Objective To construct a sequence tag library of HepG2 cells by serial analysis of gene expression (SAGE). Methods Total RNA of HepG2 cells was extracted using a Trizol agent. The double strand cDNA were reverse-translated from mRNA which was isolated from total RNA. According to the protocol of SAGE, a sequence tag library of HepG2 cells was constructed. The transcript, which was represented by each tag, was analyzed by consulting the GenBank and UniGene, and the abundance of the tags was analyzed by SAGE analysis software. Results A total of 14 181 SAGE tags out of 15 586 tags were steadily and effectively amplified and the lengths of the tags were between 200-3000 bp. After sequence analysis of the tags, 2023 specific genes were revealed. Conclusions The use of improved reverse transcription agent kit may guarantee obtaining the full length cDNA, which provides materials for the construction of SAGE library. SAGE is a powerful method to investigate the gene expression profile, and it establishes a foundation for gene researches.

Objective To study the proliferation and differentiation patterns of hepatic stem cells from mice cultured in vitro isolated identify their characteristics.Methods Mice were divided into 5 groups according to the pregnant (embryo days,ED13,ED15 and ED18) or born age (day0 and 3).Hepatic stem cells were isolated and cultured in vitro.The amount of the stem cells as well as their growing situation and differentiation pattern were observed and compared among different groups and markers of stem cells (CD117,CD90.1,CD49f,c-Met),hepatic cells(AFP) were used to identify the cultures.Resulls The cells with best situation of growing as colonies were obtained from ED15 group.Their expression of specific markers suggested that they were hepatic stem cells.The stem cells isolated from ED15 mice in subculturing proliferated in line pattern and differentiated in reverse line pattern.The expression of AFP varied as normal distribution as cell differentiation development.Conclusion Most cells have characteristics of hepatic stem cells isolated from fetal liver and the number of these cells decreases gradually as embryo duration prolongs.The hepatic stem cells proliferate in line pattern and differentiate into hepatic cells after in vitro culture.

Objective To observe the availability and safety of ventricular septal defect (VSD) occluder in infants and young children with large patent ductus arteriosus (PDA) associated with severe pulmonary hypertension.Methods Five patients (1 male and 4 fomale) of large PDA aged 5 months to 3 years,weighted from 5.1 to 15 kg,body surface area (BSA) 0.37-0.58 m2 underwent transcathter intervention with concentric VSD occluders from June 2008 to May 2009.Arterial ducta were tube-like and their diameters were 5.7 to 8.5 mm,with ulmonary vascular resistance from 4.8 to 5.7 Wood Unit,Qp/Qs 3.4-4.6.Three patients were given Bosentan after intervention.Results The large PDAs were successfully closed with VSD occluders,including 1 concentric perimembranous VSD occluder and 4 muscular VSD occluders.They all discharged 4 to 5 days with hidrosis and weight improved.Echocardiogram indicated VSD occluder was stable,no residue shunt and no stricture of left pulmonary artery and descending aorta were found.According to tricuspid and pulmonary regurgitation,pulmonary arterial pressure decreased differently and returned to normal after 6 months follow-up.Conclusion VSD occluder is available and effective to close large PDA associated with severe pulmonary hypertension in inrants and young children,but more cases and long-term follow-up are necessary.

Objective To develop an ideal cultural method to amplify embryonic hepatic stem cells and inhibit their differentiation in vitro. Methods Suspension of ED 14 Fischer (F) 344 rat em-bryonic hepatic stem cells was prepared by collagenase digestion and mechanical disaggregation. Then cells were divided into two groups randomly. The cells in group 1 were seeded into type I collagen-coated plates by adherent culture while those in group 2 were seeded into soft agar medium by suspen-sion culture. After culture for 2 weeks, the morphology and ultrastructure of cells in both groups were observed and compared by inverted microscope and transmission electron microscope, respectivley.The expression of CD90. 1 and CD49F, the two specific stem cell surface markers, was tested by flow cytometry to manifest the establishment of embryonic hepatic stem cells. Alkaline phosphatase stai-ning was used to detect stem cell differentiation. Result Embryonic hepatic stem cells in group 2 were characterized by higher nucleus-cytoplasm ratio and less cell organelles, higher expression of CD90. 1 and CD49F, and stronger positive reaction for alkaline phosphatase staining compared with those in group 1. Moreover, the cells in group 1 showed significant differentiation features. Conclusion Em-bryonic hepatic stem cells cultured suspendedly in soft agar medium experience less differentiation than those adherently cultured in serum-added culture medium, and can proliferate and form clone ball with a specific stem cell feature.

Aim To investigate the effect of DHA on NGF-induced neuronal differentiation of PC12 cells and explore the possible mechanism via regulating BMP pathway. Methods PC12 cells were treated with 100μg·L-1 NGF and 100 μg·L-1 NGF + 10 μmol· L-1 DHA for 3, 6 and 9 days respectively. The length and number of neurite were detected by immunofluores-cenc. DHA content was analyzed by gas chromatogra-phy in all groups. The protein expression of BMP4, BMP7 , BMPR-II and p-Smad 1/5/8 was determined by Western blot. Results The length of total primary neurite in NGF+DHA groups was obviously increased, longer than that in NGF group; DHA content in 10μmol · L-1 DHA group was higher than that in the control group;NGF+DHA groups also unregulated the protein expression of BMP4 , BMP7 , BMPR-II and p-Smad 1/5/8 . Conclusion DHA promotes NGF-in-duced neuronal differentiation in PC12 cells, which may be associated with the upregulation of BMP path-way protein.

