Knee osteoarthritis(KOA)is a common chronic degenerative bone and joint disease characterized by degeneration and wear of knee cartilage.It is commonly found in middle-aged and elderly people and seriously impacts on their lower limb activity and quality of life.At present,the treatment of early and middle stage KOA mainly relies on the conservative methods such as oral medication,joint injection,topical patches and traditional Chinese medicine.Platelet rich plasma(PRP),as an autologous platelet concentrate,is rich in various growth factors and has no risk of immune rejection.In recent years,it has been widely used in the repair of bone,joint,and soft tissue injuries.However,the short biological half-life of growth factors in PRP and the fluidity of injection sites can result in insufficient binding force,short action time,poor target therapy efficacy,and the need for repeated injections in the joint cavity,which will increase the risk of iatrogenic infections.Hydrogels are cross-linked polymer networks containing water,and their high histocompatibility and drug release have attracted much attention.The slow and continuous release of drug is achieved by loading PRP onto hydrogel.Its unique adhesion reduces the flow of drug in the joint,thus extending the local action time of PRP and reducing the need for repeated injection.This article reviews the biological characteristics of PRP and hydrogel,the mechanism of action and clinical application of PRP loaded hydrogel in the treatment of KOA,and analyzes the existing problems and challenges,aiming to provide more effective treatment options for KOA patients through the in-depth discussion of this new treatment method.

[摘 要] 目的：探讨四氧化三铁（Fe3O4）纳米粒子（PION）作为药物载体增强二氢卟吩e6（chlorin e6，Ce6）在胶质瘤中的增效作用。方法：采用高温降解法和相转移法制备PEG-Fe3O4@Ce6复合纳米粒子（PION@E6），用水合粒径分析、透射电镜、胶体稳定性分析、紫外可见光吸收光谱等方法对PION@E6进行鉴定。CCK-8法检测胶质瘤U251细胞的增殖活性，流式细胞术检测细胞的凋亡水平，DCFH-DA探针法检测细胞中活性氧（reactive oxygen species，ROS）的水平。构建BALB/c-nu裸鼠胶质瘤U251细胞移植瘤模型，动物活体荧光成像术及磁共振成像（MRI）观察PION@E6及Ce6在移植瘤中的潴留时间，比较PION@E6声动力治疗组及Ce6声动力治疗组的第28天生存情况及肿瘤体积。结果：PION@E6的核心粒径为10 nm、水合粒径为（37.86±12.90）nm，具有良好的水溶性和稳定性；吸收光谱及XRD图谱显示Ce6已经负载到Fe3O4纳米粒子上。与Ce6声动力组比较，PION@E6声动力组U251细胞的增殖活性显著下降（P<0.05），细胞凋亡率显著升高（均P<0.05），细胞中ROS水平显著升高（P<0.05）。荷瘤裸鼠胶质瘤U251细胞移植瘤治疗实验结果显示，与Ce6声动力治疗组比较，PION@E6声动力治疗组裸鼠移植瘤组织中潴留时间显著延长（P<0.05），存活的裸鼠数显著增多，移植瘤体积显著缩小（P<0.01）。结论：Fe3O4纳米粒子对Ce6介导的胶质瘤U251细胞声动力治疗具有明显的增效作用。

