OBJECTIVE Apolyclonalantibody-basedhomogeneouschemiluminescenceimmunoas-say was developed and optimized using AlphaLISA technology for the quantitative detection of microcys-tin-LR(MC-LR)inwatersamples.METHODS Thismethodwasbasedonacompetitivemodelin which an immune complex was formed from the ingegral binding of artificial MC-LR antigen-coated lumi-nescene beads,free MC-LR standards or sa mples,antibody and biotinylated second antibody.Next sensor bead were added that approached the i mmune co mplex through biotin-streptavidin interaction. With the exciting light,the energy was passed from the sensor luminescer before a special emission light could be observed.To opti mize the reaction conditions,working dilutions of polyclonal antibody and bioti-nylated second antibody were assayed while the effect of buffer syste ms and ti me of each reaction were evaluated.RESULTS Maininfluencingfactorsoftheassaywerediscussedasworkingdilutionsofpoly-clonal antibody and biotinylated goat anti rabbit IgG,assay buffer and reacting ti me.After opti mization of reaction conditions,MC-LR AlphaLISA could be finished in 40 min,with a sensitivity of 0.006 μg·L-1 and a dynamic range of 0.006 -5 μg·L-1 .The coefficient of variation was below 10% and average recovery was 1 07.7%.Moreover,the cross reactivity rates of MC-RR and MC-RY to MC-LR were 13.2%and0.91%,respectively.CONCLUSION Thismethodishighlysensitiveandspecific,time-saving and quite suitable for high throughput determination of MC-LR water samples.

Objective To explore the histopathological manifestations of various complications after transplantation,the patients in our hospital after liver transplantation liver puncturebiopsy pathology data were analyzed,and then provide a reliable clinical diagnosis and treatment for the patients scheme.Methods A retrospective analysis of our hospital 198 cases of liver transplantation in 249 cases diagnosis of liver puncture biopsy pathology data,HE staining method to analyze the pathological morphological changes,using rejectionpathological criteria,according to clinical examination and treatment effect of international unified Banff.Results All biopsy materials,acute rejection rate is the highest,a total of 71 cases(28.5％),biliary complications occurred in 39 cases (15.7％),hepatitis B virusinfection and recurrence in 28 cases (11.2％),34 cases of drug-induceddamage (13.7％),reperfusion injury in 35 cases (14.1％),CMV infection of 14cases (5.6％),tumor recurrence in 7 cases (2.8％),chronic rejection in 16cases (6.4％),primary graft non function in 2 cases (0.8％),it is difficult to determine in 3 cases (1.2％).Conclusion Transplantation of liver biopsy can provide correct cause for abnormal liver function,and to guide the clinical treatment of accurate,effective treatment,suggested that thetransplantation center will transhepatic listed for liver transplantationpostoperative routine inspections,periodic biopsies,the survival of the state to better protect the graft.

Aim To develop a new method for easy endotrache-al intubation and instillation in mice. Methods Relying on the endoscope of cold light and gel loading tips, a new method was set up for intubated operations. Under high magnification, oper-ations of endotracheal intubation and instillation were completed in mice. Twenty male CB17-SCID mice ( between 24 and 28 g) were repeatedly intubated and instilled with 80 μL suspensions of air polluted fine particulate matters in a week. Results The twenty mice were sacrificed after 24h of the last instillation. The dispersions of the air polluted fine particulate matters were found at the bottoms of tracheas and in both the left and right lungs in all mice. Conclusion Using the endoscope of cold light and gel loading tips, a new method for tracheal instillation is set up in mice, which is rapid, simple and has good repeatability. It could be widely applied for the endotracheal intubation and in-stillation in mice.

