BACKGROUND:EpCAMhighCD44+ colon cancer stem cels have been isolated in recent years, and most of them are related to the study of isolating tumor stem cels in colorectal cancer cel lines. There are less reports on the function of colon cancer stem cels. OBJECTIVE:To isolate, screen and culture colon cancer stem cels from colorectal carcinoma tissues and to explore the protein expression and biological characteristics of colon cancer stem cels. METHODS: The primary cels were isolated from the colon cancer tissues. EpCAMhighCD44+ cels were detected and sorted by flow cytometry. Then, these cels were cultured in serum-free medium. After the cels were replanted subcutaneously into the mice, we observed the daily performance and tumor growth in the mice. When the tumor diameter was above 1 cm, tumor tissues were taken for immunohistochemical testing. The growth of EpCAMhighCD44+ cels cultured in the serum-free medium was observed; expressions of neutral epithelial mucin and CK-20 in EpCAMhighCD44+ cels were detected. RESULTS AND CONCLUSION: EpCAMhighCD44+ colon cancer stem cels were detected in the colon cancer tissues, and the proportion of EpCAMhighCD44+ cels in primary colon cancer cels was 1.7%-38%. After 24 hours of serum-free culture, there were a few of smal suspended cel spheres which were incompact. After 21 days, dense cel spheres with uniform size were found. The formation of transplanted tumor was found in the nude mice. Hematoxylin-eosin staining showed the transplanted tumor had the typical characteristics of colon cancer cels. The expression of neutral epithelial mucin and CK-20 was observed in al the transplanted tumors. Additionaly, the proliferation of EpCAMhighCD44+ cels could be promoted by 5-fluorouracil (10, 1, and 0.1 PPC) with time. These findings indicate that EpCAMhighCD44+ colon cancer stem cels in the colon cancer have a certain ability of proliferation, regeneration, differentiation, tumor formation and chemotherapy resistance.

Objective To clarify the role of cardiomyocyte apoptosis in cardiac function after transplantation of bone marrow cells. Methods A mycardial infarction model was induced in SD rats by left anterior descending artery ligation. 5?106 of bone marrow mononuclear cells were injected into peri-infarct zone (BMT group). In 4 or 8 weeks post-infarct, apoptosis of cardiomyocytes and expression of Bcl-2/Bax protein were determined by TUNEL or immunohistochemistry. Cardiac function was evaluated by echocardiography (UCG). Results TUNEL results indicated that apoptosis index of peri-infarct zone from the BMT group was reduced significantly when compared with the control group (4 wk: 0.094?6?0.017 vs 0.173?0.018, P

Objective To evaluate the effects of delayed transplantation of bone marrow mononuclear cells (BM-MNCs) after myocardial infarction (MI) on the expressions of bcl-2/bax mRNA and proteins in myocardial cells, and to explore the possible mechanism of cardiomyocyte apoptosis. Methods The MI rat model was reproduced by ligating the left anterior descending coronary artery. Two weeks later 5?106 of BM-MNCs were injected into the infarct zone and the peri-infarct zone (BMT group). TUNEL was used to determine the cardiomyocyte apoptosis, immunohistochemical method was employed to detect the expressions of bcl-2 and bax protein, and the technique of hybridization in situ was applied to assess the changes in of bcl-2/bax mRNA expression. Results TUNEL results indicated that apoptosis index of BMT group was lowered significantly compared with the control group (4 weeks: 0.095?0.017 vs 0.173?0.018; 8 weeks: 0.0916?0.014 vs 0.182?0.015, P

A strain of Prototheca species isolated from a case of meningitis was identified by routine morphologic and biochemical methods as well as amplification of the related genes, in which the 28S large-subunit (LSU) region of ribosomal RNA (rRNA) gene and intergenic space (ITS) were amplified with universal fungal primers. The small-subunit (SSU) rRNA gene was amplified with eukaryote-specific primers and Prototheca genus-specific primers. Then, compared the sequences with the ones posted on BLAST (www. ncbi. nlm. nih. gov/BLAST). The organism choice giving the closest match, up to 99%, was considered the most likely correct identification. It was found that this strain of fungus grew well at 25 ℃ or 37 ℃. Smooth,moist colonies with white color were observed on Sabouraud Dextrose Agar (SDA) and Potato Dextrose Agar (PDA). Microscopically, globular or ovoid cells, a number of round, ovoid shaped endospores could be observed. No hypha, ascus or blastic conidia was found upon cultivation on SDA. Based on the morphological characteristics, this isolate could be identified as Prototheca species. The identity with Prototheca wickerhamii was 2.9 % as demonstrated by the API 20C AUX system. Sequence analysis showed that the ITS gene was proved to be a complex structural region which was not suitable for the identification of Prototheca species, but the LSU and SSU rDNA regions showed 94% and 99.9% sequence identities with Prototheca zopfii var. hydrocarbonea (P. zopfii var. hydrocarbonea) respectively, indicating that the SSU rRNA gene sequence might be more reliable on than the LSU rRNA gene sequence for identification of Prototheca species.

