Objective To investigate the influences of drop components on the"coffee ring" test of DNA. Methods The DNA-EB drops containing SDS, NaCl, etc, were evaporated on glass slides. After evaporation, the fluorescent Deposition Pattern (DP) and Integrated Optical Density (IOD) was acquired with a gel imaging system. The dose-effect relationship was analyzed. Result When the non-DNA components concentration is low, the DP was still characteristiced by peripheral rings, subtle component-specific differences, such as tree-ring like structure and radial crack, existed. At high concentrations, DP was various, which may be ring + scattered dots (NaCl), central spot + small weak ring (SDS), concentric/tree-ring (TritonX-100) or ring + spot (H+), et al. Non-DNA components had little effect on IOD(0.5~2 times). Conclusion Non-DNA components in DNA-EB drops influences both the DP and IOD, but rough estimation of DNA concentrations is still effective. Moreover, analyzing the DP can provide more informations about sample purity and residue.

OBJECTIVETo construct a subtracted cDNA library of differentially expressed genes in eosinophils from asthma patients.

METHODSSuppression subtractive hybridization (SSH) was used to isolate the cDNA fragments of differentially expressed genes in the eosinophils of asthma patients before and after treatment. The cDNA fragments were directly inserted into T/A cloning vector to establish the subtractive library, followed by amplification of the library through E. coli transformation with calcium chloride and screening of blue and white clones of the transformants. One hundred positive bacterial clones were randomly picked and identified by colony PCR.

RESULTSThe amplified library contained more than 3,000 positive bacterial clones. Analysis of the randomly selected 100 white clones by PCR showed that 90% of the clones contained 100-500 bp inserts, which might be the cDNA fragments of differentially expressed genes in eosinophils of asthma patients before treatment.

CONCLUSIONA subtracted cDNA library of differentially expressed genes in the eosinophils of asthma patients before and after treatment is constructed successfully by SSH and T/A cloning techniques, which lays a solid foundation for screening and cloning new specific differentially.expressed genes in the eosinophils of asthma patients.

Asthma ; blood ; genetics ; DNA, Complementary ; genetics ; Eosinophils ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; genetics ; Gene Library ; Humans ; Male ; Middle Aged ; Nucleic Acid Hybridization

OBJECTIVETo explore the role of SHP-1 promoter methylation on the pathogenesis and progression in myelodysplastic syndromes (MDS) and its related mechanism.

METHODS63 MDS patients were divided into low-grade (LG) group and high-grade (HG) group according to IPSS score system. Bone marrow samples were collected. Methylation specific-PCR (MSP) were used to detect the status of SHP-1 promoter methylation in bone marrow (BM) samples from different risk MDS patients and MDS cell line, SKK-1. Western blot was used to detect signal transduction and activator of transcription (STAT3) activation in SKK-1 cell line and MDS patients.

RESULTSNo SHP-1 promoter methylation could be detected in healthy controls BM. Partially methylation was found in SKK-1 cell line. Methylation rate of SHP-1 gene promoter was found in BM of 24.2% of low-grade MDS patients and 63.3% of high-grade MDS patients, the difference between these two groups was statistically significant (P < 0.05); Patients were divided into different groups according to WHO subtype, chromosomal karyotype and blast cells in bone marrow, methylation rates of SHP-1 were significantly higher in RAEB-II, poor karyotype group and samples with 0.11-0.19 blast cells (P < 0.05); The phosphorylation protein of STAT3 was detected in SKK-1 cell line. The expression of phosphorylation STAT3 was significantly higher in HG group than in LG group (66.7% vs 18.2%) (P < 0.05). There was a significant correlation between SHP-1 promoter methylation and STAT3 phosphorylation.

CONCLUSIONAbnormal methylation of SHP-1 gene promoter might have tentative role in the pathogenesis and progression of MDS, which may be involved in STAT3 activation. Detection of SHP-1 promoter methylation may be helpful to evaluate the prognosis of MDS.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; Prognosis ; Protein Tyrosine Phosphatase, Non-Receptor Type 6 ; genetics ; metabolism ; STAT3 Transcription Factor ; metabolism ; Young Adult

BACKGROUNDRetinopathy of prematurity (ROP) has become one of the leading causes of visual loss in children. Vascular endothelial growth factor A (VEGF-A) is the principal stimulator of angiogenesis. VEGF was differentially spliced from exon 8 to exons 8a and 8b to form two families: the pro-angiogenic VEGFxxx family and the anti-angiogenic VEGFxxxb family. Previous research has shown variable effeteness of bevacizumab in inhibiting retinal neovascularization in ROP. This study aimed to investigate whether the effectiveness of this inhibition depends on the relative ratio of the two VEGF isoforms.

METHODSIntravitreal bevacizumab injection (IVB) was performed in the oxygen-induced-retinopathy (OIR) mice on postnatal day 12 (P12) (intravitreal phosphate buffered saline (PBS) injection as control). The Evans blue perfused retina were used to test the retinal neovascularization-leakage (NVL) area and non-perfusion (NP) area.

