Despite a combination of all available treatment, the mortality of liver failure is very high,especially in children patients. Artificial liver support methods have been tested for over 50 years. Standard techniques of blood purification like hemodialysis, adsorption, hemo or plasma filtration as well as bioreactorbased approaches using liver cells or tissues have been used. It' s believed that the damaged liver has the ability to return to normal. Artificial liver support systems are expected to be useful for temporary support of liver function. If the liver does not regenerate to normal functions, an artificial liver support system may be useful as a bridge to liver transplantation. In conclusion, artificial liver support method appears to be a reliable therapy for advanced liver diseases and has significantly decreased the mortality of liver failure. Artificial liver support system has been used in children patients as well, but it still needs more researches.

Angiotensin I ; Hepatic Stellate Cells ; Insulin-Like Growth Factor Binding Protein 2 ; Peptide Fragments ; Peptidyl-Dipeptidase A

Objective To explore the influence of cryopreservation by ladder-style freezing from low temperature refrigerator to liquid nitrogen on hemopoietic stem cell(HSC)transplantation.Methods From April 2001 to April 2006,31 cases treated with HSC transplantation were randomized to controlled-rate freezer-liquid nitrogen group(n=15)and refrigerator-liquid nitrogen group(n=16)in Department of Hematology of 1st People's Hospital of Yunnan Province.The hemopoietic reconstitution time was observed,and the viability of HSC was determined by trypan blue exclusion test in two groups.Results No significant differences were found in transfusion values of mononuclear cells(MNC)and CD34+ cells,rates of trypan blue exclusion and time of ANC0.05).Conclusion Cryopreservation of HSC can produce successful engraftment by ladder-style freezing which has cryoprotectant solution containing final concentrations of 3-percent hydroxyethyl starch,4-percent human serum albumin,and 5-percent DMSO,and its effect is the same as traditional controlled-rate method with 10-percent DMSO cryoprotectant.The new freezing procedure is faster and easier,and freezing cost is reduced.

Objective To construct rat calcineurin catalytic subunit ?(CnA)/enhanced green fluorescent protein EGFP gene coexpression plasmid for exploring the effect of calcineurin on the myocardium apoptosis induced by ischemia-reperfusion.Methods Total RNA was isolated from the heart of the adult Wistar rat,and CnA CDS segment of approximate 1 590 bp size was amplified by reverse transcription PCR method.CnA cDNA segment was cloned into pMD18-T Simple vector for sequencing,and the right clone was named T-CnA.CnA cDNA segment excised from plasmid T-CnA was ligated with pShuttle2 which had inserted IRES-EGFP segment into before.The DNA of the recombinant plasmid was extracted and was identified by double digesting with Nhe Ⅰ and Not Ⅰ.The right clone was named pShuttle2-CnA-IRES-EGFP.Results Sequencing result verified that the PCR product of CnA gene was identical to GenBank(NM_017041).1% agarose electrophoresis showed the bands of recombinant plasmid pShuttle2-CnA-IRES-EGFP digested by Nhe Ⅰ and Not Ⅰ were in the right range corresponding with expectation(1 590 bp and 5 240 bp).Conclusion Recombinant eukaryotic expression plasmid carrying rat CnA cDNA as well as a report gene-EGFP gene is successfully constructed in this experiment.

Objective To investigate the effects of wortmannin (WT) on leukemia cells cycle and BCL-2 protein expression. Methods 0 25?mol/L PI-3 kinase inhibitor wortmanin was used to treat K562 cells for 24 hours, and Arac was simultaneously used to induce the cell apoptosis. The specific fluoresent staining and FCM analysis were adopted to measure the cell cycle distribution and BCL-2 expression. Results Compared with the control cells, the proliferation index(PI) of the K562 cells treated by wortmannin was significantly lower, the cell number of G1 phase and the percentage of cell apoptosis increased, and the BCL-2 protein expression significantly decreased. Conclusion Wortmannin could inhibitedtheproliferationofK5 6 2cells ,andwascellcyclespecificagent (CCSA) .

Objective To investigate the the expression of soluble Fas (sFas) in the placenta of pregnancy induced hypertension (PIH) patients after perinatal. Methods Expression of sFas were detected by Fluorescent MGB Probe Real-Time PCR in 34 severe PIH patients and 30 healthy pregnant women served as normal controls.Results Expression of placenta sFas in 34 patients were significantly higher than those in normal controls.Conclusion PIH patients' placenta had higher expressiom of sFas; Detection of sFas may be helpful to value PIH degrade and sFas would be become a indicative markers of cell proliferation and apoptosis during the perinatal period.

