BACKGROUND: Osteosarcoma cell OS-9607 is a cell line cultured from human osteosarcoma tissue. To construct cDNA library of this cell line so as to clone target gene from it.OBJECTIVE: To construct cDNA library of osteosarcoma cell OS-9607 by switching mechanism at 5' end of RNA transcript (SMART) technique and analyze its quality.DESIGN: Verified experimental study SETTING: Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA MATERIALS: This experiment was carried out at the Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA in May 2005. Osteosarcoma cell OS-9607 was cultured in CO2 incubator. Cells were pre-treated with DM for 30 minutes, recovered for 24 hours in standard culture medium. The whole RNA of osteosarcoma cell OS-9607 was isolated by RNAease reagent kit. The quantity and integrity of total RNA was detected by ultraviolet spectrometer and electrophoresis on a denaturing formaldehyde agarose gel stained by ethidium bromide (EtBr). METHODS: Total RNA was extracted from human osteosarcoma cell OS9607 and mRNA was isolated, cDNA of OS-9607 cells was acquired by in situ polymerase chain reaction (PCR) and long-distance PCR (LD-PCR),then the cDNA was ligated with dephosphorylated λ phage vector. The recombinants were constructed with SMART cDNA library construction kit.The size of cDNA inserts and the diversity of library were detected by PCR.MAIN OUTCOME MEASURES: The quality and integrity of total RNA, the titer of un-amplified and amplified library and recombination fraction; analysis of cDNA length of library RESULTS: ①The concentration of the isolated total RNA was determined and the measuring absorbance was between 260 nm and 280 nm. Its absorptance was about 1.9 A. 0.8％ denaturing formaldehyde agarose gel electrophorosis showed clear 28S rRNA and 18S rRNA,and the brightness rate of 28S rRNA and 18S rRNA (28S/18S) was 2:1. ②After linked to λ phage in vitro, packed and transfected host bacteria, the titer of unamplified library was 2.2×107 pfu/mL, and the titer of amplified 1 ibrary arrived to 4.5×1010 pfu/mL. The recombination fraction analysis showed that there were 8 blue plaques and 1 360 colorless plaques in one plate, so the recombination fraction was 99.5％. ③ The obtained cDNA library consisted of 1.6×106 recombinant bacteriophages and the average exogenous inserts of the recombinants was 1.5 kb.CONCLUSION: These results suggest that the obtained osteosarcoma cell OS-9607 cDNA library has high quality, which sets up a firm foundation for the further research on osteosarcoma cell OS-9607 and its elated genes screen.

Objective To explore the relationship between interleukin-8 (IL-8) gene 1633C/T polymorphism and late-onset Alzheimer's disease in order to provide genetic reference data for late-onset Alzheimer's disease.Methods Polymorphism distribution of IL-8 gene 1633C/T in 80 LOAD and 80 normal controls were detected by polymerase chain reaction-restriction fragment length polymorphism(PCRRFLP) analysis.The alleles and gene frequencies type were calculated by the direct counting.Type of gene was proceeded with the Hardy-Weinberg balance estimate.The other statistical analyses were carried out with SPSS13.0 software.The genotype frequencies and allele frequencies were compared with the Chisquare test (x2 test).Results For IL-8 gene1633 C/T,the genotype frequencies (CC,20.0％ ; CT,41.2％ ; TT,38.8％) in the LOAD group were not significantly different from those (CC,11.3％ ; CT,40.0％ ; TT,48.7％) in the control group (P ＞ 0.05).Conclusions IL-8 gene 1633 C/T polymorphism is probably not related to the genetic susceptibility of late-onset Alzheimer's disease.

10.3969/j.issn.2095-4344.2013.25.018

ObjectiveTo explore the effects of personality characteristics and mental health of students on the career orientation.MethodsThe method of stratified random sampling was used in three universities of Jiangxi province.501 subjects of Arts,Science and Medical Science were tested with Holland career orientation test,symptom check list(SCL-90) and 16 personality factors(16PF).Structure validity analysis,analysis of variance and t -value test were employed with LISREL8.70 and SPSS13.0.Results ( 1 ) The proportion of students choosed the type of occupation were:social type 36％,management type 18％,conventional type 17％,reality type 14％,investigation type 9％,artistic type 6％ ; (2) Career orientation,personality characteristics and mental health had moderate effect on the career orientation.The correlation coefficients of personality characteristics and mental health to the vocational orientation were 0.63 and 0.44.The interaction correlation coefficient was 0.56.ConclusionPersonality characteristics and mental health of college students are important factors to the career orientation and decision-making of job seeking.The personality has stronger explanatory power than mental health.Great emphasis should be paid on both short-term psychological health of college students and long-term cultivation of healthy personality in the employment psychological guidance.

