Objective:To construct human single-chain variable fragment (scFv) antibodies gene library associated with lung adenocarcinoma. Methods:Total RNA was extracted from the lymphatic tissue near the lung adenocarcinoma.VH and VL fragments were amplified with RT-PCR,the purified VH and VL fragments were used to produce scFv fragment by splicing-overlap-extension PCR. The scFv gene was cloned in pCANTAB-5E Phasmid,then transferred to Ecoli TG1.The insertion ratio of antibody gene was identified with RT-PCR,the positive enzyme-cutting products were identified with 1.5% agarose gel electrophoresis. Results:Clear 28 S and 18 S strips were found in total RNA;The size of VH is about 370 bp,VL is about 350 bp.The size of scFv is about 750 bp.After transformation into Ecoli TG1,2.2?107 CFU/?g ampicillin resistant bacteria colonies grow after overnight culture,and the positive insert ratio was 83.3%. Conclusion:Human single-chain variable fragment(scFv) antibodies gene associated with lung adenocarcinoma was constructed successfully. It could provide bases to further screen the antibody library construction.

Objective:Retrospectively evaluate the clinical value of 99mTc-Sc lymphoscintigraphy in sentinel lymph node(SLN)location in breast cancer,and to offer accurate orientation for sentinel lymph node biopsy(SLNB).Methods:Fifty breast cancer patients underwent 99mTc-SC lymphoscintigraphy,the outcomes were compared with?detection probe.Results:SLN was found in 45 of 50 cases(90.00%),the number of SLN was 1.60 in every case averagely.41 patients were found in concordance with?detection probe,4 cases were not detected by?probe.The accuracy,specificity,false negative rate and false positive rate of SLN to predict axillary lymph node status were 93.33%,100%,6.67%,0%.Conclusion:99mTc-SC lymphoscintigraphy can exactly predict axillary lymph node status and guide?detection probe and SLNB.

Medical education for the international medical students is an important part of home medical colleges. This article focuses on the training of teacher’s ability,the understand- ing of the international medical students,the preparation for lessons and the agile use of many teaching ways during the teaching process of Nuclear Medicine for the international medical stu-dents. Up to now,we still have no textbooks of Nuclear Medicine for the international medical students. This is the problem that exists in the Nuclear Medicine teaching.

OBJECTIVE:To observe the correlation of concentration-time curves of riboflavin in salive and plasma.METHOD:After peple orally taking riboflavin tablets,the riboflavin in salive was determinated by fluorescence spectrophotometry with excitation wavelength 475.4nm and emission wavelength 524.7nm.RESULTS:The recovery of riboflavin was over 96%.The detection limits of riboflavin was 5ng.ml-1,and the calibration curve was linear in the range of 5ng.ml-1～3μg.ml-1.The regression equation was △F′=0.116c+0.232(r=0.9999).CONCLUSION:The concentration-time curves of riboflavin in salive was similar to these in plasma.

Objective To investigate the tumor-inhibitory effect of cytokine-induced killer cells(CIK)co-cul-tured with dendritic cells (DC)pulsed by breast cancer stem cell antigen on the same tumor-bearing mice. Methods Breast cancer stem cells were isolated from the cell line of MCF-7/ADR and extract lyses antigen of the stem cell was saved. DC and CIK derived from peripheral blood mononuclear cells of healthy individuals were co-cultured and pulsed or un-pulsed by the above antigen lyses. This DC+CIK were injected to breast tumorbearing mice (BCSC-AP-DC+CIK group), and were used to compared with the common breast cancer cell antigen (rather than breast cancer stem cell antigen) pulsed DC+CIK group(AP-DC+CIK group), DC+CIK group, CIK CIK group and normal saline group(NS group). The tumor-inhibitory effect were evaluated and compared among all 5 groups through the tumor size, TdT-mediated dUTP nick end labeling test (TUNEL), examining expression level of bcl-2 and bax by immunohistochemistry. Results The tumor size in each group before and after therapy and the tumor size after therapy between each group was of significant difference(P＜0.05). The maximum size is NS group(3.625±0.093)cm3 and BCSC-AP-DC+CIK group is minimum,which is (1.234±0.131)cm3. BC-SC-AP-DC+CIK group is of highest expression of bax and apoptotic index value, lowest bcl-2 expression in all 5 groups. Conclusion The CIK co-cultured with DC pulsed breast cancer stem cell antigen was more effective to induce apoptosis of breast cancer cells than those of CIK cells co-cultured with DC pulsed breast cancer cell antigen,CIK cells co-cultured with DC and CIK cells.

