Search

Objective: To clone, express and purify Schistosoma japonicum fructose-1, 6-bisphosphate aldolase (SjFBPA) in E. coli and observe its expression in different developmental stages of S. japonicum., Methods: FBPA gene was amplified from S. japonicum adult worm cDNA by using PCR. The amplified product was recombined into pET28a plasmid, and inducibly expressed with IPTG in E. coli BL21. SDS-PAGE and Western blotting were employed to analyze and identify the recombinant protein SjFBPA (rSjFBPA). Then, rSjFBPA was purified by chromatographic purification and its purity was analyzed by SDS- PAGE. The protein concentration of rSjFBPA purified was measured by the BCA method. Furthermore, SjFBPA mRNA was ana- lyzed in different developmental stages of S. japonicum by RT-PCR., Results: SjFBPA was successfully amplified by using PCR and identified by restriction enzyme digestion and sequencing. The Western blotting analysis confirmed that the recombinant pro- tein could specifically reactive to the anti-His-tag monoclonal antibody. The concentration of the purified recombinant protein was about 4 mg/ml. The result of RT-PCR showed that SjFBPA mRNA was expressed in cercaria, schistosomulum, adult worm and egg of S. japonicum., Conclusion: SjFBPA is successfully recombined and expressed in a prokaryotic system, and SjFBPA mRNA is expressed in cercaria, schistosomulum, adult worm and egg of S. japonicum.

The excessive apoptosis of cells of the nucleus pulposus may plays an important role in intervertebral disc (IVD) degeneration. It has been shown that the pro-inflammatory cytokine tumour necrosis factor (TNF)-α can induce disc cell apoptosis. Insulin-like growth factor (IGF)-1 can promote nucleus pulposus cell proliferation; however, whether or not IGF-1 inhibits TNF-α-induced apoptosis in the nucleus pulposus has not yet been elucidated. In this study, our objective was to create a potentially therapeutic viral vector, which could be used to achieve the enforced expression of IGF-1 in rabbit nucleus pulposus cells. Furthermore, we investigated the ability of IGF-1 to reverse TNF-α-induced apoptosis in cells of the nucleus pulposus. Isolated nucleus pulposus cells were cultured to a confluent monolayer, digested with collagenase Ⅱ and purified using trypsin and differential adhesion methods. Nucleus pulposus cells were positively identified using type Ⅱ collagen immunohistochemistry. Following transfection with adenoviral vectors engineered to overexpress recombinant human IGF-1 (Ad-hIGF-1) or TNF-α, the cells were observed under a light microscope. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end-labeling (TUNEL) and flow cytometry (FCM) were used to assess the rate of apoptosis. The Ad-hIGF-1 viral vector was effectively transduced into the nucleus pulposus cells and increased IGF-1 expression as confirmed by RT-PCR and western blot analysis. In the TNF-α-treated group, a large number of apoptotic cells was observed that exhibited morphological changes associated with this form of cell death. Minimal apoptosis was observed in the Ad-hIGF-1-treated group and the control group showed no obvious signs of apoptosis. TUNEL assay revealed that the rate of apoptosis in the Ad-hIGF-1 group was significantly reduced compared with the TNF-α-treated group (P<0.01). This result was confirmed using FCM. The rate of apoptosis was also significantly increased in the TNF-α-exposed cells compared with the control group (P<0.01). Our findings strongly suggest that the adenoviral vector expressing hIGF-1 can successfully infect nucleus pulposus cells in vitro and effectively enhance the expression of IGF-1. In addition, IGF-1 reversed the TNF-α -induced apoptosis of nucleus pulposus cells. Thus, Ad-hIGF-1 may be useful in the development of clinical interventions for disc degeneration.

The conditioned medium from B104 neuroblastoma cells (B104CM) induces proliferation of oligodendrocyte progenitor cells (OPCs) in vitro. Our previous study showed that phosphorylation of extracellular signal-regulated protein kinase(s), not PI3K or p38, is key to B104CM-induced OPC proliferation. However, whether there are still other signaling pathways that are also involved in B104CM-induced proliferation remains unknown. In the present study, we evaluated the implication of c-Jun N-terminal kinase (JNK) signaling pathway in the B104CM-induced proliferation of OPCs using the specific inhibitor of JNK. We provided convincing evidence for the first time that the phosphorylation of JNK is necessary for OPC proliferation induced by B104CM in vitro. Moreover, the activation of JNK results in subsequent expressions of c-jun, c-fos, and c-myc, which initiates proliferation of OPCs. Collectively, these results suggest that JNK is also necessary for B104CM-induced OPC proliferation.

We present a case of a 58-year-old female with a rare vascular tumor of intermediate malignancy. The initial manifestation was a pseudoaneurysm caused by the rupture of the right pulmonary artery after tumor invasion. The diagnosis of epithelioid hemangioendothelioma was confirmed by the morphologic and immunocytochemical features after surgery. The patient recovered smoothly and there has been no evidence of local recurrence or metastasis during the 2 years of follow-up., (Georg Thieme Verlag KG Stuttgart · New York.)

Objective: To explore the effect of land consolidation project on the detention of Oncomelania hupensis snails in the flood at the Yangtze River beach., Methods: The land consolidation project including removing wild willow groves, cutting the high and filling the low, and improving the slope was carried out at the Yangtze River beach in 2011, and the status of snails was investigated before and after the project, respectively, at the end of the flood season., Results: Before the project (2010), at the end of the flood season, the rate of the frames with snails was 96.5% and the snail density was 3.655/0.1 m2. After the project (2012), at the end of the flood season in the same area, the rate of the frames with snails was 2.0% and the snail density was 0.015/0.1 m2 (chi2 = 357.29, P < 0.01)., Conclusion: The land consolidation has a significant effect on the detention of snails in the flood at the Yangtze River beach.

This study aims to find good markers for predicting the prognosis of patients with esophageal squamous cell carcinoma (ESCC). Vasculogenic mimicry (VM) and the expression of hypoxia inducible factor-lα (HIF-lα)/E-cad protein in ESCC were investigated by immunostaining. The association between VM, HIF-lα/E-cad and clinicopathologic characteristics and 5-year-survival rate of patients with ESCC was analyzed. A total of 160 ESCC specimens were involved in this study and 28 specimens of normal esophageal mucosa served as controls. VM channels were identified in 78 (48.75%) of the 160 ESCC specimens and none of the normal esophageal mucosa was found to have VM. The rates of high-expression of HIF-lα and E-cad in ESCC were 43.75% and 38.75%, while the rates in control were 17.86% and 71.43%, respectively (P<0.05 for all). VM and the expression levels of HIF-lα and E-cad were significantly related to lymph node metastasis, serosa infiltration, PTNM staging and 5-year-survival rates of patients with ESCC (P<0.05 for all). VM was positively correlated with HIF-lα but negatively with E-cad, and HIF-lα was negatively correlated with E-cad (P<0.001 for all). The 5-year-survival rate of patients with ESCC was 6.41% (5/78) in VM group and 65% (52/82) in non-VM group, 7.14% (5/70) in high HIF-lα expression group and 57.78% (52/90) in low HIF-lα expression group. Oppositely, the 5-year-survival rate in high E-cad expression group was 80.65% (50/62) and that in low E-cad expression group was 7.37% (7/98) (P<0.05 for all). Cox multifactor regression analysis indicated that lymph node metastasis, PTNM stage, VM and expression levels of HIF-lα and E-cad were independent risk factors of patients with ESCC (P<0.05 for all). Combined detection of VM, HIF-lα and E-cad plays an important role in predicting the invasion, metastasis and prognosis of patients with ESCC.

