This study is aiming to establish an efficient metabolite extraction method for exploration of molecular mechanisms of desiccation tolerance of the resurrection plant Boea hygrometrica using a metabolomics approach. The extracts of metabolite in B. hygrometrica using methanol solution (method A) and methanol-chloroform-water solution (method B) were analyzed by gas chromatography-mass spectrometry (GC-MS). The total numbers of chromatographic peaks, extraction efficiency, retention time and the peak stability were compared. The results showed that for fresh materials, the total chromatographic peak number of method B is more than that of method A; the extraction efficiency of nine representative metabolites by method B is higher than that by method A; the comparison of 10 random chromatographic peaks revealed that the relative standard deviation (RSD) values of the retention time are less than 1% for both methods, whereas the RSD values of the extraction efficiency is different. The percentage of peaks that owned RSD values of the extraction efficiency higher than 10% is 50% for method A and 100% for method B. In addition, method B was also efficient for dry materials from B. hygrometrica. The number of chromatographic peaks, RSD value of retention time and extraction efficiency of dry materials was similar to that of fresh materials using method B, but decreased sharply using method A. Putting together, our study provided evidence that method B is an efficient extraction method for further analysis of metabolites from this resurrection species.
Chemical Fractionation ; methods ; Chloroform ; Gas Chromatography-Mass Spectrometry ; Magnoliopsida ; chemistry ; Metabolome ; Metabolomics ; Methanol ; Plant Extracts ; chemistry ; Solvents

Percutaneous endoscopic lumbar discectomy belongs to minimally invasive spine operation Its superiority includes smalleroperation wound,less bleeding,shorter hospital day,and earlier return to function,conpared with the traditional operation.At the same time,percutaneous endoscopic lumbar discectomy has complications,as the open operation.This paper reviews its common complications,diagnosis,prevention and control.

Objective To understand the epidemic characteristics of an outbreak of panresistance Klebsiella pneumoniae occurred between 2006 and 2009 in a university hospital of Shanghai, China. Methods A total of 57 panresistance K. pneumoniae isolates were collected from August 2006 to December 2009.Antibiotic susceptibility of the isolates were determined by Kirby-Bauer disc diffusion method and microbroth dilution (MBD). ESBLs-producing initial screen test and phenotypic confirmatory test and carbapenemase-producing modified Hodge test ( MHT) were performed to detect the resistance phenotype of the isolates. Be-ta-lactamases were studied by IEF, PCR and the product sequencing. While conjugation assay were conducted to understand the transferability of these genes. The genetic relationship between isolates was established by ERIC-PCR and multilocus sequence typing (MLST). Except for the antibiotics recommended by CLSI guideline in the routine test, the other antibiotics were added to find out the effective drugs to treat the infection. Results All 57 isolates were highly resistant to all examined antibiotics. All isolates produced ESBLs and carbapenemase. IEF revealed that each isolate produced four beta-lactamases. All isolates carried blaKPC-2,blaCTX-M-14,blaSHV12,blaTEM-1,qnrB and aac(6') - I b-cr. Forty-four of the 57 (77.2% ) isolates were successful to transfer their resistance genes to E. coli recipient J53 by conjugation assay. By RAPD, all 57 isolates were grouped into two genotypes that were further identified as members of MUST types 423 and 11.Sequence types 423(ST423) only occurred before May 2008 and ST11 occurred (52 isolates) after May 2008. Most of isolates of the outbreak were ST11 (91. 2% ). A part of isolates were susceptive to added antibiotics. Conclusion The outbreak of panresistance K.pneumoniae was caused by those isolates which carried multiple resistant genes. There is a different ability of dissemination between different ST types K. pneumoniae isolate. It was necessary to add the antibiotics to find out the effective drugs to treat the infection.

Objective To establish a new method, applying high resolution melt, to discriminate the VEB-3 hypotype from the clinical gram negative isolates. Methods From January to December 2003,292 consecutive and non-repetitive gram-negative bacteria producing VEB extended spectrum β-lactamase (ESBL) were collected. Extract the DNA of clinical gram negative isolates with phenol-chloroform. PCR was performed to amplify the VEB gene with the DNA being template. After that, we amplify the fragment of VEB gene containing the position 168. Then we detect the high resolution melt curve and analyze them. At last, we analyze the results of sequence and high resolution melt( HRM ). Results VEB-1 and VEB-3 gene are markedly different through HRM analysis. Conclusion It is accurately and quickly for us to identify the VEB-3 from other hypotype through the technology of HRM.

