Objective To detect the levels of IL-9,IL-17,and IFN-γ in CD4+T cells from patients with rheumatoid arthritis (RA).Methods The peripheral blood mononuclear cells (PBMCs) from patients with RA and healthy controls were obtained,then the CD4+ T lymphocytes were tested by immunomagnetic beads.The protein levels of IL-9,IFN-γ and IL-17 were measured by flow cytometry (FCM).The mRNA levels of IL-9,IL-17,RORγt and IFN-γwere also detected by qRT-PCR.Data were analyzed by comparison between groups using variance analysis,and Pearson's correlation analysis was used for linear correlation analysis.Results The isolation of untouched human CD4+ T cells from PBMC was effective and its purity was over 90％.The protein levels of IL-9,IL-17,IFN-γwere higher in patients with active RA as compared with patients with inactive RA (P＜0.01) which were (1.62±0.23)％ vs (1.15±0.24)％(P＜0.01),(1.47±0.20)％ vs (1.04±0.26)％(P＜0.01) and (8.1±0.6)％ vs (6.9±1.0)％(P＜0.01) respectively,so did the patients with RA when compared with healthy controls (P＜0.01).The mRNA levels of IL-9,IL-17,RORγt and IFN-γ were higher in patients with active RA as compared with inactive RA patients (P＜0.01),which were (3.0±0.6) vs (1.8±0.4) (p＜0.01) (4.2±0.9)vs (2.3±0.7) (P＜0.01),(4.1±0.7)vs (2.9±0.3) (P＜0.01)and (4.0±0.8)vs (2.3±0.6) (P＜0.01) respectively,so did the patients with RA when compared with healthy controls (P＜0.01).Intracelluar IL-9 levels were positively correlated with IL-17 (r=0.632,P=0.001),IFN-γ (r=0.515,P=0.008),DAS28 (r=0.519,P=0.009) and ESR (r=0.857,P=0.038) but had no correlation with CRP (r=0.38,P=0.61).Conclusion The levels of IL-17,IL-9,IFN-γare higher in the PBMCs of RA patients,and these cytokines may participate in the pathogenesis of RA.

Objective To investigate the venous drainage of lesser saphenous aural neurov enofasciocutaneous distally based flap through fluorescence tracing technique and discuss the pattern of venous drainage.Methods Venous blood for 0.1 ml was collected from every rabbit ear vein of 20 rabbits respectively for separation of the erythrocytes and labeling with FITC.The lesser saphenous sural neurovenofasciocutaneous distally based flaps were successfully established in hind limbs of 20 rabbits that were then allocated into four groups according to different inspection time points at 30 minutes ( Croup A) ,24 hours (Group B) ,72 hours (Group C) and 7 days (Group D) after operation.The labeled erythrocytes (5 μl) were injected into the flaps via lesser saphenous vein (in Groups A and B)or hypoderma (in Groups C and D).Then,the flaps were removed five seconds (in Groups A and B) or 10 seconds (in Groups C and D) after injection,immediately frozen and sectioned (5-7 μm in thickness) for microscopic analysis of fluorescent distribution in the pedicle.Results FITC-labeled red blood cells showed steady green fluorescence under inversion fluorescence microscope.Fluorescence was mainly distributed in the wall of lesser saphenous vein and peripheral vessels,as well as inner and outer membrane of perforator artery.There was only faint fluorescence around sural nerve in Groups B,C and D.HE staining showed that the lumina of lesser saphenous vein in Groups C and D were fully filled with thrombosis.Conclusions Vein of the lesser saphenous sural neurovenofasciocutaneous distally based flaps is refluxed mainly through wall of lesser saphenous vein and peripheral vessels as well as through inner and outer membrane of perforator artery in the pedicle.Thrombosis occurs in the lumina of lesser saphenous but there is no venous blood reflux through the valve of lesser saphenous vein.

Objective To analyze the X-ray,CT and MRI findings of synovial tuberculosis,and to evaluate the role of MRI in diagnosing synovial tuberculosis.Methods Fourteen eases of synovial tuberculosis comfirmed by operation and pathology were retrospectively analyzed and summarized. All patients were examined by MRI and X-ray,and CT scans were performed in 3 cases. Results X-ray showed joint swelling(8 cases),articular space narrowing(7 cases),marginal joint erosions(4 cases), and periarticular osteoporosis(9 cases).The joint swelling was detected on CT in all 3 cases,and bony erosion and speckled sequestra were seen in 2 cases.MRI in all of patients showed joint swelling and synovial proliferation in different drgees,demonstrated as heterogeneously low signal on T_1 WI and slight high signal(7 cases)and obvious high signal(6 cases)on T_2WI,and diffuse synovial proliferation was demonstrated as massive and nodular signal in 8 cases.Joint effusion was present in 7 cases as low signal on T_1WI and high signal on T_2 WI.Osseous erosion lesions were seen in 7 cases,and intra-artieular cartilage thinned,partly or mostly disappeared in 11 cases.Periarticular bone marrow edema was found in 7 cases. Conclusion MRI was superior to X-ray and CT in the diagnosis and differential diagnosis of synovial tuberculosis.

Adenocarcinoma ; metabolism ; pathology ; Chromogranin A ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; methods ; Proliferating Cell Nuclear Antigen ; metabolism ; Staining and Labeling ; methods ; Vimentin ; metabolism

Objective To investigate the immunoregulatory effects of mesenchymal stem cells (MSCs)on peripheral blood T lymphocytes from patients with systemic lupus erythematosus(SLE)in vitro and their potential mechanism.Methods MSCs were isolated from the bone marrow of 3 healthy human volunteers,cultivated and identified.Under phytohemagglutinin(PHA)stimulating,peripheral blood T lymphocytes from 8 patients with SLE were treated with MSCs with the T lymphocyte/MSC ratio being 50:1 in group B and 5:1 in group C or without MSCs(group A).MTT assay was used to detect the proliferation of T lymphocytes.flow cytometry to analyze the expressions of surface markers CD152 and CD28 on T lymphocytes.and real time PCR to measure the mRNA expressions of interleukin-6 and interferon-γ,in the T lymphocytes.Results MSCs could markedly inhibit the proliferation of T lymphocytcs.The proliferation of T lymphocytes expressed as absorbance value at 570 nm was 0.484±0.032 in group B.0.308±0.025 in group C,significantly lower than that in group A(0.765±0.036,both P＜0.05),and significant difference was also observed between group C and B(P＜0.05).In the case of the percentage of CD28 positive T lymphocytes.group B and C were significantly lower than group A(60.39%±3.92%and 45.05%±3.46%vs 74.73%±3.74%,both P＜0.05),and group B significantly differed from group C(P＜0.05).MSCs had no obvious effect on the expression of CD152 on T lymphocytes,but significantly suppressed the mRNA expression of interleukin-6 and interferon-γ(both P＜0.05).and the suppressive effect was enhanced with the incrgase in MSC count.ConclusionsMSCs exert an immunosuppressive effect on T lymphocytes from patients with SLE,likely through inhibiting the proliferation,CD28 expression,interleukin-6 and interferon-γ mRNA expression of T lymphocytes.

Objective To investigate the expression level of CD226 mRNA in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE) and explore the relation between the gene expression and disease activity, and the relation between the gene expression and Gly307Ser polymorphism of CD226 was also examined. Methods CD226 gene was measured with real-time polymerase chain reaction (qRT- PCR) in PBMCs. The expression levels of CD226 gene in PBMCs were compared between 90 SLE patients and 30 healthy individuals. One-way ANOVA and pearson correlation were used for statistical analysis. Results The expression level of CD226 in the PBMCs of SLE patients (6.8±1.1) was significantly decreased compared to healthy individuals (26.5±6.7) (P＜0.01), while there was no association between mRNA level and genotype (P＞0.05). No correlation between ESR, CRP, ANA, SLEDAI scores, C3 and the expression level of CD226 gene was discovered. Conclusion In Hubei Chinese Han population, CD226-Gly307Ser locus is associated with the development of SLE, while T allele does not impact the expression of CD226 gene, thus the role of CD226 gene in autoimmune diseases should be explored in the future.

Objective To investigate the immunoregulatory effects of bone marrow-derived mesenehy-real stem ceils (BMSCs) combined with leflunomide (LEF) on mice T-lymphocytes in vitro. Methods BMSCs from BALB/c mice were isolated and expanded. The purity of BMSCs was identified by flow cytometry (FCM). The BALB/c mice's spleen lymphocytes were isolated by using EZ-Sep<'TM> Mouse 1X. Under ConA stimulation, spleen lymphocytes were pretreated with LEF, then washed and co-cuhured with BMSCs. We set up four groups to investigate in this study: group A, spleen lymphocytes alone; group B, spleen lymphocytes with BM- SCs; group C, LEF-pretreated spleen lymphocytes alone and the group D, LEF-pretreated spleen lymphocytes with BMSCs. T-lymphocytes proliferation was assessed by MTT. FCM was used to analysis T-lymphocytes apoptosis and surface markers of CD69 and CD28. The mRNA expression of interleukin (IL)-2, IL-10 were detected by real-time RT-PCR. Results In vitro, the T-lymphocytes'values of A570 nm were significantly lower in group B and group C, compared with group A (group B vs group C vs group A, 0.578±0.042 vs 0.502± 0.040 vs 0.778±0.035, P<0.01), while the value of A<,570 nm>in group D was 0.218±0.033, which was also obviously lower than that in group B and group C (P<0.01). There were no suppressing effects on T-lympho-cytes'activation and expression of IL-2 had been observed. The proportion of apoptotic T-lymphocytes in group B and group D were (2.29±0.32)％ and (4.22±0.98)％, which was significandy lower than that in group A (8.08±1.20) (P<0.01). The expression of IL-10 in group B and C were also lower than that in group A (group B vs group C vs group A, 0.098±0.039 vs 0.054±0.022 vs 1.000, P<0.01 ). Either, the expression of IL-10 in group D was 0.023±0.015, which was obviously lower than that in group B and group C (P<0.01). Conclusion BMSCs combined with LEF show synergistic immunoregulatary effects on mice's T-lymphoeytes.

Objective To compare the survival rate of two kinds of neurovenofasciocutaneous flaps and investigate the venous reverse flow of flaps.MethodsTen New Zealand White rabbits were randomly allocated into 2 groups of 10 flaps(group A: the lesser saphenous sural pedicled fasciocutanous flaps,blood supply provided with perforator arteries;group B: the lesser saphenous sural pedicled fasciocutanous flaps,blood supply provided with axial type artery).The survival rate of flaps in two groups was observed.Pedicles of flaps were harvested and examined histologically.ResultsThe survival rate of flaps in group A was significantly lower than that in group B[(15.2?16.7)% vs(94.1?6.4)%,P

Objective To investigate the immunoregulatory effects of mesenchymal stem cells (MSCs) on the peripheral T-lymphocytes of lupus nephritis in vitro. Methods MSCs were isolated and expanded from human bone marrow cells. The purity of MSCs was identified by flow cytometry (FCM). The MSCs (4×104, 1×104, 2×103) were added into wells containing peripheral blood lymphocytes (2×105) from lupus nephritis in the presence of phytohemagglutinin [PHA). Samples were divided into the following groups: group A:T-lymphocytes alone; group B: MSCsl with T-lymphocytes(MSCsl:T=1:5); group C: MSCs2 with T-lymphocytes (MSCs2:T=1:20); group D: MSCs3 with T-lymphocytes (MSCs3:T=1:100). The proliferation of T-lymphocytes was assessed by MTT. FCM was used to analyze the apoptosis of T-lymphocytes and surface markers of CD28 and CD152. The gene expression of interferon γ (IFN-γ), interleukin-10 (IL-10), transforming growth factor-β1 (TGF-β1) were detected by real-time RT-PCR. Results In vitro, with the presence of MSCs, peripheral blood T-lymphocytes from lupus nephritis were statistically significantly decr-eased in their proliferative activities , apoptosis and CD28 expression in a dose-dependent manner. No inhibitory effects on CD152 expression of T-lymphocytes had been observed . MSCs promoted the gene expression of TGF-β1 and inhibited the gene expression of IL-10, IFN-γ. Conclusion MSCs can inhibit the proliferative activities, apoptosis and CD28 expression of peripheral blood T-lymphocytes of lupus nephritis, increase gene expression of TGF-β1 and lower the gene expression of IL-10, IFN-γ, which may play an important role in it's immunosuppressive effects on lupus nephritis.

Objective To explore the 3-D anatomical structure of blood vessel around the pancreas and its clinical significance.Methods Fifty objects were scanned by 64-slice CT of GE,the artery,portal vein,spleen vein,superior mesenteric vein and inferior mesenteric vein were 3-D reconstructed by Myrian system,and the scientific data were recorded.Results The artery,portal vein,spleen vein,superior mesenteric vein and inferior mesenteric vein were all reconstructed;the length of portal vein was(44.28±10.23)mm and the length of post-pancreas trunk was(32.13±7.08)mm;the angle between portal vein and spleen vein were classified into three types:the angle of type Ⅰ was<90°,angle of type Ⅱ was=90°,and angle of type Ⅲ was>90 °.30％of the inferior mesenteric vein fed into the spleen vein,56％fell into the superior mesenteric vein and 14％came into the the meeting point of the spleen vein and superior mesenteric vein.Conclusion The 3-D reconstruction of blood vessels around the pancreas can provide the anatomical basis for surgeon and reduce the risk of pancreatic surgery before operation.