Objective:To explore the influence of nephroblastoma over expressed gene (CCN3)on differentiation of bone marrow mesenchymal stem cells (BM-MSCs)into endothelial cells.Methods:BM-MSCs differentiation into endothelial cells induced by condition medium containing vascular endothelial growth factor (VEGF,50ng/ml)was regarded as positive control group.BM-MSCs differentiation into endothelial cells induced by condition medium con-taining recombined CCN3 protein (100 ng/ml)and VEGF (50 ng/ml)was regarded as CCN3 group,and BM-MSCs incubated in pure complete medium was regarded as negative control group.Immunofluorescence staining and semi-quantitative reverse transcription-polymerase chain reaction method (RT-PCR)were used to measure expression of von Willebrand factor (vWF)after 16d to evaluate endothelial cell differentiation,and semi-quantitative RT-PCR method was used to measured expression of Notch1 gene mRNA before and after induced BM-MSCs differentiation. Results:On 16d after induced BM-MSCs differentiation,some cells of positive vWF fluorescence staining were ob-served in positive control group,those of CCN3 group was significantly more than those of positive control group, and expression of vWF mRNA [ratio of optical density (OD): (0.550±0.090)]of CCN3group was significantly higher than that of positive control group (0.358±0.080),(P<0.01),and no vWF-positive cells were observed in negative control group;Besides,before induced differentiation,expression of Notch1 gene mRNA were low in three groups without significant difference between any two groups (P>0.05 all),on 16d after induced differentiation, expressions of Notch1 gene mRNA in positive control group (0.232±0.047)and CCN3 group (0.352±0.029)were significantly higher than that of negative control group (0.132±0.033),P<0.01 all,it of CCN3 group was signifi-cantly higher that of positive control group (P<0.01).Conclusion:CCN3 can enhance BM-BMCs differentiation into endothelial cells by activating Notch1 signal pathway.

Objective To investigate the role of detection of circulation alpha-fetoprotein(AFP) mRNA and telomerase activity in peripheral blood mononuclear cells(PBMC) of patients with hepatocellular carcinoma(HCC).Methods In patients with HCC,benign liver tumors,chronic liver diseases,and healthy volunteers,expression of circulation AFPmRNA and telomerase activity in PBMC were detected using nest reverse transcriptase-PCR(Nest RT-PCR) or PCR-ELISA respectively.Results(1) All the patients with benign liver tumors(n=10),chronic liver diseases(n=30),and healthy volunteers(n=30) had negative expression of circulation AFPmRNA,and the positive rate of circulation AFPmRNA in patients with HCC(73.3%,22/30)(all P

Objective To investigate the expression of oncogenes and tumor suppressor genes in hepatitis B virus (HBV) related hepatocellular carcinoma (HCC). Methods mRNA was extracted from cancerous and paracancerous tissues of 22 patients with HBV related HCC and synthesized into cDNA. The cDNA labeled with 33p dATP was hybridized for cDNA microarray each consisting of 3170 genes or expressed sequence tags (EST). The differential expression genes were searched in gene data base and verified using RT PCR. Results The differential expression of 1369 genes or EST was identified including 121 genes or EST altered 2 times or more in cancerous tissues compared with paracancerous tissues. Compared with paracancerous tissues, 88 showed overexpression and 33 genes were down regulated. The positive expression of transmembrane 4 superfamily member 1 (TM4SF1) in cancerous tissues was 86.4 % and could not be detected in paracancerous tissues (P

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BACKGROUND: Growth differentiation factor 5 (GDF-5) is an important factor to regulate the formation and development of the cartilage and bone, it plays a crucial role on the promotion of repairing bone, cartilage and tendon ligament injury. OBJECTIVE: To transfect eukaryotic expression plasmid pcDNA 3.1(+)/GDF-5 to bone marrow stroma stem cells of mouse and to check the expression of extracellular matrix and proteoglycan which relates with the cartilage formation and differentiation. DESIGN, TIME AND SETTING: An in vitro observation regarding cells was performed in the central laboratory of Wuhan Union Hospital between March and December in 2008.MATERIALS: Twenty Kunming specimen male mice were offered by Experimental Animal Center of Tongji Medical College of Huazhong University of Science and Technology. The eukaryotic expression plasmid pcDNA 3.1(+)/GDF-5 was preserved at the laboratory.METHODS: The man-ow stroma stem cells were isolated from mouse bone marrow and cultured in vitro with whole bone marrow adherence method. Passage 3 cells were incubated on 6-well plate and began to transfect when they were 90% confluent. Experiment was assigned into three groups: trensfection group underwent transient transfection of liposome-mediated pcDNA 3.1(+)/GDF-5 using LipofectamineTM2000; blank plasmid group was transfected with blank plasmid pcDNA 3.1(+); control group was added with equal volume of liposome and other protocols were the same as above. MAIN OUTCOME MEASURES: The transfection efficacy was identified success by the expression of GDF-5 gene and protein using RT-PCR and immunocytochemistry at 72 heurs following transfection, cartilage matrix Ⅱ collagen expression was determined as above methods. Then marrow stroma stem cells were cultured for additional two weeks to check expression of proteoglycan with alcian blue staining.RESULTS: In the transfection group, a 219-bp specific amplification band was visible, there were brown positive stain in the cytoplasm of marrow stroma stem calls; In blank plasmid group and control group, no GDF-5 trensfection, specific amplification band or obvious stain of cytoplasm was observed. In the transfection group, the collagen Ⅱ gene was detected to express at 225 bp, with brown yellow stain in cytoplasm; in the blank plasmid group and control group, no collagen Ⅱ gene expression or SP ～ stain was observed. Alcian blue staining results showed the transfected cells were stained blue while those in the blank plasmid group and control group were not metachromasia stained.CONCLUSION: Gene trensfection of pcDNA 3.1 (+)/GDF-5 to marrow stroma stem cells can significantly raise expression of collagen II and proteoglycan, and promote the chondrogenic differentiation of marrow stroma stem cells.

ObjectiveTo study the value of strain ratio from longitudinal section and cross section in differential diagnosis of benign and malignant thyroid nodules using ultrasonic elastography.MethodsA total of 118 patients with 153 thyroid nodules underwent examinations with real-time ultrasonic elastography to obtain the strain ratio from longitudinal section and cross section. Receiver operating characteristic (ROC) curves were used to evaluate the value of elastography. The nodules are divided into two groups, group one included nodules which were near the isthmus and group two included other nodules. Diagnostic results were compared with pathological results.ResultsThe area under the ROC curve (AUC) of the strain ratios for differentiating benign and malignant thyroid nodules were 0.906 and 0.844. There was no statistical difference (Z=1.542,P>0.05). The best diagnostic cut-off points of the strain ratios were 3.65 and 3.58. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value of the strain ratio from longitudinal section for differentiating benign and malignant thyroid nodules were 81.2%, 80.8%, 81.0%, 87.2% and 71.2% and those of strain ratio from cross section were 77.2%, 76.9%, 77.1%, 84.4% and 63.4%. In group one, the AUC were 0.903 and 0.830. There was no statistical difference (Z=1.125,P>0.05). The best diagnostic cut-off points were 3.30 and 3.28. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value of the strain ratio from longitudinal section for differentiating benign and malignant thyroid nodules were 86.2%, 68.8%, 80.0%, 83.3% and 73.3% and those of strain ratio from cross section were 78.6%, 58.8%, 71.1%, 75.9% and 62.5%. In group two, the AUC were 0.906 and 0.852. There was no statistical difference (Z=0.936,P>0.05). The best diagnostic cut-off points were 3.33 and 3.71. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value of the strain ratio from longitudinal section for differentiating benign and malignant thyroid nodules were 90.1%, 75.7%, 85.2%, 87.7% and 80.0% and those of strain ratio from cross section were 84.7%, 70.3%, 80.6%, 83.6% and 72.2%. ConclusionReal-time ultrasonic elastography is helpful to the differential diagnosis of thyroid nodules, but there is no statistical difference between the strain ratio from longitudinal section and cross section.

