核因子κB受体活化因子配体和肿瘤坏死因子α经炎性牙周膜干细胞外泌体促进破骨细胞分化

目的　慢性牙周炎表现的病理性骨吸收非常常见，牙周膜干细胞（PDLSCs）的外泌体（Exo）对骨吸收的作用和影响尚不明确，本研究分析了该Exo蛋白组分对破骨细胞分化的影响。方法　分别从正畸前拔牙患者和慢性牙周炎患者的牙周膜组织中提取PDLSCs，通过流式细胞术检测表面标记物；通过差速离心分别提取2种细胞的Exo，即Exo-WT和Exo-CP，并通过Western blot、透射电子显微镜（TEM）、蛋白浓度检测、纳米粒径追踪检测Exo特征；通过蛋白质谱检测2种Exo的蛋白组分；通过酶联免疫吸附（ELISA）验证了差异表达蛋白肿瘤坏死因子α（TNF-α）、核因子κB受体活化因子配体（RANKL）、白细胞介素（IL）-1α、转化生长因子β（TGF-β）、骨形态发生蛋白2（BMP-2）表达水平；将10、100、1 000 μg·mL^-1的Exo-CP或Exo-WT加入RAW264.7培养基中并于5 d时通过实时荧光定量聚合酶链反应（RT-qPCR）、Western blot、抗酒石酸酸性磷酸酶（TRAP）染色检测破骨分化相关指标表达情况；采用SPSS 24.0软件对实验数据进行统计学分析。结果　Exo-CP富集的差异表达蛋白主要与破骨细胞分化的TNF信号通路、核转录因子κB（NF-κB）信号通路相关；ELISA实验证实了Exo-CP中高表达TNF-α、RANKL、IL-1α，低表达TGF-β1、BMP-2（P<0.05）；Exo-CP作用于RAW264.7，显著提高了细胞的破骨分化相关基因及蛋白的表达水平，TRAP染色可见分化的破骨细胞，且呈现浓度依赖性，100、1 000 μg·mL^-1浓度Exo-CP对破骨细胞分化的促进作用显著高于10 μg·mL^-1浓度组（P<0.05）。结论　慢性牙周炎的病理性骨吸收可能由炎性PDLSCs所分泌的Exo通过促进破骨细胞分化引起，Exo中主要的作用蛋白可能为RANKL和TNF-α，本研究为慢性牙周炎骨吸收的发病机制提供了新视角。     

与结核相关的脂膜炎研究进展

【摘要】 脂膜炎病因繁多而复杂，感染为重要病因之一，在欧美等结核低发区，以链球菌感染最为常见，但在结核高发地区如我国，结核杆菌感染是脂膜炎的重要因素，以Bazin硬红斑和结节性红斑最为常见。Bazin硬红斑与结核关系最为密切，常表现为下肢屈侧结节、斑块，可出现溃疡，病理表现为小叶性脂膜炎。结节性红斑可作为结核的早期症状出现，可作为结核的预测指标，常表现为双下肢伸侧红斑、结节，极少破溃，间隔性脂膜炎是其病理特征。Bazin硬红斑和结节性红斑的抗结核治疗常可取得理想效果。因此，明确结核分支杆菌与脂膜炎的关系，在脂膜炎的治疗及预后方面，尤其在结核高发地区有着重要意义。     

基于ArcCHECK-3DVH系统的鼻咽癌容积旋转调强放疗三维剂量验证研究

目的:探讨ArcCHECK-3DVH系统在鼻咽癌容积旋转调强放疗（VMAT）三维剂量验证中的应用。方法:选取基于Monaco治疗计划系统（TPS）制定的20例鼻咽癌患者VMAT计划，利用ArcCHECK-3DVH系统进行患者计划的验证。比较TPS计算剂量与ArcCHECK-3DVH系统基于测量重建剂量的三维γ通过率（3%/3 mm, TH=10%），以及靶区剂量参数[D98%]、[D2%]和[Dmean]与危及器官剂量参数[Dmax]和[Dmean]等。结果:患者计划验证的三维γ通过率为98.20%（97.75%, 99.20%），TPS计算靶区内剂量和ArcCHECK-3DVH重建的剂量在[D98%]、[D2%]和[Dmean]参数上的差异值大部分小于3%，极个别情况的最大差异值在5%以内，绝大部分患者除PGTVnd外的各个靶区γ通过率为90%以上；各个危及器官对应的[Dmax]和[Dmean]剂量参数差异值均小于6%，γ通过率均在93%以上。结论:ArcCHECK-3DVH系统重建的剂量结果与TPS计算结果符合较好，为鼻咽癌VMAT计划剂量验证提供了丰富的剂量参数信息，有助于患者治疗计划的质量验证。     

Treponemal antibody in CSF and cellular immunity in peripheral blood of syphilitic patients with persisting positive rapid plasma regain

The ratio of patients with RPR constant positive more than 2 years despite receiving standard syphilis treatment has been reported to be 11.54%~31.3%. The current interpretations on this phenomenon are cellular immune function restrained and the existence of neurosyphilis or asymptomatic neurosyphilis. We conducted this study to detect the treponemal antibody in cerebrospinal fluid (CSF) and lymphocyte subsets in peripheral blood of syphilis patients with persisting RPR positive more than 2 years without neurologic signs, and then explore their relationship. In this study, Treponemal antibody in CSF of 46 syphilitic with HIV negative were measured by syphilis serum test and compared with that of 5 neurosyphilis. Lymphocyte subsets were measured by flow cytometry (FCM) and compared with that of 30 healthy controls. We observed that treponemal antibody in CSF was detected not only in 12 cases (25.21%) of 46 treated patients, but also in 5 neurosyphilis. The ratio of lymphocyte subsets revealed that CD3⁺, CD4⁺ T cells and natural killer (NK) cells showed no significant differences between the patient and healthy controls (P > 0.05), while CD8⁺ T cells in patients were significant higher than that in healthy controls (P < 0.001). Lymphocyte subsets showed no significant differences between the patients with treponemal antibody positive and negative in CSF (P > 0.05). In conclusion, the treponemal antibody in CSF of treated patients suggests that part of them were asymptomatic neurosyphilis and with cellular immunodifeciency. And there is no significant relationship between asymptomatic neurosyphilis and cellular immunodeficiency in peripheral blood.     

Pharmacokinetics of oral and intravenous cannabidiol and its antidepressant-like effects in chronic mild stress mouse model

Cannabidiol (CBD) exhibits significant efficacy in mental and inflammatory diseases. Several studies have recently reported on the rapid antidepressant-like effects of CBD, suggesting that CBD is a potential anti-depressant or anti-stress drug. However, CBD is mainly administered orally or by inhalation with poor bioavailability, resulting in high costs. We aim to explore the efficacy of long-term periodic administration of CBD in chronic mild stress (CMS) via two routes and its pharmacokinetics. We treated ICR mice with CBD administered orally and intravenously and then determined the kinetic constants. A single bolus intravenous injection of CBD resulted in a half-life of 3.9 h, mean residence time of 3.3 h, and oral bioavailability of about 8.6%. The antidepressant-like effects of periodically administered CBD on the chronic mild stress mouse model are evaluated. Results demonstrated that such treatment at a high dose of 100 mg/kg CBD (p.o.) or a low dose of 10 mg/kg CBD (i.v.), elicited significant antidepressant-like behavioral effects in forced swim test, following increased mRNA expression of brain-derived neurotrophic factor (BDNF) and synaptophysin in the prefrontal cortex and the hippocampus. Our findings are expected to provide a reference for the development of intravenous antidepressant formulations of CBD.     

