Objective To study the levels of IL 6 production in peripheral blood mononuclear cells (PBMC) in 25 patients with inactive and active (include mild、 moderate and severe) ulcerative colitis(UC) and age, sex matched 20 healthy subjects.Methods Peripheral blood monocytes (PBMC) were stimulated with PHA for 48 hours to induce IL 6 production. IL 6 content in the culture medium was assayed by using B9 cells. Results IL 6 production was significantly increased in PBMC from active UC. There was an increasing tendency with severity of disease. But no significant difference was found between the extent of the lesious. Conclusions The levels of IL 6 production in PBMC can be an indicator of the activity of UC.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are in advantages of easy collection and amplification in vitro, but the culture and induction in vitro still need to be modified. OBJECTIVE: To investigate the differentiated potency of BMSCs into ostcoblasts by modified in vitro culture methods and inductor matching. DESIGN: Observational study. SETTING: Department of Medical Laboratory, General Hospital of Guangzhou Military Area Command of Chinese PLA. MATERIALS: This study was performed in the Stem Cell Tissue Engineering Laboratory, Department of Medical Laboratory, Genera Hospital of Guangzhou Military Area Command of Chinese PLA from July 2004 to June 2006. Twenty adult SD rats of clean grade, irrespective of gender, weighing 140-180 g were provided by Animal Experimental Center, General Hospital of Guangzhou Military Area Command of Chinese PLA. The experimental animals were disposed according to ethical criteria. Β-sodium glycerophosphate, dexamethasone, and vitamin C were provided by Sigma Company, USA; goat-anti-rat osteocalcin antibody by DSL Company, USA; S-P immunohistochemical kit by Maixin Biotechnology Developing Co., Ltd., Fujian. METHODS: Cells were cultured and induced into osteoblasts by modified methods. Separation and culture of BMSCs: By anesthesia, bone marrow of bilateral femur and tibia was separated to prepare single cell suspension; subsequently, the suspension was inoculated in culture bottle, and the culture media was semi-quantitatively changed after 48 and 96 hours in order to get rid of non-adherent hematopoietic cells. The liquid was changed every three days to further get rid of non-adherent cells. At 80% cell confluence, the medium was digested with 0.25% trypsin and cells were subcultured in the ratio of 1:2. MSCs in the second generation were inoculated in 6-well culture plate and glass fiat plate; after 48 hours, basic culture fluid was removed. Differentiation of BMSCs into osteoblasts: Subcultured BMSCs differentiated into osteoblasts induced by dexamethasone (10 mmol/L), β-sodium glycerophosphate (10-7 mol/L), and vitamin C (50 mg/L). MAIN OUTCOME MEASURES: Ten days after differentiation by modified culture methods and inductor matching, alkaline phosphatase activity was measured with Ca-Co technique. Twelve days after culture, osteocalcin secretion was detected with immunohistochemical method. Two weeks after culture, cell mineralization was detected with Von kossa staining. RESULTS: Alkaline phosphatase activity: Alkaline phosphatase staining of cells was apparent; gray-black particles or massive precipitations were observed in cytoplasm after positive reaction; regions expressing alkaline phosphatase activity were brown-black. Osteocalcin secretion: Osteocalcin was apparently positive; nucleus was blue; cytoplasm was brown. Cell mineralization: Induced cells grew layer by layer, and adiaphanous nodus was observed at the same time. Black mineralized nodus granules in various sizes were observed after Von kossa staining, and this suggested that mineralized apposition occurred. CONCLUSION: BMSCs may be successfully cultured and induced into osteoblasts by modified culture method.

BACKGROUND:With the potential of multi-directional differentiation and high proliferation, bone marrow mesenchymal stem cells (BMSCs) have wide application prospect in tissue engineering. OBJECTIVE:To investigate changes in cells and bioactivity in the differentiation from BMSCs into osteoblasts. DESIGN, TIME AND SETTING:A randomized control experiment was performed at the Laboratory of Stem Cells and Tissue Engineering, Department of Medical Experiment, General Hospital of Guangzhou Military Area Command of Chinese PLA from July 2004 to June 2006. MATERIALS:Twenty adult SD rats of both genders were used for bone marrow collection. METHODS:Rats were equally and randomly divided into an experimental group and a control group. BMSCs were isolated from adult rats by modified bone marrow culture method. BMSCs were inoculated in basic medium containing 10% fetal bovine serum, high glucose DMEM, 100 U/L penicillin, 100 U/L streptomycin and 2 mmol/L glutamine in the control group. BMSCs were inoculated in conditioned medium (basic medium, 10 mmol/L β-glycerophosphoric sodium, 10-7 mol/L dexamethasone, 50 mg/L vitamin C). Cell slide was prepared. MAIN OUTCOME MEASURES:Inverted microscope and transmission electron microscope were used to observe cell appearance. Gomori modified calcium-cobalt method was applied to do alkaline phosphatase staining. Immunocytochemistry was employed to measure osteocalcin expression. RESULTS:Forty-eight hours after inoculation, a mass of BMSCs adhered to a wall. Seventy-two hours later, BMSCs proliferated and well grew. Seven to nine days later, cells were grown to 80% confluency. Transmission electron microscope showed BMSCs with big cell nucleus and immature appearance. After in vitro osteoblast induction, BMSCs proliferated, aggregated, had node and mineralized. One week later, alkaline phosphatase activities were expressed in BMSCs. Two weeks later, osteocalcin expression was detected. CONCLUSION:After one week of in vitro osteogenic induction, BMSCs enter the process of osteoblast transformation, remain proliferative activities, and can be used as seed cells in bone tissue engineering