Objective To observe the clinical curative effect of drug combined with psychological treatment on patients with sleep disorders after stroke.Methods A total of 180 cases of eligible patients with sleep disorders were included and randomly di-vided into control group and treatment group.Control group were treated with conventional vascular drug(estazolam,2 mg,before sleep);treatment group increased anxiolytic and sedative hypnotic drugs(venlafaxine,dexzopiclone)compared to control group.Af-ter 14 days,sleep quality,NIHSS were compared both within the treatment group and between the two groups.Results The Bar-thel index,NIHSS and PSQI of two groups had significant differences,and those index of treatment group before and after treat-ment had significant differences too(P <0.05).Conclusion Combined conventional cerebrovascular drugs with Venlafaxine,dexzo-piclone as well as psychological treatment significantly improved sleep quality of patients with acute stroke.

OBJECTIVETo observe the surface of Enterococcus faecalis and the dynamic forming process of those biofilms using atomic force microscopy (AFM) in air condition.

METHODSThe surface of Enterococcus faecalis which were dried in air were observed with AFM. We used the cellulose nitrate film to construct the Enterococcus faecalis biofilms model in vitro, and then placed the biofilms under AFM to observe the surface changes of biofilms' development.

RESULTSThe cell surfaces of strain Enterococcus faecalis were not regular because of the presence of the amorphous substance on the colony surface, which congregated globular, fibrous structure. Gradually determined that at 6 h the initial biofilm formed and at 24 h the biofilms maintained the steady-state. AFM height images showed topographical changes due to biofilms' development, which were used to characterize several aspects of the bacterial surface, such as the presence of extracellular polymeric substance, and the biofilms' development stage.

CONCLUSIONApplication of AFM in physiological conditions could be useful in observing Enterococcus faecalis surface ultrastructure and dynamic process of biofilms formation.

Bacterial Adhesion ; Biofilms ; Enterococcus faecalis ; growth & development ; Microscopy, Atomic Force

Objective To investigate use of magnetic resonance diffusion tensor imaging (MR-DTI) for quantitatively evaluating the efficacy of Mailuoning Injection on non-compressive lumbar radiculitis.Methods Nine Bama mini pigs were selected and divided into group A,B and C,3 cases in each group.The non-compressive lumbar radiculitis model was established under CT-guiding.The corresponding therapeutic drugs (group A:10 mL Mailuoning;group B:10 mL normal saline;group C:10 mL diminishing inflammation fluid) were given by epidural injection on 14 d after constructing model.MR-DTI was performed before model construction,14 d after model construction and before treatment.One experimental pig in each group was taken on 3,7,14 d after treatment,performed MR-DTI and killed for taking the nerve root sample to conduct the immunohistochemical detection.The fractional anisotropy (FA) values of nerve root in MR-DTI imaging were measured.The FA values and immunohistochemical detection results were statistically analyzed.Results MR-DTI:the FA values after model construction in each group was decreased (P<0.05);the FA values on 14 d after treatment in the group A was increased,which showed statistically significant difference compared with before treatment (P<0.05);the FA values after 3,7 d treatment in the group A had no obvious increase,the difference between the group A and B was not statistically significant (P>0.05),but the FA values increase in the group C was earlier and more rapid than other two groups (P<0.05);the FA values after 14 d treatment in the group A was risen again,the FA values of bilateral nerve roots had no statistical difference between the group A with the group B and C (P>0.05),while the FA values had statistical difference between the group B and C (P<0.05).The immunohistochemical results:TNF-α integral absorbency value(IA value) on 7 d after treatment in the group A began to decline;the TNF-α IA value on 14 d after treatment in the group A and C was significantly decreased compared to group B,the difference was statistically significant (P<0.05).Conclusion Mailuoning Injection has a certain effect on non-compressive lumbar radiculitis,which can be evaluated by using DTI.

