To reverse drug resistance mediated by the MDR1 gene product P-glycoprotein(P-gp) in tumor cells in a specific manner, a hammerhead ribozymes which can cleave the GUC sequence in codon 196 of MDR1 mRNA was designed and cloned into a recombinant retroviral vector pDOR-neo at BamH I restriction sites and packaged with packaging cell line PA317 cells.The viral supernatant was used to infect the multidrug-resistant human hepatocarcinoma cell line BEL-7402/DOX. After selection with G418,resistant colonies were obtained.Stable expression of retroviruses in both PA317 and infected BEL-7402/DOX cells was confirmed by Northern Blot hybridization. Down-regulation of P-gp and even of MDR1 mRNA was found in BEL-7402/DOX infected with ribozyme construct. The rate of BEL - 7402/DOX infected cell defected by flow cytometric analysis was 8.2 ~ 14.6% while in uninfected cell it was 93 .4 ~ 97.5% . The BEL-7402/ DOX cell infected with ribozyme construct was found back to drug-sensitivity to a series of drugs by MTT colorimetric assay . The results demonstrated that the recombinant retroviral vector expressing ribozyme transfecting human hepatocarcinoma BEL-7402/DOX could inhibit MDR1 gene expression and reverse tumor MDR phenotype back to drug-sensitive condition.

Objective:To study the effect of the cytokines, tumor necrosis factor-?(TNF-?), interferon-?(IFN-?), interferon-?(IFN-?), platelet-derived growth factor BB(PDGF-BB), basic fibroblast growth factor(bFGF),on promoter activity of human transforming growth factor-?1(TGF-?1) gene.Methods:A construct of phTGF2.14 containing sequence from -1 328 bp to +812 bp of human TGF-?1 gene,linking with chloramphenicol acetyltransferase(CAT) as reporter gene,was transiently transfected into rat hepatic stellate cell line nFSC. The cells were subsequently treated with TNF-?,IFN-?,IFN-?,PDGF-BB,bFGF, and the CAT activity was assessed 48 hours after stimulation with each cytokines.Results:TNF-? of 10 ng/ml can increased the CAT activity of phTGF2.14 to 3.24 fold compared to control. IFN-? and IFN-? at 1 000 U/ml decreased CAT activity to (42?12)% and (58?6)% of control respectively.PDGF-BB,bFGF of 10 ng/ml had no effect on promoter activity of human TGF-?1 gene;Combined application of IFN-?,IFN-? and TNF-?,the promoter TGF-?1 gene were 1.32 and 1.46 fold compared with control,respectively.Conclusion:These data indicate that TNF-? can increase the promoter activity of human TGF-?1 gene, but IFN-?,IFN? can downregulate the CAT activites of phTGF2.14, and IFN-?,IFN-? can interdict the upregulate effect of TNF-? on phTGF2.14. We did not find PDGF-BB,bFGF have any effect on TGF-?1 promoter. These provided an essential evidence for study the interaction mechanism of cytokines in fibrogenisis diseases.

[Objective]To test and prove the mushroom shaped surface prosthesis of femoral head not only can be a weight-bearing device,but also could reduce stress-shelter and achieve long-term stability.[Method]A simplified three-dimensional model of femoral head was made.And the biomechanics of femoral head were compared by finite element analysis in two states:with and without the replacement.[Result]It showed that the maximum of strees in famoral head was less than 40Mpa,consequently the replacement had sufficient strength to meet its basic function.Furthermore,the prosthesis bear the force both at up part and down.Therefore it reduced stress-shelter.[Conclusion]This prosthesis can achieve immediate and long-term stability.And it provides new ideas for designing the femoral head resurfacing prosthesis.

Objective To investigate the Th1/Th2 polarization of T lymphocytes in human peripheral blood stimulated by CpG-ODN2216,and the secretion of cytokines in supernatant of cultured PBMCs after stimulation of CpG-ODN2216.Methods Human PBMCs were isolated from blood of donors.The PBMCs were incubated with CpG-ODN2216 for 24 hours.Th1/Th2 subsets in the cultured PBMCs were examined by flow cytometry,and IFN-?,IL-2,IL-4 and IL-10 in the supernatant were assayed by ELISA.Results The percentage of Th1 and Tc1 increased significantly after stimulation of CpG-ODN2216 compared with control group (P0.05).Conclusion The Th1 cells and Tc1 cells in T lymphocytes of peripheral blood could be polarizated by CpG-ODN 2216.IFN-? secretion in PBMCs could be induced by CpG-ODN2216.

Objective To address the medical problems, in treatment of avascular necrosis of the femoral head, on loosening and subsidence of the prosthesis of femoral head. A set of Mushroom Shaped Surface Prosthesis of Femoral Head was designed. Methods Based on biomechanical of the femoral head and the principle of transferring the force through femoral head while reducing stress-sheltered, nickel-titanium alloy and other new medical alloy materials were used to develop a prosthesis, which not only can be a weight-bearing device, but also promotes bone-growth. Results The prosthesis is novel, compact and little-volume, and the manipulation of surgery is convenient and secure. Conclusion The design of this prosthesis can provide new ways or ideas for designing the femoral head replacement prosthesis to solve problems on loosening and subsidence.