Objective To construct human single-chain antibody gene library associated with lung adenocarcinoma, and screen lung adenocarcinoma cell A549-specific antibody. Methods The lymphatic tissue near lung adenocarcinoma was used to construct human single-chain antibody gene library. The specific antibody to lung adenocarcinoma cell A549 was screened from the antibody library. Positive clone bacteria were transformed to E. coli HB2151, and their dissolvability was observed. The soluble scFv was purified by affinity chromatography and its relative molecular mass was determined by SDS-PAGE and Western blotting. The specificity of scFv to human lung adenocarcinoma cells was identified with ELISA. Results The phage antibody library of 4.6?107 was constructed successfully. Single-chain antibody of human lung adenocarcinoma was enriched. The ratio of yield was increased gradually, 181 times higher after 5 rounds of panning than after the first round of panning. SDS-PAGE and Western blotting results showed soluble scFv' molecular mass was about 30 000. ELISA showed dissolved antibody had high specificity to bind A549 cells, not MDA-MB-435. Conclusion The single-chain antibody gene library of human lung adenocarcinoma was constructed successfully, and specific antibodies to lung adenocarcinoma were screened, providing the basis for single-chain antibody radionuclide imaging and treatment.

Background Optic neuritis is one of the common clinical neuro-ophthalmic diseases.Recurrent optic neuritis can cause irreversible axonal damage and visual impairment,and no prevention method is effective up to now.Its pathogenesis is considered to be associated with the body's immune imbalance.Objective This proposal tried to establish experimental autoimmune encephalomyelitis (EAE) and explore the immunomodulatory effects of T helper (Treg),Th17 cell in the model of optic neuritis.Make the immune mechanism of demyelinating optic neuritis clear.Methods EAE model was established to observe the clinical symptoms and visual electrophysiological changes of optical neuritis in mice;hematoxylin-eosin staining to observe the histological changes in the mouse models of EAE; immunohistochemical staining to detect optic nerve axon damage markers β-amyloid peptide precurso (β-APP) protein expression quantity changes;reverse transcription PCR (RT-PCR) to detect the dynamic expression of forkhead/winged helix transcription factor p3 (Foxp3),transforming growth factor-β (TGF-β),interleukin-1β (IL-1β),IL-17,IL-10 in optic nerve tissue.Results After immunization 11 days,neurological symptoms of EAE mice started to appear,hematoxylin-eosin staining showed optic nerve tissue infiltration of inflammatory cytokines; immunohistochemical staining showed enhancement of axonal injury markers β-APP positive staining; flash visual evoked potential(F-VEP) test showed 7,11,14,19,23,28 days,compared with the normal mice,P1 prepatents of model group were prolonged after modeling,the differences were statistically significant (t =4.487,15.203,16.364,11.540,11.959,16.163,all at P＜0.05).When 7 days after modeling,the difference of N1-P1 amplitude between normal group and model group was no statistical significance (t =-0.992,P =0.378); while 11,14,19,23,28 days after modeling,compared with normal mice,N1-P1 amplitudes of model mice were significantly lower,the differences were statistically significant(all at P＜ 0.05).The expressions of TGF-β,IL-1β,IL-17,Foxp3,IL-10 mRNA among different time points had stastistie significant differences (F =12.721,15.015,14.343,69.374,68.290,all at P =0.000),compared with the normal group,when 11 days,14 days after modeling,TGF-β mRNA levels were significantly increased ; 19 days,23 days after modeling,IL-1β mRNA levels were significantly increased;7 days,11 days after modeling,IL-17 mRNA levels were significantly increased ;7,11,14,19,23,28 days after modeling,Foxp3 mRNA levels were significantly lower; 19,23,28 days after modeling,IL-10 mRNA levels were significantly lower,the difference were statistically significant (all at P＜ 0.05).Conclusions Th17 cells launch demyelinating optic neuritis immune injury,and Th1 subgroup performs the function of maintaining inflammatory lesions,Treg cell subgroup is abnormal much earlier than that of the Th2 subgroup.The imbalance of Th17/Treg cell may be involved in the pathogenesis of demyelinating optic neuritis.

