Objective To provide a reference for prevention of trace elements deficiency in children by investigating the distribution and variation law of Ca,Cu,Zn,Fe,Mg of children in Beijing area.Methods Five kinds of trace elements( Ca,Cu,Zn,Fe,Mg) in peripheral blood of 7972 children of Beijing Children's Hospital for health examination from 2010 to 2011 were detected by atomic absorption spectrometer.The children were Beijing urban residents.They were divided into five age groups:infancy group ( 1 - 12 months,1506 cases),toddler age group ( 1 - 2 years,2766 cases),preschool age group ( 3 - 6 years,1918 cases),school age group (7 - 11 years,1576 cases) and adolescence group ( 12 - 18 years,206 cases).According to gender,each group was sub-divided into male and female group.The distributions of the 5 kinds of trace elements were analyzed among different age and gender groups.Results With the age increasing,the trend of the levels of Ca and Cu were downward,but the levels of Zn and Fe were upward,and the level of Mg was stable.For boys and girls,the levels of Cu in infancy group were (21.90 ±2.89) and(21.25 ±2.80) μmoL/L; in toddler age group,they were (21.76 ±2.78)and (21.29 ±2.69) μmol/L; in preschool age group,they were ( 21.32 ± 2.83 ) and ( 20.88 ± 2.84 ) μ mol/L; and in school age group,they were (20.81 ± 3.02 ) and ( 20.36 ± 3.37 ) μmol/L,respectively.The distribution of Cu showed significantly different among different gender groups ( t =4.640,4.475,3.290,2.894; P ＜ 0.01 ).In adolescence group,the levels of Cu were ( 19.53 ± 2.91 ) and (20.30 ± 2.90) μmol/L for boys and girls respectively and there was no significant difference ( t =- 1.796,P ＞ 0.05 ).In each gender group,the content of Ca,Zn,Fe,and Mg was not significantly different between boys and girls.It was common for the deficiency of Zn and Fe in all ages.Their total deficiency rates were 58.9％ and 19.2％.The deficiency rates of Zn and Fe in infancy were 67.7％ and 42.3％.Conclusions The distributions of Ca,Cu,and Mg in peripheral blood of children in Beijing become reasonable.Fe and Zn are deficient in different degree,and should be rationally used according to clinical guide.

Objective To investigate the character and location of a novel gene R049 and its expressed protein in uropathogenic Escherichia coli(UPEC) strain 132 isolated in China. MethodsThe chromosome library of UPEC132 was constructed by a shotgun strategy and the sequence analysis was carried out by a high-throughput pyrophosphate sequencing. Sequence reads were assembled with the Newbler program.The characters of R049-associated specific fragment were analyzed using the bioinformatics methods. Outer and inner membrane proteins of UPEC132 were extracted and then detected by SDS-PAGE and Western blot analysis together with the whole-cell lysates. ResultsThe 169 022 bp contig containing gene R049 was obtained and its sequence was very similar to the chromosome associated sequence of UPEC strain 536. It showed that a 20 773 bp fragment including R049 replaced the pathogenicity island PAI Ⅲ536 of UPEC536 in above 169 022 bp contig. The fragment had a lower GC content (46.97％) and 16 bp direct repeats in two ends. Significantly it also was adjacented to thrW tRNA, insertion element and genes coding integrase. Thus the 20 773 bp fragment was named R049 genome island(R049-GI). There were 25 ORFs in R049-GI, and gene R049 was located in the thirteenth ORF. The results of SDS-PAGE and Western blot revealed gene R049 encoded an outer membrane protein in the size of 47.0× 103. ConclusionGene R049, encoding an outer membrane protein, was a component part of the genome island in UPEC 132 chromosome acquired by horizontal gene transfer.

