[Objective]To investigate if allogeneic human serum(alloHS)could replace fetal bovine serum(FBS)for the expansion of human bone marrow mesenchymal stem cells(hBMSCs)in vitro.[Method]Human BM-MSCs were isolated either from spongy bone residues obtained during hip surgery or from washed filters used during bone marrow graft processing for allogeneic bone marrow transplantation.We culture human bone marrow mesenchymal stem cells were cultivated using alloHS and FBS,then observe its growth character,antigen presentation and differentiation potency through condition medium directional induction were observed.[Result]Isolated MSCs were positive for the markers CD105,CD29 and CD44 and negative for typical hematopoietic and endothelial markers CD34,CD45,CD106 and HLA-DR.The choice of serum affected hMSCs at several different levels.Firstly,hMSCs in alloHS proliferated markedly faster than hMSCs in FBS.Secondly,hMSCs in FBS differentiated more rapidly toward mesenchymal lineages compared with hMSCs in alloHS.[Conclusion]hMSCs may be expanded rapidly in alloHS in the absence of growth factors,this may be a safely culture method suitable for clinical demand.

A Comprehensive review was present on the sources, enzyme stru cture, enzyme reaction mechanism and the application of the nitrilase.

Purpose:To investigate the effectiveness of intralymphatic infusion of anti- cancer agents and cytokines in the treatmrnt of malignancy.Materials anti methods:23 patients suffering from advanced metastatic cancers anti 2 primary lymphomas,unresponsble to the standard therapies or intra-arterial chemotherapy,were treated with lymphatic injections of an- ticancer ddrugs or combiation with biochemotherapy.Results:Follow-up study about one month after the therapy,comparing with findings on lymthatic radiographies anti computed tomographic scans,revealed decrease of lymphnodes in size in 23 cases.Conclusion:This therapeutic ap- proach proved to be an effective and safe method for the palliative treatment of advanced lym- phatic metastases and lymphomas.The procedure was feasible without serious compllications.

Objective To investigate the biological function of miR-622 in human gastric cancer cell lines of SGC-7901 and NCI-N87 cells and its role in gastric carcinogenesis.Methods We analyzed the expression of miR-622 in those human gastric cancer cell lines by quantitative real-time polymerase chain reaction.Tumorigenesis,migration and invasion ability of miR-622 overexpression was assessed in vitro with miR-622 precursor and inhibitor in in SGC-7901 and NCI-N87 cells.Results The expression level of miR-622 in SGC 7901 was 1.29 ± 0.57,and it was 10.96 ± 1.02 in NCI-N87 cells.The soft agar colony formation rate was 76％ in SGC-7901 after transfecting miR-331-3p precursor,the ability of scratch healing was ( 11 ±7) μm,and the ability of transwell invasion was (731 ±3),compared with that in control group,the differences were statistically significant ( P ＜ 0.05 ).Conclusions Over-expression of miR-622 promotes tumorigenesis,migration,and invasion in gastric cancer cells in vitro.

Purpose To build up the Potassium Sodium Dehydroandroan Drographolide Succinate enteric coating sustained-release pellets delivery system by aqueous dispersion coating technique. Methods 1. Adopting MCC as vehicle, SiO_2 as antisticking agent, appropriate amount of 40% ethanol, preparing the core of pellets by extrude-spheronization method and the formulation and manufacturing process were optimized by orthogonal design. 2. Preparing the coating material with Eudragit L 30 D, which is used as pore-forming agent, EC as blocker and PEG6000 as plasticizer. The pellets were coated by fluid-bed coating method. Results 1. The optimal formulation and manufacturing process of pelltes' core were as follows: drug: MCC: SiO_2 = 7:7:5, extruding rate: 1 080 r/min, rounding rate: 960 r/min, rounding time: 5 min. 2. After the addition of EC: Eudragit L = 35:65, the plasticizer is 1.71 % and weight gain is 5% . The release in the gastric model fluid(pH 1.0) < 10% , and complete release ( > 80% ) in the enteric model fluid(pH 6.8) was in two hours. The release behavior accords with the regulations on the release rate of enteric preparation in ChP. Conclusion By adjusting the formulation and the parameters of the process of pellet and coating, we can make enteric coating Potassium Sodium Dehydroandroan Drographolide Succinate sustained-release pellets. All this accords with the regulation of pharmacopedia in vitro release.

Objective To determine chromosomal localization of the primary gout susceptibility gene in a pedigree.Methods The clinical data and the peripheral blood samples were collected in the pedigree members and the genomic DNA was extracted from peripheral blood.A genome-wide screening was performed using 400 micro-satellite DNA markers in this family,and linkage analysis was used to determine the chromosomal location of the primary gout susceptibility gene.Results Linkage analysis showed that the maximum LOD score reached 1.50 at marker D4S1572 (at recombination fraction?=0.00).Conclusion Since D4S1572 is localized at 4q25,the primary gout susceptibility gene of this pedigree is localized at 4q25.

