Objective To evaluate the clin ical value of Multi-slice helical CT three dimensional angiography (3D-MSCTA) as first method for diagnosing intracr anial aneurysms. Methods We studied patients with clinical suspected intracranial aneurysms (13 patients with subarachnoid hemorrhage among cases). All these patients under went 3D-MSCTA and Digital Subtraction Angiography (DSA), 16 patients of them accepte d operation treatment. Row data was acquired by Multi-slice helical CT-AQUILION (Toshiba): scan speed 0.5 s/rot, image slice thickness 1.0 mm, helical pitch 3 .0/5.0. Contrast media (Angiografin) was injected intravenously (1.0-2.0 ml/kg) at spee d of 2.5-3.0 ml/s, delay time was 15-23 sec, reconstruction interval 0.5 mm, recons truction slice thickness 1.0 mm. Source images were processed using a workstation SGI-O2 , images post-processing software was ALATOVIEW,Version 1.21. The reconstructed images were then processed into shaded volume rendering (SVR) and maximal intensity projection (M IP) and Fly-through images. Entire brain DSA was performed obtaining anterioposteri or, lateral, and oblique images. Images of 3D-MSCTA and DSA were analysed by 3 radiologists and 2 neurosurgeons. Results 25 aneurysms were d etected by 3D-MSCTA. Aneurysms′s body, neck, source vessel and the relationship between the aneurysm and surrounding structures was clearly and surely displayed. 22 of 25 aneurysms were detected by DSA,another 3 were (1 anterior communicating artery aneurysm and 2 left middle cerebral artery aneurysm) was not detected. Sixteen of patients un derwent operation treatment, and the results of 3D-MSCTA corresponded very well to those of operation. Maximal diameter of aneurysms body was 14.0 mm and minimal diameter w as 1.7 mm. Conclusion 3D-MSCTA is a high sensitivity and rapid and no ninvasive method for detecting intracranial aneurysms. We suggest that 3D-MSCTA may be the first cho ice for diagnosing intracranial aneurysms.

Quantitative specific detection of Staphylococcus aureus is based on recombinant lysostaphin and ATP bioluminescence. To produce recombinant lysostaphin, the lysostaphin gene was chemically synthesized and inserted it into prokaryotic expression vector pQE30, and the resulting expression plasmid pQE30-Lys was transformed into E. coli M15 for expressing lysostaphin with IPTG induction. The recombinant protein was purified by Ni(2+)-NTA affinity chromatography. Staphylococcus aureus was detected by the recombinant lysostaphin with ATP bioluminescence, and plate count method. The results of the two methods were compared. The recombinant lysostaphin was successfully expressed, and a method of quantitative specific detection of S. aureus has been established, which showed a significant linear correlation with the colony counting. The detection method developed has good perspective to quantify S. aureus.
Adenosine Triphosphate ; chemistry ; Chromatography, Affinity ; Escherichia coli ; Luminescent Measurements ; methods ; Lysostaphin ; chemistry ; Recombinant Proteins ; chemistry ; Staphylococcus aureus ; isolation & purification

BACKGROUND:After peripheral nerve injury, to inhibit scar formation by drugs is the key to functional recovery. To reduce the amount of scar formation we designed a prednisolone-loaded film which can sustain drug release and good achievement in in vitro drug release test.
OBJECTIVE:To prepare the prednisolone implantable film and investigate its in vivo biocompatibility and safety.
METHODS:Prednisolone-loaded nanoparticles were first prepared with reverse micellar emulsion-solvent evaporation method, and the composite film and drug-loaded film were further prepared. Then, we investigated the in vivo biocompatibility of drug-loaded film through celltoxicity test, hemolysis test, acute systemic toxicity test, chronic systemic toxicity test.
RESULTS AND CONCLUSION:After cultured for 7 days, the relative growth rate of L929 mouse fibroblasts was 92.6%, showing no cytotoxicity. The hemolysis rate of the film was 0.59%, indicating that the material had no hemolysis action. No abnormal biological behaviors were seen in mice after intraperitoneal injection of film extracts, and there were no changes in liver and renal functions in rats. As il ustrated above, we can safely come to a conclusion that prednisolone-loaded film possesses good biocompatibility and can be safely used in the experiment of reducing the scar at sites of peripheral nerve repair.