Adolescent ; Adult ; Anxiety ; epidemiology ; Family Relations ; Female ; Humans ; Male ; Students, Medical ; psychology ; Young Adult

A 44-residue hybrid peptide (CB (1-24)-Arg-Ser-Tyr-Tan (4-21)) incorporating 1-24 residues of cecropin B (CB) and 4-21 residues of thanatin (Tan) was designed and constructed. The CB-Tan gene was cloned into expression plasmid pGEX-3X and expressed in E. coli BL21. The fusion protein was purified by affinity chromatography. After digested with enterokinase the gene product released with antibacterial activity and gave one band in Tricine-SDS-PAGE.
Amino Acid Sequence ; Anti-Bacterial Agents ; metabolism ; Antimicrobial Cationic Peptides ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; Insect Proteins ; biosynthesis ; Molecular Sequence Data ; Peptides, Cyclic ; biosynthesis ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification ; pharmacology

OBJECTIVETo investigate the antibacterial effect of a particular antimicrobial peptide Cecropin B(CB) on Pseudomonas aeruginosa infection of wound in mice.

METHODSThirty ICR mice were enrolled in the study, and the Pseudomonas aeruginosa infection model was reproduced by excision of the full layer of dorsal skin with an area of 1 cm x 1 cm. Then they were randomly divided into C ( control, n = 10, with wet compress of isotonic saline at 3 postinjury hour( PIH) ) , M (with hydropathic compress of 100 g/L mafenide at 3 PIH), A (with wet compress of 1 000 mg/L Cecropin B at 3 PIH) groups. The changes in body temperature and hemogram in each group were determined before and 4 days after injury. Quantitative examination were used to detect the quantity of bacteria in muscular tissue of the wounds, and the survival of the mice were observed on 4 post-injury day( PID).

RESULTSThe wounds were moist with more exudation in C group,while that in other groups were dry without obvious exudation. The body temperature of the majority of the mice in each group were elevated, but the number of leucocytes in each group was lowered after operation. The quantity of bacteria in muscle in A group[ (42 +/- 50) CFU/g] was obviously lower than that in M group [(886+/-804) CFU/g, P <0.05] , and it was all obviously lower than that in C group[ (41 +/-28) x 10(5) CFU/g, P <0.01]. The number of surviving mice after 4 PID in C group was evidently smaller than that in A and M groups( P <0. 05).

CONCLUSIONThe cecropin B possesses obvious anti-bacterial effect on the Pseudomonas Aeruginosa infected wounds of ICR mice, and it can reduce the mortality.

Animals ; Antimicrobial Cationic Peptides ; therapeutic use ; Disease Models, Animal ; Insect Proteins ; therapeutic use ; Male ; Mice ; Mice, Inbred ICR ; Pseudomonas Infections ; drug therapy ; Wound Infection ; drug therapy ; microbiology

OBJECTIVETo investigate the association between genetic polymorphisms of human leukocyte antigen-DQ (HLA-DQ) and susceptibility to trichloroethylene (TCE)-induced severe generalized dermatitis.

METHODSA case-control study was conducted which included 112 patients with TCE-induced severe generalized dermatitis and 142 healthy controls exposed to TCE in the same workshop. The DNA sequences in exon2 of HLA-DQA1 and HLA-DQB1 were performed by direct sequencing of polymerase chain reaction (PCR) products. The frequencies distribution of allelic genotypes and codon polymorphisms were compared.

RESULTSThe frequencies of DQA1*0201 and 060101/0602 in cases [7.6% (17/224) and 16.1% (36/224)] were significantly higher than those of the exposed controls [3.5% (10/284) and 7.0% (20/284)], while frequencies of DQA1*0103 and 050101/0503/0505 in cases [5.8% (13/224) and 8.9% (20/224)] were significantly lower than those of exposed controls [10.9% (31/284) and 17.3% (49/284)]. In terms of codon polymorphisms, there were 5 codons of DQA1 (25, 41, 52, 54 and 69) showing significant differences between cases and controls. There were no significant differences in the frequencies of allelic genotypes of HLA-DQB1 between cases and exposed controls.

CONCLUSIONThe genetic polymorphisms of HLA-DQA1 might be one of the factors influencing the individual susceptibility to TCE-induced severe generalized dermatitis.

Adolescent ; Adult ; Alleles ; Drug Eruptions ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; HLA-DQ Antigens ; genetics ; Humans ; Male ; Occupational Exposure ; Polymorphism, Genetic ; Trichloroethylene ; toxicity