Single-cell transcriptome analysis reveals the regulatory effects of artesunate on splenic immune cells in polymicrobial sepsis

Sepsis is characterized by a severe and life-threatening host immune response to polymicrobial infection accompanied by organ dysfunction.Studies on the therapeutic effect and mechanism of immunomodulatory drugs on the sepsis-induced hyperinflammatory or immunosuppression states of various immune cells remain limited.This study aimed to investigate the protective effects and underlying mechanism of artesunate(ART)on the splenic microenvironment of cecal ligation and puncture-induced sepsis model mice using single-cell RNA sequencing(scRNA-seq)and experimental validations.The scRNA-seq analysis revealed that ART inhibited the activation of pro-inflammatory macrophages recruited during sepsis.ART could restore neutrophils’chemotaxis and immune function in the septic spleen.It inhibited the activation of T regulatory cells but promoted the cytotoxic function of natural killer cells during sepsis.ART also promoted the differentiation and activity of splenic B cells in mice with sepsis.These results indicated that ART could alleviate the inflammatory and/or immunosuppressive states of various immune cells involved in sepsis to balance the immune homeostasis within the host.Overall,this study provided a comprehensive investigation of the regulatory effect of ART on the splenic microenvironment in sepsis,thus contributing to the application of ART as adjunctive therapy for the clinical treatment of sepsis.

Tumor microenvironment-responsive artesunate loaded Z-scheme heterostructures for synergistic photo-chemodynamic therapy of hypoxic tumor

Tumor microenvironment(TME)with the particular features of severe hypoxia,insufficient endogenous H2O2,and overexpression of glutathione(GSH)markedly reduced the antitumor efficacy of monotherapy.Herein,a TME-responsive multifunctional nanoplatform(Bi2S3@Bi@PDA-HA/Art NRs)was presented for synergistic photothermal therapy(PTT),chemodynamic therapy(CDT),and photodynamic therapy(PDT)to achieve better therapeutic outcomes.The Z-scheme heterostructured bismuth sulfide@bismuth nanorods(Bi2S3@Bi NRs)guaranteed excellent photothermal performance of the nanoplatform.Moreover,its ability to produce O2 and reactive oxygen species(ROS)synchronously could relieve tumor hypoxia and improve PDT outcomes.The densely coated polydopamine/ammonium bicarbonate(PDA/ABC)and hyaluronic acid(HA)layers on the surface of the nanoplatform enhanced the cancer-targeting capacity and induced the acidic TME-triggered in situ“bomb-like”release of Art.The CDT treatment was achieved by activating the released Art through intracellular Fe2+ions in an H2O2-independent manner.Furthermore,decreasing the glutathione peroxidase 4(GPX4)levels by Art could also increase the PDT efficiency of Bi2S3@Bi NRs.Owing to the synergistic effect,this nanoplatform displayed improved antitumor efficacy with minimal toxicity both in vitro and in vivo.Our design sheds light on the application of phototherapy combined with the traditional Chinese medicine monomer-artesunate in treating the hypoxic tumor.

The Combination of Artesunate and Paclitaxel in 1:1 Ratio Induces Apoptosis and Morphology Change on Human Prostate Cancer Cell Lines

The combination of Artesunate (ART) and Paclitaxel (PTX) in two human prostate cancer (PCa) cell lines (PC-3 and LNCaP) was evaluated to investigate the effects on proliferation, apoptosis and morphological changes. The half maximal inhibitory concentration (IC50) values that were observed by ART and PTX on both LNCaP and PC-3 cell lines at 72-and 120-hour exposure were used to assess these effects. Early and late apoptosis was detected in Annexin V-FITC/PI assay revealed a shift in population of cells towards early and mid-apoptosis with ART + PTX than with ART and PTX individually. More effects were observed on LNCaP cell lines at both 72-hour and 120-hour exposure. The results for the Caspase 3/7 activity assay showed shift of viable population in all induced samples compared to control. Morphological changes occurred in both cell lines;this was validated in qualitative assessment when examined under the inverted microscope. These findings indicated that ART + PTX suppressed PCa cell proliferation in a dose- and time-dependent manner.

