Objective To construct and identify recombinant vaccine Bifwlobacterium bifidum(Bb)pGEX-Sj14-3-3 of Schistosoma japonicum(Sj). Methods Total RNA was extracted from adult Sj, antigen encoding gene Sj14-3-3 was amplified by RT-PCR and cloned into Escherichia coli (E. coli)-Bb shuttle expression vector pGEX-1λT to construct recombinant plasmid pGEX-Sj14-3-3. The recombinant plasmid was transformed into E. coli BL21 (DE3).The plasmid was extracted and identified by using BamH I and EcoR I. Then pGEX-Sjl4-3-3 was electroporated into Bb to construct recombinant Bb (pGEX-Sj14-3-3) vaccine. The extracted plasmid of the recombinant Bb (pGEX-Sj14-3-3) vaccine was identified by PCR, and the size of the products was compared with Sj14-3-3 gene of adult worms.Results Sj14-3-3 of 399 bp in length was amplified by RT-PCR. The products were digested by BamH I and EcoR I , and the fragments length of plasmid pGEX-Sj14-3-3 vector was 4947 bp, and of Sj 14-3-3 gene was 399 bp.The product of 399 bp Sj14-3-3 gene was also amplified by PCR from template of the extracted plasmid of the recombinant Bb(pGEX-Sj14-3-3 ) vaccine. The size of the product obtained was just the same as expected.Conclusion The recombinant Bb(pGEX-Sj14-3-3) vaccine of Sj is successfully constructed.

Objective To investigate the effects of recombinant vaccine Bifidobacterium bifidum (Bb) pGEX-Sj14-3-3 on splenocyte apoptosis in BALB/c mice.Methods Ninety-six BALB/c mice were randomly divided into two groups according to their body mass:per os group (PO) and intranasal immunization group (IN),with 48 mice in each group.All mice were orally and intranasally immunized with recombinant vaccine Bb(pGEX-Sj14-3-3).Four mice in each group were sacrificed on weeks 0,2,4,6,8,10,12,14,16,18,20 and 22,respectively,after immunization,and splenocytes were separated and cultured with or without ConA stimulation.The apoptotic rates of splenocytes were detected by flow cytometry.Results It showed that apoptotic level of splenocytes in both groups remarkably increased after 2-4 weeks without ConA stimulation (PO:0.069 ± 0.005,0.076 ± 0.010; IN:0.037 ± 0.002,0.075 ± 0.002),and the value reached the peak on the 4th week,and the differences were statistically significant compared with that of week 0(all P ＜ 0.05).Apoptotic level of splenocytes in both groups with ConA stimulation increased after 2-6 weeks(PO:0.089± 0.006,0.098 ± 0.010,0.060±0.007; IN:0.054 ± 0.001,0.093 ± 0.003,0.058 ± 0.012),and the value also reached the peak after 4 week,respectively.The differences were statistically significant compared with that of week 0 (all P ＜ 0.05).Apoptotic level of splenocytes in each group with ConA stimulation was significantly higher than that without ConA stimulation.Conclusion It is suspected that the recombinant vaccine Bb(pGEX-Sj14-3-3) may inhibit apoptosis of splenocytes in mice immunized orally or intranasally.

