Objective To investigate the effects of mild hypothermia ( MH ) on the expression of my- eloperoxidase(MPO) and cyclooxygenase 2 (COX 2) in rats after cerebral ischemia and reperfusion (CIR). Methods Forty-eight Wistar rats were randomly divided into four control groups (n=6 in each) and four MH groups (n=6 in each).CIR models were established by suture occlusion of the left middle cerebral artery.The rats in the MH groups,but not in the control groups,were treated with MH.Rats were killed at 4 h,8 h,12 h and 16 h after CIR.MPO expression was measured,along with the expression of COX 2 as measured by Western blot- ting and immunohistochemical methods.Results Compared with the control groups,MPO activity and the COX 2 expression in the cortex and striatum were significantly lower in all the MH groups at 4 h,8 h,12 h and 16 h after CIR.Conclusion MH treatment can protect neurons by decreasing MPO activity and COX 2 expression,allevia- ting inflammation and reducing secondary injuries after CIR.

Objective To investigate the relationship between the promoter of IL-12B gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without Helicobacter pylori(Hp) infection in children and adolescent.Methods Mucosal biopsies were obtained from 132 children and adolescent (patient group),including 100 children with chronic gastritis and 32 children with duodenal ulcer,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin (HE),and gastritis was graded according to the Sydney system.Serology,urease test and histology were taken to assess Hp status.Genomic DNA was obtained from peripheral blood or gastric biopsies of patients and 102 healthy children as normal control group.The promoter of IL-12B +1188A/G gene polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing.The genotype distributions and allele frequencies were compared between the study group and the normal control group,and the association of genotypes with clinicopathological features was studied.IL-12B mRNA level expressions in gastric mucosa were confirmed by reverse transcription PCR biopsy-based tests.Results The genotype distributions and allele frequencies of IL-12B +1188A/G gene polymorphisms were similar in gastric upper gastrointestinal diseases and healthy subjects.The IL-12B +1188A/G gene polymorphisms were not associated with Hp status.IL-12B+1188A/G gene polymorphisms did not affect IL-12B mRNA level expressions and were not associated with the degree of antrum chronic inflammation.Conclusions These data suggest that IL-12B+1188A/G gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.

Objective To investigate the value of Habib 4X in hepatic resection. Methods The clinical outcome of 21 patients with liver disease who received liver resection at the Tianjin Third Central Hospital from November 2009 to April 2010 were retrospectively evaluated. All the operations were carried out by using Habib 4X. Results All patients received hepatectomy, including right hepatectomy in three patients, left hepatectomy in one patient, multiple segmentectomy in nine patients, single segmentectomy in seven patients and partial liver resection in one patient. All tumors were reseeted completely. The mean operation time was (50±25) minutes and the mean blood loss was(129±117)ml. No patient was transferred to ICU. Three patients were complicated with bile leakage, one with lymphatic leakage and four with pleural effusion, and they were cured by non-surgical treatment. There were no patients with postoperative hemorrhage, incision infection or hepatic failure. No mortality was observed. The mean postoperative hospital stay was(19±14)days. Conclusions Radiofrequency energy was applied along the margins of the tumor to create zones of necrosis before resection with a scalpel, offering hepatobiliary surgeons an additional method for performing liver resections with minimal blood loss, low morbidity and mortality rates. As for malignant tumors, minor or major liver resection assisted by Habib 4X is safe, and it can reduce the chance of positive incisal margin.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Hepatitis B ; epidemiology ; Humans ; Infant ; Middle Aged ; Risk Factors ; Seroepidemiologic Studies ; Young Adult

Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases ; metabolism ; Bacterial Proteins ; metabolism ; Cyclization ; Cysteine Endopeptidases ; metabolism ; Enzymes, Immobilized ; metabolism ; Kinetics ; Peptides ; metabolism ; Peptides, Cyclic ; biosynthesis ; Staphylococcus aureus ; enzymology

The antibody against AT1-EC2 plays a role in some kinds of inflammatory vascular diseases including malignant hypertension, preeclampsia, and renal-allograft rejection, but the detailed mechanisms remain unclear. In order to investigate the changes of NADPH oxidase and reactive oxygen species in the aorta in a mouse model which can produce AT1-EC2 antibody by active immunization with AT1-EC2 peptide, 15 mice were divided into three groups: control group, AT1-EC2-immunized group, and AT1-EC2-immunized and valsartan-treated group. In AT1-EC2-immunized group and AT1-EC2-immunized and valsartan-treated group, the mice were immunized by 50 μg peptide subcutaneously at multiple points for 4 times: 0, 5, 10, and 15 days after the experiment. In AT1-EC2-immunized and valsartan-treated group, valsartan was given at a dose of 100 mg/kg every day for 20 days. After the experiment, the mice were sacrificed under anesthesia and the aortas were obtained and frozen in liquid nitrogen for the preparation of frozen section slides and other experiments. The titer of AT1-EC2 was assayed by using ELISA. The level of NOX1 mRNA in the aorta was determined by using RT-PCR. The expression of NOX1 was detected by using Western blotting. Confocal scanning microscopy was used to assay the α-actin and NOX1 expression in the aortic tissue. The O(2)∸ production was detected in situ after DHE staining. The mice produced high level antibody against AT1-EC2 in AT1-EC2-immunized group and AT1-EC2-immunized and valsartan-treated group, and the level of NOX1 mRNA in the aortic tissues was 1.6±0.4 times higher and the NOX1 protein expression was higher in AT1-EC2-immunized group than in control group. There were no significant differences in the level of NOX1 mRNA and protein expression between control group and AT1-EC2-immunized and valsartan-treated group. The expression and co-localization of α-actin and NOX1 in AT1-EC2-immunized group increased significantly as compared with those in control group, and the O(2)∸ production increased about 2.7 times as compared with control group. There were no significant differences between control group and AT1-EC2-immunized and valsartan-treated group. It is concluded that active immunization with AT1-EC2 can activate NOX1-ROS, and increase vascular inflammation, which can be inhibited by AT1 receptor blocker valsartan. This may partially explain the mechanism of the pathogenesis of inflammatory vascular diseases related to antibody against AT1-EC2.

