Objective： To explore the significance of synovial fluid（SF）anti-cyclic citrullinated peptide (CCP) antibodies  and anti-mutated citrullinated vimentin (MCV) antibodies in the diagnosis of serum negative rheumatoid arthritis (SNRA). Methods: Enzyme linked immunosorbent assay (ELISA) method was apllied in the detection of two groups of patients with knee joint fluid resistance against CCP antibody and antibody of MCV, the experimental group to SNRA patients, a total of 29 cases, and the control for patients with osteoarthritis (OA), a total of 28 cases, and clinical manifestations and laboratory parameters of the two groups were collected. Results: The positive rate of synovial fluid anti-CCP was 34.5% in the SNRA patients, which was significantly higher than 10.7% in the control patients（χ2=4.571, P＜0.05）. The positive rate of synovial fluid anti-MCV was 20.7% in the SNRA patients, which was significantly higher than 7.1% in the control patients（χ2=2.167, P＞0.05）. The SNRA patients of SF anti-CCP and anti-MCV positive had no significant difference from the SNRA patients of SF anti-CCP and anti-MCV negative in age, course and morning stiffness. The levels of erythrocyte sedimentation rate (ESR) , C-reactive protein(CRP) and DAS28 scores in the SF anti-CCP positive patients were higher than those of the SF anti-CCP negative patients. The levels of ESR, CRP and DAS28 scores in the SF anti-MCV positive patients were higher than those of the SF anti-MCV negative patients, (all P<0.01). SF anti-CCP had correlation with ESR, CRP(r=0.567, P<0.01; r=0.664, P<0.01). SF anti-MCV had correlation with ESR, CRP(r=0.344, P<0.01; r=0.749, P<0.01). Conclusion: SF anti-CCP and anti-MCV are helpful for the diagnosis of SNRA and judgement of SNRA activity.

Objective： To detect the anti-citrullinated glucose-6-phosphate isomerase (GPI) 70-88 peptide antibody(anti-C-GPI70-88 antibody),anti-citrullinated GPI 435-453 peptide antibody(anti-C-GPI435-453 antibody), anti-GPI 70-88 peptide antibody(anti-GPI70-88 antibody) and anti-GPI 435-453 peptide antibody(anti-GPI435-453 antibody) in the serum of rheumatoid arthritis(RA) patients,and exa-mine the diagnostic values of the anti-C-GPI peptide antibodies in RA. Methods: The anti-C-GPI70-88 antibody, anti-C-GPI435-453 antibody, anti-GPI70-88 antibody and anti-GPI435-453 antibody were detected by enzyme-linked immunosorbent assay(ELISA) in 191 RA patients,129 other rheumatic diseases and 74 healthy controls. The clinical and laboratory data of the patients with RA were collected, and the values of anti-C-GPI peptide antibodies in the diagnosis of RA and the relationships of anti-C-GPI peptide antibodies with the clinical and laboratory parameters analyzed. Results: (1)The mean titers of the anti-C-GPI70-88 antibody and the anti-C-GPI435-453 antibody in the RA patients (respectively, 68.71±4.20 and 51.78±3.13) were significantly higher than those with other rheumatic diseases and healthy individuals(P<0.05). However,the mean titers of the anti-GPI70-88 antibody and anti-GPI435-453  antibody in the RA patients were similar to those with other rheumatic diseases and healthy individuals.(2)The diagnostic sensitivity and specificity of the anti-C-GPI70-88 antibody for RA were 41.88% and 84.50% respectively; and the diagnostic sensitivity and specificity of the anti-C-GPI435-453 antibody for RA were 46.05% and 86.05% respectively.The sensitivity of combined detection of the two anti-C-GPI peptide antibodies was 50.79%, and the specificity was 81.40%. (3) The positive rates of the anti-C-GPI70-88 antibody and the anti-C-GPI435-453 antibody were 35% and 45% respectively in those patients with negative anticyclic citrullinated peptide antibody, antikeratin antibody, and rheumatoid factor. (4) There was no significant difference in clinical and laboratory indicators between the anti-C-GPI70-88 antibody or anti-C-GPI435-453 antibody positive group and negative group. Conclusion: The anti-C-GPI70-88 antibody and anti-C-GPI435-453  antibody can be detected in the serum of RA patients, and C-GPI may be involved in the pathogenesis of RA. There is a certain diagnostic significance for the sera-negative RA.

Objective： To explore the titer of glucose-6-phosphate isomerase (GPI) for early diagnosis of the outpatient with rheumatoid arthritis (RA) in real life, and to analyze its relationship with disease activity. Methods: In the study, 1 051 patients with arthritis were collected in the group who had joints tender and swelling, and 90 cases of healthy people as a control group. ELISA method was used to detect the serum level of GPI, and according to clinical features and laboratory test, all the patients including 525 RA patients, the other patients including osteoarthritis (OA), 134 cases of seronegative spine joint disease (SpA), 104 cases of systemic lupus erythematosus (SLE), 31 cases of primary Sj-gren syndrome (pSS), 24 cases of gout arthritis (GA), 22 cases of other connective tissue diseases (including polymyalgia rheumatica, dermatomyositis, systemic sclerosis, adult Still disease) and 46 cases of other diseases (including 165 cases of osteoporosis, avascular necrosis of the femoral head, traumatic osteomyelitis, bone and joint disease, juvenile rheumatoid arthritis, tumor). The diagnostic values of GPI were assessed, and the differences between the GPI positive and negative groups of the RA patients in clinical characteristics, disease activity, severity and inflammatory index analyzed. Results: The positive rate of serum GPI in the patients with RA was 55.4%, contrasting to other autoimmune diseases (14.3%) and healthy controls (7.78%)(P<0.001). Compared with the OA and SpA patients, the RA group was increased more significantly, and the difference was statistically significant (P<0.001). The diagnostic value of GPI alone for RA was 0.39 mg/L, the sensitivity was 54.2%, and specificity was 87.3%. The positive rate of GPI in RF negative patients was 36.1%; the positive rate of GPI in anti-CCP antibody negative patients was 34.2%; the positive rate of GPI in RF and anti-CCP antibody negative patients was 24.1%. The level of GPI had positive correlation (P<0.05) with ESR, RF, anti-CCP antibody and HRFIgG. Conclusion: GPI is sensitive in the patients with RA; GPI positive is important in the diagnosis of RA with antiCCP antibody and/or RF negative  patients. The titer of GPI is related with disease activity of RA.

Objective： To detect serum v-raf murine sarcoma viral oncogene homologue B1 (BRAF) protein levels and to investigate their clinical significance in rheumatoid arthritis (RA) patients. Me-thods: Serum samples were obtained from 78 RA patients, 32 osteoarthritis (OA) patients, 16 systemic lupus erythematosus (SLE) patients, 16 gout patients, 16 ankylosing spondylitis (AS) patients, 16 Sj-gren syndrome (SS) patients and 30 healthy controls. BRAF protein in the sera was examined by enzyme-linked immunosorbent assay (ELISA). The associations between BRAF levels and the clinical features including age, sex, disease duration, swelling joints, tenderness joints, duration of moning stiffness, joint deformity, visual assessment scale (VAS) and extra articular manifestations and laboratory parameters including erythrocyte sedimentation rate (ESR), C reactive protein (CRP), rheumatoid factor (RF), disease activity score in 28 joints (DAS28), anti cyclic citrullinated peptide (CCP) antibo-dy, antikeratin antibody, antnuclear antibody (ANA), immunoglobulin and cytokines, such as TNF-α, IL-1β, IL-6 and IL-17A in RA patients were evaluated. Data analyses were performed by using SPSS 19.0 program. Results: The serum BRAF protein levels in the RA patients were significantly higher than those of other rheumatic diseases groups including OA, SLE, AS, SS, gout patients and healthy controls, the P value was 0.002, <0.001, <0.001, <0.001, 0.001 and <0.001 respectively. The level of serum BRAF protein in the RA patients showed a positive correlation with the rheumatoid factor (P=0.009) and IgA levels (P=0.006), but no correlation with clinical features, such as age and duration or other laboratory parameters, including CRP, ESR, antiCCP antibody, IgM, IgG, TNF-α, IL-1β, IL-6 and IL-17A. The RA patients were further divided into normal levels of BRAF protein group and elevated levels of BRAF protein group. Compared with the clinical features and laboratory indexes of normal and elevated levels of BRAF protein groups in the RA patients, there was no significant difference between the two groups in age, duration, DAS28, CRP, ESR, RF, antiCCP, IgA, IgG, IgM, TNF-α or IL-6. Conclusion: The elevated level of BRAF protein in the RA patients showed that BRAF might play a role in the pathogenesis of RA. Further researches on BRAF gene expression may help to clarify the role of BRAF in RA.

Objective： To detect cell frequency and surface markers of peripheral CD4+CXCR5+ follicular helper T (Tfh) cells and analyze the correlation between CD4+CXCR5+Tfh cells and rheumatoid arthritis (RA) disease activity. Methods: Forty RA patients meeting the American College of Rheumatology classification criteria for RA and twenty healthy controls (HC) were included. The peripheral blood mononuclear cells and sera were collected. The expressions of CD4+CXCR5+Tfh cells (CXCR5, C-X-C chemokine receptor type 5) and inducible T cell costimulator (ICOS), programmed death 1 positive (PD-1), interleukin-21 receptor (IL-21R) and CD40 ligand (CD40L) positive on CD4+CXCR5+Tfh cells were analyzed by flow cytometry. The transcript levels of B-cell lymphoma 6 (Bcl-6), as well as IL-21 and IL-21R, were measured by real-time polymerase chain reaction. Besides, serum IL-21 and CXCL13 concentrations were determined by enzyme-linked immunosorbent assay. The potential association between Tfh cells and RA disease activity was detected. Results: The cell surface marker of CXCR5+ on CD4+ cells was significantly increasingly higher across the following groups versus HC: total RA patients (16.75±3.92 vs.7.49±1.84, P＜0.001); RA patients with low disease activity or remission (16.62±3.43 vs. 7.49±1.84, P＜0.001); RA patients with moderate disease activity (16.82±3.07 vs. 7.49±1.84, P＜0.001) and RA patients with high disease activity (16.87±5.50 vs. 7.49±1.84, P＜0.001). Besides, the percentages of ICOS+, PD-1+, IL-21R+ on CD4+CXCR5+Tfh cells in the RA patients were significantly higher than that of HC (ICOS+CD4+CXCR5+cells, 8.37±4.28 vs. 3.72±1.81, P＜0.001; PD-1+CD4+CXCR5+cells, 1.57±1.10 vs. 0.24±0.30, P=0.035; IL-21R+CD4+CXCR5+cells, 4.60±4.05 vs. 0.20±0.19, P=0.006). But the percentage of CD40L+ on CXCR5+CD4+Tfh cells in the RA patients was not significantly higher than that of HC (3.38±3.71 vs. 0.54±0.34, P=0.135). The IL-21R mRNA expression was elevated significantly (5.00±4.94 vs. 0.74±0.55, P<0.001) in the RA patients but not in Bcl-6 mRNA［4.54(3.33, 7.23) vs. 5.31(2.81, 7.44), P=0.329］or IL-21 mRAN［0.72(0.26, 3.45) vs. 0.56(0.27, 3.71), P=0.195］. Additionally, the serum interleukin-21 (IL-21)and CXCL13 levels in the RA patients were higher than in the healthy controls ［IL-21, (200.49±154.56) ng/L vs. (8.21±5.95) ng/L, P<0.001; CXCL13, (93.72±49.72) ng/L vs. (43.09±1.28) ng/L, P<0.001］ and were both positively correlated with RA disease activity indexes, including erythrocyte sedimentation rate, the disease activity score in 28 joints (ESR-based or CRP-based), clinical disease activity index, and simplified disease activity index. However, none of the Tfh cells, anti-citrullinated protein antibody or rheumatoid factor had any relationship with RA disease activity. Conclusion: Peripheral Tfh cells and their relevant cytokines are higher in RA patients than healthy controls, indicating Tfh cells may participate in the pathogenesis of RA, therefore, blocking the pathway of Tfh cells may be reasonable cellular targets for therapeutic intervention.

Objective： To detect the expressions of T follicular helper (Tfh) subsets and T follicular helper effect memory (Tfhem) cells in circulation of patients with rheumatoid arthritis (RA), as well as to examine their roles in providing biomarkers for active RA. Methods: This study enrolled 41 patients with RA, who were na-vely-treated or had no application of hormone and disease-modifying anti-rheumatic drugs in recent 3 months, as well as 32 healthy controls. The percentages of Tfhem (CD4+CXCR5+CCR7lowPD1high) cells, Tfh (CD3+CD4+CXCR5+CD45RA-) subsets, Tfh1 (CXCR3+CCR6-Tfh)，Tfh2 (CXCR3-CCR6-Tfh)，and Tfh17 (CXCR3-CCR6+Tfh), were determined by flow cytometry of peripheral blood from the patients with RA and health controls. Serum levels of cytokines were detected by enzyme-linked immunosorbent (ELISA). The correlations of Tfhem/Tfh subsets with clinical indicators were analyzed. Results: The mean age of the patients was (56.1±14.0) years (range: 20-82 years), the mean disease duration was (8.2±8.1) years. There was no significant difference between the RA patients and the health controls with age and gender. As compared with the health control, the percentage of Tfhem was significantly increased in the peripheral blood of the RA patients (12.8%±5.7% vs. 8.7%±2.0%, P=0.001). Moreover, the increased Tfhem was correlated with the higher disease activity score in 28 joints (DAS28) and erythrocyte sedimentation rate (ESR), but not with other clinical indicators, such as C-reactive protein (CRP), anti-cyclic citrullinated peptide (CCP) antibodies, and rheumatoid factors (RF). In addition, the percentage of Tfh2 subset, but not Tfh1 or Tfh17, was significantly increased in the RA patients (3.002%±0.408% vs. 1.730%±0.160%, P=0.013). As compared with Tfh2-low group, serum levels of Ig (immunoglobulin) A ［(3.045±0.261) g/L vs.(3.963±0.815) g/L, P=0.172］, IgG ［(13.800±0.862) g/L vs.(16.980±0.224) g/L, P=0.161］, IgM ［(1.135±0.083) g/L vs.(1.731±0.380) g/L, P=0.140］, IL (interleukin)-4 ［(2.322±0.214) ng/L vs.(3.994±0.751) ng/L, P=0.056］ and IL-10［(1.898±0.105) ng/L vs. (3.125±0.880) ng/L, P=0.140］ in Tfh2-high group tended to increase with no significant statistical difference. Conclusion: Our data suggest that Tfhem is associated with disease activity and is a va-luable marker for active RA. It also presents a potential pathogenesis in the development of RA and the target for future therapies. Meanwhile, the increased Tfh2 and associated cytokines might be involved in the development of RA.

Objective： The therapeutic potential of umbilical cord mesenchymal stem cells (UC-MSCs) in rheumatoid arthritis (RA) has attracted more and more attention, because of it can suppress the various inflammatory effects of T cells. Galectin-1 is highly expressed in UC-MSCs, as the first lectin mediating the immunomodulatory effect of MSCs. Our study will investigate the effects of galectin-1 in regulation of UC-MSCs on rheumatoid arthritis T cells. Methods: Lentivirus transfected shRNA technique was used to knock down the expression of galectin-1 in UC-MSCs to construct UC-MSCs(Gal-1-). The effects of UC-MSCs and UC-MSCs(Gal-1-) on CD4+ T cells in RA patients were investigated by contact system, including negative control group (CD4+ T cells), positive control group ［CD4+ T-phytohemagg lutinin (PHA)］, UC-MSCs-CD4+ T cells co-culture group, UC-MSCs(control shRNA)-CD4+ T cells co-culture group, and UC-MSCs(Gal-1-)-CD4+ T cells co-culture group. The proliferation of CD4+ T cells was detected by MTS assay. The level of tumor necrosis factors α (TNF-α) in cells supernatant was detected by enzyme linked immunosorbent assay (ELISA). The effect of UC-MSCs on helper T cell (Th) subset was detected by flow cytometry. Results: In vitro, UC-MSCs were capable of inhibiting PHA induced proliferation of CD4+ T cells from RA patients, but UC-MSCs(Gal-1-) did not show the significant inhibitory effect. Galectin-1 affect the TNF-α level of CD4+ T cells regulated by UC-MSCs. UC-MSCs and UC-MSCs(control shRNA) significantly inhibited the expression of TNF-α in PHA-induced CD4+ T cells. However, UC-MSCs(Gal-1-) had no significant inhibitory effect. Furthermore, the Th1 cells were also significantly suppressed by UC-MSCs and UC-MSCs(control shRNA) (4.83%±1.37% and 5.13%±0.87%，P=0.012 and P=0.018). These was no significant difference in the proportion of the Th1 cells between the control group and UC-MSCs(Gal-1-) group (8.51%±2.04% and 6.41%±0.96%，P=0.101). The Th2 cells were protected after silence galectin-1 in UC-MSCs, whereas there was no significant difference. The proportion of Th17 was decreased by co-culture with UC-MSCs and UC-MSCs(control shRNA), but these was also no significant difference. Conclusion: UC-MSCs can inhibit the proliferation and differentiation of CD4+ T cells from RA patients, but these effect declined after knocking down the expression of galectin-1. Galectin-1 maybe take part in the regulation of UC-MSCs on rheumatoid arthritis CD4+ T cells.

Objective： To investigate the effects and mechanisms of adipose-derived stem cells (ADSCs) on bleomycin-induced mice of scleroderma. Methods: In the study, 24 C57BL/6J female mice were randomly divided into  control group, bleomycin（BLM）group, ADSCs (hypodermic injection) group  and  ADSCs (intravenous injection) group . BLM ［2 mg/(kg·d)］ was injected into the mice to establish the model of scleroderma. There were 6 mice in each group .The control group mice were injected with normal saline 2 mL/(kg·d) by subcutaneously. The rest of the three groups were injected with BLM. ADSCs groups were injected with ADSCs (2×105) subcutaneously and intravenously, respectively. T-helper 17 (Th17 ) and regulatory T cell (Treg cell) of spleen cells were detected by flow cytometry. The levels of cytokines in the lung tissue and in the serum were detected by real-time fluorescence quantification. Real-time polymerase chain reaction（PCR） and enzyme-linked immuno sorbent assay（ELISA）. The pathology change of skin and lung tissue was observed by hematoxylin eosin （HE）staining. Results: The proportion of Th17 and Treg increased in BLM group than in control group(15.30%±1.29% vs.4.32%±0.79%; 9.90%±1.95% vs.5.18%±1.35%, P<0.05), the expression of Th17 significantly decreased (5.02%±0.83%, 6.00%±0.82% vs.15.30%±1.29%, P<0.05) and the expression of Treg increased after the ADSCs therapy (14.32%±1.59%, 11.09%±4.31% vs. 9.90%±1.95%, P<0.05). The expression levels of IL-17，IL-6，tumor necrosis factor -α （TNF-α）mRNA in the lung tissue and IL-6 in the serum increased in BLM group than in control group ［3.54±0.30, 10.65±0.66, 5.37±0.52 vs. 1.00±0.00; (21.2±1.74) ng/L vs. (16.87±1.09) ng/L, P<0.05］. The expression of these cytokines significant decreased after the ADSCs therapy ［1.63±0.45,1.50±0.29 vs.3.54±0.30; 3.11±0.85, 2.98±0.76 vs.10.65±0.66;1.45±0.47, 1.59±0.41 vs. 5.37±0.52; (17.87±1.45) ng/L, (17.61±1.16) ng/L vs. (21.2±1.74) ng/L, P<0.05］. But there was no obvious difference between ADSCs (hypodermic injection) group and ADSCs (intravenous injection) group（P>0.05). The expression of TGF-β in the serum increased in BLM group than in control group［(33.95±2.49) ng/L vs. (28.8± 2.29) ng/L, P<0.05］, however, the expression of TGF-β mRNA had no significant differences than that of control group (1.17±0.11 vs.1.00±0.00, P>0.05). The expression of TGF-β mRNA and protein had no significant differences than that of BLM group ［1.25±0.11,1.26±0.12 vs.1.17±0.11; (31.84±2.04) ng/L, (31.25±2.36) ng/L vs. (33.95±2.49) ng/L, P>0.05］. HE staining showed that the inflammation of lung tissue was relieved and the dermal thickness and collagen deposition were decreased after the ADSCs therapy. Conclusion: ADSCs could effectively alleviate inflammation of the lungs and fibrosis of skin; the effects of antiinflammatory and antifibrosis were associated with immune regulating function.

Objective： The ionotropic glutamate receptorantagonists include two types： MK-801， anta-gonist of N-methyl-D-asparticacid (NMDA) receptor, and NBQX, antagonist of non-NMDA receptor.The above-mentioned ionotropic antagonists can block the glutamate and its corresponding receptor binding to produce analgesic effect. The objective of this research was to study two antagonists in analgesic effect on rat behavior，as well as to investigate the down-regulation and up-regulation of cyclooxygenase-2 (COX-2) and Janus-activated kinase (Jak3) in collagen-induced arthritis (CIA) rat serum and tissue fluid after the application of these antagonists, that is, the effect on molecular biology. Methods： This study used the ionotropic glutamate receptors as the target and established CIA rat model. Vivo studies were used to observe changes in behavior and molecular biology of the CIA rat.Behavioral assessment includedmechanical allodynia and joint swelling in the CIA rat，where themechanical allodynia was measured using the paw-withdrawal threshold (PWT) with VonFrey filaments according to the “Up-Down” method，and the drainage volume was used to assess joint swelling. Then the blood samples taken from the heart of the rat and the tissue homogenate were collected to detect the down-regulation and up-regulation of COX-2 and Jak3 in the serum and tissue fluid after the antagonists wereused. Results： Using MK-801, NBQX alone or using the combination of these two antagonists，these three methods all could alleviate pain(P＜0.01).The analgesic effect lasted more than 24 h.Both antagonists reached the peak of analgesia at the end of 4 hours post-injection.NBQX had stronger analgesic effect than MK-801 (P＜0.05).Whether alone or combined use of these two antagonists，could not change the CIA rats’ swelling of the joint (P＞0.05). MK -801 could decrease the expression of COX-2 (P＜0.01).At the same time, NBQX did not have this effect (P＞0.05). Using MK-801, NBQX alone or combination of these two antagonists could not affect the increased expression of Jak3 caused by the CIA (P＞0.05). Conclusion: MK-801 and NBQX could both alleviate pain, NBQX was much better than MK-801. Neither MK-801 nor NBQX had the effect on the swelling of the joint. NMDA receptor and COX-2 inflammatory pathways had certain interactions. For Jak3, it could not be found to have cross-function with ionotropic glutamate signaling pathways by this experiment.

Objective： To analyze the role of anti-collectin 11 in the diagnosis of systemic lupus erythematosus (SLE) and in the evaluation of disease activity. Methods: This was a cross-sectional study. Five groups of patients were enrolled: SLE active (SLE disease activity index-2000，SLEDAI-2000≥9), SLE remission (SLEDAI-2000≤4 and there was no organ involvement), rheumatoid arthritis (RA), primary Sj-gren Syndrome (SS) and healthy control (HC). Serum anticollectin 11 was detected in all the groups by ELISA. One-way ANOVA analysis and LSD-t test as post-hoc analysis were used to compare the levels of anticollectin 11 among all the groups. Receiver operating characteristic (ROC) curve and the area under curve (AUC) were used to analyze the value of anticollectin 11 in the diagnosis of SLE. Results: In the study, 30 patients were enrolled in each group, including 13 males and 137 females with an average age of (34±14) years (18-77 years). The age and gender of the other three groups were comparable to the two SLE groups. The difference of serum anti-collectin 11 between the SLE active group and SLE remission group was not statistically significant (88.8±16.8 vs. 89.7±24.7, P=0.896). The level of serum anti-collectin 11 was significantly higher in SLE group (as a whole) (89.1±19.4) than in RA group (49.1±22.0), SS group (56.9±30.1) and HC group (72.7±24.6) (P＜0.001, P＜0.001, P=0.007, respectively). The AUC was 0.806 for the diagnosis of SLE by serum anti-collectin 11. Further descriptive analysis showed that the positive rate of anti-collectin 11 was very high in the patients of SLE in whom both anti-double-stranded DNA (dsDNA) and Sm antibody were negative. The nervous system and gastrointestinal system involvement were the most common in the patients with positive anti-collectin 11. Conclusion: The level of serum anti-collectin 11 was significantly higher in SLE than in RA, SS and HC. Anti-collectin 11 antibody had a relatively high value in the diagnosis of SLE and it might have some complementary function in the diagnosis of SLE. It might be a relatively specific autoantibody for SLE.

Objective：To construct sphingosine 1-phosphate receptor-1 (S1P1)-small interfering RNA (siRNA) lentiviral vectors and infect human salivary gland cells (HSG), and to investigate its possible therapy on Sj-gren’s syndrome. Methods: HSG cells were divided into blank group, empty vector group, scramble-siRNA group and S1P1-siRNA group. The lentiviral vectors expressing siRNA against S1P1 and the pLL3.7 were respectively transfected into 293T cells with pMD2.G, pMDL g/p RRE, pRSV-REV to produce virus, and then infect HSG cells. The efficiency was observed by flow cytometry after the transfection for 48 h. The expression levels of S1P1 mRNA of HSG were detected by real-time RT-PCR and the expression of S1P1 protein was detected by immunohistochemistry method. The expression levels of interferon-γ (IFN-γ) and interleukin (IL)-17 in the supernatant of the cells were detected by ELISA method. Results:(1) The scramble-siRNA, S1P1-siRNA lentiviral vector was successfully constructed, and the lentivirus titer was about 3.5×108  TU/mL. (2) The level of S1P1 mRNA was lower in S1P1-siRNA group than those in the blank group, empty vector group, and scramble-siRNA group 48 h after infection, there were significant differences between them (P<0.05). (3) The expression of S1P1 protein was lower in S1P1-siRNA group than those in blank group, empty vector group, and scramble-siRNA group 48 h after transfection, there were significant differences between them (P<0.05). (4) The levels of IL-17 were lower in S1P1-siRNA group than those in blank group, empty vector group, and scramble-siRNA group 48 h after transfection, there were significant differences between them (P<0.05). (5) The levels of IFN-γ in S1P1-siRNA group were lower than those in blank group, empty vector group, and scramblesiRNA group 48 h after transfection, there were significant differences between them (P<0.05). Conclusion: The lentiviral vector targeting S1P1 was successfully constructed. S1P1 siRNA could suppress the levels of S1P1 mRNA and protein, and decrease the expression of IL-17 and IFN-γ. S1P1 siRNA could infect HSG cells stably and inhibit the expression of S1P1 gene specifically and efficiently, and reduce the levels of inflammatory cytokines.

Objective： To investigate the association of rs6983267 polymorphism with risk of sporadic colorectal cancer; to compare the distribution of rs6983267 polymorphism between ulcerative colitis and general population. Methods: 186 patients with sporadic colorectal cancer, 129 patients with ulcerative colitis and 189 healthy donors were recruited in the case-control study. Peripheral venous blood was obtained, and genomic DNA was extracted. All samples were genotyped using matrix-assisted laser desorption ionization time-of-flight mass spectrometry techniques. Allelic and genotypic frequencies were compared and adjusted for age and gender using unconditional Logistic regression. Results:  The allelic frequency of G and the genotypic frequencies of GG and GT were predominant in colorectal cancer group compared with control group, which were statistically significant after adjustment for age and gender (P<0.001). The allelic frequency of G and the genotypic frequencies of GG and GT were predominant in ulcerative colitis group compared with control group, which were statistically significant as well (P=0.041, P=0.006 and P<0.001). Conclusion: rs6983267 polymorphism was associated with risk of sporadic colorectal cancer. The distribution of rs6983267 may be different between ulcerative colitis and general population，and the frequency of risk allele G may be higher in ulcerative colitis patients compared with general population.

Objective：To discuss the diagnostic value of carotid atherosclerosis score for ischemic stroke. Methods: In the study, 151 patients with ischemic stroke were enrolled, who were diagnosed by cranial CT scan or cranial MRI scan, and examined with carotid duplex ultrasound, and 151 healthy check-up cases matched by age and sex were chosen as control group, who were excluded ischemic stroke by cranial CT scan or cranial MRI scan. All the control cases were examined with carotid duplex ultrasound also. Intima-media thickness (IMT), the number of carotid plaques, the size of each plaque, the location of the plaque and each plaque’s echo, texture, surface regularity were estimated by carotid duplex ultrasound. Results:  The IMT of the case group and the control group were (0.946±0.185) mm and (0.863±0.148) mm, and there were significant differences (P<0.001); The parameters of arterial plaque correlated with ischemic stroke were plaque’s echo, texture and surface regularity, however the plaque size and location were not correlated with ischemic stroke. The median and quartile of carotid artery plaque score were 3 and 2 respectively in case group, 1 and 2 respectively in control group, and there were significant differences (P<0.001); The parameters of carotid arterial atherosclerosis associated with ischemic stroke were carotid artery plaque score，carotid stenosis degree and IMT, but not the number of carotid plaques. The median and quartile of carotid arterial atherosclerosis score were 5 and 4 respectively in case group, 2 and 4 respectively in control group, and there were significant differences (P<0.001); The area under the curve (AUC) for IMT, the number of carotid plaques, carotid artery plaque score and carotid arterial atherosclerosis score were 0.679, 0.677, 0.704 and 0.805，respectively (P<0.001). The accuracy of carotid atherosclerosis score was the highest. Conclusion: Carotid artery plaque score and carotid atherosclerosis score can be used for the diagnosis of ischemic stroke, and the accuracy of carotid atherosclerosis score is higher.

Objective： To investigate the early complication rate and identify patient-related indepen-dent clinical risk factors for early complications in patients following interventional pulmonology procedures. Methods: In the period from December 2014 to December 2015, sufficient data of Peking University First Hospital Respiratory and Critical Care Medicine Department for analysis were identified in 218 subjects. Interventional pulmonology procedures were performed in all the patients. Early complications after the procedures were defined as newly respiratory failure, arrhythmia requiring treatment, severe hemoptysis, pneumothorax, pneumomediastinum, pulmonary edema, tracheoesophageal fistulae, bronchopleural fistulae, acute coronary syndrome, acute cerebrovascular accident, and death. Patient-related clinical risk factors were defined as coronary atherosclerotic heart disease, cerebral infarction, diabetes mellitus, cirrhosis, chronic kidney disease, arrhythmia, asthma, chronic obstructive pulmonary disease, hypertension, and previous interventional pulmonology treatment. The patient-related independent clinical risk factors which had close relations to the occurrence of early complications were analyzed by multivariate statistical analysis with Logistic regression.  Results: There were 56.4% male and 43.6% female subjects in this study. There were 10.6% current smokers, 26.6% former smokers, and 62.8% non-smokers. The overall early complication rate was 8.3%. In all the subjects groups, the patientrelated independent clinical risk factors for the early complication rate were coronary atherosclerotic heart disease （B=1.545， P=0.006， OR=4.686， 95% CI 1.568－14.006）, chronic obstructive pulmonary disease （B=1.037， P=0.049， OR=2.820， 95% CI 1.675－11.790）, and current smoking status （B=1.412， P=0.032， OR=4.139， 95% CI 1.134－15.109）; for the newly respiratory failure rates were coronary atherosclerotic heart disease （B=2.207， P=0.004， OR=9.087， 95% CI 2.028－40.714）, chronic obstructive pulmonary disease （B=1.646， P=0.048， OR=5.188， 95% CI 1.783－34.375）, and lesions involving three central airways （B=1.899， P=0.032， OR=6.680， 95% CI 1.182－37.740）. In the malignant group, the patientrelated independent clinical risk factor for the early complication rate was current smoking status （B=2.953， P=0.006， OR=19.161， 95% CI 2.360－155.572）. In the benign group, the patientrelated independent clinical risk factor for the early complication rate was only coronary atherosclerotic heart disease （B=1.976， P=0.022， OR=7.214， 95% CI 1.324－39.298）. Conclusion: Closer monitoring of patients with identified clinical risk factors is advisable prior and immediately after interventional pulmonology procedures. In order to avoid or minimize early complications, special attention should be directed toward patients who are current smokers, or patients with lesions involving three central airways, or with coronary atherosclerotic heart disease or chronic obstructive pulmonary disease.

Objective： To analyze the clinical value and prognosis of cesarean scar pregnancy (CSP) treated by uterine artery embolization (UAE). Methods： In the study, 492 cases of patients in Nanjing Maternal and Child Health Care Hospital Affiliated to Nanjing Medical University diagnosed as CSP between January 2011 and December 2014 were chosen, of which 283 were of high-risk group and 209 of low-risk group. According to whether to take UAE, the high-risk group was subdivided into high-risk UAE group(UAE+laparoscopic group), 167 cases, and high-risk non UAE group (chemotherapy+laparoscopic group), 116 cases, while the low-risk group was subdivided into low-risk UAE group (UAE+curettage group), 113 cases, and low-risk non UAE group(chemotherapy+curettage group), 96 cases. The differences of the intraoperative bleeding, length of stay, blood beta human chorionic go-nadotropin (βHCG) dropped to normal time, menstruation recovery time and the hospitalization expenses were compared. And multivariate regression analysis was used to predict the recurrence risk of CSP. Results： The high-risk UAE group was better than the high-risk non UAE group in comparison of intraoperative bleeding ［(36.5±14.8) mL vs.(76.5±39.7) mL］, length of stay ［(5.9±0.9) d vs.(9.6±1.3) d］, blood β-HCG dropped to normal time ［(17.9±8.7) d vs.(28.7±10.1) d］ and menstruation recovery time ［(18.1±1.6) d vs.(24.3±1.8) d］,while the low-risk UAE group was better than the low-risk non UAE group in comparison of intraoperative bleeding ［(93.2±43.3) mL vs.(284.8±110.5) mL］, length of stay ［(10.2±1.4) d vs. (30.7±9.6) d］, blood β-HCG dropped to normal time ［(50.1±17.6)d vs.(67.5±22.9)d］ and menstruation recovery time［(56.3±6.7)d vs.(65.9±9.3) d］, all P<0.05. The highrisk UAE group was higher than the high-risk non UAE group in comparison of hospitalization expenses ［(20 140±1 520 )Yuan vs.(13 510±1 013) Yuan］, and the low-risk group UAE was also higher than the low-risk non UAE group in comparison of hospitalization expenses ［(10 095±962 )Yuan vs.(3 890±457) Yuan］, all P<0.01. Multivariate Logistic regression analysis showed that the treatment method was independent predictor of CSP recurrence risk (OR 2.407, 95%CI 1.176-5.092, P<0.05), and using the comprehensive treatment including UAE could reduce the risk of recurrent CSP. Conclusion： As the efficacy of interventional therapy for CSP was rapid and reliable, fewer complications, faster recovery and lower recurrence, hospitalization with good conditions, and particularly for those patients with CSP who want to fertility again, the comprehensive treatment including UAE treatment should be the first choice.

Objective： To describe the application of polymethylmethacrylate augmentation of bone cement-injectable cannulated pedicle screws for the treatment of degenerative lumbar diseases with osteoporosis. Methods: Observation group included 14 cases of degenerative lumbar diseases with osteoporosis received polymethylmethacrylate augmentation of bone cement-injectable cannulated pedicle screws from November 2014 to July 2015, control group included 12 cases of degenerative lumbar diseases with osteoporosis received polymethylmethacrylate augmentation with traditional pedicle screws.The operation time, blood loss, number of pedicle screws and number of augmented pedicle screws in the two groups were compared. The bone cement leakage and pulmonary bone cement embolism in the two groups were also compared. The fusion rate and pedicle screws loosening by lumbar X ray and dynamic X ray were evaluated. The clinical results were assessed by visual analog scale (VAS) of pain on lumbar and lower limbers, lumbar Japanese Orthopaedic Association scores (JOA), Prolo functional scores and Oswestry disability (ODI) scores. Results: Differences of operation time and blood loss in the two groups were not statistically significant. The average number of pedicle screws was 9.9±4.7 and the average number of augmented pedicle screws was 5.9±2.6 in observation group while the average number of pedicle screws was 7.1±2.8 and the average number of augmented pedicle screws was 3.0±1.9 in control group. The ratio of augmented pedicle screws was higher in observation group than in control group （0.69±0.30 vs.0.47±0.30，P<0.05）. The bone cement leakage rate was lower in observation group than in control group (5/83 vs. 12/42, P<0.01). All the cases in observation group were without leakage to the interspinal canal while one case in control group suffered from bone cement leakage to the interspinal canal with augmentation of 3 pedicle screws. The follow up period was (10.6±2.3) months in observation group and (36.5±7.2) months in control group. In final follow up, no case with nonfusion or pedicle screws loosening was found in both groups. Lumbar VAS, lower limbers VAS, lumbar JOA scores, Prolo functional scores and ODI scores were all better than pre-operation (P<0.01). Conclusion: Polyme-thylmethacrylate augmentation of bone cement-injectable cannulated pedicle screws for the treatment of degenerative lumbar diseases with osteoporosis was effective, with simple working processes and lower risk of bone cement leakage. The short-term clinical result was good.

Objective：To treat the coronal shear fracture of the distal humeral during open reduction and internal fixation by anterolateral approach and lateral approach, and to analyze the advantage and disadvantage of each approach. Methods: From September 2006 to July 2014, 10 patients with coronal fracture of the distal humeral were analyzed, who were all treated with Open Reduction and Internal Fixation (ORIF), 5 with anterolateral approach (group A)  and 5 with lateral approach (group B). For the anterior-lateral approach, the radial nerve and brachioradialis were retracted laterally and the brachialis was retracted medially, the capsule was incised and the fracture line was exposed, usually the capitellum and the lateral part of the trochlear could be exposed clearly but the exposure was limited. For the lateral approach, the brachioradialis was retracted anteriorly, the lateral collateral ligament (LCL) was protected or released from the starting point on the lateral condyle of the humeral, the elbow could be dislocated and the capitellum and part of the trochlear could be exposed. The fractures were classified with the system of Dubberley, the complications were analyzed and the ultimate results were evaluated according to the Mayo elbow performance index (MEPI). Results: For group A, 4 re-operations were performed, 2 for the irritation of the screws,1 for stiff elbow and 1 for failure of the internal fixation. One radial nerve injury happened but recovered later. The mean MEPI was 82 points. For  group B, 1 failure of the internal fixation and instability of the elbow happened. The revision operation was performed for this patient. The mean MEPI was 91 points. Conclusions: Lateral approach is better，it gives more exposure for the joint and the radial nerve is safe, but the trochlear is difficult to be exposed, and the LCL must be protected or repaired during the operation. Anterolateral approach can be used to expose the capitellum and the radial side of the trochlear, but the radial nerve is  dangerous and more complications may happen.

Objective： To identify the preoperative prognostic factors of upper tract urothelial carcinoma (UTUC) and construct preoperative risk stratification system. Methods: A retrospective study including 686 patients who were diagnosed with UTUC and received radical nephroureterectomy or partial ureterectomy in Peking University First Hospital during 2003 and 2013. Results: Of the 686 UTUC patients, 303 (44.2%) were male and 383 (55.8%) female. The postoperative pathological examination showed that 203 (29.6%) had high tumor stages (T3, T4), 300 (43.7%) had high tumor grades (G3) and 54 (7.9%) had lymph nodes metastasis (N1). After multivariate analysis, renal pelvic tumor, large tumor, estimated glomerular filtration rate (eGFR) ≥30 mL/min, and male were associated with high tumor stage. Ureteral tumor, large tumor, and non-smoking history were associated with high tumor grade. Renal pelvis tumor, large tumor, and preoperative anemia were associated with positive N status. During the follow-up, 208 (30.3%) died for cancer and 210 (30.6%) developed intravesical recurrence. Multivariate analysis showed: large tumor (P=0.001), concomitant ipsilateral hydronephrosis (P=0.041), and preoperative anemia (P=0.001) were independently associated cancer-specific mortality after surgery, while ureteral tumor (P=0.04), multiple tumor (P=0.005), and high preoperative creatinine (P=0.036) were independent risk factors for intravesical recurrence. Conclusion: Of the preoperative clinical parameters of UTUC patients, the large tumor, concomitant ipsilateral hydronephrosis, and preoperative anemia were independently associated with cancer-specific mortality after surgery. Ureteral tumor, multiple tumor, and high preoperative creatinine were independently associated with intravesical recurrence after surgery.

Objective： To compare the efficacy and safety of two different shaping methods for double-lumen endotracheal tube (DLT).DLT was shaped with the rod of a Shikani optical stylet (SOS) with the tracheal orifice aligned with the convex aspect of the distal curvature or the concave aspect of the distal curvature. Methods: Patients scheduled for elective thoracic surgery and required intubation with a left-sided DLT were enrolled in this study. They were randomized into two groups. They were intubated with a DLT, which was shaped with the rod of a SOS with its tracheal orifice aligned with the convex aspect of the distal curvature (group T) or the concave aspect of the distal curvature (group U). Time for SOS manipulation, intubation attempts, intubation resistance score, malposition of bronchial intubation, time for fiberoptic bronchoscope (FOB) identification of bronchial placement, total intubation time and oral mucosal or dental injury were recorded. Hoarseness and throat sore of the patients were evaluated 1 hour and 24 hours after surgery. Results: A total of 136 patients completed the study, with 68 in each group. Time for SOS manipulation was significantly shorter in group U ［(35.1±6.1) s vs. 39.6±11.8) s, P=0.007］. First attempt success rate did not differ between the groups (92.6% vs.88.2%, P=0.561). Intubation resistance score was significantly lower in group U. Group T had fewer patients who suffered malposition of bronchial intubation than group U (4 vs.13, P=0.020) and cost less time for FOB identification of bronchial placement ［(44.1±20.9) s vs.(53.6±29.2) s, P=0.032］. Total intubation time and the incidence of oral mucosal or dental injury did not differ between the groups. The severity and incidence of hoarseness were lower in group U than in group T 1 hour after surgery. The severity and incidence of sore throat were lower in group U than in group T 1 hour and 24 hours postoperatively. Conclusion: When lacing a left-sided DLT using a SOS, shaping the DLT with the tracheal orifice aligned with the concave aspect of the distal curvature saves SOS manipulation time, decreases the severity and incidence of postoperative hoarseness and sore throat. However, this modified shaping method increases the incidence of malposition of bronchial intubation and time for FOB identification of bronchial placement.

Objective：The chitosan microspheres encapsulated with bone morphogenetic protein-2 (BMP-2) were prepared by the emulsion cross-linking method. Then the chitosan microspheres were loaded in the ceramic bovine bone (CBB) to achieve the drug delivery system. Methods: The chemical structure and surface morphology of the drug delivery system were investigated by Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscope (SEM) observation. Characterization preserved the loading capacity and encapsulation efficiency of the BMP-2. The dynamic immersion method was used to examine the in vitro release characteristic of BMP-2.  Results: The chitosan microspheres were successfully encapsulated BMP-2 by cross-linking method. The microspheres were micron-sized (5.982 μm) and spherical in shape with smooth surface morphology. From the release experiments, it was found that 5 mg chitosan/BMP-2 soaked for 21 days with a gradual release of BMP-2. The concentration of BMP-2 was (239.1±20.0) mg/L on Day 21. The in vitro experiment showed that this novel drug delivery system could accelerate MC3T3-E1 cells proliferation and osteogenic differentiation. Conclusion: The drug delivery system achieves the biological function of BMP-2 and sustaining slow release in bone repair parts. Also it can provide the basis for repair of bone tissue defect treatment and the selection of bone tissue engineering scaffolds.

Objective：To retrospectively analyze the clinical features, treatment and prognosis to the diffuse tenosynovial giant cell tumor (D-TSGCT) arising from the temporomandibular joint (TMJ), and to give a reference for the early diagnosis and treatment of this disease. Methods: In this study, 15 patients finally diagnosed as D-TSGCT of TMJ histopathologically at the Peking University Hospital of Stomatology from October 2003 to August 2015 were selected and reviewed. Their clinical manifestations, imaging and histological features, diagnoses and differential diagnoses, treatments and follow-ups were summarized and discussed. Results: D-TSGCT of TMJ showed obvious female predominance (12/15), the main symptoms included painful preauricular swelling or mass, limited mouthopening and mandibular deviation with movement. D-TSGCT on computed tomography (CT) scan often showed illdefined soft tissue masses around TMJ, enhancement after contrast administration, usually with widening of the joint spaces and with bone destruction of the condyle, the fossa and even the skull base. On magnetic resonance images (MRI), the majority of lesions on T1 weighted images and T2 weighted images both showed the characteristics of low signals (6/11). The lesions could extend beyond the joints (9/11) and into the infratemporal fossa (4/11) and the middle cranial fossa (4/11). Surgical resection was performed in 14 cases and biopsy in 1 case. Postoperative radiotherapy was performed in 3 cases. In follow-ups, 3 cases showed recurrence postoperatively. Conclusion: D-TSGCT arising from TMJ should be differentiated with TMJ disorders, other tumors and tumor-like lesions of TMJ and parotid neoplasms, etc. CT and MRI examinations have important values in the diagnosis and treatment design of DTSGCT. Because of the local aggressive and extensive behavior, complete resection should be performed as soon as possible. Postoperative radiotherapy was helpful for the extensive lesions including destruction of skull base and may be a good supplementary therapy. Because of the possibility of recurrence and malignancy, long-term follow-up was suggested.

Objective：  To observe the esthetic effect of deep discolored anterior teeth restored by zirconia veneers. Methods: Small defected deep discolored anterior teeth with complete root canal therapy were restored by zirconia veneers (n=15). The same name teeth on the other side of the same dental arch were chosen as control teeth. The color difference values ΔE of the neck 1/3, the middle 1/3 and the incisor 1/3 between the deep discolored tooth and the normal control tooth before and after therapy were measured to evaluate the esthetic effect of zirconia veneer restoration. At the same time, the margi-nal fit of zirconia veneers was checked by the standard of United States Public Health Service (USPHS). The integrity of the veneers was also examined. Results: On the labial side, fibers color difference values ΔE of the neck 1/3, the middle 1/3 and the incisor 1/3 between deep discolored teeth and normal control teeth were measured by the electronic colorimeter, which were 24.92±3.00，26.64±4.00 and 21.94±3.31 respectively. All the values were above 4.0, which were considered unacceptable in clinic. After restoration by zirconia veneers, the color difference values ΔE of the middle 1/3 and the incisor 1/3 between the restored and control teeth were 1.82±0.17 and 1.84±0.21. Both values were less than 2.0, which indicated both good color matching. The color difference value ΔE of the neck 1/3 was 3.92±0.48, which was less than 4.0 and could be accepted in clinic. The statistical analysis of the colors of the teeth before and after restoration compared with the control teeth was done by Paired t test. The t va-lues in the neck 1/3, the middle 1/3 and the incisor 1/3 were 30.37, 21.56, 23.37 respectively. In the three group,  all the P<0.001. There were obvious statistical differences. According to the standard of USPHS, the marginal fit of all the restored teeth was perfect (grade A). No zirconia veneers were broken or detached in the period of observation. Conclusion: Zirconia veneers can be a good method to restore deep discolored anterior teeth. However, it should be used cautiously when the patient’s esthetic expectation was too high.

Objective： To assess the effects of breast-feeding duration, bottle-feeding duration and oral habits on the occlusal characteristics of primary dentition in 3－6-year-old children in Beijing. Methods: This cross sectional study was conducted via an examination of the occlusal characteristics of 734 children combined with a questionnaire completed by their parents/guardians. The examination was performed by a single, previously calibrated examiner and the following variables were evaluated: presence or absence of deep overbite, open bite, anterior cross bite, posterior cross bite, deep overjet, terminal plane relationship of the second primary molar, primary canine relationship, crowding and spacing. Univariate analysis and multiple Logistic regressions were applied to analyze the associations.  Results: It was found that a short duration of breast-feeding (never or ≤6 months) was directly associated with posterior cross bite (OR=3.13, 95%CI=1.11－8.82, P=0.031) and no maxillary space (OR=1.63, 95%CI=1.23－2.98, P=0.038). In children breast-fed for ≤6 months, the probability of developing pacifier-sucking habits was 4 times that for those breast-fed for >6 months (OR=4.21， 95%CI=1.85－9.60， P=0.000 2). The children who were bottle-fed for over 18 months had a 1.45-fold higher risk of nonmesial step occlusion and a 1.43-fold higher risk of class Ⅱ canine relationship compared with those who were bottle-fed for 6-18 months. Non-nutritive sucking habits were also found to affect occlusion: a prolonged digit-sucking habit increased the probability of an anterior open bite, while a pacifier-sucking ha-bit was associated with excessive overjet and absence of lower arch developmental space. Tongue-thrust habit was associated with anterior open bite (OR=4.21, 95%CI=1.85－9.60, P=0.000 2) and posterior cross bite (OR=7.24, 95%CI=1.30－40.13, P=0.024). Lower lip sucking habit was associated with deep overjet and had a negative association with class Ⅲ canine relationship. Unilateral chewing was associated with spacing in mandibular (OR=1.57, 95%CI=1.03－2.41, P=0.037). Mouth breathing was associated with chronic rhinitis and adenoidal hypertrophy and had an association with spacing in maxillary. The chi-square test did not indicate a statistically significant association between upper lip sucking habit and any occlusal characteristics. Conclusion: Breast-feeding duration was shown to be associated with the prevalence of posterior crossbite, or no maxillary space in the deciduous dentition and development of a pacifier-sucking habit. Children who had a longer duration of bottle-fee-ding were more likely to develop class Ⅱ canine relationship. Children who had an oral habit were more likely to develop abnormal occlusal characteristics.

Objective： To prepare felodipine/copovidone solid dispersions, which were made based on different preparation technologies. Insoluble felodipine was selected as the model drug in this research. This drug belonged to Biopharmaceutics Classification System Ⅱ (BCSⅡ) with insoluble property and good permeability across intestinal mucosa simultaneously. A comparative study was carried out for further investigating their corresponding pharmaceutical properties. Methods: Felodipine/copovidone solid dispersions were achieved by four methods including spray-drying method, microwave-induced fusion quench cooling method, freeze-drying method and co-precipitation method. These solid dispersions were produced based on corresponding processes that corresponded to these methods. Internal properties of co-povidone solid dispersions were analyzed by various approaches including scanning electron microscope (SEM), differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD). The improvement on insoluble properties of felodipine by solid dispersions produced by different technologies was characterized by dissolution experiments based on dissolution instrument. Crystallization inhibition effect of polymers against drugs was studied by supersaturated experiments through determining the concentration value at different time points. Results: The internal drug was dispersed in amorphous form in solid dispersions produced by spray-drying, microwave method, microwave/quench-cooling method and co-precipitation method. Freeze-drying method resulted in a form of crystal in felodipine/copovidone solid dispersions. Compared with other technologies, microwave-induced quench cooling method could significantly improve the dissolution of insoluble drug felodipine (P<0.05). The dissolution concentration reached approximately 4.65 mg/L at 60 min time point. Copovidone could inhibit or retard the crystallization of felodipine in a supersaturated state. In the solution pre-dissolved with maximum copoyidone po-lymer, the minimum crystallization rate of supersaturated felodipine was observed at 240 min time point. The value of crystallization rate was 0.19 mg/(L·min). Conclusion: The study is helpful to understand and clarify the internal properties of solid dispersions obtained by different technologies. The research also provides beneficial consultation for the choice of technology in practical production of drug-polymer solid dispersions.

IgG4-related disease is a systemic disorder involving a spectrum of multiple indications, and various histopathological features are shared among different IgG4-related disease subtypes, which challenge diagnosis, although certain syndromes have organ-specific involvement. Among them, Mikulicz’s disease affecting the salivary and lacrimal glands, distinguished by often elevated levels of serum IgG4, infiltration of IgG4+ plasma cells into target tissues, and diffuse swelling, mass formation, or fibrosis of affected organs. However, there are several diseases, which could manifest as salivary gland swelling, mimicking Mikulicz’s disease, such as Sj-gren’s syndrome, mumps virus infection, obstruction of parotid duct, non-Hodgkin’s lymphoma (NHL), and so on. So differential diagnosis is important and essential as to the salivary gland swelling. In this paper, we analyzed a case of a 59-year-old male with symmetric salivary gland swelling. Mikulicz’s disease was misdiagnosed at the beginning without biopsy. Prednisone treatment ever seemed to be effective and antibiotics had no effect. Besides salivary involvement, the patient also manifested as testicle swelling and severe pancytopenia with the development of the disease, which rarely appeared in Mikulicz’s disease. Physical examination showed skin, sclera yellow dye, swollen submandibular, sublingual and lacrimal gland and splenomegaly. As a result, biopsy of right submandibular gland was made, and mucosa-associated lymphoid tissue lymphoma was confirmed by morphology and immunohistochemistry. Bone marrow biopsy also confirmed that lymphoma cells were found in the bone marrow. Finally, the diagnosis of mucosa-associated lymphoid tissue lymphoma (Phase ⅣE, Group A) was made on the patient, who was transferred to the hematology department for the treatment. NHL, especially, primary extranodal lymphoma usually involves the salivary gland, and painless swelling of the salivary gland is a common manifestation, similar with Mikulicz’s disease. So although salivary gland swelling is often associated with autoimmune diseases such as Sj-gren’s syndrome and IgG4-related disease, the awareness and suspicion of a possibility of NHL are essential for rheumatologists. Biopsy is a necessary examination to decrease or avoid misdiagnosis.

Sj-gren’s syndrome (SS) is a chronic autoimmune disorder characterized by lymphocytes infiltration in the exocrine glands. Central nervous system complications of primary SS are not rare, but ischemic stroke has been rarely reported. Here we report a 43-year-old female with a two-year history of primary SS, presenting with sudden cerebral infarction. Her primary SS was diagnosed on the basis of clinical features, high levels of serum anti-SSA and anti-SSB antibodies, salivary gland secretion evaluation and positive sublingual gland biopsy results. Magnetic resonance imaging revealed infarct lesions in the parietal and occipital lobes, as well as in the left basal ganglia. Magnetic resonance angiography showed a remarkable stenosis in the left middle cerebral artery. Other differential diagnoses were ruled out. Corticosteroid and immunosuppressor, together with anti-platelet and statin were effective, and the patient recovered quickly without sequelae. Based on these findings, vasculitis due to primary SS should be considered among the causes of stroke. The literature was reviewed and the relationship between primary SS and cerebral infarction explored. The pathogenesis of ischemic stroke in primary SS is still unknown and warrants further studies.

Episcleritis and scleritis are relatively rare ocular diseases, which are commonly associated with rheumatic diseases including systemic lupus erythematosus (SLE). To investigate clinical and laboratory features of SLEassociated episcleritis and scleritis, we now report 4 cases of inpatients who were diagnosed with episcleritis or scleritis secondary to SLE from September 2005 to July 2016 in the Department of Rheumatology and Immunology in Peking University People’s Hospital. Demographic, clinical and laboratory characteristics were summarized together with the treatment regimen and the prognosis; the literature was reviewed. There were 3 female and 1 male patients. The average age was (49.0±23.8) years and the mean duration of SLE at the onset of episcleritis or scleritis was (2.1±1.4) years. In addition to the eye involvement, the patients had mucocutaneous manifestations, serositis, lupus nephritis and interstitial pneumonia simultaneously;  in the past, 1 patient experienced arthritis, 2 presented Raynaud’s phenomenon, and 2 had hematologic involvement. All the patients had antinuclear antibody (ANA) of high titer. The anti double-stranded DNA (ds-DNA) antibody titers were increased in 2 patients. Three patients had positive anti-nucleosome antibody (ANuA) while the other 1 patient did not test it. The complement levels were decreased in 3 patients. The systemic lupus erythematosus disease activity index (SLEDAI) scores were more than 4 points in all the patients (ranging from 7-16), suggesting active disease. Ocular symptoms included pain, redness of the eye and tears. Ophthalmic examinations revealed 3 cases of episcleritis and 1 case of scleritis. Among the 4 patients, 2 patients expe-rienced ocular complications including decrease in vision and uveitis. All the patients were treated with systemic corticosteroids combined with hydroxycloroquine; 3 patients were treated with immunosuppres-sants (cyclophosphamide in 2 patients and leflunomide in 1 patient). All of the 4 patients received topical steroid and 1 patient received periocular injection of triamcinolone acetonide; 1 patient received topical nonsteroidal anti-inflammatory drug (NSAID).No recurrence of episcleritis or scleritis was observed during the follow-ups. As a conclusion, scleritis and episcleritis, although uncommon, may occur in patients with autoimmune rheumatic diseases including SLE. The occurrence of episcleritis and scleritis may suggest active disease of SLE. Ocular complications need to be aware of in the patients. Prompt diagnosis and treatment was associated with good visual outcomes in the follow-ups.

DiGeorge syndrome is the most common chromosome microdeletion disease. The classical complications include congenital heart disease, hypothyroidism, immunodeficiency, facial abnormalities, and hypocalcemia. According to whether there is an absence or hypoplasia of the thymus, DiGeorge syndrome can be divided into two types, complete DiGeorge syndrome and partial DiGeorge syndrome. The patient was a female born with congenital heart disease, facial abnormalities and cleft palate. When the patient went to school, she had learning difficulty and had problems in communication and personal social behavior. Breath-holding occurred when she was 6 years old. She got infections about 2-3 times a year, which was easy to be cured each time. Chromosome microdeletion test of peripheral blood showed the classical 22q11.2 microdeletion, and no evidence showed that she has thymus absence, thus her disease was diagnosed as partial DiGeorge syndrome. When the patient was 6 years old, the blood routine test showed slight thrombocytopenia, and reexaminations after that indicated the similar result. When 9 years old, she was found with anemia and severe thrombocytopenia. At the age of 10, the patient was admitted to our hospital, complaining of petechia in the body and mucous of mouth. According to the various examinations results, doctors eventually considered the situation as an autoimmune disorder phenomenon. After being treated by pulse-dose methylprednisolone for three days, the bleeding ceased. Then the patient orally took prednisone acetate and pulse-dose cyclophosphamide, however the thrombocyte and hemoglobin levels had not been back to a normal range. But when the dose of prednisone acetate was reduced, the blood platelet count declined again while the hemoglobin kept normal. The long-term follow-up of this case lasted for more than 20 years. Until now, the patient is taking orally prednisone acetate as a maintainance treatment, and the anemia has been improved since, but thrombocytopenia still exists. The mechanism of DiGeorge syndrome in combination with immunodeficiency is still unclear. The most likely reason is that this phenomenon has some relationship with the dysfunction of the thymus and finally had an effect on the function of T cells. The clinical manifestation is always stubborn and need treatment and follow-up visit for a long time

For ideal implant rehabilitation, an adequate bone volume, optical implant position, and stable and healthy soft tissue are required. The reduction of alveolar bone and changes in its morphology subsequent to tooth extraction will result in insufficient amount of bone and adversely affect the ability to optimally place dental implants in edentulous sites. Preservation of alveolar bone volume through ridge preservation has been demonstrated to reduce the vertical and horizontal contraction of the alveolar bone crest after tooth extraction and reduce the need for additional bone augmentation procedures during implant placement. In this case, a patient presented with a mandible molar of severe periodontal disease, the tooth was removed as atraumatically as possible and the graft material of Bio-Oss was loosely placed in the alveolar socket without condensation and covered with Bio-Gide to reconstruct the defects of the alveolar ridge. Six months later, there were sufficient height and width of the alveolar ridge for the dental implant, avoiding the need of additional bone augmentation and reducing the complexity and unpredictability of the implant surgery. Soft tissue defects, such as gingival and connective tissue, played crucial roles in long-term implant success. Peri-implant plastic surgery facilitated development of healthy peri-implant structure able to withstand occlusal forces and muco-gingival stress. Six months after the implant surgery, the keratinized gingiva was absent in the buccal of the implant and the vestibular groove was a little shallow. The free gingival graft technique was used to solve the vestibulum oris groove supersulcus and the absence of keratinized gingiva around the implant. The deepening of vestibular groove and broadening of keratinized gingiva were conducive to the long-term health and stability of the tissue surrounding the implant. Implant installation and prosthetic restoration showed favorable outcome after six months.

Rheumatoid arthritis (RA) is a destructive chronic autoimmune disease characterized by synovium inflammation, cartilage destruction, bone erosion and the presence of autoantibodies. Hypoxia is a prominent micro-environmental feature in a range of disorders including RA. A combination of increased oxygen consumptionby inflamed resident cells and infiltrating immune cells along with a disrupted blood supply due to vascular dysfunction contribute to tissue hypoxia in RA. Hypoxia in turn regulates a number of key signaling pathways that help adaptation. The primary signaling pathway activated by hypoxia is the hypoxiainducible factor (HIF) pathway. It has been shown that HIFs are highly expressed in the synovium of RA. HIFs mediate the pathogenesis of RA through inducing inflammation, angiogenesis, cell migration, and cartilage destruction, and inhibiting the apoptosis of synovial cells and inflammatory cells. HIF expressed in RA can be regulated in both oxygendependent and independent fashions, like inflammatory cytokines, leading to the aggravation of this disease. Considering the vital role of HIF in the pathogenesis of RA, we reviewed the new advances about hypoxia and RA. In this review, we firstly discussed the hypoxia-inducible factor and its regulation, and then, the pathologic role of hypoxia in RA, mainly elucidating the role of hypoxia in synovitis and cartilage destruction and immune cells. Finally, we provided evidence about the potential therapeutic target for treating RA.

Systemic lupus erythematosus (SLE) is a prototypic systemic autoimmune disease, which characterized by complex immunological abnormalities and multiple tissue and organ damages. The etiology and pathogenesis of SLE have not been entirely recognized. Genetic, environmental and viral infections and other factors might be related to the pathogenetic mechanisms of SLE. Interleukin-2 (IL-2) is a critical cytokine produced by T cells upon activation and is important for the generation of T regulatory cells and activation-induced cell death. In SLE patients, T cells display decreased capacity to produce IL-2. Impaired IL-2 expression resulted in decreased generation of regulatory T lymphocytes, and defect of activation-induced cell death. Former researches indicated that IL-2 deficiency in SLE is important for the pathogenesis and treatment of SLE. Several regulating molecules can affect the transcription of IL-2 gene and had an important role in the pathogenesis of SLE. These molecules include cyclic AMP-responsive element modulator (CREM), protein phosphatase 2A (PP2A), E-74 like factor 1 (Elf-1), B lymphocyte induced maturation protein-1 (Blimp-1) and interferon regulator factor 5 (IRF-5). CREM is a transcriptional inhibitor that can repress the transcription of the IL-2 gene by binding to the promoter of the IL-2 gene. PP2A is a Ser/Thr phosphatase that expressed in eukaryotic cells ubiquitously, it represents a negative regulator of the IL-2 gene promoter activity. Elf-1 belongs to the Ets family of transcription factors and can promote the expression of IL-2. Blimp-1 is a crucial transcription factors for regulating B lymphocyte terminal differentiation, an important function of Blimp-1 in T cells is to repress IL-2 gene transcription directly. Interferon regulatory factors (IRFs) are distinctive transcriptional regulators of type Ⅰ interferons (IFNs) and IFN inducible genes, IRF-5 is a member of the IRFs family. IRF-5 is found to be increased in SLE and can regulate the production of IL-2 negatively. PP2A can inhibit the synthesis of IL-2 in two ways: on the one hand, activating the IL-2 transcription inhibitory factor CREMα, on the other hand, inhibiting IL-2 stimulating transcription factor Elf-1. While IRF-5 can activate the IL-2 transcription negative regulator Blimp-1 as to inhibit IL-2 expression. These molecules participate in the regulation of IL-2 through different pathways. This paper reviews the current knowledge of IL-2 signaling pathway regulating molecules in SLE.