Objective To test the level of follicular helper T cells (Tfh) and interleukin (IL)-21,CXCL13 in the peripheral blood of patients with ankylosing spondylitis (AS),and to analyze the relationship between Tfh and clinic features and explore the possible immunological pathogenesis of AS.Methods The Tfh cells were obtained from patients and normal controls and detected by flow cytometry.While the levels of IL-21,CXCL13 in patients and normal controls were measured by enzyme-linked immunosorbent assay (ELISA) tests.Data analysis were performed by Student's t-test,Rank-sum test,Spearman's correlation test.Results The expression of CD4+CXCR5 qCOS + cells (Tfh) (mean rank 33.71) and IL-21 [(299±27) ng/L],CXCL13 [(5.8±1.0) μg/L] in the peripheral blood of AS was significantly higher than normal controls [mean rank 23.54,(176±26) ng/L,(4.2±0.8) μg/L] (Z=-2.258,t=17.221,t=6.464,all P＜0.05).It was similar in AS with peripheral joint involvement compared with AS of non-peripheral joint involvement,and there was no difference between AS patients with positive HLA-B27 and those without HLA-B27.Mean -while,no correlation was found between the expression of Tfh,IL-21,CXCL13 and level of ESR,CRP,BASDAI.And there was no significant correlation between the expression of Tfh and IL-21,CXCL13 (P＞0.05).Conclusion The expression of Tfh and the levels of IL-21,CXCL13 are increased significantly,but are not closely relatedto disease activity.These results indicate that the abnormality of Tfh may play an important role in the pathogenesis of AS.

It has gradually become a consensus in the industry that the traditional Chinese medicine gypsum should be decocted first, but the understanding of decocting method is not completely unified in the works of doctors since ancient times, and there are occasional disputes about whether it is necessary to decocting first. In this study, the phase determination, physical and chemical characterization, qualitative and quantitative analysis of inorganic and organic components of the decoctions of herbal pairs and the whole prescription Maxingshigan decoction with gypsum as the center, and the pre-decoctions and co-decoctions of them were carried out to explore the scientific connotation of the pre-decoctions of gypsum. Results show that decoction phases were different between the co-decoctions and pre-decoctions of licorice-gypsum (Gancao-Shigao, GC-SG), ephedra-gypsum (Mahuang-Shigao, MH-SG) and almond-gypsum (Xingren-Shigao, XR-SG). The results of the micromorphology, particle size and zeta potential of herbal pairs and prescription (Quanfang, QF) showed that the supramolecular particles in pre-decoctions were smaller, more uniform and more stable than the co-decoctions. The results of organic components analysis showed that different cooking methods did not change the organic composition and content. ICP-OES results showed that the content of inorganic components in pre-decoctions was higher than in co-decoctions for the same boiling time of gypsum. The IR results showed that the pre-decoctions had stronger chemical functional group effect than the co-decoctions. To sum up, compared with the co-decoction, the pre-decoction of gypsum has different phase state and chemical composition interaction, and the difference of inorganic composition is an important material basis affecting the change of phase state compared with the co-decoction. It indicates that the material basis of traditional Chinese medicine decoction is indeed different whether gypsum is decocted first or not, which can provide a basis for the clinical application of decocted gypsum.

Calpain ; genetics ; Diabetes, Gestational ; genetics ; Female ; Genotype ; Haploidy ; Humans ; Polymorphism, Single Nucleotide ; Pregnancy

Objective To investigate whether allocardiac graft acceptance in the specific NF-κB impaired mice is due to regulatory T cell(Tr) and Th17 cells.Methods Mice abdominal heterotopic cardiac transplantation was performed and then divided in to control group(BALB/c→C57BL/6) and experimental group(BALB/c→IκBα/△N-Tg).Pretransplant and at day 7,30,100 posttransplant,spleens were harvested from the IκBα△ N-Tg mice,and then the Tr were detected by the fluorescence activated cell sorter.At day 5 posttransplant,the CD4 + Th17 cells from the spleens of the two groups were examined by the FACS.Additionally,at day 3 and 5 posttransplant,IL-17 expressed in the cardiac allograft was detected by Western blot.Results In the IκBα/ N-Tg mice group,the cardiac allografts were survived more than day 100,and without obviously lymphocytes infiltration.At the day 7 and 30 posttransplant,the Tr was obviously increased(21.23 ± 3.95,23.17 ± 4.11 vs 11.64 ± 1.96,P ＜ 0.05); however,the Tr decreased at the day 100 posttransplant,and had no difference with before transplant(10.79 ±2.48 vs 11.64 ± 1.96,P ＞0.05).Compared with the control group,at day 5 posttransplant,CD4+ Th17 cells in the IκBα/N-Tg mice and IL-17 expression of the cardiac allograft were both decreased.Conclusion In the early stage after transplantation,specific T cell NF-κB impaired could abrogate the balance of the Tr and Th17 cells,and induce the T cells differentiated into Tr and inhibit the Th17 cells differentiation,and then induce tolerance.