Objective To observe the protective effect of pretreatment with wortmannin against pancreas and liver injuries indued by severe acute pancreatitis(SAP) in rats and investigate its mechanism.Methods Fifty-four SD rats were randomly divided into 3 groups: control group(C group),SAP group(P group) and SAP+wortmannin group(PW group)(n=18 per group). SAP model was induced by retrograde infusion of 50g/L sodium taurocholate into the biliopancreatic duct of rats,except C group in which sodium taurocholate was replaced by normal saline. Serum level of tumor necrosis factor-alpha(TNF-?)、 alanine aminotransferase(ALT)、aspartate aminotransferase(AST)、and nuclear factor-kappa B(NF-?B)in liver were detected. Histopathology of liver and pancreas was studied.Results In P group, serum levels of TNF-?、ALT、AST and NF-?B in liver were significantly elevated(P

Objective This study was to evaluate the usefulness of the model of end-stage liver disease (MELD) score in comparison with the Child-Pugh classification to predict postoperative mortality and short-term survival in liver transplant patients. MethodsMELD score and Child-Pugh score were computed for each patient according to the original formula on admission day. Kaplan-Meier survival curves were made using the cut-off points calculated by Youden index. ResultsSix out of 40 patients died within three months, MELD scores and Child-Pugh scores for non-survivors (32.2?8.0, 12.3?2.0) were higher than those for survivors (13.4?6, 9.12?2.31) significantly (P

To investigate the effect of three kinds of polymeric scaffolds on attachment, proliferation and differentiation of bone marrow mesenchymal stem cells, the cells were different polymeric scaffolds of PLA-PEG, PLA, PLGA, respectively. The proliferation of cell was evaluated by cell count; the attachment and morphology of BMSCs were observed by SEM; and differentiation was detected by alkaline phosphatase activity, fluorescence, and RT-PCR methods. Results showed that the cells in PLGA group spread better among BMSCs adhered to the three polymeric scaffolds. The activity of ALP was detected after 3 days culture in these three groups. There were no significant differences between PLA-PEG and PLGA groups, but the activity of ALP was higher than PLA group. The gene expressions of osteocalicin and collagen I were also observed in the early culture time. Calcium nodes formation in these polymeric scaffolds were detected. BMSC spreading first, then overlapping growth and secretion of matrix around the bottom and surface of scaffolds were observed through SEM. In summary, PLA-PEG and PLGA are better polymeric scaffolds for the bone tissue engineering, compared with PLA.
Bone Marrow Cells ; cytology ; Cell Adhesion ; Cell Proliferation ; Lactic Acid ; chemistry ; Mesenchymal Stromal Cells ; cytology ; Polyesters ; chemistry ; Polyethylene Glycols ; chemistry ; Polyglycolic Acid ; chemistry ; Polymers ; chemistry ; Tissue Engineering ; Tissue Scaffolds

A novel bioinspired phospholipid copolymer has been synthesized by the radical polymerization of poly2-Methacryloyloxyethylphosphorylcholine (MPC), stearyl methacrylate (SMA), hydroxypropyl methacrylate (HPMA) and trimethoxysilylpropyl methacrylate (TSMA). Contact angle results indicated that the coating surface rearranged to get a more hydrophilic surface at the polymer/water interface. The membrane mimic phosphorylcholine coating surface could resist the platelet adhesion and prolong plasma recalcification time significantly. Rapamycin was used as model drugs to prepare drug-eluting coating. The animal experiments showed that this novel drug-eluting stent could effectively prevent the phenomena of restenosis.
Angioplasty, Balloon, Coronary ; instrumentation ; Animals ; Coated Materials, Biocompatible ; Coronary Restenosis ; prevention & control ; Drug-Eluting Stents ; Female ; Humans ; Male ; Materials Testing ; Methacrylates ; chemistry ; Phosphorylcholine ; analogs & derivatives ; chemistry ; Pilot Projects ; Polymers ; chemistry ; Prosthesis Design ; Random Allocation ; Sirolimus ; chemistry ; Swine ; Swine, Miniature

Objective To investigate the expression levels of serum miR-210and miR-375in patients with non-small cell lung cancer (NSCLC).Methods A total of 25NSCLC patients (NSCLC group) and 14healthy volunteers (control group) were enrolled in this study.The relative expression levels of 9miRNAs (miR-182, miR-126, miR-31, miR-21, miR-221, miR-200b, miR-183, miR-210and miR-375) in 6 NSCLC patients and 6healthy volunteers were measured by RT-qPCR.The dysregulated miRNAs will be selected as candidate miR-NAs.The diagnostic value were evaluated by ROC curve.Results Compared with control group, 2 (miR-210and miR-375) out of 9miRNAs were up-regulated in NSCLC group, and the differences were statistically significant (P＜0.05), while the other 7miRNAs were not consistent with the reported literatures.Therefore, miR-210and miR-375were selected as candidate miRNAs.We found that the relative expression level of miR-210in the lung adenocarcinoma group was significantly different from control group (P＜0.05), while there was no significant difference between the squamous cell carcinoma group and the control group (P＞0.05).There was no significantly statistical difference in the relative expression level of miR-375between lung squamous cell carcinoma group, lung adenocarcinoma group and the control group (P＞0.05).The AUC of serum miR-210of lung adenocarcinoma group was 0.737 5 (95％CI:0.498 3-0.976 7, P=0.091 4) with a medium diagnostic value.Conclusion MiR-210is highly expressed in the serum of patients with lung adenocarcinoma, suggesting that miR-210may be a novel tumor marker for the diagnosis of lung adenocarcinoma.The value of miR-375in the diagnosis of lung cancer still needs to be further explored.