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Ulcerative Colitis (UC) has been reported to be related to Porphyromonas gingivalis (P. gingivalis). Porphyromonas gingivalis peptidylarginine deiminase (PPAD), a virulence factor released by P. gingivalis, is known to induce inflammatory responses. To explore the pathological relationships between PPAD and UC, we used homologous recombination technology to construct a P. gingivalis strain in which the PPAD gene was deleted (Δppad) and a Δppad strain in which the PPAD gene was restored (comΔppad). C57BL/6 mice were orally gavaged with saline, P. gingivalis, Δppad, or comΔppad twice a week for the entire 40 days (days 0-40), and then, UC was induced by dextran sodium sulfate (DSS) solution for 10 days (days 31-40). P. gingivalis and comΔppad exacerbated DDS-induced colitis, which was determined by assessing the parameters of colon length, disease activity index, and histological activity index, but Δppad failed to exacerbate DDS-induced colitis. Flow cytometry and ELISA revealed that compared with Δppad, P. gingivalis, and comΔppad increased T helper 17 (Th17) cell numbers and interleukin (IL)-17 production but decreased regulatory T cells (Tregs) numbers and IL-10 production in the spleens of mice with UC. We also cocultured P. gingivalis, Δppad, or comΔppad with T lymphocytes in vitro and found that P. gingivalis and comΔppad significantly increased Th17 cell numbers and decreased Treg cell numbers. Immunofluorescence staining of colon tissue paraffin sections also confirmed these results. The results suggested that P. gingivalis exacerbated the severity of UC in part via PPAD.
Subject(s)
Animals , Mice , Colitis, Ulcerative/microbiology , Mice, Inbred C57BL , Porphyromonas gingivalis/pathogenicity , Protein-Arginine Deiminases , Virulence FactorsABSTRACT
Objective To evaluate the safety and efficacy of SilverHawk directional atherectomy device in the treatment of femoropopliteal occlusive disease. Methods From August 2012 to June 2014,46 patients(58 limbs)with femoropopliteal occlusive diseases in the treatment by SilverHawk directional atherectomy device were analyzed retrospectively . The mean lesion length and degree of diameter stenosisin the femoropopliteal stenoses(52 limbs) were (4.6 ± 2.3) cm and (85.6 ± 11.3)％.The mean lesion length in the femoropopliteal occlusions(6 limbs)was(6.3 ± 3.2)cm. Rutherford score was 3 ～ 5. Mean ABI was 0.45 ± 0.36. Patency was evaluated with color duplex sonography,CTA and DSA postoperatively. Results 46 patients(58 limbs)were recanalizated suc-cessfully via intraluminal approach. The overall technical success rate was 100％. The procedural success rate was 93.10％. Postoperative residual stenosis and ABI were(10.3 ± 6.2)％and 1.05 ± 0.32,which had statistical diff erence compared with preoperative(t=5.83,P=0.02). The average period of follow-up was 22 months. Mean ABI during the follow-up was 0.96 ± 0.15,which had statistical difference compared with preoperative(t = 5.09,P =0.03). The 6-month and 1-and 2-year primary patency rate was 94.83％、91.38％、84.48％,and secondary patency rate was 98.28％、96.55％、93.10％,respectively. Conclusion SilverHawk directional atherectomy device is safe and effective in treament offemoropopliteal occlusive disease ,with satisfactory early-middle results.
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Objective To study the expression and significance of tumor metastasis suppressor gene-1(TMSG1) in esophageal squamous cell carcinoma (ESCC) and EC109 cells.Methods Immunohistochemistry S-P method was used to examine the expression of TMSG-1 protein in 136 cases of ESCC and 37 cases of normal esophageal mucosa.We analyzed the relationship between TMSG-1 and clinicopathological data of ESCC patients.EC109 cells were treated with 3 μg/mL of cisplatin (CDDP) in vitro for 24 h (the intervention group) and the control group was set up at the same time.The proliferation-inhibitory capability was analyzed with MTT assay.RT-PCR was used to examine the expression of TMSG-1 in the intervention group and the control group.Results The positive rate of TMSG-1 in ESCC and normal esophageal mucosa was 52.2％ (71/136) and 94.6％ (35/37),respectively.The expression of TMSG-1 in ESCC was significantly lower than that in normal esophageal mucosa (P＜0.05).The expression of TMSG-1 was related to TNM stage,differentiation degree and lymph node metastasis (P＜0.05).After EC 109 cells were treated with CDDP for 24 h,the proliferation inhibition rate was increased significantly compared with the control group (P＜0.01).RT-PCR results showed that the expression of TMSG-1 in the cells of the intervention group was significantly higher than that in the control group (P＜ 0.01).Conclusion The abnormal expression of TMSG-1 may play a role in the development and metastasis of ESCC.Examination of TMSG-1 may be useful for making diagnosis and guiding clinical therapy of ESCC.
