To elucidate the relationship between WT1 expression and differentiation of leukemia cells and the role of WT1 gene in differentiation of leukemia cells, HL 60 and K562 cell lines were induced for 5 days by all trans retinoic acid(ATRA).Then the degree of differentiation and WT1 expression of cell lines were determined by NBT reduction assay and RT PCR respectively. The resultsshowed that only differentiation of HL 60 cells but no K562 cells could be induced by ATRA. When HL 60 cells were induced to differentiate to granulocytes by ATRA, the expression of WT1 decreased markedly during differentiation. However, WT1 transcripts were not significantly altered in K562 cells in which differentiation wasn't found. It suggested that WT1 gene expression may relate to the differentiation of leukemia cells.

Objective:To eastablish the efficient presentation of antigen from apoptotic cells by human DC from peripheral blood. Methods: using recombinant human granulocyte/macrophage colonystimulating factor(GM- CSF) and interleukin 4 (IL- 4 ) we have established dendritic cells (DC)from peripheral blood monocyte that maintain the antigen capturing and processing capacity characteristic of immature dendritic cells in vivo. GM - CSF 50ng/ml , IL- 41 000ng/ml once two days(total four). on the 3 rd day of culture, immature DC and apoptotic hepatochlangioma cells were in coculture lasting 7 days. Results:these cells had typical dendritic morphology, express high levels of CD1a ,B7 and acquired antigen from apoptotic cells and induced an increased T cell stimulatory capacity in MLR. Conclusions:we have established DC from blood mononuclear tells using GM- CSF and IL- 4 and DC can be efficiently drived from apoptotic cells and can induce the increase of T cells obviously. It probably becomes an effective approach of antigen transduced with DC.

BACKGROUND: Dendritic cells play an important role in antigen present in vivo, and the mechanism of tumor cells in escaping the antigen presentation of dendritic cells existed in the patients with tumor.OBJECTIVE: To sensitize dendritic cells from human peripheral blood with apoptotic hepatoma cells induced by mitomycin.DESIGN: A randomized control trial by taking apoptotic hepatoma cell sensitized dendritic cells as the observed subjects.SETTINGS: Institute of Field Surgery, Daping Hospital, the Third Military Medical University of Chinese PLA; Institute of Radiation Medicine,Chinese PLA Academy of Military Medical Sciences.MATERIALS: The experiments were carried out in the Institute of Radiation Medicine, Chinese PLA Academy of Military Medical Sciences from April 1998 to May 1999. The cell strain was the QBC939 bile duct cancer cell strain, and mitomycin was used as the antitumor drug.METHODS: Mononuclear cells were isolated from the peripheral blood of normal people, 50μg/L granulocyte-macrophage colony stimulating factor(GM-CSF) and 1 000 U/mL interleukin-4 (IL-4) were added, once every other day for 4 times. On the 3rd day of culture, the apoptotic bile duct cancer cells induced by mitomycin was added, and then cultured in vitro for 4 days, finally the dendritic cells were collected.MAIN OUTCOME MEASURES: ① The identification of the cultured dendritic cells was observed; ② The dendritic cells were co-cultured with necrotic and normally cultured bile duct cancer cells respectively, and the phagocytized apoptotic body loaded antigens were observed; ③ The immunostimulatory activity of dendritic cells (1×103, 5×103 and 1×104/well)and that after loaded by antigen were detected, and the mononuclear cells were taken as controls.RESULTS: ① The cultured and amplified dendritic cells expressed high levels of costimulatory molecules of CD1a and B7, and there were typical irregular processes on the surface. ② The tumor cells formed apoptotic bodies when they were induced by mitomycin, which were arrested and phagocytized by dendritic cells. ③ The ability of the antigen loaded dendritic cells in stimulating the proliferation of allogenic lymphocyte T was further enhanced.CONCLUSION: The apoptotic tumor cells induced by mitomycin can induce the mononuclear cells from human peripheral blood differentiating into the dendritic cells with the concomitance of GM-CSF and recombinant IL-4 and amplify dendritic cells. Meanwhile, the dendritic cells can effectively present the antigens of apoptotic bile duct cancer cells, and it probably becomes a new effective approach for tumor antigen to sensitize dendritic cells.

