Shuxuetong injection composed of leech(Hirudo nipponica Whitman) and earthworm(Pheretima aspergillum) has been used for the clinical treatment of acute stroke for many years in China. However, the precise neuroprotect...Shuxuetong injection composed of leech(Hirudo nipponica Whitman) and earthworm(Pheretima aspergillum) has been used for the clinical treatment of acute stroke for many years in China. However, the precise neuroprotective mechanism of Shuxuetong injection remains poorly understood. Here, cerebral microvascular endothelial cells(bEnd.3) were incubated in glucose-free Dulbecco's modified Eagle's medium containing 95% N_2/5% CO_2 for 6 hours, followed by high-glucose medium containing 95% O_2 and 5% CO_2 for 18 hours to establish an oxygen-glucose deprivation/reperfusion model. This in vitro cell model was administered Shuxuetong injection at 1/32, 1/64, and 1/128 concentrations(diluted 32-, 64-, and 128-times). Cell Counting Kit-8 assay was used to evaluate cell viability. A fluorescence method was used to measure lactate dehydrogenase, and a fluorescence microplate reader used to detect intracellular reactive oxygen species. A fluorescent probe was also used to measure mitochondrial superoxide production. A cell resistance meter was used to measure transepithelial resistance and examine integrity of monolayer cells. The fluorescein isothiocyanate-dextran test was performed to examine blood-brain barrier permeability. Real-time reverse transcription polymerase chain reaction was performed to analyze mRNA expression levels of tumor necrosis factor alpha, interleukin-1β, interleukin-6, and inducible nitric oxide synthase. Western blot assay was performed to analyze expression of caspase-3, intercellular adhesion molecule 1, vascular cell adhesion molecule 1, occludin, vascular endothelial growth factor, cleaved caspase-3, B-cell lymphoma 2, phosphorylated extracellular signal-regulated protein kinase, extracellular signal-regulated protein kinase, nuclear factor-κB p65, I kappa B alpha, phosphorylated I kappa B alpha, I kappa B kinase, phosphorylated I kappa B kinase, claudin-5, and zonula occludens-1. Our results show that Shuxuetong injection increases bEnd.3 cell viability and B-cell lymphoma 2 expression, reduces cleaved caspase-3 expression, inhibits production of reactive oxygen species and mitochondrial superoxide, suppresses expression of tumor necrosis factor alpha, interleukin-1β, interleukin-6, inducible nitric oxide synthase mRNA, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, markedly increases transepithelial resistance, decreases blood-brain barrier permeability, upregulates claudin-5, occludin, and zonula occludens-1 expression, reduces nuclear factor-κB p65 and vascular endothelial growth factor expression, and reduces I kappa B alpha, extracellular signal-regulated protein kinase 1/2, and I kappa B kinase phosphorylation levels. Overall, these findings suggest that Shuxuetong injection has protective effects on brain microvascular endothelial cells after oxygen-glucose deprivation/reperfusion. Moreover, its protective effect is associated with reduction of mitochondrial superoxide production, inhibition of the inflammatory response, and inhibition of vascular endothelial growth factor, extracellular signal-regulated protein kinase 1/2, and the nuclear factor-κB p65 signaling pathway.展开更多
BACKGROUND The first case of pneumonia subsequently attributed to severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) occurred in Wuhan,Hubei Province on December 8,2019.The symptoms included fever,coughing,...BACKGROUND The first case of pneumonia subsequently attributed to severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) occurred in Wuhan,Hubei Province on December 8,2019.The symptoms included fever,coughing,and breathing difficulties.A few patients with this infection may only have atypical symptoms,which could lead to a misdiagnosis and subsequently further facilitate the spread of the virus.CASE SUMMARY A 74-year-old female patient complained of severe diarrhea.She did not have fever,coughing,or breathing difficulties.A physical examination revealed no obvious positive signs.The patient had been hypertensive for more than 10 years.Her blood pressture was well controlled.On January 9,2020,the patient’s son visited a colleague who was later confirmed positive for SARS-CoV-2 and his first close contact with our patient was on January 17.The patient was first diagnosed with gastrointestinal dysfunction.However,considering her indirect contact with a SARS-CoV-2-infected individual,we suggested that an atypical pneumonia virus infection should be ruled out.A computed tomography scan was performed on January 26,and showed ground-glass nodules scattered along the two lungs,suggestive of viral pneumonia.Given the clinical characteristics,epidemiological history,and examination,the patient was diagnosed with corona virus disease-2019 (COVID-19).CONCLUSION Our patient had atypical symptoms of COVID-19.Careful acquisition of an epidemiological history is necessary to make a correct diagnosis and strategize a treatment plan.展开更多