Research progress of artesunate in diabetes and its complications

Diabetes is a metabolic disease characterized by abnormally elevated blood glucose levels.Persistent hyperglycemia leads to diabetic nephropathy,diabetic retinopathy,diabetes with periodontal disease and other diabetic complications.These diseases have become the main causes of disability and death in diabetic patients.Artesunate is well known as an antimalarial drug for controlling malaria symptoms.Current studies have shown that artesunate improves diabetes and its complications by protecting islet cells,improving glucose and lipid metabolism,anti-inflammatory and immune regulation.Based on the research status in recent years,this paper focuses on the mechanism of artesunate in diabetes and its complications,to provide a theoretical basis for future diabetes research.

Artesunate对克罗恩病样结肠炎的保护性作用及机制

目的探讨青蒿琥酯(Artesunate)对TNBS结肠炎小鼠的治疗作用及机制。方法采用6～8周龄Balb/c小鼠,建立TNBS模型后将16只小鼠随机分为Artesunate组及Model组两组,每组8只。Artesunate组小鼠每日腹腔注射Artesunate(150 mg/kg/d),Model组小鼠每日给予腹腔注射等量的生理盐水。干预4周后处死,采用病理组织学、分子免疫学等技术评估肠炎、肠屏障功能及JAK2/STAT3信号改变。结果干预第3～4周,Artesunate组小鼠DAI评分显著低于Model组小鼠(P<0.05),而体重显著高于Model组小鼠(P<0.05)。治疗后,Artesunate组小鼠肠道炎症组织学评分显著低于Model组小鼠(P <0.05)。同时,Artesunate组小鼠肠黏膜IL-1β及TNF-α水平显著低于Model组小鼠(P<0.05),而IL-10水平显著高于Model组(P<0.05)。Artesunate组小鼠血清FITC水平和肠系膜淋巴结与肝脏的细菌移位比例均显著低于Model组小鼠(P<0.05)。Artesunate组小鼠肠黏膜p-JAK2及p-STAT3水平均显著低于Model组小鼠(P<0.05)。结论 Artesunate治疗可通过抑制JAK2/STAT3信号,发挥保护肠黏膜屏障、抑制肠黏膜炎症反应等抗CD样肠炎的作用。

Effect of artesunate supplementation on bacterial translocation and dysbiosis of gut microbiota in rats with liver cirrhosis

AIM: To evaluate the effect of artesunate(AS) supplementation on bacterial translocation(BT) and gut microbiota in a rat model of liver cirrhosis. METHODS: Fifty-four male Sprague-Dawley rats were randomly divided into a normal control group(N), a liver cirrhosis group(M) and a liver cirrhosis group intervened with AS(MA). Each group was sampled at 4, 6 and 8 wk. Liver cirrhosis was induced by injection of carbon tetrachloride(CCl4), intragastric administration of 10% ethanol, and feeding a high fat diet. Rats in the MA group were intragastrically administered with AS(25 mg/kg body weight, once daily). Injuries of the liver and intestinal mucosa were assessed by hematoxylineosin or Masson's trichrome staining. Liver index was calculated as a ratio of the organ weight(g) to body weight(g). The gut microbiota was examined by automated ribosomal intergenic-spacer analysis of fecal DNA. BT was assessed by standard microbiological techniques in the blood, mesenteric lymph nodes(MLNs), liver, spleen, and kidney. RESULTS: Compared to group N, the body weight was reduced significantly in groups M and MA due to the development of liver cirrhosis over the period of 8 wk. The body weight was higher in group MA than in group M. The liver indices were significantly elevated at 4, 6 and 8 wk in groups M and MA compared to group N. AS supplementation partially decreased the liver indices in group MA. Marked histopathologic changes in the liver and small intestinal mucosa in group M were observed, which were alleviated in group MA. Levels of pro-inflammatory interleukin-6 and tumor necrosis factor-α were significantly elevated at 8 wk in ileal homogenates in group M compared to group N, which were decreased after AS supplementation in group MA. The dysbiosis of gut microbiota indicated by the mean diversity(Shannon index) and mean similarity(Sorenson index) was severe as the liver cirrhosis developed, and AS supplementation had an apparent intervention effect on the dysbiosis of gut microbiota at 4 wk. The occurrence of BT was increased in the liver of group M compared to that of group N. AS supplementation reduced BT in group MA at 8 wk. BT also occurred in the MLNs, spleen, and kidney, which was reduced by AS supplementation. BT was not detected in the blood in any group.CONCLUSION: Dysbiosis of gut microbiota, injury of intestinal mucosal barrier and BT occurred as liver cirrhosis progressed, which might enhance inflammation and aggravate liver injury. AS may have other nonantimalarial effects that modulate gut microbiota,inhibit BT and alleviate inflammation, resulting in a reduction in CCl4, alcohol and high fat-caused damages to the liver and intestine.