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Objective@#To explore whether Angiopoietin-like protein 3 (ANGPTL3) is involved in podocyte actin rearrangement, and to analyze whether integrin β3 signal pathway is a key in ANGPTL3 inducing actin rearrangement.@*Methods@#The cultured podocytes were divided into six groups: wild type, ADR treated, ADR+Dex, MOCK, ANGPTL3-cDNA, miRNA, and AD+miRNA group. (1) We observed actin cytoskeleton using Invitrogen reagents with confocal microscopy; (2) Actin cytoskeleton after blocking β3 on podocytes was; (3) The expression of total FAK and p-FAK was through Western blotting.@*Results@#(1) The wild type podocyte's cytoskeleton is arranged orderly. After ADR treatment, podocyte's actin are rearranged and weaken (P<0.05). There was no significant difference in actin arrangement between knock-down and MOCK group. In ANGPTL3-cDNA group the podocyte actin was also significantly rearranged; on the contrary, in miRNA+ADR group, the actin rearrangement never obviously happened (P<0.05). (2) Over-expression of ANGPTL3 podocytes blocked integrin β3 did not happen actin rearrangement. (3) The expression of p-FAK significantly increased in over-expression ANGPTL3 podocytes.@*Conclusion@#ANGPTL3 is a key in inducing actin rearrangement. Intergrin β3 maybe a central pathway in ANGPTL3's role with podocytes.
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Objective To explore whether Angiopoietin-like protein 3 (ANGPTL3) is involved in podocyte actin rearrangement,and to analyze whether integrin β3 signal pathway is a key in ANGPTL3 inducing actin rearrangement.Methods The cultured podocytes were divided into six groups:wild type,ADR treated,ADR+ Dex,MOCK,ANGPTL3-cDNA,miRNA,and AD +miRNA group.(1) We observed actin cytoskeleton using Invitrogen reagents with confocal microscopy;(2) Actin cytoskeleton after blocking β3 on podocytes was;(3) The expression of total FAK and p-FAK was through Western blotting.Results (1) The wild type podocyte's cytoskeleton is arranged orderly.After ADR treatment,podocyte's actin are rearranged and weaken (P ＜ 0.05).There was no significant difference in actin arrangement between knock-down and MOCK group.In ANGPTL3-cDNA group the podocyte actin was also significantly rearranged;on the contrary,in miRNA +ADR group,the actin rearrangement never obviously happened (P ＜ 0.05).(2) Over-expression of ANGPTL3 podocytes blocked integrin β3 did not happen actin rearrangement.(3) The expression of p-FAK significantly increased in over-expression ANGPTL3 podocytes.Conclusion ANGPTL3 is a key in inducing actin rearrangement.Intergrin β3 maybe a central pathway in ANGPTL3's role with podocytes.
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Objective To evaluate the safety and efficacy of SilverHawk directional atherectomy device in the treatment of femoropopliteal occlusive disease. Methods From August 2012 to June 2014,46 patients(58 limbs)with femoropopliteal occlusive diseases in the treatment by SilverHawk directional atherectomy device were analyzed retrospectively . The mean lesion length and degree of diameter stenosisin the femoropopliteal stenoses(52 limbs) were (4.6 ± 2.3) cm and (85.6 ± 11.3)％.The mean lesion length in the femoropopliteal occlusions(6 limbs)was(6.3 ± 3.2)cm. Rutherford score was 3 ～ 5. Mean ABI was 0.45 ± 0.36. Patency was evaluated with color duplex sonography,CTA and DSA postoperatively. Results 46 patients(58 limbs)were recanalizated suc-cessfully via intraluminal approach. The overall technical success rate was 100％. The procedural success rate was 93.10％. Postoperative residual stenosis and ABI were(10.3 ± 6.2)％and 1.05 ± 0.32,which had statistical diff erence compared with preoperative(t=5.83,P=0.02). The average period of follow-up was 22 months. Mean ABI during the follow-up was 0.96 ± 0.15,which had statistical difference compared with preoperative(t = 5.09,P =0.03). The 6-month and 1-and 2-year primary patency rate was 94.83％、91.38％、84.48％,and secondary patency rate was 98.28％、96.55％、93.10％,respectively. Conclusion SilverHawk directional atherectomy device is safe and effective in treament offemoropopliteal occlusive disease ,with satisfactory early-middle results.
