BACKGROUND:The traditional two-dimensional culture system has been widely used in the in vitro culture of human tissue stem cells,but it cannot really simulate the three-dimensional physiological microenvironment in the body, which is not conducive to the study of the biological behavior of human stem cells. OBJECTIVE: To detect the effect of the bioactivity of Col-Tgel in human hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs)in vitro and in vivo,by constructing a three-dimensional culture system stimulating the physiological microenvironment of the body. METHODS:(1)In vitro co-culture:Green fluorescent protein labeled MSCs(MSCs-GFP)and human umbilical cord blood CD34+cells were co-cultured in Col-Tgel for 3 days (three-dimensional culture group). Human umbilical cord blood CD34+cells were cultured in Col-Tgel for 3 days as single culture group. MSCs-GFP and human umbilical cord blood CD34+cells were co-cultured in Transwell chamber for 3 days as two-dimensional culture group. Human umbilical cord blood CD34+cells were cultured routinely as control group. The percentage of CD34+CD38-CD45RA-CD90+cells in each group was measured by flow cytometry. In situ immunofluorescence staining was used to detect the activity of cells that were co-cultured in Col-Tgel.(2)In vivo transplantation:NOD/SCID mice subjected to 24-hour X-ray irradiation were divided into two groups: in experimental group, MSC-GFP cells were resuspended in Col-Tgel and transplanted into the tibia of NOD/SCID mice; in control group, MSCs-GFP were resuspended in PBS and transplanted into the tibia of NOD/SCID mice. The MSC-GFP growth in the bone marrow was detected by two-photon/confocal microscopy at 3 days post transplantation. RESULTS AND CONCLUSION: (1) After co-culture in Col-Tgel for 3 days, the percentage of CD34+CD38-CD45RA-CD90+cells in the three-dimensional culture group was 2.8 times that of the two-dimensional culture group, indicating that the MSCs significantly promoted the expansion of CD34+CD38-CD45RA-CD90+cells in the Col-Tgel. The percentage of CD34+CD38-CD45RA-CD90+cells in the three-dimensional culture group was increased by 4.5 times compared with the single culture group and increased by 1.5 times compared with the control group. Immunofluorescence staining showed that the cell viability of human MSCs and human umbilical cord blood CD34+cells was not affected after co-cultured in Col-Tgel for 3 days.In the in vivo transplantation experiment,MSC-GFP cells could survive in the medullary cavity.In summary, Col-Tgel provides a new strategy for stem cell culture and in vivo growth by forming a three-dimensional system similar to the physiological environment in vivo.

AIMTo investigate the effects of human urotensin II (hUII) on in vivo pia mater microcirculation in rats.

METHODSAdult SD rats were randomly assigned to the following groups: control, sodium chloride injection (NS), UII(10(-6) mol/L), noradrenaline (NA, 10(-6) mol/L), and UII (10(-6) mol/L) + NA (10(-6) mol/L) groups. For recording of microcirculation images in pia mater, skull windows were performed and mounted on the stage of an intravital microscope equipped with a TV camera. Video images of microcirculation were stored by a video cassette recorder. Temporal changes in internal diameter and microcirculatory velocity of microvessels were measured by computer using the Image Pro software. The blood flow in cerebral tissues were measured with PIMII laser Doppler perfusion Imager (Lisca, Sweden).

RESULTSThe internal diameters of arterioles and venules in control group were (35.4 +/- 3.6) microm and (40.6 +/- 8.5) microm, respectively. In UII group, the arterioles and venules contracted immediately after treated with UII and up to the peak at 1 min, the internal diameters of arterioles and venules were (25.6 +/- 3.4) microm and (23.4 +/- 3.3) microm, respectively (P < 0.05). Both microcirculatory velocity in arterioles and venules had no significant changes in UII group (P > 0.05). The blood flow in meninges increased 1 min after treated with UII and up to high peak at 5 min (3.5 +/- 0.4 perfusion unit vs. control 2.3 +/- 0.6, P < 0.05).

CONCLUSIONhUII can contract microvessels in pia mater of rats and increase microcirculatory blood perfusion to cerebral tissue involved.

Animals ; Cerebrovascular Circulation ; drug effects ; Humans ; Male ; Microcirculation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Urotensins ; pharmacology

