OBJECTIVE:To investigate the effect of signals mediated by activated CD28 in promoting survival of multiple myeloma (MM) cells and metabolic fitness and its possible mechanism. METHODS:The expression of CD28 on 4 MM cell lines (XG2, XG1, RPMI 8226 and U266) was determined by flow cytometry. Two cell lines with the highest or lowest CD28 expression were selected. The proliferation, cell cycle, migration and apoptosis of MM cells in vitro were determined in medium containing high glucose concentration or CD28 agonist monoclonal antibody with different bioassays. shRNA interference assay was used to knock down the expression of CD28 on U266 cells. Then, the effect of activated CD28 on glucose uptake rate and drug resistance in MM cells were analyzed using fluorescent glucose analogues (2-NBDG). The expression of Glut1/4, HkII and Fasn was determined with real time quantitative PCR. RESULTS:Flow cytometry analysis showed that all the four tested MM cell lines expressed CD28 and U266 cells had the highest positive rate. The results of in vitro experiment showed that CD28 activation could significantly up-regulate the expression of Glut4 and HkII, promote MM cell metabolic remodeling, enhance 2-NBDG/glucose uptake, increase energy metabolism, thereby elevating cell proliferation and migration abilities, leading to an increase in the number of cells in S- and G2-phases. Meanwhile, activated CD28 subsequently up-regulated resistance of MM cells to bortezomib or dexamethasone. CONCLUSION:MM cells express high levels of CD28 abnormally, and activation of CD28 can promote up-regulation of glucose uptake in MM cells, thereby promoting cell proliferation and enhancing drug resistance.

OBJECTIVE:To investigate the effect of adipocytes in the bone marrow microenvironment of patients with multiple myeloma (MM) on the pathogenesis of MM.METHODS:Bone marrow adipocytes (BMA) in bone marrow smears of health donors (HD) and newly diagnosed MM (ND-MM) patients were evaluated with oil red O staining. The mesenchymal stem cells (MSC) from HD and ND-MM patients were isolated, and in vitro co-culture assay was used to explore the effects of MM cells on the adipogenic differentiation of MSC and the role of BMA in the survival and drug resistance of MM cells. The expression of adipogenic/osteogenic differentiation-related genes PPAR-γ, DLK1, DGAT1, FABP4, FASN and ALP both in MSC and MSC-derived adipocytes was determined with real-time quantitative PCR. The Western blot was employed to detect the expression levels of IL-6, IL-10, SDF-1α, TNF-α and IGF-1 in the supernatant with or without PPAR-γ inhibitor.RESULTS:The results of oil red O staining of bone marrow smears showed that BMA increased significantly in patients of ND-MM compared with the normal control group, and the BMA content was related to the disease status. The content of BMA decreased in the patients with effective chemotherapy. MM cells up-regulated the expression of MSC adipogenic differentiation-related genes PPAR-γ, DLK1, DGAT1, FABP4 and FASN, but the expression of osteogenic differentiation-related gene ALP was significantly down-regulated. This means that the direct consequence of the interaction between MM cells and MSC in the bone marrow microenvironment is to promote the differentiation of MSC into adipocytes at the expense of osteoblasts, and the cytokines detected in supernatant changed. PPAR-γ inhibitor G3335 could partially reverse the release of cytokines by BMA. Those results confirmed that BMA regulated the release of cytokines via PPAR-γ signal, and PPAR-γ inhibitor G3335 could distort PPAR-γ mediated BMA maturation and cytokines release. The increased BMA and related cytokines effectively promoted the proliferation, migration and drug resistance of MM cells.CONCLUSION:The BMA and its associated cytokines are the promoting factors in the survival, proliferation and migration of MM cells. BMA can protect MM cells from drug-induced apoptosis and plays an important role in MM treatment failure and disease progression.

