Objective To establish a simple, cost-effective method to measure reactive oxygen metabolites ( ROMs) using automatic biochemical analyzer.Methods Serum samples were collected from 50 healthy individuals and 50 hospitalized patientsin Beijing Hospital from September 2013 to November 2013.By setting up and optimizing parameters on automatic biochemical analyzer, a new method of measuring ROMS was established with TBHP as calibrator.130 healthy non-smokers were recruited in Beijing Hospital from November 2013 to December 2013 to establish reference interval, including 73 males and 57 females.The average age was 68.8 ±12.5.According to CLSI documents, there was the establishment of the new method including precision, limit of blank ( LoB ) , limit of detection ( LoD ) , limit of quantitation (LoQ), linearity,interference testing, and reference interval.Results The intra assay imprecision was 1. 5%-4.1% and the total imprecision was 4.4%-9.9%.LoB was 0.85 mmol/L TBHP;LoD was 2. 7mmol/L TBHP;LoQ was 5.9mmol/L TBHP.The linearity was 5.9-600 mmol/L (R2 =0.995).When triglyceride≤28.58 mmol/L, hemoglobin≤173 g/L, vitamin C≤60 mg/dl and bilirubin≤73.8 μmol/L, the deviation was -7.6%, 4.6%, -98.9%, 19.1%respectively.The normal reference interval was 61.9 -88.1 mmol/L TBHP.Conclusions The newly-established method has good performances of precision,LOD and linearity, which can meet the clinical needs.The interference of triglyceride and hemoglobin is small, while vitamin C and bilirubin have stronger interference on measurement.( Chin J Lab Med,2015,38:163-167)

A method was developed for determining residual narasin in chicken tissues by HPLC with post-column derivatization. The samples were extracted with iso-octane. Further cleanup was performed on LC-si cartridge after centrifugation. Then the eluent was dried by nitrogen and residues were dissolved in methanol, water mixture (90∶ 10 v/v). The samples were analyzed on an Inertsil ODS-3 C18 column with a mixture of methanol-acetic acid-water as the mobile phase and vanillin as the derivatization reagent. The detection wavelength was 520 nm. The samples were quantified with the external standard calibration curve method. The limit of detection and the limit of quantification for narasin in chicken tissues were 6.0 μg/kg and 20 μg/kg, respectively. The average recoveries of narasin in chicken tissues were 76.4 %-93.1 %, the intra-assay relative standard deviations were 2.6 %-8.9% and the inter-assay relative standard deviations were 4.7%-9.7% at spiked levels of 20-1800 μg/kg. There was a good linear correlation (the calibration coefficient is above 0.9993) between the peak areas and concentration of narasin in the range of 70-10000 μg/L.

Objective To explore the optimal diagnosis and treatment strategy of tumors at the foramen magnum,the clinical characteristics were analyzed retrospectively.Methods The clinical data of 58 cases with foramen magnum tumors treated with microsurgery in our department were collected and analyzed retrospectively.And intraoperative neurophysiological monitoring was employed in later 21 cases.Results Gross total resection was achieved in 48 of the 58 cases ; Five of the 10 patients with VA encased by tumors were totally resected.Full resection was achieved in 43 of the last 48 cases.Forty-three of the 47 cases with intradural tumors got complete resection,and total resection was achieved in 3 and 2,respectively,in 9 intra-extradural communicated tumors and 2 extradural ones.Symptoms were significantly improved in 50 cases,worsened in 5 patients,and maintained stable in the last 3 patients.Post-operative symptoms were obviously improved in the later 21 cases with intraoperative neurophysiological monitoring employed and the therapeutic effect of the treatment was much better than the former cases.Three patients died postoperatively.Fifty cases got complete follow-up from 6 months to 3 years.Among them,forty-four patients had a KPS score of 80-100,but the other 6 patients less than 80.Conclusion Microsurgical treatment for foramen magnum tumors is effective.Optimal surgical approaches and precise micro-operative techniques are crucial for the tumor removing.It is quite difficult and risky to fully resect the intra-extradural communicated tumors and the ones encasing vertebral artery.Preoperative evaluation should be profound.The application of intra-operative electrophysiological monitoring and the timely management of post-operative complications can improve the prognosis obviously.