ObjectiveTo evaluate characteristic imaging findings of tumor attenuation in 4-phase CT between renal oncocytoma ( RO ) and clear-cell renal ceil carcinoma ( ccRCC ) of small tumor size (≤Scm).Methods Fifty-six patients with histologically confirmed renal masses (11ROs and 45 ccRCCs) were included in this study.Heterogeneous enhancement was found all tumors during the corticomedullary phase (CMP).The CT values of the normal renal cortex,the relatively high enhanced region and the relatively less-enhanced region of the tumor were measured in each phase.Statistical comparison was carried out by Chi-square test or Mann-Whitney test.ResultsIn CMP,the CT value of the relatively high enhanced region in RO [ 163.0 HU ( 141.0—178.0 HU) ] was significantly lower than that in ccRCC [ 194.0 HU ( 166.5—235.0 HU) ; Z = -2.847,P =0.004].Compared CMP with the excretory phase,the attenuation of the relatively highly enhanced region in RO [70.0 HU (41.0—86.0 HU)] were significantly lower than that in ccRCC [87.0 HU (65.0—126.5 HU) ] (Z= -2.032,P =0.042).In the excretory phase,9 of 11 ROs had a further enhancement with its relatively less-enhanced region which was significantly higher than that in ccRCC (21/45 ; x2 = 4.391,P = 0.036).ConclusionsIn CMP,the CT value of the relatively high enhanced region in RO was significantly lower than that in ccRCC.Compared with ccRCC,in the excretory phase,the RO had less attenuation of the relatively highly enhanced region with homogeneous density.

MicroRNAs (miRNAs) are a class of small non-coding RNAs which involve in the regulation after gene transplantation.Researches show that miRNA is closely related to the development and progression of non-small-cell lung cancer (NSCLC).They caused a significantly change in NSCLC cells through downregulate tumor suppressor gene,or act as oncogene or work on some targets which is important in cell signal pathway,in the end it lead to oncogenesis,cell proliferation,cell apoptosis and resistance to chemotherapy or radiotherapy.Therefore,miRNA will be hopefully used in the diagnose and therapy of cancer.

BACKGROUND:Autologous hematopoietic stem celltransplantation is an effective treatment for multiple myeloma. After chemotherapy, autologous hematopoietic stem celltransplantation becomes a standard therapeutic regimen for multipla myeloma, and numerous units and centers have reported that. How to reduce toxic and adverse reactions of the drugs, transplantation-related complications and improvement of long-term survival have been present foci.
OBJECTIVE:To summarize the new progress of autologous hematopoietic stem celltransplantation in the treatment of multiple myeloma.
METHODS:We retrieved PubMed and China National Knowledge Infrastructure database, Vip database, Wanfang database, free medicaljournals.com source for articles published from January 2006 to November 2012 concerning autologous hematopoietic stem celltransplantation in the treatment of multiple myeloma. The key words were“autologous hematopoietic stem celltransplantation, multiple myeloma”. A total of 46 articles were included.
RESULTS AND CONCLUSION: Large-dose chemotherapy combined with autologous hematopoietic stem celltransplantation for multiple myeloma obtained better outcomes compared with the traditional chemotherapy. However, many patients could not relieve after single autologous hematopoietic stem celltransplantation, and the disease recurred final y. Al ogeneic hematopoietic stem celltransplantation was limited by donor source, and treatment-associated fatality rate was high, so its use was confined. Therefore, present new development direction included twice autologous hematopoietic stem celltransplantation, autologous transplantation combined with al ogeneic transplantation with reduced-intensity conditioning regimens as wel as drug on the basis of single autologous hematopoietic stem celltransplantation. Novel drug proteasomes inhibitor and immunomodulator in inducer remission, pretreatment and sustaining stages obviously improved total reaction rate of multiple myeloma therapy and long-term survival.

