Objective To investigate the value of detecting methylated SEPT9 gene (mSEPT9) in serum and stool in screening colorectal cancer in elderly people.Methods Subjects were divided into three groups, the colorectal cancer group (n=82), the colorectal polyps group (n=80) and the healthy control group (n=100).Real-time quantitative polymerase chain reaction (PCR) was used to detect mSEPT9 in peripheral blood and feces.The relationship between mSEPT9 and clinical and pathological characteristics of colorectal cancer was analyzed.The agreement between serum and stool results was analyzed.Results The positive rate of serum mSEPT9 was 73.2％ in the colorectal cancer group, 6.3％ in the colorectal polyps group and 4.0％ in the healthy control group, with statistical significance between the colorectal cancer group and the other two groups (P＜0.01).The sensitivity and specificity for plasma mSEPT9 detection in screening colorectal cancer were 73.2 ％ and 95.6％, respectively.The positive rate was higher in patients with tumor maximum diameter over 3.0 cm, invasion of the entire serosa layer, lower differentiation or advanced clinical staging (P＜0.05).The detection results of plasma mSEPT9 were in complete agreement with those of stool mSEPT9 from the same patients.Conclusions The expression of mSEPT9 is high in elderly patients with colorectal cancer and has a high sensitivity and specificity for PCR detection.Complete agreement can be seen in results from plasma and feces.Detecting mSEPT9 can be a valuable approach to screening colorectal cancer in the elderly.

Objective:To compare the two-dimensional electrophoresis(2-DE) maps of rat spinal cord protein extracted by two different solution systems.Methods: Adult rat spinal cord protein was precipitated with 10% trichloracetic acid in acetone and resuspended in 8 mol/L urea plus 4%CHAPS (A solution) or, 5 mol/L urea, 2 mol/L thiourea, 2%CHAPS plus 2%SB3-10 (B solution). One hundred and fifty micrograms of protein was loaded on 18 cm IPG strip holder and run isoelectric focusing electrophoresis as the first dimension, then horizontal SDS-PAGE as the second dimension. Protein spots were visualized by silver stain.Results:There were 1 059 and 1 023 protein spots in each map, of which 790 spots were matched in two maps. There were 269 and 233 spots exclusively extracted by A and B solutions, respectively. Taken together, 1292 different spots were totally obtained by A and B solutions.Conclusion: Integrating protein spots extracted by different solution systems is beneficial for achieving intact 2-DE map of tissues.

Objective: To explore the expression of SLAMF6 on CD8(+) T cells in patients with severe aplastic anemia (SAA) and its correlation with disease immune status. Methods: By flow cytometry (FCM), SLAMF6 expression level in peripheral blood CD8(+) T cells was detected in 21 patients with SAA and 15 normal controls respectively from February 2017 to April 2018. The correlation between SLAMF6 expression level and hematopoietic functions, including HGB, PLT, the neutrophil granulocyte and reticulocyte absolute value in peripheral blood, hyperplasia degree (percentage of granulocytes, erythrocytes, lymphocytes and megakaryocytes in bone marrow) and perforin, granzyme B, IFN-γ expression level in CD8(+) T cells were evaluated. To further confirm the effect of SLAMF6 on CD8(+) T cells, anti-SLAMF6 Ab was used to block SLAMF6 pathway (IgG as control), and FCM was used to detect the perforin, granzyme B, and IFN-γ production of CD8(+) T cells. Results: The expression of SLAMF6 on CD8(+) T cells in untreated SAA patients[(56.29±12.97)%]was significantly lower than that of normal controls[(80.96±7.36)%](t=-7.672, P<0.001). The expression of SLAMF6 on CD8(+) T cells in SAA patients were positively correlated with the HGB, PLT, the neutrophil granulocyte and reticulocyte absolute value in peripheral blood, percentage of granulocytes, erythrocytes in bone marrow (all P<0.05), but they were negatively correlated with the percentage of lymphocytes in bone marrow, and the expression of perforin, granzyme B, and IFN-γ of CD8(+) T cells (all P<0.05). After blocking SLAMF6 pathway by anti-SLAMF6 Ab, the expression levels of perforin, granzyme B and IFN-γ in SAA patients were significantly higher than those in the untreated group, and the differences were statistically significant (all P<0.05). Conclusions: SLAMF6 is significantly down-regulated on CD8(+) T cells in SAA patients, which may act as a negative immunoregulatory molecule participating in the mechanism of SAA by affecting the functional molecules secretion on CD8(+) T cells.
Anemia, Aplastic ; Bone Marrow ; CD8-Positive T-Lymphocytes ; Flow Cytometry ; Humans ; Perforin ; Signaling Lymphocytic Activation Molecule Family

OBJECTIVETo investigate the viral pathogens of acute respiratory infection (ARI) in hospitalized children from Suzhou of China.