Objective To investigate the clinical characteristics,minimal invasive operation technique and perioperative management of lateral ventrieular tumors.Methods The clinical characteristics,image diagnosis,surgical approaches and postoperative management and surgical outcomes of 65 consecutive cases of lateral ventricular tumors were retrospectively analyzed.Results In our series,total resection was achieved in 54 cases,and subtotal resection was achieved in 11 cases.Lateral ventricular tumors were mostly found in male and in the left side.Headache caused by increased intracranial pressure was the most common clinical symptom.Ependymocytoma and astrocytic glioma are the most common pathologic diagnosis.Postoperative complications included fever (26 cases),hydrocephalus (9 cases),intraventricular hematoma (7 cases) of which 2 cases were evacuated by craniotomy,epilepsy (7 cases),wound infection (3 cases).Postoperative death was happened in 3 cases.Two of them died of respiratory failure due to postoperative epilepsy.Conclusion Early discovery of lateral ventricle tumors,meticulous operation,subtle micromanipulation and right postoperative treatment are the criticality to improve the rate of total resection of lateral ventricle tumors through microsurgical treatment and reduce postoperative complications and mortality.

Objective To study morphological changes of rabbit artery endothelial cell injury and atherosclerosis caused by high fluoride and the role of selenium. Methods Twenty healthy male New Zealand white rabbits, body weight (2.0 ± 0.5)kg, were randomly divided into control group(drinking deionized water, fed basic diet), fluoride group(drinking fluoride 100 mg/L deionized water, fed basic diet), selenium group(drinking selenium 1 mg/L deionized water, fed basic diet), fluoride plus selenium group(drinking fluoride 100 mg/L deionized water, selenium 1 mg/L of deionized water, fed basic diet). The experimental period was 6 months. At 0, 3, 6 months of the experiment, serum fluorine and selenium levels were determined. At the end of the experiment,thoracic aorta was collected to observe its pathology and ultrastructural changes. Results Serum fluoride was significantly higher at the 3rd and the 6th month of experiment(all P ＜ 0.01 ) in fluoride group[ (0.589 ± 0.146),(0.772 ± 0.175)mg/L] and fluoride plus selenium group[ (0.502 ± 0.094), (0.693 ± 0.158)mg/L] than in control group[ (0.174 ± 0.002), (0.208 ± 0.031 )mg/L] and serum fluoride was significantly higher at 6 months than at 3 months(P ＜ 0.05 ) in fluoride group. Serum selenium was significantly higher at the 3rd and the 6th month of experiment (all P ＜ 0.01 ) in selenium group[ (0.252 ± 0.022), (0.319 ± 0.052)mg/L] and fluoride plus selenium group[ (0.239 ±0.016), (0.294 ± 0.018)mg/L] than in control group[(0.135 ± 0.014), (0.167 ± 0.019)mg/L], and serum selenium was significantly higher at the 6th month than at 3rd month of experiment in selenium group(P ＜ 0.05). Endothelial cell apoptosis indices were (4.92 ± 1.32)%, (30.30 ± 6.80)%, (6.57 ± 2.14)% and (14.29 ± 2.99)%, respectively in control group, fluoride group, selenium group and fluoride plus selenium group. Their main effect of fluorine and selenium was statistically significant (F = 106.833,20.082, all P ＜ 0.01 ). There were antagonistic effect between fluoride and selenium(F = 30.402, P ＜ 0.01 ). Pathological changes of rabbit aortic endothelial cells in fluoride group included endothelial with attached fibrin and red blood cells, and structural of the cells changed, with serious vascular injury; in fluoride plus selenium group apoptosis of endothelial cells decreased, with reduced number of attached red blood cells and fibrin, endothelial cell structure normal, the extent and scope of vascular damage significantly reduced. Conclusions Appropriate amount of selenium inhibits the apoptosis of endothelial cells induced by high fluoride, reduces aortic structural damage caused by high fluoride, and maintains the integrity of endothelial cells, thereby antagonizes the vascular damage and atherosclerosis induced by high fluoride.

Objective: To investigate the impact and its possible mechanism of hydrogen sulfide (H2S) on myocardial collagen remodeling in experimental rats with diabetic mellitus (DM).
Methods: Rat’s DM model was established by intraperitoneal injection of STZ at 40 mg/kg. A total of 40 SD rats were randomly divided into 4 groups:Control group, DM group, DM+NaHS group, in which NaHS worked as exogenous donor of H2S and NaHS control group. n=10 in each group, all animals were treated for 8 weeks. The cardiac collagen deposition was observed by Masson staining, protein expressions of cardiac collagen types I, III, IV and transforming growth factorβ1 (TGF-β1), connective tissue growth factor (CTGF) were examined by Western blot analysis.
Results: Compared with Control group, DM group showed increased protein expressions of cardiac collagen types I and III, up-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups. Compared with DM group, DM+NaHS group presented reduced cardiac collagen expression, decreased expression of collagen types I and III, down-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups.
Conclusion: H2S may improve the myocardial collagen remodeling in experimental DM rats, the mechanism might be related to the down-regulation of TGF-β1 and CTGF expression.