Adult ; Female ; Humans ; Intestinal Neoplasms ; Male ; Middle Aged ; Sarcoma, Myeloid

Objective To estimate reliability of magnetic resonance elastography (MRE) in measuring liver stiffness of volunteers and patients with chronic liver disease and to assess inter-rater consistency.Methods MRE was performed on a 3.0 T scanner in all subjects,including 24 volunteers (control group) and 64 patients with liver disease (chronic liver disease group).Liver stiffness was measured blindly by two raters.The pathological fibrosis score was applied as a standard reference for liver fibrosis in 22 patients.The intraclass correlation coefficient (ICC) was used to evaluate inter-rater reliability.The differences of liver stiffness between two groups were evaluated using non-parametric MannWhitney U test.Spearman analysis was used to analyze the correlation between fibrosis stages and liver stiffness.Results The intraclass correlation coefficient of liver stiffness was perfect (ICC =0.99,P ＜ 0.01)between two raters.There was significant difference of mean stiffness between control group and patient group (U =90.5,P ＜0.01) with(2.35 ±0.34) kPa and(4.17 ± 0.47) kPa,respectively.The correlation between fibrosis stage (3,3,5,5 and 6 patients in fibrosis stage S0,S1,S2,S3 and S4) and stiffness (2.13,3.25,3.82,5.45 and 7.35 kPa) was very strong (r =0.96,P ＜0.01).Conclusion MRE is a reliable and promising tool to measure liver stiffness and to assess liver fibrosis.

Objective To investigate the regulation effect of promoter methylation of deathassociated protein kinase (DAPK) on mRNA and protein expression of DAPK in tissue of primary gastric cancer (GC). Methods The cancerous and noncancerous samples from 62 patients with GC were determined by RT-PCR for mRNA expression of DAPK. The DAPK promoter methylation was detected by methylation-specific PCR. The protein expression of DAPK in 34 patients with methylation was determined by Western blot. Results mRNA and protein expre.ssions of DAPK in cancerous tissues were reduced significantly compared to noncancerous tissues (0. 2863d±0. 2027 vs 0. 57364±0. 1968,0. 2616±0. 0913 vs 0. 65294±0. 1808, P＜0.01). Methylation frequency of DAPK in cancerous tissues was higher than that in noncancerous tissues (54.8% vs 17.7%, P＜0.01). Furthermore, DAPK mRNA expression was decreased in methylation group compared to unmethylation group (0.1399±0. 0835 vs 0. 46404±0. 1569, P＜0. 01). Moreover, a significant correlation was demonstrated between the TNM stage and DAPK promoter methylation (P = 0. 04). Conclusion Expression of DAPK is down-regulated in cancerous tissues at mRNA and protein levels. Low expression of DAPK is associated with hypermethylation of the promoter of DAPK gene.

Adult ; Agriculture ; Antibody Formation ; Female ; Humans ; Immunity, Cellular ; Male ; Middle Aged ; Occupational Exposure

Objective To investigate an association between Toll-like receptor(TLR)4 Asp299Gly or CD14 C-260T polymorphisms and colorectal cancer in Chinese patients.Methods By a method of polymerase chain reaction-based restriction fragment length polymorphism(PCR-RFLP), TLR4 Asp299Gly and CD14 C-260T polymorphisms in unrelated 110 patients with colorectal cancer and 160 healthy controls in Chinese Han race population were genotyped.Results Significant differences of CD14 genotypes were found between healthy controls and the patients with colorectal cancer.The fre- quency of the CC genotype in healthy controls was extremely lower than that in the group of colorectal cancer(15.6% vs.31.8%,P=0.0027,OR=0.3968,95%CI:0.2209-0.7129)while the frequency of the CT genotype in healthy controls was significantly higher than that in the group of colorectal cancer (48.1% vs.30.9%,P=0.0056,OR=2.074,95%CI:1.246-3.452).No TLR4 Asp299Gly mutant genotype was detected in any patients and healthy controls in Chinese Han population.Conclusion The polymorphism of CD14 C-260T but not TLR4 Asp299Gly is associated with colorectal cancer in Chinese patients.