The conditioned medium from B104 neuroblastoma cells (B104CM) induces proliferation of oligodendrocyte progenitor cells (OPCs) in vitro. However, the molecular events that occur during B104CM-induced proliferation of OPCs has not been well clarified. In the present study, using OPCs immunopanned from embryonic day 14 Sprague-Dawley rat spinal cords, we explored the activation of several signaling pathways and the expression of several important immediate early genes (IEGs) and cyclins in OPCs in response to B104CM. We found that B104CM can induce OPC proliferation through the activation of the extracellular signal-regulated kinases 1 and 2 (Erk1/2), but not PI3K or p38 MAPK signaling pathways in vitro. The IEGs involved in B104CM-induced OPC proliferation include c-fos, c-jun and Id2, but not c-myc, fyn, or p21. The cyclins D1, D2 and E are also involved in B104CM-stimulated proliferation of OPCs. The activation of Erk results in subsequent expression of IEGs (such as c-fos, c-jun and Id-2) and cyclins (including cyclin D1, D2 and E), which play key roles in cell cycle initiation and OPC proliferation. Collectively, these results suggest that the phosphorylation of Erk1/2 is an important molecular event during OPC proliferation induced by B104CM.

Excessive apoptosis in intervertebral disc (IVD) cells is important in IVD degeneration. Interleukin (IL)-1β has been shown to induce apoptosis in these cells. However, whether insulin-like growth factor-1 (IGF-1) inhibits IL-1β-induced apoptosis in the nucleus pulposus remains unclear. The purpose of this study was to investigate the effects of IGF-1 on IL-1β-induced apoptosis in the nucleus pulposus. Cells isolated from the nucleus pulposus were grown in culture to a monolayer. These cells were identified using immuno-histochemistry for type II collagen and toluidine blue staining for glycosaminoglycans. Following exposure to IGF-1 or IL-1β, the cells were observed using light microscopy. Giemsa staining, TdT-mediated dUTP-biotin nick end-labeling (TUNEL) and flow cytometry (FCM) were used to detect the rate of early cell death, which served as an indicator of apoptosis. In the IL-1β group, a large number of these cells underwent apoptosis and demonstrated morphological changes associated with apoptosis. A small proportion of cells exposed to IGF-1 alone underwent apoptosis. No obvious signs of apoptosis were observed in the control group. TUNEL results revealed that the rate of apoptosis in the IGF-1 group was significantly reduced compared with that in the IL-1β group (P<0.01), confirmed using FCM. Compared with the control group, the apoptotic rate was also significantly increased in IL-1β-exposed cells (P<0.01). These findings strongly suggested that IGF-1 inhibits IL-1β-induced apoptosis in the nucleus pulposus.

Neural stem cell (NSC) transplantation is a major focus of current research for treatment of spinal cord injury (SCI). However, it is very important to promote the survival and differentiation of NSCs into myelinating oligodendrocytes (OLs). In this study, myelin basic protein-activated T (MBP-T) cells were passively immunized to improve the SCI microenvironment. Olig2-overexpressing NSCs were infected with a lentivirus carrying the enhanced green fluorescent protein (GFP) reporter gene to generate Olig2-GFP-NSCs that were transplanted into the injured site to differentiate into OLs. Transferred MBP-T cells infiltrated the injured spinal cord, produced neurotrophic factors, and induced the differentiation of resident microglia and/or infiltrating blood monocytes into an "alternatively activated" anti-inflammatory macrophage phenotype by producing interleukin-13. As a result, the survival of transplanted NSCs increased fivefold in MBP-T cell-transferred rats compared with that of the vehicle-treated control. In addition, the differentiation of MBP-positive OLs increased 12-fold in Olig2-GFP-NSC-transplanted rats compared with that of GFP-NSC-transplanted controls. In the MBP-T cell and Olig2-GFP-NSC combined group, the number of OL-remyelinated axons significantly increased compared with those of all other groups. However, a significant decrease in spinal cord lesion volume and an increase in spared myelin and behavioral recovery were observed in Olig2-NSC- and NSC-transplanted MBP-T cell groups. Collectively, these results suggest that MBP-T cell adoptive immunotherapy combined with NSC transplantation has a synergistic effect on histological and behavioral improvement after traumatic SCI. Although Olig2 overexpression enhances OL differentiation and myelination, the effect on functional recovery may be surpassed by MBP-T cells.

The conditioned medium from B104 neuroblastoma cells (B104CM) induces neural stem cells (NSCs) to differentiate into OPCs in vitro, which indicates that certain factor(s) contained within the B104CM must give instructional signals that direct OPC differentiation of NSCs. However, the OPC-inductive factor(s) present within the B104CM has not been well identified yet. Platelet-derived growth factor AA (PDGF-AA) was not only known to be a potent mitogen for OPC proliferation but also to act as a regulator of oligodendrocyte differentiation from multipotent embryonic NSCs. This raises the possibility that B104CM induces OPC differentiation of NSCs through secretion of PDGF-AA. In the present study, we detected the expression of PDGF-AA mRNA in B104 cells and the high level of PDGF-AA protein in B104CM. Most importantly, B104CM-induced OPC differentiation of NSCs could be completely blocked by AG1295, a specific inhibitor of PDGFR signal pathway, suggesting that the PDGF-AA in B104CM is the key factor that induces NSCs to differentiate into OPCs. Moreover, such B104CM-induced OPC differentiation appears to be mediated by the extracellular signal-regulated kinases 1 and 2 (Erk1/2), phosphatidylinositol-3 kinase (PI3K), and p38 signal pathway because B104CM elicited the activation of Erk1/2, PI3K, and p38, which could be markedly blocked by U0126, LY294002, and SB203580, several specific inhibitors of these signal pathway, respectively. These inhibitors also abolished OPC differentiation of NSCs completely. Together our study suggests that PDGF-AA contained in B104CM is the key regulating molecule that instructs OPC differentiation from embryonic NSCs through the activation of Erk, PI3K, and p38 signal pathway in vitro.

Bone morphogenetic proteins (BMPs), a subgroup of the TGF-β superfamily, play critical roles in neural progenitor cell fate determination. Neural stem cells (NSCs) are multipotent progenitor cells that can differentiate into neurons, oligodendrocytes and astrocytes under certain conditions. In our recent report, using an antibody that can recognize both BMP-2 and BMP-4 (BMP-2/4), we showed that BMP-2/4 is only expressed in astrocytes differentiated from NSCs in a medium containing 1% fetal bovine serum (FBS). In this in vitro model, the astrocytic differentiation of NSCs was mainly toward type-2. When NSCs were cultured in a medium containing 10% FBS, most of the cells differentiated into type-1 astrocytes. However, little information is available for BMP-2 and BMP-4 expression in type-1 and type-2 astrocytes induced from NSCs under these different culture conditions. In this study, using two antibodies specific for BMP-2 and BMP-4, respectively, we discriminated the presence of BMP-2 and BMP-4 in NSCs and their derivatives under 1% and 10% FBS culture conditions by RT-PCR, western blot and immunofluorescence staining. We found that BMP-2 and BMP-4 are highly expressed in both type-1 and type-2 astrocytes, and no detectable expression in NSCs, neurons and oligodendrocytes. This suggests that the astrocytes might be one source of BMPs during the differentiation of NSCs. However, in our model, we cannot exclude the possibility that microglia or endothelial cells could also be a source of BMPs.