Multiple myeloma (MM) is a lethal B cell neoplasm characterized by clonal expansion of malignant plasma cells in the bone marrow and remains incurable due to disease relapse and drug resistance. Bone marrow adipocytes (BMAs) are emerging as playing active functions that can support myeloma cell growth and survival. The aim of this study is to investigate myeloma-mesenchymal stem cells (MSCs) interaction and the impact of such interactions on the pathogenesis of MM using in vitro co-culture assay. Here we provide evidence that MM cell up-regulated MSCs to express PPAR-γ and pushes MSCs differentiation toward adipocytes at the expense of osteoblasts in co-culture manner. The increased BMAs can effectively enhance MM cell to proliferation, migration, and chemoresistance via cell-cell contact and/or cytokines release regulated by PPAR-γ signal pathway. This effect was partially reversed in medium containing PPAR-γ antagonist G3335 and indicated that G3335 distorts the maturation of MSC-derived adipocytes and cytokines release by adipocytes through inhibition of PPAR-γ, a key transcriptional factor for the activation of adipogenesis, or cell to cell contact, or both. In meantime, we observed higher expression of adipocyte differentiation associated genes DLK1, DGAT1, FABP4, and FASN both in MSCs and MSC derived adipocytes, but the osteoblast differentiation-associated gene ALP was down regulated in MSCs. These finding mean that direct consequence of MM/MSC interaction that play a role in MM pathogenesis. Consistent with those in vitro results, our primary clinical observation also showed that bone marrow samples from MM patients had significantly higher bone adiposity in comparison with controls and the number of adipocytes decreased in those who were response to anti-MM therapy. Our finding suggested that BMAs may have an important contribution to MM progression, particularly in drugs resistant of MM cells, and plays an important contribution in MM bone disease and treatment failure, but more clinical studies are needed to confirm its role.

Diffuse large B-cell lymphomas (DLBCLs) are phenotypically and genetically heterogeneous. Two main subgroups of DLBCL include germinal center B-cell-like (GCB) and activated B-cell-like (ABC). Molecular profiling can further classify DLBCL into four subtypes: MCD (both CD79B and MYD88 L265P), BN2 (NOTCH2 mutation or BCL6 fusion), N1 (NOTCH1 mutation), or EZB (EZH2 mutation or BCL2 fusion). EZH2 inhibitors were recommended for patients with the EZB subtype of DLBCLs; however, little is known about the therapeutic mechanisms. Our results showed that DZNep arrested G1/S phase of GCB-DLBCL cells and inhibited the cell proliferation in vitro through upregulation of p16 by demethylating its promoter. These results suggest that DZNep may have potential as a novel therapeutic agent for DFLBL therapy. This agent may serve as a novel molecular agent to be applied to GCB DLBCL.

Growing evidence shows that cancer progression links with both heterogeneity of the tumor microenvironment and dysregulated activity of immune cells. Cancer-secreted exosomes are being recognized as indispensable mediators of the exchange cargo between cancer and immune cells. The M2-phenotype tumor-associated macrophages have the function of promoting tumor progression and drug resistance. Diffuse large B-cell lymphoma(DLBCL) is a highly heterogeneous and very common malignant non-Hodgkin's lymphoma. Here, we demonstrate that different subtype DLBCL cell-derived exosomes are internalized by macrophages, which can affect macrophages polarization. The mechanism of DLBCL-derived exosomes on macrophage polarization remains unclear currently. This study showed that DLBCL-secreted exosomes could induce the transformation of macrophages to a protumor M2-like phenotype, and block the drug-induced apoptosis of DLBCL cells in an indirect co-culture system. Different DLBCL-derived exosomes could change the phenotype of macrophages through the STAT3 signaling, which upregulated the expression of oncogenic genes and classical markers of M2-like phenotype macrophages, such as IL-10, CD206, and CD163. The addition of DLBCL-derived exosomes resulted in the activation of the STAT3 signaling pathway of M0/M2 macrophages in an indirect co-culture system. GP130 was highly enriched in DLBCL-derived exosomes, which triggered the activation of STAT3 of macrophages and subsequently induced the downstream targets such as BCL2, SURVIVIN, and BAX. The parallel changes of STAT3 and GP130 in macrophages confirmed that GP130 of DLBCL-derived exosomes promoted macrophage polarization by activating STAT3 signaling. Furthermore, all of these effects could be reversed by the GP130 inhibitor SC144. The data indicated that DLBCL-derived exosomes could trigger macrophages polarization into a pro-survival M2-like phenotype, which was at least partially through the GP130/STAT3 signaling pathway. Collectively, this study showed that DLBCL-derived exosomes could promote macrophages transformation to protumor M2-like phenotype in the tumor microenvironment.