Background Phlyctenular ophthalmia is a delayed hypersensitivity reaction to some microprotein and affected mainly by adolescent in high incidence.Objective This study was to investigate the microscopy findings of phlyctenular ophthalmia and evaluate the histological changes by laser scanning confocal microscope.Methods Twenty-nine eyes suffered from phlyctenular ophthalmia and twenty normal eyes were examined using laser scanning confocal microscope.The pictures were taken by a CCD camera.All the cases had initially chest X-ray,tuberculin test,bacterial and mycobacteria culture.Results Dendritic and inflammatory cells were increased and concentrated in conjunctiva,and epithelial cells were deformed and squamatizated.The capillaries engorged and the goblet cells were injured.The corneoscleral Vogt meshing of the phlyctenular keratitis was obscured and dendritic cells were intruded into the corneas.The corneal epithelium of phlyctenular keratitis was absent and the subepithelial nerve plexus were bended and fractured,and the dendritic and inflammatory cells were intruded.Scarring of corneal stroma was seen under the laser scanning confocal microscope.Conclusions Laser scanning confocal microscopy is valuable for basic research and clinical diagnosis of phlyctenular ophthalmia.

Objective:To compare the two-dimensional electrophoresis(2-DE) maps of rat spinal cord protein extracted by two different solution systems.Methods: Adult rat spinal cord protein was precipitated with 10% trichloracetic acid in acetone and resuspended in 8 mol/L urea plus 4%CHAPS (A solution) or, 5 mol/L urea, 2 mol/L thiourea, 2%CHAPS plus 2%SB3-10 (B solution). One hundred and fifty micrograms of protein was loaded on 18 cm IPG strip holder and run isoelectric focusing electrophoresis as the first dimension, then horizontal SDS-PAGE as the second dimension. Protein spots were visualized by silver stain.Results:There were 1 059 and 1 023 protein spots in each map, of which 790 spots were matched in two maps. There were 269 and 233 spots exclusively extracted by A and B solutions, respectively. Taken together, 1292 different spots were totally obtained by A and B solutions.Conclusion: Integrating protein spots extracted by different solution systems is beneficial for achieving intact 2-DE map of tissues.

AIM To establish an HPLC method for the simultaneous content determination of six constituents in Xuanmai Ganju Granules (Scrophulariae Radix,Ophiopogonis Radix,Glycyrrhizae Radix et Rhizoma,Platycodonis Radix).METHODS The analysis of 80％ methanol extract of this drug was performed on a 35 ℃ thermostatic ZORBAX SB-C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 210,250,278 nm.RESULTS Harpagide,liquiritin apioside,liquiritin,harpagoside,cinnamic acid and glycyrrhizic acid showed good linear relationships within the ranges of 2.177-43.539 μg/mL(r =0.999 6),1.713-34.261 μg/mL (r =0.999 5),1.946-38.916 μg/mL(r =0.999 6),2.070-41.395 μg/mL(r =0.999 7),2.06-41.2 pg/mL (r =0.999 6) and 3.623-72.454 μg/mL (r =0.999 6),whose average recoveries (RS-Ds) were96.08％ (2.1％),95.55％ (2.5％),95.04％ (2.6％),94.86％ (2.7％),95.70％ (1.9％) and 95.47％ (1.9％),respectively.CONCLUSION This simple and accurate method can be used for the quality control of Xuanmai Ganju Granules.

