Effect of Qingkailing injection on platelets function preserved in vitro / 中国输血杂志

【Objective】 To investigate the effect of Qingkailing injection on platelet function preserved in vitro. 【Methods】 A total of 15 bags of plateletpheresis (≥250 mL/bag), adding Qingkailing injection(1.25 mL/bag) at the final concentration of 1%, were stored at 22 ℃ with gentle agitation as the experimental group, and samples were collected on day 1, 3, 5, 8, 10 and 14 to detect the thromboelastogram (TEG), CD62p expression rate and mitochondrial membrane potential (JC-1). The control group was set up synchronously, with the same volume and storage conditions as the experimental groups, and samples were taken on day 1, 3 and 5 after preservation to undertake the same test items as the experimental groups. The differences of detection indexes between the two groups were compared. 【Results】 1) In the experimental group, there was no significant change in K and α during day 1 to 14(P>0.05). The R value (min) increased from 6.12±1.58 to 11.02±2.26, and the CI value changed from 0.27±1.24 to -10.47±3.51 (P0.05), but decreased to 53.18±2.71 on day 8 and 22.88±3.45 on day 14 (P0.05), but JC-1 (%) was 86.75±3.88 vs 70.36±19.8 on day 5 (P<0.05). In the experimental group, JC-1(%) was 81.04±9.50 vs 71.38±8.77 vs 82.77±7.17 on day 8, 10 and 14, respectively. 【Conclusion】 The activation and aggregation as well as anti-apoptosis function of plateletpheresis, adding Qingkailing injection at the final concentration of 1%, are similar to those of routine preservation.

[Study on synergistic effect of Qingkailing Injection and Shengmai Injection on organ injury in endotoxemia rats].

As a dangerous disease with rapid progression, endotoxemia is easy to induce the damage to multiple organs. However, its specific and efficient treatment methods are still lacking at present. Both Qingkailing Injection(QKLI) and Shengmai Injection(SMI) have been proved effective in anti-inflammation, anti-endotoxin and organ protection. In this study, carrageenan and endotoxin were injected successively into rats to establish an endotoxemia model. Different doses of QKLI and SMI were administered to the endotoxemia rats by intraperitoneal injection separately or in combination. Then the count of white blood cells, the number of platelets, the content of cytokines, biochemical indexes, organ coefficient and pathological changes of main organs in the rats were detected. The results showed that the rats in the model group had obvious symptoms of endotoxemia, i.e., leucopenia, thrombocytopenia, increase in cytokines(IL-6 and TNF-α) and biochemical indexes of liver and kidney function as well as pathological damage to liver, kidney and lung. QKLI alone can alleviate the above symptoms of endotoxemia and the organ injury. SMI alone is less effective in improving disseminated intravascular coagulation(DIC) and cytokine secretion complicated with endotoxemia, but capable of reducing the inflammation degree of the lung, liver and kidney. The combination of QKLI and SMI remarkably increased the number of platelets in the peripheral blood, improved the liver and kidney function and reduced inflammatory factors, with lung, liver, kidney and other organ structures protected well. Moreover, the improvement effect of the combination of QKLI and SMI was stronger than those of the two injections alone at fixed doses, indicative of a synergistic effect.

[Overview of systematic reviews of Qingkailing Injection].

Qingkailing Injection is one of the most commonly used traditional Chinese medicine injections with significant clinical application for the treatment of multiple diseases. This study aims to analyze the systematic reviews( SRs) of Qingkailing Injection,in order to provide reference for the clinical application of Qingkailing Injection and the development of relevant clinical practice guidelines. We searched CNKI,CBM,Wanfang,VIP,Pub Med,Cochrane Library and EMbase to collect SRs from the time of database establishment to August 2020. The eligible SRs were included according to the inclusion and exclusion criteria. AMSTAR 2 was used to assess the methodological quality. The diseases,drugs in combinations and results were extracted and analyzed. A total of 24 SRs were selected,including 10 for the treatment of acute cerebrovascular diseases,9 for respiratory infections,2 for viral hepatitis,1 for chronic obstructive pulmonary disease,and two for the adverse effects of Qingkailing Injection. Only three entries of AMSTAR 2 item were fully reported by over 70%,and the rest were reported by less than 70%,with no report about item 2,3 and 10. Twenty-nine outcome indicators were correlated with the included SRs,of which three mostly frequent outcomes were effectiveness,adverse reaction,and neurological deficit scores,showing a good efficacy of Qingkailing Injection. The common severe adverse reaction was anaphylaxis,and mild adverse reactions were skin and mucous membrane reactions. The most frequently combined drug was antibiotics,mainly Penicillin and Penicillin+Pioneeromycin. The existing evidences showed that the methodological quality of SRs of Qingkailing Injection needed to be improved and Qingkailing Injection had an obvious efficacy. However,the selection of outcome indicators for clinical trials and SRs shall be standardized,and the reporting of basic information,such as drug combination,shall be strengthened to provide more powerful clinical services.

Network pharmacology assessment of Qingkailing injection treatment of cholestatic hepatitis.

OBJECTIVE: To investigate the targets and mechanisms of action of Qingkailing injection (，QKL) in the treatment of cholestatic hepatitis. METHODS: A network pharmacology method was implemented using drug and disease databases to target QKL and cholestasis hepatitis, respectively. The functional protein association network STRING database was used to construct a protein-protein interaction network using R language and the Bioconductor toolkit. The org.Hs.eg.db and clusterProfiler packages were used for gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, which explored biological functions and pathways of potential targets. Targets were then visualized using Cytoscape 3.6.0 software. RESULTS: We screened 121 compounds in QKL and identified 112 targets for the treatment of cholestatic hepatitis. QKL played a role in the treatment of cholestatic hepatitis through 305 biology process terms, 15 cellular component and 29 molecular function terms. The mechanism of QKL action was mainly related to tumor necrosis factor, mitogen-activated protein kinase, and PI3K-Akt signaling pathways. CONCLUSION: The treatment of cholestatic hepatitis by QKL involved multiple targets, biological functions, and signaling pathways that are closely associated with the disease.

Clinical Research of Qingkailing Injection in Adjuvant Treatment of Sepsis and Heat Syndrome in Children / 中国实验方剂学杂志

Objective:To evaluate the clinical efficacy of Qingkailing injection in the treatment of children with sepsis and heat syndrome， and investigate its anti-inflammatory， anticoagulant and protective effects. Method:Eighty patients were randomly divided into control group and observation group with forty cases in each group according to the number table. Both groups received comprehensive treatment measures such as fluid resuscitation， anti-infection， anti-inflammatory， anticoagulation， vasoactive drugs， and protection of vital organ functions. While patients in observation group additionally took Qingkailing injection， 5-10 mL each time， intravenous drip after dilution， 1 time/day. Treatment course was five days in both groups. Before and after treatment， the scores of quick sequential organ failure assessment （qSOFA）， pediatric critical illness score （PCIS） and acute physiology and chronic health evalution Ⅱ （APACHEⅡ） were graded； procalcitonin （PCT）， serum amyloid A protein （SAA） before and after treatment， heparin-binding protein （HBP）， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， high-sensitivity C-reactive protein （hs-CRP）， interleukin-6 （IL-6）， IL-10， N-terminal brain sodium Peptide precursor （NT-proBNP）， high-sensitivity cardiac troponin T （hs-cTnT） level， cardiac troponin I （cTnI）， creatine kinase isoenzyme （CK-MB）， <italic>D</italic>-dimer （<italic>D-</italic>D ）， fibrinogen （FIB） and antithrombin Ⅲ （AT-Ⅲ） levels were detected. Result:The APACHEⅡ and qSOFA scores in the observation group were lower than those in the control group （<italic>P</italic><0.05）， while the PCIS score was higher than that in the control group （<italic>P</italic><0.05）. The levels of PCT， SAA，HBP，TNF-<italic>α</italic>， hs-CRP and IL-6 in the observation group were lower than those in the control group （<italic>P</italic><0.01）， while the IL-10 level was higher than that in the control group （<italic>P</italic><0.01）. The levels of NT-proBNP， hs-cTnT， cTnI， CK-MB， <italic>D</italic>-D and FIB in the observation group were lower than those in the control group （<italic>P</italic><0.01）， while the AT-Ⅲ activity was higher than that in the control group （<italic>P</italic><0.01）. Conclusion:Qingkailing injection as the adjuvant therapy in children with sepsis and fever syndrome， can play the role of anti-inflammatory， anticoagulant， reducing infection and myocardial damage， thereby reducing the severity of the disease and improving the prognosis.

Synergistic Effect of Xiangdan Injection and Qingkailing Injection on Organ Injury Caused by Acute Inflammation and Thrombosis in Rats / 中国实验方剂学杂志