Antitumor effects of artesunate on human breast carcinoma MCF-7 cells and IGF-IR expression in nude mice xenografts

Purpose： The objective of this study was to investigate the anti-tumor effects and analyze the mechanism of artesunate （ART） action on breast cancer in vivo using tumor transplanted nude mice. Methods： The human breast tumor cell line MCF-7 was transplanted into nude mice, and the animals were treated with various doses of ART alone or in combination with cyclophosphamide （CTX） or normal saline （NS）. The tumor inhibitory effects were observed and compared, and the ultrastructural morphology of the transplanted tumor cells was observed by electron microscopy. The apoptosis rates and cell cycle status were detected by flow cytometry （FCM）. The expression of apoptosis-related proteins p53, Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry and IGF-IR was detected by western blot. The expression correlation for these proteins was also analyzed. Results： The tumor inhibition rates in the low dose ART group, high dose ART group, CTX group and combined drug therapy group were （24.39±10.20）%, （40.24±7.02）%, （57.01±5.84）% and （68.29±5.1）%, respectively. The cell cycle was arrested in phase G0/Gt after treatment with ART. The expression of Bcl-2 was significantly reduced, and the expression levels of Bax and Caspase-3 were significantly increased in the ART group compared to the negative control saline group. There was no significant difference detected in p53 expression. The Bcl-2 level was negatively related to Bax and Caspase-3. The western blotting results showed IGF-IR downregulation. Conclusions： ART inhibits the growth of MCF-7 breast tumor cell xenografts in nude mice. The anti-tumor mechanism of ART for human breast carcinoma in nude mice might be correlated with the alteration of apoptosis related protein expression, which may further induce apoptosis and inhibit cell proliferation.

Artesunate Effect on Schistosome Thioredoxin Glutathione Reductase and Cytochrome c Peroxidase as New Molecular Targets in Schistosoma mansoni-infected Mice

Objective To investigate the possible effect of artesunate （ART） on schistosome thioredoxin glutathione reductase （TGR） and cytochrome c peroxidase （CcP） in Schistosoma mansoni-infected mice. Methods A total of 200 laboratory bred male Swiss albino mice were divided into 4 groups （50 mice in each group）. Group I： infected untreated group （Control group） received a vehicle of 1% sodium carbonyl methylcellulose （CMC-Na）; Group II： infected then treated with artesunate; Group III infected then treated with praziquantel, and group IV： infected then treated with artesunate then praziquantel. Adult S. mansoni worms were collected by Animal Perfusion Method, tissue egg counted, TGR, and CcP mRNA Expression were estimated of in $. mansoni adult worms by semi-quantitative rt-PCR. Results Semi-quantitative rt-PCR values revealed that treatment with artesunate caused significant decrease in expression of schistosome TGR and CcP in comparison to the untreated group. In contrast, the treatment with praziquantel did not cause significant change in expression of these genes. The results showed more reduction in total worm and female worm count in combined ART-PZQ treated group than in monotherapy treated groups by either ART or PZO, Moreover, complete disappearance （100%） of tissue eggs was recorded in ART-PZQ treated group with a respective reduction rate of 95.9% and 68.4% in ART- and PZQ-treated groups. Conclusion The current study elucidated for the first time that anti-schistosomal mechanisms of artesunate is mediated via reduction in expression of schistosome TGR and CcP. Linking these findings, addition of artesunate to praziquantel could achieve complete cure outcome in treatment of schistosomiasis.

