Degradation of cartilage by macrophages in culture: Evidence for the involvement of an enzyme which is associated with the cell surface

A cell culture system is described in which purified mononuclear phagocytes may be cultured with a cartilage substrate which is radiolabelled in its proteoglycan. Resident mouse peritoneal macrophages degraded this substrate, and did so more avidly if cultured in direct contact with it. There was no evidence for complete intralysosomal degradation of the proteoglycan of the cartilage. Lysates were found to contain considerable activity at pH 7, which was inhibited by the presence of 10% serum, or by boiling the lysate.

Proximity of macrophages to the substrate did not induce selective release of the lysosomal marker enzyme hexosaminidase, and concentrated enzymes secreted from the macrophages after treatment with the lysosomotropic agent ammonium chloride were ineffective in degrading cartilage at neutral pH.

The active enzyme in macrophage lysates at neutral pH was found to be sedimentable by 100,000 × g centrifugation for 1 hour, in absence of lysosomal protective agents. There is evidence for a cell membrane-associated process in the degradation of cartilage by these cells, which may be a proteolytic, endoglycosidic or free radical-mediated event.     

Chitosan/gelatin porous scaffolds containing hyaluronic acid and heparan sulfate for neural tissue engineering

The novel chitosan (Cs)/gelatin (Gel) porous scaffolds containing hyaluronic acid (HA) and heparan sulfate (HS) were fabricated via freeze-drying technique, and their physicochemical characteristics including pore size, porosity, water absorption, and in vitro degradation and biocompatibility were investigated. It was demonstrated that the Cs/Gel/HA/HS composite scaffolds had highly homogeneous and interconnected pores with porosity above 96% and average pore size ranging from 90 to 140?μm and a controllable degradation rate. The scanning electron microscopic images, cell viability assay, and fluorescence microscopy observation revealed that the presence of HA and HS in the scaffolds significantly promoted initial neural stem and progenitor cells (NS/PCs) adhesion and supported long-time growth in three-dimensional environment. Moreover, NS/PCs also maintained mutilineage differentiation potentials with enhanced neuronal differentiation upon induction in the Cs/Gel/HA/HS composite scaffolds in relation to Cs/Gel scaffolds. These results indicated that the Cs/Gel/HA/HS composite scaffolds were suitable for neural cells’ adhesion, survival, and growth and could offer new and important options for neural tissue engineering applications.     

赛肤润与硫酸镁在小儿甘露醇外渗治疗中的对比研究

目的:比较赛肤润与硫酸镁用于小儿甘露醇外渗的治疗效果,为临床治疗小儿甘露醇外渗提供参考。方法:将60例外周静脉输注甘露醇时发生外渗的患儿随机分为观察组和对照组各30例,出现外渗后两组患儿均立即停止输液,原针头连接无菌注射器抽吸外渗药液。对照组采用硫酸镁治疗,每天3次,每次30 min;观察组采用赛肤润治疗,根据部位大小每次喷1滴或2滴赛肤润,涂抹均匀并按摩3~5 min,每隔4 h 1次,两组观察治疗时间均为72 h,72 h后无效者更改治疗方案。结果:观察组治疗总有效率为100.00%,对照组治疗总有效率73.33%,差异有统计学意义(P<0.05)。结论:赛肤润用于小儿甘露醇外渗的治疗效果优于硫酸镁,且操作简单、安全。     

Development of calcium phosphate/calcium sulfate biphasic biomedical material with hyaluronic acid containing collagenase and simvastatin for vital pulp therapy