ObjectiveTo study the dynamic changes of IgG,IgG subclasses,IgE and IgA in sera of BALB/c mice immunized with recombinant Bifidobacterium bifidum (Bb) pGEX-Sj14-3-3 vaccine of Schistosoma japonicum.MethodsNinty six BALB/c mice were randomly divided into two groups:oral immunization group and intranasal immunization group,48 mice in each group.Mice were orally and intranasally immunized with recombinant Bb(pGEX-Sj14-3-3) vaccine,respectively.Four mice from each group were sacrificed,respectively,on weeks 0,2,4,6,8,10,12,14,16,18,20 and 22 after immunization and their sera from the eyeballs were collected.The levels of IgG,IgG subclasses,IgE and IgA were assayed with routine Enzyme-linked immunosorbent assay(ELISA).ResultsThe titers of IgG,IgG1,IgG2a,IgG2b,IgG3,IgE and IgA in both groups increased during the 2 - 22th,2 - 14th,2 - 22th,2 - 22th,2 - 20th,2 - 22th,2 - 22th weeks,respectively.The values reached the highest level on weeks 8,6,6,4,8,10 and 6,respectively,in the oral group,and the values were (0.065 ±0.001,0.021 ± 0.002,0.011 ± 0.001,0.015 ± 0.003,0.014 ± 0.002,0.011 ± 0.001,0.013 ± 0.002),respectively,as compared with the values on week 0(0.032 ± 0.001,0.015 ± 0.002,0.005 ± 0.002,0.005 ± 0.001,0.006 ± 0.001,0.006 ± 0.001,0.005 ± 0.001 ),the differences were statistically significant(P ＜ 0.05 or ＜ 0.01 ) except that of IgG1 and IgG2b.In the intranasal group these values reached the highest levels on weeks 4,6,4,4,8,10 and 8,respectively,and the values were (0.064± 0.003,0.022 ± 0.002,0.012 ± 0.003,0.019 ± 0.001,0.013 ± 0.001,0.015 ± 0.001,0.014 ± 0.003),respectively,as compared with the value on week 0,the differences were statistically significant(P ＜ 0.05 or ＜ 0.01 ) except that of IgG1 and IgA.ConclusionsTypes Th1 and Th2 mixed type immune responses can be induced in mice by immunization with the recombinant Bb(pGEX-Sj14-3-3) vaccine by early period of immunization (2th - 10th week).

Objective To investigate the effect of all-trans retinoic acid(atRA) on the urinary TGF?_1 excretion and glomerular lesion of diabetic rats at early stage.Methods SD rats were randomly assigned into 3 groups: atRA treated group,diabetic control group and normal control group.Streptozotocin(STZ)-induced early diabetic male SD rat were used.The atRA treated group were treated with daily subcutaneous injections atRA of 10mg/kg for 7 days(n=6),and then the excretion of urinary protein and TGF?_1 and NO level of plasma,urine and renal tissue were measured,and pathological changes of their kidneys were observed.Results The diabetic control rats showed increased urinary excretion of protein and TGF?_1 and NO level of plasma,urine and renal tissue and deposit of glomerular matrix,while atRA prevented these changes.Conclusion AtRA can prevent the development of diabetic nephropathy,which is relevant with the inhibition of secretion of TGF?_1 and NO.

Objective To evaluate the difference of carotid intimal medial thickness (IMT) among different glucose tolerance status and to investigate the association of IMT with different glucose levels of 4 time points during oral glucose tolerance test (OGTT) and the lipid metabolic indices in non-diabetic subjects. Methods Eleven normal control subjects, 69 subjects with impaired glucose regulation (IGR) newly diagnosed by OGTT (including 28 patients with non-elevated OGTT 30 min and 60 min glucose values (

Objective To study insulin sensitivity and first-phase insulin secretion in obese subjects with impaired glucose regulation (ICR) in Shanghai. Methods A total of 129 subjects [38 lean controls and 91 obese subjects with ICR including 64 isolated impaired glucose tolerance (IGT) ,8 isolated impaired fasting glucose (IFC) and 19 IFC + ICT] underwent 75 g oral glucose tolerance test and insulin-modified reduced sample number (n = 12) of Bergman's minimal model method with frequently sampled intravenous glucose tolerance test (FSIGTT). Insulin resistance was determined from the insulin sensitivity index (S1) of the FSIGTT. Insulin secretion was determined during the FSIGTT by the acute insulin response to glucose (AIRg). The disposition index (DI) , the product of AIRg and S1, was used to determine whether AIRg was adequate to compensate for insulin resistance. Results (1) Compared with normal controls, the value of S1 was significantly decreased in 3 groups with ICR (all P