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Objective To explore the treatment of multifocal lower extremity arteriosclerosis oblitera-tions. Methods From March 2014 to September 2014, combined procedures were performed on 30 lower limbs in 30 patients with multifocal lower extremity arteriosclerosis obliterations for revascularization. All the patients underwent endovascular , 20 of whom received endarterectomy , 10 received artery emboloctomy , and 8 received profundaplasty. The rates of technical success and clinical success were observed. The patients were followed up for 6-12 months to observe the total patency rate and rate of limb reservation. Results The technical success rate was 100%. The perioperative complication rate was 30% (9/30). 29 limbs gained improvement with differ-ent degree and the clinical success rate was 96.67% (29/30). The ankle-brachial index elevated 0.37 ± 0.19 on average (P < 0.001). Primary patency rate was 90% and 73% at 6 and 12 months, and 12-month limb reserva-tion rate was 97.67%. Conclusions The combined procedures for complex lower extremity arteriosclerosis oblit-erations have a higher short- to mid-term patency rate and limb reservation rate.
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Objective To study TMSG-1 and Cyclin D1 expressions in esophageal squamous cell carcinoma (ESCC) and their relevance to the clinicopathological data and prognosis. Methods Immunohistochemistry S-P method was used to examine the expressions of TMSG-1 and Cyclin D1 in pathological specimens of 136 cases of ESCC and 13 cases of normal esophageal mucosa. Contrast study among immunohistochemistry, clinicopathological data and prognosis was also analyzed. Results (1)The positive expression rates of TMSG-1 and Cyclin D1 in ESCC were 52.2% and 65.4%, respectively. The expressions of TMSG-1 and Cyclin D1 in ESCC were significantly higher than that in normal esophageal mucosa (P<0.05). (2)The expressions of TMSG-1 and Cyclin D1 were all related to clinical stage , differentiation degree and lymph node metastasis (P < 0.05). (3)The expression of TMSG-1 was negatively correlated to the expression of Cyclin D1 in ESCC (r=-0.386,P=0.000). (4)The expression levels of TMSG-1 and Cyclin D1 were independent risk factors in patients with ESCC (P < 0.05). Conclusions The abnormal expressions of TMSG-1 and Cyclin D1 may cooperatively play a role in initiation and development of ESCC. Co-examination of TMSG-1 and Cyclin D1 expressions in primary tumors may be useful for predicting prognosis of ESCC.
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This study was aimed to improve the transfer rate of sinomenine in the intermediate product of Caulis Sinomenii in order to optimize the purification process of intermediate product ofCaulis Sinomenii fromTong-An (TA) injection. The transfer rate of sinomenine and the stability of fingerprints in the intermediate product of Caulis Sinomenii were used as indexes for the investigation on the impact from different pH ofCaulis Sinomenii extract before extraction. The transfer rate of sinomenine was used as an index for the investigation on the impact by different pH of hydrochloric acid created to dry extract solution. The transfer rate of sinomenine was used as an index for the investigation of four separation ways, which included the vacuum filtration, plate and frame filters, high-speed tube separator, and flat direct centrifuge, on the liquid separation of sinomenium acutum acid. The results showed that the pH ofCaulis Sinomenii extract before extraction was 10-11; the transfer rate of sinomenine was the highest in the extraction process and the fingerprints of TA injection was stable. The pH of hydrochloric acid was 2.0-2.5; and the highest transfer rate of sinomenine in acid dissolution process was 92.94%. The high-speed tube separator had the best separation to sinomenium acutum acid-dissolving liquid. The highest transfer rate of sinomenine was 93.34%. It was concluded that the optimized process can effectively improve the transfer rate of sinomenine in the intermediate product ofCaulis Sinomenii. Meanwhile, fingerprints of the product were stable. The process was simple with good repeatability.