BACKGROUND: Both pulse pressure (PP) and aortic stiffness are the predictors of coronary artery disease (CAD). The relationship between aortic PP and atherosclerosis may be bi-directional. Some investigations have tested the relationship between coronary arteriongraphy-diagnosed coronary heart disease and aorta pulse pressure (APP).OBJECTIVE: To observe the relationship between coronary atherosclerosis and APP in patients with primary hypertension.DESIGN: Case control observation.SETTING: Department of Cardiology, Xuanwu Hospital, Capital University of Medical SciencesPARTICIPANTS: A total of 300 patients with primary hypertension who received the treatment in the Department of Cardiology, Xuanwu Hospital, Capital University of Medical Science between July 2002 and January 2005. The patients were all untreated hypertensive patients who were referred for a first diagnostic coronary angiography. All of them met the diagnostic criteria of international hypertensive association. Secondary hypertension, myocardial disease, valvular disease of the heart (VDH), heart failure, liver and renal insufficiency and so on were excluded. The patients, 170 male and 130 female, were aged (61±11 )years. Informed consents were obtained from all the patients.METHODS: A total of 300 untreated hypertensive patients were recruited for a first diagnostic coronary angiography. According to whether having coronary artery disease (CAD), the patients were divided into CAD group (143 in total, 92 male, 64%) and non-CAD group (157 in total, 82 male, 52%). Informed consents were obtained from all the patients.The following data were collected: invasive intra-aortic systolic blood pressure (ASBP) and intra-aortic diastolic blood pressure (ADBP), extent of coronary artery disease, and basic clinical materials of the patients. All the observed index were expressed as Mean±SD. Independent sample t test was used in the comparison between two groups, and P ＜ 0.05was set as significant difference.MAIN OUTCOME MEASURES: ASBP, ADBP and BP, ratio of stroke volume (SV), which reflects aortic stiffness, to aortic pulse pressure (APP); The number of branch and stegnotic extent of coronary artery disease (Cases with stegnotic extent ＜ 50% were included in non-CAD group; Basic clinical indexes and routine biochemical indexes of patients after admission (including fasting blood glucose, serum creatinine, blood lipid and so on).RESULTS: The enrolled 300 patients with hypertension all participated in the result analysis. In the whole population, ASBP and PP were significantly higher in CAD group than non-CAD group [(150.3±26.5) vs. (145.6±23.3) mm Hg, P ＜ 0.05;(77.1±22.7) vs. (70.4±19.3) mm Hg, P ＜ 0.05)]. The ratio of APP to SV in CAD group was markedly higher than that in non-CAD group(1.20±0.44) vs. (0.96±0.33), P ＜ 0.05]. Fasting blood glucose of patients in the CAD group was significantly higher than that in the non-CAD group [(1.38±0.27) vs. (1.08±0.28) mmol/L, P ＜ 0.01]. Serum creatinine of patients in the CAD group was significantly higher than that of non-CAD group [(11.98±2.15) vs. (11.19±1.58) μ mol/L, P ＜0.01]. High-density lipoprotein cholesterol of patients in the CAD group was significantly lower than that in the non-CAD group [(0.54±0.13) vs. (0.62±0.18) mmol/L, P＜ 0.01].CONCLUSION: Atherosclerosis may further aggravate the aortic stiffness and then cause the increase of APP in patients with primary hypertension.

OBJECTIVE To explore the effective methods for medical personnel preventing and controlling the infection of HIV after occupational exposure. METHODS The 7 cases contacted the AIDS patients had been analyzed and a set of institutions and monitoring methods were established. RESULTS Because of the emergency measures,the 7 medical personnels hadn′t infected HIV for the 2 follow-up years. CONCLUSIONS To think highly of the safety education of occupational exposure and enhance the monitoring and management of HIV infection can decrease the occupational exposure rate and ensure the occupational safety of medical personnels.

Objective The aim of this study was to assess the distribution histogram of arm circumference (AC) in adult hypertensives in Beijing area, and to establish which size of cuffs is most appropriate in clinical practice. Methods We conducted a cross-sectional survey in a cohort of 424 (male 61.6%) consecutive hypertensives patients. Arm circumference was measured at the mid-point of the right upper arm. Subjects were stratified in three groups: group 1 with AC between 22-26 cm (small size), group 2 AC between 27-34 cm(medium size), and group 3 AC great than 34 cm (large size). Using AC of 32 cm as cutoff point, patients were subdivided as group A (AC