Objective To clone human asparagine synthetase(ASNS)gene,express the MS2- ASNS fusion protein through gene engineering and use the purified target protein to immune BALB/C mice, prepare and identify the monoclonal antibody(McAb),which forms the base for studying mechanism of L- asparaginase used as salvage regimen in midline NK/T cell lymphoma nasal type.Methods ASNS gene fragment was amplified by RT-PCR from HepG2 cell line and constructed into prokaryocytic expression vector.Fusion-protein of MS2-ASNS was expressed and used for immunizing BALB/C mice to prepare McAbs against ASNS.Identified the McAb and detected the expression of ASNS in tumor.Results Part of the ASNS reading frame(NCBI,M27396:179-1 420 bp)was cloned and product length of RT-PCR was 1 263 bp.Molecular weight of MS2-ASNS was about 54 700 Da.Two strains of hybridoma secreting ASNS McAbs were obtained.The subtype of the ASNS McAb was IgG2a.Western-blot showed that the McAbs could specifically react with MS2-ASNS fusion protein and ASNS protein in tumor cell lines.ASNS expression was detected by immunocytochemistry and Immunohistochemisrry.Conclusion We have cloned human ASNS gene,obtained the anti-ASNS McAb and examined the expression of ASNS in tumor.

Objective To explore the clinical application and outlook of anterior segment observation with Stratus optical coherence tomography(Stratus OCT).Design Prospective case series.Participants 56 eyes in 38 patients with primary angle closure glaucoma (PACG).Methods Morphological changes of anterior chamber angle (ACA) were observed with Stratus OCT before and 4 weeks after laser iridectomy.Opening status of ACA were described and measured with Photoshop software.Main Outcome Measures Opening degree of ACA and ratio of anterior chamber depth to thickness of cornea.Results To a great deal of extent,Stratus OCT could be used to observe the appearance of ACA. Clear images of ACA could be acquired.The ACA before laser was (15.67?5.33) degree and significantly reopened to(26.56?8.17) degree after laser iridectomy (P=0.000).The ratio of anterior chamber depth to thickness of cornea was also changed from (0.39?0.13) pre-treatment to (0.89?0.32) pest-treatment(P=0.000).Conclusions Stratus OCT could be applied to observe the changes of opening degree of ACA easily.The standard of quantization should be consummated in the future.

Objective To evaluate the clinical application of sweep pattern visual evoked potential in assessing visual acuity (SPVEP-A)of amblyopic children.Design Prospective clinical study.Participants 80 patients(148 case)of amblyopic children and 26 children(52 case)with normal vision.Methods The correlation were analysed of SPVEP-A and E acuity in 80 amblyopic children and of SPVEP-A and PVEP-A in 26 children of normal visual.SPVEP-A was recorded with UTAS-E 3000(LKC)using gratings of 25 differ- ent spatial frequency from 0.2 to 18.8cpd as stimulus.The response were averaged and DFT on the monitor display.SPVEP-A was de- termined by extrapolate to 0 response amplitude.Both transient PVEP-A and SPVEP-A were examined in 26 normal children.The ob- jective visual acuity in PVEP were determined with the highest spatial frequency which evoked a recognizable response.Main Out- come Measures SPVEP-A,PVEP-A and E acuity.Results SPVEP-A were from 0.35 to 0.9 among 80 patients with corrected E acu- ity from 0.1 to 0.8.The correlation coefficient of SVEP-A-I with E acuity was 0.602 .The correlation coefficient of SPVEP-A.between reproducibility detection was 0.448.The correlation between SPVEP-A and E acuity in amblyopic slightly was higher than middle and severe amblyopia(r=0.773 vs r=0.590).In 26 normal children a correlation coefficient of 0.679 was obtained between E acuity anti PVEP-A but 0.424 between E acuity and SPVEP-A.Higher evaluation or lower evaluation of E acuity was found in SPVEP-A in this study.Acuity was upostimated in lower scope of E acuity and underestimated in higher scope of E acuity.The reproducibility and vari- ation were not satisified.Conclusion The sweep VEP is a objective method of amblyopic screening.There is significant positive corre- lation between SPVEP-A and E,but its variability is more great,so we should improve its stability and positive rate uheriorly.

