Objective To construct a new intubation method with an otoscope and establish a murine model of Acinetobacter baumannii pneumonia with this method.Methods Part I:The Hallowell Intubation Pack for mice (Braintree Scientific Inc., USA)was used to construct a new intubation method with an otoscope.Part II:Twenty-four female ICR mice were randomized into 3 groups including control (group 1),immunosuppression (group 2)and infection after immunosuppression groups (group 3),with 8 mice in each group.The mice were treated with cyclophosphamide (CTX)by peritoneal injection to posterior orbital venous plexus.The total number of white blood cells,the number of neutrophils and the percentage of neutrophils were determined.Four mice were sacrificed at 0 h and 48 h after inoculation in each group.Then the lungs from each mouse were aseptically collected for quantitative culture and histopathology.Results Part I:Ten mice were successfully intubated using the new method and none of the mice was dead.Pulmonary bacterial culture at baseline (0 h)was (2.91×107-5.32×107 )CFU/g tissue,while the mean± standard deviation was (4.05 × 107 ± 0.82 × 107 )CFU/g tissue.The results showed that this new method had a perfect repeatability.Part II:Over 48 h,2 mice were dead in group 3,while no mouse was dead in other 2 groups.For group 3,the average pulmonary bacterial culture was 4.13×107 CFU/g tissue at 0 h and reached 3.62×1010 CFU/g tissue at 48 h (increased appropriate 1 000 times,P <0.01).The histopathologic changes in lung showed local granulomas and abscess in the alveolar space.Conclusions Intubation under the guidance of otoscope had the advantages of high repeatability and easy to operate.Additionally,the method provided stable and consistent bacterial inocula into lungs.The murine model of Acinetobacter baumannii pneumonia was successfully established with a new intubation method under the guidance of otoscope.

Objective: To examine the effect of functional sports training on the development of spatial awareness in children aged 6-8 years old,to provide a reference to improve children s ability of spatial sense. Methods: A class of 125 children aged 6-8 years from first, second, and third grades of an elementary school in Zhengzhou City were conveniently selected by stratified random sampling, who were divided into the experimental group ( n =62) and the control group ( n =63) by random number tables. The experimental group received functional sports intervention for 8 weeks,3 times a week,20 min each time, and the control group received traditional sports game program. Results: After the intervention,the error values of depth perception, orientation perception, and space perception in the experimertal group of 6 and 7 year old children reduced by 1.98 cm, 2.88°, and 22.00 cm (6 year old children) and 1.61 cm, 2.34°, and 17.99 cm (7 year old children) compared with the control group, respectively. Compared with those in the control group of 8 year old after the intervention, and the differences were of statistical signifiance( t =-3.07, -2.94, -3.07 ; -3.25, -3.29, -3.15, P <0.01). There was no significant difference in the error values of depth perception, orientation perception and space perception between the experimental group and the control group after intervention ( P >0.05). In the experimental group, the error values of depth perception, orientation perception and space perception reduced by 2.30 cm, 3.88°, 28.05 cm (6 year old children), 2.16 cm, 2.15°, 17.45 cm (7 year old children) and 1.16 cm, 1.81°, 9.10 cm (8 year old children) in children aged 6-8 years after intervention, significant improvement were observed compared with before intervention ( t = 8.50 , 9.04, 7.35; 7.39, 10.30, 11.05; 4.67, 4.46, 14.14, P <0.01). Compared with before the intervention, children aged 6-8 in the control group only had significant differences in space perception( t =4.13, 6.71, 8.93, P <0.01). Conclusion Functional sports games can improve depth perception, orientation perception and spatial perception for children aged 6-8 years. It can be integrated into children s daily activities to play a positive role in promoting the healthy growth of children.

Objective To investigate the effects of simulated microgravity by RCCS on proliferation and cell cytoskeleton of human HaCaT keratinocyte. Methods The rotary cell culture system (RCCS) was used to simulate the microgravity environment, and human HaCaT keratinocytes were divided randomly(random number) into the simulated microgravity group (SMG) and normal gravity group (NG). HaCaT cells in the two groups were harvested respectively after 32, 36 and 42 h culture. The HaCaT cells proliferation and cycles were detected by flow cytometry, the concentration of hb-EGF in supernatant was detected by ELISA, and the cell cytoskeleton was observed after 42 hours' culture under laser confocal microscope with FITC-labeled technique. SPSS 23.0 statistical software was used for statistical analysis, and P <0.05 was considered statistically significant. Results The flow cytometry showed that the proportions of human HaCaT keratinocytes in G1 and G2/M phases were increased while the proportion of HaCaT cells in S stage was decreased significantly after 32, 36 and 42 h RCCSculture compared with those in the normal gravity group. The HaCaT cells in G1 stage were declined along with incubation time. ELISA results showed that the hb-EGF concentration in HaCaT supernatant under simulated microgravity culture for 24 and 36 h was lower than that in the normal control group (P<0.01). The laser confocal microscope revealed that the HaCaT fluorescence intensity was decreased,and there were disordered microfilaments, structural ambiguity, pseudopodia reduction and irregularshape among FITC-labeled HaCaT cells cultured 42 h in RSSC compared with the normal gravity group.Conclusions RCCS simulated microgravity environment could inhibit the cell cycle transformation and proliferation of human HaCaT keratinocyte, affect the keratinocyte-secreting function, and induce alterations of the cell cytoskeleton.

