Objective To clone and construct eukaryotic expressing vector of bcr-abl fusion gene and to express the gene in the mammal COS-7 cell lines. Methods bcr-abl fusion gene was amplified from human chronic myeloid leukemia (CML) K562 cell lines by RT-PCR and the fragment of cDNA was retrieved,purified and cloned into the pEGFP-N3 eukaryotic expressing vector. After the selection of the positive clone and by restriction enzyme analysis and DNA sequencing, the correct plasmid was transfected into COS-7 cell lines and observed the transient expression. Results A 874 bp DNA fragment was amplified by RT-PCR. The sequence analysis showed it was consistent with bcr-abl gene of GeneBank. RT-PCR, Western blotting analysis provided strong evidences that bcr-abl gene was expressed successfully in transfected COS-7 cells.Conclusion The eukaryotic expressing vector of bcr-abl fusion gene was constructed, it will lay the foundation for further study of bcr-abl gene in the diagnosis and treatment of CML.

Objective To investigate IL-18 expression of monocyte cells and the inhibition effect of fluvastation on its expression in patients with acute coronary syndrome(ACS). Methods Isolated monocyte cells from peripheral blood of patients with ACS were incubated with different concentrations of fluvastatin, and IL-18 mRNA level of the monocyte cells was detected by reverse transcription polymerase chain reaction(RT-PCR). Results The monocyte cells of peripheral blood in patients with ACS expressed IL-18 mRNA. IL-18 mRNA decreased from 54 1?9 9 to 50 1?12 6(P

Objective To establish a rapid, specific and sensitive diagnostic technique for the human T. gondii infection with hematopoietic stem cell transplantation and discuss its clinical significance. Me- thods Fifty-six patients subject to hematopoietic stem cell transplantation were detected with ELISA and PCR. Results Among 56 recipients of hematopoietic stem cell transplantation, 7 were positive for T. gondii antigen and 10 were positive for SAG3 gene fragment respectively with the positive rate being 14.3 % and 17.8 % in the ELISA and PCR screening respectively. Twenty healthy people were negative for anti-Toxo antibody.Conclusion PCR is an accurate, relatively rapid, sensitive and specific method for detecting SAG3 gene of T. gondii, and can be considered a valuable additional tool for identification of T. gondii infections.

OBJECTIVE: To investigate the effects of arc-track private lock pedicle orthopedic fixation system Ⅱ (ALPF Ⅱ) on treatment of spinal diseases. METHODS: A total of 86 patients with spinal diseases were treated using self-made ALPF in the Wendeng Orthopaedic Hospital and were included in this study. Of these patients, 14 suffered from cervical spinal stenosis complicated by cervical vertebral destabilization, 29 from thoracolumbar fractures and dislocations, 15 from lumbar spinal stenosis complicated by lumbar vertebral destabilization, 2 from lumbar spondylolisthesis, 8 from spinal tuberculosis, 6 from ankylosing spondylitis, 9 from idiopathic scoliosis, and 3 from congenital scoliosis. According to conditions, different therapeutic regimens were selected. Postoperatively, regular follow-up was performed to observe vertebral healing, intervertebral height, spinal column sequence, and neurological function recovery. RESULTS: All patients were followed up for 9-30 months (average 12 months). The improvement rate of neurological function, spinal mobility, back pain, and melosalgia was 94.1%, 65.9%, 92.1%, and 87.4%, respectively. The postoperative anterior and posterior vertebral heights were apparently recovered, and kyphotic angle was well corrected. No screw, rod loosening or breakage was found. CONCLUSION: Self-made ALPF Ⅱ is an internal fixation method for treatment of spinal diseases. It provides good reduction, reliable curative effects, less complications, and no biocompatibilities.

