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 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 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 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 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 evaluate the value of contrast-enhanced ultrasound (CEUS) in diagnosing ischemic-type biliary lesion (ITBL) and analyze the relation between the enhancement patterns of bile duct wall of ITBL and its outcome.Methods 36 patients confirmed with ITBL (24 cases),anastomotic biliary stricture (3 cases),cholangitis (4 cases),biliarysludge (1 cases),and acute rejection (4 cases),who underwent CEUS examination,were enrolled in this study.The images were retrospectively analyzed in consensus by 2 readers.After reviewing the images,the readers were asked to make a diagnosis of ITBL.The diagnostic standard was hypo-or non-enhancement of hilar bile duct wall in arterial phase on CEUS.Results The diagnostic sensitivity,specificity,accuracy,positive predictive value and negative predictive value were 66.7％,83.3％,72.2％,88.9％,and 55.6％ for reader 1;62.5％,83.3％,69.4％,88.2％,and 52.6 ％ for reader 2,respectively.The interobserver agreement was good (κ =0.83).In 24 ITBL patients,the ratio of mortality or retransplantation with non enhancing hilar bile duct wall in arterial phase was much higher than that with enhancing hilar bile duct wall (non-enhancement 54.5％,hypo-enhancement 20％,hyper-or iso-enhancement 12.5％).Conclusions CEUS had diagnostic value of ITBL.Non-enhancing hilar bile duct wall in arterial phase on CEUS predicated the poor outcome.

Objective To investigate the effects of ulinastatin pretreatment on oxidative stress response and hepatocyte growth factor (HGF) after hepatectomy in rats.Methods One hundred and twelve pathogen-free male Sprague-Dawley rats,aged 3 months,weighing 230-280 g,were randomly divided into 2 groups (n=56 each): group hepatectomy (group H) and group ulinastatin pretreatment (group U).Left and median lobe resection was performed and then liver ischemia was induced by blood flow occlusion of right and caudate lobes for 30 min,followed by perfusion in both groups.Ulinastatin 50 000 U/kg was injected intravenously at 5 min before occlusion in group U.Eight rats in each group were chosen and the blood samples were taken from the inferior vena cava for measurement of serum ALT and AST activities and HGF concentration before ischemia and at 1,6,12,24and 48 h of reperfusion.Then the right lobe were removed for determination of apoptosis,SOD and myeloperoxidase (MPO) activities,MDA content,expression of proliferating cell nuclear antigen (PCNA) and liver regeneration.Apoptotic index was calculated.Another 8 rats in each group were chosen and the 7 day survival rate was recorded.Results Compared with group H,the levels of ALT,AST,MPO aud MDA and apoptotic index were significantly decreased,and the levels of HGF and SOD,PCNA expression and liver regeneration were significantly increased at different time points in group U (P ＜ 0.05).There was no significant difference in 7 day survival rate between group H and group U (P＞ 0.05).Conclusion Ulinastatin pretreatment can strengthen liver regeneration after hepatectomy in rats,the underlying mechanism may be related to inhibition of oxidative stress response and increase in HGF production.

This study is to compare the effects of kaempferide and anhydroicaritin on biomineralization of rat osteoblasts (ROB) in vitro. Calvarias were dissected aseptically from newborn SD rats, the osteoblasts were obtained by enzyme digestion and were cultured in MEM containing 10% FBS. The medium was changed every three days, and serial subculture was performed when cells covered with 90% of the dish. Kaempferide and anhydroicaritin were separately added with final concentrations of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) and 1 x 10(-7) mol x L(-1) under the conditions of osteogenic differentiation. The proliferation was measured by MTT, and the optimal concentration was detected by the ALP activity at the 9th day after osteogenic induction culture. The osteogenic indexes of kaempferide, anhydroicaritin and control group with the optimal concentration were compared. The result showed that the anhydroicaritin at concentration of 1 x 10(-5) mol x L(-1) had significantly promoted the activity of ALP, calcium content and osteocalcin content, increased the number of CFU-F(ALP) and mineralized nodules, enhanced the mRNA level of BMP-2, OSX and Runx-2, which are key genes of osteogenic differentiation, and raised the protein content of collagen-I. However, the kaempferide group had not significantly represented the ability that promoted osteogenic differentiation of ROB. The difference of osteogenic differentiation on ROB between kaempferide and anhydroicaritin was caused by the prenyl group on C-8 of icariin.

ObjectiveTo evaluate the efficacy and safety of endoscopic submucosal dissection (ESD) for precancerous lesions and early cancer at gastroesophageal junction (GEJ) by comparing endoscopic mucosal resection (EMR) with ESD.MethodsData of patients with GEJ precancerous lesions or early cancer,who received EMR ( n =51 ) or ESD ( n =28) were reviewed to compared the en bloc resection rate,R0 resection rate,operation time,complication and recurrence rate between 2 methods.ResultsEn blcc resection and R0 resection rates of ESD group (92.9％,78.6％ respectively) were significantly higher than those of EMR group (45.1％,43.1％ respectively).Local recurrence rate in ESD group (3.6％,1/28) was significantly lower than that of EMR group ( 19.6％ ).Complications including perforation,delayed hemorrhage,stricture were not significantly different between EMR and ESD groups.Mean operation time of ESD group (64.3 ±27.1 min) was significantly longer than that of EMR group (27.6 ± 14.1 min)(P ＜0.05).ConclusionESD,with a higher cure rate and en bloc rate and a lower local recurrence rate,is superior to EMR for precancerous lesions and early cancer at GEJ.

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.