Background Endophthalmitis is a serious complication of intraocular surgery.Conventional identification methods for bacteria are becterial culture and smear method,but these laboratory tests spend a long time and have low positive rates.16S rDNA is the bacterial chromosome encoding ribosomal RNA sequences,and it is determined that 16S rDNA sequencing has a high specificity for the identification of bacteria.Objective This study was to identify the infectious bacteria from aqueous humor or vitreous body in the eyes with endophthalmitis by 16S rDNA sequencing technique,and to investigate the diagnosis efficency of 16S rDNA sequencing technique on bacterial endophthalmitis.Methods Anterior chamber fluid (0.1-0.2 ml) or vitreous humor (0.5-1.0 ml) specimens were collected from 5 eyes of 5 patients with endophthalmitis in Qingdao Eye Hospital from June to December 2015 and used for high throughput sequencing,bacterial culture and smear,respectively.Bacteria DNA was extracted from the specimen with D3096-01 trace DNA kit for the amplification of V3-V4 region of 16S rRNA gene and sequencing of hypervariable region of all microbes in the samples by MiSeq Illumina Sequencing Platform.Then the bioinformatic analysis including analysis of taxonomy,abundance and alpha diversity were performed.Nucleasefree water of 50 μl in the centrifuge tube was used as control.Results Five aqueous humor or vitreous body samples were collected,and the positive results were exhibited by smear examination,with the Gram positive bacilli in the trumatic endophthalmitis eye and Gram negative bacilli in the filtering bleb infectious endophthalmitis eye,and all culture results were negative.16S rDNA squencing showed the positive outcomes in all the 5 samples.The high abundent nacteral genuses were Staphylococcus (65.28％),Streptococcus (18.90％) and Pseudomonas (12.76％) in the trumatic endophthalmitis eye;the major components of sample were Pseudomonas (53.68％),Acinetobacter (8.62％) and Limnobacter (5.96％) in the eye with acute endophthalmitis occurring at 2 days following cataract surgery;the major components in the filtering bleb infectious endophthalmitis eye were Moraxella (88.89％) and Pseudomonas (9.52％);the Pseudomonas was major components in the later-onset endophthalmitis eye (84.63％) and the eye with acute endophthalmitis occurring at 1 day after cataract surgery (97.89％).Conclusions A distinct advantage is found in 16S rDNA sequencing technique for the indentification of the pathogenic bacteria in endophthalmitis eyes due to its high positive rate in comparison with bacterial culture and smear method.

A simple rapid detection of antibody to hepatitis delta virus (anti-HDV) in human serum was developed by using double antigen sandwich ELISA. HDV gene fragment encoding HDAg was isolated from a Chinese patient infected with HDV by RT-PCR, and a high-efficient expression HD-PQE31 strain was constructed with the fragment. We obtained high titer and good quality hepatitis delta virus protein purified by Ni-NTA metal-affinity chromatography, which was identified by Western blot and ELISA, then we set up the double antigen sandwich ELISA for detection of anti-HDV in human serum, and the performance of the sandwich ELISA was evaluated in terms of specificity and sensitivity. Results were: 1) The purified HDAg protein's purity was 90%, and its ELISA titer was 1/100 000. 2) 42 anti-HDV positive sera were detected and showed that the sensitivity of sandwich ELISA was higher than that of competitive ELISA (t=2.44, p<0.01). 3) The inhibitory rates for 2 anti-HDV positive sera by the specific HDAg were 74% and 93% respectively. 4) For the assay of specificity, all 60 samples infected by other hepatitis viruses and 30 normal samples were negative for anti-HDV. These results suggested that the double antigen sandwich ELISA with purified recombinant HDAg showed higher specificity and sensitivity, It can be used in routine laboratories to diagnose the HDV infection.

Objective To investigate the effects of glutamate receptor signaling on melanoma cell dendrite morphology and cytoskeleton protein. Methods A metastatic human malignant melanoma cell line WM451LU was cultured and transfected by recombinant adenovirus vector carrying a cDNA encoding microtubule-associated protein 2a (MAP2a). MK-801, an antagonist of N-methyl-D-aspartate receptor (NMDAR), and CPCCOEt, an antagonist of metabotropie glutamate receptor 1 (mGluR1), were used to treat transfected or untrausfeeted WM451LU cells. Confocal microscopy and three dimensional atomic force microscopy were used to assess subcellular location of NMDAR2A, mGluR1 and MAP2a as well as the dis-tribution of α-tubulin in and dendrite morphology of WM451LU cells. The proliferation of WM451LU cells was estimated by cell survival growth curve. Results Confocal laser microscopy revealed that NMDAR2A, mGluRl and MAP2a were mainly co-localized in melanoma cell dendrites. Both MK-801 and CPCCOEt increased the density of microtubules in cell dendrites and dendritic branching of WM451LU cells, and both effects of MK-801 and CPCCOEt were enhanced by the expression of MAP2a. Furthermore, the proliferation of WM451LU cells was significantly inhibited by MK-801 of 100 μmol/L and CPCCOEt of 10 μmol/L. Conclusions In melanoma cells, glutamate receptors may participate in the development of dendrites, and anta- gonists of glutamate receptors could inhibit the proliferation of melanoma cells.