Artesunate inhibits proliferation and migration of RPE cells and TGF-β2 mediated epithelial mesenchymal transition by suppressing PI3K/AKT pathway

AIM:To study the braking effectiveness of artesunate on transforming growth factor(TGF)-β2 mediated epithelial-mesenchymal transition(EMT)in retinal pigment epithelium(RPE)in vitro.METHODS:The fostered ARPE-19 cells were processed with artesunate alone or combined with the TGF-β2.The CCK-8 examination was utilized to test the cell propagation.Cell migration was detected by scratch as well as the Transwell examination.The EMT characters and activation of PI3K/Akt signal channel were estimated by Western blotting and immunofluorescence.The Western blotting was utilized in order to confirm the vitreous of controls as well as patients with proliferative vitreoretinopathy(PVR)were collected and the levels of PI3K,phospho-PI3K,Akt.RESULTS:Disposal of ARPE-19 cells with artesunate(50-150μmol/L)obviously suppressed their propagation and immigration,which dependent on the concentration and time.Artesunate suppressed the EMT which was induced by TGF-β2 in ARPE-19 cells through sustaining the expression of vimentin andα-SMA through the suppression of PI3K,phospho-PI3K,phospho-Akt and Akt.Levels of PI3K,phospho-PI3K,AKT and phospho-Akt was increased in the vitreous in PVR(P<0.05).CONCLUSION:Such findings indicate that PI3K/Akt signal channel is highly activated in vitreous of PVR.Artesuante is an operative depressor of the propagation,immigration and TGF-β2-mediated EMT of ARPE-19 cells by reduced the expression of PI3K/Akt channel.

Anti-malaria drug artesunate protects bronchial epithelium from DNA damage induced by asthma

OBJECTIVE To investigate the genome protective effects of anti-malaria drug,artesunate in an experimental allergic asthma model.METHODS Mice were sensitized on day 0 and 7 and challenged on day 14 with 100μg house dust mite(HDM)via intratracheal administration.Artesunate(30mg·kg-1)was administered intra-peritoneally on day 6,7,8,13,14 and 15.Samples were collected on day 1,3 and 5 post last HDM-challenge for analysis of air way inflammation and DNA damage.Lung sections were immunofluorescence(IF)-stained for DNA double strand breaks(DSBs)markers,γH2AX and 53BP1.Levels of DNA repair proteins Ku70 and Rad51,which are involved in non-homologous end joining(NHEJ)and homologous recombination(HR)DNA DSB repair pathways respectively,were measured.To quantify cell death in asthmatic lung,TUNEL staining was performed.Comet assay,a single cell gel electrophoresis was employed to detect DNA damage induced by HDM in BEAS-2Bhuman bronchial epithelial cell line,in vitro.RESULTS Artesunate treatment significantly reduces immune cells infiltration in BAL fluid of asthmatic mice,collected on day 3 and 5 post-challenge.Importantly,artesuante is able to protect bronchial epithelium from DNA DSBs induced by asthma,as detected by the reduced level of γH2AX and 53BP1 foci formation in the nucleus.This genome protective effect is evident even on day 1 post-challenge,when immune cells infiltration remained high.This indicates that artesunate confers protection on bronchial epithelium in the presence of inflammation.Additionally,artesunate is also able to reduce cell death in asthmatic lung revealed by TUNEL assay and cleaved caspase 3 level.Interestingly,the levels of DNA repair proteins in artesuante-treated asthmatic mice are unchanged as compared to HDM-only mice,suggesting that artesunate treatment does not augment the level of DNA repair proteins.When human bronchial epithelial BEAS-2 Bcells were exposed to HDMin vitro,we observed an increase in the levels of DNA damage.Artesunate(60μmol·L-1)co-incubated with HDM is not able to prevent direct DNA damage induced by the allergen.Together,these studies suggest that the genome protective effect of artesunate in vivo may be attributed to physiological effects(such as its anti-inflammatory effects)rather than serving to directly prevent DNA damage.CONCULSION This study highlights a novel role for artesunate in protecting bronchial epithelial cells from asthma-induced DNA damage.

Benefits of Artesunate versus Quinine in the Treatment of Children with Severe Malaria at the National University Teaching Hospital of Cotonou

Introduction: Severe malaria is one of the leading causes of death in Sub-Saharan African countries, and artesunate is recommended as a first-line treatment by the Word Heath Organization (WHO.). Objective: Identify the advantages of artesunate compared with quinine in the treatment of severe malaria in children. Methods and patients: This study was a cross-sectional, descriptive and analytical study focused on children hospitalized for severe malaria in the CNHU who were treated with quinine or artesunate. Findings: The hospital-based frequency rate of severe malaria in pediatric patients was estimated to be 28.3% (n = 848). One hundred five children were treated with artesunate, and 743 were treated with quinine. The mean age of the children was 47 months old. The primary signs of severity were anemia (n = 776), neurological manifestations (n = 309) and hemolysis (n = 137). The average duration of treatment was 1.95 days for artesunate versus 2.45 days for quinine, and the difference was statistically significant (p = 0.001). The average length of stay (ALOS) in the hospital was 5 days for the artesunate group versus 5.75 days for the quinine group, and the difference was statistically significant (p < 0.001). Six of the children who received artesunate died, whereas 24 children who treated with quinine died. The total average cost of healthcare was 50,600 FCFA (77 euros) per child treated with artesunate versus 57,100 FCFA (87 euros) per child treated with quinine. Conclusion: The treatment of severe malaria with artesunate is superior to quinine-based treatment.

Antimalarial activity of Ageratum conyzoides in combination with chloroquine and artesunate

Objective:To determine the suppressive and curative activity of aqueous leaf extract of Ageratum conyzoides(A.conyzoides) in combination with chloroquine and artesunate, respectively against Plasmodium berghei infection in mice.Methods:Using malaria(Plasmodium berghei) infected albino mice of both sexes,aqueous extracts of A.conyzoides in combination with chloroquine and artesunate were tested for antimalarial activity,respectively.Four-day suppressive test and Rane’s curative test were carried out.Results:Suppressive tests showed significant dose dependent reduction in parasitemia level produced by the extract-chloroquine and extract-artesunate combinations.Suppressive activities of both extract-drug combinations were greater than the individual drugs alone.Extract-chloroquine(100:5) produced the highest suppressive effect(98%suppression).Curative tests showed absolute survival in two extract-drug combinations.Two extract-drug combinations produced higher curative effects than the individual drugs alone.The highest dose combinations of extract-chloroquine(100:5) and extract-artesunate(100:5) produced absolute parasitemia clearance(cure) in the infected mice. Conclusions:The study indicated that aqueous extract of A.conyzoides had the ability to potentiate the antimalarial activity of chloroquine and artesunate against induced plasmodiasis in mice.It contributes a lot in the malaria endemic and poverty stricken tropics.

Effects of albendazole,artesunate,praziquantel and miltefosine,on Opisthorchis viverrini cercariae and mature metacercariae

Objective:To explore larvicidal effects of anthelmintic drugs on Opisthorchis viverrini(O.viverrini) for alternative approach to interrupting its cycle for developing a field-based control program.Methods:The larvicidal activities of albendazole(A1),artesunate(Ar),praziquantel(Pzq) and miltefosine(Mf) on O.viverrini cercariae and mature metacercariae were investigated.Lethal concentrations(LC_(50) and LC_(95)) of these drugs were determined.Mature metacercariae previously exposed to various concentrations of the drugs were administered to hamsters.Worms were harvested 30 d post infection and worm recovery rates calculated.Al,Ar,Pzq and Mf produced morphological degeneration and induced shedding tails of cercariae after 24 h exposure.Results:The LC_(50) and LC_(95) of Al,Ar,Pzq and Mf on cercariae were 0.720 and 1.139,0.350 and 0.861,0.017 and 0.693,and 0.530 and 1.134 ppm,respectively.LC_(50) and LC_(95) of Ar on mature metacercariae were 303.643 and 446.237 ppm and of Mf were 289.711 and 631.781 ppm,respectively but no lethal effect in Pzq-and Al-treated groups(up to 1 ppt).No worms were found in hamsters administered Pzq-treated metacercariae.The adult worms from Al-treated metacercariae were significantly bigger in size compared to the control group(P<0.05).Fecundity and body width were greater in adults from Mf-treated metacercariae compared to the control group(P<0.05).Conclusions:The larvicidal effects of these drugs were high efficacy to O.viverrini cercariae but lesser efficacy to metacercariae.It should be further studied with the eventual aim of developing a field-based control program.