Objective To observe the clinical efficacy of heat-sensitive moxibustion plus point-toward-point needling in treating poststroke strephenopodia.Method Eighty patients with poststroke strephenopodia were randomized into a treatment group intervened by heat-sensitive moxibustion plus point-toward-point needling and a control group intervened by rehabilitation, 40 cases in each group. In addition to the basic treatment, the treatment group was given heat-sensitive moxibustion plus point-toward-point needling, and the control group was given rehabilitation treatment. Holden's Functional Ambulation Classification (FAC), Fugl-Meyer Assessment (FMA) of the lower-limb motor function, and Tinetti Gait Assessment (TGA) were adopted for evaluation of the two groups, and the clinical efficacies were compared.ResultThe effective rate was 90.0% in the treatment group versus 77.5% in the control group, and the difference was statistically significant (P<0.05); after the treatment,there was a significant difference in comparing the Holden's FAC between the two groups (P<0.05); the FMA score changed significantly after the treatment in both groups (P<0.05), and there was a significant difference in comparing the FMA score between thetwo groups after the treatment (P<0.05); the TGA score changed significantly after the intervention in both groups (P<0.05), and there was a significant difference in comparing the TGA score between the two groups after the intervention (P<0.05).ConclusionHeat-sensitive moxibustion plus point-toward-point needling can produce a significant efficacy in treating poststroke strephenopodia, as it can enhance the effective rate and improve the lower-limb motor function.

OBJECTIVETo explore biological exposure markers, we investigated the chromium content in peripheral erythrocytes from occupational population with broad ranges of soluble chromate exposure, as the candidate biomarker may provide the scientific evidence for health risk assessment in occupational chromate-exposed population.

METHODSA cross-sectional study was conducted in chromate exposed workers employed at a chromate factory in a district of Jinan city, Shandong Province. The studied population contained 114 workers from different processes of the chromate plants, which included 74 males and 40 females, with an age range from 25 to 52 years old, averaging at (35.83 +/- 6.14) years old; the length of service was ranging from 1 year to 37 years, an average of (14.20 +/- 6.77) years. In addition, 30 farmers in the countryside one hundred kilometers away from the factory, without exposure to chromate matched with exposed subjects by age, gender and smoking status were identified as a control group, which included 22 men and 8 women, with age ranging from 25 years old to 47 years old, having an average age of (36.13 +/- 6.17) years old. Personal information on age, chromate exposure, medical history, smoking habit and alcohol consumption was obtained at an interview. The air concentration of personal exposure was determined by individual sampling for 8 hours per day as shift work, and chromium was assayed by atomic absorption spectrometry. The chromium content in the erythrocytes from peripheral blood was determined by graphite furnace atomic absorption spectrometry. The potential plasma reduction capacity was determined by dibenzene anthracoamid dihydrazide spectrophotometry. The content of total vitamin C and reductive ascorbic acid were determined by 2, 4-dinitrobenzene hydrazine. The data were analyzed by SPSS10.0 software for statistical significance.

RESULTS(1) The results showed that the chromium levels in erythrocytes in the exposed group [(15.79 +/- 31.01) microg/L] were significantly higher than those in the control group [(3.21 +/- 2.20) microg/L] (P < 0.01). (2) There existed a dose-response relationship between the personal airborne chromate concentration and the chromium content in erythrocytes. As airborne chromate concentration lowered to 106.00 microg/m(3), the chromium content in erythrocytes increased, depending on the air concentration of chromate. (3) Correlation analysis showed that there was a significant positive correlation between airborne chromate concentration and the chromium content in erythrocytes (P < 0.01). (4) In multiple regression analysis, it was found that the potential plasma reduction capacity and reductive ascorbic acid may be a good indicator for oxidative stress produced by chromate exposure and be used to evaluate the effects on intracellular uptake of chromium (VI).

CONCLUSIONOur findings suggested that the chromium content in erythrocytes should be used as an effective exposed biomarker in the risk assessment for occupational chromate-exposure.