Objective To explore the incidence and significance of intra-and extraosseous edema accompanying benign tumors and tumor like diseases.Methods Imaging studies including MR,CT and plain radiograph were retrospectively reviewed in 245 cases of benign osseous tumors and tumor like diseases proved by pathology(borderline tumor,cases associated with pathological fractures,cases with lesions of skull were excluded from the study).The incidence of bone marrow and soft tissue edema were defined on T2WI with fat suppression on MRI in all the cases.x2 test was performed for the incidence rate of edema in the benign tumors and tumor like diseases.The pre-operative diagnoses were reviewed,and the influence of edema to the differential diagnosis between the benign and malignant was analyzed.Results The total incidence of bone marrow and soft tissue edema associated with benign tumors and tumor like diseases was 37.1％ (91/245),which included Langerhans cell histiocytosis 85.2％ (23/27),osteoblastoma85.0％ (17/20),osteoid osteoma93.6％ (29/31),ehondroblastoma 87.0％ (20/23).There was no statistically significant difference of edema incidence among the above-mentioned four diseases.(x2=6.35,P＞ 0.05).Both cases of endosteal capillary hemangioma had edema around.No edema was found around the lesions of other kinds of diseases.41 of 91 cases (45.1％) with edema were misdiagnosed as malignant tumor by MRI pre-operatively.Conclusion The marrow and soft tissue edema is a common sign in the benign bone tumor and tumor-like diseases,and it is frequently seen in Langerhans cell histiocytosis,osteoblastoma,osteoid osteoma,chondroblastoma.

Objective To determine the development characteristics of social anxiety of peasant-worker's children in Harbin,and explore the effect of gender,parents quarrel,parents divorced and maternal factors on social anxiety.The study provided a scientific basis for the intervention of social anxiety.Methods The children in grade 1-5 from 3 schools in Harbin were collected by random cluster sampling in 2012.Then the same questionnaire was used to track the peasant-worker's children in 2014.The questionnaire included general information,Social Anxiety Scale for Children (SASC),Egna Minnen av Bardndosna Uppforstran (EMBU),Acceptance Subscale and Psychological Control Scale.Results (1) The detection rates of social anxiety of peasant-worker's children in 2012 and 2014 were respectively 29.3％,30.2％,and the detection rates had no significant difference (x2=0.111,P=0.790).But there was significantly difference between the two years(2012:6.90±4.26;2014:5.88±4.20;t=3.419,P=0.001).Children of newly occurring social anxiety in high grade was fewer and the ability to symptom relief was better.There was no significant difference between boys and girls on newly occurring and symptom relief(newly occurring:x2=0.018,P=0.910;symptom relief:x2 =0.541,P=0.518).(2) In 2012,the differences of the detection rate of children's social anxiety on parents quarrel,parents divorced were not significant (parents quarrel:x2 =0.881,P=0.393;parents divorced:x2=0.246,P=0.658).In 2014,the detection rate of children's social anxiety on parents quarrel,parents divorced were significantly higher than children in normal families (parents quarrel:x2=11.758,P=0.001;parents divorced:x2 =9.907,P=0.004).The detection rate of children's social anxiety of divorced parents increased with age(x2 =4.398,P=0.041).(3) In 2012 and 2014,children' s social anxiety had positive correlation with mother excessive interference and over-protective,mother refusing and denying,mother punishment,and had negative correlation with mother acceptance (P＜ 0.05).In 2014,it had positive correlation with mother control and negative correlation with mother emotional warmth understanding (P＜0.05).(4) In 2012,the regression coefficient of mother refusing and denying was 0.186.In 2014,the regression coefficient of mother acceptance and control were-0.266,0.169.Conclusion Social anxiety can be spontaneous remission,and the ability was better in high grade.Parents quarrel,parents divorced and maternal factors can affect the development of peasant-worker's children' s social anxiety,which should be concerned.

An offline two-dimensional system combining a rat cardiac muscle cell membrane chromatography time-of-flight mass spectrometry (CMC-TOF/MS) with a high performance liquid chromatography time-of-flight mass spectrometry (HPLC-TOF/MS) was established for investigating the parent components and metabolites in rat urine samples after administration of the roots of Aconitum carmichaeli.On the basis of the analysis of the first dimension,retention components of the mine sample were collected into 30 fractions (one fraction per minute).Then offline analysis of the second dimension was carried out.34 compounds including 24 parent alkaloids and 10 potential metabolites were identified from the dosed rat urine,and then binding affinities of different compounds on cell membranes were compared and influences of some functional groups on activity were estimated with the semi-quantification and curve fitting method.As a result,binding affinities decreased along with the process of deacylation,debenzoylation and demethylation,which may be related to the alleviation of toxicity in the procedure of herb processing or metabolism.Moreover,some minor components in rat urine (Songorine,14-benzoylneoline,Deoxyaconitine,etc.) exerted relatively strong affinity on cell membranes are worth exploring.The results delivered by the system suggest that the CMC can be applied to in vivo study.