[摘 要] 目的：探讨超微氧化铁纳米粒子（ultrasmall iron oxide nanoparticle，USIONP）对人肝细胞癌HepG2细胞迁移和侵袭的影响及其可能的机制。方法：采用粒径分析仪和透射电镜分别分析USIONP的水合粒径和核心粒径，Zeta电位和胶体稳定性实验分析USIONP的分散性及其稳定性以鉴定USIONP的成功制备；用不同质量浓度USIONP（0、50、100、200 μg/ml）或200 μg/ml USIONP+5 mmol/L 3-MA（自噬抑制剂）联合处理HepG2细胞，CCK-8法检测HepG2细胞的增殖活力，Transwell法检测细胞的迁移和侵袭能力，WB实验检测自噬标志物Beclin1、LC3、p62的表达，2’,7’-二氯二氢荧光素二醋酸（DCFH-DA）法测定细胞内活性氧（ROS）水平，铁离子比色法检测细胞内铁离子水平。结果：USIONP的平均水合粒径为（37.86±12.90） nm、核心粒径约10 nm，Zeta电位为–23.8 mV，有良好的水溶分散性，证实了USIONP的成功制备。随USIONP质量浓度升高和处理时间延长，HepG2细胞的增殖活力明显降低（均P<0.05）；与对照组相比，200 μg/ml USIONP处理HepG2细胞24 h后，迁移、侵袭细胞数量均显著减少（均P<0.05），而3-MA能够部分抵消上述影响（均P<0.05）。与对照组相比，100、200 μg/ml USIONP处理组的HepG2细胞中Beclin1和LC3Ⅱ蛋白相对表达水平均显著升高（均P<0.05），而p62蛋白表达水平下降（均P<0.05）；200 μg/ml USIONP可显著提高细胞内ROS水平与铁离子水平，而加入3-MA可阻断其作用（均P<0.05）。结论：USIONP能促进HepG2细胞发生自噬，而自噬通路激活后降解USIONP释放铁离子和导致细胞ROS水平升高，从而抑制HepG2细胞的迁移和侵袭。

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 investigate the impact of down-regulation of histone methyltransferase enhancer of zeste homolog 2 (EZH2) on RUNX3 expressions,proliferation and apoptosis in human pancreatic cancer.Methods The expressions of EZH2 and RUNX3 in 38 pancreatic cancer patients and human pancreatic cancer AsPC-1,PANC-1 and BxPC-3 cells were detected by immunohistochemistry and western blot,respectively.Cells were transfected with siEZH2 by lipofectamin 2000.Real time-PCR and western blot were used to detect EZH2 and RUNX3 expressions.Cell growth and apoptosis in vitro and vivo were assessed by MTT,flow cytometry and nude mice experiments,respectively.The correlation among the expressions of EZH2,clinical pathological features and overall survival rate were analyzed.Results Elevated EZH2 and decreased RUNX3 expressions were observed in human pancreatic cancer tissues and cells (P ＜ 0.05).Knockdown of EZH2 reduced cell growth and induced apoptosis in vitro and vivo by upregulating RUNX3 protein expression (P ＜ 0.05).In addition,the EZH2 expressions were correlated with tumor stage,lymph node metastasis and poor prognosis (P ＜ 0.05).Conclusions EZH2 expressions were correlated with malignancy and poor prognosis in pancreatic cancer.Tumor cell proliferation was promoted by EZH2 through down-regulation of RUNX3.EZH2 may be a potential therapeutic target for pancreatic cancer.

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.

Objective To evaluate the reliability of autologous blood withdrawal during cesarean section. Methods Fifteen patients preoperatively diagnosed with pernicious placenta previa and∕or accrete by using ultrasound and magnetic resonance imaging, aged 20-35 yr, weighing 55-75 kg, at≥36 weeks of gestation, were enrolled in the study. Blood containing amniotic fluid from the surgical field was collected, and the washed blood was processed using cell?salvage machine and then filtered using a leukocyte depletion filter during cesarean section. The 20 ml blood samples collected included maternal central venous blood after delivery of fetus, unwashed blood, washed blood and filtered blood. The fetal squamous cells were counted using papanicolaou staining. The concentrations of a?fetoprotein, tissue factor, endothelin?1 and histamine were measured by enzyme linked immunosorbent assay. The fetal red blood cells were counted using the acid elution method and HE staining. Results Compared with unwashed samples, the tissue factor concentrations were significantly increased, and the fetal squamous cell count, concentrations of a?fetoprotein and endothelial?1, and fetal red blood cells were decreased in the washed samples. Compared with washed samples, the fetal squamous cell count, concentrations of a?fetoprotein and fetal red blood cells were significantly decreased in filtered samples. Compared with maternal venous blood samples, the tissue factor concentrations were significantly increased, and the fetal squamous cell count and concentrations of a?fetoprotein and endothelial?1 were decreased in filtered samples. Conclusion Autologous blood withdrawn during cesarean section can be used for reinfusion in cesarean section.