Objective To investigate the prevalence of metabolic syndrome (MS) and its components among the elderly at Tanggu community in Tainjin. Methods The elderly inhabitants were drawn out randomly from one community at Tanggu in Tianjin to examine the MS-related indexes. There were 623 samples with complete data aged (67.4±6.0) years, including 261 males with mean age of (68.3±6.1) years and 362 females with mean age of (66.9±6.0) years. Diagnosis of MS was done by criteria of Chinese Medical Association Diabetes Branch in 2004. Results The standardized prevalence of MS for the elderly in the community was 18.5% with the highest risk group aged 60～69 years. Standardized prevalences of hypertension, high triglycerides, high blood sugar, overweight, obesity, low HDL-C were 49.5%, 38.8%, 25.3%, 42.0%, 4.2%, 3.0% in the elderly. Conclusions The prevalence of MS in the elderly is lower in Tanggu community in Tianjin than in Shanghai, Foshan in Guangdong, Qingdao in Shandong and Beijing city. The condition in female is more severe than in male, the highest prevalence occurs in hypertension, and overweight, high triglycerides and high blood sugar follow it, but few of them are obesity, low HDL-C is fewer too.

Objective To explain the physiological properties of serum leucine aminopeptidase (LAP) and discuss the different levels and clinical meaning of serum LAP during the second and the third trimester of gestation.Methods Rate turbidimetry was used to assay serum LAP activity of 175 pregnant women in the second trimester of gestation, among whom 162 were normal and 13 were intrahepatic cholestasis of pregnancy (ICP), and 302 pregnant women in the third trimester of gestation, among whom 215 were normal, 76 were ICP, 6 were pregnancy-induced hypertension syndrome and 5 were premature delivery, and 100 healthy or non-pregnant women.Results Serum LAP activity of pregnant women in the group of the second trimester of gestation was (140.43?64.39) U/L, in the group of the third trimester of the gestation was (234.72?93.53) U/L, and in control group was (72.62?15.31) U/L. The serum LAP activities of the two pregnant groups were much higher than that of control group (P

AIM: To observe the effect of partial liquid ventilation with perfluorocarbon on gas exchange, hemodynamics and lung histological change in the piglets with surfactant depletion-induced acute lung injury. METHODS: 12 piglets were treated by lung lavage to a partial pressure of oxygen in arterial blood (PaO 2) below 100 mmHg for one hour and randomly divided into gas ventilation group (control group) and partial liquid ventilation (PLV) group, in PLV group, piglets received PFC (FC3280) intratracheally at doses of 15 mL/kg. The parameters of gas exchange and hemodynamics were measured before lung lavage, after lung lavage when the acute lung injury (ALI) was established, and 1 hour, 2 hours after ALI. Animals underwent euthanasia at the end of the study, lung histologic analysis followed. RESULTS: Surfactant depletion by lung lavage induced a stable acute lung injury.Gas exchange increased markedly in the animals that underwent PLV, less hemodynamic damage was observed in PLV group compared with the animals in GV group. Lung histologic analysis demonstrated a less lung damage, including atelectasis, neutrophil excudation, intra-alveolar hemorrhage and interstitial edema in PLV group compared with control group. CONCLUSION: In piglets with surfactant depletion-induced acute lung injury, partial liquid ventilation with perfluorocarbon can improve the gas exchange with less adverse hemodynamic effect and less lung injury compared with conventional gas ventilation.

This study was intended to establish a method of purification of HPV16 L1 protein expressed in a prokaryotic system and to obtain the purified protein. The prokaryotic expression vector pGEX-4T-1-HPV16 L1 was constructed and transformed into E. coli BL21 cell, and induced by 1 mM IPTG to express HPV16L1 protein. The inclusion bodies were isolated and solubilized with 8 M urea. After the urea was removed by gradual dialysis, the denatured L1 protein were renatured and then were purified by affinity chromatography. The results showed that HPV16L1 protein formed inclusion bodies in bacterial expression system, suggesting that this assay can be used to purify HPV16L1 protein and hence provide a basis for studying the applications of HPV16 L1 protein.
Capsid Proteins ; biosynthesis ; Escherichia coli ; metabolism ; Genetic Vectors ; Human papillomavirus 16 ; Oncogene Proteins, Viral ; biosynthesis

ObjectiveTo assess the effects of constraint-induced movement therapy(CIMT) on sub-acute stroke patients with upper extremity motor dysfunction.Methods63 patients with hemiplegia after stroke were divided into two groups: control group(31 cases) and CIMT group(32 casese). The two groups received the regular rehabilitation training for 14 d. Then the control group went on the the regular rehabilitation training, while the CIMT group received CIMT for 14 d. All patients were assessed by the Action Research Arm Test(ARAT) and Fugl-Meyer(FMA) on the first day, the 15th day and the 30th day after the treatment.ResultsARAT score and FMA scores were higher in CIMT group than in the control group 30 d after the treatment and in CIMT group 15 days after the treatment(P<0.001).ConclusionCIMT is more effective to improve the upper limb motor function of the sub-acute stroke patients than the regular rehabilitation training.