OBJECTIVE: To observe the effect of garlic oil combined with 5-FU induced apoptosis of adenoid cystic carcinoma cell line ACC-M. METHOD: Human salivary in adenoid cystic carcinoma cell line AC-M was cultured, divided into the experimental group (5-FU group, garlic oil group, garlic oil + 5-FU group) and the control group, to observe the growth activity of tumor cells by MTT methods; to analyse the changes of cell cycle and apoptosis rate by flow cytometry. RESULT: MTT experiments showed that 5-FU, garlic oil, garlic oil and 5-FU on ACC-M cells have inhibition in different concentration, with the increase of concentration and action time of the rise; Cell cycle analysis showed significant changes in flow cytometry. With the increase of concentration and the acting time, the G0/G1, phase of the cell ratio increased, S had no significant change, but G2/M phase cells decreased. Apoptosis rate display showed garlic oil combined with 5-FU induced apoptosis of ACC-M cells was significantly stronger than single group. CONCLUSION Garlic oil can effectively induce the apoptosis of adenoid cystic carcinoma cell line ACC-M. The effect of garlic oil combined with 5-FU on ACC-M cells was stronger than the garlic oil, 5-FU used alone.
Allyl Compounds ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Adenoid Cystic ; pathology ; Cell Line, Tumor ; Fluorouracil ; pharmacology ; Humans ; Sulfides ; pharmacology

OBJECTIVETo investigate the role of connexin 43-formed hemichannels in cell volume regulation induced by simulated ischemia/reperfusion (SI/R).

METHODSMouse cardiomyocytes isolated on a Langendorff apparatus with enzyme solution were aliquoted into control, SI/R and SI/R +octanol groups. Calcein-AM was used to stain the cells and the cell volume was measured with confocal microscope by stack scanning. Trypan blue was used to measure the cell viability after the treatments.

RESULTSCalcein-AM staining and cofocal microscopy yielded stable and reproducible results for cell volume measurement. Mouse cardiomyocytes subjected to simulated SI/R showed obvious cell swelling as compared with the control cells [(126∓6)% vs 100%, P<0.05], and octanol preconditioning significantly attenuated the cell swelling [(113∓6)%, P<0.05]. SI/R caused a significant reduction of the cell viability compared to the control cells [(19∓2)% vs (45∓3)%, P<0.01], and octanol preconditioning obviously reduced the viability of the cells with SI/R challenge [(31∓2)%, P<0.01].

CONCLUSIONConnexin 43-formed hemichannels are involved in the regulation of cardiomyocyte volumes induced by SI/R challenge, and octanol can alleviate the cell swelling to enhance the viability of the cardiomyocytes following SI/R.

Animals ; Cell Line ; Cell Size ; Cell Survival ; Connexin 43 ; metabolism ; Ischemic Preconditioning, Myocardial ; methods ; Mice ; Mice, Inbred C57BL ; Myocardial Reperfusion Injury ; metabolism ; pathology ; Myocytes, Cardiac ; drug effects ; pathology ; Octanols ; pharmacology

Objective To report our first clinical experience with a novel modified culotte technique for the treatment of true coronary bifurcation lesions. Methods The novel modified culotte technique (the mono-ring culotte) stenting was done in which the side branch (SB) stent was deployed firstly followed by ex vivo wiring of a most proximal cell of SB stent with the hard end of main branch (MB) wire. Secondly, the MB stent was deployed through the most proximal cell of SB stent. The procedure was ended with kissing balloon dilation. From June 2014 to March 2015, 15 patients with true coronary bifurcation lesion were treated with mono-ring culotte stenting in our center. Results The procedures were successful in all cases without procedural complication and in-hospital major adverse cardiovascular events. The procedural time was (34. 3 ± 9. 6) min, fluoroscopic time was (18. 1 ± 3. 8) min, and contrast volume was (112. 0 ± 24. 5) ml, respectively. Post-procedurally, the residual stenosis of the main and the side branch were (10. 0 ± 2. 5)% and (10. 2 ± 5. 3)% , respectively. Conclusions The mono-ring culotte stenting is safe and feasible for treatment of true coronary bifurcation lesions, and may be superior to the conventional culotte stenting.

Objective:To investigate the effect of surfactants on contact angle of pharmaceutical excipients and further study the effect of surfactants at different concentrations on the wettability of excipients and the effect of of surfactants and excipients at different proportions on the wettability of tablets. Methods:Different kinds of excipients were selected for tabletting,and surfactant solutions at different concentrations were prepared. The contact angle of the solutions on the surface of the tablets was measured by a contact angle tester. Results:The contact angle between the surfactants below the critical micelle concentrations and the tablets was the smallest and the wettability was the best. The change of the contact angle on the tablets with hydrophilic materials was less than that with hydropho-bic ones for most of the surfactants at the same concentration. Conclusion:After the addition of surfactants,the contact angle of excip-ient tablets significantly decreases, however, the contact angle will be stable after the amount of surfactants reaches a certain ratio, which shows significant reference value for tablet formula design.