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This paper was aimed to study effects of different preparation methods on the content of ginsenosides Rg1,Re and Rb1 in Yi-Xin-Shu (YXS) tablets by HPLC-ELSD.HPLC-ELSD was used as the detection method.The separation and content of ginsenosides Rg1,Re and Rb1 were used as indexes.The influences of three different preparation methods (i.e.,defatted alcohol extraction and butanol extraction,alcohol extraction and butanol extraction,alcohol extraction and butanol extraction ammonia solution washing) on the effect of YXS tablets were studied.Then,the same content determination method was used to compare the influence of alkali washing treatment to ginsenosides Rg1,Re and Rb1 among different batches of Panax ginseng.The results showed that a good separation of ginsenosides Rg1,Re and Rb1 component peak of YXS tablets was achieved by three kinds of separation methods.The separation degree was greater than 1.5.Ammonia solution washing had some effect on ginsenosides Rg1,Re and Rb1 content,which made the content of ginsenosides Rg1,Re and Rb1 be 1.5-1.8 times to those without alkali washing.No effect was shown on the content of ginsenosides Rg1,Re and Rb1 during ammonia solution washing.It was concluded that some other ginsenosides can be transferred into ginsenosides Rg1,Re and Rb1 in YXS tablets solution after ammonia solution washing.
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Objective:To investigate the effects of serum from smoking individuals with periodontitis in the process of Fusobocterium.Nucleatum(Fn)invading KB cells and the expression of matrixmetalloproteinase1(MMP1)of KB cells.Methods:Serum was prepared from 10 smokers with periodontitis(group Y),10 nonsmokers with periodontitis(group N)and 5 periodontal healthy nonsmokers(group H).Serum of 200,400 and 800 μl from the subjects was added to the model of Fn invading KB cells respectively and cultured for 24 hours,the cfu of cellinvaded bacteria was estimated by colony counting.MMP1 protein level in culture supernatant wasmeasured by ELISA.Results:In the 800 μl serum groups,the percentage of invaded cfu was 12.59 ±1.27,8.03 ±0.075 and 7.99±0.14 in group Y,N and H(P ＜0.05)respectively,the concentration(μg/L)of MMP1 in the cultrue supernatant of group Y,Nand H was 400.04 ±21.02,252.57 ±18.89 and 262.47 ±35.29(P ＜0.05)respectively.In the 200 μl and 400 μl serum groups,no significant difference was detected in invasion cfu of Fn in KB cells and MMP1 secrition of KB cells among Y,N and H groups(P>0.05).Conclusion:Higher doses of smokingserum might play a role in the course of Fn invading KB cells and promote the expression of MMP1 secretion from KB cells.
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<p><b>OBJECTIVE</b>To investigate the effects of serum from smoking individuals or non-smoking individuals with periodontitis on Porphyromonas gingivalis (Pg) internalizing KB cells, and the expression of matrix metalloproteinase(MMP)-1, MMP-9, tissue inhibitor of metalloproteinase-1 (TIMP-1) in the culture supernatant of KB cells.</p><p><b>METHODS</b>The venous blood of 20 periodontitis patients' (10 smoking and 10 non-smoking) was extracted under the informed consent and centrifuged for serum. The smoking-individual serum (Y group) and non-smoking-individual (N group) serum were added to the model of Pg internalizing KB cells for 12 hours, plated on brain-heart infusion (BHI) and incubated anaerobically at 37 °C for 5 days. The colony forming units (CFU) of cell-invasive bacteria were estimated by colony counting. MMP-1, MMP-9 and TIMP-1 protein levels in culture supernatant were determined by enzyme-linked immunosorbent assay(ELISA) in the two groups following co-culture of Pg with KB cells for 12 hours.