INTRODUCTION:Tubulo-interstitial injury is a poor prognostic factor for lupus nephritis (LN). Here, we tested whether iguratimod could inhibit tubulo-interstitial injury in LN. METHODS:MRL/lpr mice, an animal model of lupus, were treated with iguratimod or vehicle solution. Pathological changes of kidney were evaluated blindly by the same pathologist. Renal type I collagen (COL-I), IgG, E-cadherin, fibroblast-specific protein 1 (FSP-1) were detected by immunofluorescence, immunohistochemical staining or quantitative real-time PCR. After treated with transforming growth factor β1 (TGF-β1) and iguratimod, E-cadherin, fibronectin, Smad2/3, p38 MAPK, p-Smad2/3, and p-p38 MAPK, β-catenin and TGF-β type II receptor (TGFβRII) in HK2 cells were measured by western blotting, quantitative real-time PCR or immunofluorescence. RESULTS:Iguratimod reduced immune deposition along the tubular basement membrane, inhibited the tubulo-interstitial infiltration of inflammatory cells, and alleviated tubular injury in MRL/lpr mice. Moreover, Iguratimod eased the tubulo-interstitial deposition of collagen fibers, which was confirmed by decreased expression of COL-I. Furthermore, iguratimod suppressed the expression of FSP-1 and increased that of E-cadherin in renal tubular epithelial cells. In HK2 cells cultured with TGF-β1, iguratimod treatment not only reversed cellular morphological changes, but also prevented E-cadherin downregulation and fibronectin upregulation. In addition, iguratimod inhibited phosphorylation of TGFβRII, Smad2/3 and p38 MAPK in HK2 cells treated with TGF-β1, and also blocked nuclear translocation of β-catenin. CONCLUSION:Iguratimod eased tubulo-interstitial lesions in LN, especially tubulo-interstitial fibrosis, and might have potential as a drug for inhibiting the progression of tubulo-interstitial fibrosis in LN.

目的 观察不同来源间充质干细胞(MSCs)中由连接蛋白43(Cx43)组成的细胞间隙连接(GJIC)及其介导的信号对多发性骨髓瘤(MM)侧群细胞(SP细胞)生物学行为的影响,并探讨其可能机制.方法 分离培养不同来源的间充质干细胞(MSCs);应用流式细胞术分选MM细胞株RPMI 8266的SP细胞;采用RT-PCR技术及蛋白印迹(Western blot)法检测不同来源MSCs、RPMI 8266、SP细胞中Cx43基因及蛋白水平表达;直接共培养观察不同来源MSCs对SP细胞周期、Cx43蛋白表达、体外集落形成能力、干细胞相关基因表达、细胞因子分泌和耐药的变化以及加入连接通道抑制剂18α甘草次酸(α-GA)后的影响.结果 MM-MSCs与ND-MSCs形态及表型无明显区别,与RPMI 8266细胞均表达较高水平的Cx43;与MM-MSCs共培养可使更多SP细胞进入G0期(P<0.001),SP细胞的c-myc、KIF4和SOX2基因表达显著上调,而Oct-4基因表达下调,加入α-GA后,c-myc、KIF4和SOX2均有不同程度下调,但无显著性差别;使Cx43表达上调,分别为(31.00±2)％和(39.00±2)％;使体外集落形成能力上调,加入α-GA可部分抑制该作用;RPMI 8266存在少量c-myc、KIF4、SOX2和Oct-4基因表达,SP细胞亚群中该类基因明显上调,MM-MSCs分泌高水平的白介素(IL)-6,与SP细胞共培养后,其上清液中IL-6、IL-10及TGF-β表达上调(P=0.0072,P=0.037);bFGF和IL-17则无明显变化.加入α-GA后,上清液中IL-6、IL-10和TGF-β水平降低;MM细胞对硼替佐米诱导的凋亡敏感,但SP细胞敏感性较差,与MM-MSCs共培养显著减少硼替佐米介导的细胞凋亡,加入α-GA可部分恢复MM细胞对硼替佐米的敏感性.结论 MM-MSCs与多发性骨髓瘤SP细胞上调Cx43蛋白表达,形成更多GJIC,并通过改变MSCs细胞因子分泌谱,促进SP细胞增殖和耐药,可能是最终导致MM复发的原因之一.

