Toxicological evaluation of Yulangsan polysaccharide in Wistar rats: A 26-week oral gavage study

Although numerous studies have proven the medicinal values of Yulangsan polysaccharide (YLSP), the toxicity of this active ingredient is unknown. In the acute toxicity study, a single oral administration of 24 g/kg YLSP caused neither toxicological symptoms nor mortality, and the LD₅₀ was estimated >24 g/kg. In the chronic toxicity study, we administered doses of 0, 0.6, 1.2 and 2.4 g/kg YLSP in rats by oral gavage for 26 weeks followed by a 3-week recovery period. There was no mortality or remarkable clinical signs observed during this 26-week study. Additionally, there were no toxic differences in the following parameters: body weight, food consumption, hematology, clinical biochemistry, organ weight, and macroscopic findings. There were no adverse effects on histopathology observed in males or female rats treated with YLSP. Based on the results, the no-observed-adverse-effect-level of YLSP in rats is greater than 2.4 g/kg when administered orally for 26 consecutive weeks.     

Deficiency of anti-inflammatory cytokine IL-4 leads to neural hyperexcitability and aggravates cerebral ischemia–reperfusion injury

Systematic administration of anti-inflammatory cytokine interleukin 4 (IL-4) has been shown to improve recovery after cerebral ischemic stroke. However, whether IL-4 affects neuronal excitability and how IL-4 improves ischemic injury remain largely unknown. Here we report the neuroprotective role of endogenous IL-4 in focal cerebral ischemia–reperfusion (I/R) injury. In multi-electrode array (MEA) recordings, IL-4 reduces spontaneous firings and network activities of mouse primary cortical neurons. IL-4 mRNA and protein expressions are upregulated after I/R injury. Genetic deletion of Il-4 gene aggravates I/R injury in vivo and exacerbates oxygen-glucose deprivation (OGD) injury in cortical neurons. Conversely, supplemental IL-4 protects Il-4^−/− cortical neurons against OGD injury. Mechanistically, cortical pyramidal and stellate neurons common for ischemic penumbra after I/R injury exhibit intrinsic hyperexcitability and enhanced excitatory synaptic transmissions in Il-4^−/− mice. Furthermore, upregulation of Nav1.1 channel, and downregulations of KCa3.1 channel and α6 subunit of GABA_A receptors are detected in the cortical tissues and primary cortical neurons from Il-4^−/− mice. Taken together, our findings demonstrate that IL-4 deficiency results in neural hyperexcitability and aggravates I/R injury, thus activation of IL-4 signaling may protect the brain against the development of permanent damage and help recover from ischemic injury after stroke.     

华细辛和北细辛HPLC特征图谱识别与抗炎靶点及抗炎成分分析

建立华细辛和北细辛HPLC特征图谱并结合聚类分析研究2种来源细辛的识别方法;应用网络药理学方法预测细辛潜在抗炎靶点并寻找潜在抗炎成分。对89批细辛药材(12批华细辛和77批北细辛)的数据进行分析确定了11个特征峰,用对照品、紫外光谱和LC-MS指认了11个特征成分。特征峰面积聚类分析显示华细辛和北细辛被分为2类,且利用特征峰面积比值可实现两者区分,当特征峰9(细辛素)/参照峰S(卡枯醇)峰面积比值大于5时为华细辛,小于2时为北细辛。对119种细辛成分进行网络药理学分析的结果表明细辛抗炎作用可能与COX-2,COX-1,iNOS,MAPK14,LAT4H,NR3C1,PPARG和TNF等8个靶点相关,其中COX-2最为关键,与5种特征成分细辛脂素、芝麻脂素、细辛素、甲基丁香酚和黄樟醚均存在相互作用。此外,细辛脂素、芝麻脂素与iNOS,MAPK14也存在相互作用关系,黄樟醚和细辛素可作用于iNOS,COX-1,LAT4H,甲基丁香酚可作用于COX-1,LAT4H。细辛脂素与芝麻脂素均可作用在COX-2,iNOS和MAPK143个靶点上,提示它们是细辛发挥抗炎作用的活性成分;COX-2分子对接结果和COX-2活性实验结果验证了细辛脂素、芝麻脂素可抑制COX-2活性,为细辛抗炎作用活性成分。基于HPLC特征图谱的华细辛和北细辛识别方法简便易行;预测到的细辛抗炎靶点和抗炎成分为完善细辛质量评价体系奠定了基础。     

Inactivated influenza vaccine formulated with single-stranded RNA-based adjuvant confers mucosal immunity and cross-protection against influenza virus infection

Influenza vaccination is considered the most valuable means to prevent and control seasonal influenza infections, which causes various clinical symptoms, ranging from mild cough and fever to even death. Among various influenza vaccine types, the inactivated subunit type is known to provide improved safety with reduced reactogenicity. However, there are some drawbacks associated with inactivated subunit type vaccines, with the main ones being its low immunogenicity and the induction of Th2-biased immune responses. In this study, we investigated the role of a single-stranded RNA (ssRNA) derived from the intergenic region in the internal ribosome entry site of the Cricket paralysis virus as an adjuvant rather than the universal vaccine for a seasonal inactivated subunit influenza vaccine. The ssRNA adjuvant stimulated not only well-balanced cellular (indicated by IgG2a, IFN-γ, IL-2, and TNF-α) and humoral (indicated by IgG1 and IL-4) immune responses but also a mucosal immune response (indicated by IgA), a key protector against respiratory virus infections. It also increases the HI titer, the surrogate marker of influenza vaccine efficacy. Furthermore, ssRNA adjuvant confers cross-protective immune responses against heterologous influenza virus infection while promoting enhanced viral clearance. Moreover, ssRNA adjuvant increases the number of memory CD4⁺ and CD8⁺ T cells, which can be expected to induce long-term immune responses. Therefore, this ssRNA-adjuvanted seasonal inactivated subunit influenza vaccine might be the best influenza vaccine generating robust humoral and cellular immune responses and conferring cross-protective and long-term immunity.     

基于UPLC对龙血通络胶囊特征图谱及5个成分含量的测定

目的 建立龙血通络胶囊超高效液相色谱法（UPLC）特征图谱,并测定7,4′-二羟基黄酮、4,4′-二羟基-2,6-二甲氧基二氢查耳酮、龙血素A、龙血素B、紫檀芪5个成分的含量。方法 采用Agilent Infinity Lab Poroshell 120 EC-C₁₈色谱柱（150 mm×3.0 mm,2.7 μm）,以乙腈（A）-0.1%磷酸水溶液（B）为流动相梯度洗脱,柱温为35 ℃,流速为0.5 mL·min^–1,进样体积为2 μL,检测波长为280 nm。结果 建立了龙血通络胶囊UPLC特征图谱,标定了13批样品中的11个共有特征峰,相似度均大于0.95。5个成分在相应质量浓度范围内呈良好的线性关系（r>0.999 5）,加样回收率为97.93%~107.33%,RSD均小于3.0%。结论 建立的UPLC操作简便、高效准确,可为龙血通络胶囊的质量控制提供参考。