网络药理学及分子对接法探讨三仁汤主要成分对新型冠状病毒肺炎（COVID-19）的机制研究

目的通过网络药理学及分子对接法探讨三仁汤主要成分对新型冠状病毒肺炎(COVID-19)的作用机制。方法通过中药系统药理学分析平台(TCMSP)及中医整合药理学研究平台V2.0(TCMIP V2.0)获得三仁汤君药(苦杏仁、白蔻仁、薏苡仁)全部化学成分并筛选,结合相关文献筛得主要活性成分。借助Uni Port数据库查询活性成分对应靶标蛋白,Cytoscape3.7.2构建成分-靶点网络。通过STRING网站构建PPI蛋白互作网络,运用cytoHubba分析关键子网络。CTD数据库对三仁汤君药活性成分靶蛋白进行GO、KEGG富集分析。以三仁汤君药活性成分及洛匹那韦等相关化学药物为配体,通过CB-Dock网站分别与新型冠状病毒(SARS-CoV-2)3CL水解酶进行分子对接。结果三仁汤君药共有39个活性成分,对应有效靶标168个。GO功能富集分析筛得生物学过程(BP)、细胞组成(CC)、分子功能(MF)分别为25条、14条及2条。KEGG富集筛得先天免疫系统(innate immune system)、免疫系统中的细胞因子信号转导(cytokine signaling in immune system)及白介素信号通路(signaling by interleukins)等信号通路,共36条。分子对接提示三仁汤君药活性成分白桦脂酸、枣素及齐墩果酸与SARS-CoV-2 3CL水解酶结合性较好,三者的Vina得分与3CLpro抑制剂洛匹那韦及Rd Rp抑制剂瑞德西韦的得分相近。结论三仁汤君药可能通过调控炎症因子,参与炎症相关信号通路,以多成分、多靶点、多途径参与COVID-19的多种生理过程,对治疗COVID-19发挥一定的干预作用,且其活性成分具有潜在抗SARS-CoV-2的可能性。     

麻风病患者抑郁状态与生存质量关系的多重线性回归分析

目的 分析麻风病患者抑郁状态与生存质量各维度间的相关关系。方法 采用流行病调查用抑郁自评量表（center for epidemiologic studies depression scale,CES-D）和世界卫生组织生存质量简表（world health organization's quality of life questionnaire-brief version,WHOQOL-bref）对广东省麻风病患者进行抑郁程度和生存质量调查,用多重线性回归方法分析抑郁程度得分与生存质量各维度间的关系和强度。结果 共263名麻风病患者完成了调查,其中男性177名（67.3%）,女性86名（32.7%）。131名（49.8%）调查对象有抑郁症状,有抑郁症状调查对象生存质量各维度评分均低于无抑郁症状对象,差异均具有统计学意义（均有P<0.05）。在各维度中,生存质量总评、生理健康、心理健康、社会关系和环境因素得分与抑郁症状的回归系数分别为-0.340、-0.322、-0.644、-0.424和0.244,除生存质量总评外,其他回归系数差异均具有统计学意义（均有P<0.05）。结论 广东省麻风病患者的抑郁症状发生率高,生存质量得分与CES-D得分有关联,应采用多层次的干预措施防止麻风病患者抑郁症的发生。     

Involvement of microglial activation in the brainstem in experimental dental injury and inflammation

Objective

The purpose of this study was to determine the effect of dental injury and inflammation on microglia in the trigeminal subnucleus caudalis (Vc).

Methods

Pulp exposure (PX) was performed on the first maxillary molar of 35 rats. Specimens were collected at 1, 3, 7, 14, 21 and 28 days after PX. Teeth were processed for H&E staining and immunohistochemical staining for OX-42, a marker of microgial activation, in the Vc.

Results

We observed that there was a progressive and persistent inflammation in the tooth. At 21-28 days after PX, the inflammation extended out into periodontal ligament. Simultaneously, significant microglial activation was observed which starting at 2 weeks and peaking at 4 weeks.

Conclusion

Dental injury and inflammation induced microglial activation in the Vc. The results indicate that activation of microglia may be implicated in the central mechanisms of pain that can be associated with dental inflammation.     

一致性评价系数应用于无序多分类资料的效果评价

目的 基于AC1系数的构建思想,建立一致性评价系数（CEA）在两评价者无序多分类结局的评价方法,并可避免Kappa系数的缺陷。方法 通过随机抽样生成诊断试验类型数据,使用Monte Carlo模拟,在样本量、指定事件在总体的占比、偶然评价率、类别数等不同参数组合下进行重抽样,比较Kappa系数、AC1系数和CEA系数的均方误、方差和方差的期望。通过从总体中随机抽样1000次得到CEA系数的分布描述。结果 偶然评价率的不一致会导致CEA系数的均方误波动较大。与Kappa系数相比,AC1系数和CEA系数在指定事件的占比为极端值的情况更为稳定。在小样本、偶然评价率不一致的情况下,Kappa系数的方差和方差的期望变大,CEA系数变化较小。大样本条件下,CEA系数近似服从正态分布。结论 Kappa系数、AC1系数、CEA系数均受偶然评价率的影响最大,样本量次之。针对无序多分类结局,CEA系数在不同的样本量、偶然评价率下具有更稳健的性质。