Objective To assess the volume and function of right atrium(RA) in patients with chronic pulmonary hypertension(PH) by echocardiography and explore the influencing factors of these changes.Methods The study include 20 healthy participants as the control group and 57 patients with chronic PH as the case group.The patients were divided into three groups: low-grade PH, midrange PH and severe PH.Maximal RA volume index(VmaxI), minimal RA volume index(VminI), pre-atrial contraction RA volume index( VpreI), reservoir volume index(RVI), conduit volume index(CVI), contraction-chamber volume index (CCVI) were measured and calculated.The diameter of tricuspid valve and the late-diastole peak velocity of the bloodstream(Va) were measured,and then RA ejection force(RAEF) could be calculated.Systolic strain rate(RASRs), early diastolic strain rate (RASRe), and late diastolic strain rate (RASRa) of RA free wall were measured respectively.The correlation between pulmonary arterial systolic pressure (PASP), right ventricular ejection fraction(RVEF),the tissue Doppler of RV free wall(Ea/Aa) and the parameters of RA were analysed,and the main factors influencing the volume and function of RA were exploxed.Results With the rising of PASP, the RA volume changed obviously.And CCVI, RAEF, RVI, RASRs and the absolute value of RASRa increased gradually, but CVI decreased gradually.PASP, RVEF and Ea/Aa correlated with the parameters of RA.The main factors influencing the Vprel were EDVI, Ea/Aa and the course of disease,and VpreI and PASP mainly influenced the RAEF.Conclusions In patients with chronic PH the volume and function of RA changed,and some index of which was infuenced by PASP, RVEF, Ea/Aa and so on.

Siderophores were produced and secreted with marine microorganisms as the highly specific iron chelators. A method of liquid chromatography-tandem mass spectrometric detection coupled with solid phase extraction pretreatment was developed for the determination of siderophores in seawater. The samples were filtered through a 0. 22 μm membrane, extracted with an ENVI-18 cartridge and then eluted with methanol. The separation of the analytes was performed on a reversed phase SB-C18 column with a gradient elution program by using 0. 1% ( V/V) formic acid and methanol as the mobile phases. Qualitative analysis was performed in multiple-reaction monitoring mode. Good linearity (R2>0. 99) was obtained for Pyoverdines-Fe, Ferrichrome, Ferrioxamine E at the concentrations of 0. 001-3. 00 μg/mL, 0. 005-15. 00 μg/mL, 0. 001-3. 00 μg/mL, respectively. The instrumental detection limits and limits of quantification for the three analytes were 0. 08, 1. 76 and 1. 36 ng/mL; 0. 27, 5. 87 and 4. 53 ng/mL, respectively. The relative standard deviations were lower than 12%, while the recoveries were 12. 1%-18. 6% for Pyoverdines-Fe, 82. 0%-97. 7% for Ferrichrome, and 70. 0%-98. 3% for Ferrioxamine E.

A magnetic extraction sorbent based on Fe3 O4@poly ( methacrylic acid-co-ethylene dimethacrylate) ( Fe3 O4@MAED ) was synthesized using methacrylic acid and ethylene dimethacrylate as monomer and cross-linker, respectively. The magnetic sorbent was characterized by FTIR spectroscopy, elemental analysis and transmission electron microscopy. At the same time, the Fe3 O4@MAED was used to extract four benzoylurea pesticides in water and juice samples under magnetic dispersive solid phase microextraction ( MDSPME ) format. To obtain the optimal extraction conditions, several parameters, including the amount of sorbent, desorption solvent, extraction and desorption time, pH value, and ionic strength in sample matrix were investigated. Based on this, a fast, simple and sensitive method for the determination of benzoylurea pesticides in water and juice samples was developed by the combination of MDSPME with HPLC equipped with diode array detector. Under the optimized experimental conditions, the developed method possessed expected linear dynamic ranges, coefficients of correlation ( R2>0. 99 ) and sensitivity. The limits of detection (S/N=3) for target analytes were 0. 10-0. 19 μg/L in water and 0. 12-0. 30 μg/L in juice, respectively. The RSDs for intra-day were less than 7% and inter-day RSDs were less than 11%. The developed method was successfully applied to the determination of benzoylurea pesticides residues in water and juice samples and the recoveries of spiked target compounds were in the range of 69. 4%-118. 0%. The results demonstrated that the Fe3 O4@ MAED could extract benzoylurea pesticides effectively through multi-interactions including hydrophobic, hydrogen-bond and ion-exchange interactions between sorbent and analytes.