Adult ; Air Pollutants, Occupational ; analysis ; Biomarkers ; blood ; Chromates ; analysis ; Chromium ; blood ; Erythrocytes ; chemistry ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; analysis

OBJECTIVETo investigate morphological changes of capillary in aging brain and explore the role of vascular factor in brain aging.

METHODSTwenty-eight brains of individuals (mean age 65 years) who died without clinical or pathological involvement of nervous system and 6 brains of Alzheimer's disease (AD) patients (mean age 83 years) were obtained at autopsy. Sections from frontal lobe, occipital lobe, striatum and hippocampus of normal subjects and sections from hippocampus of AD patients were used for hematoxylin eosin (HE), lox fast blue (LFB), toluidine blue stains and ulex europaeus agglutinin (UEA) immunostaining. After observations of morphological changes of neuron and capillary, computer-aid image analysis was performed to quantify numerical density and area density of neuron and capillary in frontal lobe, occipital lobe, putamen, CA3 sector of normal subjects and CA3 sector of AD patients. Numerical ratio and area ratio of neuron and capillary were then calculated. Correlations between neuron/capillary ratio and age were estimated using Pearson's correlation test. Difference of neuron/capillary ratio in CA3 sectors between AD patients and advanced aged normal subjects (> 75 years) was analyzed with Student's t-test.

RESULTSSeveral pathological microvascular changes, including increased tortuosity, looping, bundling, stringing, and effacement of endothelia were seen in aged subjects and more prevalent in AD patients. Numerical ratio and area ratio of neuron and capillary of frontal lobe, occipital lobe and putamen significantly increased with age in normal aging subjects.

CONCLUSIONSMorphological changes and relative decrease in number and capacity of capillary in aging brain may reduce cerebral blood flow and metabolism, and consequently result in functional impairment of aging brain. Vascular factors may play an important role in the development of brain aging.

Adult ; Aged ; Aged, 80 and over ; Aging ; Alzheimer Disease ; etiology ; pathology ; Capillaries ; anatomy & histology ; pathology ; Cell Count ; Cerebral Cortex ; blood supply ; pathology ; Female ; Frontal Lobe ; blood supply ; pathology ; Hippocampus ; blood supply ; pathology ; Humans ; Image Processing, Computer-Assisted ; Male ; Middle Aged ; Neurons ; pathology ; Occipital Lobe ; blood supply ; pathology

BACKGROUNDCholecystokinin (CCK) is one of the richest neuropeptides in the mammalian brain, which is mainly distributed in the cerebral cortex, hippocampus, thalamus and caudate-putamen. CCK is implicated in a variety of behavioral functions such as food intake, learning, memory, anxiety, pain and neuroprotection. The current research results for CCK are obtained mainly through injection of CCK peptide into the body. The key issues of whether CCK can regulate diet by a central pathway and whether there are long-term regulation effects on diet are still unresolved. In this study, the effects of CCK on food intake in transgenic mice were investigated.

METHODSTransgenic mice were created by microinjection of the PDGF-CCK construct into male pronucleus of the zygotes. The genomic phonetype of transgenic mice were identified by PCR. The expression of PDGF-CCK was analyzed by Western blotting. Body weight, plasma glucose, cholesterol and triglycerides were assayed and analyzed.

RESULTSTwo PDGF-CCK transgenic independent lines were established and exhibited a high-levels brain-specific transgene expression compared with that of nontransgenic littermate controls. The food intake of male CCK transgenic mice was decreased by 5% - 10% with the same levels of water consumed compared with wild type mice. The food intake in female mice was not obviously changed. In the transgenic mice the bodyweight was lower and plasma glucose was higher compared with the nontransgenic littermate controls.

CONCLUSIONSThe high expression of the CCK gene in the brain can decrease body weight and increase plasma glucose. The differences in food intake between the males and females require further study.

Animals ; Blood Glucose ; genetics ; physiology ; Blotting, Western ; Body Weight ; genetics ; physiology ; Brain ; metabolism ; Cholecystokinin ; genetics ; metabolism ; Cholesterol ; blood ; Eating ; genetics ; Female ; Lipase ; blood ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic