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Objective To explore whether tumor necrosis factor receptor-associated protein 1 (TRAP1)gene copy number variation was associated with susceptibility and clinical characteristics of systemic lupus erythematosus (SLE).Methods The study enrolled 304 SLE patients and 391 healthy controls.They were used to investigate the association between TRAP1 gene copy number variation and SLE susceptibility.Then,304 SLE patients were divided into copy number=2 group and copy number＞2 group to study the association between TRAP1 gene copy number variation and disease activity or clinical characteristics of SLE.AccuCopyTM Kit was used to detect the TRAP1 gene copy number.Data analyses were performed by SPSS 10.01 software.The suitable method was selected among t test,rank sum test and x2 test for analysis based on the data type and distribution,univariate and multivariate logistic regression analysis were performed to investigate the associ-ation between TRAP1 gene copy number variation and susceptibility and clinical characteristics of SLE.Results The copy number variation of TRAP1 gene showed an association with the susceptibility to SLE crude OR=5.257,95％CI (1.108,24.937),P=0.037;the adjusted OR=5.578,95％CI (1.172,26.556),P=0.031].There was no association between TRAP1 gene copy number variation and SLE disease activity index (SLEDAI) score (Z=-0.117,P=0.907).The copy number variation of TRAP1 gene had a marginal association with skin lesions in SLE [OR=0.130,95％CI (0.016,1.069),P=0.058],but it disappeared after adjusting for potential confounders [OR=0.288,95％CI (0.029,2.831),P=0.286,PBH=0.808].There was no correlation between TRAP1 gene copy number variation and arthritis,alopecia,oral ulcers,fever,hematologic disorder,lupus nephritis as well as photosensitivity in SLE [x2=0.751,OR=1.234,95％CI (0.767,1.988),P=0.386].No multiplicative interaction was found between TRAP1 gene copy number variation and age or body mass index (BMI) [age:x2=0.751,OR=1.234,95％CI (0.767,1.988),P=0.386;BMI:x2=0.282,OR=1.172,95％CI(0.652,2.109),P=0.596].Conclusions The copy number variation of TRAP1 gene may be associated with susceptibility to SLE.Increased TRAP1 gene copy number may be a risk factor for SLE.
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Objective To detect the effect and mechanism of Cyr61 on the apoptosis of renal tissue caused by early stage of ischemic acute kidney injury (AKI). Methods 30 SD rats were randomized into 5 groups, including control group, AKI group, AKI+bicarbonate group, AKI+blank virus group, and AKI+over?expression Cyr61 virus group. After animal models were created for 2h, serum and renal tissue were collected from sacrificed animals. Expression level of TNF?α was determined by ELISA. HE staining was used to observe the histologic changes of renal tissues. The levels of NF?κB p65 and TNFR1 were measured by immunohistochemical method. RT?PCR and Western blotting assay were adopted to detect the mRNA and protein expression levels of NF?κB p65, TNFR1 and Caspase3. Results Compared with control group, AKI group, AKI+bicarbonate group, AKI+blank virus group, AKI+over?expression Cyr61 virus group had obvious kidney injury. The levels of TNF?α, the mRNA and protein expression levels of NF?κB p65, TNFR1 and caspase3 were markedly up?regulated. Over?expression of Cyr61 significantly attenuated the degree of pathological injury, numbers of apoptotic renal tubular epithelial cells and increased the degree of Scr. Although compared with other groups, the level of TNF?α in kidney tissue had no difference, there was obvious decreased protein level of NF?κB p65, while the increase of TNFR1 and Caspase3 protein was moderate. Conclusions During the early stage of AKI, over expression of Cyr61 could inhibit apoptosis, which may be related to the suppression of TNFR1 transcriptional expression and interference of TNF?αpathway. Its underlying mechanism therefore deserves further research.
