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 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.

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 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

BACKGROUNDGlaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project to the suprachiasmatic nucleus. This study was to investigate the effects of chronic intraocular pressure elevation on melanopsin-containing retinal ganglion cells in rats.

METHODSChronic intraocular pressure elevation was induced in one eye of adult Wistar rats by cauterization of three episcleral veins. Intraocular pressure was measured at different intervals with a rebound tonometer. Superior collicular retinal ganglion cells were retrogradely labeled from the superior colliculus with Fluorogold. Melanopsin-containing retinal ganglion cells were visualized by free-floating immunohistochemistry on whole-mount retinas. The number of labeled superior collicular and melanopsin-containing retinal ganglion cells were counted in the sample areas on flat-mounted retinas.

RESULTSCompared with contralateral control eyes, the numbers of both superior collicular and melanopsin-containing retinal ganglion cells were significantly reduced after 12 weeks of experimental intraocular pressure elevation ((2317.41 +/- 29.96)/mm(2) vs (1815.82 +/- 24.25)/mm(2); (26.20 +/- 2.10)/mm(2) vs (20.62 +/- 1.52)/mm(2), respectively). The extent of cell loss of the two types of retinal ganglion cells was similar. However, no morphologic changes were found in melanopsin-containing retinal ganglion cells.

CONCLUSIONBoth melanopsin-containing and superior collicular retinal ganglion cells were damaged by chronic ocular hypertension, indicating that glaucomatous neural degeneration involves the non-image-forming visual pathway.

Animals ; Disease Models, Animal ; Glaucoma ; pathology ; Intraocular Pressure ; Male ; Rats ; Rats, Wistar ; Retinal Ganglion Cells ; pathology ; Rod Opsins ; analysis

Electrocardiography ; Humans ; Lung Abscess ; complications ; Male ; Middle Aged ; Pulmonary Veins ; diagnostic imaging ; Radiography ; Tachycardia, Paroxysmal ; etiology ; surgery