BACKGROUND:Currently, the research about effect of non-dextran coated superparamagnetic iron oxide nanoparticles on cellproliferation and cytotoxicity is relatively much less. OBJECTIVE:To evaluate the effects of 0, 25, 50, 75, 100 mg/L non-dextran coated superparamagnetic iron oxide nanoparticles on the proliferation and cytotoxicity of rat bone marrow mesenchymal stem cels. METHODS:Culture media containing 0, 25, 50, 75, 100 mg/L non-dextran coated superparamagnetic iron oxide nanoparticles were prepared for culture of bone marrow mesenchymal stem cels. After 24 hours of culture, the cels were confirmed using Prussian blue staining, and cellcounting was detected using cellcounting kit-8. Meanwhile, lactate dehydrogenase activity in the supernatant and intracelular superoxide dismutase activity were detected. RESULTS AND CONCLUSION:Loading of non-dextran coated superparamagnetic iron oxide nanoparticles in BMSCs was confirmed by Prussian blue staining. The percentage of cels labeled with non-dextran coated superparamagnetic iron oxide nanoparticles was up to 100% when the cels were incubated with a non-dextran coated superparamagnetic iron oxide nanoparticle solution of 50 mg/L and above, but 25 mg/L was insufficient to label al of the cels. Furthermore, as the concentration of non-dextran coated superparamagnetic iron oxide nanoparticles decreased, the cellproliferation rate decreased gradualy. The 25 mg/L group had a minimum cellproliferation rate, but the 25 and 50 mg/L groups showed no statisticaly significant difference (P > 0.05). Therefore, 50 mg/L is considered as the appropriate concentration of non-dextran coated superparamagnetic iron oxide nanoparticles, under which, the labeling efficiency is higher and the cytotoxicity is lower.

Objective To explore the constituent ratio and clinical characteristics of multifocal thyroid papillary carcinoma (MPTC).Methods The clinical data of 1616 cases of papillary thyroid carcinoma(PTC) were retrospectively analyzed from January 2002 to December 2011 of the First Affiliate Hospital of Dalian Medical University,which operated at the first time and confirmed by pathology.The change of constituent ratio of MPTC in PTC was analyzed and the differences of the clinical characteristics of the multifocal group and single focal group were analyzed.Results The constituent ratio of MPTC in PTC was increasing from 8.33％ (4/48) in 2002 to 30.38％ (96/316) in 2011 gradually.Compared to the single focal group,MPTC group had higher rate of neck lymph node metastasis(45.09％ vs 25.02％ ; P =0.000) and extrathyroidal invasion (20.95％ vs 9.04％ ;P =0.000).Compared with pure microcarcinoma,none pure microcarcinoma had higher rate of neck lymph node metastasis (P =0.000).More than two focuses has higher rate of neck lymph node metastasis than two focuses (P =0.000).The rate of recurrence with lymph node metastasis was higher than that without lymph node metastasis (24.05％ vs 8.98％ ;P =0.000) conformed by postoperative pathology.Conclusions The number and constituent ratio of MPTC in PTC is increasing gradually.MPTC group has high proportion of lymph node metastasis and extrathyroidal invasion than single focal group.None pure microcarcinoma has higher rate of neck lymph node metastasis than pure microcarcinoma;more than two focuses has higher rate of neck lymph node metastasis than two focuses.The recurrence rate is correlated with the rate of lymph node metastasis.