Objective To study the technique of transposon mutagenesis involving the use of Mini-Tn5 on a suicide plasmid for Citrobacter freundii (C. freundii). Methods A conjugation method was used in transposon mutagenesis. The stability of exconjugants was confirmed by series passages under nonselective condition. Detection of anti-ampicillin resistance and purification of the plasmid were conducted for excluding the possibility of plasmid-encoded antibiotic resistance. All exconjugants were screened for auxotrophic mutants and swarming motility mutants. Results Analysis of the generated exconjugants indicated that Mini-Tn5 Km was transposed in C. freundii at a high frequency (ca. 9.2?10 -6). The mutants were stable and retained the transposon-encoded antibiotic resistance after incubation for three passages under nonselective condition. The frequency of auxotrophic mutants in the population confirmed that the insertion of the transposon was random and the C. freundii chromosome did not contain significant insertional hot spots of transposition. No extrachromosomal plasmid DNA was found. Conclusion Transposon mutagenesis of Mini-Tn5 is a suitable technique for the study of differentiation in C. freundii.

CCK receptor belongs to G-protein-coupled receptor (GPCR) superfamily.Polymorphism of CCK receptors can alter drug affinity and/or biological efficacy, and its genetic differences in amino acid sequences can induce ligand-independent signaling, which in turn can lead to disease. With growing efforts in the field of pharmacogenomics, it is anticipated that polymorphism-induced alterations in drug and/or receptor function will be a focus of increasing concern in the future drug-development project. Study of CCK receptor polymorphism may reveal some universal rules in GPCR superfamily. In this review, the alterations of receptor function and/or drug efficacy resulted from polymorphism in CCK receptors will be discussed in the viewpoint of molecular biology and pharmacogenomics, and some strategies in development of receptor-specific drugs will be put forward.

OBJECTIVE: To probe into the application of antibiotics in orthopedic perioperative period. METHODS: The application of 591 patients with orthopedic disease in perioperative period from Jun. 2006 to Jun. 2008 were reviewed and analyzed. RESULTS: 591 patients were treated with antibiotics in perioperative period. 85 cases of preoperational medication accounted for 14% and 286 cases of drug combination 48%. Among 286 cases of drug combination, 263 cases of two-drug consisted of 92%, 23 cases of three-drug 8%; mean administration time was(9?2.3)d. CONCLUSION: Abuse, inappropriate use and inappropriate drug combination of antibiotics were existed, which need to be corrected.

With advances in arthroscopic surgery, arthroscopic repair for rotator cuff tears is often used by surgeons. Surgi?cal methods have already developed from simple single?row fixation and double?row fixation to complicated suture?bridge fixation. Although arthroscopic suture?bridge rotator cuff repair has been proposed to be advantageous over single?row and double?row fixa?tion in biomechanical constructs, the failure of rotator cuff repair is still higher. Different suture methods of medical row anchor in arthroscopic suture?bridge technique may change the suture?tendon loading and decrease the retear of suture?tendon. And differ?ent fixation methods of lateral row anchor in arthroscopic suture?bridge technique may decrease the anchor pull?out. These will be useful to decrease the failure of arthroscopic suture?bridge technique. There will be a good outcome if optical suture and fixation approach can be chosen according to the quality and retraction of rotator cuff, suture tension and the bone quality of the greater tu?berosity in arthroscopic suture?bridge technique.

AIM: To investigate the efficacy of intravitreal ranibizumab combined with laser therapy in the treatment of macular edema caused by branch retinal vein occlusion ( BRVO) .
METHODS:There were 78 patients (78 eyes) who were diagnosed with macular edema caused by BRVO using fundus fluorescence angiography ( FFA ) and optical coherence tomography ( OCT ) . Group A: randomly selected 26 cases ( 26 eyes ) were given grid laser photocoagulation ( GLP) . Group B: randomly selected 26 cases (26 eyes) were given GLP first, and then received intravitreal ranibizumab 1wk later. Group C: randomly selected 26 cases ( 26 eyes ) undergone intravitreal ranibizumab first, and then given GLP 1wk later. There was no significant difference in macular edema. We analyzed the changes in the best corrected visual activity ( BCVA ) , central macular thickness ( CMT ) before and 1wk, 1,6mo after treatment.
RESULTS: Compared with before treatment, 1wk after treatment: mean value changes of BCVA and CMT were no significant difference in group A (P>0. 05);mean value changes of BCVA was improved and mean value of CMT was decreased in groups B and C, the difference was statistically significant (P<0. 05);BCVA and CMT changes between groups had statistical significance ( P<0. 01 );After treatment 1, 6mo:compared with before treatment, mean BCVA was improved in three groups, mean CMT was lower, the differences were statistically significant (P<0. 01);BCVA and CMT changes between groups were statistically significant ( P<0. 05 ); compared with 1mo after treatment, mean BCVA and CMT were no significant difference (P>0.05) between groups A and B after 6mo treatment;mean BCVA improved and CMT average value was decreased in group C, the difference was statistically significant ( P<0.01 ); BCVA and CMT changes between groups were statistically significant (P<0. 01).
CONCLUSION: Intravitreal injection ranibizumab combined laser therapy can effectively reduce BRVO induced macular edema, enhance vision acuity. Compared with GLP, combination therapy has more rapid onset of treatment, and reduce macular edema better; Intravitreal ranibizumab should be given in front of the GLP, and the treatment effect is more precise, more stability.