Purpose: Current studies on the mechanism of tumor necrosis factor-like weak inducer of apoptosis (TWEAK) in lupus nephritis (LN) mainly focus on the inflammatory pathway. Herein, we aimed to determine whether TWEAK could promote the progression of renal interstitial fibrosis by regulating peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1 alpha) expression and intervening in lipid metabolism in LN. Materials and Methods: MRL/lpr mice, an animal model of lupus, were treated with the anti-TWEAK antibody or co-treated with adeno-associated virus-mediated PGC-1 alpha short hairpin RNA (shRNA). In addition, human proximal tubular epithelial cells (HK2 cells) were treated with recombinant human TWEAK (rhTWEAK) or ammonium pyrrolidine dithiocarbamate (PDTC) in vitro. Results: The renal contents of free fatty acids and triglycerides were higher in MRL/lpr mice than in MRL/MpJ mice; however, these contents were decreased by treatment with the anti-TWEAK antibody. Based on immunofluorescence staining, the expression of PGC- 1 alpha was markedly more in the renal tubules of MRL/MpJ mice than in the glomeruli. However, treatment with anti-TWEAK antibody increased the levels of PGC-1 alpha and its downstream target genes, which were remarkably lower in MRL/lpr mice than in MRL/MpJ mice. AntiTWEAK antibody effectively eased renal interstitial fibrosis, which manifested as a decrease in the deposition of collagen fibers and the inhibition of type I collagen and fibronectin expression. However, the therapeutic effects of the anti-TWEAK antibody were abolished by PGC-1 alpha shRNA. Treatment with rhTWEAK decreased PGC-1 alpha expression in both dose- and time-dependent manners in HK2 cells in vitro. PDTC, an inhibitor of I kappa B alpha phosphorylation, suppressed the decrease in the PGC-1 alpha protein level induced by rhTWEAK treatment. Conclusion: Our results suggest that TWEAK prevents renal tubular PGC-1 alpha expression by promoting NF-kappa B activation, resulting in a deficiency in lipid metabolism and the progress of renal interstitial fibrosis. The upregulation of renal tubular PGC-1 alpha expression to improve lipid metabolism is one of the mechanisms employed by the anti-TWEAK antibody to treat renal interstitial fibrosis.

OBJECTIVE:To investigate the effect of gap junction intercellular communication (GJIC) combined by connexin43 (Cx43) and its signal to the biobehavior of multiple myeloma (MM) cells, and its possible mechanism. METHODS:The mesenchymal stem cell (MSC) cells were isolated and cultured from patients with MM and normal donors. The expression of connexin43 (Cx43) in MSC cells from different sources was detected by RT-PCR and Western blot. The side population (SP) cells were sorted by flow cytometry (FCM). The effect of MSC cells from different sources to the cell cycle, Cx43 expression, colony formation in vitro, stem cell related genes expression, cytokines secretion and chemoresistance in MM SP cells as well as with or without Cx43 inhibitor 18α-glycyrrhetinic acid (18α-GA) was observed. RESULTS:There was no significantly difference between the MSC isolated from normal donor and MM patients. Western blot showed that Cx43 expression in SP cells was up-regulated when the cells were incubated with MSC, and medium containing 18α-GA could partially inhibit it, moreover, it was more significant in MSC cells of MM patients. The ability of colony formation of SP cells in vitro was higher than those of MM cells and MM-MSC could promote the colony formation in a co-culture manner. The effect of MM-MSC to SP cells was down-regulated after 18α-GA was added. RT-PCR showed that there was several important stem cell-related genes including c-myc, Oct-4 Klf-4, and Sox-2 were found in RPMI 8226 cells, but those cells were up-regulated in SP cells (P<0.001). Meanwhile, MM-MSC could up-regulate the expression of c-myc, Klf-4 and Sox-2 (P<0.001), but down-regulate Oct-4 gene in the SP cells. The expression of those genes decreased after 18α-GA was added, but showed no significant difference (P>0.05). Cytometry bead array assays showed that MM-MSCs could secrete high level of IL-6, but the levels of IL-6, IL-10 and TGF-β increased significantly when the MM-MSCs were co-cultured with SP cells (P<0.05), especially the levels of IL-6 and IL-10 were significantly higher than cultured alone. There was no significant change in the levels of bFGF and IL-17 before and after co-cultured. The levels of IL-6, IL-10 and TGF-β in supernatant decreased significantly after GJ inhibitor 18α-GA was added. PI/Annexin V assay showed that MM cells were sensitive to bortezomib (BTZ)-induced apoptosis, but the sensitivity for SP cells was weaker. The ratio of cell apoptosis was 75.2%±0.77% and 8.12%±0.86% (P<0.001), respectively. MM-MSC could down-regulate the cell apoptosis induced by BTZ, while the sensitivity of MM cells to BTZ could be partially recovered after GJ inhibitor was added. CONCLUSION:MSC derived from MM patients can enhance GJIC to maintain its "hematopoiesis" by up-regulating the expression of Cx43 in MM cells, and at the same time promote cell proliferation and drug recistance by secreting multiple cytokines, which finally contributes to the relapse of MM.