The rotary cell culture system(RCCS)was used to simulate the microgravity environment, and FRTL-5 cells were divided into simulated microgravity group(SMG)and normal gravity group(NG). FRTL-5 cells were harvested after treatment for 6, 12, 24, and 36 h, the cell viability was measured by MTT assay, and the cells cycles were detected by flow cytometry. The ultrastructure of FRTL-5 cells was observed under laser confocal microscope with FITC-labeled technique. The MTT assay showed that the proliferation of FRTL-5 cells was significantly inhibited after RCCS treatment for 6, 12, 24, and 36h compared with NG(P<0.05), in which the most obvious effect was observed at 24h. The flow cytometry showed that the proportion of FRTL-5 cells at G1 stage in RCCS group was increased significantly after 6, 12, 24, and 36h compared with NG(P<0.05), while the proportion of FRTL-5 cells at S stage was decreased significantly(P<0.05)except that cultured with RCCS for 6 h. The proportion of FRTL-5 cells at G2/M stage was decreased in early phase(6-12 hours)of RCCS culture, with the lowest at 12h and transient increase at 24h of RCCS culture. The laser confocal microscope revealed that there were local microfilament depolymerization, tension fibers decrease, structure disorder, cellular pseudopodia reduction, and irregular shape among FITC-labeled FRTL-5 cells cultured with RCCS for 36h. (Chin J Endocrinol Metab, 2018, 34: 598-601)

Glycosylation is one of the common post-translational modifications of proteins to regulate the ability of tumor invasion, metastasis and tumor heterogeneity by interacting with glycan-binding proteins such as lectins and antibodies. Glycan microarray can be constructed by chemical synthesis, chemical-enzyme synthesis or natural glycan releasing. Glycan microarray is an essential analytical tool to discover the interaction between glycan and its binding proteins. Here we summarize the standard techniques to construct glycan microarray for the application in cancer vaccine, monoclonal antibody and diagnostic markers.
Antibodies, Monoclonal ; Glycosylation ; Lectins/metabolism* ; Microarray Analysis ; Neoplasms ; Polysaccharides

Objective To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determination of tobramycin in human serum,and examine the utility of the method in a clinical pharmacokinetic study of tobramycin inhalation.Methods Serum samples were pretreated by solid phase extraction with tobramycin-D12 as internal standard.Chromatographic separation was performed on a TitankHilic(2.1 mm × 100 mm,3 μm)column.The mobile phase consisted of0.1％formic acid-acetonitrile and 0.1％formic acid aqueous solution at a flow rate of 0.4 mL/min.Electrospray ionization source and multiple reaction monitoring(MRM)scanning were used for monitoring the quantitative ion pairs with m/z 468.3→m/z 163.3(tobramycin)and m/z 480.6→m/z 166.2(tobramycin-D12).The established method was investigated in terms of selectivity,interaction,concomitant medication,standard curve and lower limit of quantitation,precision and accuracy,recovery,matrix effect,and stability of tobramycinin.Results The linear range of tobramycin was 0.050 0-10.0 mg/L(R2=0.999 5).The intra-and inter-batch precision was satisfactory(coefficient of variation[CV]≤3.6％).The accuracy ranged from-0.4％to 6.0％.The matrix effect factor(MF)in human serum samples(including hemolysis and lipemia)ranged from 92.2％to 94.9％(CV≤2.7％).The recovery of tobramycinin was 79.5％-81.9％in serum samples,while the recovery of internal standard was 78.9％.The analyte was stable in serum samples for 72 h at room temperature and for 274 days at-20℃/-70℃.The pharmacokinetic study of tobramycin inhalation in bronchiectasis patients showed that after continuous administration of tobramycin 300 mg twice a day to 3 patients,the mean Cmax of tobramycin was(0.72±0.61)mg/L on Day 1 and(0.76±0.73)mg/L on Day 28,respectively.The corresponding Tmax was(1.83±0.61)h and(1.50±0.50)h,respectively.Conclusions The UPLC-MS/MS method established in this study is sensitive,accurate and rapid.It is successfully applied to the clinical pharmacokinetic study of tobramycin inhalation.The method may be suitable for therapeutic drug monitoring of tobramycin in clinical practice.

Polymyxin B, which is a last-line antibiotic for extensively drug-resistant Gram-negative bacterial infections, became available in China in Dec. 2017. As dose adjustments are based solely on clinical experience of risk toxicity, treatment failure, and emergence of resistance, there is an urgent clinical need to perform therapeutic drug monitoring (TDM) to optimize the use of polymyxin B. It is thus necessary to standardize operating procedures to ensure the accuracy of TDM and provide evidence for their rational use. We report a consensus on TDM guidelines for polymyxin B, as endorsed by the Infection and Chemotherapy Committee of the Shanghai Medical Association and the Therapeutic Drug Monitoring Committee of the Chinese Pharmacological Society. The consensus panel was composed of clinicians, pharmacists, and microbiologists from different provinces in China and Australia who made recommendations regarding target concentrations, sample collection, reporting, and explanation of TDM results. The guidelines provide the first-ever consensus on conducting TDM of polymyxin B, and are intended to guide optimal clinical use.
Humans ; Anti-Bacterial Agents/therapeutic use* ; China ; Drug Monitoring/methods* ; Polymyxin B ; Practice Guidelines as Topic