Animals ; Animals, Newborn ; Female ; Hyperbaric Oxygenation ; Hypoxia-Ischemia, Brain ; physiopathology ; therapy ; Nerve Growth Factor ; pharmacology ; Nerve Regeneration ; drug effects ; physiology ; Rats ; Rats, Wistar

The diagnosis and treatment of prostate cancer are being improved due to the popularized screening of prostate specific antigen. Advanced prostate cancer, in spite of its response to androgen deprivation therapy, may finally develop into castration-resistant prostate cancer (CRPC) and shorten the overall survival of the patients. Many efforts have been made by worldwide researchers for new approaches to the management of CRPC, including new hormonal therapy, cytotoxic chemotherapy, immunotherapy, and bone metastasis-targeted therapy. This paper reviews the emerging agents undergoing clinical evaluation and drugs that have received approval for the treatment of CRPC in order to provide doctors and patients with more treatment options for CRPC and improve the overall survival rate and quality of life of the patients.
Androgen Antagonists ; Bone Neoplasms ; prevention & control ; Humans ; Immunotherapy ; Male ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms, Castration-Resistant ; therapy ; Quality of Life

Sulfate-reducing bacteria (SRB) are widespread in the environment. SRB are obligate anaerobes and capable of dissimilatory reduction of sulfate. SRB have application prospects in the control of environmental pollution due to that many pollutants can be removed by SRB. The biological characteristics and metabolic mechanisms of SRB are introduced, and the application of SRB in the treatment of environmental pollution is described in this paper. The research progress of Cr(Ⅵ ) reduction and Cr(Ⅵ ) removal from wastewater by SRB is reviewed, and future direction of research on the control of Cr(Ⅵ ) pollution by SRB is also analysed.

BACKGROUND:Large-scale expansion of undifferentiated and multipotential adipose-derived stem cells using serum-free culture system is a difficult issue to be resolved. OBJECTIVE:To establish an in vitro culture system combined with the extracellular matrix in order to investigate the efficiency, effectiveness and security of extracellular matrix on expanding adipose-derived stem cells. METHODS:In vitro isolated adipose-derived stem cells were seeded in traditional two-dimensional plastic plates and extracellular matrix-coated plates supplemented with serum-free medium respectively. After in vitro expansion, total cellnumber, expression of cellsurface markers, cellsenescence degree and multipotent differentiation ability (adipogenic, osteoblastic and chondrogenic differentiation) of adipose-derived stem cells cultured under both conditions were detected and compared. Moreover, the clinical safety of adipose-derived stem cells expanded in extracellular matrix-coated plates was investigated. RESULTS AND CONCLUSION:Total cellnumber of passage 5 adipose-derived stem cells cultured in extracellular matrix-coated plates was 10 times more than that in traditional two-dimensional plastic plates. Flow-cytometric analysis showed that adipose-derived stem cells cultured with extracellular matrix expressed stem cellsurface markers. cellular senescence examination showed that almost al of passage 15 adipose-derived stem cells cultured with extracellular matrix showed no aging, while most passage 5 adipose-derived stem cells cultured by the two-dimensional system aged and lost their proliferation ability. Multidirectional induction of adipose-derived stem cells showed that passage 15 adipose-derived stem cells cultured with extracellular matrix could stil differentiate into adipocytes, osteoblasts and chondrocytes as passage 5 adipose-derived stem cells did, which performed much better than the induced differentiations of passage 5 adipose-derived stem cells cultured by the two-dimensional system. Karyotype analysis and in vivo invasion experiment insured the clinical safety of adipose-derived stem cells expanded with extracellular matrix. Al above results suggest a safe and more efficient expansion system of extracellular matrix for clinical application using the serum-free culture system combined with extracellular matrix.

ObjectiveTo establish a new method for preparing synaptosomes.MethodsDensity gradient centrifugation method was used to isolate synaptosomes of mouse, checking by transmission electron microscopy.ResultsSynaptosomes prepared by this method had intact morphological characteristics, surrounding with a continuous oval-shaped membrane structure, moreover, mitochondrion and lots of synaptic vesicle in them.ConclusionThis method is applicable to establish a rapid, convenient and useful method for preparing synaptosomes.

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Objective To compare the heart function and observe the clinical prognosis of dilated cardiomyopathy(DCM) in children treated with combination of prednisone and azathioprine and high dose intravenous immunoglobulin(IVIG).Methods From Jan.1998 to May 2004,20 children who met the diagnostic criteria of DCM were divided into 2 therapeutic groups(groupⅠ,groupⅡ) according to the time of hospitalization.On the basis of traditional congestive heart failure(CHF) therapy,the children in groupⅠwere treated with immunosuppressive therapy(prednisone plus azathioprine) and the children in groupⅡ were treated with high dose IVIG.The course of treatment was 6 months for all children.Heart functions in all children were evaluated and echocardiographic parameters including left auricular diameter(LA),left ventricular end diastolic diameter(LV),left ventricular ejection fraction(LVEF) and cardiac index(CI) were measured before and after therapy,respectively.All children were followed up for 1 year to observe survival rate(%).The findings were analyzed by t test and ?~2 test.Results The percentage of heart function increased from Ⅲ-Ⅳ toⅠ-Ⅱin groupⅠ,which was lower than that in groupⅡ(P