Objective To investigate the incidence of osteoporosis (OP) in patients with ankylosing spondylitis and the relationship between OP and the clinical data. Methods Serum levels of bone alkaline phosphatase (BALP) and tartrate resistant acid phosphatase 5b (TRACP5b) were detected by enzyme linked immunosorbent assay ( ELISA ) in 60 cases with ankylosing spondylitis, and it was compared with normal controls. Bone mineral density (BMD) was measured through dual-energy X-ray absorptiometry ( DXA), including lumbar ( L2 - L4), bilateral femoral neck and greater trochanter. Some clinical data was collected and analyzed at the same time. Results The incidence of OP in AS patients was 35%, and the incidence of OP in the femoral proximal end was higher than that in lumbar. Compared with normal controls[ ( 1.06 ±0. 18 )U/L ], the levels of serum TRACP5b in AS[ (1.31 ± 0. 82 )U/L] patients was significantly higher ( P <0. 05 ). The levels of serum BLAP in OP combined AS group[ ( 21.65 ± 5.41 ) U/L]were significantly lower than non-OP group[ (32. 37 ± 16. 5 ) U/L] ( P <0. 05 ). The disease duration was negatively correlated with the BMD of femoral neck ( P < 0. 01 ). Conclusions There was higher incidence of OP in AS patients, which were related with the abnormality of bone metabolism and the disease duration.Multiple factors participated in the regulation of bone metabolism of AS.

OBJECTIVE: To investigate the preventive and therapeutic effects and the possible mechanisms of Dilingdan Decoction (DLDD), a compound Chinese herbal medicine, on rats with renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction (UUO). METHODS: Sixty SD rats were randomly divided into sham-operated group, untreated group, enalapril-treated group and DLDD-treated group. Blood urea nitrogen (BUN), serum creatinine (SCr), urine protein quantization in 24 hours and pathological changes of the obstructed kidney were observed. The expressions of transforming growth factor-beta1 (TGF-beta1), alpha-smooth muscle actin (alpha-SMA), fibronectin (FN) and laminin (LN) were detected by immunohistochemical method and colored-multimedia pathological image analysis system. RESULTS: Massive inflammatory infiltrates and collagen expression in renal interstitial in the untreated group were observed on the 7th day. Compared with the sham-operated group, percentages of area of TGF-beta1, alpha-SMA, FN and LN expressions in the untreated group were markedly increased (P<0.05), while the percentage of area of interstitial fibrosis was decreased in the DLDD-treated group as compared with the untreated group (P<0.05). On the 14th day, the percentages of area of TGF-beta1, alpha-SMA, FN and LN expressions were declined in two treated groups as compared with the untreated group (P<0.05), but had no statistical difference in biochemical indicators, including BUN, SCr and 24-hour urinary protein. On the 21st day, the level of SCr and the percentage of area of TGF-beta1 expression in the DLDD-treated group were lower than those of the enalapril-treated group (P<0.05). CONCLUSION: DLDD can reduce the excretion of urinary protein and the degree of interstitial fibrosis, and significantly inhibit the expressions of TGF-beta1, alpha-SMA, FN and LN. DLDD is superior to enalapril in protecting renal function after long-time application in rats.

A thermal desorption ( TD) device was developed and coupled to gas chromatography ( GC) or gas chromatography-mass spectrometry ( GC-MS ) for the qualitative and quantitative analysis of semi-volatile organic compounds on atmospheric particulate matters ( PM ) . The TD was operated by direct heating and placed on the GC injector, leading to high heating rate and easy transfer of analytes to GC without focusing of analytes by cold trap. For establishing the TD-GC method, the materials used for supporting PM samples, temperature and time of thermal desorption, and types of sample injection were investigated for detection of sixteen polycyclic aromatic hydrocarbons ( PAHs) and nine n-alkanes. The limits of detection of the proposed TD-GC method were in the range of 0. 014-0. 093 ng for PAHs, and 0. 016-0. 026 ng for n-alkanes, respectively, with the correlation coefficients of correlation above 0. 9975. The TD-GC method was applied to the determination of trace PAHs and n-alkanes on PM10 samples from three cities. The recoveries were in the range of 95%-135% ( PAHs) and 95%-115% ( n-alkanes) , respectively. Finally, the TD was coupled to GC-MS for comparison of the contents of PAHs and n-alkanes on PMx with different particulate size ( x=10 , 5, 2, 1, 0. 5, 0. 25, 0. 1).