Objective:To investigate the synergistic effect of Xiangdan injection （XDI） and Qingkailing injection （QKLI） in the treatment of inflammation and thrombosis animal model based on changes of thrombus， inflammatory indexes， organ function， and pathological changes. Method:A total of 100 male SD rats were randomly divided into a normal control group， a model group， XDI groups （2.5， 5 mL·kg^-1）， QKLI groups （5， 10 mL·kg^-1）， and XDI + QKLI groups ［（2.5+5） mL·kg^-1，（2.5+10） mL·kg^-1，（5+5） mL·kg^-1，and （5+10） mL·kg^-1］ according to the body weight， with 10 rats in each group. Rats were treated correspondingly by intraperitoneal injection once a day for 4 days. The normal control group and the model group received normal saline. On the second day of administration， the model was induced in rats except those in the normal control group. Specifically， 25 mg·kg^-1carrageenan was injected intraperitoneally into the rats， followed by an injection of 50 μg·kg^-1 lipopolysaccharide （LPS） through the tail vein 16 hours later. Twenty-four hours after LPS injection， the rats were detected for liver index， kidney index， the number of platelets （PLT）， thrombus length， and biochemical indicators such as aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， alkaline phosphatase （ALP）， creatinine， and blood urea nitrogen （BUN）. Enzyme-linked immunosorbent assay （ELISA） was used to determine the content of inflammatory factors interleukin-6 （IL-6） and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of heart， liver， lung， and kidney， as well as the grading of organ injury. Result:Compared with the normal group， the model group showed decreased PLT， lengthened thrombus in the tail， increased liver index， elevated content of ALT， ALP， BUN， IL-6， and TNF-<italic>α</italic> （<italic>P</italic><0.05， <italic>P</italic><0.01）， and damaged liver， lung， and kidney tissues （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the conditions in the model group， XDI at 5 mL·kg^-1 reduced serum ALT and ALP in rats （<italic>P</italic><0.05， <italic>P</italic><0.01）， QKLI at 5 and 10 mL·kg^-1reduced serum levels ALT and ALP， and TNF-<italic>α </italic>content<italic> </italic>（<italic>P</italic><0.05， <italic>P</italic><0.01）. XDI at 5 mL·kg^-1 or QKLI at 10 mL·kg^-1 relieved the LPS-induced lung injury （<italic>P</italic><0.05）， the combination of XDI and QKLI decreased the levels of ALT， AST， ALP， and TNF-<italic>α， </italic>and the effect was predominant in the combination of XDI and QKLI at 5 and 10 mL·kg^-1（<italic>P</italic><0.05， <italic>P</italic><0.01）. Additionally， the length of the tail thrombus was significantly shortened （<italic>P</italic><0.05）， and the degree of lung injury was also reduced （<italic>P</italic><0.05）. The serum levels of ALT and BUN， TNF-<italic>α</italic> content， and liver index of rats were reduced after the combination of XDI and QKLI as compared with those in the single drug groups at the same dose （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:XDI or QKLI can improve or inhibit organ function， organ injury， and inflammatory response in the rat model of inflammation and thrombosis. The combination of the two drugs shows a synergistic effect in reducing the length of venous thrombus， improving liver and kidney function， inhibiting inflammatory factors， and protecting lung， liver， kidney， and other organs.

Study on synergistic effect of Qingkailing Injection and Shengmai Injection on organ injury in endotoxemia rats / 中国中药杂志

As a dangerous disease with rapid progression, endotoxemia is easy to induce the damage to multiple organs. However, its specific and efficient treatment methods are still lacking at present. Both Qingkailing Injection(QKLI) and Shengmai Injection(SMI) have been proved effective in anti-inflammation, anti-endotoxin and organ protection. In this study, carrageenan and endotoxin were injected successively into rats to establish an endotoxemia model. Different doses of QKLI and SMI were administered to the endotoxemia rats by intraperitoneal injection separately or in combination. Then the count of white blood cells, the number of platelets, the content of cytokines, biochemical indexes, organ coefficient and pathological changes of main organs in the rats were detected. The results showed that the rats in the model group had obvious symptoms of endotoxemia, i.e., leucopenia, thrombocytopenia, increase in cytokines(IL-6 and TNF-α) and biochemical indexes of liver and kidney function as well as pathological damage to liver, kidney and lung. QKLI alone can alleviate the above symptoms of endotoxemia and the organ injury. SMI alone is less effective in improving disseminated intravascular coagulation(DIC) and cytokine secretion complicated with endotoxemia, but capable of reducing the inflammation degree of the lung, liver and kidney. The combination of QKLI and SMI remarkably increased the number of platelets in the peripheral blood, improved the liver and kidney function and reduced inflammatory factors, with lung, liver, kidney and other organ structures protected well. Moreover, the improvement effect of the combination of QKLI and SMI was stronger than those of the two injections alone at fixed doses, indicative of a synergistic effect.

