Objective To investigate the effect and mechanisms of celecoxib on right heart function in mice with acute high-altitude hypoxia exposure.Methods Male C57BL/6J mice(7 weeks old)were housed in a hypobaric chamber simulating an altitude of 5 800 m for 2 d to establish an animal model of acute hypobaric hypoxia.①Eighteen mice were randomly assigned to plain+saline(P+S),high-altitude hypoxia exposure+saline(H+S),and high-altitude hypoxia exposure+celecoxib(H+Cel).Body weight and routine blood indicators were measured,and cardiac ultrasound examination were performed for heart rate(HR),pulmonary artery acceleration time to ejection time ratio(AT/ET),tricuspid annular plane systolic excursion(TAPSE),tricuspid annular systolic velocity(S'),and left ventricular ejection fraction(LVEF)and fractional shortening(FS).Targeted metabolomic profiling was applied to detect the cardiac arachidonic acid(AA)metabolite levels.The contents of 12,13-dihydroxy-9Z-octadecenoic acid(12,13-diHOME)in the heart,liver,brown adipose tissue,and plasma were quantified by ELISA.② Eighteen mice were randomly assigned into plain+saline(P+S),high-altitude hypoxia exposure+saline(H+S)and high-altitude hypoxia exposure+12,13-diHOME(H+di)groups.Body weight,routine blood tests,and echocardiography were performed as above.③ Thirty-two mice were randomly divided into high-altitude hypoxia exposure+saline(H+S),high-altitude hypoxia exposure+celecoxib(H+Cel),high-altitude hypoxia exposure+soluble epoxide hydrolase inhibitor(sEHI)(H+sEHI),and high-altitude hypoxia exposure+sEHI+celecoxib(H+sEHI+Cel)groups.Body weight,routine blood tests,and echocardiography were performed as above.Cardiac and plasma contents of 12,13-diHOME and epoxyeicosatrienoic acids(EETs)were measured by ELISA.Results ① Compared to the P+S group,the H+S group exhibited significantly reduction of cardiac 12,13-diHOME level(P<0.001),increased counts of white blood cells(WBC)and neutrophils(P<0.01)and decreased TAPSE,S'and AT/ET both at resting state and under stress(P<0.01,P<0.001).Compared to the H+S group,the H+Cel group exhibited significantly increase of cardiac 12,13-diHOME level(P<0.05),reduced WBC and lymphocyte counts(P<0.01,P<0.05)and improved TAPSE and S'levels at resting state and under stress(P<0.01,P<0.001).② Compared to the H+S group,the H+di group demonstrated significantly improvement of TAPSE at basal and under stress(P<0.001)and a trend towards improved TAPSE at resting state(P=0.0532),but no obvious differences was observed in WBC and neutrophil counts between the H+di group and the H+S group.③ Compared to the H+Cel group,both the H+sEHI and H+sEHI+Cel groups exhibited significantly reduction of cardiac 12,13-diHOME level(P<0.01,P<0.05)though no statistical changes in cardiac function indicators.Compared to the H+S group,WBC counts and lymphocyte were decreased,and serum EETs level was incrased in the H+Cel group,H+sEHI group and H+sEHI+Cel group(P<0.01,P<0.001).Conclusion Celecoxib can elevate cardiac level of 12,13-diHOME and improves right heart function in mice after acute high-altitude hypoxia exposure through the CYP450-sEH metabolic pathway.

Objective To investigate whether interleukin-37(IL-37)affects macrophage M1 polarization via nu-cleotide oligomerization of structural domain receptor protein 1(NOD1)/nuclear factor κB(NF-κB)in plasma cell mastitis.Methods A total of 15 patients with plasma cell mastitis were recruited according to the collection standard as inflammatory breast tissue and normal breast tissue with a distance of ≥ 3 cm from the edge of the in-flammatory breast tissue.QPCR was performed to detect NOD1 and IL-3.Phorbol ester(PE)was used to induce THP-1 cells to differentiate into resting macrophages(M0).Lipopolysaccharide(LPS)combined with interferon(IFN-γ)was used to induce the polarization of M0 macrophages towards the M1 phenotype.NOD1 lentiviral RNA interference or over-expression vectors were constructed to regulate the expression of NOD1 in M1 macrophages.M0 macrophages were pretreated with IL-37 recombinant protein and then incubated with LPS and IFN-γ for induction.The expression of NOD1,IL-37,M1 markers(IL-6 and iNOS)and M2 markers(IL-10 and Arg-1)was quantified by qPCR.Western blot was employed to assess the protein level of NOD1,NF-κB p65,and p-NF-κB p65.Co-immunoprecipitation was used to detect the interaction between NOD1 and IL-37.Results Up-regulation of NOD1 and down-regulation of IL-37 as were found in inflammatory breast tissues(P＜0.05).Compared to M0 cells,M1 cells showed up-regulated NOD1 and M1 markers and the elevated phosphorylated NF-κB p65 but the down-regulated IL-37(P＜0.05).In M1 macrophages,both NF-κB inhibitor and NOD1 knockdown led to the down-regulation of NOD1 and M1 markers and caused a decrease in the phosphorylated NF-κB p65(P＜0.05).IL-37 recombinant protein decreased phosphorylation of NOD1,M1 marker and NF-κB p65,which was reversed by over-expression of NOD1.IL-37 may interact with NOD1(P＜0.05).Conclusions IL-37 may inhibit M1 polarization in macrophages by down-regulating NOD1/NF-κB pathway thereby preventing plasma cell mastitis progression.

Aging and age-related ailments have emerged as critical challenges and great burdens within the global contemporary society.Addressing these concerns is an imperative task,with the aims of postponing the aging process and finding effective treatments for age-related degenerative diseases.Recent investigations have highlighted the significant roles of nicotinamide adenine dinucleotide(NAD+)in the realm of anti-aging.It has been empirically evidenced that supplementation with nicotinamide mononucleotide(NMN)can elevate NAD+levels in the body,thereby ameliorating certain age-related degenerative diseases.The principal anti-aging mechanisms of NMN essentially lie in its impact on cellular energy metabolism,inhibition of cell apoptosis,modulation of immune function,and preservation of genomic stability,which collectively contribute to the deferral of the aging process.This paper critically reviews and evaluates existing research on the anti-aging mechanisms of NMN,elucidates the inherent limitations of current research,and proposes novel avenues for anti-aging investigations.

Objective:To investigate the effect of ossotide combined with antibiotics on efficacy and serum levels of tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ) and interleukin-1beta (IL-1β) in patients with brucellosis osteoarthritis.Methods:Using a prospective study, 66 patients with brucellosis osteoarthritis admitted to Jiangshan Beilin Hospital from May 2016 to May 2019 were selected as the study subjects, aged from 28 to 70 years old. The patients were randomly divided into the control group ( n = 33) and the observation group ( n = 33). The control group was treated with antibiotics alone, and the observation group was treated with ossotide injection on the basis of the treatment. Both groups were treated for 2 months. The clinical efficacy was compared between the two groups, the pain visual analogue scale (VAS) scores, the Barthel index of activities of daily living, and the Fugl-Meyer assessment (FMA) scores were compared between the two groups before treatment and 2 months after treatment. The higher the VAS score was, the more severe the pain was. The higher the Barthel index was, the stronger the activity of daily living was. The higher the FMA score was, the better the motor function was. At the same time, the serum levels of TNF-α, IFN-γ and IL-1β were compared between the two groups before treatment and 2 months after treatment. Results:The total effective rate of the observation group (96.97%, 32/33) was significantly higher than that of the control group (78.79%, 26/33), and the difference was statistically significant (χ 2 = 5.12, P < 0.05). After 2 months of treatment, the VAS score [(3.12 ± 0.85) points] of the observation group was significantly lower than that of the control group [(4.08 ± 0.97) points, P < 0.05], Barthel index [(64.72 ± 6.98) points] and FMA score [(58.70 ± 5.58) points] were significantly higher than those of the control group [(56.65 ± 6.72), (51.43 ± 5.33) points, P < 0.05]. After 2 months of treatment, serum levels of TNF-α [(12.30 ± 2.35) ng/L], IFN-γ [(10.87 ± 2.26) ng/L] and IL-1β [(324.28 ± 127.60) ng/L] of the observation group were significantly lower than those of the control group [(14.35 ± 3.13), (15.76 ± 3.21), (390.29 ± 131.37) ng/L, P < 0.05]. Conclusion:Ossotide combined with antibiotics can improve the therapeutic effect of brucellosis osteoarthritis and reduce the serum levels of TNF-α, IFN-γ, and IL-1β.