Objective: To explore the effect of NBP on the serum inflammatory cytokines, oxidative stress, and NF in patients with acute cerebral infarction (ACI). Methods: A total of 118 patients with ACI who were admitted in ou...Objective: To explore the effect of NBP on the serum inflammatory cytokines, oxidative stress, and NF in patients with acute cerebral infarction (ACI). Methods: A total of 118 patients with ACI who were admitted in our hospital from October 2016 to September 2017 were included in the study and randomized into the observation group and the control group, with 59 cases in each group. The patients in the control group were given blood pressure control, blood sugar control, micro-circulation improvement, anti-platelet aggregation, nerve nourishing, and lipid regulation. On the above basis, the patients in the observation group were given intravenous injection of NBP sodium chloride injection. The patients in the two groups were treated for 14 d. The morning fasting venous blood before treatment, 7 d and 14 d after treatment in the two groups was collected, and centrifuged for the serum. And IL-6, TFN-α, hs-CRP, SOD, MDA, NSE, VEGF, BDNF, and bFGF were measured. Results: 7 d and 14 d after treatment, the serum IL-6, TFN-α and hs-CRP in the observation group were significantly lower than those in the control group. Besides, 7 d and 14 d after treatment, the serum SOD in the observation group was significantly higher than that in the control group, while MDA was significantly lower than that in the control group. The serum NSE 7 d and 14 d after treatment in the observation group was significantly lower than that in the control group, while VEGF, BDNF, and bFGF were significantly higher than those in the control group. Conclusions: NBP in the treatment of ACI can down regulate the expression of inflammatory cytokines to play an anti-inflammatory effect, increase the antioxidant ability to remove OFR, improve the neurocyte metabolism, and effectively play the nerve protecting and prognosis improvement role.展开更多
Objective： Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion （I/R） injury, resulting in neuronal death. Salvianolate Lyophili...Objective： Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion （I/R） injury, resulting in neuronal death. Salvianolate Lyophilized Injection （SLI） and Xueshuantong Injection （Lyophilized） （XST）, which have been widely used in the treatment of acutely cerebral infarction clinically in China, exhibit various biological activities. In this study, the neuroprotective properties of SLI combined with XST in a rat model of middle cerebral artery occlusion- reperfusion （MCAO/R） were investigated. Methods： In this study, male Wistar rats were subjected to 1.5h of middle cerebral artery occlusion followed by reperfusion for 24 h. The rats were randomly divided into the following six groups： normal group （NOR）, model group （MOD）, SLI group （21 mg/kg, SLI）, ）（ST group （100 mg/kg, ）（ST）, SLI combined with XST （XST 100 mg/kg ＋ SLI 21 mg/kg, 1X1S）, and Edaravone （as a positive control drug, 6 mL/kg, EDI）, once a day for 3 d. The neuronal injury, the expression of glial fibrillary acidic protein （GFAP） and ionized calcium binding adaptor molecule 1 （IBA-1）, and the changes of pro-inflammatory mediators interleukin- 6 （IL-6）, tumor necrosis factor alpha （TNF-α） and anti-inflammatory mediator interleukin-10 （IL-10） were observed. Results： 1X1S treatment significantly increased the number of neuron, compared with the MOD group, SH group and XST group. Gliosis （GFAP and IBA-1） and expression of pro-inflammatory mediators IL-6 and TNF-a were significantly reduced. Meanwhile, 1XIS significantly increased the expression of anti- inflammatory mediator IL-10 in the brains of MCAO/R rats, compared with the MOD group, SLI and XST groups. SLI and XST also remarkably down-regulated the expression of IL-6 and TNF-α compared with the MOD group. Conclusions： This study shows that SLI combined with XST （1X1S） can protect cerebral ischemia- reperfusion injury due to its anti-inflammatory property, and may provide a potential promising new therapeutic strategy for acute ischemic stroke.展开更多
基金supported in part by the National Natural Science Foundation of China,No.81573644(to LMH),81573733(to SWX)the Tianjin 131 Innovative Team Project,China(to HW)+5 种基金the National Major Science and Technology Project of China,No.2012ZX09101201-004(to SWX)the Science and Technology Plan Project of Tianjin of China,No.16PTSYJC00120(to LMH)the Applied Foundation and Frontier Technology Research Program of Tianjin of China(General Project),No.14JCYBJC28900(to SXW)the National International Science and Technology Cooperation Project of China,No.2015DFA30430(to HW)the Key Program of the Natural Science Foundation of Tianjin of China,No.16ICZDJC36300(to HW)the Scientific Research and Technology Development Plan Project of Guangxi Zhuang Autonomous Region of China,No.14125008-2-5(to SXW)