Overview of systematic reviews of Qingkailing Injection / 中国中药杂志

Qingkailing Injection is one of the most commonly used traditional Chinese medicine injections with significant clinical application for the treatment of multiple diseases. This study aims to analyze the systematic reviews( SRs) of Qingkailing Injection,in order to provide reference for the clinical application of Qingkailing Injection and the development of relevant clinical practice guidelines. We searched CNKI,CBM,Wanfang,VIP,Pub Med,Cochrane Library and EMbase to collect SRs from the time of database establishment to August 2020. The eligible SRs were included according to the inclusion and exclusion criteria. AMSTAR 2 was used to assess the methodological quality. The diseases,drugs in combinations and results were extracted and analyzed. A total of 24 SRs were selected,including 10 for the treatment of acute cerebrovascular diseases,9 for respiratory infections,2 for viral hepatitis,1 for chronic obstructive pulmonary disease,and two for the adverse effects of Qingkailing Injection. Only three entries of AMSTAR 2 item were fully reported by over 70%,and the rest were reported by less than 70%,with no report about item 2,3 and 10. Twenty-nine outcome indicators were correlated with the included SRs,of which three mostly frequent outcomes were effectiveness,adverse reaction,and neurological deficit scores,showing a good efficacy of Qingkailing Injection. The common severe adverse reaction was anaphylaxis,and mild adverse reactions were skin and mucous membrane reactions. The most frequently combined drug was antibiotics,mainly Penicillin and Penicillin+Pioneeromycin. The existing evidences showed that the methodological quality of SRs of Qingkailing Injection needed to be improved and Qingkailing Injection had an obvious efficacy. However,the selection of outcome indicators for clinical trials and SRs shall be standardized,and the reporting of basic information,such as drug combination,shall be strengthened to provide more powerful clinical services.

Application of traditional Chinese medicine Qingkailing to extend platelet storage in vitro: the metabolism and morphology study / 中国输血杂志

【Objective】 To study the metabolism and morphology changes of platelets in vitro using traditional Chinese medicine named Qingkailing injection as the additive solution, and to explore the viability of Qingkailing in the extension of platelet storage. 【Methods】 Apheresis platelets, adding 1% final concentration of Qingkailing injection, were taken as experiment groups, and sampled on 1, 3, 5, 8, 10 and 14 days(6 time points)of storage. Apheresis platelets without adding Qingkailing injection were taken as the control, and sampled on 1, 3 and 5 days(3 time points)of storage. The platelet count, morphology scores, biochemical parameters, pH and response rate of hypotonic shock during agitated storage(22 ℃) were tested. 【Results】 1) No significant change in platelet count was noticed in both experimental group(within 14 days) and the control(within 5 days)(P>0.05). The MPV and PDW of both groups increased at any sampling time within 5 days(P<0.05). 2) The morphology score of experimental groups and the control all decreased within the storage period(P<0.05), but its decrease of the control was greater than that of the experimental groups, especially on day 8(P<0.05). 3)Glucose, lactate dehydrogenase, lactate, Na+, and K+ values increased or decreased in varying degrees(P<0.05), while Cl- value stayed almost the same during 14 days(P>0.05). Glucose, lactate dehydrogenase, lactate and Na+ value changed significantly in the control within 5 days(P<0.05), while K+ and Cl- value did not(P>0.05). Within 5-day storage, the glucose consumption, lactate dehydrogenase and lactate generation in the control were significantly greater than those in experimental groups(P<0.05), but the added value of Na+, K+ and Cl- showed no significant difference(P>0.05). 4) pH value, relative to the baseline of day 1, decreased in both groups within 5 days, and its decreasing trend was significant in the control (P<0.05), but not in the experimental group(P>0.05). No significant difference was noticed in the response rate of hypotonic shock in experimental groups within 8 days, while significant decrease was noticed in the control within 5days(P<0.05). The response rate of hypotonic shock in experimental groups were significantly higher than that in the control on day 3 and day 5(P<0.05). 【Conclusion】 The comparison of apheresis platelets, stored in vitro, in terms of platelet count, morphology scores, biochemical parameters, pH and response rate of hypotonic shock showed that platelets, adding 1% final concentration of Qingkailing injection, could prolong the platelet storage to at least 10 days in vitro.

Exploring mechanism of Qingkailing Injection in treatment of coronavirus disease 2019 (COVID-19) based on network pharmacology and molecular docking / 中草药

Objective: To investigate the mechanism of Qingkailing Injection in the treatment of coronavirus disease 2019 (COVID-19). Methods: The active components and target proteins of Gardeniae Fructus, Isatidis Radix, Lonicerae Japonicae Flos, and other materials in Qingkailing Injection were obtained by means of literature search and TCMSP. Uniprot database was used to search the target genes corresponding to the active ingredients, and Cytoscape 3.7.2 was used to construct the drug-compound-target network. The enrichment analysis of KEGG pathway was carried out with the help of DAVID database to predict its mechanism. Core active components and potential targets of anti-COVID-19 drugs were verified by molecular docking. Results: The drug-compound- target network consisted of five drugs, 62 compounds and 70 targets. The KEGG pathway enrichment analysis included 41 signaling pathways (P < 0.05), which were mainly involved in cell apoptosis, Fc epsilon RI signaling pathway, TNF signaling pathway, etc. Molecular docking results showed that acacetin and syrigin had strong affinity with potential targets of anti-COVID-19 drugs. Conclusion: In this study, the effect of Qingkailing Injection has the characteristics of multiple components, multiple targets and multiple pathways. The active component, acacetin, can regulate the apoptosis pathway and TNF pathway by acting on CASP3, CASP8, FASLG, and other targets, so as to realize the potential therapeutic effect on COVID-19.