Objective To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) pathway in the protective effect of remifentanil or ischemic preconditioning (IPC) against hepatic ischemia-reperfusion (I/R) injury in rats.Methods One hundred and forty-four male SD rats,weighing 200-250 g,were randomly assigned into 6 group ( n =24 each):sham operation group (group S),I/R group,remifentanil group (group R),IPC group,SB203580 (a specific p38MAPK inhibitor) + remifentanil group (group SB + R),and SB + IPC group.The animals were anesthetized with intraperitoneal 20％ urethane 1 mg/kg.Partial liver ischemia was produced by clamping the hepatic pedicle supplying left lobe and middle lobe for 30 min,followed by 120 min reperfusion.In group R,remifentanil was infused intravenously at 2μg· kg- 1 · min- 1 starting from 30 min before ischemia until the end of reperfusion.In IPC group,the rats were subjected to 3 episodes of 5 min ischemia at 5 min intervals before I/R.SB203580 0.2 mg/kg was injected intravenously at 5 min before remifentanil infusion or IPC in groups SB + R and SB + IPC,and the equal volume of normal saline was given in the other groups.Six rats in each group were selected at 30,60,90 and 120 min of reperfusion and venous blood samples were taken from inferior vena cava for measurement of serum ALT and AST activities and concentrations of TNF-a and 1L-1β.The rats were then sacrificed and liver tissues were taken for microscopic examination and determination of phosphor-p38MAPK expression by Western blot.Results Compared with group S,serum AST and ALT activities and serum levels of TNF-α and IL-1β were significantly increased at each time point (P ＜ 0.05) and pathological injury was aggravated in group I/R.Compared with group I/R,serum AST and ALT activities and serum levels of TNF-a and IL-lβ were significantly decreased and phosphor-p38MAPK expression was up-regulated at 90 min of reperfusion in groups R and IPC ( P ＜ 0.05).The serum AST and ALT activities and serum levels of TNF-α and IL-1β were significantly increased,phosphor-p38MAPK expression was down-regulated at 90 min of reperfusion ( P ＜ 0.05),and pathological injury was aggravated in group SB + R compared with group R,and in group SB + IPC compared with group IPC.Conclusion Activation of p38MAPK pathway and inhibition of inflammatory response may be involved in the mechanism by which remifentanil or IPC reduces the hepatic I/R injury in rats.

Objective To estimate the application value of colony PCR in the detection of pathogenic filamentous fungi. Methods Colony PCR was established and performed to amplify the internal transcribed spacer (ITS) region of 19 species (strains) of filamentous fungus followed by sequencing analysis. At the same time, DNA extracts from 8 of the 19 species of filamentous fungus were subjected to conventional PCR. Hha I and Hinf I endonucleases were used for restriction fragment length polymorphism (RFLP) analysis of the conventional and colony PCR products. Comparison analysis was carried out between the colony and conventional PCR. Results Of the 19 strains, 16(84.2%) yielded positive results by colony PCR; sequence analysis of the PCR products of ITS region revealed a 96% - 100% similarity with the reference sequence (NCBI database)of corresponding fungi. The amplification product length and RFLP profile of these products from the 8 species of filamentous fungus, except for those from Aspergillus nidulans, were consistent between the colony and conventional PCR. Conclusions Compared with conventional PCR, colony PCR-based detection of filamentous fungi is easy to operate, time and labor-saving, with high accuracy and reliability, and can be applied to the rapid identification of filamentous fungi.

Objective To develop a PCR-RFLP method to rapidly identify filamentous fungi causing deep infection. Methods Universal fungal primers were used to amplify the internal transcribed spacer (ITS) region of Aspergillus fumigatus, Aspergillus Bavus, Aspergillus terreus, Aspergillus niger, Aspergillus versicolor, Aspergillus nidulans, Scedosporium apiospermum and Fusarium moniliforme followed by restriction fragment length polymorphism (RFLP) analysis with restrictive endonucleases Hha I, Hae III, Hinf I, Taq I and Msp I. Then, 22 clinical and 2 environmental fungal isolates were identified with the developed PCR-RFLP method. Results The RFLP analysis of PCR products with restrictive endonucleases Hha I and Hinf I allowed discrimination of 8 filamentous fungi causing invasive infection, and it took only 1 day to carry out the whole procedure from DNA extraction to PCR and restriction digestion. The identification results of 22 clinical strains and 2 environmental isolates with this PCR-RFLP method were completely consistent with those with conventional morphological method. Conclusion PCR-RFLP analysis is an efficient method for rapid identification of cultured filamentous fungi.

