The interaction of Klebsiella mytoca NG13/pMC73A with Eucalyptus were studied. The results showed that the root surface and even the inner cortex of Eucalyptus were colonized by K. oxytoca. The K. oxytoca was reisolated from rhizosphere, root surface and inner root of inoculated Eucalyptus. The inoculation with K. oxytoca stimulated the excretion of Eucalyptus root and affected the contents of amino acids, carbohydrates, phytohormones of root exudates. The seedlings of Eucalyptusto to inoculation with K. oxytoca increased growth, total dry matter and N content by 29.81% - 100.40% after 3 months in comparinson to the uninoculated seedlings.

Objective To set up an effective and low-price way for enriching dendritic cells precursor from chronic myelogeous leukemia by Percoll density gradients.Methods Peripheral blood mononuclear cells(PBMCs) were collected by separation through Ficoll-Hypaque,then PBMCs were separated by 55% Percoll gradients. The cell type and DCs expressing CD1a,CD86 were detected in the high-density group(C),low-density group(B) and non-Percoll separation group(A).Results After separation of 55% Percoll,the percentage of promyelocytes and myelocytes in group B was obviously higher than those in group A and group C(P

Long non-coding RNA (lncRNA) widely exists in living organisms.It,whose length is more than 200 bp,can regulate the genic expression in the epigenetic,transcriptional and post-transcriptional level.Therefore,it can influence cell proliferation,apoptosis,vitality,immune response and oxidative stress.In recent years,the study of lncRNA is developing rapidly in the ophthalmic research,and this article reviews the latest research progress on mechanism of lncRNA in the ophthalmology disease.

The genetic indentification of 16S rDNA or ITS, capability of phosphate-solubilization and pH of medium, and optimization of medium of some microorganism isolated from mangrove were investigated in this study. The result showed that the fungi normally had much higher capacity to dissolve the inorganic phosphate than the bacteria, the capacity of the fungi was closely correlated to the pH of medium, but the relationship was weak for the bacteria. It was illustrated by single factor experiments that the optimal carbon and nitrogen sources were maltose and urea respectively. The orthogonal design was employed in testing the optimum composition of medium composed of 5 g/L maltose, 0.05 g/L urea, 5 g/L NaCl, pH 5. In this optimal medium, the effectively enrichment of bacteria could reach up to 6.06?109 CFU/mL under 30?C for 48 hours cultivation.

Objective To identify the clinical phenotypic diagnosis and gene mutation detection of two kindreds with PS deficiency. MethodsPS: A was measured by chromogenic substrate method;TPS:Ag, FPS: Ag levels were measured by ELISA method; PS gene(PROS1 gene)was detected by amplifying 15 exons and flanking intron sequences from the propositus with PCR method. PCR products were purified and directly sequenced. Results For propositus 1,PS: A was 48.6% ,TPS: Ag was 136 mg/L, FPS : Ag was 41 mg/L, PROSI gene exon 2 was in c. Heterozygous base substitutions was detected in C121T locus, which led to Arg-1Cys (R-1C) heterozygous roissense mutation encoded in PS proteins. For propositus 2, PS: A was 29.2%, TPS: Ag was 83 mg/L, FPS: Ag was 26 mg/L, PROSI gene exon 14 was in c. Heterozygous base substitutions was identified in CI687T locus, in which Gln.522Stop heterozygous nonsense mutation was encoded in PS proteins. Conclusions c. C121T is a novel mutation locus detected in PROS1 gene. This heterozygous mutation could lead to type Ⅱ PS hereditary deficiency, while c. C1687T heterozygous mutation could bring about type Ⅰ PS hereditary deficiency.

Objective To investigate the neuroprotective effect of granulocyte colony stimulating factor (G-CSF) combined with behavior training on the learning and memory abilities of cerebral ischemic injury in rats. Methods 96 Wistar rats, 2 months old, were divided as sham group (group A), model group (group B), G-CSF group (group C) and G-CSF+training group (group D). Modified middle cerebral artery occlusion (MCAO) was used to establish a ischemia 2 h/reperfusion 24 h model to all the rats, except sham group. 6 rats were selected in each group 1, 2, 4 and 8 weeks after successfully modeled, respectively. The abilities of learning and memory were detected with the latency of shuttle test video analysis system. The pathology of the hippocampus and the expression of nerve growth factor (NGF) and G-CSF were observed with HE and immunohistochemistry staining, respectively. Results The expression of NGF and G-CSF increased in group B transiently, and increased more and longer in the group C and D, especially in group D, as the time passed (P<0.05). The latency of shuttle test increased in group B, C and D, and gently decreased as the time passed, and the decrease more to less were group D, C and B (P<0.05).Conclusion The neuroprotective effects of G-CSF combined with behavior training are strengthened and long term beneficial than G-CSF alone.