Adolescent ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Blood Pressure ; drug effects ; Child ; Child, Preschool ; Female ; Fosinopril ; pharmacology ; therapeutic use ; Humans ; Male ; Nephritis ; drug therapy ; Proteinuria ; drug therapy ; Purpura, Schoenlein-Henoch ; complications

Objective To study the changes of system immune and intestinal immune in the progression of non-alcoholic fatty liver disease due to obesity. Methods Ninty male SD rats were divided into control, high-sucrose and high-fat diet groups. Non-alcoholic fatty liver disease models were established by feeding with high-sucrose diet or high-fat diet and were killed at the 4th,8th and 12th weeks with 10 each for each group. The extent of liver steatosis was observed with HE staining.Portal blood endotoxin level was assessed by limulus test. The percentage of CD4+ and CD8+ cells in peripheral blood mononuclear cells (PBMC) and lymphocytes in Peyer's patches (PP) were calculated by flowcytometry. Results In comparison with control group, the endotoxin level was not elevated from the 4th week to 12th week in high-sucrose diet group, (all P values＞0.05), but was increased in high-fat diet group at the 8th week (P＜0.05). CD4/CD8 ratio in PBMC was higher in high-sucrose and higt-fat diet groups than that in control group at the 4th week (P＜0. 05) ,but was lower than that in control group at the 8th and 12th weeks (P＜0. 05). Whereas the variation of CD4/CD8 ratio in PP was consisted with that in PBMC between the high-sucrose and high-fat diet groups at the 4 th and 8 th weeks, but there was no difference when compared with control group at the 12 th week (P＞0.05).Conclusion Obesity can inhibit systematic immune and intestinal immune. The intestinal immune may be regulated by the liver.

Objective To investigate the clinical autopsy results of patients died of cardiovascular disease or other disease complicated with cardiac damage. Methods Complete autopsy was performed on 86 cases with uncertain cause of death. Through integrating clinical diagnosis and treatment with gross autopsy findings and microscopic observations, 86 autopsies were determined the major cause of death. Results In 86 autopsies, 69 cases were heart disease. Differences between pathological diagnosis and clinical diagnosis were compared. Twenty-seven cases were cardiac deaths, with diagnosis accrodance rate of 81. 5%. Fortytwo cases died of non-cardiac disease but complicated with heart disease or involving the heart which accelerated the death in patients, with accordance rate of 78.6%. Conclusion Scientific and correct performance of autopsy was important to determine the causes of death, to promote development of related disciplines and to improve the diagnosis and treatment of the diseases.

Objective To select the optimum condition for the determination of formaldehyde in the car and to understand the formaldehyde pollution situation,dynamic change law and the influence factors.Methods The national standard method,acetyl acetone spectrophotometer,was used to determine formaldehyde.The investigated cars were the ones which left the factory for one month,three months,six months and one year.The sampling time,rate of flow and coloration time were determined.The influence of temperature and ventilation on the formaldehyde content in the car was analyzed.Results The formaldehyde content in the car was influenced by the temperature,ventilation,date of production and the type of the car.The optimum conditions for the determination of formaldehyde in the car were as follows,10 min of sampling time,0.3 L/min of sampling flow rate and 3 min of coloration time.Conclusion Ventilation is an effective measure to reduce the formaldehyde content in the car.Higher temperature is good for the release of formaldehyde.The formaldehyde content in the car will reach the standard limit in one year after left factory.

Aim To detect the effect of a combination of extracts of ginseng and ginkgo biloba(Naoweikang,NWK) on acetylcholine,monoamines and their metabolites in ?-amyloid peptide(A?)_(1-40) treated rats and the potential mechanisms.Methods A 1-month NWK(15.5,31 and 62 mg?kg~(-1),respectively) administration to rats was performed daily after bilateral injection of A?_(1-40)(4 g?L~(-1) for each side) into hippocampus.Acetylcholine(ACh) was determined with an improved HPLC-ECD method,which is combined with two immobilized enzyme reactors.Monoamines and their metabolites were also determined with HPLC-ECD.Result Compared with shams,ACh and serotonin(5-HT) in whole brains in models decreased significantly(P