Objective To observe the effect of cistanche deserticola polysaccharides on learning and memory ability of different types of spatial learning and memory abilities in micey.Methods The 120 mice were randomly divided into blank group, model group, Cistanche deserticola polysaccharide, low and medium dose group, and Lacita group, 20 rats in each group. The low, medium and large dosage groups were given 25, 50 and 100 mg/kg Cistanche deserticola polysaccharides, piracetam group was given 10 mg/kg Laci Staw, blank group and model group were given 10 mg/kg volume distilled water. Continuous administration for 6 weeks. Cicloheximide mice were used to consolidate memory impairment model, using ethanol to establish a model with reappearance of memory disorders. The ability of learning and memory in water maze test in mice, the mice were detected in the brain of total protein, MDA, SOD.Results Compared with model group, the escape latency (9.45 ± 2.86 s, 12.73 ± 10.89 svs. 48.15 ± 30.33 s), and the first time arrived at the station (19.33 ± 3.27 s, 13.81 ± 9.79 svs. 40.71 ± 16.76 s) was the median, large dose group were significantly shortened (P<0.05); the first time to reach the site (11.58 ± 7.04 svs. 40.71 ± 16.76 s) in the low dose group was significantly shortened (P<0.05), the number of crossing the platform (5.46 ± 2.09vs. 3.03 ± 1.47) in the low dose group significantly increased (P<0.05); the total protein content in brain tissue (0.76 ± 0.25 g/Lvs. 0.55 ± 0.12 g/L) in the high dose group significantly increased (P<0.05). Compared with the ethanol model group, the escape latency (22.67 ± 18.35 s, 22.15 ± 16.22 s, 18.00 ± 13.44 svs. 51.33 ± 22.19 s), the first time arrived at the station (16.70 ± 11.25 s, 19.75 ± 14.62 s, 9.47 ± 5.46 svs. 30.09 ± 13.63 s) in the low, medium and high dose group were significantly shortened (P<0.05), crossing the target (5.15 ± 1.28, 4.83 ± 0.75vs. 1.34 ± 0.83) in the low, medium and high dose group were significantly increased (P<0.05).Conclusions Cistanche polysaccharide could significantly improve the reproducibility of memory impairment in mice.

Objective To investigate the status of disease stigma among patients with cervical cancer and explore the influencing factors. Method Two hundred and six patients with cervical cancer participated in the study by the social impact scale (SIS) and general information questionnaire and the relevant factors were analyzed. Results The average score of SIS was (68.04 ± 17.36). The influencing factors included the following factors age, marriage, income, profession, education and disease course (P<0.05). Conclusions The stigma in patients with cervical cancer is at a higher level and the influencing factors are diverse. It is significant to relieve their stigma and better their quality of life by strengthening psychological intervention and health instruction and establishing social support system.

ObjectiveTo investigate the influence of hepatitis B virus (HBV) infection on intrahepatic natural killer (NK) cells and innate lymphoid cell 22 (ILC22), and to provide theoretical and experimental bases for clarifying mechanisms of HBV infection in inducing innate immune response. MethodsA total of 10 male BALC/c aged 6-8 weeks were divided into experimental group and control group, with 5 mice in each group. The mice in the experimental group were treated with the hydrodynamic injection of normal saline containing 10 μg plasmids of complete HBV genome (the volume equaled to 9% of the body weight of the mouse) via the caudal vein, and those in the control group were only treated with normal saline. The mice were scarified 4 days later, and intrahepatic lymphocytes (IHLs) were isolated. Flow cytometry was used to determine the proportions of NK cells and ILC22 subset in IHLs, and the t-test was used for comparison between groups. ResultsHydrodynamic injection of the plasmids containing complete HBV genome induced high levels of HBsAg and HBeAg in mice, with an increase in the serum level of alanine aminotransferase. After HBV infection, the experimental group showed a significant increase in the proportion of intrahepatic NK cells compared with the control group (25.90%±4.92% vs 12.98%±2.13%, t=3.811, P=0.003), while there were no significant differences in the proportions of CD127+ and CD127- NK subsets in NK cells between the two groups. Moreover, after HBV infection, the experimental group showed a significant increase in the proportion of intrahepatic NKp46+ILC22 subset compared with the control group (36.05%±6.85% vs 10.22%±3.54%, t=7.372, P＜0001); however, there was no significant difference in the proportion of NKp46-ILC22 between the two groups. ConclusionHBV infection induces increased levels of intrahepatic NK cells and NKp46+ILC22 cells and thus promotes the innate immune response in the liver.