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.

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 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 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.

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) .

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

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 explore the effects of SLC concentration gradient on suppression of tumor immune escape. Methods According to different SLC concentration, there were six groups. The HLA-Ⅰexpression and apoptosis of MCF-7 were detected with FCM,and intracellular BCL-2 expression was analyzed by western blot. The production of TGF-? was detected with ELISA. Results In a certain range of concentration gradient, following SLC increase, HLA-Ⅰexpression level on MCF-7 was improved, and apoptosis was induced but BCL-2 expression was enhanced. Moreover, the secretion of TGF-? was suppressed. Conclusion SLC inhibites tumor immune escape.

Objective:To explore whether Chinese herbs Astragalus and Angelica mixture(A&A) have renoprotective effects on chronic renal failure in rats.Methods:5/6 nephrectomized kidney in rats was used as chronic renal failure model.Rats were randomly divided into four groups which included sham,Model,A&A and enalapril(Ena) groups.The remnant kidney was harvested after 2,4,8,12 weeks,and levels of serum creatinine(scr),urea and urinary protein excretion were measured using routine biochemical methods.The pathological lesions were observed by light microscope.The expressions of fibronectin(FN) and collagen Ⅳ(Col-Ⅳ) in glomeruli and interstitium were detected by immnuohistochemistry.Results:5/6 nephrectomized rats were presented with the characteristics of increasing scr and urea,persistent proteinuria at the end of the 2nd,4th,8th,12th weeks,respectively.They had increasing size of glomeruli with progressive proliferation of mesangial cells and mesangial matrix followed by glomerulosclerosis and progressive elevated expressions of FN and Col-Ⅳ.Compared with Model group,the administration of A&A could decrease urinary protein excretion for 34%,44%,25%,45% at the end of the 2nd,4th,8th and 12th weeks,respectively(P

12 kinds of histochemical methods were adopted to evaluate the effects of gossypol optical isomers on the reproductive system of Wistar rats, following the administration of (?) and (+)-gossypol at a dosage of 30mg/kg daily, and(-)-gossypol at a dosage of 15 mg/kg daily, respectively for 2 or 3 weeks. (+)-gossypol has no an tifertility activity. (?) and (-)-gossypol caused kinetic disorder of spermatogemsis, and inhibited turnover of nuclear basic protein of the late, elongated spermatids in the testis. No changes in G6PDH and 30-HSDH histochmical reaction in Leydig cells, and ALP in the basement membrane of epididymis were observable reflecting that gossypol has no effect on synthesis and secretion of testosterone. Spermatozoa in the epididymis of rats treated with (?) and (-)-gossypol exhibited head and tail separation. The formazan deposits of reaction of LDH, LDH-X, MDH and ICDH located in sperm mitochondria appeared to become coarse and to reduce in quantity after (?) and (-)-gossypol treatment.