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

Multilocus sequence typing (MLST) is a molecular genotyping method based on nucleotide sequencing. The procedure of this method characterizes isolates of bacterial species using the DNA sequencing of multiple housekeeping genes(usually seven). For each housekeeping gene, the different sequences present within a bacterial species are assigned as distinct alleles.For each isolate, the alleles at each of the loci define the allelic profile or sequence type (ST). MLST has the advantages of being robust (based on genetic data) and electronically portable to generate data that allow rapid and global comparisons between different laboratories. In this paper, the principle, method, data analysis, application, advantages and flaws of MLST are introduced.

Objective: To explore the expression level of serum interleukin (IL)-7 in patients with acute coronary syndrome (ACS) and analyze the relationship between IL-7 level and prognosis. Methods: A total of 130 ACS patients [ACS group, including 70 cases with acute myocardial infarction (AMI) and 60 cases with unstable angina pectoris (UAP)], 33 cases with stable angina pectoris (SAP，SAP group) and 89 healthy subjects (healthy control group) were selected. IL-7 level was measured using enzyme linked immunosorbent assay (ELISA) and compared among all groups. The 130 ACS patients were followed up, and Logistic regression analysis was used to analyze the relationship between IL-7 level and prognosis. Results: Compared with healthy control group and SAP group, there was significant rise in IL-7 level in UAP group and AMI group [(1.84±0.47) pg/ml, (2.11±0.63) pg/ml vs. (4.87±0.52) pg/ml, (5.15±0.71) pg/ml, P<0.05 or P<0.01]. There were no significant difference in IL-7 level between healthy control group and SAP group, UAP group and AMI group (P>0.05 both); Logistic regression analysis indicated that expression level of serum IL-7 was an independent risk factor for adverse cardiovascular events in ACS patients (OR=1.212, 95%CI:1.061-1.418). Conclusion: Interleukin-7, as an important inflammatory cytokines, its serum level abnormally elevated in patients with acute coronary syndrome, it may have important prognostic value.

Objective To investigate the molecular epidemiological characteristics of Staphylococcus aureus associated with bloodstream infection in hospital. Methods 47 Staphylococcus aureus strains isolated from bloodstream in PLA General Hospital were collected from January 2006 to December 2008. Susceptibility of the strains to 11 antimicrobial agents was detected and DNA homology of them was analyzed with Rep-based DiversiLab~(TM) Microbial Typing System. Panton-Valentine leukocidin (PVL)gene was determined by PCR. For methicillin-resistant Staphylococcus aureus (MRSA) strains,the genotypes of SCCmec were determined and ST239 clone was screened with multiplex PCR. Multilocus sequence typing (MLST) was used to determine the STs of the selected isolates. Results In the 47 Staphylococcus aureus isolated from blood,methicillin-resistant strains accounted for 51.1%,all belonged to SCCmec Ⅲ type,with only 2 pvl gene positive strains identified. 12 different patterns (A-L) were found among 47 strains with Rep-PCR. All MRSA strains clustered in the A and B subtypes. Conclusion Most MRSA strains isolated from blood in PLA General Hospital belonged to ST239-MRSA-SCCmec Ⅲ clone. DiversiLab~(TM) Microbial Typing System could provide a rapid and effective method to investigate the molecular epidemiological characteristics of Staphylococcus aureus in the hospital settings.

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

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