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.

Objective The goal of this study was to investigate the correlation between the level of serum vitamin D and bone metabolism and disease activity in ankylosing spondylitis(AS) patients,and thus to explore the role of vitamin D in bone metabolism in AS patients.Methods The serum levels of BALP,TRACP-5b,25-(OH)D3 and blood lymphocytes VDR of 80 AS patients were measured by enzyme-linked immunosorbent assay (ELISA) and compared with those of the control group.Bone mineral density (BMD)was measured by dual-energy X-ray absorptiometry (DEXA).The ESR and CRP level of AS patients were also measured.The correlation between those parameters was analyzed and evaluated.Patients were divided into normal,insufficient and deficient subgroups according to the serum 25-(OH)D3 levels for further comparison.Indepondent saimple t test,t'test andx2 test were used for statistical analysis.Results The 25-(OH)D3 of AS patients [(11.9±2.7) μg/L] was significantly lower than that of the control groups [(22.3±7.9) μg/L] (P＜0.05),while the serum levels of BALP [(3.9±2.7) μg/L] and TRACP-5b [(46±25) ng/L] of AS patients were significantly higher than those of the control group [(2.4±1.0) μg/L] (P＜0.05).According to linear correlation analysis,25-(OH)D3 was negatively correlated with CRP (r=0.324,P=0.003).The ESR,BALP,TRACP-5b in the deficient subgroup were higher than those in the normal and insufficient subgroups(P＜0.05).Conclusion The plasma 25-(OH)D3 may decrease in AS patients,and this may activate bone metabolism,results in increased morbidity of osteoporo-sis,and negatively affect disease activity.

Objective To investigate the changes and significance of Th1/Th2 cytokines in ankylosing spondyhtis (AS).Metbotis Serum and synovial fluid levels of TNF-α,IL.17.IL-10 and IL-4 were detected by en-zyme linked immunosorbent assay(ELISA)in AS cases.Results The serun levels of TNF-α,IL-17 and IL-10 were significantly higher,while the serum level of IL-4 Was significantly lower in AS group than in normal group.The levels of TNF-α.IL-17 and IL-4 in synovial fluid were significantly higher than in serum.Conclusion Disequilibrium exists in Th1/Th2 cytokine network of AS.and there is a strong predominance of Th1.

