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.

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 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.

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

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 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 assess the awareness of nutrition issues among patients with type 2 diabetes mellitus (T2DM) and to find effective nutrition education models. Methods According to cluster random sampling method,256 T2DM patients from 2 hospitals in Zhengzhou City were enrolled and randomly assigned to the study or control group. The participants in the study group received traditional nutrition education and real object-based diabetic diet guidance, and those in the control group received traditional nutrition education only. Awareness about nutrition and blood glucose levels were compared between the two groups by using independent t test and Chi-square test Results After the intervention, the study group showed significantly higher nutrition scores ( 86. 5 ± 3.8 vs 71.5 ± 4. 6, P ＜ 0. 05 ) and lower 2 h postprandial glucose level ( ( 9. 15 ± 1.06 ) mmol/L vs ( 11.32 ± 0. 84) mmol/L, P ＜ 0. 05 ). Conclusion Real object-based diabetic diet teaching could foster the awareness in nutrition and decrease postprandial blood glucose level in T2DM patients.

Objective To explore the relationship between mechanical withdrawal threshold ( MWTs) and local cutaneous blood perfusion ( BP ) in rats with acute inflammation induced by carrageenan.Methods Twenty male Sprague-Dawley ( SD) rats were randomly divided into control and model groups.The acute inflammatory rat model was established by subcutaneously injecting with carrageenan into the left hindpaw.MWTs were measured by Dynamic Plantar Aesthesiometer 37450 before injection ( as base) of carrageenan and at 4 h, 24 h, 48 h, 72 h after carrageenan injection. The local cutaneous BP was detected by Pericam Perfusion Speckle Imager ( Pericam PSI) at the time after measuring of MWTs.The above two behaviors were compared and the relationship between them was analyzed.Results MWTs of the model group rats were decreased while BP significantly increased than that in the control group ( P<0.01 ) .The MWTs and BP in the model rats showed a negative correlation, especially the correlation index showed significant differences at 4 h and 72 h after carrageenan injection ( P<0.01) .Conclusions Carrageenan-induced acute inflammation in rats causes significant changes in mechanical pain threshold, which has a negative correlation with local cutaneous blood perfusion.

Objective To establish normal reference ranges for serum cystatin C (Cys C)in relatively healthy middle-aged and elderly(＞50 years)Chinese individuals.Methods A total of 1087 candidates were consecutively selected and serum Cys-C levels were measured by transmission turbidimetry.Frequency analysis and histogram were used to establish the 95％ confidence reference range according to methods provided by CLSI (C28-A2).Results Based on the definition and verification procedures for clinical laboratory reference ranges(CLSI C28-A2,second edition),Cys-C levels of 1087 participants fell within the range of 0.30-1.55 mg/L;Male participants had higher serum Cys-C levels than female participants(Z=-10.19,P＜0.01).The serum Cys-C level increased with age,regardless of gender(R =0.600,P＜ 0.01).Differences in Cys-C levels between age groups were statistically significant (x2=411.17,P＜ 0.01).The reference ranges of normal serum Cys-C levels for different age groups (50-,55-,60-,65-,70-,75-,) were 0.42-0.98mg/L,0.45-1.04mg/L,0.47-1.34mg/L,0.46-1.38mg/L,0.61-1.33mg/L,0.61-1.28 mg/L,respectively,for males,and 0.39-0.94mg/L,0.42-1.01mg/L,0.40-0.91mg/L,0.46-1.03mg/L,0.57-1.04mg/L,0.55-1.27mg/L,respectively,for females.Conclusions This study established preliminary normal serum Cys-C reference ranges for healthy middle-aged and elderly(＞ 50 years)individuals in this region,which can serve as parameters for disease diagnosis and treatment evaluation.