文摘Shuxuetong injection composed of leech(Hirudo nipponica Whitman) and earthworm(Pheretima aspergillum) has been used for the clinical treatment of acute stroke for many years in China. However, the precise neuroprotective mechanism of Shuxuetong injection remains poorly understood. Here, cerebral microvascular endothelial cells(bEnd.3) were incubated in glucose-free Dulbecco's modified Eagle's medium containing 95% N_2/5% CO_2 for 6 hours, followed by high-glucose medium containing 95% O_2 and 5% CO_2 for 18 hours to establish an oxygen-glucose deprivation/reperfusion model. This in vitro cell model was administered Shuxuetong injection at 1/32, 1/64, and 1/128 concentrations(diluted 32-, 64-, and 128-times). Cell Counting Kit-8 assay was used to evaluate cell viability. A fluorescence method was used to measure lactate dehydrogenase, and a fluorescence microplate reader used to detect intracellular reactive oxygen species. A fluorescent probe was also used to measure mitochondrial superoxide production. A cell resistance meter was used to measure transepithelial resistance and examine integrity of monolayer cells. The fluorescein isothiocyanate-dextran test was performed to examine blood-brain barrier permeability. Real-time reverse transcription polymerase chain reaction was performed to analyze mRNA expression levels of tumor necrosis factor alpha, interleukin-1β, interleukin-6, and inducible nitric oxide synthase. Western blot assay was performed to analyze expression of caspase-3, intercellular adhesion molecule 1, vascular cell adhesion molecule 1, occludin, vascular endothelial growth factor, cleaved caspase-3, B-cell lymphoma 2, phosphorylated extracellular signal-regulated protein kinase, extracellular signal-regulated protein kinase, nuclear factor-κB p65, I kappa B alpha, phosphorylated I kappa B alpha, I kappa B kinase, phosphorylated I kappa B kinase, claudin-5, and zonula occludens-1. Our results show that Shuxuetong injection increases bEnd.3 cell viability and B-cell lymphoma 2 expression, reduces cleaved caspase-3 expression, inhibits production of reactive oxygen species and mitochondrial superoxide, suppresses expression of tumor necrosis factor alpha, interleukin-1β, interleukin-6, inducible nitric oxide synthase mRNA, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, markedly increases transepithelial resistance, decreases blood-brain barrier permeability, upregulates claudin-5, occludin, and zonula occludens-1 expression, reduces nuclear factor-κB p65 and vascular endothelial growth factor expression, and reduces I kappa B alpha, extracellular signal-regulated protein kinase 1/2, and I kappa B kinase phosphorylation levels. Overall, these findings suggest that Shuxuetong injection has protective effects on brain microvascular endothelial cells after oxygen-glucose deprivation/reperfusion. Moreover, its protective effect is associated with reduction of mitochondrial superoxide production, inhibition of the inflammatory response, and inhibition of vascular endothelial growth factor, extracellular signal-regulated protein kinase 1/2, and the nuclear factor-κB p65 signaling pathway.
基金the National Natural Science Foundation of China,No.81801379Top-level Clinical Discipline Project of Shanghai Pudong District,No.PWYgf2018-02.
文摘BACKGROUND The first case of pneumonia subsequently attributed to severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) occurred in Wuhan,Hubei Province on December 8,2019.The symptoms included fever,coughing,and breathing difficulties.A few patients with this infection may only have atypical symptoms,which could lead to a misdiagnosis and subsequently further facilitate the spread of the virus.CASE SUMMARY A 74-year-old female patient complained of severe diarrhea.She did not have fever,coughing,or breathing difficulties.A physical examination revealed no obvious positive signs.The patient had been hypertensive for more than 10 years.Her blood pressture was well controlled.On January 9,2020,the patient’s son visited a colleague who was later confirmed positive for SARS-CoV-2 and his first close contact with our patient was on January 17.The patient was first diagnosed with gastrointestinal dysfunction.However,considering her indirect contact with a SARS-CoV-2-infected individual,we suggested that an atypical pneumonia virus infection should be ruled out.A computed tomography scan was performed on January 26,and showed ground-glass nodules scattered along the two lungs,suggestive of viral pneumonia.Given the clinical characteristics,epidemiological history,and examination,the patient was diagnosed with corona virus disease-2019 (COVID-19).CONCLUSION Our patient had atypical symptoms of COVID-19.Careful acquisition of an epidemiological history is necessary to make a correct diagnosis and strategize a treatment plan.