Objective:To investigate the effects of enteral immunonutrition on the intestinal barrier function and immune function in patients with severe pneumonia.Methods:Ninety patients with severe pneumonia were randomly divided into experimental group (n =45) and control group (n =45).All patients were received conventional therapy.In addition,patients in experimental group were given enteral immunonutrition,while patients in control group were given regular enteral nutrition.The changes of general conditions,intestinal barrier function index and immune function index were determined before treatment,on day 5 and 10 after treatment.The time of invasive mechanical ventilation,APACHE Ⅱ score and clinical effects of two groups were determined on day 10 after treatment.Results:Compared with those before treatment,in both groups,body temperature,respiration,heart rate,white blood cell count were all significantly decreased on day 5 and 10 after treatment (P ＜ 0.05).The above parameters were significantly lower in experimental group than control group on day 10 after treatment (P ＜ 0.05).The levels of serum ET,DAO were significantly decreased on day 5 and 10 after treatment in two groups compared with those before treatment (P ＜ 0.05),and these parameters were significantly lower in experimental group than control group(P ＜0.05).The number of CD3 and CD4 positive cell and the ratio of CD4 +/CD8 + were significantly increased on day 5 and 10 after treatment in two groups when compared with those before treatment (P ＜ 0.05),and these parameters were higher in experimental group than those in control group(P ＜ 0.05).The time of invasive mechanical ventilation,APACHE Ⅱ score were lower in experimental group than those in control group on day 10 after treatment (P ＜ 0.05).The rate of clinical response were higher in the experimental group than that in the control group on day 10 after treatment (P ＜ 0.05).Conclusion:Enteral immunonutrition is more effective in protecting the intestinal barrier function,improving the immune status,enhancing the immunity,reducing the time of invasive mechanical ventilation,and achieving the clinical effects of patients with severe pneumonia.

Objective To investigate the therapeutic efficacy and safety of endoscopic resection of giant gastric stromal tumors without explicit evidence of metastases.Methods A total of 12 giant gastric stromal tumors with no evidence of metastases diagnosed by endoscopic ultrasound (EUS) and computed tomography (CT) scan were managed by endoscopic resection.Operation time,blood loss and the incidence rate of perforation were recorded respectively.The diagnoses of tissue specimens were made by pathological examination and immunohistochemistry.In order to assess local recurrence and distant metastases,endoscopy and endoscopic ultrasound follow-up examinations were performed routinely at 2,6 and 12 months,and the whole abdominal CT scan was also performed at 12 months after operation.Results Endoscopic resections were successfully performed in 10 of 12 cases (83.3％),among which,6 underwent endoscopic submucosal excavation (ESE) without unexpected perforation and 4 endoscopic full-thickness resection (EFR)with intentional perforation.The rate of intentional perforation was 33.3％ (4/12),and all the perforations could be sealed by endoscopic methods.The blood losses were all more than 100 ml,which could be controlled by argon plasma coagulation,electrocoagulation or hemostatic clips.In the 10 encapsulated tumors,8 could be smoothly removed from esophagus,whose long diameter of the minimum cross section was less than 3.5 cm,however,2 tumors whose diameters were larger than 3.5 cm were taken out after segmentation.In the 10 tissue samples,9 were confirmed as low risk GIST,1 larger than 5 cm was pathologically confirmed as high risk GIST.During 1-year follow-up,no local recurrence or peritoneal metastasis was found.2 tumors,larger than 5.0 cm,could not be removed by endoscopic methods due to uncontrolled bleeding.The rate of uncontrolled bleeding was 16.7％ (2/12).The patients were transferred to surgery,and pathologically confirmed as having high risk GIST.Conclusion For low-risk giant gastric stromal tumors whose diameters were less than 5cm without evidence of metastases,endoscopic resection is considered as a safe and effective procedure.Tumors with long diameter of the minimum cross section less than 3.5 cm are more suitable for endoscopic resection,which can be smoothly taken out through cardia.However,for high-risk GIST larger than 5.0 cm,the rate of uncontrolled bleeding is high,so endoscopic resection should be adopted with discretion.