Oral Amodiaquine, Artesunate and Artesunate Amodiaquine Combination Affects Open Field Behaviors and Spatial Memory in Healthy Swiss Mice

Effects of amodiaquine, artesunate and artesunate amodiaquine combination on open field novelty-induced behaviors and spatial memory in healthy mice were studied. Forty mice were used in the open field and fifty each in the radial arm maze and Y maze;mice were assigned into four or five groups of ten each, Group A served as control (distilled water), Groups B, C and D received artesunate (4 mg/kg), amodiaquine (10 mg/kg) and artesunate-amodiaquine combination (4 mg/kg and10 mg/kg) respectively, while Group E animals (for the cognition tests) were given scopolamine (2 mg/kg). Drugs and vehicle were administered orally for three days. Results were analysed by one way analysis of variance followed by a posthoc test. Results showed that artesunate and amodiaquine either in combination or administered singly caused a significant increase in open field novelty-induced horizontal locomotion and rearing. Grooming in the open field showed increments in the artesunate alone and artesunate amodiaquine groups while significant reductions in spatial memory were also seen in the cognition models used.

Effect of Artesunate vs Memantine in Aluminum Chloride Induced Model of Neurotoxicity in Rats

Alzheimer disease is one of the commonest neurological diseases which is characterized by amyloid plaques accumulation in multiple brain regions. This study investigated the potential neuroprotective effect of artesunate on aluminum induced neurotoxicity vs memantine in rats. 40 male albino Wistar rats were divided randomly into 4 groups as follow: Group 1 negative control, group 2 positive control group induced by ammonium chloride, group 3 rats treated by NH4Cl + artesunate solution, group 4 rats treated by NH4Cl + memantine S.C. spatial Memory and Learning were evaluated using Morris Water Maze (MWM) test. Malondialdehyde (MDA) and reduced glutathione (GSH) levels were measured in cerebral cortex tissue homogenate. Tumor necrosis factor-α (TNFα) and interleukin-1 beta (IL-1β) concentrations were measured in rat cerebral cortex tissue homogenate using rat enzyme linked immunosorbent assay (ELISA) kits. Real-time quantitative reverse transcription-polymerase chain reaction (Real-time qRT-PCR) for Caspase-3, Bcl-2 and iNOS gene expression was measured in rat cerebral cortex. Slices from cerebral cortex were studied by histopathological examination. Artesunate significantly decreased MDA level and inhibited iNOS, caspase and upregulated Bcl-2 gene expression in cerebral cortex. ART increased significantly antioxidant level GSH, and decreased significantly TNF-alpha and IL-B levels. It reduced significantly 1ry retention latency, 2ry retention latency and initial acquisition latency. It also improved brain histopathology and decreased amyloid plaque deposition. ART exerted neuroprotective effect through oxidative stress correction and enhancement of antiapoptotic markers in neuronal cells of the cerebral cortex.

Antimalarial drug artesunate affords protection against carrageenan induced acute inflammation in rat

Artesunate, an antimalarial drug, has been shown to inhibit the release of inflammatory mediators in various disease conditions. The present study was carried out to evaluate the anti-edematogenic effect of arte- sunate in the rat paw edema model. Inflammation was induced in the hind paw of rat by sub-plantar injection of 0.1 mL of 0.5% carrageenan and the paw volume was measured up to a fixed mark just before the injection and then after 3 h. The difference in two volumes gave a measure of edema formation. At 3h the level of TNF-α, PGE_(2) and myeloperoxidase were estimated in the inflamed paw tissue. Treatment of rats with single dose of artesunate at 50 and 150 mg/kg produced a dose-dependent inhibition in paw inflammation where a significant reduction in edema volume and mediator release was observed. Our study shows that by inhibiting the release of inflam- matory mediators artesunate affords protection against acute inflammation induced in the rat paw and suggests that it has a potential to be used in the treatment of inflammatory disease conditions.