Objective To study portal vein hemodynamics changes after autoiogous bone marrow stem cell transplantation in patients with liver cirrhosis. Methods The hemodynamic parameters, including portal vein diameter, average portal blood flow velocity and spleen size, were determined by Colour Doppler Ultrasonography in 50 patients after autologous bone marrow stem cell transplantation and all cases were followed for up to 6 months. Results (1) After autologous marrow stem cell transplantation, the portal vein diameter reduced significantly at different timepoints compared to before the treatment ([1.41 ± 0. 15] cm, [1. 38 ± 0. 11]cm,[1.36±0. 17] cm vs. [1. 53 ±0. 18] cm,t = 1. 987,1.994,1. 976, Ps < 0.05), but we found no significant difference in the comparisons among different timepoints (F = 2. 35 ,P >0.05). (2) After autologous marrow stem cell transplantation, the average portal blood flow velocity increased significantly at different timepoints compared to before the treatment ([15. 7 ± 3. 6] cm/s, [16. 1 ± 2.4] cm/s, [15. 9 ± 3.0] cm/s vs.[11.4 ± 3. 3] cm/s ,t = 2. 345, 2.460,2. 381, Ps < 0. 01) , but we found no significant differences in the comparisons among different timepoints (F = 2. 14, P > 0.05) . (3) After autologous marrow stem cell transplantation,the spleen size reduced significantly at different timepoints compared to before the treatment ([4.8±0.3]cm,[4.7±0.6]cm,[4.8±0.5]cm vs. [5. 2 ±0. 7]cm,t =2. 289,2. 390,2.425,Ps <0.01), but we found no significant differences in the comparisons among different timepoints (F = 2. 26, P > 0.05) .Conclusion The autologous bone marrow stem cell transplantation can effectively improve the portal vein blood flow,reduce the spleen size,alleviate portal hypertension in patients with liver cirrhosis.

Objective To study the morphological character of the high endothelial venules(HEVs) and the expression of lymphocyte function-associated antigen-1(LFA-1),platelet endothelium cell adhesion molecular-1(PECAM-1) in the rat mesenteric lymph node,inquire into their morphological representation when lymphocytes pass through the HEVs and the regulative action of LFA-l and PECAM-1. Methods To observe their morphological representation when lymphocyte pass through the HEVs wall and analyze the expression of LFA-1,PECAM-1 with the light microscope,electron microscope and immunohistochemistry method in the rat mesenteric lymph node. Results Lymphocytes contacted the endothelial cells by protuberances or membrane protuberances like welding spot,lymphocytes located in and between the endothelial cells or in the junction space of them in HEVs,there were lymphocytes in the lucid layer between inner and outer layers of basement membranes;LFA-1 mainly expressed at lymphocytes that adhered to endothelial cell in HEVs;PECAM-1 mainly expressed at the endothelial cells and the migrating lymphocytes.Conclusion 1.Lymphocytes contacted the endothelial cells by protuberances,then they got across the endothelial cells into the cellular junction space by the intraendothelial and interendothelial,and at last they emigrated from the basement membranes into the lymph tissue;2.LFA-1 involved in firm adheresion between lymphocytes and the endothelial cells;3.PECAM-1 probably had a relationship with that lymphocytes migrated endothelial cells.

Objective To discuss blood collection tubes with different additives and their effects on the testing results of alcohol concentration in blood sam ples. Methods Blood sam ples from 10 volunteers were collected 2 hours after drinking with seven different types of disposable vacuum blood collection tubes, including ordinary tube without anticoagulant, coagulant tube, separating gel-coagulant tube, sodi-um citrate (1∶4) tube, sodium citrate (1∶9) tube, sodium citrate (9∶1) tube and EDTA-K2 tube. The al-cohol concentrations in these blood sam ples were analyzed by headspace gas chrom atography. Results The concentration testing results of the sam e blood sam ples in different types of tubes were different from one to another. The sequence was as follows:separating gel-coagulant tube>coagulant tube>ordi-nary tube without anticoagulant>EDTA-K2 tube>sodium citrate (1∶9) tube>sodium citrate (1∶4) tube, whereas the results of the sam e blood sam ple in sodium citrate (1∶9) tube and sodium citrate (9∶1) tube showed no obvious difference. Conclusion It is better to collect a suspicious drunk driver’s blood sam-ple using a disposable vacuum blood collection tube, with the EDTA-K2 tube being preferred.