Animals ; Dose-Response Relationship, Drug ; Humans ; Inhalation Exposure ; Mice ; Nanoparticles ; toxicity ; Rats ; Siphonaptera

Animals ; Genes, T-Cell Receptor ; genetics ; Humans ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Mice ; Mutation

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have a remarkable differentiation potential and superiority as a type of seed cells,but their application is limited in the presence of certain diseases,such as aplastic anemia and myelogenous neoplasm.The present studies have found that seed cells called muscle-derived stem cells (MDSCs) have brought more and more attention,because of their capability of stir-renewal and multi-diffcrentiation like B MSCs.OBJECTIVE: To explore the biological characterization of the muscle-derived stem cells (MDSCs) from rabbits,and analyze the phenotype.DESIGN,TIME AND SETTING: Cell in vitro observation experiment was performed at the Medical Research Center of Second Affiliated Hospital of Sun Yat-sen University from August 2005 to March 2006.MATERIALS: A New Zealand rabbit (1.5 months old,clean grade) was enrolled for the preparation of Muscle-derived stem cells.Growth medium was DMED-LG added with 10% fetal bovine serum and 10% horse serum and fusion medium was DMEM-LG added with 2% fetal bovine serum.METHODS: The muscle mass was removed from the anesthetized rabbit to isolate MDSCs.These cells were dissociated using three enzymes (collagenase XI,dispase and trypsin) respectively.Sediment was resuspended.Then preplate technique was used.The muscle cell extract was plated on a collagen-coated culture flask with growth medium.The flask was called PP1.PPI was kept overnight in a 37 ℃ incubator containing 5% CO2,After that,the suspension was transferred to another collagen-coated culture flask,which was called PP2.PP3,PP4,PP5 and PP6 were constructed later following the same procedures.The cells adhered in PP6 were collected,plated in 6-well plates,and divided into 2 groups.Growth medium was used in one group,in which the cells were kept growing at a degree of confluence beyond 50%,and fusion medium was used in the other one,in which the cells were passaged with a degree up to 30%.MAIN OUTCOME MEASURES: The cells from PP1 to PP6 were collected,and the characterization was identified preliminary by Flow cytomctry,Immunocytochemistry and Western Blotting.The fusion of cells in PP6 was detected at different confluence degrees and concentration of medium.RESULTS: The cells in PP6 showed ＞ 80% desmin+,＞ 70% Bcl-2+,＞ 95% CD45,which indicated that MDSCs were in a high concentration.The expressions of α-SMA in the cells were decreasing with the Preplate technique used and the cells in PP6 almost had no α -SMA expression.When passaged at a high confluence (＞ 50%) or cultured with low concentrations of serum (2% serum),the cells in PP6 had a strong tendency of fusing into myotubes or cell chains and were skeletal myosin+.CONCLUSION: MDSCs,which are capable of multi-differentiation under a high fusion or low serum conditions,express dcsmin and Bcl-2 highly,but extreruelv little CD45 and no α -SMA.

Objective To observe and evaluate the clinical effect of submental island myocutaneous flap(SIMF) and free skin graft(FSG) for the reconstruction of buccal defect resulted from the buccal carcinomas .Methods Forty-four aged aging patients di-agnosed with the buccal carcinomas were operated by the total dissection of primary tumor and selective neck dissection and recon-structed simultaneously with SIMF(19 cases) and FSG(25 cases) .The degree of postoperative cheek shape ,limitation of mouth o-pening ,speech sound ,and function of chewing were observed and analyzed .Results Eighteen SIMFs were completely survived .The residual muscle flap had a good blood supply after the debridement of skin island in one case ,the successful rate of flaps was 94 .7%(18/19) while 100% (25/25) .The donor site of SIMF was sutured directly .Follow-up was taken in 12 months for all patients .In SIMF group ,the degree of postoperative cheek shape ,limitation of mouth opening ,speech sound ,and function of chewing were bet-ter than those of FSG group in FSG group(P<0 .05) .There was no significant difference between 2 groups in the field of recur-rence and metastasis although the recurrence adjacent to the primary tumor had happened in FSG group in 5 months after operation (P>0 .05) .Conclusion The submental island flap is an excellent choice for the reconstruction of cheek defects in aging patients af-ter resection of buccal carcinoma .With acceptable cosmetic ,functional results and reasonable oncological saftety ,SIMF has a prom-ising prospect in head and neck surgery .

Cytokinesis ; Micronucleus Tests

Objective To summarize reasonable therapeutic measures by analyzing the characteristics of tardive intestinal malrotation in childhood. Methods Clinical data of 23 definitely diagnosed cases of intestinal mal-rotation from 1998 to 2008 were studied retrospectively. Results All of 23 patients were processed through plain ab-dominal radiograph;9 were examined with barium meal;2 were examined with barium enema;20 were examined by abdominal Doppler and 13 were examined by abdominal computer tomography. There were 18 patients who got final diagnosis preoperatively and the rate of final diagnosis was 78.3%. All of the cases were cured with Ladd's operative method. Conclusion Because tardive intestinal malrotation has various clinical manifestations,it is more difficult to diagnose than congenital intestinal malrotation in children. The main causes of death are intestinal volvulus and/or necrosis. At present,surgery is a traditional therapeutic way. Ladd procedure is the main traditional way to treat mal-trotation.