Rat calcineurin (CaN) A alpha isoform (Ppp3ca) cDNA recombinant adenovirus vector was constructed in order to explore the effect of CaN on the myocardium apoptosis induced by ischemia-reperfusion injury. Total RNA was isolated from the heart of the adult Wistar rat, and Ppp3ca CDS segment of approximate 1.59 kb size was amplified by reverse transcriptional PCR method. Ppp3ca cDNA segment was cloned into pMD18-T Simple vector for sequencing, and the right clone was named T-Ppp3ca. Ppp3ca cDNA segment obtained from T-Ppp3ca was ligated with pShuttle2-IRES-EGFP to construct a recombinant plasmid pShuttle2-Ppp3ca-IRES-EGFP. Ppp3ca-IRES-EGFP expression cassette containing CMV, Ppp3ca-IRES-EGFP and SV40 polyA DNA fragment (3.97 kb) obtained from pShuttle2-Ppp3ca-IRES-EGFP was connected with pAdeno-X backbone sequence to construct a recombinant plasmid pAdeno-Ppp3ca. After being identified by PCR and enzyme digestion, recombinant plasmid pAdeno-Ppp3ca was packaged in HEK293 cells. Supernatant of adenovirus from HEK293 cells was collected after a visible cytopathic effect (CPE) appeared. The DNA of the recombinant adenovirus was extracted with the standard method. The presence of the recombinant adenovirus was verified by PCR. The results showed that sequencing results verified that the PCR product of Ppp3ca gene was identical to GenBank. Agarose electrophoresis showed the bands of recombined plasmid pAdeno-Ppp3ca and the recombinant adenovirus identified by enzyme digestion and PCR were in the right range corresponding with expectation. It was concluded that the recombinant adenovirus carrying rat calcineurin A alpha (Ppp3ca) cDNA as well as a report gene-enhancer green fluorescent protein gene was successfully constructed in this experiment.
Adenoviridae/*genetics ; Calcineurin/*biosynthesis ; Calcineurin/genetics ; Cloning, Molecular ; DNA, Complementary/genetics ; Genetic Vectors/genetics ; Green Fluorescent Proteins/biosynthesis ; Green Fluorescent Proteins/genetics ; Myocardial Reperfusion Injury/*genetics ; Myocardium/chemistry ; Rats, Wistar ; Recombination, Genetic/genetics

Objective: To discuss the way of early nutritional support in the patients after the liver transplantation. Methods: The nutritional support was used in 28 patients who had suffered the orthopetic liver transplantation (OLT).Parenteral nutrition was started as soon as possible after the operation. On the fourth day after the operation, enteral nutrition was also used. Results: 5 patients died from the postoperative complications. 23 patients survived and had been in better nutrition. Conclusion: The early nutrition support can play an important role in the patients recovery after OLT. EN should be administrated immediately when the gut functions.

To explore the relations of Morphology Immnuophe-notype Cytogenetics Molecular biology(MICM) detection on diagnosis andtreat,emt of leukemia． Methods: 68 cases of leukemia patients had beenanalyzed by morphology(FAB)． Immunohistochemistry(Flow Cytometry, FCM)． chromosome G banding technique and dual-color fluorescence insitu hybridization (D-FISH)．Technique:All patients were treated bychemotherapy． T test and X2 test of significance． Results: 7 cases have acute leukema aberration antigen expression． 5 out of 47 cases acutemyeliod leukemia patients accompany lymhocytic interrelated antigenexpression． 2/l5 cases acute lymphoid leukemia accompany myelocyteinterrelated antigen expression． 2 cases acute lmphoid leukemia are T cell and B cell interrelated antigen mingle expression． had been examined46,XY,t(8,2l) translocation of chromosome and bcr/abl fusion genes inthe acute leukemia patients． 16 out of 20 chronic myeloid leukemia patientshad philadelphia chromosome． 7 out of 20 patients had complicate karyotype． 5 out of 20 patients had bcr/abl fusion gene, l out of 4 patient had bcr/abl fusion gene that Ph chro mosome showed negative in CML． 3/4 cases patients had complicate chromosome． The ratio of CR use l time chemotherapy and the total ratio of CR using 2 times chemotherapywith aberration antigen expression in acute leukemia was significantly lessthan those of the acute leukemia patient had single system antigenexpression(P<0.05)． The time of CML-BC with complicate c hromosome karyotype was significantly short than those of Ph showed negative in CML(P<0.05)． Conclusion: The MICM classification is more acc urate for diagnosis of leukemia and more significant in guiding the leukemiatreamen．

Objective To observe the distribution features of phosphorylated p 44/42 extracellular signal-regulated kinase(p-ERK) induced by affective pain in the anterior cingulate cortex (ACC) of the rat.Methods Twelve male SD rats were randomly divided into the control group (n=6) and spinal nerve ligation (SNL) group (n=6).The rats of the SNL group were exposed to unilateral L5 spinal nerve ligation (SNL) surgery.The behavior was examined by mechanical pain thresholds (MPTs) and the pain affect was measured using open-field test, elevated zero maze.The expressions of p-ERK in ACC (AP 3.2,2.7,2.2mm from Bregma) were detected with immunofluorescence test .Results In comparison with the control group , the unilateral mechanical pain threshold in the SNL group decreased significantly after operation . The operation also resulted in pronounced anxiety-like behavior .The p-ERK labelled cells in different levels of ACC ( AP 3.2,2.7,2.2mm from Bregma) of SNL rats were (11.89 ±2.57),(32 ±4.67 ) and (17.56 ±2.04 ), respectively, while the control group rats were (12.44 ±2.16),(10 ±0.87 ) and (10.11 ±1.36).These data indicated that compared with the control group, the expressions of p-ERK in ACC(AP 2.7,2.2mm from Bregma) increased significantly in the SNL group after operation(P<0.01), besides for the level of ACC(AP 3.2mm from Bregma) (P>0.05).Conclusion The results suggest that the neuropathic pain can result in anxiety-like behavior and increased expressions of p-ERK in ACC of the rat.This change may be closely related to the expressions of p-ERK in ACC(AP 2.7,2.2mm from Bregma), but not to the level of ACC(AP 3.2mm from Bregma).