ObjectiveTo investigate the iodine nutritional status of pregnant women,newborn heel blood thyroid stimulating hormone(TSH) level and their relationship with urinary iodine(UI) level during pregnancy in Zhoupu and Kangqiao districts of Pudong New Area of Shanghai.Methods A total of 993 urinary samples(the first,second and third trimesters of pregnancy were 200 people,respectively),breast feeding(193 people) and non-pregnant women (200 people) in Zhoupu and Kangqiao districts of Pudong New area were collected from Apr 2009 to Dec 2010.Two hundred copies of neonatal heel blood samples were collected.Median of UI was measured by arsenic-cerium catalysis.TSH in neonatal heel blood was analyzed 72 h after birth by time resolved fluoroisnmunoassay(TRFIA).ResultsMedian UI of all pregnant women was 161.35 μg/L,and that in third trimesters of pregnancy( 126.35 μg/L) was lower than that of the first,the second,the breast feeding and non-pregnant women (178.80,180.50,167.90,163.40 μg/L,all P＜ 0.05).The percentage of UI level less than 150 μg/L in the third trimester[57.5％(115/200) ] was higher than that of the first[39.0％(78/200) ],the second[39.5％(79/200) ],the breast feeding [ 16.6％ (32/193) ] and non-pregnant women [ 23.0％ (46/200) ],respectively (all P ＜ 0.05).The percentage of UI level higher than 300 μg/L in the first [9.0％(18/200)],the second[8.0％(16/200) ] and the third trimester [ 5.0％ ( 10/200 ) ] of pregnancy was lower than that of the breast feeding [ 20.2％ (39/193) ] and nonpregnant [20.5％(41/200) ] women,respectively(all P ＜ 0.05).The level of neonatal heel blood TSH was(2.92 ± 1.83)mU/L,the range was 0.01 - 9.76 mU/L,11.0％(22/200) of the neonates heel blood TSH level(5 mU/L)exceeded the ratio of World Health Organization (WHO) standard ( ＜ 3％ ) suitable for iodine nutrition.Conclusions The overall level of iodine nutrition among pregnant women in Zhoupu and Kangqiao districts of Pudong New Area of Shanghai is in the appropriate range,but the pregnant women in the third trimester is in mild iodine deficiencies,and the neonates in these districts may be prone to iodine deficiency.Monitoring of iodine nutrition of pregnant women should be strengthened and iodine supplementation should be done scientifically.

Considering the existing shortages in evaluation criteria, training systems and management modes, administrators from graduate schools of Shanghai Jiao Tong University School of Medicine (SJTUSM) performed reforms and practice for the establishment of tutor teams which should meet the requirements for culturing talented medical graduates in recent years. We have established a multidimensional evaluation criteria system oriented by both representative academic achievements and the quality of graduate students. We have strengthened horizontal cooperation with relevant administrative departments in SJTUSM. We also emphasize the concepts of "training the trainees" coordinated with administrators in the training units to implement reform measures. Herein we have summarized the main reform measures in SJTUSM to constantly improve the quality of postgraduate tutor team and guarantee high-level postgraduate education.

Objective To analyze the protein expression of costimulatory molecule B7-H1 in muscular tissues of patients with polymyositis (PM) and limb-girdle muscular dystrophy-2B type (LGMD-2B), and investigate its relevance to the pathogenesis of PM and its role in the diagnosis and identification of PM. Methods Forty-three patients with PM, 26 patients with LGMD -2B and 21 with normal muscle biopsy were recruited. Muscle biopsy was performed before frozen sections, and then, HE staining and immunohistochemistry were employed to detect the protein expression of B7-H1 in muscle tissues of each group. Results The results of HE staining of muscle tissues in the PM group and LGMD 2B group were very similar; varying degrees of necrosis, phagocytosis and regeneration phenomenon were noted with varying degrees of inflammatory cell infiltration. In PM group,muscle-related expression of B7-H1 was observed on the surface of muscle fibers (the cytomembrane). It was localized in areas where inflammatory cells lay in close apposition to damaged or non-necrotic muscle fibers. The B7-H1 protein in the PM muscular tissue was significantly increased as compared with that in the LGMD -2B tissue and normal tissue (69.77%, 26.92%, 4.76%, P<0.05). Conclusion Costimulatory molecule B7-H1 is highly expressed in the muscular tissue of patients with PM and it may be involved in the immunological pathogenesis of PM. It can be used to make a distinction between PM and other myopathies that have secondary inflammatory changes.

OBJECTIVETo investigate the association of p15 and pl6 genes deletion and STKI5 gene overexpression in primary hepatocellular carcinoma (PHC).

METHODSThe carcinoma tissue and the adjacent normal tissue were taken from 30 PHC patients during operations who had had neither chemotherapy nor radiotherapy preoperatively. DNA was extracted from the tissues and PCR was used to determine the homozygous deletion of p15 exon2 (pl5E2) and pl6 exon 2 (pl6E2). RNA was extracted, cDNA was synthesized by RT-PCR, and the expression of STKI5 gene was tested by PCR. Beta-actin was used as an internal control. Average density value (ADV) of STK15 gene and that of beta-actin gene were determined in both carcinoma tissue and the adjacent normal tissue.

RESULTSThe rate of p15E2 deletion was 13.3% (4/30) and the rate of p16E2 deletion was 16.7% (5/30) in the carcinoma tissue. The p15E2 and pl6E2 co-deletion rate was 6.7% (2/30). In 19 of the 30 cases (63.3%) the expression of STK15 gene in carcinoma tissue was higher than that in the adjacent normal tissue. The ratio of ADV of STK15 gene to ADV of beta-actin gene (1.53+/-0.31) in the carcinoma tissue was significantly higher than that (0.91+/-0.25) in the paired adjacent normal tissue (t = 2.86).