UNLABELLEDOBJECTIVE To investigate the effect of Nepsilon-(carboxymethyl) lysine albumin (CMLs), a primary advanced glycation end products (AGEPs) isoform in diabetic body, on the function and angiogenesis of adipose tissue-derived stem cells (ADSCs) and the protective effect of Danhong Injection (DH). METHODS Human ADSCs were cultured and separated from human subcutaneous fatty tissue using enzymatic digestion and centrifugation. The morphology was observed using optical microscope and differentiation capacities assessed. Cells were exposed to 5 different interventions respectively for 24 h, i.e., PBS, 60 1 microg/mL BSA, 60 microg/mL CML-BSA, 100 microL/mL DH, and 60 micro./mL CML-BSA +100 microL/mL DH. Their effect on the proliferation, migration, apoptosis, and secretion were observed using WST-1 assay, Transwell assay, Annexin V-FITC/PI flow meter test reagent kit, human VEGF reagent kit, ELISA reagent kit, respectively. The effect on ADSCs angiogenesis was observed by in vitro angiogenesis test.

RESULTSCompared with the BSA group, the capacities of proliferation and migration could be significantly inhibited by CML-BSA, the apoptosis promoted, the secretion of VEGF reduced, and the angiogenesis of ADSCs weakened (P < 0.05). Compared with the blank control group, 100 microL/mL DH could significantly promote the proliferation and migration capacities of ADSCs, inhibit apoptosis of ADSCs, increase the secretion of VEGF, and improve the angiogenesis of ADSCs (P < 0.05). Compared with the CML-BSA group, the inhibition of CML-BSA on the proliferation and migration capacities of ADSCs could be significantly reversed, the promotion of CML-BSA on the apoptosis of ADSCs improved, the secretion of VEGF increased, and the angiogenesis of ADSCs elevated (P < 0.05).

CONCLUSIONclusion CMLs could significantly inhibit the proliferation and migration capacities of ADSCs, promote their apoptosis, and inhibit their angiogeneses, which could be improved by DH.

Adipose Tissue ; cytology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glycation End Products, Advanced ; pharmacology ; Humans ; Neovascularization, Pathologic ; drug therapy ; Stem Cells ; cytology ; drug effects

A novel and sensitive HPLC-UV method has been developed for the simultaneous determination of twelve major compounds in Longdan Xiegan Pill.The chemical profile of the twelve compounds,including geniposidic acid（1）,geniposide（2）,gentiopicroside（3）,liquiritin（4）,crocin（5）,baicalin（6）,wogonoside（7）,baicalein（8）,glycyrrhizic acid（9）,wogonin（10）,oroxylin A（11）and aristolochic acid A（12）,was acquired using high-performance liquid chromatography-diode array detector coupled with an electrospray tandem mass spectrometer（HPLC-DAD-ESI-MS）.The analysis was performed on a Dikma Platisil ODS C18 column（250 mm×4.6 mm,5 μm）with a gradient solvent system of acetonitrile-0.1% aqueous formic acid.The validation was carried out and the linearities（r〉0.9996）,repeatability（RSD〈1.8%）,intra-and inter-day precision（RSD〈1.3%）,and recoveries（ranging from 96.6% to 103.4%）were acceptable.The limits of detection（LOD）of these compounds ranged from 0.29 to 4.17 ng.Aristolochic acid A,which is the toxic ingredient,was not detected in all the batches of Longdan Xiegan Pill.Furthermore,hierarchical cluster analysis was used to evaluate the variation of the herbal prescription.The proposed method is simple,effective and suitable for the quality control of this traditional Chinese medicine（TCM）.

Recently, the immunotherapy has been highlighted among cancer treatments. Cancer-testis antigen (CTA) has been studied in a variety of solid tumors because of its specific expression in tumors, and testis, ovary and placenta tissues, but not in other normal tissues. In order to provide a new approach for multiple myeloma (MM) immunotherapy, we examined the CTA expression in MM cell lines, and primary myeloma cells in patients with MM. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of MAGE-C1/CT7, SSX1, SSX2 and SSX4 in MM cell lines of RPMI-8226 and U266, and bone marrow (BM) cells of 25 MM patients and 18 healthy volunteers. The results showed that the 4 CTAs were expressed in RPMI-8226 and U266 cell lines. The positive expression rate of MAGE-C1/CT7, SSX1, SSX2 and SSX4 in the BM cells of 25 MM patients was 28% (7/25), 80% (20/25), 40% (10/25) and 68% (17/25), respectively. In contrast, the expression of any member of the CTAs was not detected in BM cells of 18 healthy volunteers. The expression of two or more CTAs was detected in 80% (20/25) MM patients, and that of at least one CTA in 88% (22/25). The mRNA expression levels of SSX1 and SSX4 were significantly higher in patients with MM at stage III than in those at stage I and II (P<0.05). No statistically significant differences were observed in the mRNA expression levels of MAGE-C1/CT7 and SSX2 in further stratified analyses by age, gender, MM types and percentage of MM cells in BM (P>0.05). In conclusion, our present study showed that MAGE-C1/CT7, SSX1, SSX2 and SSX4 were co-expressed in MM cell lines and the primary myeloma cells in MM patients, but not expressed in BM cells of healthy subjects. The mRNA levels of SSX1 and SSX4 are associated with MM clinical stage. This work may provide a new insight into MM immunotherapy in the future.

Objective To explore the relationship between the inflammatory reaction and insulin function in children with critically ill.Me-thods Ninty-six children with critical disease in Oct.2003 to Oct.2006 were enrolled in the study.Blood sugar,plasma insulin,C-peptide,tumor necrosis factor(TNF)-?,C reactive protein(CRP)were measured in the peak period and convalescence.Results Blood sugar and plasma levels of insulin,C-peptide,TNF-?,CRP were significantly higher in the peak period than those in the convalescence(Pa