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 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.

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.

Objective To study the clinical characteristics and outcome of cytomegalovirus(CMV) infection in infants.Methods All data including time of infection,clinical characteristics,laboratory tests and outcome of CMV infections in hospitalized infants were collec-(ted) and analyzed from January,1994 to July,2004.Results In 87 infected infants,congenitally infected newborns,perinatal infection in infants and postnatal infection in infants accounted for 27.6%,62.0%,16.6%,respectively.CMV hepatitis was the most frequent type of disease with the incidence of 41.3%,in which the incidence of splenomegaly was 10.3%.Most of CMV hepatitis infants had a good prognosis with the improved rate 80.5%.Central nervous system abnormality(including abnormal intension of muscle,convulsion,ocular and hearing abnormalities) occurred only in congenital and perinatal infection with the incidence of 20.4%.Generalized infection,the incidence of congenital infection and perinatal infection was 16.7%,1.8%,respectively.It did not occur in postnatal infection.The mortality rate of congenital infection and perinatal infection were 12.5% and 1.85%,respectively.Conclusions CMV infection is the main cause of infant hepatitis and it can also cause neurologic sequelae.The outcome of generalized infection in congenital infection is bad and the mortality rate is high.

Objective To analyze the expression of phosphorylated protein kinase B(pAKT)and PTEN protein in ovarian epithelial cancer and to investigate the correlations between their expression and prognosis of ovarian epithelial cancers.Methods Expression of pAKT and PTEN in 12 normal ovarian tissues,20 benign tumors,12 borderline tumors and 80 cases of ovarian epithelial cancers were detected by immunohistochemical method,and their correlations were analyzed.Results The positive expression of pAKT in normal ovarian and benign tumor tissues were significantly lower than that in ovarian epithelial cancers(8%,10% vs 55%;P

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.

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

Objective To investigate peripheral blood T lymphocyte subsets and NK cells changes in the diffuse large B cell lymphoma (DLBCL) patients before and after chemotherapy,and analyse the relationship between the results and treatment.Methods Collect the 47 patients venous blood of the effective treatment DLBCL by pathology.T lymphocyte subsets and NK cells were determined by flow cytometric.Analyse the results statistically significant difference before treatment,the second chemotherapy cycle and the fourth chenmotherapy cycle compared with 50 healthy control persons.Results The levels of CD3+,CD4+,CD4+/CD8+ NK cells in DLBCL patients before chemotherapy [(70.04±8.87)％,(42.79±6.06)％,(1.68±0.59)％,(14.40±6.02)％]were lower than healthy controls [(63.89±6.67)％,(32.72±5.77)％,(0.85±0.25)％,(9.95±5.24)％](P ＜ 0.05),and the level of CD8+ cells is higher than the healthy controls [(27.21 ±6.54)％ vs.(39.92±7.11)％](P ＜ 0.05).The levels of CD3+,CD4+,CD4+/CD8+ cells had significant difference between the second and the fourth chemotherapy cycle in DLBCL patients (P ＜ 0.05).The levels of CD3+,CD4+,CD8+,CD4+/CD8+,NK cells had significant difference between the fourth chemotherapy cycle DLBCL patients and the DLBCL patients before chemotherapy (P ＜ 0.05).Conclusion The DLBCL patients exist immunosuppression before chemotherapy.Peripheral T lymphocyte subsets and NK cells can be used as good reflect of the cellular immune function in DLBCL patients.Clinical parameters can be used for the immune function monitoring and providing guidance for the treatment options.

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.