OBJECTlVE To study the anti-HBV activity of prepared recombinant human serum aIbu-min-interferon α-2b fusion protein(HSA-IFNα-2b) in vitro. METHODS HepG2 ceIIs were infected with recombinant adenovirus with green fIuorescence protein and 1.6-foId HBV DNA(AdGFP-HBV). The ex-pression of HBV antigens,HBsAg and HBeAg in cuIture medium was detected by ELISA assay. The tox-icity of HSA-IFNα-2b on HepG2 ceIIs was evaIuated by mTT assay.The reIative expression of HBV RNA in ceIIs and the absoIute quantity of HBV DNA in cuIture supernatant were determined by quantitative PCR assay. The activity of HBV enhancer Ⅰ was detected by DuaI-Reporter gene assay. RESULTS HBV couId repIicate and express in HepG2 ceIIs after infection with AdGFP-HBV. The expression of HBsAg and HBeAg in cuIture serum of HepG2 ceIIs infected with AdGFP-HBV decreased by 51.32%(P﹤0.01)and 50.26%(P﹤0.01),respectiveIy,when HSA-IFNα-2b 500 kU·L-1 was added. The same concentration of HSA-IFNα-2b didn't inhibit the proIiferation of HepG2 ceIIs,but inhibited HBsAg in a concentration-dependent manner. The regression formuIa between HBsAg inhibitory rate(Y)and con-centration of HSA-IFNα-2b(X)was Y=21.11 IgX+11.91(r 2 = 0.954),IC50 = 63.76 kU·L-1 . HBV RNA in ceIIs and HBV DNA in the cuIture serum decreased by 52.83%(P﹤0.01)and 53.07%(P﹤0.01), respectiveIy,when HSA-IFNα-2b 500 kU·L-1 was added. The activity of enhancer Ⅰ decreased by 40.04%(P﹤0.01)when HSA-IFNα-2b 500 kU·L-1 was added. CONCLUSlON The ceII modeI of HBV repIication for evaIuating anti-HBV agents is successfuIIy estabIished. HSA-IFNα-2b exhibits noticeabIe anti-HBV effect invitro.

Objective To investigate the optimal concentration of manganese-enhanced MRI (MEMRI)in the visual pathway of experimental rats.Methods Sprague-Dawley rats were intravitreally injected with 3 μL of 10 - 100 mmol/L MnCl2 ,respectively. The contrast-to-noise ratio (CNR)of MEMRI for optic nerve(ON)and midbrain superior colliculus (SC)enhancement were measured at differ-ent concentrations of MnCl2 .Results The ON and SC were all clearly detected by MEMRI 24 h after unilateral intravitreal injected 10-100 mmol/L MnCl2 ,respectively.The CNR increased with the increasing concentration of MnCl2 from 10 to 50 mmol/L;But the CNR decreased from 50 to 100 mmol/L.The enhancement of superior colliculus were higher than optic nerve.Conclusion The optimal concentration of MnCl2 is 30 mmol/L(3 μL)through intravitreal injection for the rat visual pathway on 1.5T MEMRI.

Toll-like receptor 4 (TLR4), a type Ⅰ transmembrane protein, has been extensively studied in the Toll-like receptor family at present. TLR4 ligands include lipopolysaccharides ( LPS) present in the outer membrane of gram-negative bacteria and monophosphoryl lipid A ( MPLA) which is a derivative of LPS. TLR4 agonists, alone, as a major component of compound adjuvants or in combination with other TLRs agonists, have been widely used as adjuvants in various vaccines and demonstrated great potential in vaccine development. This review addressed the discovery, application, features and prospect of novel vac-cine adjuvants based on TLR4 agonists, aiming to provide reference for rational use of adjuvants and further development.

Objective To evaluate the effect of targeted monitoring and comprehensive intervention measures on reducing the occurrence of catheter-associated urinary tract infection(CAUTI)in patients in non-intensive care unit(Non-ICU).Methods In quarter 4 of 2015,patients with indwelling urinary catheter in clinical departments were conducted a baseline survey(before intervention),risk factors for CAUTI in patients were analyzed,targeted monitoring programmes and comprehensive intervention measures were initiated in 2016(after intervention),incidence of CAUTI before and after intervention was compared.Results After taking intervention measures,hand hygiene compliance rate increased from 78.51%in quarter 4 of 2015 to 92.99%in quarter 3 of 2016 and 90.73%in quarter 4 of 2016(x2=7.342,3.998,respectively,both P<0.05),the correct disposal rate of patients' urinary catheterization system increased from 72.83%in quarter 4 of 2015 to 95.44%in quarter 4 of 2016(x2=30.267,P<0.05).A total of 12 067 patients with indwelling urinary catheter were monitored,incidence of CAUTI dropped from 1.03%(24/23 313)in quarter 4 of 2015(before intervention)to 0.53%(14/26 595)in quarter 4 of 2016(after intervention),difference was statistically significant(x2=4.126,P=0.042).Conclusion Improving the quality of urinary catheterization system in patients with indwelling catheter through targeted monitoring can effectively reduce the incidence of CAUTI in patients in Non-ICU.