXCR4 cells,serial glycoprotein 120 mutants with alanine substitution in V4 region of ADA and HXB2 strains,were constructed by overlaping PCR.Eukaryotic expression vectors of mutants and expression vectors of HIV framework gene with luciferase reporter gene were cotransfected into eukaryotic cells to produce pseudoviruse.Concentration of HIV-1 gag P24 in pseudoviruses was detected by enzyme-linked immunosorbent assay(ELISA).U87.CD4.CCR5 and U87.CD4.CXCR4 cells were infected with 20 and 40 ng pseudoviruses,with wild ADA and HXB2 strains as control groups,respectively.The ability to infect cells of pseudovirus of each mutant with HIV-1 V4- region mutated at amine acid locuses 386-417 was measured by detecting the luciferase activity (relative light unit,RLU).ResultsTen mutants with alanine substitution in V4 region of HIV-1 ADA and HXB2 strains were successfully constructed,respectively.Mutants of pseudoviruse with 20 ng and 40 ng at locuses 389-391 and 414-417 with alanine substitution of V4 region in both ADA and HXB2 strains lost completely the abilities to enter CCR5 and CXCR4 expressing cells[ (0 ± 0)％].It was found that introduction of alanine to ADAs 400-403 and ADAs 408-410 increased the ability to infect cells to (124 ± 35)％,(182 ± 29)％ and (127 ± 8)％,( 134 ± 16)％ with pseudoviruse of 20 ng and 40 ng,respectively.Likewise,the ability to infect CXCR4 expressing cells also increased to (144 ± 42 )％ and (121 ± 18 )％ with pseudoviruse of 20 ng and 40 ng,respectively by introduction of alanine to HXB2s 395-397.However,other mutants in V4 region of ADA and HXB2 only maintained partial entry abilities( 15％- 84％).ConclusionsMutants of V4 region of HIV-1 envelope glycoprotein 120 with alanine substitution at locuses 389-391 and 414-417 in both ADA and HXB2 strains have been constructed successfully.They completely lost the ability to enter target cells.

Objective To discuss the influence of the high quality nursing service demonstration project on the doctor-patient relationship.Methods Retrospective study was adopt to compare the nursing service,accompanying rate and patient's satisfaction rate before and after taking the high quality care service demonstration project.Results Nursing service had been improved,with accompanying rate falling from 100.0％to 70.0％ ( x2 =1 116.3,P ＜ 0.05),patients' satisfaction rate increasing from 92.0％ to 99.0％ ( x2 =180.3,P ＜ 0.05 ),the doctors' satisfaction rate increasing from 96.0％ to 100.0％ ( x2 =2.8,P ＜ 0.05 ),and nursing quality control deficiency rate decreasing from 25.0％ to 12.0％ ( x2 =5.6,P ＜ 0.05 ).Conclusions The nursing service demonstration project can ameliorate the doctor-patient relationship by carrying out humanized service.

Objective To investigate the advantages of detection for EGFR gene mutations by denaturing high performance liquid chromatography (DHPLC) technology. Methods DHPLC was used to detect EGFR gene mutations at exon 19 and 21 in 49 cases of non-small cell lung cancer (NSCLC) patients,and the direct DNA sequencing was used to verify the accuracy of DHPLC detection. Results EGFR gene mutation was identified from 13 of 49 cases by DHPLC,including deletion mutation at exon 19 in 10 cases (76.92 %) and alternative mutations at exon 21 in 3 cases (23.08 %). Mutation results of DHPLC was consistent with DNA direct sequencing. The results of the direct DNA sequencing were the same as those of DHPLC. The sensitivity of mutation test by DHPLC was 100 %. Conclusion DHPLC technology can be used for large scale screening of EGFR gene mutation with rapid and accuracy.

Objective To observe the expression of Bcl-2,Bax and proliferating cell nuclear antigen(PCNA) in liver of rats with hepatic fibrosis and the effects of transforming growth factor (TGF)-?1 vaccine on them.Methods Thirty healthy male Sprague-Dawley rats were assigned into 3 groups,named healthy control group(n=10),hepatic fibrosis group(n=10) and TGF-?1 vaccine treated group(n=10).The animal model with hepatic fibrosis was established by injecting solution dimethylnitrosamine (DMN) into abdominal cavity with concentration as 0.5% and dose as 0.2 mL/ 100 g.In TGF-?1 vaccine treated group,every rat was not only injected with DMN but also 150?g TGF-?1 vaccine protein.On the 42nd day,all rats were sacrificed.Then the blood and the liver tis- sues were collected.The expression levels of Bcl-2,Bax and PCNA in liver tissues were detected by S -P immunohistochemistry and observed by routine pathological evaluation.Alanine aminotransferase (ALT),aspartate aminotransferase(AST) and albumin(Alb) were determined by auto biochemical analytical tool.Serum levels of hyaluronic acid (HA),laminin(LN) were detected by radioimmunoas- say (RIA).Results The expression of Bax,which promoted apoptosis,directly correlated with pathological grade in liver of rats,while the expression of Bcl-2 and Bcl-2/Bax,which protected a gainst apoptosis,inversely correlated with pathological grade in liver of rats.The expression levels of TGF-?1 and Bax in healthy control group were significantly lower than those of fibrosis group,how ever,the expression levels of Bcl-2 were comparable between these two groups.As compared with fi- brosis group,the expression of TGF-?1 was significantly lower while the expression of Bcl-2 was sig nificantly higher in TGF-?1 vaccine treated group.However,the expression of Bax was comparable between these two groups.The expression level of PCNA of fibrosis group was significantly higher than that of healthy control group but dramatically lower than that of TGF-?1 vaccine treated group (Both P

OBJECTIVETo investigate the influence of beta-Elemene on expression of ANG II and RhoA/ROCK signaling in Hepatic Stellate Cells.