Licorice-gypsum (gancao-shigao, GC-SG) drug pair was used as the research object, using supramolecular chemistry to explore the scientific connotation of combining herbal medicine GC with insoluble mineral medicine SG in clinical application of traditional Chinese medicine. ① The Tyndall effect, microscopic morphology and particle size of the single and co-decocted of GC and SG were observed, the paste content and conductivity were determined, and the interaction between GC and SG was detected by isothermal titration calorimetry (ITC) and infrared absorption spectroscopy (IR). ② Calcium chloride (CaCl₂), a soluble calcium salt of equal gypsum quality, was used instead of SG with GC for co-decocting to explore the effect of calcium salt content on the water decocting, and the characteristics were combined with the Tyndall effect, microscopic morphology, paste content and conductivity. ITC and IR techniques were used to detect the interaction between the two, and the interaction between them was detected by ITC and IR. The zeta potential and ultraviolet-visible spectrophotometry (UV-vis) of GC-SG and GC-CaCl₂ co-decoction were compared, and the inorganic and organic components in the co-decoction were detected by inductively coupled plasma optical emission spectrometer (ICP-OES) and high performance liquid chromatography (HPLC). The results showed: ① Compared with the liquid phase of single decoction, GC-SG co-decoction had more obvious Tyndall effect, and showed uniform spherical nanoparticles under electron microscope. Physical characterization results such as paste content and conductivity showed that co-decoction promoted the dissolution of each other's components; ITC and IR results showed that there was strong interaction between GC and SG, which preliminatively indicated that GC and SG co-decoction promoted the formation of uniform and stable supramolecular system of traditional Chinese medicine. ② When soluble calcium salt was used to substitute insoluble SG with GC for co-decocting, a stronger but astigmatic light path appeared than single decocting solution, the zeta potential was reduced, and a large number of accumulated polymers were formed. The results of paste content and conductivity showed that the dissolution of the co-decocting component was reduced than the single decocting component. ITC, UV-vis and IR results showed that there was interaction between GC with Ca²⁺ and SG. The formation of polysink indicated that a large amount of soluble calcium salt would destroy the stability of supramolecular Chinese medicine. The results of ICP-OES and HPLC showed that the glycyrrhizic acid (GA) content of the former lower than the latter, which was related to the formation of a large number of polycondensates with the increase of Ca²⁺ concentration and the decrease of the dissolution of GA and other active ingredients. This study indicates that the compatibility of GC and SG can form a uniform and stable supramolecular system of traditional Chinese medicine. Calcium salt, the main component of SG, is taken as the starting point. Excessive soluble Ca²⁺ can promote the aggregation of active ingredients such as GA, so as to reveal the scientific connotation of the compatibility of GC and SG, an insoluble mineral medicine.

OBJECTIVETo study the effect of heme oxygenase-1(HO-1) on proteins related to apoptosis in INS-1 cells with exposure to intermittent high glucose.

METHODSINS-1 cells cultured in vitro were divided into control group, persistent high glucose group (PHG), intermittent high glucose group (IHG), CoPP + intermittent high glucose group (CoPP+IHG), and ZnPP+ intermittent high glucose group (ZnPP+IHG). After 72 h of treatment with the corresponding protocols, the cells were examined for expressions of HO-1 protein by Western blotting and for expressions of Bax and Bcl-2 by immunocytochemistry.

RESULTSIn comparison with the control group, the cells in both PHG group and IHG group showed significantly increased expressions of HO-1 (P<0.01) and decreased Bcl-2/Bax ratios (P<0.05). The cells in CoPP+ IHG group exhibited a greater HO-1 protein expression but a lower Bcl-2/Bax ratio than those in IHG group (P<0.05) The ZnPP+IHG group demonstrated opposite changes in terms of HO-1, Bax and Bcl-2 expressions compared with the CoPP+IHG group.

CONCLUSIONIntermittent high glucose can lower Bcl-2/Bax ratio in INS-1 cells, and HO-1 may protect INS-1 cells against apoptosis possibly by up-regulating the Bcl-2/Bax ratio.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; Glucose ; administration & dosage ; adverse effects ; Heme Oxygenase (Decyclizing) ; metabolism ; Islets of Langerhans ; cytology ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; bcl-2-Associated X Protein ; metabolism