BACKGROUND: Compared with the bone marrow mesenchymal stem cells, umbilical cord blood mesenchymal stem cells (UCB-MSCs) is an ideal source of tissue engineered seed cells, but the culture success rate is low.OBJECTIVE: To explore establish a stable reliable method to isolate and culture UCB-MSCs by optimizing medium choice,centrifugation speed and time, incubation density, choice of growth factor, first time of medium change.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Department of Blood Transfusion, Bethune International Peace Hospital of Chinese PLA from January to October 2008.MATERIALS: A total of 20 samples of neonatal UCB by full-term uterine-incision delivery were supplied by Stem Cell Center,Bethune International Peace Hospital of Chinese PLA. Parturient and their family member signed the informed consent.METHODS: Under sterile conditions, UCB-MSCs were isolated by combination of density gradient centrifugation (1 500 r/min, totally 15 minutes) and different adherent time method. UCB-MSCs were incubated in DMEM/F12 medium, supplemented with 10% human UCB serum, 5 μg/L granulocyte-macrophage colony-stimulating factor (GM-CSF), 15 pg/L interleukin-3. MSCs at 1 ×1010/L were incubated in plastic flask coated with human UCB serum at 37℃ and 5% CO2 saturated humidity. The medium was changed following 3 days of culture. Non-adhered cells were removed. Subsequently, the medium was changed once every other 24 hours. When 80% confluence, UCB-MSCs were digested by the mixture of pancreatin-athylenediamine tetraacatic acid.MAIN OUTCOME MEASURES: Morphological changes of UCB-MSCs were observed by inverted phase contrast microscopy. Cell immunophenotypes were determined by flow cytometry.RESULTS: A small quantity of adherent round cells were determined after 24 hours, and adherent cells became more at 48 hours,with a few monopole spindle cells. Cell colonies were detected at day 7. Fibroblast-like cells arranged parallelly, presented whirlpool-shape and unclear boundary, with 80% 80% confluence 2-3 weeks following culture. At the second passage, these calls adhered at hour 12, and reached 80% 90% confluence at day 10. Flow cytometry showed that these calls were positive for CD29 and CD44, but negative for hematopoietic lineage marker CD34.CONCLUSION: MSCs can be successfully isolated from human UCB by using this modified method in vitro, with short culture cycle and high cell purification. Adherent cells have the same immunophenotype as bone marrow mesenchymal stem cells.

Objective To study the anti-neoplastic effect of Xuezhikang (Regulate blood-fat) Capsule in vitro and its influence on the chemotherapeutic drug 5 Fluorouracil (5-Fu).Methods Using MTT method to measure the inhibition rate of Xuezhikang Capsule used simply,5-Fu used simply and Xuezhikang Capsule with 5-Fu together on HepG2,MCF-7 and HL-60 cells.Results The inhibitory action of Xuezhikang Capsule(2.56,0.64,0.16,0.04,0.01 mg/ml) on HepG2,MCF-7 and HL-60 cells was dose-dependent and the IC50 was 14.057,19.859 and 27.771mg/ml respectively.The inhibition ratio of five different doses of Xuezhikang Capsule(0.64,0.16,0.04,0.01,0.0025 mg/ml) with 5-Fu (0.24mg/ml) together all remarkably higher than that of 5-Fu used simply.Conclusion Xuezhikang Capsule has a significant anti-cancer effect in vitro and can enhance remarkably the anti-neoplastic effect of 5-Fu.

AIM: To establish a sensitive method for quantitative determination of astragaloside Ⅳ (AGS-Ⅳ) in plasma and a preliminary evaluation of its pharmacokinetics parameters in intact rats. METHODS: A liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was applied for determining AGS-Ⅳ in plasma by using digoxin as the internal standard (I.S.). Six rats were given AGS-Ⅳ 2.0 mg/kg by intravenous infusion for 5 min. Blood samples were drawn intermittently with each intact rat from left femoral artery at 0.025, 0.05, 0.1, 0.25, 0.5, 1, 2, 4, 6, 10, 14 and 24 h after medication. The samples were prepared by solid phase extraction and analyzed through a triple quadrupole mass spectrometer equipped with an electrospary probe. The samples were monitored in selected ion recording (SIR) mode of positive ions by using target ions at m/z 807.5 for AS- Ⅳand at m/z 803.5 for I.S. RESULTS: Calibration curves were linear over the ranges 1-1 000 ng/mL for AGS-Ⅳ (r=0.9992). The intra-and inter-day assay variability values were less than 6% and 8%, respectively. Extraction recoveries from plasma were 92.8%-98.4% for AGS-Ⅳ and 80.0%-90.9% for digoxin, respectively. The lower limit of quantitation (LLOQ) for AGS-Ⅳ was 0.5 ng/mL. The concentration-time curves of AGS-Ⅳ for each rat were fitted to an open two-compartment model by CAPP program. The pharmacokinetics parameters of AGS-Ⅳ were as following: the elimination half-life (t1/2β), clearance rate (CL), distribution volume at steady state (Vss), and AUC0-∞ were (3.46±0.52) h, (0.47±0.02) L/h, (0.76±0.16) L/kg and (4.27±0.19) μg·mL-1·h, respectively. CONCLUSION: These results show that this method is satisfied for the measurements of pharmacokinetics study for AGS-Ⅳ.

Objective To study the correlation between genetic polymorphisms of interleukin (IL)-1A/1B and susceptibility to tuberculosis (TB).Methods Genetic polymorphisms of IL-1A and IL1 B in 1032 TB patients and 1008 non-TB patients were analyzed using PCR-MassARRAY method.The correlation between genetic polymorphisms of IL-1A/1B and susceptibility to TB was statistically analyzed.Results Two tag SNPs of IL-1A and three tag SNPs of IL-1B were screened for the study.There were differences in the allele frequencies of rs2853550 and rs3783526 between TB group and non-TB group (P=0.047and P =0.034,respectively).IL-1 B SNP1 rs2853550 (P =0.025,OR =1.302,95 ％ CI =1.034-1.640,TC vs.CC) was found to be highly associated with TB,while the other SNPs showed no significant correlations with TB.Furthermore,IL-1B SNP1 rs2853550 [P=0.019,OR=1.308,95％ CI=1.045-1.638 for (TC+TT) vs.CC] in the dominant model conferred significant risk for TB,but IL-1A SNP2 rs3783526 [P=0.000,OR=0.764,95％ CI =0.591-0.988 for GG vs.(AA+GA)] in the recessive model showed protective effects against TB.The haplotype ‘TG’ in the IL-1B block showed a higher risk for TB compared with the common ‘ CA’ haplotype (P=0.032,OR=1.265,95％ CI=1.020-1.567).The diplotypes containing ‘ GA’ haplotype in IL-1A block and ‘ TG’ haplotype in IL-1B block were major risk factors for TB (for onecopy,adjusted P=0.014,OR=1.403 and 95％ CI=1.072-1.836; adjusted P=0.013,OR=1.339 and 95％ CI=1.063-1.688,respectively),but the diplotype with ‘CG’ in IL-1B block played a protective effect against TB (for two-copy,P=0.006,OR=0.664 and95％ CI=0.494-0.891).Conclusion The genetic polymorphisms of IL-1B rs2853550 might be closely associated with TB,but the GG genotype of IL1 A SNP rs3783526 might have the characteristic of anti-TB.

OBJECTIVETo study chemical constituents of leaves from Discocleidion rufescens.

METHODColumn chromatography and spectroscapic methods were used to isolate and identify the constituents.

RESULTFifteen compounds were isolated and identified as chrysophanol (1), physcione (2), taraxerol (3), beta-sitosterol (4), daucosterol (5), scopoletin (6), apigenin (7), acacetin-7-O-beta-D-glucoside (8), apigenin-7-O-beta-D-glucoside (9 ), luteolin (10), diosmetin-7-O-beta-D-glucoside (11), luteolin-7-O-beta-D-glucoside (12), gallic acid (13), amentoflavone (14) and myo-inositol (15).

CONCLUSIONCompounds 1, 2, 8, 11, 13 and 15 were isolated from the genus Discocleidion for the first time.

Euphorbiaceae ; chemistry ; Flavones ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Sitosterols ; analysis ; isolation & purification

Objective:To explore the influence of multidimensional integration and 360° support on the function of family and marriage, and quality of life in women patients with systemic lupus erythematosus.Methods:Totally 196 patients with systemic lupus erythematosus from Department of Rheumatology and Immunology, The Second Affiliated Hospital of the Air Force Medical University from August 2016 to November 2017 were included. According to random number table method, these patients were divided into observation group and control group as 98 cases each. Conventional care and hospital discharge were used for control group. On the basis of this, multidimensional integration and 360° support were used for patients of observation group. The function of family and marriage, quality of life in patients were assessed before and after 3 months of the intervention. The treatment adherence was evaluated in 3 months and 6 months after intervention.Results:Before intervention, the marriage family function score, marital satisfaction, conflict resolution methods and the relationship with friends and family, husband and wife exchange scores of the observation group were (2.3 ± 0.5), (24.6 ± 6.1), (25.7 ± 7.1), (28.2 ± 6.9), (28.8 ± 6.9) points, respectively. Three months after intervention, these scores were (2.5 ± 0.7), (31.6 ± 5.0), (31.7 ± 5.3), (28.1 ± 6.8), (29.0 ± 7.1) points, respectively. There was statistically significant difference between before and after the intervention ( t values were -2.371 - 8.631, P < 0.01). These scores of control group before the intervention were (2.3 ± 0.6), (24.5 ± 6.2), (25.2 ± 7.2), (32.5 ± 6.0), (33.9 ± 6.3) points, respectively. Three months after intervention, these scores were (2.3 ± 0.4), (24.5 ± 6.2), (26.1 ± 6.9), (29.1± 4.8), (28.5 ± 7.2) points. Significant differeces were found between before and after the intervention in control group ( t values were -3.878-6.323, P < 0.05 or 0.01). There was statistically significant difference between the two groups after the intervention ( t values were 2.675-8.631, P<0.01). As for observation group, planning (62.8 ± 27.2 vs. 75.5 ± 25.4) and intimate relationship (62.8 ± 25.2 vs. 78.2± 24.9) in quality of life were obviously difference before and after 3 months of intervention ( t values were 3.050, 3.639, P < 0.01). As for control group, planning (62.5 ± 27.6 vs. 65.7 ± 24.9) and intimate relationship (65.8 ±25.2 vs. 63.5 ± 23.8) in quality of life were obviously difference before and after 3 months of intervention ( t values were 2.375, 3.132, P < 0.01). There was statistically significant difference between the two groups after the intervention ( t values were 3.050, 3.639, P < 0.01). The treatment adherence of observation group was significant better than control group. After 6 months intervention, the treatment adherece of observation group was 83.67% (82/98), while the treatment adherece of control group was 44.89% (44/98), significant differences were found btween the two groups ( χ2 value was 0.511, P < 0.01). Conclusion:Multidimensional integration and 360° support obviously improved function of family and marriage, improved the understanding of disease, and self-management ability of patients. Therefore, it can increase the treatment adherence and improve quality of life in SLE patients.

Humans ; Hypertension/therapy* ; Publications ; Research Design