With comparison of three different methods for the marking of amines compound, an optimal deri vatization method was selected.5-(2-Hydroxyethyl) benzoacridine (HBA) reacts with coupling agent N,N'-carbonyldiimidazole(CDI) to form an activated amide intermediate 2-(benzoacridin) ethyl-imidazole-1-carbox-ylate(BAEIC).BAEIC, which is dual-sensitive probe, reacts preferably with amino compounds at 80 ℃ in the presence of 4-dimethylaminopyridine(DMAP) catalyst in N,N-dimethylformamide(DMF) solvent to give the corresponding sensitively fluorescent derivatives with an excitation maximum at λ_(ex) of 280 nm and an emis sion maximum at λ_(em) of 510 nm.BAEIC-amine derivatives simultaneously exhibited high ionization potential with percent ionization (changing from 5.62% to 58.08% in aqueous acetonitrile and from 2.14% to 56.58% in aqueous methanol.Derivatives were not only sensitive to fluorescence but also to MS ionizable potential.The fluorescence detection limits(5/iV = 3) were 0.12-0.59 μg/L.The online APCI-MS detection limits were 1.9-14 μg/L(S/N=5).

BACKGROUND: Moxibustion can improve the symptoms of rheumatoid arthritis and reduce inflammation, but there are no uniform operation standards. The moxibustiondistance becomes one of factor influencing the therapeutic efficacy.OBJECTIVE: To compare the effects of different moxibustion distances on rheumatoid arthritis, and to explore the optimal distance.METHODS: The 8 of 40 male Sprague-Dawley rats were randomlyselected as controls. The other 32 were used to make animal modes of collagen type Ⅱ-induced ankle arthritis, and then given moxibustion at Shenshu (BL23) and Zusanli (ST36) at an interval of 1, 2 and 3 cm, respectively, once daily, 10 minutes at each point, 6 days per course for three courses, with a course interval of 1 day. Model rats with no treatment acted as model group.RESULTS AND CONCLUSION: Compared with the control group, the toe volume, arthritis index and serum levels ofinterleukin-1β and tumor necrosis factor-α except the body mass were significantly increased in the model group (P < 0.01).After moxibustion treatment, these indexes were significantly decreased in the three treatment groups (P < 0.01), especially in the 1 and 2 cm groups (P < 0.01). Narrowed articular cavity, intra-articular inflammation and pannus formation were observed in the model group, while only moderate inflammatorycell infiltration and few pannus formation were found in the 1,2, 3 cm groups. These results indicate that moxibustion improvesjoint functions and regulates immune reaction by downregulating the levels of interleukin-1β and tumor necrosisfactor-α, as well as reducing synovial hyperplasia. In addition,the optimal distance for moxibustion is 1 or 2 cm, which is amomentous proposition to improve therapeutic efficacy.

Objective: To observe the effect of different moxibustion durations on rats with rheumatoid arthritis (RA) and to evaluate the relationship between moxibustion amount and moxibustion efficacy.Methods: Eight rats were randomly selected as a normal group from the 40 male Sprague-Dawley (SD) rats, and the other 32 rats were used to establish typeⅡ collagen-induced RA models. After successful modeling, the 32 rats were randomly divided into a model group, a moxibustion for 20 min group, a moxibustion for 40 min group and a moxibustion for 60 min group, with 8 rats in each group. Rats in the normal group did not receive modeling and moxibustion intervention; rats in the model group did not receive moxibustion after modeling; rats in the moxibustion for 20 min group, the moxibustion for 40 min group and the moxibustion for 60 min group were treated with moxibustion at Shenshu (BL 23) and Zusanli (ST 36) for 20 min, 40 min and 60 min, respectively. Six days were a course of treatment, with a total of 3-course treatments and a 1-day rest between the courses of treatment. After treatment, the serum levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α, arthritis index (AI) scores, toe volumes and pathological score of synovitis were evaluated in the rats.Results: Compared with the normal group, the serum IL-1β and TNF-α levels, and the toe volumes in the model group were increased, and the differences were statistically significant (P<0.01), before the treatment. Compared with the model group, the serum IL-1β and TNF-α levels, toe volumes and arthritis index (AI) scores were significantly decreased in the moxibustion for 20 min group, the moxibustion for 40 min group and the moxibustion for 60 min group (P<0.05 orP<0.01 ). Compared with the moxibustion for 20 min group and the moxibustion for 60 min group, serum IL-1β and TNF-α levels, toe volumes and AI scores were decreased more significantly in moxibustion for 40 min group, and the differences were statistically significant (P<0.05 orP<0.01). There were no significant differences in serum IL-1β and TNF-α levels, AI scores and toe volumes between the moxibustion for 20 min group and the moxibustion for 60 min group (allP>0.05). The synovial histopathological improvement was the most obvious in the moxibustion for 40 min group, when the synovial histopathological changes were compared among the moxibustion for 20 min group, moxibustion for 40 min group and moxibustion for 60 min group.Conclusion: The therapeutic efficacy of moxibustion for 40 min in RA rats was more significant than that of moxibustion for 20 min and moxibustion for 60 min, indicating that the duration of moxibustion is the main factor affecting its therapeutic efficacy.

