Objective To investigate the difference of hypoxia-inducible factor 1a (HIF-1a) and vascular endothelial growth factor (VEGF) expression in endometrial cancer and para-carcinoma tissues, and to explore the clinical significance of hypoxia and the two proteins in the development and progression of endometrial cancer. Methods From Jan. 2011 to Dec. 2012, 128 patients with endometrial cancer underwent surgery in Tongji Hospital of Tongji University. The expression of HIF-1a and VEGF in cancer tissues and paired para-carcinoma tissues was detected using immunohistochemical method. The patients were followed up regularly, and the relationship between the expression of HIF-1a and VEGF and the prognosis of the patients was analyzed. The hypoxic cell model of human endometrial cancer was constructed to detect the expression of HIF-1a and VEGF proteins and observe the cell proliferation, invasion and apoptosis. Results The positive rates of HIF-1a and VEGF in cancer tissues were significantly higher than those in the para-carcinoma tissues (both P<0.05). The positive rate of HIF-1a was higher in the patients with lymph node metastasis, high histological grade, maximal tumor dimeter=4 cm or positive progesterone receptor (all P<0.05). The positive rate of VEGF was higher in the patients with lymph node metastasis, high histological grade, deep myometrial invasion, maximal tumor dimeter=4 cm, positive estrogen receptor, positive progesterone receptor or high pathological stage (all P<0.05). The 5-year overall survival rate of the patients with negative HIF-1a was significantly higher than that of the patients with positive HIF-1a (P<0.05), while there was no significant difference in the 5-year overall survival rate between the patients with negative and positive VEGF (P>0.05). In the hypoxic cell model of human endometrial cancer, the expression levels of HIF-1a and VEGF were significantly increased, cell proliferation and invasion were significantly increased, and the cell apoptosis was significantly reduced (all P<0.05). Conclusion HIF-1a and VEGF are related to the progress of endometrial cancer, and positive expression of HIF-1a indicates a poor prognosis.

A 2,4 -dichlorophenol degrading Pseudomonas strain GI241-1 was isolated from a soil sample. The dienelactone hydrolase gene, designated as dcpD which encodes dienelactone hydrolase involved in transforming cis-2-chloro-dienelactone into 2-chloromaleylacetic acid, was cloned from this bacterium strain. The gene cloning strategy was to construct genomic library after location of its neighbouring gene by Southem blot and to screen the aim transformant by dot blotting. Sequencing results showed that length of dcpD is 702bp. The sequence of dcpD and the deduced amino acid are different from the relative sequences registered in the GenBank.

2,4-Dichlorophenol is toxic and biorefratory organic pollutant. A 2,4-dichlorophenol degrading bacterial strain GT241-1, identified as Pseudomonas sp., was isolated from soil samples which was collected from drainage area of several 2,4-dichlorophenol producing factories. Strain GT241-1 had strong 2,4-dichlorophenol degrading ability, it could decompose 91% 2, 4-dichlorophenol of 90 mg/L within 48 hours at 25 - 30 degrees C, and could utilize 2,4-dichlorophenol, 2,4-dichlorophenoxyacetic acid (2,4-D), benzoate and catechol as sole carbon and energy source. Southern blot showed that 2,4-dichlorophenol hydroxylase gene (dcpA) of strain GT241-1 locates on the about 10kb EcoR I/Xba I fragment. This fragment was recovered, linked to the vecter pUC19 and transformed into the E. coli DH5alpha. A aim transformant, Z539, was obtained by dot blotting from about 1200 transformants. PCR and the sequencing results shew that the whole dcpA gene is contained within the 10kb EcoR I /Xba I fragment of pZ539. This fragment was shortened to about 2.4kb by HindmIII. The shorted fragment was subcloned to vecter pRSET-B to get a transformant BS1-12. The subcloned fragment was sequenced. Sequencing results showed that the whole length of the subcloned fragment containing dcpA is 2389bp and the nucleotide span of coding region is from number 276 to number 2072 (1797 bp), with ATG and TAA as start and stop codon respectively. The sequence analysis of dcpA and the deduced amino acid encoded by dcpA showed that they are different from the relative sequences registered in the GenBank. The subcloned fragment carry the promoter of dcpA, this can deduce from the fact that the upflow length of dcpA coding region is 275bp, and further confirmed by the 2,4-dichlorophenol hydroxylase activity measurement results. The 2,4-dichlorophenol hydroxylase activity of transformant Z539 and BS1-12 were detected, the results showed these transformants have 2,4-dichlorophenol hydroxylase activity. By comparison, the activity of these transformants were lower than that of the strain GT241-1.
Amino Acid Sequence ; Bacterial Proteins ; genetics ; metabolism ; Biodegradation, Environmental ; Chlorophenols ; metabolism ; Cloning, Molecular ; Environmental Pollutants ; metabolism ; Mixed Function Oxygenases ; genetics ; metabolism ; Molecular Sequence Data ; Pseudomonas ; enzymology ; genetics ; isolation & purification ; Soil Microbiology

The objective of study was to explore the influence of high mobility group box 1 (HMGB1) on migration of cord blood CD34(+) cells and their mechanism of migration. The expressions of receptor for advanced glycation end products (RAGE), toll-like receptor-2 (TLR2) and TLR4 were detected by flow cytometry. The CD34(+) cells in umbilical cord blood (CB) were enriched by MiniMACS and were exposed to various concentration of HMGB1 (10, 50, 100, 1, 000 ng/ml), then the migration effect of HMGB1 on umbilical cord blood (UCB) CD34(+) cell count was determined by microscopy, the chemotactic index was calculated. The CD34(+) cells untreated with HMGB1 were used as control. The results indicated that the purity of the isolated CD34(+) cells was more than 98%. The HMGB1 could promote the migration of CD34(+) cells, and the migration effect of HMGB1 on CD34(+) cells in certain concentrations gradually increased along with raise of concentration, the strongest effect was observed in concentration of 100 ng/ml, there was significant difference as compared with control (p < 0.01). Anti-RAGE antibody partially inhibited the migration effect of HMGB1 on CD34(+) cells. It is concluded that the HMGB1 in certain concentration can enhance migration of CD34(+) cells, which may be mediated through RAGE.
Antigens, CD34 ; Cell Movement ; drug effects ; Cells, Cultured ; Female ; Fetal Blood ; cytology ; drug effects ; HMGB1 Protein ; pharmacology ; Humans ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Signal Transduction

OBJECTIVETo evaluate the correlation of 64-multidetector-row CT (64MDCT) perfusion imaging with microvessel density(MVD) and vascular endothelial growth factor(VEGF) in colorectal carcinoma.

METHODS64MDCT perfusion imaging was performed in 33 patients with pathologically verified colorectal carcinoma. Time-density curves (TDC) were created from the region of interest (ROI) drawn over the tumor, target artery and vein by 64MDCT with perfusion functional software. The individual perfusion maps generated were for blood flow (BF), blood volume (BV), mean transit time (MTT) and permeability-surface area product (PS). MVD and VEGF expression of surgical specimens were examined by immunohistochemical staining with anti-CD34, anti-VEGF monoclonal antibody. MVD and VEGF were compared among the different types of TDC in colorectal carcinoma. The correlation of CT perfusion parameters with MVD and VEGF was also examined.

RESULTSTDC of colorectal carcinoma was divided into five types according to their shapes. MVD in the colorectal carcinoma was 22.61+/-9.01. VEGF staining was found in 25 of 29 tumors (86.2%). The score of VEGF expression was 4.15+/-1.09. No significant differences of MVD and VEGF expression among TDC types were found (F=2.59, 1.11, P>0.05). There were also no correlations of MVD and VEGF expression with any dynamic CT parameters (P>0.05).

CONCLUSION64MDCT perfusion imaging, MVD and VEGF may reflect angiogenic activity, but no significant correlations are found among them.