Background and Objectives: Mesenchymal stem cells (MSCs) have the multipotent capacity to differentiate into multiple tissue lineages as well as to self-renew, which is the main origin of adipocytes. IL6/IL6R pathway exerts a significant role in tissue regeneration and cell differentiation. Whereas, the underlying mechanism between IL6/IL6R pathway and MSCs adipogenesis differentiation remains elusive. Methods: MSCs from healthy donors were cultured in adipogenesis differentiation medium for 0∼14 days, during which their adipogenesis differentiation degree was evaluated by Oil Red O staining. The expression of IL6R was detected in MSCs during adipogenesis differentiation. Knockdown and overexpression of IL6R were respectively performed using siRNA and lentivirus to investigate its effect on MSCs adipogenesis differentiation. The adipogenesis marker genes expression and MAPK pathway activation were detected by Western blotting. The role of P38 pathway in the adipogenesis differentiation of MSCs was determined using the specific inhibitor SB203580. Results: The expression of IL6 and IL6R increased during adipogenesis differentiation in MSCs, which were positively correlated with Oil Red O quantification result. Knockdown and overexpression experiments demonstrated a positive correlation between the expressions of IL6R and MSCs adipogenesis differentiation, accompanied by same trend of P38 phosphorylation. Besides, the specific P38 inhibitor SB203580 markedly inhibited the adipogenesis differentiation potential of MSCs. Conclusions This study reveals IL6R facilitates the adiogenesis differentiation of MSCs via activating P38 pathway.

Ankylosing spondylitis (AS) is a type of autoimmune disease that predominantly affects the spine and sacroiliac joints. However, the pathogenesis of AS remains unclear. Some evidence indicates that infection with bacteria, especially Gram-negative bacteria, may have an important role in the onset and progression of AS. Recently, many studies have demonstrated that mesenchymal stem cells (MSCs) dysfunction may contribute to the pathogenesis of many rheumatic diseases. We previously demonstrated that MSCs from AS patients exhibited markedly enhanced osteogenic differentiation capacity in vitro under non-inflammatory conditions. However, the properties of MSCs from AS patients in an inflammatory environment have never been explored. Lipopolysaccharide (LPS), a proinflammatory substance derived from the outer membrane of Gram-negative bacteria, can alter the status and function of MSCs. However, whether MSCs from AS patients exhibit abnormal responses to LPS stimulation has not been reported. Autophagy is a lysosome-mediated catabolic process that participates in many physiological and pathological processes. The link between autophagy and AS remains largely unknown. The level of autophagy in ASMSCs after LPS stimulation remains to be addressed. In this study, we demonstrated that although the basal level of autophagy did not differ between MSCs from healthy donors (HDMSCs) and ASMSCs, LPS-induced autophagy was weaker in ASMSCs than in HDMSCs. Specifically, increased TRAF4 expression in ASMSCs impaired LPS-induced autophagy, potentially by inhibiting the phosphorylation of Beclin-1. These data may provide further insight into ASMSC dysfunction and the precise mechanism underlying the pathogenesis of AS.
Autoimmune Diseases ; Autophagy* ; Bacteria ; Gram-Negative Bacteria ; Humans ; In Vitro Techniques ; Membranes ; Mesenchymal Stromal Cells* ; Pathologic Processes ; Phosphorylation ; Rheumatic Diseases ; Sacroiliac Joint ; Spine ; Spondylitis, Ankylosing* ; Tissue Donors ; TNF Receptor-Associated Factor 4*

Objective To analyze the influencing factors of strain ratio in diagnosing thyroid lesions of thyroid imaging reporting and data system (TI-RADS) 4-6 categories, and explore the application value of elastic strain ratio in differentiating malignant thyroid nodules from benign ones under different influencing factors.Methods A retrospective analysis was made on the pathologic data of 278 nodules undergone TI-RADS ultrasonography stratification and ultrasound elestography, and 105 nodules of TI-RADS 4-6 categories were selected as research objects. A receiver-operating characteristic (ROC) curve was used to identify the cut-off point for differential diagnosis of thyroid nodules. A logistic regression model was established to analyze the factors affecting strain ratio in the differential diagnosis on thyroid nodules of TI-RADS 4-6 categories. Analyses were made in groups to judging the sensitive, specificity, accuracy, positive predictive value and negative predictive of strain ratio under the impact of different factors. Results The best cut-off point of ROC curve for benign and malignant nodules was 0.54. It was found through the logistic regression model that the strain ratio value was closely related to the size, calcification, and thyroid diffuse disease, and strain ration was of higher value in the thyroid nodules which were less than 1 cm without calcification and thyroid diffuse disease.Conclusions Three factors including size, calcification, and diffuse lesions of the thyroid nodules can influence the elastic strain ratio of TI-RADS 4-6 thyroid nodules. Comprehensive analysis of these three influencing factors can improve the application value of elastic strain ratio in differential diagnosis of benign and malignant thyroid nodules.