METHODIn vitro, HSC-T6 cell line was cultured for 24 hours and treated with several concentration of beta-elemene (5.0, 5.0, 2.5 mg x L(-1)) and Y-27632 (30 micromol x L(-1)) for 4, 12 and 24 h. Secretion of ANG II in the supernatant was detected by Radioimmunoassay. The mRNA expression of AGT, RhoA, ROCK-1 and ROCK-2 for 4 h, 12 h, 24 h was detected by RT-PCR respectively.

RESULTOn the time point of 4h, the secretion of ANG II in supernatant by 10 mg x L(-1) beta-elemene was 50.970 +/- 8.081 pmol x L(-1), vs the control group (74.500 +/- 10.999) pmol x L(-1), P < 0.05; 5.0 mg x L(-1) and 2.5 mg x L(-) beta-elemene had no inhibitory effect on the secretion of ANG II, P > 0.05. On the time point of 12h, the secretion of ANG II in supernatant by 10 mg x L(-1), 5 mg x L(-1) beta-elemene was 83.727 +/- 6.850 pmol x L(-1), 91.090 +/- 3.226 pmol x L(-1), respectively, lower than the control (104.367 +/- 5.030 pmol x L(-1)), P < 0.01, P < 0.05. On the time point of 24h, compared with the control (116.620 +/- 7.110) pmol x L(-1)), the secretion ofANGII in supernatant by 2.5 mg x L(-1) and 5 mg x L(-1) beta-elemene was (104.133 +/- 3.296) pmol x L(-1), (100.957 +/- 2.581) pmol x L(-1), respectively, P < 0.05, P < 0.01; but the effect of 10 mg x L(-1) beta-elemene was not obviouse, P > 0.05. Compared with control group, the mRNA expression of AGT by different concentration of beta-elemene were significantly inhibited on different time point (4, 12, 24 h), F value was 30.33, 28.04, 107.19, respectively, P = 0.000. On the time point of 4h, 12 h and 24 h, the RhoAmRNA expression could be inhibited by 2.5, 5.0, 10 mg x L(-1) beta-elemene, (P = 0.000), and there existed no dose dependence. On the time point of 4 h and 24 h, ROCK-1 and ROCK-2mRNA could be inhibited by 2.5, 5.0 and 10 mg x L(-1) beta-elemene, P < 0.01, but on the time point of 12 h, there was no inhibitory effect.

CONCLUSIONBeta-elemene can inhibit the expression of AGTmRNA in HSC and the secretion of ANGII in supernatant. In addition, beta-elemene can inhibit the mRNA expression of RhoA, ROCK-1, ROCK-2mRNA to further regulate the biological effect of ANG II and delay the progress of hepatic fibrosis.

Animals ; Cell Line, Tumor ; Gene Expression ; drug effects ; Hepatic Stellate Cells ; drug effects ; metabolism ; Liver Cirrhosis ; pathology ; RNA, Messenger ; metabolism ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Sesquiterpenes ; pharmacology ; Signal Transduction ; drug effects ; rho-Associated Kinases ; genetics ; metabolism ; rhoA GTP-Binding Protein ; genetics ; metabolism

Objective To establish an easy culture method of successively getting high purity and yield of microglia. Methods Cortices of neonatal Wistar rats (1-3 days old) were employed in this experiment. The first-generation microglial cells were isolated from the mixed glial culture by mechanical means (gently shaking and blowing with pipette). After the mixed glial cells being passaged at a density third generations ofmicroglial cells were harvested. CD1 lb/c, CD45, CD80, CD86 and GFAP were employed as the identification markers in detecting the phenotypes and purity of different generation of microglial cells by scanning electron microscope and flow cytometry. Immunofluorescence staining and CCk8 vitality measurement were used to judge the expression of CD11b/c and detect the proliferation of microglia cells. Microglial phagocytotic function was evaluated by phagocytosis of fluorescent microspheres. Results High yield and purity of microglial cells were stably obtained in this experiment. CD11b/c, CD45, CD80 and CD86 positive expressions were noted in the first and third generations of microglial cells by flow cytometry; CD1 1b/c positive expression was noted in the first,second and third generations of microglial cells by immunofluorescence staining. No obvious differences in the 3 different generations of microglia cells were found on proliferation ability by CCk8 vitality measurement, and on morphology and phenotypes by scanning electron microscope; no obvious differences in the first and third generations of microglia cells were found on phagocytic ability (P＞0.05).Conclusion High yield and purity of microglial cells can successively obtain through the above method;no significant differences are noted among different generations of microglia cells on purity, morphology,phenotypes, proliferation activity and phagocytic ability.