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.

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.

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.

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

OBJECTIVETo study the correlation between intrusion of nucleus gelatinosus into vertebral body and low back pain after thoracolumbar fractures.

METHODSOne hundred and two patients who were treated in Peking university third hospital from January 2005 to September 2007 were evaluated retrospectively and included into this study. They were followed up for an average of 18 months and complete data were kept. Fifty-four patients were treated operatively and forty-eight patients were treated conservatively. VAS questionnaire was applied to evaluate the low back pain. Spearman Correlation Coefficients module and Wilcoxon 2-Sample Test module of SAS software were used to analyze the relation between intrusion of nucleus gelatinosus, different treatments, kyphotic Cobb angles and VAS.

RESULTSThere were strong correlations between intrusion of nucleus gelatinosus and VAS in both operative group, P < 0.01 and non-operative group, P < 0.01; there were poor correlations between different treatments and VAS in both intrusion group, P > 0.05 and non-intrusion group, P > 0.05; and there were also poor correlations between Cobb angles and VAS, gamma = 0.2584, P > 0.05.

CONCLUSIONThere is correlation between the intrusion of nucleus gelatinosus into vertebral body and the low back pain after thoracolumbar fractures.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Back Pain ; etiology ; Female ; Humans ; Intervertebral Disc ; pathology ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Spinal Fractures ; complications ; pathology ; Thoracic Vertebrae ; injuries ; Young Adult

The index components contents of different time and different stubbles in honeysuckle were measured by HPLC, and were analysis by using the method of SPSS. Results showed that the content of index ingredients of different time had differences, and firstly decreased, then increased with time. The content of index ingredients of different stubbles had significantly differences, and firstly decreased, then increased with time. The chlorogenic acid contents were 2.059%-3.593%. The luteolosid contents were 0.110%-0.171%. Results indicated that the best picking buds time is before seven o'clock in the morning and evening at before and after seven o'clock, the index component content is higher. Picking buds in spring and at autumn index component content is higher; Picking buds in summer index component content is low. The experiment provides theoretical support for quality control in the whole process of the honeysuckle harvested and comprehensive utilization of honeysuckle.
Chlorogenic Acid ; analysis ; Drugs, Chinese Herbal ; analysis ; Flowers ; chemistry ; growth & development ; Lonicera ; chemistry ; growth & development ; Luteolin ; analysis ; Seasons ; Time Factors

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

To study the effect of miR-490 on Coxsackievirus B3 (CVB3) replication, HeLa cells were trans- fected with miR-490 in vitro, and infected with a Renilla luciferase (RLuc)-expressing CVB3 variant (RLuc-CVB3). The activities of RLuc in these cells were measured at 8h intervals from 0 to 40 h post-infection (p.i.), and the effects of miR-490 on RLuc-CVB3 replication were observed. In a further study, HeLa cells were transfected with either miR-490 or antisense miR-490 (AMO-miR-490), and were then infected with an enhanced green fluorescence protein (EGFP)-expressing CVB3 variant (EGFP-CVB3). The replication of EGFP-CVB3 was then determined by detecting the expression of EGFP. We observed that miR-490 could significantly inhibit the expression of RLuc in infected cells at 32 h p. i. Furthermore, in HeLa cells infected with EGFP-CVB3 at 32 h p.i., EGFP expression was also significantly inhibited by the presence of mniR-490. The inhibitory effect of miR-490 on EGFP expression in EGFP-CVB3-infected cells could be reversed by tranfection with AMQ-miR-490. These results indicated that miR-490 significantly inhibits the replication and expression of QVB3.
Enterovirus B, Human ; genetics ; physiology ; Enterovirus Infections ; genetics ; metabolism ; virology ; HeLa Cells ; Humans ; MicroRNAs ; genetics ; metabolism ; Virus Replication