Olig1, a member of class B basic-helix-loop-helix (bHLH), plays key roles in early oligodendrocyte specification. Inhibitors of DNA binding (Id) is another sub-class of HLH proteins, act as dominant-negative regulators of bHLH proteins, which can form heterodimers with class A or B bHLH proteins, but lack the critical basic DNA binding domain. Id4 was recently found to interact with olig1 and inhibit oligodendrocyte differentiation. However, there still no direct evidence to reveal the spatial and temporal interaction of olig1 and ID4 in living cells. In this study, we performed bimolecular fluorescence complementation (BiFC) analysis to further characterize the distinct subcellular localization of olig1, ID4 and their dimer in living SW1116 cells. To examine the subcellular localization of olig1 and ID4 by themselves, the olig1-EGFP or ID4-DsRed2 fusion proteins were also expressed in SW1116 cells, respectively. As predicted, the olig1-EGFP fusion proteins were located in the nucleus, and ID4-DsRed2 fusion proteins were located in the cytoplasm. When olig1-EGFP and ID4-DsRed2 fusion proteins were co-expressed, the green and red signals were co-located in the cytoplasm. Using BiFC, the strong BiFC signals could be detected in pBiFC-olig1VN173 and pBiFC-ID4VC155 co-transfected cells and the fluorescence signal was located in the cytoplasm. These results collectively confirmed that olig1 and ID4 could interact and form dimer in living cells, and ID4 could block the transport of olig1 from cytoplasm to nucleus.

The PKC signaling pathway has been implicated in diverse cellular functions. Here, we sought to investigate the role of PKC-alpha÷beta in C6 glioma cell migration. We found that both PKC-alpha and PKC-beta were expressed by C6 glioma cells, but only PKC-alpha was markedly activated in serum-treated C6 cells. Go6976, a PKC-alpha÷beta specific inhibitor, was found to cause a dose-dependent reduction of PKC-alpha activation and cell migration induced by serum in C6 cells. These results collectively indicated that the PKC-alpha signaling pathway is necessary for glioma cell migration. Our findings may provide an insight into a better understanding to the malignant progression of gliomas.

Objective: To examine the number and function of circulating endothelial progenitor cells (EPCs) in children with cyanotic congenital heart diseases (CHD) and study their correlation with serum levels of vascular endothelial growth factor (VEGF) and stromal cell derived factor-1 (SDF-1)., Methods: Fifteen children with tetralogy of Fallot (cyanotic group) and 15 age-and sex-matched children with ventricular septal defect (control group) were enrolled. Serum levels of VEGF and SDF-1 were measured using ELISA. Mononuclear cells were isolated from peripheral blood by Ficoll density gradient centrifugation and cultured in vitro. EPCs were identified by immunofluorescence and were counted under a microscope. Modified Boyden chamber assay and the MTT assay were used to measure the migration and proliferation capacities of EPCs. EPCs adhesion ability assay was performed by replating cells on fibronectin-coated dishes, and then adherent cells were counted. The correlations of serum levels of VEGF and SDF-1 with the number and function of circulating EPCs were assessed by linear regression analysis., Results: Serum levels of VEGF (201.42+/-44.74 ng/L vs 113.56+/-35.62 ng/L; P<0.05) and SDF-1 (3.45+/-1.07 ng/L vs 1.05+/-0.99 ng/L; P<0.05) in the cyanotic group were higher than those in the control group. There was a positive correlation between serum levels of VEGF and SDF-1(r=0.675, P<0.01). The number of EPCs (*200 field) in the cyanotic group significantly increased compared with that of the control group (72.2+/-9.73 vs 51.2+/-3.83; P<0.01). The functional activities of EPCs, including proliferation, migration and adhesion capacities, were augmented in the cyanotic group compared with those in the control group. The increased number and function of EPCs and the increased serum levels of VEGF and SDF-1 were consistent in the cyanotic group, with a correlation coefficient of 0.8395, 0.5491, 0.6376 and 0.7392 respectively., Conclusions: The number and functional activity of EPCs as well as serum levels of VEGF and SDF-1 increased in children with cyanotic CHD. Serum levels of VEGF and SDF-1 were correlated to the number and functional activity of EPCs. Serum VEGF and SDF-1 together with circulating EPCs may play important roles in the pathology and physiology in these patients.

Objective: To analyze the differential expression proteins of rat ischemia/reperfusion (I/R) lung tissues in vivo and normal lung tissues by comparative proteome analysis, and to study the mechanism of donor lung I/R injury., Methods: Forty male SD rats were randomly divided into 2 equal groups: I/R group undergoing mimic orthotopic left lung auto-grafting and harvesting of the left lung five hours after the operation, and control group undergoing isolation of the left hilus of lung and then harvesting of the left lung. The differential proteins in the left ventricle of transplanted heart were separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE), identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and searched through Matrix Science software system. Western blotting was used to verify part of the differentially expressed proteins., Results: Well-resolved and reproducible 2-DE profile of rat I/R lung tissues and normal lung tissues were obtained. In the I/R lung tissue profile, the average spot number from 3 gels was 489 +/- 52 spots (P > 0.05) with an average matching rate of 89.28% (P > 0.05), and in the control group, the average spot number from 3 gels was 511 +/- 83 spots (P > 0.05) with an average matching rate of 91.22% (P > 0.05). Fourteen differential proteins were identified by peptide mass fingerprinting (PMF) searched in Matrix Science (P < 0.05). Western blotting confirmed that the protein expression of selenium binding protein 1 (SBP-1) and heat shock protein 25 (HSP25) increased at the early stage of I/R group., Conclusion: The protein expression of HSP25 and SBP-1 with stress protection function is upregulated in the early stage of lung I/R injury. Other differentially expressed proteins identified may have important functions in energy metabolism, tissue stress, cell apoptosis, and signal transduction.

Objective: To summarize the clinical application of bioprosthetic valve replacement., Methods: Fifty two patients, aged 13-73(52.4+/-14.0) years, underwent cardiac valve replacement with biovalves from June 2002 to June 2006 in our hospital. Fifty three tissue valve replacements combined with 8 mechanical valve replacements were performed. Other procedures were also carried out if appropriate, including bidirectional Glenn shunt in 1 patient with double outlet of right ventricle and pulmonary artery stenosis; interruption of accessory pathway in 1 patient with Wolff-Parkinson-White syndrome; coronary artery bypass graft in 5 with severe coronary artery disease; and atrial or ventricular repair in 4 with congenital septum defects., Results: One patient died in hospital due to multiple organ failure. The hospital mortality rate was 1.9%. The mean follow-up was 25 months. Two patients lost follow-up and the follow-up rate was 96%. Forty one patients returned to NYHA class I, 9 to class II, and 1 to class III. Echocardiography showed the implanted bioprosthesis functioned well without stenosis or regurgitation. There was no structural deterioration with freedom from thromboembolism and anticoagulant hemorrhage in all patients., Conclusion: The efficacy of biovalves in heart valve replacement is satisfactory.