Objective: This study aimed to compare the pharmacokinetic, pharmacodynamic and safety profiles of HLX01（a rituximab biosimilar） and reference rituximab sourced from China（Mab Thera?; rituximab-CN）.Methods: Here we report the results of two phase 1 studies. In the phase 1 a, open-label, dose-escalation study（NCT03218072, CTR20140400）, eligible patients received 250, 375 and 500 mg/m2 HLX01 sequentially at 7-day intervals, after confirming no dose-limiting toxicity（DLT）. In the phase 1 b, double-blind study（NCT02584920,CTR20140764）, eligible patients were given a single dose of 375 mg/m2 HLX01 or rituximab-CN. The primary endpoints included safety and tolerability parameters for the phase 1 a and the area under the plasma concentrationtime curve from time zero to day 91（AUC0-91 d） for the phase 1 b study. Equivalence was concluded if 90%confidence interval（90% CI） for the geometric least squares mean ratio（GLSMR） fell in the pre-specified equivalence criteria（80%-125%）.Results: Between June 20, 2014 and January 5, 2015, 12 patients were enrolled in the phase 1 a study. The pharmacokinetics of HLX01 showed dose proportionality and accumulation to steady state. HLX01 was well tolerated, with no serious adverse events（AEs）, discontinuations or DLTs. Between November 8, 2014 and August13, 2015, 87 eligible patients were enrolled in the phase 1 b study, including 43 who received HLX01 and 44 who were treated with rituximab-CN. The equivalence endpoint was met with GLSMR for AUC0-91 d being 89.6%（90% CI: 80.4%-99.8%）. AEs, anti-drug antibodies, and CD19+ and CD20+ B lymphocyte counts were similar between the HLX01 and rituximab-CN treatment groups.Conclusions: Treatment with HLX01 was safe and well tolerated in Chinese patients with B-cell lymphoma.HLX01 and rituximab-CN have similar pharmacokinetic, pharmacodynamic and safety profiles.

Background: Renal interstitial fibrosis (RIF) is one of the main poor prognostic factors of lupus nephritis (LN). Here, we tested whether iguratimod could inhibit RIF in LN. Methods: MRL/lpr mice, an animal model of lupus, were treated with iguratimod or vehicle solution. Pathological changes of kidney was evaluated blindly by the same pathologist. Renal type I collagen (COL-I), IgG, E-cadherin, fibroblast-specific protein 1 (FSP-1) were detected by immunofluorescence, immunohistochemical staining or quantitative real-time PCR. After treated with transforming growth factor β1 (TGF-β1) and iguratimod, E-cadherin, fibronectin, Smad2/3, p38 MAPK, p-Smad2/3 and p-p38 MAPK in HK2 cells were measured by western blotting, quantitative real-time PCR or immunofluorescence. Results: In MRL/lpr mice, iguratimod eased the renal interstitial deposition of collagen fibers, further confirmed by decreased expression of COL-I after iguratimod treatment. Moreover, upstream pathological processes of RIF, including IgG deposition along the tubular basement membrane, infiltration of inflammatory cells into renal interstitium and tubular injury, were also prevented effectively by iguratimod treatment. Furthermore, iguratimod suppressed the expression of FSP-1 and raised the expression of E-cadherin in renal tubular epithelial cells, suggesting that iguratimod reversed the process of tubular epithelial-to-mesenchymal transition (EMT). In HK2 cells induced by TGF-β1, co-treatment with iguratimod not only reversed cellular morphologic change, but also prevented down-regulation of E-cadherin and up-regulation of fibronectin. Finally, iguratimod blocked TGF-β1-induced phosphorylation of Smad2/3 and p38 MAPK in HK2 cells. Conclusions: Our results suggest that iguratimod shows an inhibitory effect on the progress of RIF in vivo and in vitro, so iguratimod may be a potential drug for the treatment of RIF in LN.

Exosomes are key mediators of intercellular communication and play a role in the pathogenesis and progression of cancer. Exosomes in circulating body fluids serve as molecular markers for cancer diagnosis. This study aimed to investigate the role of exosomal microRNA (miR)-1910-3p in breast cancer and determine its clinical diagnostic value. MiR-1910-3p promoted proliferation and migration of breast cancer cells in vitro and in vivo. In vitro, exosomes enriched in miR-1910-3p transferred miR-1910-3p to mammary epithelial cells and breast cancer cells, promoting proliferation and migration, inhibiting apoptosis, and inducing autophagy. In vivo, exosomes enriched in miR-1910-3p promoted the proliferation and migration of breast cancer cells. MiR-1910-3p downregulated myotubularin-related protein 3, activated the NF-κB and wnt/β-catenin signaling pathway, and promoted breast cancer progression. Serum miR-1910-3p in exosomes was an effective diagnostic marker that improved the sensitivity of breast cancer diagnosis when used in combination with the traditional tumor marker CA153. In conclusion, breast cancer cell-derived exosomes promoted the growth, metastasis, and autophagy of breast cancer cells by transferring miR-1910-3p. MiR-1910-3p in serum exosomes may serve as a novel molecular marker for breast cancer diagnosis.