Objective To investigate the death of chondrocytes in rats which feed with T-2 toxin under selenium (Se) deficient conditions.Methods Thirty two healthy male SD rats were divided into two groups by weight which were normal diet group and Se deficiency diet group,16 rats in each group.Rats in normal diet group were fed with Se 101.5 μg/kg diet,and rats in Se deficiency diet group were fed with Se 1.1 μg/kg diet for 30 d.Normal diet group was divided into control group and T-2 toxin group,and Se deliciency diet group was randomly divided into Se-deficiency group and Se-deficiency plus T-2 toxin group,8 rats in each group.After that,rats in T-2 toxin and Se-deficiency plus T-2 toxin groups were administrated intragastrically with T-2 toxin (100 μg/kg) everyday for 30 d.Rats were put to death,the left knee was taken and stained with hematoxylin-eosin and SafraninFast green,pathological changes of rat's knee joint cartilage were observed under light microscopy,expression levels of active caspase-3 and receptor interacting protein 3 (RIP3) in rat's articular cartilage cells were determined via the immunohistochemical method.The apoptosis was also detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL).Results Red ghost outlines of chondrocyte and multiple chondral cell clusters surrounded the non-cell areas in deep zone of articular cartilage of knee joint stained with hematoxylineosin were seen in Se-deficiency plus T-2 toxin group under light microscope.In the superficial zone of cartilage,the positive percent of TUNEL and active caspase-3 in Se-deficiency plus T-2 toxin group was higher than those of control group,Se-deficiency group and T-2 toxin group [(7.47-± 0.34)％ vs (4.68 ± 0.54)％,(2.67-± 0.64)％,(2.56 ±0.54)％;(4.75 ± 0.67)％ vs (1.24 ± 0.25)％,(0.00 ± 0.00)％,(0.00 ± 0.00)％,P ＜ 0.05].In the middle zone of cartilage,the positive percent of TUNEL,active caspase-3 and RIP3 in Se-deficiency plus T-2 toxin group was significantly higher than those of control group,T-2 toxin group and Se-deficiency group [(72.06 ± 6.15)％ vs (16.10 ± 3.00)％,(19.57 ± 3.49)％,(19.33 ± 5.19)％;(51.13 ± 4.18)％ vs (10.97-± 3.01)％,(15.36 ± 4.37)％,(15.23 ± 3.13)％;(25.91 ± 13.39)％ vs (1.59 ± 1.14)％,(4.32 ± 2.91)％,(7.50 ± 5.00)％,P ＜ 0.05].The positive percents of TUNEL,active caspase-3 and RIP3 were not significantly different in the deep zone (P ＞ 0.05).Conclusion The death of the middle zone in the rat cartilage induced by T-2 toxin under selenium deficient conditions isapoptosis and necroptosis.

Objective:To study the effects of T-2 toxin on expression of fibroblast growth factor 8 (FGF8) and fibroblast growth factor receptor 3 (FGFR3) in articular cartilage and subchondral marrow of rats under low selenium condition, and to explore the mechanism of deep cartilage injury and secondary complications in Kaschin-Beck disease (KBD).Methods:Twenty-four healthy male SD rats weighted 60 - 80 g were selected, they were divided into conventional feed group (selenium content of 101.5 μg/kg) and low-selenium feed group (selenium content of 1.1 μg/kg) by random number table method, with 12 rats in each group. After 30 days of feeding, the conventional feed group was further divided into control group and T-2 toxin group (100 μg·kg -1·d -1), and the low-selenium feed group was further divided into low-selenium group and low-selenium+ T-2 toxin group, with 6 rats in each group. After 30 days of feeding, the rats were sacrificed and the knee cartilage with cancellous bone was taken. Pathological changes of knee cartilage were observed by HE staining. Immunohistochemical method was used to detect the expression of FGF8 and FGFR3 in cartilage and subchondral marrow of knee joint, positive expression rates of FGF8 and FGFR3 in articular cartilage were calculated, and the integrated optical density (IOD) values of FGF8 and FGFR3 positive expression in subchondral marrow were analyzed by Image-Pro Plus 6.0 software. Results:Under light microscope, chondrocytes in low-selenium+ T-2 toxin group were sparse, and empty chondrocytes in the deep and middle layers of articular cartilage increased, and chondrocytes died and became red cell shadows. The extracellular matrix dissolved and was slightly stained in deep region, turning into necrotic and unstructurized areas. Proliferating granulation tissue was visible nearby. The positive expression rate of FGF8 in articular cartilage of rats in low-selenium+ T-2 toxin group [(88.61 ± 10.97)%] was higher than that in control, low-selenium and T-2 toxin groups [(10.35 ± 2.48)%, (19.26 ± 3.08)%, (58.89 ± 9.29)%, P < 0.05]; IOD value of FGF8 positive expression in subchondral marrow [(16.73 ± 1.72) × 10 6] was higher than that in control, low-selenium and T-2 toxin groups [(1.20 ± 0.41) × 10 6, (4.33 ± 0.97) × 10 6, (12.80 ± 1.12) × 10 6, P < 0.05]. The positive expression rate of FGFR3 in articular cartilage of rats in low-selenium+ T-2 toxin group [(89.76 ± 8.59)%] was higher than that in control, low-selenium and T-2 toxin groups [(13.18 ± 2.25)%, (21.15 ± 2.33)%, (32.55 ± 6.72)%, P < 0.05]; IOD value of FGFR3 positive expression in subchondral marrow [(16.50 ± 5.36) × 10 6] was higher than that in control, low-selenium and T-2 toxin groups [(7.58 ± 1.02) × 10 6, (10.73 ± 7.13) × 10 6, (9.83 ± 5.63) × 10 6, P < 0.05]. Conclusion:Under low selenium condition, T-2 toxin changes expression of FGF8 and FGFR3 in deep chondrocytes of articular cartilage and subchondral marrow in rats, elevated expression of FGF8 and FGFR3 may be involved in the occurrence and development of secondary changes in KBD.