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Objective To study the significance of circadian gene Period2 expression in epithelial ovarian cancer tissues and the effect of gene overexpression on the growth of ovarian cancer xenografts in nude mice. Methods Twenty-two cases of ovarian cancer paraffin specimens in the First Hospital of Shanxi Medical University (ovarian cancer group) were chosed during Jau. 2010 to Dec. 2013, including 8 cases of stageⅠ, 8 cases of stageⅡ, and 6 cases of stageⅢ, while 6 cases of benign ovarian epithelial tumor paraffin specimens were selected as control (benign tumor group). Period2 gene were detected by real-time quantitative PCR and western blot methods in different stages of ovarian cancer tumor tissues. Established theovarian cancer xenografts in nude mice with ovarian cancer cell line SKOV3, and they weredivided into 3 groups (n=8), including the recombinant plasmid group, empty plasmid group and control group. Using gene transfection technique to transfer Period2 gene into tumor tissues, tested the expression of Period2 mRNA in tumor tissues by real-time quantitative PCR after transfection into all nude mice, monitoredthetransplant tumor growth and calculating the tumor inhibition rate,detected the antiapoptotic gene BRE, apoptosis related tumor necrosis factor receptor (TNFR1) andtumor suppressor gene NIX in tumor tissues by real-time PCR and western blot in different groups. Results (1)The expression level of Period2 mRNA in tumor tissues amongovarian cancer group stageⅠ,ⅡandⅢwere respectively 2.59±0.50, 0.47± 0.08 and 0.42 ± 0.08, but benign tumor group was 6.59 ± 1.05. The expression level of Period2 protein in ovarian cancer group stage Ⅰ,Ⅱ and Ⅲ were respectively 0.835 ± 0.087, 0.412 ± 0.035 and 0.199 ± 0.031, while benign tumor group was 0.874 ± 0.094. The expression level of Period2 mRNA and protein in benign tumor group was higher than those in ovarian cancer group stageⅠ,ⅡorⅢ(P<0.01). With ovarian cancer stage increased, the expression of Period2 mRNA and protein were decreased or absent (P<0.05).(2)Two weeks after transfection, the expression level of Period2 mRNA in recombinant plasmid group tumor tissue was significantly higher than those in the empty plasmid group or the control group (6.11±0.56 vs 0.50±0.09 vs 0.44 ± 0.08, respectively;P<0.01), the transplanted tumor volume of recombinant plasmid group was significantly less than those in empty plasmid group or the control group[ (486±70) mm3 vs (835±106) mm3 vs (846 ± 110) mm3, respectively;P<0.01], the tumor inhibition rate of the recombinantin plasmid group was as high as 42.9%, that was significantly higher than those in the empty plasmid group and the control group (3.8% and 0, respectively;P<0.05).(3)The expression level of BRE mRNA and protein in transplanted tumor tissues in the recombinant plasmid group were significantly lower than those in empty plasmid group and the control group;the expression level of TNFR1 and NIX were significantly higher than those in the empty plasmid group and the control group (all P<0.05). Conclusions Period2 mRNA and protein expression are absent in ovarian cancer of advanced stage. Transfection and stable expression of Period2 gene could slow down the growth of ovarian cancer, and the tumor inhibition rate could be significantly increased. Period2 gene may promote ovarian cancer cells apoptosis through inhibition of BRE gene expression and promoting TNFR1, NIX gene expressionto exert anti-tumor effect.
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Objective To investigate the effect of tumor necrosis factor receptor 1 (TNFR1) in angiogenesis and neurogenesis during cerebral ischemia in mice.Methods Twenty-four wild-type and 24 TNFR1 knockout mice were randomly divided into either a sham operation group or a focal cerebral ischemia group (n =12 in each group).5-bromodeoxyuridine (BrdU) was injected intraperitoneally at day 3 after cerebral ischemia and sham operation.At day 7 and 28 after cerebral ischemia,the double-label immunofluorescence staining of glucose transporter-1(Glut-1)/BrdU was used to evaluate the angiogenesis surrounding the areas of infarction.A labeled BrdU was used to detect the neural stem cell proliferation in the subventricular zone.Double-labeled doublecortin (DCX)/BrdU and neuronal nuclei antigen (NeuN)/BrdU were used to detect the migration and survival of neural stem cells,respectively.Results Under the normal condition,there was no significant difference in angiogenesis and the number of BrdU-positive cells in the subependymal zone (SVZ) between the wild-type (418.000 ± 28.404) and TNFR1 knockout (528.000 ± 60.597) mice (t =-1.644,P =0.131).At day 7 after cerebral ischemia,the number of Glut-1/BrdU-positive cells in the TNFR1 knockout mice was significantly less than that in the wild-type mice (14.833 ± 2.182 vs.27.5 ± 4.209) (t =2.672,P =0.023),and the number of DCX/B3rdU-positive cells was also significantly less than that in the wild-type mice (163.000 ± 11.106 vs.257.168 ± 12.213) (t =5.705,P =0.000).At day 28 after cerebral ischemia,the number of NeuN/BrdU-positive cells in the TNFR1 knockout mice was significantly less than that in the wildtype mice (6.000 ± 0.577 vs.11.000± 1.571) (t=2.988,P=0.014).Conclusions TNFR1 may play a promoting role in the neurovascular reggeneration in late cerebral ischemia.
