Objective To construct short-hairpin RNA (shRNA) eukaryotic express vectors targeting of insulin like growth factor-1 receptor (IGF1R) gene, and to explore the changes of adhesion, invasion and FAK protein expression of MHCC-97 H hepatocellular carcinoma cells with RNA interference. Methods The shRNA oligonucleotide fragments were designed and synthesized based on the sequence of IGF1R mRNA. Double strands were formed after annealing and inserted into pGCsi-U6-Neo-GFP vector. The recombinant was transformed into Stable 3, then plasmids were extracted and identified by restriction enzyme and sequencing analysis. The most effective vectors were selected by RT-PCR and Western blotting after transfecting 293T cells. The best one was used to transform MHCC-97H cells which were selected with G418 to obtain positive colons. The changes of adhesion, invasion and FAK protein expression in MHCC-97H cells were studied. Results The restriction enzyme analysis and sequencing analysis demonstrated that shRNA had been inserted into vectors, and their sequences were the same as the design. It was indicated by RT-PCR and Western blotting that the silencing efficacy of IGF1R was 88%. The ability of adhesion and invasion significantly decreased after IGF1R silencing in MHCC-97H cells, and so was the expression of FAK protein. Conclusion IGF1R pGCsi-U6-Neo-GFP shRNA can significantly decrease the abilities of adhesion and invasion in MHCC-97H cells, and inhibit the expression of FAK protein.

Objective To investigate the accuracy of contrast enhanced ultrasound (CEUS) in diagnosing acute severe panereatitis.Methods Thirty three patients with acute severe pancreatitis were enrolled in the study and all of them underwent CEUS and spiral computed tomography(CT) scan.CT was taken as gold standard in acute severe pancreatitis diagnosis.CEUS were done within 72 hours after CT scan.Results Compared with CT, the sensitivity, specificity,accuracy, positive predictive value,and negative predictive value of CEUS for detecting the necrosis of pancreas were 90%, 95%, 94%, 90% and 95%respectively.The sensitivity, specificity, accuracy, positive predictive value,and negative predictive value of CEUS in diagnosing severe acute pancreatitis were 85%, 92%, 88%, 94% and 80% respectively, while 37% ,86% ,58%, 78%, 50% for conventional ultrasound respectively.Conclusions CEUS is accurate in detection of pancreatic necrosis and diagnosis of acute severe pancreatitis.

Objective To evaluate the clinical safety and efficacy of HIFU treatment of hepatic metastases from colorectal cancer.Methods 20 tumor lesions in 14 patients with hepatic metastases from colonectal cancer were treated with HIFU therapy.The vital signs,main organs'functions,examination of MRI and complications were observed.Results Vital signs of all the 14 patients were stable.The function of liver and kidney were normal and life quality were improved with elevation of Karnofsky in 5 patients.On MRI examination,the MRI signals of treating area of 20 tumor lesion in 14 patients decreased remarkably with disappearance of blood supply and a thin layer of edematous intensified zone was detected on the margin of lesion.6 patients were found to have cord-like burning scar on skin.Conclusion HIFU treatment is safe and effective in treatment of hepatic metastases from colorectal cancer.

Purpose:To study the activity of toremifene and its synergistic effect with anti-tumor drugs on human lung adenocarcinoma cell line A549,which might be expected to provide a new mode of therapy for the clinical management of lung cancer.Methods:The cytotoxic effects of these agents on human lung cancer cell line A549 were measured by a tet- razolium-based volorimetric assay (MTT assay).Results:Toremifene can inhibit the growth of A549 cell directly.The A value of the low dose toremifene combined with anti-tumor drugs were lower than that of anti-tumor drugs alone.Toremifene combined with VCR,ADM,DDP and VP-16 showed better effects.Conclusions:Toremifene combined with chemotherapy drugs shows significant synergistic anti-tumor effect on A549 cell.This might provide experimental evidence for lung cancer therapy.

To prepare thymosin alpha-1 (Talpha) loaded injectable sustained release microspheres and to evaluate its release behavior, bioactivities in vitro as well as its pharmacodynamics in vivo, Talpha1 loaded microspheres was prepared with poly ( lactic-co-glycolic acid) (PLGA) as carrier material by double emulsion (W/O/W) method. Physical and chemical properties of microspheres, such as mean diameter, The release behavior and its influencing factors were morphology and drug loading were evaluated. evaluated by HPLC determination. The bioactivity of Talpha1 in the course of encapsulation process and in vitro release ware evaluated by CCK-8 method. The ratio of CD4+/CD8+ in blood was determined with flow cytometry and the pharmacodynamics of Ta, loaded microspheres was evaluated by the change of CD4+/ CD8+. Microspheres with good shape and dispersive quality were prepared. The drug entrapment efficiency of two optimizing prescriptions containing 5% NaCl and 10% glucose as outer water phase were 87. 8% and 90. 2% , respectively. The cumulated release in one month is up to 90%. The bioactivity of Talpha was conserved with glucose as outer water phase, but in the course of in vitro release, the specific activity of Talpha in the microspheres decreased a little. Talpha microspheres can increase significantly the immunity of immuno-suppressed rats. Talpha can be encapsulated in injectable microspheres to yield one-month continuous release when using biodegradable polymers PLGA as carrier material, and this technique will have a favorable perspective in the near future.
Animals ; Biological Availability ; CD4-CD8 Ratio ; Delayed-Action Preparations ; Drug Carriers ; Drug Compounding ; methods ; Injections ; Lactic Acid ; chemistry ; Male ; Mice ; Mice, Inbred BALB C ; Microspheres ; Particle Size ; Polyglycolic Acid ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Thymosin ; administration & dosage ; analogs & derivatives ; chemistry ; pharmacokinetics