Objective:To enable the low-seniority medical personnel having the ability of disposing of difficult airway properly by mastering the knowledge of basic airway management through Airway Management Simulation Training and using all kinds of airway treatment tools.Methods:The senior medical simulation training tutors were selected, and the Airway Management Simulation Training Project Team was formed to develop the training course. Through combination of video teaching and practice of simulated teaching forms, we taught 219 trainees the airway management training course. And the feasibility and effectiveness of the course were evaluated by KE's evaluation method.Results:The course of "oriental airway simulation training" was successfully developed, and the complete course package was delivered, including bilingual airway management trainee textbook, Airway Management Simulation Training tutor manual, standardized teaching video and so on. After this simulation training, students had a good grasp of airway management skills, and more than 90.86% of the students' skills assessment resulted in more than 80 points. The overall satisfaction of the students was more than 97%, and 99% participants said that the training helped them enhance their confidence in clinical treatment, and 98% participants said that the training should be promoted among medical staff.Conclusion:The course of "oriental airway simulation training" which is made up of the combination of airway technical training, correct clinical decision-making and reality simulation, has significantly improved the airway management skills, enhanced the self-confidence of low-seniority medical staffs and improved patients' safety.

Objective:To design and construct the replication-deficient recombinant adenovirus Ad-siCTGF which can silence the expression of connective tissue growth factor (CTGF) by RNA interference and verified its function. Methods:A specific sequence, which was verified to be able to silence CTGF gene with high efficiency, was cloned into pSES-HUS vector to produce the shuttle plasmid pSES-siCTGF. The plasmid after Pme Ⅰ linearization was cotransduced with pAdEasy into BJ5183 E.coli strains to construct recombinant vector Ad-siCTGF. After linearization treatment with Pac Ⅰ enzyme digestion Ad-siCTGF was transfected into HEK293 cells via liposome mediation. The recombinant adenovirus was packaged. The titer of the Ad-siCTGF was increased after three times of cross-infection. 4T1 cells were infected with the adenovirus. The silencing efficiency was tested by real-time fluorescence quantitative (RFQ)-PCR and Western blotting.Results:Pac Ⅰ enzyme digestion electrophoresis indentified that recombinant adenovirus was successfully constructed. The titer of the recombinant adenovirus Ad-siCTGF was 2.6×10~(10) pfu/mL after amplification and purification. The CTGF mRNA and protein expression in 4T1 cells were decreased by 36.27% and 31.56%, respectively, compared with the control groups.Conclusion:The recombinant adenovirus which can silence the expression of CTGF was successfully constructed. It laid a good foundation for further investigation of the action mechanism of CTGF in tumor cells.

Objective To investigate the mechanism related to reduced antibiotic sensitivity of Acinetobacter baumannii inducted by meropenem in vitro.Methods Three strains of clinically isolated carbapenems-sensitive Acinetobacter baumannii were induced by meropenem in vitro, and the mutant strains (MS1, MS2 and MS3) were obtained.Minimal inhibitory concentrations (MICs) of antimicrobial agents to strains before and after induction were determined by automatic drug sensitivity analyzer .The homology of strains was analyzed by Enterobacterial repetitive intergenic consensus -polymerase chain reaction ( ERIC-PCR).Modified Hodge test and EDTA-Na2-double disk synergy test were used to detect carbapenemase and metallo-β-lactamase (MBL), respectively.Main carbapenemase genes were detected by PCR and followed by DNA sequencing.Expressions of adeB and outer membrane proteins in strains before and after induction were detected with fluorescence quantitative PCR and SDS -polyacrylamide gel electrophoresis , respectively.t test was used for data analysis .Results The sensitivity of mutant Acinetobacter baumannii strains to meropenem and most antibiotics was reduced , except to imipenem, amikacin and polymyxin; and the reduced sensitivity to meropenem in MS2 and MS3 was of genetic stability.ERIC-PCR showed 100%homology between the mutant strains and parental strains .Both carbapenemase and metallo -β-lactamase were negative in mutant strains and parental strains , and only OXA-51 gene was found.The expressions of adeB gene in mutant strains were 24.26 ±0.91, while those in parental strains were 22.81 ±0.38, and the difference was not significant (t =2.534, P >0.05).Outer membrane protein with molecular weight 54 000 was missing in MS1, while that with molecular weight 47 000 was missing in MS2 and MS3.Conclusion Reduced antibiotics sensitivity in meropenem -induced Acinetobacter baumannii may be correlated with the deficiency of outer membrane protein with molecular weight 47 000.