Objective To observe the value of dynamic contrast-enhanced MR angiography (DCE MRA) with gadobenate dimeglumine in evaluation of vascular complications after living donor liver transplantation. Methods Thirty-four consecutive patients were examined with MR after living donor liver transplantation. First, 1 ml gadobenate dimeglumine was injected in vein to infer the time of gadobenate dimeglumine reaching abdominal artery performing testbolus. Then a scan with three-dimensional T1-weighted fast low-angle shot (3D-FLASH) sequence was performed. Four phases in series from arterial period were scanned and every phase paused 10 s to obtain images of the arterial, portal venous and hepatic venous systems. The original and maximum intensity projection (MIP) reconstructed images, categorized vessel visualization on a five-point scale and observed stoma of hepatic artery, portal venous and hepatic venous inferior vena cava, diameter and display of surrounding vessels were observed. The results were compared with those of digital subtraction angiography (DSA), ultrasound and clinical data. Results Overall vessel visualization assessment demonstrated good or very good ratings for the majority of patients. Among all 34 patients, hepatic artery stenosis was found in 4, portal vein stenosis in 6, portal vein thrombosis was detected in 2, while middle hepatic veins stenosis was detected in 1 patient, among whom 10 patients were confirmed with DSA, 4 with surgery, the others with ultrasound or follow-up. Conclusion Gadobenate dimeglumine DCE MRA is a highly accurate, noninvasive tool for evaluation of vascular complications after living donor liver transplantation, may be regarded as the first choice in postoperative evaluation.

Objective To investigate the feasibility of measuring liver volume with Argus methoct Methods Thirty-two healthy liver transplant donor candidates underwent liver MRI on a 3.0 T MR unit.Volume interpolated body examination(VIBE)was performed after the administration of gadobenate dimeglumine.The VIBE data was transferred to the diagnostic workstation,and then multiple planar reconstruction(MPR)images were acquired.Firstly.two observers manually drawn the liver shape and calculated three volumes:the whole liver volume and right lobes volumes include middle hepatic vein (MHV)and exclude MHV,respectively.Secondly,the same data was transferred to Argus software.calculated that three volumes.Each measurement time was recorded.Actual graft volume(the right lobe)wag measured during surgery.The correlation between right lobes volume of two measurements and actual graft volume was analyzed.The time needed for Argus and that needed for manual method were compared with paired t test.Results The right lobe volumes measured by Argus,manually and surgery method were (813±187),(807 ± 181)and(713 ± 137)mm3,respectively.Argus method and manual method showed good correlation with surgery method,and the correlation coefficients were 0.897(Argus method)and 0.884(manual method),respectively.The time for manual method and Argu8 method were(44.3 ±2.7)and(12.2.±1.0)min,respectively.There was significant difference between Argus and manual methods (t=76.39,P＜0.05).Conclusion Compared with manual method,use of the Liver volumetric measurement by Argus software not only correlated well with Actual graft volume,but also saves time.Argus has potential clinical value for volumetric measurement in living liver transplant donors.

Objective To compare conventional T2-weighted MR cholangiography (T2WI-MRC) with gadobenate dimeglumine enhanced T1-weighted MR cholangiography(CE-MRC) for evalution of biliary anatomy in liver transplant donor candidates. Methods Thirty-two healthy liver transplant donor candidates were examined with two MR cholangiogaphic methods. For T2WI-MRC, a three-dimensional turbo spin-echo sequence and oblique coronal heavily T2-weighted thick-slab turbo spin-echo imaging sequence were performed. For CE-MRC, three-dimensional fat-suppressed spoiled gradient-echo sequences were performed, with a time delay of 60 minutes following the administration of gadobenate dimeglumine. To compare the depiction of biliary duct anatomy and the artifact caused by intestinal liquid and breathing between the two methods. Intraoperative cholangiography was the reference-standard examination. Results The both methods depicted the biliary anatomy correctly in all 9 cases. The both methods showed the third branches of intrahepatic biliary duct clearly. T2WI-MRC showed interhepatic bihary duct before the third branches in 28 cases (87.5%), CE-MRC showed the same finding in 14 cases (43.8% ). T2WI-MRC showed common bile ducts intermitantly in 2 cases, which were normal in CE-MRC and intraoperative cholangiography. Intestinal liquid affected the image quality of biliary duct in 6 cases (18.8%) performed with T2WI-MRC, but none with CE-MRC. The artifacts caused by breathing were not obvious in the either method. Conclusion T2WI-MRC and CE-MRC both can be used to evaluate bihary anatomy of liver transplant donor candidates, but CE-MRC appears to be more accurate than T2WI-MRC.

