The effects of oxidized low density lipoprotein (ox-LDL) on the proliferation of cultured human vascular smooth muscle cells (vSMC) were investigated in vitro. By using NaBr density gradient centrifugation, LDL was isolated and purified from human plasma. Ox-LDL was produced from LDL by being incubated with CuSO(4). ox-LDL was then added to the culture medium at different concentrations (35, 60, 85, 110, 135 and 160 microg/mL) for 7 days. The influence of ox-LDL on vSMC proliferation was observed in growth curve, mitosis index, and in situ determination of apoptosis. The data were analyzed with SPSS 10.0 software. The results showed that the ox-LDL produced in vitro had a good purity and optimal oxidative degree, which was similar to the intrinsic ox-LDL in atherosclerotic plaque. ox-LDL at a concentration of 35 microg/mL demonstrated the strongest proliferation inducement, and at a concentration of 135 microg/mL, ox-LDL could inhibit the growth of vSMC. ox-LDL at concentrations of 35 and 50 microg/mL presented powerful mitotic trigger, and with the increase of ox-LDL concentration, the mitotic index of vSMC was decreased gradually. ox-LDL at higher concentrations promoted more apoptotic vSMCs. ox-LDL at lower concentrations triggered proliferation of vSMCs, and at higher concentrations induced apoptosis in vSMCs. ox-LDL played a promotional role in the pathogenesis and development of atherosclerosis by affecting vSMC proliferation and apoptosis.

A promising genetic marker, sag5b, was cloned and expressed and the difference of the genes between highly virulent strain (RH) and less virulent strain(Prugniaud) of Toxoplasma gondii was compared. The PCR-generated product of sag5b was subcloned into T easy vector and plasmid pET28a consecutively. The fusion expression was induced by IPTG and identified by SDS-PAGE and Western blotting. The immunoreactivity of recombinant SAG5B was identical to that of native SAG5B on the membrane of tachyzoites of RH strain. The brains of mice infected with Prugniaud strain of T. gondii were homogenated. Sag1 was successully cloned by PCR from both RH strain tachyzoites and the homogenized brain tissues of mice infected with low virulent strain of Prugniaud,whereas sag5b was only detected in RH strain but not in Prugniaud strain, indicating that sag5b could be used as a genetic marker for differentiation of strain virulence. Expression and vaccination of the virulence-associated gene into mice failed to induce obvious protective immunity against the challenge of RH strain.

Objective To isolate specific humanized anti-D-dimer scFv(single chain Fv) antibody from scFv phage libraries. Methods Isolate anti-D-dimer positive clones from Tomlinson I + J phage libraries by three rounds of panuing, then sequence monoclonal genes by bideoxy-mediated chain termination and express soluble scFv antibody; Pick out anti-D-dimer antibodies with high specificity and affinity by ELISA.Results After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer scFv fragments were selected. By polyclonal and monoclonal phage ELISA and gene sequencing, 20 different full-length monoclonal scFv phages were identified, the result of soluble scFv ELISA showed that 20 full-length monoclonal scFv were expressed smoothly. According to the result of soluble scFv ELISA, in 5 scFv antibodies with high value of A450 selected, 3 scFv antibody fragments showed high specific and affinity. Conclusion Antibody phage display was an effective, rapid method to isolate anti-D-dimer antibodies with high specificity and affinity.

Objective: To evaluate the safety and efficacy of nonmyeloablative preconditioning allogeneic hematopoietic stem cell transplantation (NMHSCT) in the therapy of unidentified relapse and primary solid tu-mors, and to study the anti-tumor immunity induced by the effect of Graft-Verus-Tumor (GVT). Methods: A to-tal of 13 difficult-to-treat cancer patients received NMSCT and the efficacy and side effects were observed. Re-sults: One case had CR, 2 cases had PR, 4 cases had SD, 5 cases had PD, and 1 case died of complica-tions associated with transplantation. One case was GVT (+++), 3 cases were GVT (++), 5 cases were GVT(+), and 4 cases were GVT (-). The main side effect was acute GVHD presented as diarrhea and infec-tion. VOD and brain disease were rare. Conclusion: Nonmyeloablative allogeneic stem cell transplantation is safe and effective for solide tumors.