Qingkailing injection ameliorates cerebral ischemia-reperfusion injury and modulates the AMPK/NLRP3 Inflammasome Signalling pathway.

BACKGROUND: Cerebral ischemia is the second-leading cause of death and the main cause of permanent adult disabilities worldwide. Qingkailing (QKL) injection, a patented Chinese medicine approved by the China Food and Drug Administration, has been widely used in clinical practice to treat cerebral ischemia in China. The NOD-like receptor pyrin 3 (NLRP3) inflammasome is activated in cerebral ischemia and thus, is an effective therapeutic target. AMP-activated protein kinase (AMPK) is an important regulator inhibiting NLRP3 inflammasome activation. METHODS: We investigated the potential of QKL injection to provide neuroprotection after cerebral ischemia in a rat model of middle cerebral artery occlusion (MCAO). Adult male Sprague-Dawley rats (210-230 g) were randomly divided into three groups which consist of sham, MCAO and 3 ml/kg QKL. Rats in the QKL group received intraperitoneal injections of 3 ml/kg QKL, while rats in other groups were given saline in the same volumes. After 90 min ischemia and 24 h reperfusion, neurological function, laser speckle imaging, brain infarction, brain water content and brain blood barrier permeability were examined and cell apoptosis at prefrontal cortex were evaluated 24 h after MCAO, and western blot and real-time quantitative polymerase chain reaction was also researched, respectively. RESULTS: Intraperitoneal administration of QKL alleviated neurological deficiencies, cerebral infarction, blood-brain barrier permeability, brain oedema and brain cell apoptosis after MCAO induction. QKL decreased pro-inflammatory cytokines, TNF-α, IL-6 and IL-1ß, and increased anti-inflammatory cytokines, IL-4 and IL-10. Furthermore, QKL activated phosphorylated AMPK, decreased oxidative stress and decreased NLRP3 inflammasome activation. CONCLUSIONS: QKL relieved cerebral ischemia reperfusion injury and suppressed the inflammatory response by inhibiting AMPK-mediated activation of the NLRP3 inflammasome. These results suggest that QKL might have potential in treating brain inflammatory response and attenuating the cerebral ischemia-reperfusion injury.

[Analysis of Qingkailing Injection in treatment of combined medication features of 2 147 cases of upper respiratory tract infection].

Acute upper respiratory infection is one of the common infectious diseases,especially in the elderly people. Qingkailing Injection has the effect of clearing away heat and detoxifying. It can be used for external wind heat,upper respiratory tract infection,viral cold,etc. Based on 2 147 cases of upper respiratory tract infection patients using Qingkailing Injection in the medical electronic data warehouse of the information system of 16 large class-A hospitals constructed by the Institute of Basic Research in Clinical Medicine,China Academy of Chinese Medical Sciences,description analysis,Apriori calculation and recurrent network analysis were used to analyze the effect of Qingkailing Injection on upper respiratory tract infection. The clinical characteristics and the law of combined use of drugs can provide reference for the mining of clinical diagnosis and treatment characteristics and the construction of optimal and effective programs and paths. 2 147 patients with upper respiratory tract infection who used Qingkailing Injection were mostly middle-aged and elderly people over 45 years old,often combined with hypertension,ischemic cerebrovascular disease,coronary heart disease,diabetes,etc.; in treatment,it was more common to use Qingkailing Injection in combination with hormones,antipyretics and analgesics,antibiotics,nutritional support agents,antitussive,expectorant and antiasthmatic drugs,as well as traditional Chinese medicine Jiebiao agents and blood management agents. The combination of potassium chloride injection and cephalosporin injection is the most commonly used in combination with two kinds of Western medicine( 33. 2%); the combination of Shuanghuanglian and Ganmao Qingre Granules is the most commonly used in combination with two kinds of traditional Chinese medicine( 9. 13%); through the calculation of complex network group module,it is found that Qingkailing Injection is often associated with antibiotics,antipyretic and analgesic drugs,antitussive,expectorant and antiasthmatic drugs,antiviral drugs and anti infection drugs. Drugs,nutritional support agents,bronchodilators,immunomodulators and other chemical drugs are often used in combination with Chinese medicine,such as Jiebiao agents,expectorants,Qingre agents,resuscitation agents,Qufeng agents,tonics,Liqi agents,which may be related to both the treatment of upper respiratory tract infection and the basic diseases of middle-aged and elderly patients,with the theoretical characteristics of " combination of disease and syndrome,corresponding formula and syndrome". Based on the real world big data complex network group module mining results provide clues for the clinical optimization scheme and path construction,and provide signals and ideas for further causal research.

Effect of Qingkailing on Expression of Toll-like Receptor 4, gp91phox and Zonula Occludens-1 in Cerebrovascular Endothelial Cells Induced by Hypoxia Activating Microglias / 中国康复理论与实践

Objetive To investigate the effect of Qingkailing injection on the expression of Toll-like receptor 4 (TLR4), gp91phox and zonula occludens-1 (ZO-1) in cerebrovascular endothelial cells induced by hypoxia activation of microglias. Methods:BV2 microglia cells were divided into six groups. They were cultured in serum-free DMEM, while the Qingkailing groups of low, middle and high dosages were cultured with 0.0625%, 0.125% and 0.25% Qingkailing injection, respectively, and minocycline group with minocycline of 200 nmol/L. The groups other than control group underwent hypoxia for 24 hours and reoxygenation for 24 hours. Then, the medium of microglia was put into the medium of Balb/c endothelial cells for 24 hours. The cell viability of endothelial cells was measured with CCK-8, the concentration of nitric oxide (NO) was detected with colorimetry, and the experission of TLR4, gp91phox and ZO-1 was detected with Western blotting. Results:Compared with the control group, the cell viability and the expression of ZO-1 decreased in the model group (P < 0.01), while the concentration of NO and the expression of TLR4 and gp91phox increased (P < 0.05). Compared with the model group, the cell viability and the expression of ZO-1 increased in the Qingkailing groups and the minocycline group (P < 0.05), while the concentration of NO and the expression of TLR4 and gp91phox decreased (P < 0.05). Conclusion:Qingkailing injection may enhance the survival and function of cerebrovascular endothelial cells by inhibiting the hypoxia activation of microglias, reducing the expression of TLR4 and gp91phox, and increasing the expression of ZO-1.

Analysis of Qingkailing Injection in treatment of combined medication features of 2 147 cases of upper respiratory tract infection / 中国中药杂志

Acute upper respiratory infection is one of the common infectious diseases,especially in the elderly people. Qingkailing Injection has the effect of clearing away heat and detoxifying. It can be used for external wind heat,upper respiratory tract infection,viral cold,etc. Based on 2 147 cases of upper respiratory tract infection patients using Qingkailing Injection in the medical electronic data warehouse of the information system of 16 large class-A hospitals constructed by the Institute of Basic Research in Clinical Medicine,China Academy of Chinese Medical Sciences,description analysis,Apriori calculation and recurrent network analysis were used to analyze the effect of Qingkailing Injection on upper respiratory tract infection. The clinical characteristics and the law of combined use of drugs can provide reference for the mining of clinical diagnosis and treatment characteristics and the construction of optimal and effective programs and paths. 2 147 patients with upper respiratory tract infection who used Qingkailing Injection were mostly middle-aged and elderly people over 45 years old,often combined with hypertension,ischemic cerebrovascular disease,coronary heart disease,diabetes,etc.; in treatment,it was more common to use Qingkailing Injection in combination with hormones,antipyretics and analgesics,antibiotics,nutritional support agents,antitussive,expectorant and antiasthmatic drugs,as well as traditional Chinese medicine Jiebiao agents and blood management agents. The combination of potassium chloride injection and cephalosporin injection is the most commonly used in combination with two kinds of Western medicine( 33. 2%); the combination of Shuanghuanglian and Ganmao Qingre Granules is the most commonly used in combination with two kinds of traditional Chinese medicine( 9. 13%); through the calculation of complex network group module,it is found that Qingkailing Injection is often associated with antibiotics,antipyretic and analgesic drugs,antitussive,expectorant and antiasthmatic drugs,antiviral drugs and anti infection drugs. Drugs,nutritional support agents,bronchodilators,immunomodulators and other chemical drugs are often used in combination with Chinese medicine,such as Jiebiao agents,expectorants,Qingre agents,resuscitation agents,Qufeng agents,tonics,Liqi agents,which may be related to both the treatment of upper respiratory tract infection and the basic diseases of middle-aged and elderly patients,with the theoretical characteristics of " combination of disease and syndrome,corresponding formula and syndrome". Based on the real world big data complex network group module mining results provide clues for the clinical optimization scheme and path construction,and provide signals and ideas for further causal research.

Deciphering biochemical basis of Qingkailing injection-induced anaphylaxis in a rat model by time-dependent metabolomic profiling based on metabolite polarity-oriented analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Qingkailing injection (QKLI) is prepared from eight traditional Chinese medicinal materials or their extracts, which is widely used in clinical practice to treat the upper respiratory inflammation, pneumonia, high fever and viral encephalitis, nonetheless, suffering from serious anaphylaxis. AIM OF STUDY: This study aims to develop an integrative metabolomics approach for deciphering the biochemical basis of QKLI induced anaphylaxis (QKLI-IA). MATERIALS AND METHODS: The accuracy of animal modeling, the coverage of detected metabolites and the timeliness of pathological reaction are three key factors for revealing the biochemical basis of disease with untargeted metabolomics. In this study, firstly, the allergic rats (responders) were first screened by passive cutaneous anaphylaxis experiment and then were utilized for modeling. To cover a wider range of metabolites, a large-scale untargeted metabolomics based on metabolites polarity-oriented analysis was performed using ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Then, to evaluate the timeliness of QKLI-IA, a time-dependent metabolomic profiling including the early, mid and late anaphylaxis stages of QKLI-IA, was performed. RESULTS: Corresponding to early, mid and late anaphylaxis stages of QKLI-IA, 14, 9 and 4 potential biomarkers were identified, respectively. Metabolism pathway analysis revealed that QKLI-IA resulted in dynamic changes in serum amino acid, fatty acid, glycerolipid, and phospholipid metabolisms. Twenty-four metabolites were found with identical fluctuating trends across the three stages of QKLI-IA. The results indicate that the pathogenesis of QKLI-IA is closely related to arachidonic acid metabolism. CONCLUSION: This research provides a methodology reference for revealing the biochemical basis of disease using metabolomic profiling and offers a new insight to understand the pathogenesis of QKLI-IA.

[Pseudoallergic reaction characteristics of Qingkailing injection and preliminary screening of allergic substances].

This study aimed to explore the characteristics and the influencing factors of Qingkailing injection (QKLI) pseudoallergic reaction, and screen out the possible pseudoallergenic substances. The results showed that ICR and Kunming mice had stronger pseudoallergic reactions than BALB/c and C57 mice after being injected with the same dose of QKLI. The pseudoallergic reaction induced by QKLI that was prepared with 0.9% saline was stronger than that prepared with 5% glucose. When the dose was twice of the clinical dose, some batches of QKLI could cause significant or suspected pseudoallergic reactions; when the dose dropped to clinically equal times, all of the batches did not induce pseudoallergic reactions in mice. Different batches of QKLI induced different pseudoallergic reactions in mice. Therefore, QKLI's pseudoallergic reactions might have a certain relationship with different body constitutions. Different solvents might affect the safety of QKLI. QKIL-induced pseudoallergic reactions had the different characteristics between batches, and the dosage should be strictly controlled in clinical use. After the comparison of pseudoallergic reactions induced by different components and different intermediates of QKLI in mice, it was preliminary believed that pseudoallergenic substances might exist in intermediate Isatidis Radix extracts and Gardenia extracts, but specific pseudoallergens shall be furthered studied in subsequent experiences.

Qingkailing Injection () for Treatment of Children Pneumonia Induced by Respiratory Syncytial Virus: A Meta-Analysis of Randomized Controlled Trials.

OBJECTIVE: To evaluate the efficacy and safety of Qingkailing Injection (, QKL) for treatment of children pneumonia caused by respiratory syncytial virus (RSV). METHODS: Randomized clinical trials (RCTs) comparing QKL with ribavirin injection in the treatment of children pneumonia induced by RSV were searched in PubMed, Science Direct, Cochrane Library, Chinese VIP database, CNKI and Wanfang databases from their inception to March 2014. Meta-analyses were performed using RevMan 5.2 software. The methodological quality of the selected RCTs was evaluated by the Modified Jadad Score. The primary outcome measures were effective rate and the secondary outcomes were relief time of fever and cough. RESULTS: Seven RCTs with 992 cases published from 2008 to 2013 were identified. The meta-analysis results indicated that QKL was more effective in cure rate [risk ratios (RR)=1.32, 95% CI (1.17, 1.50), P<0.01], total effective rate [RR=1.07, 95% CI (1.02, 1.13), P=0.009] and less fever clearance time [mean difference=-0.73, 95% CI (-1.22,-0.23), P=0.004], compared with ribavirin injection in the treatment of RSV-induced children pneumonia. No dead case was reported in all trials. There were 3 trials mentioned adverse events, 2 reported no obvious adverse event occurred while 1 reported adverse events described as skin hypersensitivity, elevation of ALT, a mild abnormal of hepatic and renal function in both QKL and ribavirin group. CONCLUSIONS: QKL was an effective and relatively safe option for the treatment of RSV-induced children pneumonia. These therapeutic effects were promising but need to be interpreted with caution due to variations in the treatment and methodological weakness in the studies.

Multiple perspectives of qingkailing injection-fraction-single compound in revealing the hepatotoxicity of baicalin and hyodeoxycholic acid.