Background/Aims: This study aimed to investigate the level of soluble tumor necrosis factor-like weak inducer of apoptosis (sTWEAK) and its correlation with micro-inflammation and atherosclerosis in continuous ambulatory peritoneal dialysis (PD) patients. Methods: This retrospective study involved 23 healthy subjects (control group), 23 hemodialysis (HD) patients (HD group) and 26 PD patients (PD group). Serum biochemical measurements and sTWEAK assessments were tested. The association between intima-media thickness (IMT) and sTWEAK concentrations was evaluated. Results: The TWEAK level was lower in PD (155.16 ± 3.69 pg/mL, p < 0.001) and the HD group (150.16 ± 7.23 pg/mL, p < 0.001) than that in the control group (193.05 ± 5.36 pg/mL), with no significant difference between the PD group and the HD group. In the PD and HD groups, sTWEAK was significant negatively correlated with CPR, fibrinogen, and white blood cell (p < 0.05). Besides, compared to lower sTWEAK concentration end-stage renal disease (ESRD) patients (no >161.9 pg/mL), patients who had a higher level of sTWEAK (>161.9 pg/mL) had a lower IMT (0.97 ± 0.04 vs. 0.84 ± 0.03 cm, p = 0.029). After adjusted for sex, age, hypertension, diabetes, duration of dialysis, triglyceride, total cholesterol, low-density lipoprotein, and serum glucose, sTWEAK (B = –0.002, r = 0.015) and CRP (B = 0.022, r = 0.015) were independent risk factors for the IMT of ESRD patients. Conclusion: Plasma TWEAK is inversely associated with carotid IMT among patients undergoing HD and PD.

AIM In clinical practice, an anechoic signal was often exhibited between the volar plate (VP) of the proximal interphalan-geal joint (PIPJ) (PIPJVP) and the flexor digitorum tendon (FDT) on ultrasound, which suggests the presence of effusions (PIPJVP-FDT effusions). The purpose of this study was to investigate the prevalence of PIPJVP-FDT effusions and to explore the possible mechanism preliminarily. MATERIAL AND METHODS A single-center, cross sectional study in hand osteoarthritis (HOA) patients, rheumatoid arthritis (RA) patients and healthy controls was conducted. Ultrasound examination was per-formed by the same real-time scanner with 18-MHz linear array transducer. Bilateral interphalangeal joints (IPJs) of the thumb, 2ed, 3rd, 4th and 5th PIPJs were examined. The PIPJVP-FDT effusions was defined as an anechoic signal between the PIPJVP and FDT in two perpendicular ultrasound planes. RESULTS In total, 200 patients with HOA, 78 patients with RA and 101 healthy controls were eligible for the study. 37.6% of healthy controls and 35.0% of HOA patients showed PIPJVP-FDT effusions, while only 11.5% of RA patients had PIPJVP-FDT effusions (p<0.001). The 2ed, 3rdand 4th PIPJs showed more PIPJVP-FDT effusions, while the IPJs of the thumbs and 5th PIPJs showed less PIPJVP-FDT effusions (p<0.05). Furthermore, the prevalence of PIPJVP-FDT effusions in different age groups were similar in HOA patients and healthy controls. CONCLUSION To the best of our knowledge, this paper is the first to demonstrate that the presence of PIPJVP-FDT effusions is a very common phenomenon in HOA patients and healthy individuals, and may be unrelated to inflammation, degeneration and age.

Introduction The pro-tumoural role of M2 macrophages has been described in various cancers and is associated with poor clinical outcomes. Exosomes derived from cancers can shuttle information among tumour cells, immune cells, and the microenvironment, leading to microenvironment remodelling. The mechanism involved in exosomes influencing macrophages polarisation is still unclear. Material and methods We established a macrophage model derived from THP-1 monocytes. To mimic the in vivo microenvironment, we generated an indirect co-culture system of macrophages with diffuse large B-cell lymphoma (DLBCL) cell lines, OCI-LY1 and OCI-LY3 cells. In addition, pre-treated macrophages with exosomes derived from OCI-LY1/3 lymphoma cells were used in our study. The changes in macrophage polarisation stage, surface receptor expression, and cytokine expression at the mRNA level were analysed. Moreover, the effect of macrophages on drug-induced lymphoma cell apoptosis in the indirect co-culture system was assessed. Furthermore, protein analysis of NF-κB and p-p65 in macrophages exposed to exosomes was performed by western blot. Results DLBCL-derived exosomes had different effects on macrophages at different differentiation stages; these macrophages had different effects on drug-induced apoptosis of lymphoma cells in indirect co-culture. Moreover, NF-κB signalling molecules were involved in this process. Conclusions The results indicate that DLBCL-derived exosomes can lead to more M2 phenotypes, promote tumour growth, and play a substantial role in tumour progression, invasion, and drug-resistance.