AIM: To investigate the protective effects and the mechanisms of prostaglandin E1 (PGE1 ) on ischemia/reperfusion (I/R)-induced lung injury after lung transplantation in rats. METHODS: 36 SD rats were randomly divided into 3 groups ( n = 12 in each) : sham operation group ( control group) , lung transplantation (LT)group and PGE1 treatment group. PGE1 was administered to the rats through intra-venous way from 10 min before the operation to the end of the reperfusion. The wet/dry ratio of lung, lung permeability index and neutro-phil percentage were detected in bronchoalveolar lavage fluid (BALE). Superoxide dismutase (SOD) and malond-ialdehyde (MDA) of lung tissue were measured by color-imetry. Serum level of tumor necrosis factor ?(TNF?)was detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: The wet/dry ratio of lung, lungpermeability index and neutrophils percentage in BALF and MDA content of lung tissue in LT group were higher than those in control group( P

BACKGROUND:Breast cancer stem cel s have a greater impact on the occurrence and metastasis of breast cancer. Under simulated tumor microenvironment, we can better analyze the proliferation and differentiation of breast cancer stem cel s. OBJECTIVE:To explore the tumor microenvironment effect on the differentiation of breast cancer stem cel s. METHODS:Breast cancer cel s and MCF-7 cel s were primarily cultured in fibroblast supernatant and serum-free PCM-2 medium, and formation of breast cancer cel s microspheres was observed. Proliferative ability of breast cancer cel s was detected using MTT colorimetry, and the surface markers of breast cancer stem cel s and epithelial-mesenchymal transition markers were measured using immunocytochemistry and RT-PCR methods. RESULTS AND CONCLUSION:The diameter of primary cel microspheres was larger in the serum-free PCM-2 medium than in the fibroblast supernatant, but the culture speed was faster in the fibroblast supernatant than the serum-free PCM-2 medium. At 3 days of primary culture, the expression of ALDH1 in primary cel s was greatly higher in the serum-free PCM-2 medium than in the fibroblast supernatant. However, the expressions of E-cadherin and vimentin were up-regulated in the fibroblast supernatant than in the serum-free PCM-2 medium. In addition, the expressions of E-cadherin and vimentin in MCF-7 cel s cultured in the fibroblast supernatant were up-regulated, while the expressions of ALDH1 and Oct-4 were downregulated. These findings indicate that the tumor environment has some certain effects on the growth and differentiation of breast cancer stem cel s, and some cytokines secreted from fibroblast supernatant can promote the proliferation and differentiation of breast cancer stem cel microspheres to some extent.

Objective To explore the mechanisms of apoptosis induced by arsenic trioxide and amifostine in human acute promyelocytic leukemia cell lines HL-60 in vitro.Methods HL-60 cells were treated with different concentrations of arsenic trioxide alone and combined with amifostine.The inhibitory ratio of the ceils were measured by MTT assay.and the expression of Survivin Was detected by semiquantitate RT-PCR.Results Proliferation of HL-60 cells exposed to arsenic trioxide dwpped down with increasing dose of the dmg and this effect Was significantly hisher when arsenic trioxide Was used in combination with amifostine.Furthermore.there was a more significant decrease in Survivin expression in HL-60 cells treated with arsenic trioxide in combination with amifostine as compared to the cells treated only with arsenic trioxide.Conclusion Arsenic trioxide induced HL-60 cells to undergo apoptosis by downregulating the expression of Survivin. Amifostine enhanced the sensitivity of HL-60 cells to arsenic trioxide by downregulating the expression of Survivin,thus promoting apoptosis effect.

Objective To investigate the value of interphase fluorescence in situ hybridization(FISH)technique and the detection of fusion gene in the diagnosis of acute myeloid leukemia(AML)M2 and M3 Methods FISH was used to detect the AML1/ETO fusion gene and/or PML/RARα fusion gene in incipient cases including 9 AML-M2, 12 AML-M3 and 10 AML undetermined as AML-M2 or AML-M3 primarily diagnosed by routine morphology though bone marrow,cytochemical staining and immunophenotyping,which can help diagnose and guide clinical therapy.Results 4 of 9 AML-M2 cases were AML1/ETO positive.Among 12 AML-M3 cases,10 were PML/RARα positive.1 case was detected AML1/ETO fusion gene.In 10 untonfirmed M3 or M2,3 case8 showed AML1/ETO,5 showed PMIJRARot fusion gene and the rest showed neither of the genes.Conclusion As a new technique of the molecular genetics,FISH is accurate, rapid and efficient.It would be of significance not only at diagnosis of AML,but also for subsequent clinical decision-making.