Objective To explore whether dynamic contrast-enhanced MRA (DCE MRA) can provide an effective assessment of renal vascular in living donors before transplantation.Methods Thirty five healthy living renal donor candidates were scanned on MR system before transplantation.After injection of Gd-DTPA 1 ml in vein, a test-bolus scan was used to get the time delay of Gd-DTPA reaching renal artery.Then, a 3D T1-weighted fast low-angle shot sequence (3D FLASH) was performed in the coronal plane.The 3D FLASH scan would repeat four times with an inter-phase of 10 seconds.Thus, the imaging of the renal arterial, venous and collecting systems were got.Two radiologists observed renal arteries and veins on original imaging and MIP reconstructed imaging.The quality of MR angiography was evaluated on a fivepoint scale and the vascular anatomy or variations of the arterial and venous systems were recorded, using intraoperative findings as a standard of reference.Results The quality for all MRA was good or very good for the most of living renal donors.Among 70 renals, several variations of vascular were found, including 5 left accessory artery, 9 right accessory artery, 3 left proximal arterial branch and 6 right proximal arterial branch.Among 70 renal veins, 1 right accessory veins and 2 left varieocele were observed.One small accessory artery of right kidney was missed with DCE MRA, but identified by operation.Conclusion DCE MRA was noninvasive tool for evaluation of the renal vasculature and variations with high accuracy.It would be a good modality in preoperative evaluation of living renal donors.

Objective:To estimate the pharmacokinetics for two solution types of propofol glycoside in-jections in rats .Methods:A high performance liquid chromatography-high resolution mass spectrometry ( HPLC-MS) was established for measuring propofol in rat plasma .Two kinds of propofol glycoside injec-tions were developed and intravenously administered to rats via tail vein , respectively , and a commercial-ly available propofol emulsion injection was intravenously administered as a control .Propofol plasma concentration-time curves were determined , and the pharmacokinetic parameters were estimated .Re-sults:HPLC-MS measurement was performed by using a quadrupole-orbit trap high-resolution mass spec-trometer on a C18 chromatographic column.The mobile phase consisted of water and methanol (20∶80, V/V) .The ion source was an atmospheric pressure chemical ion source , and the negative ion was used for detection with a scanning mode of selective ion monitoring in which m/z 177.127 4 was used for propofol and m/z 149.096 1 used for thymol as an internal standard .A linear correlation between con-centration and peak area ratio was constructed in the range of 50 μg/L-10.0 mg/L propofol.The limit of quantification was 50μg/L propofol .The average recoveries of propofol from plasma were in the range of 93.6% -101.1%, and intra-day or inter-day relative standard deviation for measurement was <14%.The pharmacokinetic results showed that the two kinds of propofol glycoside injections exhibited the same pharmacokinetic behavior .However, the clearance and area under curve values of propofol for the two propofol glycoside injections were evidently increased as compared with those for propofol emulsion injection, respectively.Furthermore, their apparent distribution volumes were increased as well .Never-theless, the propofol elimination half-life (t1/2) value of the newly developed propofol glycoside injections was the same as that of commercial propofol emulsion injection (approximately 1.5 h).Conclusion:The established HPLC-MS method can be used for measuring propofol concentration accurately in rat plasma . The clearance and distribution volumes of propofol glycoside injection are bigger than those of the propofol emulsion injection .