目的:采用条件性基因敲除技术构建造血系统间隙连接蛋白43(Cx43)基因敲除(Cx43-/-)小鼠模型,并探讨Cx43在维持造血细胞自我更新及功能稳定中的作用.方法:将引进的2对转基因小鼠Cx43 loxP/loxP和Lyz-Cre/+杂交,选取F1雌性子代Cx43 loxP/-_Lyz-Cre/+与雄性Cx43 loxP/loxP合笼回配,提取所获得子代小鼠鼠尾组织基因组DNA,采用PCR方法鉴定小鼠基因型,RT-PCR方法筛选Cx43-/-小鼠,同时分析小鼠不同器官中Cx43基因的表达差异;该类小鼠经5-氟尿嘧啶(5-FU;125 mg/kg)处理,在化疗前及化疗后第5、10和15天经眼球取血分析其血象变化.Cx43-/-及Cx43+/+小鼠予7.5 Gy(60 Co-γ)的致死量照射,剂量率1 Gy/min,照射后6 h分别给予事先准备就序的骨髓细胞,每只3×106细胞于尾静脉注入,2周后处死小鼠检测造血是否重建:分离股骨切片后,收集骨髓细胞进行细胞表型分析(选用的单抗为CD45R、Gr-1、CD4、CD8a、TCRαβ、Mac-1、抗sIgM、TER119、Sca-1及CD117);同时进行体外造血细胞集落实验观察造血细胞的体外增殖能力.结果:本研究通过2种转基因小鼠间杂交和回交,成功获得造血系统选择性Cx43基因敲除小鼠;该类小鼠骨髓及外周血细胞无Cx43表达,参与造血的组织,如肝脏和脾脏中Cx43表达也显著下调(P<0.01),而心脏和肾脏的Cx43表达则无影响,小鼠成年后外周血象分析并无明显异常,但应急代偿能力下降,经5-FU处理后,其造血功能恢复显著减缓,处理15 d后,Cx43+/+小鼠造血功能已接近正常水平,而Cx43-/-小鼠仍无明显的恢复迹象,血红蛋白、白细胞及血小板仍处低位,2者差别有统计学显著性(P<0.01);体外集落试验也证实Cx43-/-小鼠造血干/祖细胞的增殖能力下降,其CFU-GM或CFU-E集落数均明显少于Cx43 Lin-/c-Kit+/Sca-1+/+小鼠(P<0.01),但流式细胞术结果显示,Cx43-/-小鼠骨髓中+细胞亚群数量与Cx43+/+小鼠相比差异并无统计学显著性;Cx43-/-小鼠在化疗或移植后其骨髓造血功能重建均延迟,且化疗15 d后骨髓切片及涂片均证实其骨髓中造血细胞增生程度明显降低,脂肪组织显著增多,而且T、B细胞发育也有异常.此外,其外周血中CD4 CD4+CD8+细胞比例比野生型小鼠增多(P<0.05),但+T细胞显著减少(P<0.01),尤其是TCRαβ亚群细胞减少最为明显(P<0.01).同样,Cx43 CD45R+sIgM-/-小鼠外周血中-细胞亚群比例与野生型小鼠相比显著减少(P<0.01).结论:骨髓中Cx43基因表达在造血干/祖细胞发育(尤其是应急状态时)具重要作用,敲除Cx43基因后造血干/祖细胞增殖减缓,造血及免疫重建功能受损.

Background Mesenchymal stem cells (MSCs) represent a subset of non-hematopoietic adult stem cells, which can also fuse with other cells spontaneously in bone marrow and capable of adopting the phenotype of other cells. The fusion of somatic cells with stem cells can reprogram somatic cells to a pluripotent state. Our research on the fusion of bone marrow mesenchymal stem cells(BM-MSCs) and MM cells demonstrate that the fused cells can exhibit stemness and cancer cell-like characteristics. Results We successfully produced a hybrid cells that acquired larger size and multinucleation, in which partial chromatin condensation, a visible nucleolus, and one or more round or oval nucleus. Experiments results showed that the stemness markers highly expressed in these fused cells and there were much more chromosomes in fused cells than those in parental cells as well as exhibited increased resistance to drug treatment. Conclusions Our results suggest that cell fusion between BM-MSCs and MM cells could contribute it genomic heterogeneity and play a role on disease progression. Methods We fused human BM-MSCs with MM cells lines RPMI 8226 or XG1 in vitro by polyethylene glycol (PEG), and the hybrid cells were sorted by sedimentation assays. The growth, migration, cell cycle, chromosome and drug sensitive of hybrids were assessed by cell counting, cell colony formation, transwell assays, cytogenetic assay and flow cytometry (FCM). The proteins and genes related to stemness and cytokines were tested by western blot and/or real-time quantitative RT-PCR.

