Objective To explore the difference of pathogenesis between chronic hepatitis B (CHB) and asymptomatic hepatitis B virus(HBV) carriers(ASCs) in children.Methods Subtracted libraries was constructed by suppression subtractive hybridization (SSH) from peripheral blood mononuclear cell (PBMC) of two groups, beta-2-microglobulin (?_2-MG) was screened as one of differentially expressed genes. Serum was collected from children with CHB and ASCs, the differential expression of ?_2-MG was confirmed by enzyme immunoassay.Results Subtractive libraries were constructed successfully, the differentially expression of ?_2-MG and lactroferrin were up-regulated in CHB children.And the differentially expression of ?_2-MG on the level of protein was confirmed.Conclusion The up-regulation of ?_2-MG in CHB children is involved in the pathogenesis mechanisms.

Objective To construct the eukaryotic vector that expressing hepatitis B virus (HBV) S and the single fragment of variety chain (ScFv) of monoclonal antiboy against cytotoxic Tlymphocyte-associated antigen-4 (CTLA-4) and to analyze the immunological activity of recombinant S-ScFv protein.Methods The oringially constructed pSect2/ScFv4F10 and pSect2/S were double enzyme digested by Sfi I and Hind Ⅲ,respectively.Then the HBV S gene was cloned into the pSect2/ScFv4F10 vector.The pSect2/ScFv4F10 and pSect2/S-ScFv4F10 were expressed in Chinese hamster ovary (CHO) cells,and the expressed proteins were verified through sodium dodecyl sulfatepolyacrylamide gelelectrophoresis(SDS-PAGE)andWesternblotting.Afterultrafiltration concentration and affinity chromatography,the biological affinity of the expressed ScFv4F10 and S-ScFv4F10 proteins were examined by competitive enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) technology.The comparison between groups was done by One-way ANOVA.ResultsThe eukaryotic expression vector of pSect2/S-ScFv4F10was successfully constructed,and relative molecular mass of the expressed protein of S-ScFv4 F10 was about 52 000 that analyzed by SDS-PAGE and Western blotting.With the fixed concentration of 4F10-mAb against CTLA-4,the A570 value of the mixed reaction with purified CTLA-4 antigen gradually increased with the decrease of ScFc fusion protein proportion; when the molar ratio of ScFv,S-ScFv4F10∶4F10=2∶1,the competitiveinhibitionratesagainst 4F10conjugatedantigenwere72.6％and64.5％,respectively.The affinity constants of association kinetics for CTLA-4 mAb,ScFv4F10 and S-ScFv4F10 with CTLA-4 antigen were 7.29 × 108 mol/L,9.52 × 106 mol/L and 2.04 × 106 mol/L,respectively,and the dissociation constants of KD were 1.40 × 10-9 mol/L,1.05 × 10-7 mol/L and 4.91 × 10-7 mol/L,respectively.ConclusionsThe eukaryotic expression vector of pSect2/S-ScFv4F10is successfully constructed,and the recombinant protein of S-ScFv4 F10 has a fairly high affinity with CTLA-4 antigen.

0.05).The activity of Na~+-K~+ ATPase in SMO group was significantly higher than that in HTK group at 72 h(P

OBJECTIVETo evaluate the therapeutic effect of Liujunzi decoction combined with Zuojin pills in treating the radioactive duodenitis and their mechanism, and compare with clinical routine acid suppressants combined with mucous membrane protective preparations to study the mechanism of their efficacy.

METHODAccording to the study of Williams J P and characteristics of duodenitis, and by reference to the radiation enteritis modeling standard, we took the lead in establishing the mouse radioactive duodenal injury model. The model mice were randomly divided into the control group (n = 26), traditional Chinese medicine (TCM) group (n = 16) and the western medicine (oral administration with famotidine 0.5 mL + almagate suspension 0.3 mL per mouse, once a day) group (n = 16). After the standard administrating, such objective indexes as general condition, weight, changes in health score, pathology and expression of inflammatory factors were observed to evaluate the efficacy.

RESULTThe radioactive duodenitis model of mice was successfully established with 12 Gy. Mice in the control group suffered from weight loss, anorexia, low fluid intake, loose stools, and occasionally mucous bloody stool, poor spirit, dim fur, lack of exercise and arch back. Mice in drug intervention groups were generally better than those in the pure irradiation group. The IL-6, IL-1beta, TNF-alpha mRNA expressions in spleen and mesenteric lymph node tissues in TCM and western medicine groups showed a declining trend compared with the control group. Their concentrations in peripheral blood serum also slightly changed. The TCM group revealed notable advantage in reducing inflammatory factors. The microscopic observation showed that a better mucosa repair in intervention groups than the pure irradiation group. The improved Chiu's scoring method showed a statistical significance in the difference between TCM and western medicine groups (P < 0.05).

CONCLUSIONLiujunzi decoction combined with Zuojin pills could treat acute radiation enteritis, regulate organic immunity, and inhibit acute injury, promote local tissue repair, with the potential to resist such adverse effects as radiation intestinal fibrosis. The regulation of inflammatory factor release is one of efficacy generation mechanisms.