Objective: To review the indications and effect of the intra-aortic balloon pump (IABP) in 38 patients during the cardiac surgery., Methods: From June 2004 to August 2007, 38 patients were performed IABP in the cardiac surgery, including 28 coronary artery bypass grafting (CABG) and 10 valve replacements. The intra-operation prophylactic use of IABP was in 5 cases, the use of IABP for weaning from cardiopulmonary bypass (CPB) during the cardiac surgery was in 22 cases, and the rest use of IABP was after the operation in the intensive care unit. Low molecular weight heparin was used for IABP anticoagulation., Results: Thirteen died out of the 38 IABP patients. The total IABP success rate was 66%.The IABP success rate in CABG was 82%, and 20% in the valve replacement. The patients with intra-operation prophylactic use of IABP all recovered, and 16 recovered out of the 22 patients with IABP uses for weaning from CPB, and 6 recovered out of the 11 patients with post-operation IABP application. No severe complications related to IABP occurred., Conclusion: IABP has better effect in CABG patients than in valve replacements. Effect of the prophylactic use of IABP is better than both the IABP use for weaning from CPB or the post-operation IABP application. For IABP anticoagulation, low molecular weight heparin is the better alternative than routine heparin, and the proper ACT is controlled at 150-170 s.

Objective: To summarize the experience of clinical diagnosis and surgical treatment of ascending aortic aneurysm., Methods: The clinical data of 72 cases of ascending aortic aneurysm were retrospectively analyzed. Of the 72 cases, 36 suffered Mafanos syndrome, 35 suffered aortic dissection, and 1 multiple aortic pseudo-aneurysm. All the patients received artificial vessel replacement or repair under middle or low temperature of cardiopulmonary bypass. Two patients received coronary artery bypass grafting, 2 received aortic valve replacement,and 1 received artificial patch repair., Results: Eight patients died and the others recovered without nervous system complications and were discharged., Conclusion: Expedient confirmatory diagnosis, prompt surgical therapy, and efficient therapy during the peri-operative period are helpful to achieve a successful outcome.

A microwave digestion-flame atomic absorption spectroscopy (FAAS) method was developed for the determination of metal elements Na, Zn, Cu, Fe, Mn, Ca and Mg in Mongolian patents. The instrument parameters for the determination were optimized, and the appropriate digestion solvent was selected. The recovery of the method was between 95.8% and 104.3%, and the RSD was between 1.6% and 4.2%. The accuracy and precision of the method was tested by comparing the values obtained from the determination of the standard sample, bush twigs and leaves (GSV-1) by this method with the reference values of GSV-1. The determination results were found to be basically consistent with the reference values. The microwave digestion technique was applied to process the samples, and the experimental results showed that compared to the traditional wet method, the present method has the merits of simplicity, saving agents, rapidness, and non-polluting. The method was accurate and reliable, and could be used to determine the contents of seven kinds of metal elements in mongolian patents.

Objective: To evaluate the therapeutic effect of remaining integrated mediastinal pleura upon the aortic arch and performing the anastomosis at the left cervix in radical operation for esophageal carcinoma., Methods: Ninety-eight patients with esophageal carcinoma were treated with the operation mentioned above. Among them, 56 patients had cancer in the middle, 12 in the upper-middle, 24 in the lower-middle segments, and 6 had double-primary tumors, with carcinoma length of (5.2+/- 2.4) cm. The TNM stages were 6 of Stage I and 92 of Stages II-III. All cases were squamous cell carcinomas., Results: All patients had satisfactory operation processes, without perioperative death, chylothorax, dyspnea, gastric retention, incision infection, and severe gastro-esophageal reflux. The life quality of the patients was improved., Conclusion: The radical operation with remaining integrated mediastimal pleura upon the aortic arch and anastomosis at the cervix for treating esophageal carcinomas is of minimal invasion with fewer complications, and may be used in clinical practice.

Objective: To investigate the effect of decellular treatment on the framework constituents of extracellular matrix and tissue stability in bovine jugular vein conduit (BJVC), and to provide an evidence for tissue engineering of vascular prosthesis., Methods: Bovine jugular veins were obtained fresh from a local slaughterhouse and were stored in chilled PBS. In the laboratory, any fat and loose connective tissue on the outer surface of the vessel was trimmed. BJVCs were decellularized by a 3-step extraction method as detergent Triton X-100 (0.5%), Trypsin (0.025%) EDTA (0.02%), and DNase I(30kU/L) RNaseA(0.3g/L). Histological and transmission electron microscopy (TEM) techniques were used to study the framework constituents of extracellular matrix of treated the examples, and fresh tissues were used as controls. Tissue contents of hydroxyproline(alkaline hydrolysis method) and elastin (Fastin Elastin Assay) were assayed respectively in the fresh and decellularized groups (n=10). The vascular wall heat shrinking temperature and mechanical strength were measured to evaluate the tissue stability (n=10)., Results: Histochemical and TEM analysis of BJVCs treated with decellularization proved a complete removal of nuclear and other cell components. Tissue collagen was well kept,but elastin was partly lessened. Tissue content of hydroxyproline increased comparatively [(25.73+/-2.97)mg/g vs. (29.25+/-2.99)mg/g, P<0.05] and the elastin content obviously decreased [(159.71+/-21.06)mg/g vs. (134.91+/-35.40)mg/g, P<0.05] in the decellular treatment group compared with the control group. The heat shrinking temperature and tensile stress of decelluarized tissue were lower than those of the fresh tissue[(72.50+/-0.53) degrees C vs. (69.75+/-0.54)degrees C ,P<0.05], [(5.19+/-0.65)MPa vs. (3.13+/-0.94)MPa, P<0.05]., Conclusion: The basic framework of extracellular matrix in the decellularized BJVC is partly damaged and tissue stability is reduced. Decellularized BJVC should be further crosslinked before being used as a tissue engineering scaffold for clinical pulmonary artery graft.

Objective: To summarize the surgical treatment of total anomalous pulmonary venous connection (TAPVC)., Methods: We retrospectively analyzed 49 patients with TAPVC as follows: 37 patients with supra-cardiac type in which 35 received anastomoses between the rear wall of the left atrium and the common pulmonary vein and the other 2 received anastomoses between the rear edge of the left atrium roof and the common pulmonary vein; another 12 patients with cardiac type who were incised the upper edge of coronary sinus, connected the common junction of pulmonary veins with the left atrium to form a new left atrium, and repaired the atrial septal defect with a pericardial patch., Results: Forty-six patients recovered and 3 patients died. Thirty-eight patients were followed-up from 3 months to 8 years. The rest patients got fluent pulmonary vein drainage and their heart function resumed to NYHA I., Conclusion: TAPVC patients should be operated on immediately at definite diagnosis. The fluency of the common pulmonary vein-left atrium anastomoses and proper postoperative care can ensure a satisfactory outcome.