To assess the efficacy of long-term treatment with nonsteroidal antiinflammatory drugs (NSAIDs) on bone marrow oedema (BMO) of the sacroiliac joint in newly diagnosed axial spondyloarthritis (SpA) with a symptom duration of less than 4 years, a single-center, open-label study in a cohort of consecutive patients with newly diagnosed axial SpA was conducted. Eligible patients had magnetic resonance imaging (MRI)-determined BMO of the sacroiliac joint at baseline, had a symptom duration of less than 4 years, and were naïve to NSAIDs. After the baseline MRI, an optimal dose of NSAID was administered for 24 or 48 weeks. BMO of sacroiliac joint was quantified by applying the Spondyloarthritis Research Consortium of Canada (SPARCC) system. Disease activity was expressed using the Ankylosing Spondylitis Disease Activity Score (ASDAS). Primary end points were improvement in BMO of sacroiliac joint at week 24 or week 48. Forty-three patients were recruited, and 33 patients eventually completed the study, including 10 patients having follow-up MRI at week 24 and 23 patients having follow-up MRI at week 48. Overall, the mean of SPARCC score decreased from 21.8 ± 16.1 at baseline to 10.2 ± 12.8 at follow-up (p < 0.001). 75.8% of the patients displayed a minimally important change, and 30.3% became free of BMO. The mean of ASDAS-CRP decreased from 3.1 ± 1.0 at baseline to 2.1 ± 1.0 at follow-up (p < 0.001). Long-term treatment with optimal dose NSAIDs could significantly alleviate BMO of sacroiliac joint in early and active axial SpA.

OBJECTIVES:To investigate the dynamic change of follicular T helper cells (TFH) in patients with malignant lymphoid disease (MLD) and to explore its clinical significance.METHODS:The dynamic change of TFH cells, ICOS+- and PD-1+ TFH cells at pretreatment and different treatment periods was determined by flow cytometry in 85 MLD patients. Concentration of interleukin 21 (IL-21) was evaluated by ELISA, and the correlation between clinical prognosis and the ratio of TFH cells was analyzed.RESULTS:Significantly increased ICOS+- and PD-1+ TFH cells were found in MLD patients at pretreatment compared to healthy controls. Decreased or even close to normal levels of ICOS+- and PD-1+ TFH cells were found at the end of treatment. However, in the patients with progressive disease, high levels of ICOS+- and PD-1+ TFH cells were found. Moreover, a significantly increased plasma IL-21 level was found in MLD patients. Negative correlation was found between the level of ICOS+-, PD-1+ TFH cells, as well as IL-21 and the prognosis of MLD.CONCLUSIONS:Significantly increased TFH cell ratios were found in patients with MLD, and decreased TFH cells ratios could be expected in those treatment-effective patients, which could be used as the therapeutic efficacy index.

Objective To explore the breeding,genotyping and mechanism of the onset of myelodysplasia in the conditional connexin 43(Cx43) knock out mice(Cx43KO mice).Methods Two pairs of transgenic mice(Cx43loxP/loxP and Lyz-Cre/+) were inbreeded to produce Cx43loxP/-_Lyz-Cre/+ mice,females of which were used to mate with the male Cx43loxP/loxP mice to finally get the Cx43loxP/loxP_Lyz-Cre/+ mice(Cx43KO mice).After genome DNA was extracted from the mice tails,the genotype was identified by PCR and agarose gel electrophoresis.The real-time PCR was used to detect the accuracy of the Cx43KO mice.Results Both breeding and reproduction of Cx43KO mice were successful.It was fruitful to obtain four genotype mice of Cx43loxP/loxP_Lyz-Cre/+,Cx43loxP/-_Lyz-Cre/+,Cx43loxP/loxP and Cx43loxP/-.The results of breeding were met with the Mendel's law.The expressions of Cx43 mRNA in bone marrow,liver and spleen were lower in Cx43KO mice than those in heterozygote ones(Cx43loxP/-_Lyz-Cre/+)(P<0.05).ConclusionThe PCR method is able to correctly identify the genotype of the offspring.Mating female Cx43loxP/-_Lyz-Cre/+ mice with male Cx43loxP/loxP ones is a useful approach to obtain the Cx43KO mice.