Objective To investigate the association of platelet activating factor acetylhydrolase(PAF-AH) activity in children with primary nephrotic syndrome(PNS).Methods The plasma PAF-AH activity was measured in 78 children with PNS who were divided into 3 groups:steroid-responsive nephritic,steroid-dependent nephritic,steroid-resistent nephritic,after they had been given steroid for 6 months.The plasma PAF-AH activity were also measured in 60 healthy children at the same age,with spectrophotometric assay,at the ame time,the blood cholesterol was measured.Results The blood cholesterol has positive correlation with the plasma PAF-AH activity,there was no significant difference of the blood cholesterol among 3 groups in nephrotic syndrome children,there was a significant difference in the plasma PAF-AH activity among 3 groups in PNS children,but there was no significant difference in the plasma PAF-AH activity between the groups of steroid-responsive nephritic and healthy children.Conclusion Plasma PAF-AH activity is related to the sensibility to steroid treatment in children′s PNS,and the plasma PAF-AH activity in steroid-resistent nephritic is higher than steroid-dependent nephritic.It is a question that if gene mutation is related with PAF-AH activity.

Objective To release the correlation of point mutation of platelet activating factor acetylhydrolase(PAF-AH)gene and primary nephritic syndrome (PNS).Method According to the effect of hormonal therapy,94 children with PNS were divided into three groups:steroid-sensitive nephritic syndrome(SSNS),steroid-resistent nephritic syndrome(SRNS),steroid-dependent nephritic syndrome(SDNS).The point mutation of PAF-AH gene (G994T) were identified by molecular biology technique in children with PNS and 239 healthy children were set as control group.Results No statistics differences were found relating to the genotype and allele frequencies between patients with PNS,SSNS,SRNS and normal controls.But it is confirmed that the genotype and allele frequencies among patients with nephritic type nephritic syndrome (NTNS)was higher than patients with simple type nephritic syndrome(STNS) and normal controls.SDNS was higher than both SSNS and normal controls.The number of relapses during the first year after onset was significantly higher in the patients who were heterozygous for the mutant allele (GT) or homozygotes (TT) than in those of the GG homozygotes.Conclusion Most PNS children with PAF-AH gene mutation occurred at position 994 were NTNS.The risk of relapse during the treatment period was higher in patients with PAF-AH gene mutation occurred at position 994.

A test was conducted to examine the effect of several electron donors such as glucose, sodium acetate, Fe0, Fe0+glucose and Fe0+sodium acetate on reductive dechlorination of 2,4-dichlorophenol (2,4-DCP) through inoculating the unacclimated anaerobic mixed bacteria. The optimum condition and sus-tainability of Fe0 as electron donor was also been discussed. The results showed that, Fe0+glucose enhanced the dechlorination of contaminant effectively compared to glucose. Sodium acetate, Fe0 and Fe0+sodium acetate were all effective electron donors and Fe0 was the optimum, the optimum initial pH was 8.0 and quantity of added Fe0 was 2.0 g/L. 4-CP was the mainly intermediate product for 2,4-DCP dechlorination. Fe0 could support the electron for reductive dechlorination of 2,4-DCP continuously. In contrast, when so-dium acetate as electron donor, the effect of dechlorination was inferior to Fe0 with the consumption of sodium acetate.

Objective To study the relationship of urine RBC morphology between UF-100 urine sediment analytic instrument andphase contrast microscope.Methods The UF-100 urine sediment analytic instrument to analyze 500 urine specimens and study the relation-ship of urine RBC morphology between urine sediment analytic instrument and phase contrast microscope.Results The according perceptionof Normocytic,Microcytic and Non-classified RBC between phase contrast microscope and UF-100 urine sediment analytic instrument RBC-info are 91.4%,94.4%,83.3% respectively,the according perception between phase contrast microscope and RBC-P70Fsc are 94.9%,95.7%,94.7% respectively,and the according perception between phase contrast microscope and RBC Fsc-DW are 84.4%,86.8%,90.5% respectively,the specificity of UF-100 and phase contrast microscope in glomerular hematuria and non-glomerular hematuria are84.3%,88.1% and 83.3%,87.9% respectively.Conclusion The results show that the UF-100 urine sediment analytic instrument issimply operating,fast and high accurate,and which can instruct clinical dignose,therapy and prognosis judgement.