Objective:To investigate the possible mechanism of mesenchymal stem cells (MSC) secreting hepatocyte growth factor (HGF).Methods:① C57BL/6 mouse mesenchymal stem cells (mMSC) were cultured in vitro, and mMSC with high expression of chemokine receptor 7 (CXCR7) were transduced by lentivirus plasmid. Blank control group and empty carrier control group were set at the same time. After 20 generations of cell culture, the transfection efficiency was identified by fluorescence microscopy and flow cytometry. The mRNA expression levels of CXCR7 in mMSC were detected by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR). ② mMSC with passage number 4-6 were divided into MSC control group [MSC-blank group, 100 μg/L lipopolysaccharide (LPS) was added to wild-type MSC], highly expressed CXCR7 group (MSC-OE-CXCR7 group, 100 μg/L LPS was added to mMSC transduced by lentivirus plasmid with high expression of CXCR7), highly expressed CXCR7 control group (MSC-OENC-CXCR7 group, 100 μg/L LPS was added to mMSC transduced by no load lentivirus plasmid), CXCR4 inhibitor group (MSC-IE-CXCR4 group, 100 μg/L LPS was added to mMSC after 0.1 mg/L CXCR4 inhibitor TC14012 pretreatment for 24 hours), and CXCR4 inhibitor control group (MSC-IENC-CXCR4 group, 100 μg/L LPS was added to mMSC after DMEM culture medium with equal amount of TC14012 pretreatment for 24 hours). Cells in each group were collected after treatment with LPS, and mRNA expression of inhibitor of differentiation-1 (ID-1) was detected by RT-PCR. The cell supernatant was collected, and the levels of HGF were detected by enzyme linked immunosorbent assay (ELISA). Results:① The high expression of CXCR7 for mMSC which were transduced through lentivirus plasmid were successfully constructed detected by fluorescence microscope and flow cytometry. Compared with the blank control group, the expression of CXCR7 mRNA in the lentivirus with high expression of CXCR7 group was significantly increased (2 -ΔΔCt: 5.81±0.97 vs. 1.02±0.12, P < 0.05). There was no significant difference in CXCR7 mRNA expression between the empty carrier control group and the blank control group (2 -ΔΔCt: 0.95±0.22 vs. 1.02±0.12, P > 0.05). ②Compared with the MSC-blank group, high expression of CXCR7 in MSC-OE-CXCR7 group or inhibition of CXCR4 in MSC-IE-CXCR4 group could induce high expression of ID-1 mRNA in mMSC (2 -ΔΔCt: 5.56±0.66, 2.47±0.58 vs. 1.00±0.10, both P < 0.05) and increase HGF exocrine level (ng/L: 632.02±149.98, 217.21±40.53 vs. 108.53±24.62, both P < 0.05). However, there were no significant differences in ID-1 mRNA expression and HGF exocrine level of mMSC among MSC-OENC-CXCR7 group, MSC-IENC-CXCR4 group and MSC-blank group [ID-1 mRNA (2 -ΔΔCt): 1.01±0.27, 1.21±0.32 vs. 1.00±0.10, HGF (ng/L): 133.56±25.19, 107.11±25.30 vs. 108.53±24.62, both P > 0.05]. Conclusion:High expression of CXCR7 or inhibition of CXCR4 in MSC can increase the expression of ID-1 and promote the secretion of HGF, thus promoting pulmonary microvascular endothelial repair.

Objective To investigate the application value of the transesophageal echocardiography TEE in patients with nonvalvular atrial fibrillation about the size lobes morphology and function of left atrial appendage LAA Methods One hundred and forty-two patients underwent TEE were divided into nonvalvular atrial fibrillation group 98 cases and non atrial fibrillation group 44 cases The orifice diameter depth volume peak emptying velocity PEV of the LAA and the 1 eft atrial dimension LAD were measured The form and lobes of LAA thrombus and spontaneous echo contrast SEC in LAA were observed Results The LAA orifice diameter depth volume and LAD of patients with atrial fibrillation were significantly higher than those in the group without atrial fibrillation which showed statistical significance P < 0 05 Forty-one cases in atrial fibrillation group were found with the SEC and the number with thrombus in LAA was 22 The differences of PEV between chicken wings and non-chicken wings were statistically significant P <0 05 The SEC in LAA and the lobe number of LAA had no relevance Conclusions It was reliable to analyze the size morphologies lobes and hemodynamic parameters of LAA in patients with atrial fibrillation by TEE which provided reference for percutaneous LAA occlusion and anticoagulation therapy for the patients with atrial fibrillation.

Clustered regularly interspaced short palindromic repeats (CRISPR) is an acquired immune system existing in archaea and bacteria with the long-term process of evolutionary.CRISPR/Cas9 gene editing system is a new type of gene editing technology developed based on the system.CRISPR/Cas9 is a more efficient method for gene targeting than the previous methods.It has been successfully applied for gene-modified of eukaryotes since 2012,but the reports about pathogenic microorgaisms are rarely.Here,the research progress in the structure,mechanism of CRISPR/Cas9 system and its applications on pathogenic microorgaisms is reviewed.