建立了有效分离重组人胰岛素注射液中B3和B3iso脱氨人胰岛素的反相高效液相色谱(RP-HPLC)法,并采用高效液相色谱-四极杆-飞行时间质谱联用技术(HPLC-MS/MS)对其进行氨基酸序列的鉴定。在《中国药典》2010版重组人胰岛素注射液有关物质...建立了有效分离重组人胰岛素注射液中B3和B3iso脱氨人胰岛素的反相高效液相色谱(RP-HPLC)法,并采用高效液相色谱-四极杆-飞行时间质谱联用技术(HPLC-MS/MS)对其进行氨基酸序列的鉴定。在《中国药典》2010版重组人胰岛素注射液有关物质检查分析方法的基础上,优化流动相p H值,以0.2mol/L硫酸盐缓冲液(p H 3.6)-乙腈(90∶10)作为A相,乙腈-水(50∶50)为B相,梯度洗脱,分别收集杂质峰1和杂质峰2,脱盐浓缩后,用丝氨酸蛋白酶(V8酶)酶切,取部分酶切溶液经三(2-羧乙基)膦盐酸盐(TCEP)还原,分别得到还原和非还原肽段溶液,利用LC-MS/MS鉴定氨基酸序列,未知杂质分别为重组人胰岛素B3位发生脱氨和异构化的产物,即B3和B3iso脱氨人胰岛素。将优化后的方法与《中国药典》方法对市售重组人胰岛素注射液进行有关物质的对比分析,已上市厂家的重组人胰岛素注射液均有不同程度的B3和B3iso脱氨人胰岛素存在。该方法不影响重组人胰岛素其它有关物质的检测,可用于监控人胰岛素注射液产品储存过程中脱氨类杂质的增长趋势。展开更多
Dual-frequency satellite positioning receivers are widely used because they can eliminate ionospheric delay and solve the full-circumference ambiguity quickly.However,in traditional dual-frequency receivers,the releva...Dual-frequency satellite positioning receivers are widely used because they can eliminate ionospheric delay and solve the full-circumference ambiguity quickly.However,in traditional dual-frequency receivers,the relevance of dual-frequency signals are not considered,and,with no improvement imposed to the tracking loop,two independent tracking loops are used to achieve the tracking of dual-frequency signals.In this paper,the Bei Dou dual-frequency signals joint tracking algorithm based on Kalman filter is proposed for the tracking of Bei Dou B1I and B3I dual-frequency signals.Taking the relevance of B1I and B3I signals into consideration,the algorithm adds a Kalman filter between the phase detector and carrier loop filter of the traditional dual-frequency independent tracking loop.The output results of the phase detectors of the B1I and B3I branches are then combined and filtered by the Kalman filter,and the results are input to the carrier loop filters of the corresponding branches.Proved by experiments,the algorithm not only enables the loop to enter a stable tracking state quickly,but also reduces the noise bandwidth of the two loop filters by about 10 Hz with the same tracking performance obtained.展开更多
目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化...目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化浊解毒方高剂量组(高剂量组)、益气活血化浊解毒方高剂量+2,5-己酮可可碱(DMXAA)组(高剂量+DMXAA组),每组15只。另取15只正常大鼠为Sham组。低剂量组大鼠灌胃0.2 mL 27.5 g/mL的益气活血化浊解毒方药液,高剂量组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液,高剂量+DMXAA组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液和25 mg/kg的DMXAA,Sham组和缺血性脑卒中组给予等体积生理盐水代替药物;除高剂量+DMXAA组外,其余组大鼠再给予等体积二甲基亚砜溶液灌胃,1 d/次,连续4周。Zea-Longa评分和网屏试验评分评估大鼠神经功能和神经行为学;检测大鼠脑组织含水量;TTC染色法检测大鼠脑梗死面积;酶联免疫吸附试验检测大鼠血清LDH和NGF水平和缺血侧皮层组织TNF-α、IL-6、IL-10水平;HE染色观察大鼠缺血侧皮层组织病理变化;Western blotting检测大鼠缺血侧皮层组织iNOS、Iba1、CD68、CD40、CD206、Arg-1、Cleaved caspase-3、KAT3B、STING蛋白表达。结果缺血性脑卒中组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于Sham组(P<0.05),低剂量组和高剂量组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均低于缺血性脑卒中组(P<0.05),高剂量+DMXAA组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于高剂量组(P<0.05)。缺血性脑卒中组大鼠NGF和IL-10水平低于Sham组,LDH、TNF-α和IL-6水平高于Sham组(P<0.05);低剂量组和高剂量组大鼠NGF和IL-10水平高于缺血性脑卒中组,LDH、TNF-α和IL-6水平低于缺血性脑卒中组(P<0.05);高剂量+DMXAA组大鼠NGF和IL-10水平低于高剂量组,LDH、TNF-α和IL-6高于高剂量组(P<0.05)。与缺血性脑卒中组比较,低剂量组和高剂量组缺血侧皮层神经元形态有明显改善。缺血性脑卒中组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比Sham组高,CD206和Arg-1蛋白相对表达量比Sham组低(P<0.05);低剂量组和高剂量组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比缺血性脑卒中组低,CD206和Arg-1蛋白相对表达量比缺血性脑卒中组高(P<0.05);高剂量+DMXAA组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比高剂量组高,CD206和Arg-1蛋白相对表达量比高剂量组低(P<0.05)。结论益气活血化浊解毒方可调节缺血性脑卒中大鼠小胶质细胞极化,减轻神经炎症和神经损伤,改善神经功能,可能与抑制KAT3B/STING轴有关。展开更多