ETHNOPHARMACOLOGICAL RELEVANCE: The complexity of ingredients in traditional Chinese medical formulas and the limited consideration of toxicological responses are fundamental issues that hamper prognostic information of drug quality control. MATERIALS AND METHODS: A multidisciplinary approach for quality control of Qingkailing injection (QKL) regarding drug induced liver toxicity was described for the first time. High content image analysis (HCA) was combined with reverse-phase chromatographic separation and high-resolution MS detection technologies to provide the dynamic responses of drug induced HepG2 cell injury. Firstly, a simple and rapid method for simultaneous qualification and quantification of 21 major constituents in QKL was established and validated using ultra-high performance liquid chromatography-hybrid quadrupole-Orbitrap mass spectrometer (UHPLC-Q-Orbitrap), which were operated in full MS/dd-MS2 mode and thus simultaneously acquired quantitative high resolution (HR) full scan data and confirmatory HR MS2 data. Secondly, repeated semi-preparation HPLC was applied to obtain four fractions (F1-F4) for HCS analysis. Finally, potential hepatotoxicity was determined by five hepatotoxicity biomarkers, including cell loss, DNA condense, glutathione (GSH) depletion, reactive oxygen species (ROS) formation, and mitochondria membrane potential (MMP) depolarization. RESULTS: The detection in polarity switching mode empowered the coverage of comprehensive constituents with different chemical properties. Satisfactory linearity precisions, repeatability, stability, and recovery were achieved. QKL injection significantly induced HepG2 cell injury above the concentration of 1.25% (v/v). Meanwhile, flavone glycosides (F3) and stinasterols (F4) fractions exhibited hepatotoxicity above 75µg/mL and 50µg/mL, respectively. Still further, baicalin originated from F3 significantly caused cell loss and glutathione (GSH) depletion. In parallel, hyodeoxycholic acid from F4 induced cell loss, nucleus condense, and GSH reduction as well. CONCLUSIONS: Our work provides multiple perspectives based on injection-fractions-single compound format to improve QKL pharmacovigilance through revealing the potential hepatotoxic material basis. Additionally, our study provides an integrating paradigm for the comprehensive and systematic quality control of traditional Chinese medical formulas.

Pseudoallergic reaction characteristics of Qingkailing injection and preliminary screening of allergic substances / 中国中药杂志

This study aimed to explore the characteristics and the influencing factors of Qingkailing injection (QKLI) pseudoallergic reaction, and screen out the possible pseudoallergenic substances. The results showed that ICR and Kunming mice had stronger pseudoallergic reactions than BALB/c and C57 mice after being injected with the same dose of QKLI. The pseudoallergic reaction induced by QKLI that was prepared with 0.9% saline was stronger than that prepared with 5% glucose. When the dose was twice of the clinical dose, some batches of QKLI could cause significant or suspected pseudoallergic reactions; when the dose dropped to clinically equal times, all of the batches did not induce pseudoallergic reactions in mice. Different batches of QKLI induced different pseudoallergic reactions in mice. Therefore, QKLI's pseudoallergic reactions might have a certain relationship with different body constitutions. Different solvents might affect the safety of QKLI. QKIL-induced pseudoallergic reactions had the different characteristics between batches, and the dosage should be strictly controlled in clinical use. After the comparison of pseudoallergic reactions induced by different components and different intermediates of QKLI in mice, it was preliminary believed that pseudoallergenic substances might exist in intermediate Isatidis Radix extracts and Gardenia extracts, but specific pseudoallergens shall be furthered studied in subsequent experiences.

Qingkailing Injection () for Treatment of Children Pneumonia Induced by Respiratory Syncytial Virus: A Meta-Analysis of Randomized Controlled Trials / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of Qingkailing Injection (, QKL) for treatment of children pneumonia caused by respiratory syncytial virus (RSV).</p><p><b>METHODS</b>Randomized clinical trials (RCTs) comparing QKL with ribavirin injection in the treatment of children pneumonia induced by RSV were searched in PubMed, Science Direct, Cochrane Library, Chinese VIP database, CNKI and Wanfang databases from their inception to March 2014. Meta-analyses were performed using RevMan 5.2 software. The methodological quality of the selected RCTs was evaluated by the Modified Jadad Score. The primary outcome measures were effective rate and the secondary outcomes were relief time of fever and cough.</p><p><b>RESULTS</b>Seven RCTs with 992 cases published from 2008 to 2013 were identified. The meta-analysis results indicated that QKL was more effective in cure rate [risk ratios (RR)=1.32, 95% CI (1.17, 1.50), P<0.01], total effective rate [RR=1.07, 95% CI (1.02, 1.13), P=0.009] and less fever clearance time [mean difference=-0.73, 95% CI (-1.22,-0.23), P=0.004], compared with ribavirin injection in the treatment of RSV-induced children pneumonia. No dead case was reported in all trials. There were 3 trials mentioned adverse events, 2 reported no obvious adverse event occurred while 1 reported adverse events described as skin hypersensitivity, elevation of ALT, a mild abnormal of hepatic and renal function in both QKL and ribavirin group.</p><p><b>CONCLUSIONS</b>QKL was an effective and relatively safe option for the treatment of RSV-induced children pneumonia. These therapeutic effects were promising but need to be interpreted with caution due to variations in the treatment and methodological weakness in the studies.</p>