OBJECTIVETo investigate the expression of peripheral blood gammadelta T cells/CD4 CD25+ regulatory T cells(Treg) and cytokines interleukin 17 (IL-17) and transforming growth factor beta1 (TGF-beta1) in patients with allergic rhinitis.

METHODSFrom March 2012 to July 2012, 32 patients with allergic rhinitis (AR group) and 20 healthy control subjects (control group) were collected. The expression of peripheral blood gammadelta T cells/Treg cells were measured by flow cytometry and the levels of IL-17 and TGF-beta1 were evaluated by ELISA. SPSS 16.0 software was used to analyze the data.

RESULTSThe percentages of gammadeltaT cells in AR group were (13.30 +/- 8.62)%, which was significantly higher (t = 5.18, P < 0.01) than those in control group (5.18 +/- 1.86)%. The percentages of Treg cells in AR group were (1.75 +/- 0.56)%, which were significantly lower (t = 7.46, P < 0. 01) than those in control group (4.76 +/- 1.74)%. The IL-17 levels in AR group were (668.55 +/- 45.15) pg/ml, which were also significantly higher (t = 8.97, P < 0.01) than those in control group (573.53 +/- 17.42) pg/ml. The TGF-beta1 levels in AR group were (0.34 +/- 0.04) pg/ml, which were also significantly lower (t = 9.51, P < 0.01) than those in control group (0.49 +/- 0.06) pg/ml. There was a negative correlation between the percentages of gammadelta T cells and Treg cells (r = -0.561, P < 0.01). There was a negative correlation between the percentages of gammadelta T cells and TGF-beta1 levels (r = -0.622, P < 0.01). A positive correlation was shown between the percentages of gammadelta T cells and IL-17 levels in AR (r = 0.469, P < 0.01). A positive correlation was shown between the percentages of Treg cells and TGF-beta1 levels in AR (r = 0.738, P < 0.01). There was no correlation between IL-17 levels and the percentages of Treg cells or TGF-beta1 levels (r value was -0.111, -0.196, all P > 0.05).

CONCLUSIONThere are imbalances of gammadelta T and Treg cells in peripheral blood of patients with allergic rhinitis. gammadelta T cells may be the main cell to produce IL-17, which may play an important role in allergic rhinitis.

Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Interleukin-17 ; metabolism ; Rhinitis, Allergic ; immunology ; metabolism ; T-Lymphocytes, Regulatory ; Transforming Growth Factor beta1 ; metabolism

BACKGROUNDMacro- and microvascular diseases are the leading cause of morbidity and mortality in diabetic patients, but their mechanisms remain unclear. Recent reports provide evidence that the levels of CD55 and CD59 are decreased in diabetic microvascular diseases. However, very little is known about the levels of CD55 and CD59, the relationship between them and carotid artery intima-media thickness, and the effects of statins on CD55 and CD59 in diabetic macrovascular diseases.

METHODSThe mean fluorescence intensity (MFI) of CD55 and CD59 expression on peripheral blood leucocyte subsets (lymphocytes, monocytes and neutrophils) was studied using flow cytometry, and carotid artery intima-media thickness was measured using B-mode ultrasonography in 23 healthy subjects (controls), 19 patients with type 2 diabetes (T2DM), and 43 patients with type 2 diabetes and macrovascular diseases (T2DM-M). The patients with T2DM-M were assigned to two subgroups based on whether statins were used: group with statins (n = 23) and group without statins (n = 20).

RESULTSCompared with the controls and T2DM, the MFI of CD55 positive neutrophils was significantly lower in T2DM-M (P = 0.049 vs controls and P = 0.033 vs T2DM); similarly, the MFI of CD59 positive monocytes was also lower in T2DM-M (P = 0.038 vs controls and P = 0.043 vs T2DM). The MFI of CD59 positive neutrophils in T2DM-M was lower than in T2DM (P = 0.032). The levels of CD55 and CD59 were negatively associated with age and blood pressure (r = -0.245 - -0.352, P = 0.041 - 0.003), but not acute-phase reactants and carotid artery intima-media thickness. The levels of CD55 and CD59 increased after treatment with statins, but the results were not significantly different (P > 0.05).