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Objeetive:To observe the effect of acupuncture-moxibustion on TNF-a,sTNFR-Ⅰand sTNFR-Ⅱ in rats with Crohn's disease(CD).Method:The models of CD rats induced by TNBS were randomized into model group.herbal cake-partitioned moxibustion group and electroacupuncture group as well as a normal control group.The histopathological changes of colon mucus membrane were observed with HE staining and the contents Of TNF-a.sTNFR-Ⅰand sTNFR-Ⅱ were detected with ELISA method.Results:When compared with the normal group,the TNF-a level in CD rats was substantially elevated and the sTNFR-Ⅰ and sTNFR-Ⅱ showed no marked changes.After herbal cake-partitioned moxibustion and electroacupuncture treatment,the TNF-a level in CD rats was substantially reduced and sTNFRL-Ⅰ and sTNFR-Ⅱ showed no marked changes.The inflammatory lesions and abnormal structures of CD rats were significantly improved after herbal cake-partitioned moxibustion and electric stimulation.Conclusion:Herbal cake-partitioned moxibustion and electroacupuncture Can both remarkably reduce the TNF-a level in blood serum.and this might be one ofthe action mechanisms in the treatment of CD.
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Objective To investigate the characteristic and its clinical value of tumor necrosis factor (TNF)-α and its receptor, interleukin (IL)-1β and its receptor in serum and bronchoalveolar lavage fluid(BALF) in patients with pulmonary tuberculosis and to determine the role of them in the immunopathogenesis of tuberculosis. Methods The concentrations of TNF-α,soluble TNF receptor (sTNF-R) Ⅰ, IL-1β and IL-1 receptor were measured using sandwish ABC-enzyme-linked immunosorbent assay (ELISA) method in serum and BALF of 46 patients with active tuberculosis and 21 patients with inactive tuberculosis, and in the serum of 20 cases of healthy control. Meanwhile the above-mentioned cytokine levels in serum and BALF of 19 patients with active tuberculosis were followed up. Differences between groups were assessed for significance by t test. Results The TNF-α,sTNF-R Ⅰ, IL-1β and IL-1 receptor levels and TNF-α/sTNF-R Ⅰ ratios in BALF of active tuberculosis group were (286.2±96.3) pg/L,(2 431.5±1 124.6) pg/L,(58.6±3.2) pg/L,(162.4±17.1) pg/L and 0.06±0.01, respectively, which were all significantly higher than those with inactive tuberculosis group (t=3.36,3.25,2.95,2.27 and 3.12 respectively; P<0.05). The TNF-α,sTNF-R Ⅰ,IL-1β and IL-1 receptor levels and TNF-α/sTNF-R Ⅰ ratios in BALF of cavernous tuberculosis group were (381.4±106.4) pg/L,(2 824.7±1 318.5) pg/L,(66.4±4.6) pg/L,(176.4±18.7) pg/L and 0.07±0.01, respectively,which were all significantly higher than those of non-cavernous tuberculosis group (t= 3.46,2.37, 3.19, 2.99 and 3.22, respectively; P<0.05). After 2-month' antituberculosis treatments, among 19 cases, the TNF-α,sTNF-R Ⅰ,IL-1β and IL-1 receptor levels and TNF-α/sTNF-R Ⅰ ratios in BALF of 16 cases were significantly lower than those at the beginning of treatments (t= 3.26,3.17, 3.28, 2.92 and 3.12 respectively; P<0.01). Meanwhile, their clinical symptoms improved, sputum smear negative, lesions on chest X-ray resolved and the cavity shrinked or closed. Conclusions TNF-α, sTNF-R Ⅰ, IL-1β and IL-1 receptor are likely to be involved in the immunopathogenesis of tuberculosis. Detection of TNF-α, sTNF-R Ⅰ, IL-1β and IL-1 receptor levels in the serum and BALF is helpful to understand the activity of disease, determine the clinical pattern of disease,assess the prognosis of disease and monitor the therapeutic effect in patients with pulmonary tuberculosis.