CONCLUSIONThe homozygous deletion of p15E2 and p16E2 and overexpression of STKI5 gene may play a role in the oncogenesis and malignant progression of PHC.

Aurora Kinase A ; Aurora Kinases ; Carcinoma, Hepatocellular ; genetics ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Female ; Gene Deletion ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; genetics ; Male ; Protein-Serine-Threonine Kinases ; biosynthesis ; genetics

AIMTo establish an HPLC-fluorescent spectrometric method for the determination of mexiletine hydrochloride in plasma after derivatization with fluram.

METHODSFluram acetone solution was added to the deproteinized plasma with acetone to obtain the derivative of mexiletine. The HPLC method was performed on a column of Allitima C18 (150 mm x 4.6 mm, 5 microns) with the mobile phase of methanol-water-diethylamine-phosphoric acid buffer (2.4 mol.L-1, pH 4.0) (70:28:2), and the detective wavelength were set at Ex 392 nm and Em 480 nm.

RESULTSMexiletine has a liner range over the concentration range from 0.100-6.400 mg.L-1. The lowest detectable concentration of this method was 5 micrograms.L-1 (S/N > or = 4). The intra-day and inter-day RSDs were 1.34%-5.31%, respectively.

CONCLUSIONThis method is simple, selective and can be used for therapeutic drug monitoring (TDM) and pharmacokinetic studies of mexiletine.

Anti-Arrhythmia Agents ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; methods ; Fluorescamine ; chemistry ; Humans ; Mexiletine ; blood ; pharmacokinetics

Serum pharmacochemistry of traditional Chinese medicine (TCM) is designed to screen the efficacy material base of TCMs from the constituents absorbed into the blood after oral administration. The theory and method is in accordance with the effect characteristics of TCMs, and reflects the interaction between the body and the drugs, has become an effective pathway for researching the efficacy material base of TCMs which has been recognized and used widely. In the paper, the previous research contents and methods of the serum pharmacochemistry of TCM were reviewed, and on the basis of the further validity of the special administration form of the TCM formula and the corresponding property to TCM syndrome, the new strategy of serum pharmacochemistry of TCM integrating the metabonomics technologies was put forward. According to the strategy, we take the biological characters of TCM syndrome as a research starting point, taking TCM formula as object, using the metabolic biomarkers of syndromes or disease to evaluate the therapeutic effect of formula and screen the compounds of TCMs in serum which are highly correlated with the metabolic biomarkers through the correlation analysis, and by further biological validation to finally confirm the efficacy material basis of TCMs. Integrating with the systems biology technologies, the theory and method of serum pharmacochemistry of TCM will further develop, and open a new chapter in the interpretation of the theory of TCM.
Animals ; Drug Therapy ; trends ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Humans ; Metabolomics ; Serum ; chemistry

OBJECTIVETo study the metabolic chemistry and pharmaco-dynamics characters of ligan from seeds of Arctium lappa.

METHODHPLC method was used in the study. The analysis was carried out on C18 column. The mobile phase was CH3CN-0.05% H3PO4 (36:64) with flow-rate at 0.6 mL x min(-1) and wave-length of 210 nm. The column temperature was kept at 25 degrees C.

RESULTThe results indicated that the ligan was detected in plasma and the main organs 5 min after po. The main metabolic production in plasma was arctigenin. In addition, arctigenin and an unknown product were found in metabolic production in the organs.

CONCLUSIONThe method was stable,simple and reproducible. It can be used to determine the metabolic product of the ligan. The metabolic chemistry of ligan in plasma was obviously different from that in the main organs.

Animals ; Arctium ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Furans ; blood ; metabolism ; Glucosides ; blood ; metabolism ; Lignans ; blood ; isolation & purification ; metabolism ; pharmacokinetics ; Liver ; metabolism ; Male ; Mice ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Seeds ; chemistry ; Tissue Distribution

OBJECTIVETo analyze and identify fatty acids in the seeds of Sterculia lychnophora.

METHODThe compositions was isolated and determined by GC-MS technique, and area normalization method was used to make quantitative analyze of the content of compositions.

RESULTS21 Fatty acids and 5 other compositions were isolated and determined.

CONCLUSIONThe major fatty acids are 9,12(Z,Z)-octadecadienoic acid(37.96%), hexadecanoic acid(24.77%), 9-(Z)-octadecenoic acid(19.77%) and octadecanoic acid(5.01%).

Fatty Acids, Nonesterified ; chemistry ; isolation & purification ; Fatty Acids, Unsaturated ; analysis ; Gas Chromatography-Mass Spectrometry ; Palmitic Acid ; analysis ; Plants, Medicinal ; chemistry ; Seeds ; chemistry ; Sterculia ; chemistry