Animals ; Cobalt Radioisotopes ; adverse effects ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Duodenitis ; blood ; drug therapy ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Mice ; Mice, Inbred BALB C ; Mucous Membrane ; drug effects ; radiation effects ; Radiation Injuries, Experimental ; blood ; drug therapy ; Tumor Necrosis Factor-alpha ; blood

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 This paper aims at establishing a inductively coupled plasma mass spectrometry ( ICP-MS) method for quantification and evaluation of iodine in human urine and serum in routine clinical laboratory .Methods This study was methodology validation research on iodine evaluation using ICP-MS.Ammonia, isopropanol and ultrapure water were mixed at certain ratio to dilute samples in the ratio of 1:10, and then the diluted samples were analyzed by ICP -MS.Re was used as the internal standard.And linearity, lower limit of detection, recovery, precision, accuracy, carryover and stability was evaluated thoroughly .Results of iodine of pregnant women who required iodine tests were retrospectively analyzed to evaluate the status of iodine .Results The method only needs 30s for analysis of one sample .It was sensitive with a lower limit detection of 0.87μg/L, the correlation coefficient was higher than 0.999 9 in ten measurements.The recovery in both serum and urine was approximately 100% (95.3% -109.9%). Based on the NIST standard reference material 3668 comparison, the bias was less than 4%( -0.9% -3.9%).The inter-coefficient variation (CV) for serum iodine and urine iodine was 1.2%-3.0%, 2. 0%-2.9%, respectively;and total CV for serum iodine and urine iodine were 3.0%-3.8%, 4.1%-4.9%, respectively.The mean carryover of this method was 0.03% and iodine was stable for at least one month at -20℃ and 4℃.The urine and serum iodine for pregnant women was (154.8 ±89.7) μg/L (mean ±SD),(75.8 ±21.4) μg/L, respectively.The correlation between urine and serum iodine was 0.21. Conclusion Establishe a rapid and simple ICP -MS method for urine and serum iodine measurement with high accurate and precise in routine clinical laboratory .

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.

Objective To explore the association between the infection of human cytomegalovirus(HCMV),herpes simplex virus typeⅠ(HSV-Ⅰ),HSV-Ⅱ,toxoplasma(TOX) and serum hepatitis B virus(HBV)these 5 pathogens and low body weight and pneumonia,and explore the clinical value of nested polymerase chain reaction (PCR) examining newborns infected with pathogens.Methods Forty-six newborn infants with low weight and 66 newborn infants with pneumonia were selected.And 1 mL pripheral blood of every newborn infant was drawn.Classic phenol-chloroform-isoamyl alcohol-protease digested,after neonatal serum extraction of DNA in peripheral blood through 2 pairs of pri-mers,the outer primer amplified larger DNA fragments and the inner primer amplified small fragments,in the amplified products.HCMV,HSV-Ⅰ,HSV-Ⅱ,TOX and HBV of the viral DNA in highly conservative district to design primer respectively and amplify its viral DNA,nested PCR was used to detect of these pathogens DNA in infants of low body weight and pneumonia,and to detect positive rate of infection.Screening for birth defects in infants in these virus infection.SPSS 10.0 software was used to analyze the relationship between infection of 5 pathogens.Results The infective rate of HCMV in 46 infants with low body quality was 91.3%,the infective rate of HSV-Ⅰwas 8.7%,the infective rate of HSV-Ⅱwas 15.2%,the infective rate of TOX was 8.7%,and the infective rate of HBV was 15.2%.Among 66 infants with pneumonia,the infective rate of HCMV was 83.3%,the infective rate of HSV-Ⅰwas 6.1%,the infective rate of HSV-Ⅱwas 16.7%,the infective rate of TOX was 6.1%,and the infective rate of HBV was 7.6%.The infective rate of HCMV was higher than that of other 4 pathogens,these infection rates were different statistically in these 5 kinds of pathogens(Pa=0).Conclusions Five kinds of pathogens both low pathosens screening is necessary newborns infants with low body weight and pneumonia,and for the early diagnosis and prevention of these pathogens.

The average life expectancy in Macau is ranking the second in the world, the consumption of fresh medicinal plants is a profound culture in Macau. The paper focus on the distribution of the antioxidant herbs, a comprehensive investigation and analysis the amount of the plant resources was carried out. The antioxidant activity of alcohol extracts was determined by using the DPPH method, and six kinds fresh herbs with high antioxidant free radical activity were screened out. Reference to adult daily dose of vitamin C, it is calculated that the daily dose amount of fresh herbs is less than 200 g. For the expected shortage of resources and the ecological status of Macau, we give some suggestions of herbal introduction in population ecology reconstruction.
Antioxidants ; pharmacology ; Ecosystem ; Free Radical Scavengers ; pharmacology ; Macau ; Plant Extracts ; pharmacology