[ ABSTRACT] AIM:To explore the effects of hydrogen sulfide ( H2 S) on the myocardial fibrosis in a rat model of diabetes and its mechanism.METHODS:Single intraperitoneal injection of streptozotocin ( STZ) was utilized to establish a rat model of diabetes.Sodium hydrosulfide was used as an exogenous donor of hydrogen sulfide.Male SD rats were ran-domly divided into control group, STZ group, STZ+H2 S group and H2 S group.Eight weeks later, HE and VG staining methods were used to observe the collagen distribution and collagen volume fraction was measured by image analysis.The expression levels of type I collagen, PPARγand NF-κB in the cardiac tissues were determined by Western blotting.RE-SULTS:Compared with control group, collagen distribution and the expression levels of type I collagen and NF-κB in the cardiac tissues were markedly increased (P<0.05), while PPARγwas significantly decreased in STZ group (P<0.05), but these indexes were reversed significantly in STZ+H2S group (P<0.05).The expression levels of type I collagen, PPARγand NF-κB had no significant difference between H2 S group and control group.CONCLUSION:Hydrogen sulfide attenuates cardiac fibrosis in diabetic rats, and its mechanism may be related to PPARγ-NF-κB signaling pathway.

Objective To study the application value of 5S management for the instrument management in ENT department.Methods Totally 20 instrument nurses in ENT department implementing 5S management were enrolled into an observation group,and another 20 nurses using conventional management method were involved in a control group.The two groups were compared on instrument management quality and instrument nursing management satisfaction.Results The observation group had the qualification rate of sampled sterile items,qualification rate of instrument packaging,qualification rate of instrument cleaning,efficiency of instrument application,instrument preparedness,standardized instrument allocation,intact rate of prepared instrument as well as the familiarity with instrument management and operation significantly better than those in the control group (P＜0.05).The time consumed for adding instrument,finding instrument,environment arrangement and finding expired goods in the observation group was statistically shorter than that in the control group (P＜0.05).The times of instrument set repeated disinfection in the observation group was obviously less than those in the control group (P＜0.05),and the satisfaction over nursing management in the observation group was also significantly better than that in the control group (P＜0.05).Conclusion 5S management gains high application values for instrument management in ENT department,and thus is worthy promoting clinically.

The influenza A virus matrix protein2 gene(M2)which deleted transmembrane region was amplified by overlap extending PCR,and the multiepitope gene of hemagglutinin(HA)was PCR amplified with seven continuous synthesized segments by designing primer.The two gene segments were separately cloned into pMD18T vector to sequence analysis and prokarytic expression vector pET28a+ to construct the recombinant plasmid pET28a+M2dHA.The recombinant plasmid was transformed into E.coli BL21(DE3),and the high expression strain was obtained by screening monoclones.The recombinant protein existed as inclusion bodies,which accounted about 45% of the total cellular protein.The inclusion bodies were washed with 1% Triton X100 solution twice,and dissolved in 8 mol/L urea solution.The solution protein was purified by Ni+2 affinity chromatography,and refolded by dilution renaturation,then purified by Q Sepharose FF cation exchange column.The purity of the protein was over 90% by HPLC analysis.The result of Western blot showed it has good antigenicity and specificity.These results strongly supported for the further study of the broadspectrum influenza virus subunit vaccine.

OBJECTIVEThe SSR information in the transcriptome of Erigeron breviscapus was analyzed in this study, in order to further develop new functional genes SSR markers laid a solid foundation.

METHODSSR loci were searched in all of 52,060 unigenes by using est_timmer. Perl program and SSR primers were designed by Primer3. Furthermore, 36 pairs of primers were randomly selected for the polymorphism analysis on 13 Erigeron breviscapus plants collected from different places.

RESULTA total of 3639 SSRs were found in the transcriptome of Erigeron breviscapus, distributed in 3260 unigenes with the distribution frequency of 6.99%. Di-nucleotide repeat was the main type, account for as much as 34.41% of all SSRs, followed by mono-nucleotide (31.41%) and tri-nucleotide repeat motif (28.08%). The di-nucleotide repeat motifs of AT/AT and AC/GT were the predominant repeat types (28.71%). The tri-nucleotide repeat motifs of AAT/AT was the predominant repeat types (7.94%). For validation the availability of those SSR primers, we randomly selected 36 pairs of primers for PCR amplification. Among them, 34 pair primers (94.44%) produced clear and reproductive bands, 19 pair primers showed polymorphism (52.78%), and 13 Erigeron breviscapus plants were divided into 2 groups.

CONCLUSIONThere are numerous SSRs in Erigeron breviscapus transcriptome with high frequency and various types, this will provide abundant candidate molecular markers for genetic diversity study and genetic map in this plant.

China ; DNA Primers ; genetics ; Erigeron ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny ; Polymorphism, Genetic ; Transcriptome