Objective:To detect the inhibitory effects of CAL-101, a selective inhibitor of phosphoinostitide-3'-kinase delta (PI3Kδ), on Burkitt's lymphoma cell line Raji and diffused large B-cell lymphoma cell line SUDHL-10 and elucidate its relative mechanism. Methods:Raji and SUDHL-10 cells were treated with various concentrations of CAL-101. Methyl thiazolyl tetrazolium (MTT) assay was performed to determine the inhibitory effect of CAL-101 on lymphoma cells, and cell apoptosis was measured by Annexin V/PI and DAPI staining. Migration assays were performed with transwell to detect the migration of lymphoma cells derived from the stromal cell line HK. Western blot was used to detect the phosphorylation status of the ERK pathway. MTT and CalcuSyn software analyses were preformed to detect whether or not combining CAL-101 with bortezomib induces synergistic cytoxicity. Results:CAL-101 at con-centrations of 5, 10, 15, and 20μmol/L inhibited cell proliferation in a dose-dependent manner. The proliferation rates of the Raji cells treated with 5, 10, 15, and 20μmol/L for 48 h were 29.17%± 1.23%, 38.15%± 1.51%, 46.46%± 1.78%, and 55.8%± 2.01%, respec-tively, which were significantly higher (P<0.05) than that of the control group (1.15% ± 0.02%). Similar results were found in the SUDHL-10 cells after treatment with CAL-101 (P<0.05). CAL-101 also exerted an apoptotic effect on the lymphoma cells. The apop-totic rates of the Raji cells treated with CAL-101 for 21 h were 22.69%± 3.83%and 49.96%± 7.36%, respectively, which were signifi-cantly higher (P<0.05) than that of the control group (5.23%± 2.04%). Similar results were found in the SUDHL-10 cells (P<0.05). Treatment with 5 and 10 μmol/L CAL-101 dose-dependently inhibited the migration activity of lymphoma cells to stromal cells (P<0.05). Western blot analysis showed that the expression level of ERK phosphorylation protein was significantly downregulated in the cells treated with CAL-101. A synergistic effect between CAL-101 and bortezomib was verified. That is, these two drugs can signifi-cantly inhibit the proliferation of lymphoma cells with CI values less than 1. Conclusion:The PI3Kδ-specific inhibitor CAL-101 sup-pressed the proliferation of Raji and SUDHL-10 cells, induced apoptosis, and inhibited stromal cell-derived migration. This inhibitory effect may be induced by blocking the ERK pathway. Overall, our study indicated that CAL-101 is a novel and potential agent in the therapeutic strategy against aggressive B-cell lymphoma.

BACKGROUND:Flexible intramedulary nailing and external fixation for pediatric femoral shaft fractures are two common methods in the clinic. It remains controversial which fixation methods are better. OBJECTIVE:To systematicaly evaluate the therapeutic effects of flexible intramedulary nailing and external fixation for pediatric femoral shaft fractures. METHODS: A computer-based search was performed on PubMed, Embase, Medline, and Cochrane library for literatures on clinical controled trials of flexible intramedulary nailing and external fixation for pediatric femoral shaft fractures published before November 25, 2014. Literature language was not limited. The age of subjects was from 3 to 15 years. Modified Jadad was utilized to assess methodological quality of the included studies. Meta-analysis was carried out using Stata 12.0 software. RESULTS AND CONCLUSION:Six papers involving 237 patients were included. Meta-analysis results showed that compared with external fixation, a low incidence of overal complications [relative risk (RR)=0.30, 95% confidence interval (CI): 0.19-0.46;P < 0.001] and pin-hole infection (RR=0.286, 95%CI: 0.13-0.61;P= 0.001), but a high risk of needle tail irritation (RR=1.86, 95%CI: 1.35-2.56;P < 0.001) were found folowing flexible intramedulary nailing. No significant differences in other complications were found between the two groups. These results confirm that compared with external fixation, elastic intramedulary nail has fewer complications and faster fracture healing. Elastic intramedulary nail is recommended for single pediatric femoral shaft fractures. However, external fixation is a better option for high energy injury of lower limbs, multiple trauma or severely soft tissue injury.