Objective: To observe the structural and hemodynamic changes after the reconstruction of right ventricular outflow tract (RVOT) with valved bovine jugular vein conduit (BJVC) in the canine model by color Doppler echocardiography (UCG)., Methods: BJVC was used in the reconstruction of RVOT of 32 canines. UCG was used to observe the structure of the right ventricle and RVOT. The structure and hemodynamics were observed, and the diameter, width, blood velocity and pressure gradient (PG) were measured between the right ventricle and the conduits., Results: In the first stage, UCG showed that PG between the right ventricle and BJVC was high in 5 canines, and a few anastomosis in BJVCs with main pulmonary artery was small. Blood velocity quickened and PG increased, and color Doppler showed that the bloodstream was very bright. In the second stage, 7 canines survived for 1 year after the implantation. One year after the operation, UCG showed that the valve closed well, no graft kinking or obvious regurgitation of the valve was observed. The PG of the valve was low. The BJVCs were unobstructed and there was no thrombus.All the valves of BJVC opened well except one. In the third stage, 20 canines survived for 1 year after the implantation. UCG showed that the valve motion was good, no graft kinking or obvious regurgitation of the valve was observed. No neoplasm was observed., Conclusion: UCG is one of the most useful measurements in the structural and hemodynamic study of RVOT reconstruction by BJVC in the canine model. The good evaluation of UCG implies that it is important for the study and clinical practice.

Objective: To summarize the experience in treatment of Eisenmenger's syndrome by heart-lung transplantation (HLT)., Methods: Two cases of congenital heart disease with Eisenmenger's syndrome, aged 20 and 22, underwent bicaval orthotopic HLT. Modified St. Thomas cardioplegic solution and modified LPD solution were used to preserve the donor heart and lung. After removing the heart and lung of the recipient and thorough hemostasis in thoracic cavity and mediastinal septum, the donor heart and lung was implanted, and trachea, superior/inferior vena cava and aortic artery were anastomosed gradually. Intensive care against infection, rejection and other complications were performed after operation., Results: The operations were successful. Tracheal anastomotic stenosis happened in one case seven months after HLT and was cured by sleeve resection of the stenosed trachea segment. Acute rejection happened in one case 10 days after HLT and was cured by stoss therapy. Both patients recovered to normal life and work., Conclusion: The success of HLT is related to perfect organ preservation, precise surgical performance and proper peri-operative treatment.

Objective: To analyze the multiple factors affecting the postoperative mechanical ventilation supporting time in infants less than 10 kg with simple congenital heart diseases and to seize time by the forelock of extube and improve the outcome of surgical treatment., Methods: Data of 231 infants less than 10 kg with atrial septal defect(ASD),ventricular septal defect, and combining patent ductus arteriosus were retrospectively analyzed. The multivaricate stepwise logistic regression statistics were done for the predisposing factors affecting the ventilative supporting time., Results: The ventilative supporting time was 3~375 (average 23.5 h) h. The multivaricate stepwise logistic regression analysis indicated that severe pulmonary hyperpressure, cross-cramp aortic time, cardiopulmonary bypass time, preoperational pulmonary infection, membrane oxygenator, modified ultrafiltration, weight, and postoperative complications were significantly correlated to the ventilative supporting time., Conclusion: Severe pulmonary hyperpressure, preoperational pulmonary infection, long cross-cramp aortic time, long cardiopulmonary bypass time, postoperative complications all prolong the ventilation supporting time; the use of membrane oxygenator and modified ultrafiltration during the operation and big weight can diminish the pulmonary complications and shorten the ventilation supporting time.

Objective: To evaluate the angiogenic effect of the bone marrow mesenchymal stem cells (MSCs) transfected with human vascular endothelial growth factor (VEGF(165)) gene on myocardial infarcts in rats., Methods: The animal model of heart ischemic was established by ligating the left anterior descending coronary artery in Wistar rats. The ligated rats were divided into 4 groups (n=12 each), and 2 weeks later they were injected hVEGF-transfected MSC at the heart infarct zone (Group A), MSC (Group B), liposome-hVEGF gene plasmid (Group C), and medium (Group D). Four weeks after the injection, the capillary density of the infracted zone and the expression of human VEGF in vivo were examined., Results: Four weeks after the transplantation,the capillary density was significantly greater in Group A than that in Group B and Group D, slightly greater than that in Group C. The highest expression of hVEGF was Group A, and followed by Group C, Group B, and Group D., Conclusion: MSC is helpful for the stable expression of hVEGF gene, and is an ideal cellular vehicle for VEGF genes.

Objective: To determine the feasibility whether the bovine jugular venous conduit (BJVC) can be fixed with polyepoxy compound (PC)., Methods: Twenty-four BJVCs were divided into 3 groups and fixed with polyepoxy compound (PC group, n = 8), glutaraldehyde (GA group, n = 8), and unfixed group (Control group, n = 8), respectively. The morphologic and mechanical properties of BJVCs in the 3 groups, including thickness, diameter, moisture content, denaturation temperature, tensile strength, elongation at break, and fixation index were measured. The rat subcutaneous model for the assessment of tissue calcification was used. The calcium content in bovine jugular vein patches and valves was determined by flame atomic absorption spectrophotometer., Results: There was no difference in the wall thickness, diameter, and tissue water content between PC and the control group, but significant difference was found between GA and PC groups. The mechanical properties of PC group and GA group were not significantly different, but they were better than those of the control group. GA-fixed BJVC samples showed clear calcification, while PC fixed BJVC were calcified significantly less., Conclusion: PC is an effective and suitable choice for the treatment of BJVC since it can effectively preserve the structure and the anti-reflow function of valves in bovine jugular vein and it has better anti-calcification properties.

Objective: To create a method for transfecting human vascular endothelial growth factor165 (hVEGF165) gene into bone marrow mesenchymal stem cells (MSCs) in rats., Methods: MSCs of Wistar rats were isolated by density gradient centrifugation and purified based on their ability of adhesion to plastic. Detections of cell surface antigens, including CD34, CD45, CD44, and SH3, were performed using flow cytometry. MSCs' potential of differentiating into osteoblast and lipoblast in vitro was tested. The vector pcDNA(3.1)-hVEGF165 was transfected into MSCs with the liposome mediated method. The expression of hVEGF165 in the transfected cells was detected by enzyme linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR), and Western blot analysis., Results: The cultured MSCs were CD34-, CD45-, CD44+ , and SH+, which were differentiated into osteoblasts and lipocytes successfully. The expressed hVEGF165 in the transfected rat MSCs was demonstrated., Conclusion: The vector pcDNA(3.1)-hVEGF165 is successfully expressed in MSCs.

In 10 years after the interruption of filariasis transmission, the surveillance covered 29.5% of the townships and 3.7% of the population in the endemic areas, and 25.8% and 2.5% respectively after 10 years. No case with microfilaremia was found. 8248 floating people were also blood-examined with negative result. No filarial larva was detected in 103 261 Culex quinquefasciatus dissected. Out of 70,3498 people investigated in 708 villages of 249 townships, 667 chronic filariasis patients were found. Among 2928 people formerly with microfilaremia, no positive was found. The results indicate that Jiangxi Province has met the criteria of filariasis elimination set by the Ministry of Health.