Objective:To observe the effects of T-2 toxin on expression of hepatocyte growth factor (HGF) and HGF receptor (C-Met) in articular cartilage and epiphyseal cartilage of rats under low selenium condition.Methods:Twenty-four healthy male SD rats weighted 60-80 g were randomly divided into conventional diet group (selenium content of 101.5 μg/kg) and low-selenium diet group (selenium content of 1.1 μg/kg), with 12 rats in each group. After 30 days of feeding, the conventional diet group was further divided into conventional group and T-2 toxin group (100 μg·kg -1·d -1), and the low-selenium diet group was further divided into low-selenium group and low-selenium+T-2 toxin group (100 μg·kg -1·d -1), with 6 rats in each group. After 30 days of feeding, the rats were sacrificed and the cartilage of knee joint was taken, the morphological changes of knee articular cartilage and epiphyseal cartilage were observed by HE staining under light microscope. Immunohistochemical method was used to detect the expression of HGF and C-Met in knee articular cartilage and epiphyseal cartilage, and positive expression rates of HGF and C-Met were calculated. Results:Under light microscope, chondrocytes of articular cartilage and epiphyseal cartilage in low-selenium+T-2 toxin group were sparse, and the necrosis and structural area were found in the deep layer, and the extracellular matrix of chondrocytes in the region was degraded and light stained, and proliferating granulation tissue was visible nearby. The positive expression rates of HGF in articular cartilage [(21.97 ± 6.90)%, (49.41 ± 8.24)%, (76.39 ± 5.88)%] and epiphyseal cartilage [(23.36 ± 12.49)%, (58.43 ± 14.48)%, (66.59 ± 10.83)%] of rats in low-selenium, T-2 toxin and low-selenium+T-2 toxin groups were higher than those in conventional group [(9.13 ± 6.01)%, (11.14 ± 4.67)%, P < 0.05]. The positive expression rates of C-Met in articular cartilage [(25.34 ± 7.53)%, (58.21 ± 12.54)%, (81.46 ± 7.89)%] and epiphyseal cartilage [(35.21 ± 4.71)%, (40.84 ± 2.03)%, (49.41 ± 6.29)%] of rats in low-selenium, T-2 toxin and low-selenium+T-2 toxin groups were higher than those in conventional group [(11.21 ± 5.11)%, (12.12 ± 4.71)%, P < 0.05]. Conclusion:T-2 toxin may affect the expression of HGF and C-Met in articular cartilage and epiphyseal cartilage of rats under low selenium condition.