文摘建立了有效分离重组人胰岛素注射液中B3和B3iso脱氨人胰岛素的反相高效液相色谱(RP-HPLC)法,并采用高效液相色谱-四极杆-飞行时间质谱联用技术(HPLC-MS/MS)对其进行氨基酸序列的鉴定。在《中国药典》2010版重组人胰岛素注射液有关物质检查分析方法的基础上,优化流动相p H值,以0.2mol/L硫酸盐缓冲液(p H 3.6)-乙腈(90∶10)作为A相,乙腈-水(50∶50)为B相,梯度洗脱,分别收集杂质峰1和杂质峰2,脱盐浓缩后,用丝氨酸蛋白酶(V8酶)酶切,取部分酶切溶液经三(2-羧乙基)膦盐酸盐(TCEP)还原,分别得到还原和非还原肽段溶液,利用LC-MS/MS鉴定氨基酸序列,未知杂质分别为重组人胰岛素B3位发生脱氨和异构化的产物,即B3和B3iso脱氨人胰岛素。将优化后的方法与《中国药典》方法对市售重组人胰岛素注射液进行有关物质的对比分析,已上市厂家的重组人胰岛素注射液均有不同程度的B3和B3iso脱氨人胰岛素存在。该方法不影响重组人胰岛素其它有关物质的检测,可用于监控人胰岛素注射液产品储存过程中脱氨类杂质的增长趋势。
基金supported by the National Natural Science Foundation of China (No.51505221)the Nanjing University of Aeronautics and Astronautics Graduate Innovation Base (Lab) Open Fund (No.kfjj20190312)
文摘Dual-frequency satellite positioning receivers are widely used because they can eliminate ionospheric delay and solve the full-circumference ambiguity quickly.However,in traditional dual-frequency receivers,the relevance of dual-frequency signals are not considered,and,with no improvement imposed to the tracking loop,two independent tracking loops are used to achieve the tracking of dual-frequency signals.In this paper,the Bei Dou dual-frequency signals joint tracking algorithm based on Kalman filter is proposed for the tracking of Bei Dou B1I and B3I dual-frequency signals.Taking the relevance of B1I and B3I signals into consideration,the algorithm adds a Kalman filter between the phase detector and carrier loop filter of the traditional dual-frequency independent tracking loop.The output results of the phase detectors of the B1I and B3I branches are then combined and filtered by the Kalman filter,and the results are input to the carrier loop filters of the corresponding branches.Proved by experiments,the algorithm not only enables the loop to enter a stable tracking state quickly,but also reduces the noise bandwidth of the two loop filters by about 10 Hz with the same tracking performance obtained.
文摘目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化浊解毒方高剂量组(高剂量组)、益气活血化浊解毒方高剂量+2,5-己酮可可碱(DMXAA)组(高剂量+DMXAA组),每组15只。另取15只正常大鼠为Sham组。低剂量组大鼠灌胃0.2 mL 27.5 g/mL的益气活血化浊解毒方药液,高剂量组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液,高剂量+DMXAA组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液和25 mg/kg的DMXAA,Sham组和缺血性脑卒中组给予等体积生理盐水代替药物;除高剂量+DMXAA组外,其余组大鼠再给予等体积二甲基亚砜溶液灌胃,1 d/次,连续4周。Zea-Longa评分和网屏试验评分评估大鼠神经功能和神经行为学;检测大鼠脑组织含水量;TTC染色法检测大鼠脑梗死面积;酶联免疫吸附试验检测大鼠血清LDH和NGF水平和缺血侧皮层组织TNF-α、IL-6、IL-10水平;HE染色观察大鼠缺血侧皮层组织病理变化;Western blotting检测大鼠缺血侧皮层组织iNOS、Iba1、CD68、CD40、CD206、Arg-1、Cleaved caspase-3、KAT3B、STING蛋白表达。结果缺血性脑卒中组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于Sham组(P<0.05),低剂量组和高剂量组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均低于缺血性脑卒中组(P<0.05),高剂量+DMXAA组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于高剂量组(P<0.05)。缺血性脑卒中组大鼠NGF和IL-10水平低于Sham组,LDH、TNF-α和IL-6水平高于Sham组(P<0.05);低剂量组和高剂量组大鼠NGF和IL-10水平高于缺血性脑卒中组,LDH、TNF-α和IL-6水平低于缺血性脑卒中组(P<0.05);高剂量+DMXAA组大鼠NGF和IL-10水平低于高剂量组,LDH、TNF-α和IL-6高于高剂量组(P<0.05)。与缺血性脑卒中组比较,低剂量组和高剂量组缺血侧皮层神经元形态有明显改善。缺血性脑卒中组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比Sham组高,CD206和Arg-1蛋白相对表达量比Sham组低(P<0.05);低剂量组和高剂量组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比缺血性脑卒中组低,CD206和Arg-1蛋白相对表达量比缺血性脑卒中组高(P<0.05);高剂量+DMXAA组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比高剂量组高,CD206和Arg-1蛋白相对表达量比高剂量组低(P<0.05)。结论益气活血化浊解毒方可调节缺血性脑卒中大鼠小胶质细胞极化,减轻神经炎症和神经损伤,改善神经功能,可能与抑制KAT3B/STING轴有关。