Adult ; Aged ; Colorectal Neoplasms ; blood supply ; diagnostic imaging ; Female ; Humans ; Male ; Microvessels ; Middle Aged ; Neovascularization, Pathologic ; Tomography, Spiral Computed ; methods ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

OBJECTIVETo explore the impact of IL-2- and IL-15-activated donor natural killer (NK) cell infusion on graft-versus-host-disease (GVHD) and graft-versus-leukemia (GVL) effect post allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThe C57BL/6 mice splenic NK cells were selected by microbeads, and then expanded in the media containing IL-2 and IL-15. The killing activity of NK cells was detected. In the leukemia mouse model, recipients (BALB/c) were intravenously inoculated with EL9611 leukemia cells 8 days before transplantation. Lethally irradiated BALB/c recipient mice were transplanted with 5 x 10(6) bone marrow cells (BMCs), or 5 x 10(6) BMCs plus 1 x 10(7) splenocytes with or without 1 x 10(7) activated NK cells. Additionally, NK cell infusion group mice were intraperitoneally injected with a mixture of IL-2 and IL-15 post transplant. Survival time, GVHD occurrence, lineage chimerism, TRBV spectra-typing were observed post transplant.

RESULTSThe purity of isolated splenic NK cells was 95.7% - 97.1%. The killing activity of NK cells after activation was increased by 3 times. GVHD did not occurred in allogeneic BMCs infusion group, whereas did from 1 week after transplant in allogeneic BMCs + splenocytes infusion group. The severity of GVHD in total body irradiation (TBI) experimental group was significantly lower than in splenocytes infusion group (P < 0.05). The survival time was 9.5 - 14.0 d in TBI alone conditioning group. In leukemia mouse model, 100 day survival rate was 10% the rest of them were died of leukemia while in experimental group, the more than 100 days survival rate was 80% (P < 0.01). PB NK cells at 2 week post-transplant were 4.8% in experimental group and 2.8% in control group. NK cells recovery in experimental group was earlier than that in control group (P < 0.05). TRBV reconstitution was faster in experimental group than in control group, moreover, the number of TRBV family expression was more in experimental group than in control group which mainly expressed monoclone or oligo-clone.

CONCLUSIONSDonor alloreactive NK cells can be efficiently expanded and activated with IL-2 and IL-15. Donor activated NK cell infusion and IL-2, IL-15 treatment can promote immune reconstitution, mitigate GVHD and reduce leukemia relapse.

Animals ; Cells, Cultured ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; Hematopoietic Stem Cell Transplantation ; Interleukin-15 ; immunology ; pharmacology ; Interleukin-2 ; immunology ; pharmacology ; Killer Cells, Natural ; cytology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL

The study was aimed to explore the influence of co-culture ex vivo of CD34+ cells from two units of cord blood (CB) on the homing-related adherent molecule expression of each other. Mesenchymal stem cells (MSCs) were obtained from human bone marrow. Two units of CB CD34+ cells were co-cultured on 12 Gy gamma-ray irradiated MSC layer. Their adherent molecule expressions were assessed by flow cytometry. The results showed that the purity of the isolated CD34+ cells was (98.25+/-0.93)%. After co-culture on MSC layer for 6 days, the proportion of CD34+ cells of each unit was dropped to (60.4+/-6.32)% and (60.2+/-5.12)% respectively, but there was no significant difference from the control groups. The expressions of CD44, CD62L, CD184 and CD26 on CD34+ cells of each unit remained unaffected. The expression of CD162 was downregulated and CD54 was first increased but then dropped to the level before co-culture. But there was no significant difference between the experimental and control groups. In conclusion, co-culture of CD34+ cells from two units of CB may have no effects on the adherent molecule expressions of each other.
Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; Cell Adhesion Molecules ; metabolism ; Coculture Techniques ; Fetal Blood ; cytology ; metabolism ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology

Objectve: To investigate the feasibility of establishing xenografted leukemia model by zebrafish, so as to provide the more direct model in vitro and experimental evidence for study of acute myeloid leukemia and screening of the drugs for targeting therapy.

METHODSAcute myeloid leukemia cell line KG-1a was labeled with red fluorescent dye-MitoRed, then the labeled cells were injected into the yolk sac of zebrafish embryos. Morphological observation, cell count and histopathological detection were used to analyse the infiltration and metastasis of KG-1a cells in zebrafish.