ObjectiveIn vital pulp therapy (VPT), a barrier is created with appropriate capping to protect the remaining pulp and thus maintain pulp vitality. Here, we evaluated the feasibility of a biphasic calcium phosphate cement (CPC)–calcium sulfate hemihydrate (CSH) biomaterial containing simvastatin (Sim) and collagenase (Col) for VPT.MethodsCombinations of varying CPC and CSH concentrations were analyzed for their handling properties and setting times, with their structures observed through scanning electron microscopy–energy dispersive X-ray spectrometry (SEM-EDS). Drug release patterns of simvastatin and collagenase combined with CPC–CSH (CPC–CSH–Sim–Col) were also analyzed, followed by biocompatibility and bioactivity tests on human dental pulp stem cells (hDPSCs) and in vivo animal study in canine models; the in vivo results were obtained through microcomputed tomography and histological analysis.ResultsThe results revealed that 70 wt% CPC (CPC7) with 30 wt% CSH (CSH3) exhibited optimal setting time and porous structure for clinical use. The cell viability and cytotoxicity analysis demonstrated that CPC7–CSH3 with or without simvastatin or collagenase did not injure hDPSCs. In vivo, the CPC7–CSH3–Sim–Col induced dentin bridge formation.SignificanceCPC7–CSH3–Sim–Col in this study has great potential as a VPT biomaterial to enhance the dentin bridge formation.     

Comparison of pregnenolone sulfate,pregnanolone and estradiol levels between patients with menstrually-related migraine and controls: an exploratory study

BackgroundNeurosteroids affect the balance between neuroexcitation and neuroinhibition but have been little studied in migraine. We compared the serum levels of pregnenolone sulfate, pregnanolone and estradiol in women with menstrually-related migraine and controls and analysed if a correlation existed between the levels of the three hormones and history of migraine and age.MethodsThirty women (mean age ± SD: 33.5 ± 7.1) with menstrually-related migraine (MM group) and 30 aged- matched controls (mean age ± SD: 30.9 ± 7.9) participated in the exploratory study. Pregnenolone sulfate and pregnanolone serum levels were analysed by liquid chromatography-tandem mass spectrometry, while estradiol levels by enzyme-linked immunosorbent assay.ResultsSerum levels of pregnenolone sulfate and pregnanolone were significantly lower in the MM group than in controls (pregnenolone sulfate: P = 0.0328; pregnanolone: P = 0.0271, Student’s t-test), while estradiol levels were similar. In MM group, pregnenolone sulfate serum levels were negatively correlated with history of migraine (R² = 0.1369; P = 0.0482) and age (R² = 0.2826, P = 0.0025) while pregnenolone sulfate levels were not age-related in the control group (R² = 0.04436, P = 0.4337, linear regression analysis).ConclusionLow levels of both pregnanolone, a positive allosteric modulator of the GABAA receptor, and pregnenolone sulfate, a positive allosteric modulator of the NMDA receptor, involved in memory and learning, could contribute either to headache pain or the cognitive dysfunctions reported in migraine patients. Overall, our results agree with the hypothesis that migraine is a disorder associated with a loss of neurohormonal integrity, thus supporting the therapeutic potential of restoring low neurosteroid levels in migraine treatment.Supplementary InformationThe online version contains supplementary material available at 10.1186/s10194-021-01231-9.     

硫酸沙丁胺醇吸入气雾剂微生物限度检查方法的建立和验证

目的建立和验证硫酸沙丁胺醇吸入气雾剂的微生物限度检查方法。方法根据2015年版《中国药典(四部)》非无菌产品微生物限度检查方法,采用薄膜过滤法测定试验菌株的回收率,确立需氧菌总数、霉菌和酵母菌总数计数方法、控制菌检查方法,并进行验证。结果需氧菌、霉菌和酵母菌总数计数采用薄膜过滤法时,回收率均在0.5~2.0范围内;控制菌的检查采用常规法,结果均符合2015年版《中国药典(四部)》的规定。结论该方法准确、可靠,可用于硫酸沙丁胺醇吸入气雾剂的微生物限度检查。     