Objective To evaluate the therapeutic value of endoscopic band ligation in gastric stromal tumors.Methods Twenty-nine patients with small gastric stromal tumors(diameter<12mm)underwent endoscopic band ligation,and were followed up with routine endoscopy and EUS.Results Twentyeight patients recovered completely,with the surface healing up 4-6 weeks(average 4.8 weeks)thereafter.The neoplasm did not slough off in one patient,so he underwent a second ligation.Hemorrhage occurred in one patient three days after ligation,but was successfully managed with hemoclips.Other patients showed no complications.All the patients were followed up for 36 to 51 months,and there was one case of recurrence.Conclusion Endoscopic band ligation is an effective and safe treatment for small GISTs of less than 12 mm.

Objecfive To investigate the association between the exon 2,3,4 of MHC class Ⅰ chain-related gene-B(MICB)and ulcerative colitis(UC)in Chinese Han.Methods Using polymerase chain reaction single-stranded conformation polymorphism,allele frequency of MICB exon 2,3 and 4 in 105 patients with UC and 213 age and sex matched healthy controls were genotyped.All of the studied individuals were Chinese Han.Results Allele frequency of MICB 0106 was increased in patients with UC as compared with normal controls(19.0%vs 8.9%,P=0.000,Pc＜0.001,OR=2.402,95%CI:1.488-3.879).The frequency of MICB 0106 was increased significantly in patients with extensive colitis (24.4%vs 8.9%,P=0.000,Pc＜0.001,OR=3.294,95%CI:1.800-6.027),moderate and severe disease(24.1%vs 8.9%,P=0.000.Pc＜0.001,OR=3.294 95%CI:1.893-5.576)and in those with extra intestinal manifestations(20.5%vs 8.9%,P=0.002,Pc=0.012,OR=2.626,95%CI:1.418-4.861).Furthermore,MICB 0106 allele was higher in frequency in the male patients with UC (22.1%vs 8.0%,P=0.001,Pc=0.006,OR=3.276,95%CI:1.737-6.178)and the patients more than 40 years old(28.8%vs 8.3%,P=0.000,Pc＜0.001,OR=4.500,95%CI:2.381-8.504)as compared with healthy controls.Conclusion MICB 0106 allele is positively associated with UC,especially with extensive colitis,moderate and severe disease,presence of extra intestinal manifestations,male gender and age of more than 40 years in Chinese Han in Hubei province.

This study is to investigate effects of genistein on rat femoral bone metabolic in vitro. Rat femoral tissues was isolated and randomly divided into two groups including control group and genistein (1 x 10(-5) mol x(-1)) group. Determinations of alkaline phosphatase (ALP) activity, calcium content and osteoprotegerin (OPG), type I-collagen (Collagen-I), RANKL, Runx-2 and bone morphogenetic protein (BMP-2) mRNA expression were done by real-time PCR. The results showed that 1 x 10(-5) mol x L(-1) genistein could increase the activity of ALP and contents of Ca, regulate bone metabolism activity of OPG, RANKL, BMP-2, Collagen-I and Runx-2 mRNA expression level. Genistein can significantly modulate bone metabolism related gene expression level of rat femoral tissue in vitro, and can increase calcium content and the activity of ALP.