RESULTSThe retinal NVL and NP area in the IVB group were significantly smaller than the intravitreal PBS injection group (IVP group). On P17, the protein level of total VEGF isoforms was significantly inhibited compared to IVP group (P < 0.05) while VEGF(165)b isoform was slight reduced (P > 0.05). The switch from pro-angiogenic isoforms to anti-angiogenic isoforms after IVB could be found. The relative protein expression of VEGF(165)b isoform was significantly higher in IVB group than in IVP group (P < 0.05) on P17 which was correlated with the reduced ischemia-induced angiogenesis in OIR mice after IVB.

CONCLUSIONSThe anti-angiogenic effectiveness might depend on the relative high expression of VEGF(165)b after intravitreal bevacizumab injection. Anti-angiogenic therapy is a more effective therapy for ROP.

Angiogenesis Inhibitors ; administration & dosage ; Animals ; Animals, Newborn ; Antibodies, Monoclonal, Humanized ; administration & dosage ; Bevacizumab ; Disease Models, Animal ; Intravitreal Injections ; Mice ; Mice, Inbred C57BL ; Protein Isoforms ; analysis ; Retinal Neovascularization ; prevention & control ; Retinopathy of Prematurity ; drug therapy ; Vascular Endothelial Growth Factor A ; analysis

ObjectiveTo study the Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL)of the elderly and explore its clinical characteristics. MethodsThe clinical manifestation,laboratory examination and treatment of two cases of EBV positive DLBCL of elderly were lescribed.Results EBV positive DLBCL of elderly onset was often between 70-79 years,usually presented with lymphadenopathy and may be extranodal,may present with massive splenomegaly,secondary immune hemolytic anemia and often had B symptom.Pathologically,this disease was characterized by a proliferation of atypical large B cells with rich reactive cells,especially T cell.ConclusionImmunohistochemically,the tumor cells of EBV positive DLBCL of elderly present with CD20 or CD79a,EBV-encoded small RNA (EBER) positive.This disease has aggressive clinical course,a poor response to standard treatment.Rituximab may be effective in a short periods,but the long effect is limited.Overall survival is short.The cause of death is mainly respiratory failure due to factor infection.

Objective To investigate the effects of 3 different ventilation methods,including conventional mechanical ventilation (CMV),high frequency oscillatory ventilation(HFOV) and partial liquid ventilation (PLV),on the changes of inflammatory factors and pulmonary surfactant associated protein A (SP-A) in bronchoalveolar lavage fluid (BALF) in newborn piglets with acute lung injury(ALI).Methods Twenty-four newborn piglets,no more than 3 days old,were enrolled.After ALI made with saline lavage(38 ℃,35 mL/kg),newborn piglets were randomly assigned to 4 groups:control group (n =6,no ventilation),CMV group(n =6),HFOV group(n =6),and PLV group(n =6).Piglets were sacrificed after being ventilated for 24 h.Tumor necrosis factor-α (TNF-α),interleukin-8 (IL-8),interleukin-1 (IL-1) and SP-A in BALF were measured quantitatively by using enzyme-linked immunosorbent assay.Results In 3 groups using different ventilation methods,the population mean of TNF-o,IL-8,IL-1 and SP-A were statistically different (all P =0.000).SP-A in PLV group and HFOV group were higher than that in CMV group (all P ＜ 0.05),while IL-8,IL-1 and TNF-α in PLV group were lower than those in CMV group (all P ＜ 0.05),IL-8 and TNF-α in PLV group were lower than those in HFOV group (all P ＜ 0.05),IL-8 and TNF-α in HFOV group were lower than those in CMV group (all P ＜ 0.05).Conclusions Pulmonary inflammatory reaction was different in 3 ventilation groups.Compared with CMV and HFOV,PLV attenuated inflammatory reaction,so it could increase the expression of SP-A and decrease the degradation of SP-A.

Objective To investigate the pathogenic and clinical features of bloodstream infections in patients with hematological malignancies for improving clinical treatment.Methods A total of 92 patients with hematological malignancy and positive blood culture treated during the period from September 2011 to September 2016 were analyzed,including clinical manifestations,treatment and prognosis.The distribution and antibiotic resistance of pathogens were also investigated.Results Of the 92 patients with bloodstream infection,64.1％ had underlying agranulocytosis.All patients had fever.Septic shock was found in 45.7％ cases.Elevated procalcitonin was detected in 82.6％ cases.The 107 isolates from blood stream included 75 (70.1％) strains of gram negative bacteria,27 (25.2％) strains of gram positive bacteria,and 5 (4.7％) strains of fungi.Escherichia coli showed higher resistance rate to ceftriaxone (80.9％) and levofloxacin (91.3％).Klebsiella pneumoniae (31.2％) and Acinetobacter baumannii (50.0％) strains showed relatively high resistance to imipenem.Gram positive bacteria were still sensitive to vancomycin and linezolid.Overall,35 (38.0％) patients died.The initial empirical treatment regimen had significant impact on patient outcome (P＜0.05).The mortality rate of initial carbapenem-based empirical treatment was slightly lower (28.6％ vs 46.2％) than that of non-carbapenem-based initial regimen,but the difference was not significant (P=0.163).Conclusions The outcome is poor in patients with hematological malignancy complicated with bloodstream infection.The main pathogen is gram negative bacteria in such infections,associated with high antibiotic resistance.The emergence of carbapenem resistance is an issue of concern.The effectiveness of initial empirical therapy may have significant effect on patient outcome.