OBJECTIVE Study the role of estrogen receptor (ER)in the inhibition of cell viability and differentiation induced by bisphenol A (BPA)in micro mass culture of rat e mbryonic midbrain(MB) cells.METHODS Micro mass cultures of MB were prepared fro m rat e mbryonic midbrain on gestation day 13.MB cells were exposed to BPA (10 -4 ,10 -6 ,10 -8 ,10 -10 ,10 -12 mol·L -1 )for 5 d.Cell viability was assessed by neutral red uptake test.MB differentiation was detected by he matoxylin staining and i mage analysis.In order to observe the role of ER pathway in the toxicity induced by BPA,cell cultures were co-treated with ICI182780 0.1 n mol·L -1 ,ta moxifen 1 n mol·L -1 and BPA 0.1 mmol·L -1 for 5 d, the cell viability and foci differentiation were detected.Moreover,the protein expression levels of ER in normal e mbryonic brain of gestation day 18,testis tissue fro m adult rats and midbrain cells untreated with BPA were investigated by Western blot.The mRNA expression levels of ER in normal e mbryonic brain of gestation day 13 and gestation day 18,ovary and testis tissue fro m adult rats,and midbrain cells un-treated with BPA were investigated by real-ti me PCR.The mRNA expression levels of Notch1 and Hes1 in MB cells treated with BPA 0.1 mmol·L -1 were also detected by real-ti me PCR.RESULTS BPA 0.1 mmol·L -1 could inhibited MB cell viability and foci differentiation.However,this effect could not be reversed by ER antagonist.The protein and mRNA expression levels of ER in e mbryonic brain and MB cells untreated with BPA were found to be extre mly low.In addition,BPA 0.1 mmol·L -1 could inhibited the mRNA expression levels of Notch1 and Hes1 .CONCLUSION BPA could inhibited MB cell viability and foci differentiation.ER pathway might be not involved in this effect.Instead,Notch-Hes pathway might be involved for this effect.

Objective To evaluate the effect of peritoneal resuscitation (PR) with pyruvate-based peritoneal dialysis solution (PDS) on kidney injury in a rat model of hemorrhagic shock.Methods Fifty healthy adult male Sprague-Dawley rats,weighing 200-250 g,aged 8 weeks,were divided into 5 groups (n=10 each) using a random number table:sham operation group (group SH),conventional Ⅳ resuscitation group (group VR),PR with normal saline group (group NS),PR with lactate-based PDS group (group LA) and PR with pyruvate-based PDS group (group PY).Hemorrhagic shock was induced by withdrawing blood from the left femoral artery at a rate of 0.6 ml/min about 10 min until mean arterial pressure was reduced to 30-40 mmHg which was maintained for 1 h.In group VR,the animals were resuscitated with infusion of the blood withdrawn and normal saline (the volume was 2 times volume of blood loss) at 1 h after hemorrhagic shock.In NS,LA and PY groups,conventional Ⅳ resuscitation was performed,and the animals were simultaneously resuscitated with normal saline,lactate-based PDS,and pyruvatebased PDS 20 ml infused intraperitoneally over 30 min,respectively.The animals were sacrificed at 180 min after resuscitation,and kidneys were removed for examination of the pathological changes (with a light microscope) and for measurement of the content of malondialdehyde (MDA) and activities of myeloperoxidase (MPO) and superoxide dismutase (SOD) in renal tissues.The damage to renal tubules was assessed and scored.Results Compared with group SH,the renal tubular damage scores,MDA content and MPO activity were significantly increased,and the activity of SOD was decreased in the other four groups (P＜0.05).Compared with group VR,the renal tubular damage scores,MDA content and MPO activity were significantly decreased,and the activity of SOD was increased in NS,LA and PY groups (P＜0.05).Compared with group NS or group LA,the renal tubular damage scores,MDA content and MPO activity were significantly decreased,and the activity of SOD was increased in group PY (P＜0.05).The pathological changes of renal tissues were significantly attenuated in group PY when compared with VR,NS and LA groups.Conclusion PR with pyruvate-based PDS can reduce kidney injury in a rat model of hemorrhagic shock.