Child ; Humans ; Hypereosinophilic Syndrome ; drug therapy ; pathology ; Male ; Nephrotic Syndrome ; pathology

Female ; Humans ; Infant ; Male ; Nephrotic Syndrome ; congenital ; pathology ; therapy

Animals ; Antioxidants ; metabolism ; Fatigue ; blood ; Female ; Hypericum ; chemistry ; Male ; Mice ; Mice, Inbred ICR ; Physical Conditioning, Animal ; Plant Extracts ; pharmacology

Background and purpose: With the development of the means of preoperative diagnosis and treatment of breast cancer, ifne needle puncture no longer seems to be popular with the clinical use. The safety and effectiveness of fine needle aspiration as preoperative diagnosis of breast cancer are controversial. This study was aimed to investigate the application of fine needle aspiration (FNA) of breast cancer and association between FNA and recurrence and metastasis for 30 years in our hospital. Methods:A total number of 1 260 patients with breast cancer treated in Peking Union Medical College Hospital from 1975 to 2006 were reviewed in two groups for clinical characters, recurrence, and survival rate. The two groups were FNA and non-FNA. Results:The positive rate of FNA was 61.4%and the highest rate was inⅣstage (100%). StageⅡandⅢwere the major ones in FNA group (45.2%). The local recurrence (7.0%vs 6.4%), recurrence and metastasis (13.2%vs 14.0%), 5-year (82.0%vs 81.0%) and 10-year disease free survival rates (51.0%vs 64.0%) have no signiifcant difference between FNA and non-FNA groups. Conclusion:FNA is still one of the most important methods for preoperative investigation of breast cancer. Background and purpose: With the development of the means of preoperative diagnosis and treatment of breast cancer, ifne needle puncture no longer seems to be popular with the clinical use. The safety and effectiveness of fine needle aspiration as preoperative diagnosis of breast cancer are controversial. This study was aimed to investigate the application of fine needle aspiration (FNA) of breast cancer and association between FNA and recurrence and metastasis for 30 years in our hospital. Methods:A total number of 1 260 patients with breast cancer treated in Peking Union Medical College Hospital from 1975 to 2006 were reviewed in two groups for clinical characters, recurrence, and survival rate. The two groups were FNA and non-FNA. Results:The positive rate of FNA was 61.4%and the highest rate was inⅣstage (100%). StageⅡandⅢwere the major ones in FNA group (45.2%). The local recurrence (7.0%vs 6.4%), recurrence and metastasis (13.2%vs 14.0%), 5-year (82.0%vs 81.0%) and 10-year disease free survival rates (51.0%vs 64.0%) have no signiifcant difference between FNA and non-FNA groups. Conclusion:FNA is still one of the most important methods for preoperative investigation of breast cancer.

BACKGROUND:Nowadays, growth factors are commonly used to induce bone marrow mesenchymal stem cells. However, this is a high-cost method with a great amount of growth factors. In addition, the chondrogenic potential of bone marrow mesenchymal stem cells wil decrease significantly with increasing times of culture. OBJECTIVE:To observe the directed differentiation of bone marrow mesenchymal stem cells co-cultured with human synovial fluid. METHODS:Human bone marrow mesenchymal stem cells were isolated and cultured by adherence screening method. The synovial fluid of the knee was aspirated from healthy volunteers by aseptic operation. Passage 3 human bone marrow mesenchymal stem cells were co-cultured with the fol owing media:synovial fluid+complete medium;synovial fluid+bone marrow mesenchymal stem cells+complete medium;bone marrow mesenchymal stem cells+complete medium. The morphology and growth of the cells were observed under an inverted microscope every day. At days 7, 14 and 21 of induction, toluidine blue staining and immunocytochemical staining were performed.
RESULTS AND CONCLUSION:After co-culture with human synovial fluid, human bone marrow mesenchymal stem cells proliferated slowly, and varied from fusiform to oval or polygonal;toluidine blue and col agen II staining were positive. These findings indicate that the synovial fluid has a positive role in the chondrogenic differentiation of bone marrow mesenchymal stem cells. The synovial fluid may contain substances that promote the chondrogenic differentiation of bone marrow mesenchymal stem cells.