</p><p><b>RESULTS</b>The CFU were (11.2 ± 1.1)×10(4), (12.6 ± 1.2)×10(4), (44.7 ± 1.3)×10(4) CFU/ml when adding 200, 400, 800 µl Y-group serum to the model of Pg co-culture with KB cells and when the serum was extracted from N group, the CFU were (33.6 ± 1.4)×10(4),(38.9 ± 1.1)×10(4), (11.2 ± 1.2)×10(4) CFU/ml respectively. When 200, 400, 800 µl Y group-serum was added to co-culture fluid of Pg internalizing KB cells, the concentrations of MMP-1 secreted from KB cells were (107.2 ± 21.5), (165.9 ± 20.2), (434.4 ± 48.0) µg/L respectively, the concentrations of MMP-9 were (3.99 ± 0.29), (4.21 ± 0.61), (5.62 ± 0.47) µg/L respectively, the concentrations of TIMP-1 were (401.3 ± 12.7), (418.3 ± 28.5), (637.3 ± 37.3) µg/L. When the serum (200, 400, 800 µl) extracted from N group, the concentration of MMP-1 and MMP-9 secreted by KB cell were (77.6 ± 10.8), (84.7 ± 10.2) and (98.2 ± 9.7) µg/L and (3.84 ± 0.52), (4.02 ± 0.68), (4.25 ± 0.37) µg/L, respectively. The concentration of TIMP-1 were (67.3 ± 26.9) , (89.4 ± 22.7) and (78.2 ± 16.5) µg/L secreted by KB cells in the course of Pg internalized KB cell. With the increasing of Y group-serum, the more MMP-1, MMP-9 and TIMP-1 were secreted by KB cells(P < 0.05). When 800 µl Y group-serum was added compared with N group-serum to the Pg co-culture with KB model, the more MMP-1, MMP-9 and TIMP-1 were secreted by KB cells(P < 0.05), when 400 µl Y group-serum was added compared with N group-serum to the Pg co-culture with KB model, the more MMP-1 and TIMP-1 were secreted by KB cells (P < 0.05).</p><p><b>CONCLUSIONS</b>The smoking-serum might enhance Pg internalizing KB cells and enhance the expression of MMP-1, MMP-9 and TIMP-1 secreted from KB cells. The local microenvironment of smoking individual may contribute to the recurrence and progression of chronic periodontitis.</p>
Subject(s)
Humans , Coculture Techniques , KB Cells , Matrix Metalloproteinase 1 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Porphyromonas gingivalis , RNA, Messenger , Serum , Smoking , Tissue Inhibitor of Metalloproteinase-1 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the levels of galectin-3 in the serum and surgical specimens from patients with malignant and benign thyroid lesions and explore their clinical significance.</p><p><b>METHODS</b>Serum samples were collected from 159 patients with thyroid neoplasms and 16 normal subjects for detection of galectin-3 level using ELISA. The expressions of galectin-3 protein and mRNA were also detected in the surgical specimens by immunohistochemistry and in situ hybridization.</p><p><b>RESULTS</b>Galectin-3 protein and mRNA were expressed at a rate of 96.67% in 66 cases of papillary thyroid carcinoma, 83.33% in 6 cases of follicular thyroid carcinoma, 16.67% in 36 cases of papillary hyperplasia, and 11.74% in 51 cases of thyroid adenoma. The expression level of galectin-3 mRNA was slightly lower than but positively correlated with its protein expression. Patients with thyroid carcinoma had significantly higher serum concentration of galectin-3 than those with benign thyroid lesions (papillary hyperplasia and thyroid adenoma) and normal subjects (P<0.001). In patients with papillary thyroid carcinoma, galectin-3 positivity in the tumor tissue was associated with a significantly higher serum galectin-3 level in comparison with the negative cases (P<0.05).</p><p><b>CONCLUSION</b>Detection of galectin-3 in both the serum and surgical specimens can improve the diagnosis rate of thyroid carcinoma.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Case-Control Studies , Early Detection of Cancer , Galectin 3 , Blood , Metabolism , Immunohistochemistry , Thyroid Neoplasms , Blood , Diagnosis , MetabolismABSTRACT
OBJECTIVE@#To investigate the expression and clinical significance of OPN, CD44v6 and MMP-9 in laryngeal squamous cell carcinomas (LSCC).@*METHOD@#The expression of OPN, CD44v6 and MMP-9 in the 47 cases of LSCC and 10 cases normal mucosa were detected by immunohistochemical method. The correlation between OPN, CD44v6 and MMP-9 proteins expressions and the correlation between them and clinicopathological feature were also studied.@*RESULT@#(1) The positive expression rates of OPN, CD44v6 and MMP-9 were 63.8%, 76.6% and 68.1% in LSCC of 47 cases and, and 10%, 30% and 0 in the tumor adjacent issue. (2) The positive rate of OPN and CD44v6 was significantly correlated with the clinical staging, the pathological grading and the metastasis of cervical lymph node (P 0.05). (3) The expression of OPN was well correlated with the positive rate of CD44v6 and MMP-9 in LSCC (r = 0.421, P < 0.01, r = 0.340, P < 0.05). The expression of positive rate of CD44v6 and MMP-9 was well related in LSCC as well (r = 0.376, P < 0.01).@*CONCLUSION@#There is a high level of expression of OPN, CD44v6 and MMP-9 in LSCC. OPN, CD44v6 and MMP-9 are positively related and involved in the invasion and metastasis of LSCC.
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , Metabolism , Pathology , Hyaluronan Receptors , Metabolism , Laryngeal Neoplasms , Metabolism , Pathology , Matrix Metalloproteinase 9 , Metabolism , Neoplasm Metastasis , Osteopontin , Metabolism , PrognosisABSTRACT
Objective To investigate the potential roles of dexamethasone (Dex) in podocyte motility,and to explore the mechanism of modulating α-actinin-4,nephrin.Methods Podocytes were divided into three groups:Dex group [1 μmol/L Dex +50 mg/L puromycin aminonucleoside (PAN)],PAN group (50 mg/L) and normal control group.Scrape wound assay and Transwell migration assay were used to detect cell motility.Filtering ratio of podocytes was measured by FITC labeled BSA.Real-time PCR and Western blotting were used to examine the expressions of c-actinin-4 and nephrin.Results From the scrape wound assays,the ability of wound repair in podocytes of PAN group was significantly increased (P＜0.01),and the number of migrating cells in this group also rose (P＜0.01).Compared to PAN group,podocytes in Dex group did not enhance the motility after treatment with the same dose PAN (P＜0.01).Real-time PCR and Western blotting showed that Dex could significantly inhibit the up-regulated expression of α-actinin-4 and nephrin induced by PAN.Conclusions Dex can relieve the enhanced motility induced by PAN.Its mechanism may be associated with the modulation of the expressions of α-actinin-4 and nephrin.
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AIM: To observe the changes of morphology, the activity of myeloperoxidase (MPO) and membrane pump activities of spleen tissue in acute renal failure (ARF) rabbits, and to inquire into the role of spleen on pathogenesis of immune function disorders during ARF. METHODS: 42 rabbits were divided into control group, HgCl_2 group and glycerinum group. The ARF model was established by hypodermic injection of 1% HgCl_2 at dose of 1.3 mL/kg in HgCl_2 group, intramuscularly injection of 50% glycerinum at dose of 10 mL/kg in glycerinum group, respectively, and the animals were divided into the 12 h, 24 h, 48 h secondary groups (6 rabbits each group). At different time points, the rabbits were cannulated to facilitate the collection of blood sample to examine the biochemical indexes of renal function. The spleen microscopic sections were prepared for observing the morphology. The spleen homogenate was made for determining the activities of MPO and membrane pumping. RESULTS: Pathological sections of spleen showed that the different degree of congestion was found and spleen trabecula was increased in two model groups at multiple-time points. The MPO activity of spleen homogenate in HgCl_2 group and glycerinum group at all time points were obviously higher than that in control group, and at 24 h, the MPO activitie in two model groups was significantly increased than that in the same group at 12 h and 48 h. The activities of Na~+-K~+-ATPase, Ca~(2+)-ATPase, Mg~(2+)-ATPase, Ca~(2+)-Mg~(2+)-ATPase of spleen homogenate in two model groups at multiple time points were significantly lower than those in control group. Following ARF development, the ATPase activitie in two model groups at 48 h was lower than that at 12 h except the Mg~(2+)-ATPase in glycerinum group. CONCLUSION: Spleen as an immune organ has histological damage, arrest of polymorphonuclear neutrophils and dysfunction of membrane pump during the development of ARF in rabbits, leading to immune disorders.