ObjectiveTo determine the changes in tumor necrosis factor-α (TNF-α) and interleukin-lβ (IL-1β)expression after bone marrow mesenchymal stem cells (BMSCs) transplantation in a rat pup model of hypoxic-ischemic brain damage (HIBD) and discuss the anti-inflammatory mechanism of BMSCs transplantation in the repair of HIBD.MethodsThree-day-old Sprague-Dawley rat pups were randomly divided into three groups: the transplantation group, in which a model of HIBD was established by ligation of left common carotid artery and hypoxia for two hours followed by the injection of 2μl BMSCs (2×105 cells) into the lateral ventricle; the HIBD model group, in which HIBD model was established and 2μl phosphate saline buffer was injected into the lateral ventricle; and the sham-operation group, in which no intervention was given. Histological changes in the brain were detected by HE staining and the number of cells positive for ectodermal dysplasia-1 (ED-1) staining, which is a specific marker for activated microglia, was detected by immunohistochemistry. The protein and mRNA levels of TNF-α and IL-1β were determined by enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction. One-way analysis of variance and LSD test were applied for statistical analysis.ResultsHE staining showed that cellular edema and necrocytosis was not observed in cerebral white matter on the 7th post-transplantation day in the transplantation group, but observed in the HIBD model group. In the sham-operation group, cerebral white matter was normal. The number of ED-1 positive cells in the transplantation group (26.3±2.5) was significantly lower than that in the HIBD model group (33.0±4.0), but higher than that in the sham-operation group (2.3±0.6) (LSD test, allP<0.05). The contents of TNF-α and IL-1β both in the transplantation group and the HIBD model group increased and peaked 24 h after transplantation, then gradually decreased, but did not reach normal levels (sham-operation group) on the 7th day. The contents of TNF-α and IL-1β in brain tissue in the HIBD model group [TNF-α: (3.03±0.10), (5.57±0.19), (7.78±0.19), (4.39±0.20), (2.70±019)μg/L; IL-1β:(293.1±7.9), (369.8±17.5), (303.6±23.9), (226.7±21.6), (183.9±33.4) ng/L] were significantly higher than those in the transplantation group [TNF-α: (2.84±0.20), (3.80±0.14), (4.63±0.17), (3.56±0.03), (1.99± 0.17)μg/L; IL-1β: (267.6±14.5), (323.5±26.9), (211.2±24.9), (140.8±7.4), (100.2±8.3) ng/L] at 6, 12, 24, 48 h and 7 days after BMSCs transplantation, respectively. The contents of TNF-α and IL-1β in the sham-operation group [TNF-α:(1.03±0.02), (1.13±0.03), (1.05±0.02), (1.09±0.02), (1.07±0.02)μg/L; IL-1β:(63.6±13.0), (64.0±11.3), (60.8±10.0), (67.9±13.5), (66.2±11.7) ng/L] were significantly lower than those in the transplantation group and HIBD model group (LSD test, allP<0.05). TNF-α and IL-1β mRNA at 24 h after transplantation in the HIBD model group (TNF-α: 2.69±0.43; IL-1β: 3.07±0.38) were significantly higher than those in the transplantation group (TNF-α: 1.61±0.29; IL-1β: 1.08±0.11) and those in the sham-operation group (TNF-α: 0.94±0.16; IL-1β: 1.08±0.11) (LSD test, allP<0.05).ConclusionsBMSCs may play an important role in the recovery of preterm HIBD and the mechanism may involve the inhibition of microglia activation and down-regulation of the expression of inflammatory factors.