ObjectiveTo explore the correlation between IL-6-634C/G gene promoter polymorphism and body mass index (BMI),blood sugar (BS),25-hydroxy vitamin D (25-OH-D),and serum lipid levels by investigating in 8-12-year-old Han children in Shanxi province,China.MethodsIn Datong city of Shanxi province,214 8-12-year-old children were enrolled after obtaining informed consent from their parents.The weight and height were measured and the BMI was calculated.BS,serum lipids,and 25-OH-D were determined.IL-6-634C/G polymorphism were detected by polymerase chain reaction restricted fragment length polymorphism.The effects of genotype on BMI,BS,serum lipids,and 25-OH-D were also studied.ResultsThe genotypes of IL-6-634C/G polymorphism in 214 cases were GG ( 15％ ),GC (40％),and CC (45％).The percentages of C and G allele frequencies were 65％ and35％.The genotypes and allele frequencies showed no gender differences ( P ＞ 0.05 ).However,significantly different GG genotypes frequencies were found between overweight and obese children (38.3％) and other children ( normal weight children: 7.3％ ; thin children: 10.9％ ) (x2 =14.715,P =0.006).Multivariate logistic regression analysis showed that IL-6-634C/G polymorphisms and triglyceride were correlated with overweight and obesity (P ＜ 0.05 ).25-OH-D was not correlated with BMI (r =0.075,P =0.528),BS ( r =0.018,P =0.880 ),triglyceride ( r =- 0.097,P =0.417 ),high density lipoprotein cholesterin ( r =0.038,P =0.751 ),and low density lipoprotein cholesterin ( r =- 0.028,P =0.817 ).25-OH-D was not significantly different between overweight and obesity children.The distribution of three genotypes showed no correlation with 25-OH-D deficiency (x2 =0.622,P =0.733 ).ConclusionsIL-6-634C/G polymorphism exists in Han children in Shanxi province.IL-6 gene 634 GG genetype is a risk factor of childhood overweight and obesity,and may affect lipid metabolism.However,it has no direct impact on glucose metabolism.IL-6 gene 634C/G polymorphism and serum 25-OH-D are not relevant.IL-6 gene 634C/G polymorphism is not related to vitamin D deficiency diseases,and may be not related to bone calcium metabolism.25-OH-D is not relevant with BS and blood lipids level,and also is not associated with childhood overweight and obesity.

Objective To investigate the diagnosis and treatment of congenital tracheoesophageal fistula (TEF) in adult patients. Methods The clinical data of 6 adult patients with congenital TEF who had been admitted to Tangdu Hospital from January 1990 to November 2007 were retrospectively analyzed. All patients had the symp-toms of cough when drinking and/or eating fluid and expectoration. The fistulas of the 6 patients were excised, openings were closed with sutures, and the closures were covered with pedicled pleural flaps. Results All patients were diagnosed preoperatively according to the medical history and imaging examination. All the patients were with H-type TEF. The diameters of the fistulas ranged from 0.3 cm to 1.0 cm, and their lengths ranged from 0.2 cm to 1.5 cm. The outcomes and prognoses of the patients were excellent. All patients were followed up for 5 months to 10 years, and no recurrence occured. Conclusions Congenital TEF should be considered when patients are presented with recurrent and chronic cough and pulmonary suppuration. Congenital TEF can be diagnosed accor-ding to the medical history and imaging examination. Surgical procedure for congenital TEF is safe and effective.