Unlabelled: To establish a method to transfect vascular endothelial factor (VEGF) gene into mesenchymal stem cells ( MSCs) , to investigate the effects of the gene-transfected MSCs on heart function restoration and angiogenesis after myocardial infarction, and to compare the differences among cell therapy, gene therapy, and combined therapy., Methods: Seventy-one Wistar rats underwent ligation of the left anterior descending coronary artery so as to establish heart ischemia models. Fifteen rats underwent sham operation. MSCs were isolated from several Wistar rats by density gradient centrifugation, purified, and transfected with pcDNA3.1-hVEGF165 or blank plasmid pcDNA3.1 respectively using the liposome mediated method. ELISA, Western blotting, and RT-PCR were used to detect the protein and mRNA expression of hVEG in these MSCs Forty-eight surviving rats that underwent ligation were randomly divided into 4 equal groups: combination group (Combo group) to be injected into the heart infarct zone with suspension of hVEGF165-transfected MSCs 2 weeks after the establishment of the model, cell group to be injected with suspension of MSCs not transfected with VEGF, gene group to be injected with suspension of DNA-liposome containing pcDNA3.1-VEGF165 and control group to be injected with cold culture fluid only. Twelve surviving rats that underwent sham operation were used as normal non-ischemic group. Four weeks after the injection the surviving rats underwent examination of heart functions by the Buxco system. The rats were killed and their hearts were taken out to undergo immunohistochemistry with 5-bromodeoxyuridine (Brdu) and troponin T and factor VIII to measure the area of cardiac infarction and the capillary density. RT-PCR was used to examine the mRNA expression of VEGF. The heart infarcted size was calculated by Evan's blue staining., Results: (1) MSCs can be successfully isolated and cultured by density gradient centrifugation followed by adherence-separation. The expression of hVEGF165 in the transfected MSCs was demonstrated with ELISA, RT-PCR and Western Blot Assay. (2) Four weeks after the cells were transplanted, among all groups but the nonischemic group, the heart infarcted size of the Combo group was 27.8% +/- 3. 0% ,significantly less than those of the cell group (37.0% +/- 10. 1% ) and gene group (37.1% +/- 5.2%, both P <0.05). The heart function of the Combo group was better than those of other groups. (3) The capillary density of the Combo group was 40. 2 +/- 5.5/visual field, significantly greater than those of both the cell group (27.2 +/- 6. 3/visual field, P <0. 01) and that of the control group (18.5 +/- 5.8/visual field, P <0. 01) , and greater to some degree than that of the gene group (35. 8 +/-7.7/visual field, P =0. 189). (4)The heart infarcted size, heart function and capillary density of the cell and gene groups were similar and were smaller, better and greater than those of the control group. (5) Brdu and troponin T double staining detected a varied increase in the number of surviving cardiomyocytes at the heart infarcted area, some of which were double stain positive. RT-PCR showed mRNA expression of hVEGF165 in the Combo and gene groups, that in the Combo group being higher than that in the gene group., Conclusion: Eukaryotic expression vector pcDNA3.1-hVEGF165 can effectively be expressed in MSCs. Transplantation of VEGF gene by means of transfected MSCs brings better improvement in myocardial perfusion and in restoration of heart function than either cellular or gene therapy alone.

Post-traumatic inflammation response has been implicated in secondary injury mechanisms after spinal cord injury (SCI). Interleukin-1 (IL-1) is a key inflammatory mediator that is increasingly expressed after SCI. The action of IL-1 is mediated through its functional receptor, type I interleukin-1 receptor (IL-1RI). However, whether this receptor is expressed after SCI remains to be elucidated. In the present study, the temporospatial expression of IL-1RI was detected in rats that received a moderate contusive SCI (a 10 g rod dropped at a height of 12.5 mm) at the ninth to tenth thoracic vertebral level using a widely used New York University impact device. Our study demonstrated that IL-1RI was slightly increased at 4 h post-injury compared to the normal or sham-operated controls, reached the peak at 8 h at mRNA level (4.44-fold, P<0.01) and 1 d at protein level (2.62-fold, P<0.01). IL-1RI remained at its elevated levels for a relatively long duration (4 h-7 days). Spatially, IL-1RI was observed throughout the entire length of a 10 mm-long cord segment containing the injury epicenter. Colocalization of IL-1RI was found in neurons, oligodendrocytes, astrocytes, and activated microglia. Our results suggest that the elevated expression of IL-1RI after SCI may contribute to posttraumatic inflammation responses of IL-1.

The biological effects of interleukin (IL)-1 are mediated by two distinct receptors, the p80 or type I (IL-1RI) and p68 or type II (IL-1RII) receptors. Because IL-1RII has a short, 29-amino acid cytoplasmic domain which may not be sufficient for signaling, there is considerable evidence indicating that IL-1 may signal exclusively through the IL-1RI receptor. Here, we report the expression, distribution, and cellular localization of the IL-1RI protein in the adult rat spinal cord in vivo and embryonic spinal cord in vitro. We found that IL-1RI was expressed in both the gray and white matter throughout the entire length of the spinal cord and was localized in neurons of the anterior horn, astrocytes, oligodendrocytes, and central canal ependymal cells. Interestingly, resting microglia were negative for IL-1RI. In primary cultures obtained from the embryonic day (E) 15 rats, IL-1RI was expressed in neurons, astrocytes, and oligodendrocytes as well as microglia. These data provide both in vivo and in vitro evidence that neurons and glial cells express the IL-1RI proteins. The differential expression of IL-1RI in the developing, but not mature, microglia may indicate the difference of these cells in response to IL-1 stimuli during maturation. The distribution and cellular localization of IL-1RI proteins in the spinal cord provide a molecular basis for understanding the reciprocal interaction between the immune and the central nervous systems.

Objective: To determine the lung protection of continuous pulmonary artery perfusion with oxygenated blood during cardiopulmonary bypass (CPB)., Methods: Thirty patients undergoing mitral valve replacement were randomly divided into the control group (n=15) and the lung perfusion group (n=15). The patients in the lung perfusion group were perfused oxygenated blood continuously to the pulmonary artery during CPB. The patients in the control group were performed the routine procedure of mitral valve replacement. Record the CPB time, aortic cross-clamp time, mechanical ventilation time and ICU monitoring time. The patients' oxygen index (OI) and lung static compliance (Cstat) were measured before the surgery, at 0 h after the CPB and at 0, 6 h after the surgery. Right lung biopsy specimens were obtained at 30 min after the CPB to observe the histological changes. Results The mechanical ventilation time and ICU monitoring time were shorter in the lung perfusion group than those in the control group (P < 0.05). The patients' OI and Cstat were higher after surgery in the lung perfusion group than those in the control group (P < 0.05). Tissue examination showed lung parenchyma edema and inflammatory cells accumulated in the control group, while no remarkable pathological changes occurred in the lung perfusion group., Conclusion: Lung injury exists after the surgery by CPB. Continuous pulmonary artery perfusion with oxygenated blood during CPB can decrease the lung injury.

Objective: To summarize the experience in prosthetic valve replacement in pediatric patients., Methods: The clinical data of consecutive 105 children, 63 male and 42 female, aged 13 +/- 3 (1.5-16), with the underlying diseases of rheumatic heart disease (n = 55), congenital heart diseases (n = 43), and infective endocarditis (n = 5), with the preoperative cardiac function of class II (n = 36), or class III or IV (n = 69), who underwent prosthetic valve replacement, including replacement of mitral valve (n = 58), aortic valve (n = 28), tricuspid valve (n = 5), mitral and aortic valves (n = 13), and mitral and tricuspid valves (n = 1), and tricuspid valvuloplasty (n = 26), from May 1984 to May 2004, were respectively analyzed. Mechanical valves were used and took low dose anticoagulant warfarin was administrated to all cases. Postoperatively prothrombin time (PT) and international normal ratio (INR) were observed. Follow-up lasting 10 +/- 4 years (6 months to 20 years) was performed among 100 discharged patients with a follow-up rate of 91.0%., Results: There were 5 early deaths (4.8%), mostly owing to serious low cardiac output syndrome, and 4 late deaths owing to endocarditis (n = 2), heart failure (n = 1), and serious arrhythmia (n = 1). The 87 surviving patients showed their cardiac function of class I-III. No serious complications related to anticoagulation and prosthesis dysfunction had been found., Conclusion: Prosthetic valve replacement with mechanical valve can be performed in pediatric patients with good outcome. Larger type of mechanical valve should be used as possible. Additional surgical procedures may be required to correct the concomitant malformations during the operation. Postoperative long-term low dose anticoagulant treatment is safe.

Objective: To study the surgical treatment effect and factors influencing prognosis of recurrent carcinoma within gastric remnant., Methods: From 1990 to 2003, clinical data of 60 patients underwent surgical operation again because of recurrence carcinoma within gastric remnant after radical resection were reviewed retrospectively., Results: The mean survival time was 32.6 months and the median survival time was 27.0 months. Thirty-seven patients (61.7%) received radical resection with the mean survival time of 45.0 months. Twenty-three patients received palliative treatment with the mean survival time of 12.7 months (P< 0.001). Univariate analysis showed that the clinical stages, pathologic classification, radical resection, peritoneal seeding, liver metastasis, the expression of CD44v6, PCNA and MVD were significantly correlated with survival time (P< 0.01), but were not correlated with sex,age,recurrence time after the first operation (P> 0.05). Multivariate analysis revealed that the clinical stages, peritoneal seeding and radical resection were independent prognostic factors., Conclusions: The clinical stages, peritoneal seeding and radical resection are independent prognostic factors of recurrent carcinoma within gastric remnant. The radical resection maybe the most effective way to treat the recurrent carcinoma within gastric remnant.

Objective: To summarize the experience of surgical treatments of partial atrioventricular septal defect in 60 patients., Methods: From April 1999 to April 2004, 60 patients of partial atrioventricular septal defect were operated. Fifty-eight patients were performed with suture of the cleft of mitral valve and the other 2 were given mitral valve replacement; For closure of primum ASD, 53 patients with pericardial patches and 7 with Dacron patches. Coronary sinus was baffled to left atrium with kirklin procedure in 35 cases and baffled to right atrium with McGoon procedure in other 25 cases. Correct the accompanying cardiac deformity at the same time., Results: The hospital mortality was 3.3% (2/60) due to low cardic output syndrome. The incidence rate of complete atrioventricular block was 8.00% (2/ 35) in the group with Kirklin procedure and 6.06% (2/25) in the group with McGoon procedure. There was no statistical significance between the 2 groups (P > 0.05). The follow-up was from 1 month to 5 years, and there was no late death. All cardiac function were improved except middle mitral regurgitation in 1 patient., Conclusion: Reasonable operative design, refined procedures, avoiding damage to conducting bundles and proper perioperative management are the key points in improving theraeutic effect.

We have previously established a culture method to isolate and cultivate neural stem cells (NSCs) derived from the rat embryonic brain and spinal cord. In the present study, we demonstrate that the spinal cord-derived NSCs can be induced to differentiate into oligodendrocyte precursor cells (OPCs) with a combined treatment composed of (1) conditioned medium collected from B104 neuroblastoma cells (B104CM) and (2) basic fibroblast growth factor (bFGF, 10 ng/ml). After induction, over 95% of the cells displayed bipolar or tri-polar morphology and expressed A2B5 and platelet derived growth factor receptor-alpha (PDGFR-alpha), markers that are specific for OPCs. Among PDGFR-alpha positive OPCs, only a few cells expressed glia fibrillary acidic protein (GFAP) and none expressed beta-tubulin III. In the presence of B104CM and bFGF, OPCs proliferated rapidly, formed spheres, expanded for multiple passages, and maintained their phenotypic properties. Upon withdrawal of B104CM and bFGF, these cells differentiated into either O4/GlaC-positive oligodendrocytes (OLs) or GFAP- and A2B5-positive type-2 astrocytes. Our results indicate that NSCs can be induced to differentiate into OPCs that possess properties of self-renewal and differentiation into oligodendrocytes and type-2 astrocytes, a property similar to that of O-2A progenitor cells. The OPCs can be maintained in an undifferentiated state over multiple divisions as long as both B104CM and bFGF are present in the medium. Thus, large quantity of OPCs can be obtained through this method for potential therapeutical interventions for various neurological degenerative diseases.

Objective: To summarize the experience of diagnosis and surgical treatment of ventricular septal defect with patent ductus arteriosus., Methods: We retrospectively analyzed the clinical data of 102 cases of ventricular septal defect combined with patent ductus arteriosus who underwent surgical treatment. Preoperative ultrasonic cardiogram (UCG) showed ventricular septal defect combined with patent ductus arteriosus in 82 cases and ventricular septal defect in 20 cases., Results: The hospital mortality was 4.9% (5/102). The reasons for death included low cardiac output syndrome (1 case), pulmonary hypertension crisis (2 cases) and respiratory failure (2 cases). In the remaining patients,the perioperative complications included lung infection (7 cases), pulmonary atelectasis (5 cases), hydrothorax (1 case), and pulmonary hypertension crisis (2 cases); and all the 15 patients recovered lastly. The pulmonary hypertension of all living patients decreased to some degree. The therapeutical effectiveness was satisfactory., Conclusion: Ventricular septal defect with patent ductus arteriosus is easy to be confused with ventricular septal defect clinically. At the same time,it is diffcult to form a correct diagnosis in some patients by UCG preoperatively. To prevent the occurrence of perfusive lung, it is important to reinforce preoperative diagnosis and exploration during operation. Because pulmonary hypertension in patients with ventricular septal defect with patent ductus arteriosus emerges early and develops quickly, it tends to result in organic pulmonary hypertension which can make patients lose operation chances and influence the long-term therapeutical effect. Surgical operation should be performed as soon as possible. Optimal operative timing and proper perioperative management play important roles in surgical results.

Objective: To determine the application of modified low-potassium dextran (LPD) solution in heart-lung transplantation., Methods: We used the modified LPD as the lung flush solution of the donor in the first heart-lung transplantation procedure in Hunan., Results: The patient survived 555 days after the surgery and severe infection, lung disfunction or severe graft rejection never occurred., Conclusion: Modified LPD as lung flush solution of the donor during heart-lung transplantation demonstrates an excellent effect of lung protection.

Objective: To evaluate the immunogenicity of bovine jugular vein conduits (BJVCs) treated with different cross-linking methods., Methods: The BJVCs were treated with glutaradehyde (GA), dye-mediated photooxidation (DMP) and polyepoxy compound (PC) (n = 10). The tissue homogenates obtained from BJVCs treated with PC, GA, DMP, and fresh BJVCs, were mixed with the complete Freand adjavant to form the emulsive antigen, which were used to immunize rabbits correspondently. The antibody concentrations to BJVCs in those rabbits' serum were measured by counter double immuno diffusion. The immunologic responses to the BJVCs in different groups were measured with Western blotting., Results: The positive bands appeared when the sera of rabbits were immunized by fresh BJVCs reacted with antigens of fresh BJVCs, but no bands appeared when the sera of rabbits were immunized by fresh BJVCs reacted with those antigens of the BJVCs treated with GA, DMP, and PC in Western blotting., Conclusion: The immunogenicity of BJVCs treated with PC, DMP, and GA can be reduced significantly and meet the clinical standard.

Objective: To investigate the effect of pretreatment with captopril on myocardium ischemia-reperfusion injury in atherosclerotic rabbits., Methods: Thirty-two New Zealand white rabbits were assigned randomly to the normally feed group, cholesterol-feed (CF) group, and cholesterol food plus captopril group (cap-feed group), which were fed for 10 weeks. We examined the changes in the size of the infarct and changes in the myocardium ultrastructure resulting from coronary ischemia/reperfusion. Levels of endothelin (ET) and nitic oxide (NO) were measured in the different experiment stages., Results: The ET levels significantly increased and the content of NO significantly decreased in the CF group compared with those of the cap-feed group. The ultrastructure of myocardium cell was slightly destroyed and the infarct size was significantly smaller in the cap-feed group than the normally feed rabbits and CF rabbits., Conclusion: The long-term captopril treatment can lighten the severity of myocardial injury produced by coronary ischemia/reperfusion.

Aim: To explore the role of MAPK signal transduction pathway in killing tumor cells by Mycobacteria tuberculosis antigen (Mtb-Ag) activated human gammadeltaT cells., Methods: Normal human peripheral blood mononuclear cells (PBMCs) were stimulated with Mtb-Ag and expanded in rIL-2-containing medium to induce gammadeltaT cells. The highly purified gammadeltaT cells were isolated by positive selection with MACS separator. Proportion of gammadeltaT cells was detected by flow cytometry (FCM). The cytotoxicity and CD69 expression of gammadeltaT cells pre-treated with or without PD98059 (Erk inhibitor) or SB203580 (p38 inhibitor) were detected by MTT colorimetry assay and FCM, respectively., Results: The percentages of gammadeltaT cells in freshly isolated PBMCs, Mtb-Ag-activated PBMCs and cells sorted by MACS were 3.56%, 74.63% and 98.20%, respectively. The cytotoxicities of gammadeltaT cells to tumor cell lines (K562 and Raji cells) were inhibited by PD98059. The inhibitory rate of cytotoxicity to K562 cells by gammadeltaT cells (39.27%) was higher than that to Raji cells(26.58%). CD69 expressions on gammadeltaT cells induced by K562 or Raji cells were decreased by pretreatment with 100 mumol/L of PD98059. But SB203580 had no effects on the cytotoxicity and CD69 expression on gammadeltaT cells induced by K562 and Raji cells., Conclusion: Erk MAPK pathway, not p38 MAPK, plays an important role in triggering cytotoxicity of gammadeltaT cells. The Erk pathway may affect on the granule exocytosis of gammadeltaT cells more than the Fas/FasL-mediated cytotoxicity of gammadeltaT cells. Moreover, Erk MAPK pathway is also involved in expression of activation molecule CD69 on gammadeltaT cells induced by tumor cells.

Growth inhibition in the central nervous system (CNS) is a majar barrier to axon regeneration. To date, three inhibitory proteins of oligodendrocytes/myelin have been identified: myelin associated glycoprotein ( MAG) , Nogo-A and oligodendrocyte-myelin glycoprotein (OMgp). Recently, findings indicate that three distinct myelin components inhibit axon growth by binding a common receptor, the Nogo 66 receptor (NgR). These novel findings lead to further understanding of the barriers to axon regeneration after injury in the CNS and provide new avenues to explore the new theraphy strategy for the CNS injury.

Objective: To provide some references for defining the Chinese optimal intensity of anticoagulation after mechanical heart valve prostheses replacement., Methods: For the 178 patients with carbomedics mechanical prosthetic heart valves, the means of INR were compared between the patients with complications and those without complications at the standard of INR1. 4 - 2.0. Also, the variations of INR were compared among different follow-ups., Results: During the follow-up, 22 hemorrhagic and 1 thromboembolic complication occurred. The total linearized rate of anticoagulation-related hemorrhage was 5.83% pty. The total linearized rate thromboembolism was 0.26% pty. The late mortality was 0.79% pty (3 cases ). The final mean INR was 1.68 +/- 0.38. The final mean oral warfarin dose was 2.34 +/- 0.80 mg. The differences of variations of INR in five periods were significant (F = 5.072, P < 0.05). The mean INR in the first month of follow-up was 1.75 +/- 0.27., Conclusion: For Chinese patients with mechanical prosthetic heart valve, hemorrhage is the principal complication, the ratio of which is much higher than that of thromboembolism. The low-dose anticoagulation (INR1. 4-2.0) could remarkably decrease hemorrhagic events as effectively as prevent the thrombolic events. Moreover the INR is the most unstable in the first month of follow-up, so re-examination for the patients in the first month after the operation is vitally important.

Objective: To evaluate the expression and myocardial protection of heat shock protein-70 (HSP70) in the myocardial cells of cyanosis congenital heart diseases., Methods: The study enrolled 24 patients with congenital heart disease: 12 cases of non-cyanosis (group A), and the other 12 cases of cyanosis (group B). During the operation the samples of myocardium in the right atrium were attained at 0 min ischemia, 20 min ischemia, and 20 min reperfusion. They were used to evaluate the content of HSP70 by immunohistochemistry and activation of SOD and content of MDA by chemochromatometry., Results: The content of HSP70 in myocardial cells during the operation was not significantly different in both groups (P > 0. 05). The content of HSP70 significantly decreased in group B compared with that of group A (P <0.05). The activity of SOD significantly decreased and the content of MDA significantly increased in group B during the operation (P < 0.05)., Conclusion: Decreased expression of HSP70 is associated with the decreased tolerance to ischemia-reperfusion injury in myocardial cells of cyanosis congenital heart disease.

Objective: To investigate the effect of cardiac transplanting of mesenchymal stem cells on the apoptosis after rat myocardial infarction., Methods: Wistar inbred rats were used to stimulate autograft. Harvested rat mesenchymal stem cells were cultivated, proliferated and labeled in vitro. At the same time, myocardial infarct models were set up using liquid nitrogen to cryoinjury the free wall of the left ventricule. Four weeks later, 2 x 10(6) mesenchymal stem cells or cold D-Hanks liquid were injected into several different points of infarct area. After 1, 2 and 4 weeks, specimens from infarct area were obtained in order. Then the number of apoptosis was counted under scanning electron microscope in every 2 specimens selected at random from the experimental group and the reference group within 2 weeks. The bcl-2 mRNA expression of all rats was assayed by RT-PCR., Results: The observation of transmission by electron microscope showed that part of the myocardial cells became apoptosis in the initial stage in ultramicrostructure. Apoptosis ratio was 0.02 and 0.08 in the experimental group and the reference group respectively, which demonstrated that mesenchymal stem cells could relieve apoptosis after rat myocardial infarctation. The levels of bcl-2 mRNA expression in 1, 2, and 4 weeks were all obviously promoted in the experimental 1.2. group (3.235 +/- 0.126, 1.152 +/- 0.021, 0.798 +/- 0.016) than those in the reference group (2. 695 +/- 0.084, 0.537+/-0.022, 0.579+/-0.019) (P < 0.01), and quickly decreased with time., Conclusion: Mesenchymal stem cells transplantation can promote the expression of bcl-2 mRNA in the infarct area and relieve apoptosis after rat myocardial infarction.