METHODSThe nasopharyngeal aspirate samples were obtained from 1,668 hospitalized children with ARI. Common respiratory viruses, including respiratory syncytial virus (RSV), influenza virus A and B, parainfluenza viruses 1, 2 and 3 and adenovirus, were detected using direct immunofluorescence. Human metapneumovirus (hMPV) gene fragments were detected by RT-PCR.

RESULTSViral agents were identified in 597 cases (35.8%). RSV was the most frequent (17.6%). RSV infection is more common in children less than 1 year old. A highest detection rate of RSV was found during winter and spring. hMPV was detected in 10.6% of the cases, with a peak detection rate between March and May. Single viral infection was found in 561 cases (33.6%) and mixed viral infection in 36 cases (including 27 cases at age of less than 1 year). RSV and hMPV co-infection was common (n=22).

CONCLUSIONSRSV is common pathogen of ARI in children from Suzhou. RSV viral activity peaks during winter and spring. The children at age of less than 1 year are susceptible to RSV. hMPV is also an important pathogen of ARI, with a peak detection rate between March and May. Mixed viral infection is common in children less than 1 year old.

Acute Disease ; Age Factors ; Child ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Metapneumovirus ; isolation & purification ; Nasopharynx ; virology ; Respiratory Syncytial Virus, Human ; isolation & purification ; Respiratory Tract Infections ; epidemiology ; virology ; Seasons

OBJECTIVETo evaluate the clinical and laboratory features of pediatric inv(16) acute myeloid leukemia (AML) retrospectively.

METHODDual color fluorescence in situ hybridization (D-FISH) using a LSI CBFβ inv(16) break apart probe labeled by Spectrum red and Spectrum green was performed in 15 acute myeloid leukemia cases, including 13 cases with or without abnormal eosinophils but with positive core binding factor β (CBFβ)-MYH11 fusion transcript detected by RT-PCR, and 2 cases with trisomy 8 (+8). The results were compared with the morphology, immunophenotype, karyotype and RT-PCR.

RESULTMorphologically, 12 cases were diagnosed as M(4)EO, 2 as M(4), and 1 as M(2a). Immunophenotypically, all 13 AML cases with inv(16) showed positive expression of CD(13) and CD(33), but without the expression of any lymphoid lineage antigens. Karyotyping analysis with G-banding detected inv(16) in 10 AML cases, including 9 M(4)EO cases and 1 M(2a), but only 5 positive cases were detected using R-banding technique. Among them, 2 cases had simultaneous +8 and trisomy22 (+22), one had +22 only in addition to inv(16). D-FISH revealed a CBFβ-MYH11 rearrangement in 13 cases of AML with positive RT-PCR results, and the mean positive rate of cell detection was 55.15% (range 37.0% - 86.0%). The complete remission rate (CR) and median survival period in this series of inv(16) AML were 81.5%and 11 months, respectively, of whom, 8 cases were still in CR. Relapse and karyotypic evolution were seen in case 5 with +8, +22 in addition to inv(16).

CONCLUSIONAML with inv(16) is a special subtype. Most cases belong to M(4)EO. Its prognosis is good in general, but it seems to be an unfavorable feature for AML with inv(16) and +8, +22 simultaneously, especially with karyotypic evolution. For detection of inv(16), G-banding technique is evidently superior to R-banding technique. D-FISH combined with RT-PCR are more sensitive and reliable than chromosome banding analysis.

Adolescent ; Child ; Child, Preschool ; Chromosome Deletion ; Chromosome Inversion ; Chromosomes, Human, Pair 16 ; genetics ; Eosinophilia ; pathology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Male ; Prognosis ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction

To study the effects and possible mechanism of Vitamin K(2) (VK(2)) in the treatment of MDS-JSN04 cells, the changes of morphologic features of MDS-JSN04 cells were investigated by cytomorphology, the apoptosis of MDS-JSN04 cells was observed by transmission electron microscope; cellular proliferation was determined by the MTT assay; cell apoptosis, cell cycle shift and expression of myeloid-specific differentiation antigen (CD11b, CD13) were analyzed by flow cytometry (FCM). The expression of apoptosis-related genes bcl-2, survivin and bax were detected by retrotranscriptase polymerase chain reaction (RT-PCR); the activity of caspase-3 was determined by chemiluminescence assay. The results showed that the typical apoptotic morphological features appeared in cells treated with VK(2) for 72 hours; VK(2) induced apoptosis of MDS-JSN04 cells and in a dose-and-time-dependent manner, G(0)/G(1) cell arrest and significantly down-regulated the expression of bcl-2 and survivin, but had no effect on the expression of bax; the activity of caspase-3 significantly increased. It is concluded that VK(2) induces apoptosis of MDS-JSN04 cells through activating caspase-3 pathways and the apoptosis-related genes bcl-2, survivin may play an important role in this process.
Apoptosis ; drug effects ; CD11b Antigen ; analysis ; CD13 Antigens ; analysis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; Luminescent Measurements ; methods ; Microscopy, Electron, Transmission ; Microtubule-Associated Proteins ; genetics ; Myelodysplastic Syndromes ; genetics ; metabolism ; pathology ; Neoplasm Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Vitamin K 2 ; pharmacology ; bcl-2-Associated X Protein ; genetics

Objective To evaluate the role of cerebral state index(CSI),burst suppression (BS)and electromyograph(EMG)in monitoring coma/consciousness depth and damage degree of brain. Methods CSM was done in 50 cases with brain injury and coma to analyze its relation with physical reflection,auditory evoked potential(AEP),Glasgow coma score(GCS)and Glasgow outcome scale (GOS).Results As scale range meaning from consciousness to deep coma and to brain death,CSI 0- 100 was positively correlated with coma depth,coma score of GCS and physical reflection.CSI changes under invariable ache stimulation in combination with BS and EMG can accurately estimate prognosis and quantify changes of brain function.Conclusions The quantifiable digit of coma/consciousness depth and damage degree in brain function by CSM can attain real time judgment of dynamic evolvement course of coma and objective guide clinical therapy and assure prognosis,as will change absolutely scoring coma/ consciousness depth and prognosis under current state of artificial diversity and lacking objective evi- dences.

Objective To explore the effect of ox-LDL,MCP-1,CD68 on the survival of arteriovenous fistula in radial arteries of uremic patients.Methods Segments of radial arteries were obtained from 23 uremic subjects (29～68 years old) in the initial operation of arteriovenous fistula prior to hemodialysis.The deposit of ox-LDL and the expression of MCP-1,CD68 on the vascular wall were measured by immunohistochemistry.The survival time of arteriovenous fistula was followed by survival analysis.Results COX regression revealed that each of these risk factors,ox-LDL,MCP-1, CD68,played an important role in the survival time of arteriovenous fistula when they entered the model independently.The hazard ratios were 1.008 (P=0.008,95% CI:1.002064～1.014104), 1.007(P=0.O00,95%CI:1.003853～1.010966),and 1.098496 (P=0.000,95%CI:1.047909～1.151526)respectively.When all the three factors entered the COX regression model,the whole model was still founded.MCP-1 and CD68 still played important roles in the survival of arteriovenous fistula.The hazard ratios were 1.006(P=0.025) and 1.113(P=0.001) respectively.With the hazard ratio of 0.997,ox-LDL did not reach the statistic significance (P=0.414).Conclusions The more deposit of ox-LDL and the more expression of MCP-1,CD68 on the vascular wall,the more shortened survival time of arteriovenous fistula.Particularly,the inflammation is the independent risk factor for the prognosis of arteriovenous fistula in uremic patients.

This study was aimed to establish a cytokine-independent human myelodysplastic cells line from bone marrow of a patient with MDS-CMML. This cell line was incubated in mixed culture of RPMI 1640 and DMEM with 15% bovine serum, but without cytokines; its biological characteristics were identified by morphology, surface marker profiles, cell proliferation, differentiation and apoptosis. The results showed that the established cell line could not depend on cytokines for long-term survival and growth, and could differentiate into colony-forming unit-macrophage, colony-forming unit-megakaryocyte. In conclusion, a cytokine-independent human myelodysplastic syndrome cell line, named MDS-JSN04 (MDS Nanjing Jiansu 04), was established. Its partial biological characteristics were identified and clarified.
Antigens, CD19 ; analysis ; Antigens, CD20 ; analysis ; Apoptosis ; drug effects ; Bone Marrow Cells ; metabolism ; pathology ; ultrastructure ; CD79 Antigens ; analysis ; Cell Differentiation ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Culture Media ; pharmacology ; Cytokines ; pharmacology ; Flow Cytometry ; HLA-DR Antigens ; analysis ; Humans ; Lewis X Antigen ; analysis ; Male ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; Sialic Acid Binding Ig-like Lectin 2 ; analysis ; Time Factors