Objective To observe the expression level of insulin-like growth factor-1 receptor (IGF-1R) in the cartilage tissue of children with Kaschin-Beck disease (KBD) and T-2 toxin-poisoned rats under low selenium condition,and the effect of IGF-1R inhibitor on apoptosis of human normal chondrocytes (C28/I2 cells),and to investigate the role of IGF-1R in the pathogenesis of KBD.Methods The knuckles of dead children (5 cases) in the KBD areas,car accident death and congenital 6 finger deformity operation children (5 cases) in non-KBD areas in Shaanxi were collected,the expression of IGF-1R in the articular cartilage was detected by immunohistochemistry.Thirty-two male Sprague-Dawley rats with a body mass of 60-80 g were selected,according to the body mass,they were divided into the routine feed group (selenium content:101.5 μg/kg) and the low-selenium feed group (selenium content:1.1 μg/kg) by random number table method,16 rats in each group.After 30 days of feeding,the routine feed group was divided into control group and T-2 toxin group (100 ng·kg-1·d-1),the low-selenium feed group was divided into low selenium group and low selenium + T-2 toxin group,8 rats in each group,the expression of IGF-1R in the articular cartilage of the left knee joint was detected by immunohistochemistry after 30 days of feeding.C28/I2 cells were cultured in vitro and treated with T-2 toxin 0 (control),6,12,and 24 μg/L,and each concentration of T-2 toxin was accompanied with sodium selenite (+ 0.1 mg/L) for 72 h.Meanwhile,IGF-1R inhibitor with 0 (control),250,500,and 1 000 μg/L was treated on C28/I2 cells for 48 h.The expression levels of IGF-1R mRNA and protein in chondrocytes were detected by Real-time PCR and Western blotting,and the apoptosis of chondrocytes was detected by flow cytometry.Results Compared with the control group [(100.00 ± 0.00)％,(100.00 ± 0.00)％],the expression rates of IGF-1R positive cells in articular cartilage surface and middle layers [(72.71 ± 4.75)％,(36.33 ± 4.32)％] of children in KBD group were significantly reduced (t =12.852,32.650,P ＜ 0.01).Compared with control group [(100.00 ± 0.00)％,(100.00 ± 0.00)％,(100.00 ± 0.00)％],the expression rates of IGF-1R positive cells in articular cartilage middle layer [(20.83 ± 2.69)％,(26.45 ± 2.84)％,(20.34 ± 1.82)％],deep layer [(33.55 ± 5.66)％,(48.89 ± 8.39)％,(25.51 ± 7.50)％],and the expression rates of IGF-1R positive cells [(47.50 ± 1.47)％,(28.66 ± 3.58)％,(40.52 ± 6.78)％] in the hypertrophic layer of the metaphyseal plate of rats in low selenium,T-2 toxin,and low selenium + T-2 toxin groups were significantly reduced (P ＜ 0.01).C28/I2 cells were cultured in vitro,compared with the control group,IGF-1R mRNA and protein expression levels in each T-2 toxin groups were significantly reduced (P ＜ 0.05).The expression levels of IGF-1R mRNA (1.95 ± 0.35,2.44 ± 0.17,2.40 ± 0.15) in 6,12,24 μg/L T-2 toxin + 0.1 mg/L selenium groups were significantly higher than those in T-2 toxin groups (0.80 ± 0.08,0.63 ± 0.08,0.61 ± 0.11,t =-12.259,-11.279,-13.371,P＜ 0.05).The expression levels of IGF-1R protein (1.67 ± 0.70,1.07 ± 0.26) in 6,12 μg/L T-2 toxin + 0.1 mg/L selenium groups were significantly higher than those in T-2 toxin groups (0.52 ± 0.05,0.72 ± 0.05,t =-25.977,-10.776,P ＜ 0.05).Compared with the control group [(5.33 ± 0.85)％,(4.03 ± 1.15)％],C28/I2 cells early apoptosis rates [(8.26 ± 1.51)％,(13.00 ± 0.72)％,(13.19 ± 1.05)％] in each of IGF-1R inhibitor groups,and late apoptosis rates [(8.50 ± 0.71)％,(14.21 ± 1.10)％] in 500,1 000 μg/L IGF-1R inhibitor groups were increased significantly (P ＜ 0.05).Conclusions The expressions of IGF-1R in the cartilage tissue of KBD children and T-2 toxin-poisoned rats under low selenium condition are decreased.T-2 toxin decreases the expression of IGF-1R in chondrocytes,and selenium can partly inhibit the effect of T-2 toxin on IGF-1R.Down-regulation of IGF-1R causes chondrocyte apoptosis,and it may play an important role in KBD chondrocyte apoptosis.