文摘Objective: To explore the effect of NBP on the serum inflammatory cytokines, oxidative stress, and NF in patients with acute cerebral infarction (ACI). Methods: A total of 118 patients with ACI who were admitted in our hospital from October 2016 to September 2017 were included in the study and randomized into the observation group and the control group, with 59 cases in each group. The patients in the control group were given blood pressure control, blood sugar control, micro-circulation improvement, anti-platelet aggregation, nerve nourishing, and lipid regulation. On the above basis, the patients in the observation group were given intravenous injection of NBP sodium chloride injection. The patients in the two groups were treated for 14 d. The morning fasting venous blood before treatment, 7 d and 14 d after treatment in the two groups was collected, and centrifuged for the serum. And IL-6, TFN-α, hs-CRP, SOD, MDA, NSE, VEGF, BDNF, and bFGF were measured. Results: 7 d and 14 d after treatment, the serum IL-6, TFN-α and hs-CRP in the observation group were significantly lower than those in the control group. Besides, 7 d and 14 d after treatment, the serum SOD in the observation group was significantly higher than that in the control group, while MDA was significantly lower than that in the control group. The serum NSE 7 d and 14 d after treatment in the observation group was significantly lower than that in the control group, while VEGF, BDNF, and bFGF were significantly higher than those in the control group. Conclusions: NBP in the treatment of ACI can down regulate the expression of inflammatory cytokines to play an anti-inflammatory effect, increase the antioxidant ability to remove OFR, improve the neurocyte metabolism, and effectively play the nerve protecting and prognosis improvement role.
基金financially supported by the National Natural Science Foundation (81573644)the Natural Science Foundation of Tianjin (14JCYBJC28900)the Tianjin Technology Innovation System and the Condition of Platform Construction Plan (16PTSYJC00120)
文摘Objective： Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion （I/R） injury, resulting in neuronal death. Salvianolate Lyophilized Injection （SLI） and Xueshuantong Injection （Lyophilized） （XST）, which have been widely used in the treatment of acutely cerebral infarction clinically in China, exhibit various biological activities. In this study, the neuroprotective properties of SLI combined with XST in a rat model of middle cerebral artery occlusion- reperfusion （MCAO/R） were investigated. Methods： In this study, male Wistar rats were subjected to 1.5h of middle cerebral artery occlusion followed by reperfusion for 24 h. The rats were randomly divided into the following six groups： normal group （NOR）, model group （MOD）, SLI group （21 mg/kg, SLI）, ）（ST group （100 mg/kg, ）（ST）, SLI combined with XST （XST 100 mg/kg ＋ SLI 21 mg/kg, 1X1S）, and Edaravone （as a positive control drug, 6 mL/kg, EDI）, once a day for 3 d. The neuronal injury, the expression of glial fibrillary acidic protein （GFAP） and ionized calcium binding adaptor molecule 1 （IBA-1）, and the changes of pro-inflammatory mediators interleukin- 6 （IL-6）, tumor necrosis factor alpha （TNF-α） and anti-inflammatory mediator interleukin-10 （IL-10） were observed. Results： 1X1S treatment significantly increased the number of neuron, compared with the MOD group, SH group and XST group. Gliosis （GFAP and IBA-1） and expression of pro-inflammatory mediators IL-6 and TNF-a were significantly reduced. Meanwhile, 1XIS significantly increased the expression of anti- inflammatory mediator IL-10 in the brains of MCAO/R rats, compared with the MOD group, SLI and XST groups. SLI and XST also remarkably down-regulated the expression of IL-6 and TNF-α compared with the MOD group. Conclusions： This study shows that SLI combined with XST （1X1S） can protect cerebral ischemia- reperfusion injury due to its anti-inflammatory property, and may provide a potential promising new therapeutic strategy for acute ischemic stroke.