Immunostimulatory DNA ameliorates experimental and spontaneous murine colitis

BACKGROUND & AIMS: Impaired mucosal barrier, cytokine imbalance, and dysregulated CD4(+) T cells play important roles in the pathogenesis of experimental colitis and human inflammatory bowel disease. Immunostimulatory DNA sequences (ISS-DNA) and their synthetic oligonucleotide analogs (ISS-ODNs) are derived from bacterial DNA, are potent activators of innate immunity at systemic and mucosal sites, and can rescue cells from death inflicted by different agents. We hypothesized that these combined effects of ISS-DNA could inhibit the damage to the colonic mucosa in chemically induced colitis and thereby limit subsequent intestinal inflammation. METHODS: The protective and the anti-inflammatory effect of ISS-ODN administration were assessed in dextran sodium sulfate-induced colitis and in 2 models of hapten-induced colitis in Balb/c mice. Similarly, these effects of ISS-ODN were assessed in spontaneous colitis occurring in IL-10 knockout mice. RESULTS: In all models of experimental and spontaneous colitis examined, ISS-ODN administration ameliorated clinical, biochemical, and histologic scores of colonic inflammation. ISS-ODN administration inhibited the induction of colonic proinflammatory cytokines and chemokines and suppressed the induction of colonic matrix metalloproteinases in both dextran sodium sulfate- and hapten-induced colitis. CONCLUSIONS: As the colon is continuously exposed to bacterial DNA, these findings suggest a physiologic, anti-inflammatory role for immunostimulatory DNA in the GI tract. Immunostimulatory DNA deserves further evaluation for the treatment of human inflammatory bowel disease.     

纳米羟基磷灰石/硫酸钙复合人工骨顺铂缓释系统注射液的制备与研究

目的研制可注射α-CSH-nano-HA/PHBV-PEG顺铂释药系统,为骨转移瘤提供新型的局部药物缓释系统。方法α-CSH-nano-HA/PHBV-PEG载顺铂制成可注射用α-CSH-nano-HA/PHBV-PEG cis-platinum缓释微球,研究其结构、释药特性、可注射性以及力学性能。结果(1)第1、3、5、7天缓释微球的释药浓度分别为97.5、90.7、83.2、68.5μg/mL,第7天后趋于稳定。(2)可注射α-CSH-nano-HA/PHBV-PEG顺铂释药系统在液固比为0.7时可注射性强,与此同时缓释药系统随着液固比的增大凝固时间延长。结论α-CSH-nano-HA/PHBVPEG顺铂释药系统具有良好的注射性能和缓释作用。     

选择性β肾上腺素受体激动剂对人滑膜细胞增殖能力的影响

目的：观察β-肾上腺素受体( AR)激动药对人成纤维样滑膜细胞( FLS)增殖的作用,探讨β-AR信号对人FLS功能的影响。方法组织块培养法培养人FLS;使用肿瘤坏死因子α( TNF-α)作为刺激剂,MTT法分别检测β1-AR选择性激动剂多巴酚丁胺和β2-AR选择性激动剂沙丁胺醇对人FLS增殖作用的影响,并计算增殖抑制率;使用沙丁胺醇+β2-AR选择性拮抗剂ICI 118551观察其对FLS增殖功能的影响;Western blot法检测人FLSβ-AR、G蛋白偶联受体激酶2(GRK2)和β抑制蛋白2(β-arrestin2)胞膜蛋白的表达水平。结果 TNF-α可以显著促进人FLS的增殖;β2-AR激动剂沙丁胺醇能明显抑制FLS的增殖,其拮抗剂ICI 118551能显著拮抗沙丁胺醇的作用;β1-AR选择性激动剂多巴酚丁胺对FLS的增殖无显著影响;TNF-α和沙丁胺醇均能显著降低β2-AR胞膜表达,上调 GRK2、β-arrestin2的胞膜表达。结论β2-AR信号的激活可以抑制FLS增殖;TNF-α和沙丁胺醇可以使GRK2、β-arrestin2的胞膜表达增加,β2-AR胸膜表达下降。     

Age-related changes and sex differences in chondroitin sulfate isomers and hyaluronic acid in normal synovial fluid

The effects of factors such as age and sex on the metabolism of chondroitin sulfate (CS) and hyaluronic acid (HA) in knee joint tissues are believed to be profoundly important in the onset of joint diseases including osteoarthritis. To test whether age and sex influence CS isomers and HA in normal synovial fluid, we determined concentrations of chondroitin 6-sulfate (C6S), chondroitin 4-sulfate (C4S), and HA in healthy subjects of different ages. Synovial fluids were obtained from 187 healthy volunteers, 14–89 years of age. Chondroitin 6-sulfate, C4S, HA concentrations, and C6S : C4S ratio showed a significant negative correlation with age. There were no sex-related differences in HA concentration, but the concentrations of C6S and C4S and the C6S : C4S ratio were significantly lower in women than in men in most age groups.