The aim of this study was to analyze chimerism, evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) by multiple short tandem repeat (STR) amplification using fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis. Peripheral blood and bone marrow in 27 patients who received myeloablative allogenetic cell transplantation were collected before and after transplantation in different times. 10 and 7 different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus/Cofiler plus PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype soft ware were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient. The results showed that donor chimerism was similar by the two methods. The median number of informative alleles was 6.3 (4 - 9) by Profiler Plus and 4.9 (2 - 6) by Cofiler Plus. The donor alleles appeared in 26 patients on day 28 post transplantation. One patient was not observed to appear donor alleles. 14 patients with 100% donor chimerism (DC) had stable engraftment and they still survive in free leukemia. 9 patients had unstable mixed chimerism (DC: 0% - 90.2%), and 5 of them relapsed after allo-HSCT, 6 patients died. Decrease of donor chimerism appeared prior to graft rejection and disease relapse. The incidence of GVHD was higher in group of full donor chimerism. It is concluded that dynamic monitoring donor chimerism by STR-PCR in combination with all auto-capillary electrophoresis is a valuable tool for predicting graft rejection, disease relapse and occurrence of GVHD, and provides a basis for early clinical intervention in the patients received allo-HSCT.
Adolescent ; Adult ; Female ; Graft vs Host Disease ; prevention & control ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Peripheral Blood Stem Cell Transplantation ; methods ; Polymerase Chain Reaction ; methods ; Recurrence ; Tandem Repeat Sequences ; Transplantation Chimera ; Transplantation, Homologous

OBJECTIVETo study the expression and clinical significance of P-gp, GST-pi and Topo II alpha in gastric and colorectal cancers.

METHODSThe expression of P-gp, GST-pi and Topo II alpha in 83 cases with gastric or colorectal cancer were examined by immunohistochemistry S-P.

RESULTSThe positive expression rates of P-gp, GST-pi, Topo II alpha in normal tissue and gastric and colorectal cancers were 69.9%, 65.1%, 50.6% and 83.1%, 85.5%, 45.8%, respectively. The positive rates of P-gp and GST-pi in gastric and colorectal cancer were significantly higher than those in normal gastric and colorectal tissue (P < 0.05). The expression of Topo II alpha in poorly differentiated cancers was significantly higher than that in well-and moderately differentiated cancers. There was no correlation between other items and clinicopathological parameters (P > 0.05).

CONCLUSIONP-gp, GST-pi and Topo II alpha play important role in multidrug resistance. Their mechanisms of drug resistance were different. The detection of expression of P-gp, GST-pi and Topo II alpha has an important guiding significance in chemotherapy for gastric and colorectal cancers.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; Adenocarcinoma ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Antigens, Neoplasm ; biosynthesis ; genetics ; Colorectal Neoplasms ; metabolism ; DNA Topoisomerases, Type II ; biosynthesis ; genetics ; DNA-Binding Proteins ; biosynthesis ; genetics ; Drug Resistance, Neoplasm ; Female ; Gastrointestinal Neoplasms ; metabolism ; Glutathione S-Transferase pi ; biosynthesis ; genetics ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Stomach Neoplasms ; metabolism

OBJECTIVETo investigate the oral health knowledge and habits of freshmen of Sichuan University and promote the prophylactic therapy of dental diseases in the university.

METHODSThe table and the standard on oral health survey authorized by WHO were employed in this study.

RESULTSOnly 13.1% of all the investigated freshmen had periodic oral health examination. 74.4% brush teeth twice per day, only 7.7% brush teeth over 3 minutes, 16.6% insist in fluoridated toothpaste, and only 6% had applied dental floss. 51.9% do not often take snack food and 94.7% are non-smokers; (4) Tests of prophylactic therapy of dental caries and periodontal diseases and oral health knowledge have poor results.

CONCLUSIONThe freshmen have limited knowledge of oral health and lack serious attitude toward oral cleaning behaviors.

Health Knowledge, Attitudes, Practice ; Humans ; Oral Health ; statistics & numerical data ; Oral Hygiene ; statistics & numerical data ; Students ; Universities