RESULTSKG1a cells could proliferate and gradually spread to the entire abdominal cavity of the zebrafish after KG-1a cells were injected into the yolk sac during 1-7, the results of cell counting in vitro also proved a significant proliferation of KG-1a cells in zebrafish, suggesting that the implanted leukemia stem cells could survive, proliferate and spread in zebrafish. Further study showed that the implanted cells could be transfered to the liver of zebrafish, these cells displayed the signature of KG-1a cells by hematoxylin-eosin(HE) staining.

CONCLUSIONSHuman acute myeloid leukemia cells KG1a can survive, proliferate and migrate in zebrafish, suggesting xenografted leukemia model of zebrafish has been successfully established. This model may be benefitcial for the study of acute myeloid leukemia and the screening of the drugs for targeting therapy of acute myeloid leukemia.

Objective To analyze the surveillance data on hemorrhagic fever with renal syndrome (HFRS) including the epidemiological characteristics and trend of the disease,in 2010.Methods Descriptive methods were conducted to analyze the surveillance data in 2010 which were collected from the internet-based National Notifiable Disease Reporting System and 40 HFRS sentinels in China.Results There were 9526 cases of HFRS reported in 2010 in the country with an annual morbidity of 0.71/105,which was higher than that reported in 2009.And the case fatality rate in 2010 was 1.24％.During the year 2010,most cases were reported in spring and autumn-winter season,with November as the peak month.The proportion of cases reported in autumn-winter season was higher than that in spring.The number of cases reported in males was higher than that in females among all the age groups,and similar pattern of mortality could be seen in most of the age groups.The percentage of cases over 60 years old had increased in recent years.Farmers were still under the highest risk.Density and the virus-carrying rate of animal hosts,as well as the infection rate were relatively stable and similar to the previous findings.As to the prevailing species,Apodemus agrarius and Rattus norvegicus were still the most common and leading animal hosts.However,the dominant species in sentinel of Yunnan were Rattus flavipectus and Eothenomys miletus respectively,and a new hantavirus called LUXV was found,namely Eothenomys miletus.Conclusion HFRS cases were widely distributed in most provinces of China,but cases mainly focus on certain areas and present the nature of aggregation.The risk of outbreak could not be ruled out for variety of factors.Population characteristics and seasonal fluctuation had been changing.

The objective of this study was to investigate clinical distribution characteristics and drug sensitivity of infection pathogens in hematology department wards of our hospital during recent two years. The isolation and cultivation of pathogens from samples were performed by routine methods. Drug sensitivity tests of bacteria were performed by Kirby-Bauer method. Drug sensitivity tests of fungi were performed by ATBFUNGUS Drug sensitivity strips. The results showed that 102 strains of pathogens were isolated from all detected samples. The composition ratio of Gram-positive bacteria, Gram-negative bacteria and fungi was 42.2%, 34.3%, 3.5%, respectively. 58.8% of pathogens were isolated from samples of malignant hematopathy patients. 27.5% were isolated from samples of the patients with fever of unknown origin (FUO). 51.0% of pathogens were isolated from samples of the patients who suffered from agranulocytosis or leucocytopenia. Isolated fungi were mostly sensitive to anti-fungal drugs. G+ bacteria were most sensitive to vancomycin. G- bacteria were most sensitive to imipenem. Most bacteria were resistant to multiple antibiotics. It is concluded that the infection in hematology department wards was related with many conditions, such as weakened resistance of patients, leucocytopenia or agranulocytosis, tumor loading, etc. The prompt microbiological examination and drug sensitivity tests are important to rationally select antibiotics, reduce infection incidence and mortality rate, and decrease the occurrence of drug resistant strains.
Cross Infection ; microbiology ; Fungi ; drug effects ; isolation & purification ; Gram-Negative Bacteria ; drug effects ; isolation & purification ; Gram-Positive Bacteria ; drug effects ; isolation & purification ; Hematologic Diseases ; microbiology ; Humans ; Imipenem ; pharmacology ; Microbial Sensitivity Tests ; Vancomycin ; pharmacology