CONCLUSIONSCD55 and CD59 expressions on peripheral blood leucocytes are decreased in T2DM patients with macrovascular diseases. The results suggest that the decreased levels of complement regulatory proteins might play an important role in diabetic macrovascular diseases.

Aged ; Aged, 80 and over ; CD55 Antigens ; immunology ; CD59 Antigens ; immunology ; Diabetes Mellitus, Type 2 ; immunology ; Diabetic Angiopathies ; immunology ; Female ; Flow Cytometry ; Gene Expression Regulation ; Humans ; Leukocytes ; immunology ; Male ; Middle Aged

OBJECTIVETo study the clinicopathologic features, diagnostic criteria, differential diagnosis and treatment options of ovarian steroid cell tumor, not otherwise specified (NOS).

METHODSLight microscopy and immunohistochemical study was carried out in 8 cases of ovarian steroid cell tumor, NOS. The literature was reviewed.

RESULTSThe 7 cases of benign ovarian steroid cell tumor, NOS were composed mainly of polygonal cells with granular eosinophilic cytoplasm and larger cells with vacuolated cytoplasm. They resembled the architecture of normal adrenal gland, with formation of cell nests and trabeculae. The single case of malignant ovarian steroid cell tumor had evidence of significant cellular pleomorphism, haemorrhage and coagulative tumor necrosis. The mitotic count measured about 7 per 10 high-power fields. Immunohistochemical study showed that the tumor cells expressed calretinin and alpha-inhibin. Differential diagnosis included oxyphilic granulosa cell tumor, thecoma, Sertoli cell tumor and clear cell carcinoma. The treatment options of benign ovarian steroid cell tumor, NOS was local excision or ipsilateral salpingo-oophorectomy, while the malignant counterpart should be treated with a combination of surgery and chemotherapy, including administration of GnRH agonist.

CONCLUSIONSOvarian steroid cell tumor, NOS, is the most common type of ovarian steroid cell tumors. Most of which are associated with a benign clinical outcome. Immunohistochemistry is an important adjunct for diagnosis. The treatment options of ovarian steroid cell tumor, NOS depend on its malignant potential.

Adolescent ; Adult ; Calbindin 2 ; Diagnosis, Differential ; Female ; Granulosa Cell Tumor ; pathology ; Humans ; Inhibins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; surgery ; Ovariectomy ; methods ; Ovary ; pathology ; S100 Calcium Binding Protein G ; metabolism ; Sertoli Cell Tumor ; pathology ; Sex Cord-Gonadal Stromal Tumors ; metabolism ; pathology ; surgery ; Thecoma ; pathology ; Young Adult

BACKGROUNDDiagnosis and appropriate treatment of multidrug-resistant tuberculosis (MDR-TB) remain major challenges. We sought to elucidate that persons who share a household with drug resistance tuberculosis patients are at high risk for primary drug resistance tuberculosis and how to prevent these outbreaks.

METHODSWe used 12-locus mycobacterial interspersed repetitive unit and 7-locus variable-number tandem repeat to identify household transmission of extensively drug resistant and multiple drug resistant Mycobacterium tuberculosis in three families admitted in Shanghai Pulmonary Hospital affiliated with Tongji University. Drug susceptibility tests were done by the modified proportion method in the MGIT 960 system in the same time. Clinical data were also obtained from the subjects' medical records.

RESULTSAll of the six strains were defined as Beijing genotype by the deletion-targeted multiplex PCR (DTM-PCR) identification on the genomic deletion RD105. Strains from family-1 had the same minisatellite interspersed repetitive unit (MIRU) pattern (232225172531) and the same MIRU pattern (3677235). Strains from family-2 had the same MIRU pattern (2212261553323) and the same MIRU pattern (3685134). Strains from family-3 did not have the same MIRU pattern and they differed at only one locus (223326173533, 223325173533), and did not have the same VNTR pattern with two locus differed (3667233, 3677234).

CONCLUSIONSHousehold transmission exists in the three families. A clear chain of tuberculosis transmission within family exists. Tuberculosis susceptibility should be considered when there is more than one tuberculosis patients in a family. Household tuberculosis transmission could be prevented with adequate treatment of source patients.

Adult ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Multiplex Polymerase Chain Reaction ; Mycobacterium tuberculosis ; classification ; genetics ; pathogenicity ; Radiography ; Tuberculosis, Multidrug-Resistant ; diagnostic imaging ; transmission ; Young Adult

OBJECTIVETo investigate the distribution and clonality of T cell receptor (TCR) Vγ and Vδ subfamily in peripheral blood of patients with allergic rhinitis before and after 1 year treatment with immunotherapy.