Animals ; Brain Chemistry ; Brain Ischemia ; metabolism ; physiopathology ; Cerebral Cortex ; metabolism ; Dementia ; metabolism ; physiopathology ; Humans ; Microdialysis ; Reperfusion Injury ; metabolism ; physiopathology

Aged ; Humans ; Inflammation ; Laryngeal Diseases ; Laryngeal Neoplasms ; Male ; Neoplasms, Muscle Tissue

Objective: To study the effect of enteral nutrition in the treatment of heart failure with hyponatremia.Methods: Forty patients sufferd from heart failure with hyponatremia were divided into two groups at random: control group(n=20) with regular therapy and treatment group(n=20) with enteral nutrtion on the base of regular therapy.Time intervals from hyponatremia to normal natremia of the two groups were compared.Results: Time interval from hyponatremia to normal in the treatment group was 5.30?1.82 days,and that in the control group was(8.85?2.44) days.Conclusion: The treatment with enteral nutrition can significantly shorten the clinical treatment time in heart failure with hyponatremia.

Around the core issue how to evaluate and test hypothesis, starting from the concept of hypothesis and scientific hypothesis, learning form recent progress and understanding at home and abroad, we have analyzed the type of scientific hypothesis, test methods and pre-mortem and post-mortem evaluation criteria in depth, established scientific vision and discipline ruler, explored the awareness level of the Chinese medicine hypothesis, pointed out the problems and gaps. Whereby, we have analyzed and demonstrated brain regulating five internal organs concept of wholism scientific hypothesis, and proven it belongs to the scientific hypothesis. We were confident that these works mentioned above would lead the theoretical reform changing Chinese medicine hypothesis from traditional description to the nature clarification.

To investigate the expression of osteopontin (OPN) and its receptor integrin alphanubeta3 in the placental tissue from pregnant women complicated with preeclampsia, the expression of OPN and alphanubeta3 in the placenta of the pregnant women with preeclampsia and healthy pregnant women was detected by immunohistochemistry, Western blotting and RT-PCR. Our results showed that OPN and alphanubeta3 protein were expressed in the placenta from normal pregnant woman and those with preeclampsia. OPN was located in the placental syncytiotrophoblasts and the cytoplasm of capillary endothelial cells and integrin alphanubeta3 was mainly expressed on the surface of trophoblast cells. Expression of OPN and integrin alphanubeta3 in the placental tissue from preeclampsia subjects was significantly lower than that from the control group (P<0.05). Compared with the control group, expression of OPN in the placental tissue from preeclampsia group was significantly lower (P<0.05) but there was no significant difference in the expression of alphanu and beta3 between the preeclampsia group and the controls. It is concluded that OPN and its receptor integrin alphanubeta3 may be involved in the pathogenesis of preeclampsia.

Long-term treatment with an agonist of peroxisome proliferator-activated receptor (PPAR)-γ is associated with bone fractures in the clinical practice. However, the mechanisms underlying the fractures are not fully understood. This study was aimed to examine the effect of rosiglitazone (an agonist of PPAR-γ) of different doses on the proliferation, differentiation, and transforming growth factor beta 1 (TGF-β1)-induced expression of connective tissue growth factor (CTGF) in primary rat osteoblasts in vitro. Osteoblasts were isolated from newly born SD rats and treated with different doses of rosiglitazone (0-20 μmol/L). The proliferation and differentiation of osteoblasts were measured by MTT assay and NPP assay, respectively. The expression of CTGF was determined by RT-PCR and Western blotting. The results showed that most isolated osteoblasts displayed strong alkaline phosphatase (ALP) activity and treatment with different doses of rosiglitazone did not affect their proliferation, but significantly inhibited the differentiation of osteoblasts in a dose-dependent manner. Moreover, treatment with different doses of rosiglitazone significantly reduced the TGF-β1-induced CTGF mRNA transcription and protein expression in a dose-dependent manner in rat osteoblasts. It was concluded that the activation of PPAR-γ may inhibit the differentiation of osteoblasts by reducing the TGF-β1-induced CTGF expression in vitro.