BACKGROUND:Ankylosing spondylitis is an autoimmune disease at high inflammatory state, and its pathogenesis is stil unclear. Besides, there is a lack of entirely satisfactory curative strategies. OBJECTIVE: To explore the immunoregulation capability of bone marrow mesenchymal stem cels from ankylosing spondylitis patients on macrophages and the potential therapeutic use of bone marrow mesenchymal stem cels from healthy donors on ankylosing spondylitis. METHODS: Bone marrow mesenchymal stem cels were extracted from 21 healthy donors and 25 ankylosing spondylitis patients respectively, and passage 4 cels were used in subsequent experiments. A human monocytic cel line was induced to differentiate into macrophages. The phenotypic markers of bone marrow mesenchymal stem cels and macrophages were detected by flow cytometry. Expressions of tumor necrosis factor-α and tumor necrosis factor-α-stimulated gene 6 (TSG-6) proteins in the supernatant of co-culture system were detected by ELISA. Quantitative real-time PCR was applied to detect the mRNA level of cytokines secreted by bone marrow mesenchymal stem cels and macrophages. RESULTS AND CONCLUSION:The typical mesenchymal stem cel surface markers were expressed in both bone marrow mesenchymal stem cels from healthy donors and patients with ankylosing spondylitis, and CD68 was detected positively in induced macrophages. The protein and mRNA levels of tumor necrosis factor-α secreted by macrophages co-cultured with bone marrow mesenchymal stem cels from patients with ankylosing spondylitis were obviously higher than those from healthy donors (P < 0.05). TSG-6 secreted by bone marrow mesenchymal stem cels from patients with ankylosing spondylitis was lower than that by bone marrow mesenchymal stem cels from healthy donors in both RNA transcriptional and protein levels (P < 0.05). Our study demonstrates that bone marrow mesenchymal stem cels from patients with ankylosing spondylitis shows abnormal immunoregulatory function on inhibiting the tumor necrosis factor-α secretion from macrophages, which reveals a mechanism of immune disorder in ankylosing spondylitis. The therapeutic mechanism of bone marrow mesenchymal stem cels from healthy donors may work by secreting enough TSG-6 to inhibit the activation of macrophages in patients with ankylosing spondylitis, and thereby to decrease the secretion of tumor necrosis factor-α. Cite this article:Sun SH, Wang P, Su CY, Xie ZY, Li YX, Li D, Wang S, Su HJ, Wu XH, Deng W, Wu YF, Shen HY. Bone marrow mesenchymal stem cels derived from patients with ankylosing spondylitis show abnormal immunoregulation capability on macrophages. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):13-19.

[ ABSTRACT] AIM: To investigate the role of TNF-αand NF-κB in the mechanism of neuropathic pain due to chronically compressed dorsal root ganglion ( CCD) .METHODS:Based on the CCD model, von Frey filaments were used to quantify behavior test.The expression changes of TNF-αand NF-κB were determined by Western blotting, and the cor-relation between the expression of TNF-αand the 50%paw withdrawal threshold was also analyzed.Moreover, the location of TNF-αin dorsal root ganglion ( DRG) was observed with immunofluorescence double staining.RESULTS: We found 50%paw withdrawal threshold of CCD decreased at the first day after operation.The mechanical allodynia was the most ob-vious at postoperative 7~14 d and lasted longer than 35 d.The expression of TNF-αand NF-κB increased significantly in DRG after operation (P<0.01), especially at 7~14 d, and then restored gradually.Moreover, there was a correlation between the protein expression of TNF-αand the changes of neuropathic behavior ( P<0.05 ) .CONCLUSION: TNF-αand NF-κB are involved in the mechanism of mechanical allodynia after chronically compressed DRG.

BACKGROUND:Hip’s anatomy structure is complex, and the traditional two-dimensional imaging, such as X-ray film or CT scan, is susceptible to image overlap and other soft tissue. As a result, there are some limitations in the diagnosis of hip disease. The digital model is built based on reverse engineering and rapid prototyping. It is comprehensive, intuitive and accurate of the stereo configuration of the hip and the spatial relations of other parts’ anatomical structures. This study has a broad application prospect. OBJECTIVE:To reconstruct the personalized three-dimensional entity model of the hip bone with computer aided technology. METHODS: The human hips were scanned with a portable three-dimensional laser scanner, and then the point cloud data of the hips were gotten. The point cloud was processed and the three-dimensional model reconstruction was carried out by using the engineering software Geomagic and the computer aided software CimatronE. Three-dimensional print of hip was completed and the model was gotten, which was consistent with the geometry of hip bone, through the rapid prototyping machine. RESULTS AND CONCLUSION:A model with good geometric similarity and biomechanical characteristics was built by digital modeling method based on reverse engineering and rapid prototyping. This way of digital modeling provided the basis for digital manufacturing of artificial hip, virtual assembly, stress analysis and surgical simulation.