METHODSThe specific IgE and the complementary determinant region 3 (CDR3) of TCR V γ (I-III) and Vδ(1-8) subfamily genes in mononuclear cells were amplified from 10 effective cases of allergic rhinitis before and after 1 year treatment with immunotherapy, to observe the distribution and utilization of TCR Vγ and Vδ repertoire. The positive PCR products were further labeled with RT-PCR and analyzed by gene scan technique to determine the CDR3 size and evaluate the clonality of the detectable TCR Vγ and Vδ T cells. Peripheral blood of 10 healthy adults served as controls.

RESULTSAll symptoms were significantly improved after 1 year specific immunotherapy, but no changes were seen in specific IgE [(22.89 ± 9.60) kU/L before treatment, (19.62 ± 7.63) kU/L after treatment, Z = 1.051, P > 0.05]. No statistically significant differences of expression levels of the TCR Vγ I-III subfamily genes were found between patients with allergic rhinitis normal control group (t value were -0.679, -0.516, -0.808, all P > 0.05), but significantly decreased after 1 year treatment. There were statistically significant differences of expression levels of the TCR VγI-II subfamily genes before and after treatment (t value were -2.904, -2.217, all P < 0.05). 5.30 ± 0.82, 4.90 ± 0.57 and 5.20 ± 1.40 out of TCR Vδ (1-8) subfamilies were selectively expressed in T cells in patients with allergic rhinitis before and after 1 year treatment and normal control group, predominantly for TCR Vδ 1, 2, 3 and 6. The TCR Vδ 6 subfamily was found to have statistically significant differences in these groups (Fisher's Exact Test, P < 0.05). Compared with the normal control group and the allergic rhinitis group before treatment, a significant higher frequency of Vδ 6 oligoclonal was identified in T cells in patients with allergic rhinitis after 1 year treatment.

CONCLUSIONSThere was difference in the expression levels of the TCR Vγ I-III subfamily genes and distribution and clonality of TCR Vγ and Vδ subfamily T cells in peripheral blood of patients with allergic rhinitis before and after 1 year treatment. Specific immunotherapy can be effective in alleviation of the symptom in patients with allergic rhinitis during the early stage, possibly by inducing TCR γδ T cells, especially the TCR Vδ6 subfamily, and possibly no significant relativity between symptom and specific IgE.

Adolescent ; Adult ; Case-Control Studies ; Female ; Genes, T-Cell Receptor ; Humans ; Immunoglobulin E ; blood ; Immunotherapy ; Male ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; immunology ; Receptors, Antigen, T-Cell, gamma-delta ; genetics ; immunology ; Rhinitis ; genetics ; immunology ; therapy ; Young Adult

Objective The purpose of this study was to detect serum 25-hydroxyvitamin D [25-(OH) VitD] levels in children with atopic dermatitis (AD) and to explore its association with food allergy (FA).Methods Sixty cases of infant patients with AD have been collected.The morning fasting venous blood were obtained to check the 25-(OH)VitD level in serum.Moreover,IgE in 6 kinds of common food was also tested.Multiple regression analysis was used to analyze the relation between 25-(OH)VitD level in serum and FA.The potential risks and confounding factors were adjusted.Results Among the 60 AD children,67.4％ of them had FA,and the majority of them had milk allergy (86.84％),no wheat allergy was found.There was a positive correlation between 25-(OH) VitD levels and age (r=0.46,P＜ 0.01),but no statistical correlation with total IgE levels (P＞0.05).25-(OH)VitD deficiency may significantly increase the risk of suffering from FA (OR=11.20,95％CI:1.35-73.66,P=0.023).Conclusion The decrease of the 25-(OH)VitD level in infant patients with AD is associated with FA.What's more,25-(OH)VitD deficiency may be a risk factor for increasing FA.

OBJECTIVETo investigate the characteristics of microglial activation of hippocampus in experimental epileptic rats.

METHODSMorphological changes and proliferation of OX-42 positive cells were compared at different time points after status of epilepticus (SE) in lithium-pilocarpine induced epileptic rats.

RESULTSOX-42 positive cells were activated after SE, which increased to a peak at 3-7 d and in a relatively stable state at 7-14 d; then gradually decreased after 14d and returned to slightly higher level than previously at 21 d.

CONCLUSIONInflammatory injury, microglial activation and cell proliferation are closely related after seizures, microglial activation may be an important mechanism in the inflammatory injury of epilepsy.

Animals ; Cell Proliferation ; Disease Models, Animal ; Hippocampus ; cytology ; pathology ; Male ; Microglia ; pathology ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; pathology