Objective To analyze the correlation factors between CT imaging features of pulmonary embolism(PE)and clinical severity stratification,to explore the value of CT pulmonary angiography (CTPA)in acute PE severity stratification.Methods According to the clinical severity,48 patients with acute PE proved by CTPA were classified into two groups,including 21 critical and 27 non-critical patients. Embolism index,ratio of central pulmonary involvement,ratio of right ventricle maximum minor axis (RVMMA)to left ventricle maximum minor axis(LVMMA),namely RV:LV,dilation of main pulmonary and/or right pulmonary trunk,and dilation of bronchial arteries in both groups were analyzed comparatively. The correlation factors between CT imaging features and PE clinical severity stratification were explored.The correlation between RV:LV and embolism index of 48 patients was analyzed.Results Pulmonary embolism index(22.0%—85.0%,median 38.0%),ratio of central pulmonary involvement(42.5%),RV:LV (0.90—1.90,median 1.30),dilation of pulmonary artery(14 cases),and dilation of bronchial artery (8 cases)in critical group(21 cases)were higher than those corresponding factors(5%—48%,median 21.5%,31.25%,0.80—1.40,median 1.00,5 eases,and 3 eases)in non-critical group(27 cases) (Z=4.27,X~2=5.40,Z=2.58,X~2=11.45,X~2=4.87,P

Objective To investigate the effect of angiotensinⅡ(AngⅡ) type 1 receptor blocker losartan on the cyclooxygenase 2 (COX-2) expression in metabolic syndrome (MS)kidney. Methods Seven-week-old male obese Zucker rats,a model of MS,were randomly divided into losartan treated and untreated group,and lean Zucker rats were used as controls.The obese Zucker rata of treated group received losartan for 4 months continuously.COX-2 expression was examined for all rats after 4 months.AngⅡ-stimulated mesangial cells and cortical tissue from AngⅡ-infused C57BL/6 mouse kidney by osmotic minipumps were used in this study.RNA and protein were obtained from renal cortical tissue or mesangial cells for RT-PCR and Western blot.Results Compared to the lean controls,obese Zucker rats showed a significant increase of COX-2 expression in the renal cortical tissue and these abnormalities were prevented by administration of losartan. Furthermore,the direct stimulation of AngⅡincreased COX-2 expression in mesangial cells in vitro and renal cortical tissue in vivo.Conclusions MS-induced COX-2 expression in the kidney is regulated by AngⅡ.Losartan as a non COX-2 inhibitor can protect MS kidney,at least in part,by inhibition of COX-2 activation.

OBJECTIVETo investigate whether TRAIL can synergize with adriamycin (ADM) to kill osteosarcoma cells (U2OS) in vitro, and its possible molecular mechanism.

METHODSMTT was used to evaluate the cytotoxic effect of TRAIL and ADM either used alone or in combination at 24 hours after treatment to U20S cells. Cell apoptosis and its proportion were detected by flow cytometry assay. Acridine orange fluorescence microscopy and transmission electron microscopy were used to examine cellular and ultrastructural changes of apoptosis. The changes of cFLIP in mRNA and protein level were semi-quantified by RT-PCR and Western blot analysis.

RESULTS(1) U2OS cells were not sensitive to TRAIL (IC(50) > 1 mg/L); the cells were relatively more responsive to ADM in an apparent dose-effect fashion. (2) The combination of TRAIL and ADM presented a synergistic effect on U2OS cells. Subtoxic concentration of TRAIL (0.1 mg/L) combined with subtoxic concentration of ADM (1.0 micromol/l) killed (49.54 +/- 2.79)% of U2OS cells. (3) The cytotoxicity was mainly attributed to cell apoptosis as demonstrated by flow cytometry assay, fluorescence microscopy and electron microscopy.

CONCLUSIONSubtoxic dose of TRAIL can effectively kill osteosarcoma cells (U2OS) in combination with subtoxic dose of ADM, but not effective when used alone. Apoptosis is the main mechanism of this killing effect induced by combination of TRAIL and ADM. Down regulation of cFLIP at mRNA and protein level is involved in this apoptosis pathway.

Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; pharmacology ; Bone Neoplasms ; pathology ; CASP8 and FADD-Like Apoptosis Regulating Protein ; Caspase Inhibitors ; Doxorubicin ; pharmacology ; Drug Synergism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Membrane Glycoproteins ; pharmacology ; Osteosarcoma ; pathology ; RNA, Messenger ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo identify a new allele by analyzing the polymorphism of D18S1364 in Power Plex(TM)16.

METHODSAn abnormal band was found in paternity test by short tandem repeat-PCR, and collected by gel extraction. Then the DNA was amplified, cloned and sequenced.

RESULTSA fragment containing eight bases less than the minimal allele in the D18S1364 ladder, was found with the core sequence of (ATCT)₆(ATGT)₁(ATCT)₃. The allele was D18S1364-10.

CONCLUSIONThe D18S1364-10 allele was found and reported for the first time in China.

Adult ; Alleles ; Base Sequence ; Child ; Female ; Humans ; Microsatellite Repeats ; genetics ; Molecular Sequence Data ; Sequence Analysis, DNA ; methods