Pharmacokinetics of Jiaotai pill self-microemulsion in insomnia rats

Objective:The pharmacokinetics and relative bioavailability of Jiaotai pill self-microemulsion were evaluated by investigating the blood concentration of Berberine,Coptisine,Palmatine and Jatrorrhizine in insomnia rats.Methods:Insomnia rat model was established by intraperitoneal injection of p-chlorophenylalanine(PCPA).The model rats were given Jiaotai pill self-microemulsion and Jiaotai pill suspension.The contents of Berberine,Coptisine,Palmatine and Jatrorrhizine in plasma at different times after administration were determined by UPLC-MS/MS,and calculate pharmacokinetic parameters.Results:Under the set chromatographic conditions,the linear relationship of the four components was good,and the precision,accuracy and stability meet the requirements of biological samples.After intragastric administration of Jiaotai pill self-microemulsion,The C_(max) of Berberine,Coptisine,Palmatine and Jatrorrhizine were(412.68±28.45),(68.65±3.92),(34.06±3.13),(40.60±1.22)ng/mL,and AUC_(0-∞)were(672.70±72.55),(146.04±25.01),(71.49±18.67),(72.25±9.54)ng·mL^(-1)·h^(-1),respectively.Compared with Jiaotai pill suspension,the Cmax,AUC_(0-t) and AUC_(0-∞)of the four components in insomnia rats were significantly increased(P<0.01).Conclusion:Jiaotai pill self-microemulsionl can promote the absorption of effective components in insomnia rats and improve its bioavailability.

Effects of Artesunate on Tracheal Smooth Muscle from the Guinea-pig

Artesunate is a derivative of qinghaosu, with a sesquiterpene structure. The specific action and the clinical uses of artesunate are on the preliminary stage. On the one hand, artesunate has specific action of both antiinflammation and antivirus, and also has protective effect on the pulmonary alveolar macrophages, which may be advantageous to the treatment of the airway non specific inflammation of asthma. On the other hand, qinghaosu has the activities to relax vascular smooth muscle and to cause hypotension. The expectorant action, the antitussive action and the antiasthmatic action of qinghaosu were reported. Artesunate may also have antiasthmatic activity, because the antimalarial potency of artesunate is stronger than that of qinghaosu, and Artesunate can block Ca 2+ influx by inhibiting calcium dependent chloride current. The main aims of this paper are to investigate the site, the mode, and the mechanism of artesunate action on isolated tracheal smooth muscle from the guinea pig. The isolated tracheal smooth muscle and isolated aortic strip circle were suspended in 10 ml Thornton and 5 ml Krebs solution gassed with 95% O 2+5% CO 2 at 37℃ respectively, and stretched with an initial tension of 1.5 g. After a stabilization period of over two hours, drug effects were plotted from cumulative doses. The tension changes were recorded by a forcedisplacement transducer connected to a two pen recorder (XWTD 264 made in Shanghai Dahua Apparatus Factory). Our experiments on isolated tracheal smooth muscle have demonstrated that artesunate is able to relax tracheal smooth muscle by its action on the tracheal smooth muscle cells. In concentrations ranging from 10 pmol·L 1 to 100 nmol·L 1 , artesunate can reduce the tone of the trachea in a concentration dependent manner. Its pD 2 is 8.76±0.74 (epithelium removed, n=6) and its potency is seventy four percent of isoprenaline′s. Artesunate is able to antagonize noncompetitively the effects of spasmogens like acetylcholine and histamine on trachea in a concentration dependent fashion, its pD′ 2 is 9.99±0.71 (n=8) and 11.69±0.53 (n=8), respectively. Timolol, a non selective blocker of the beta adrenergic receptors, does not inhibit the relaxant action of artesunate on trachea (n=6). Artesunate is also able to antagonize the constrictive effect on the trachea by KCl 100 mmol·L 1 in a concentration dependent manner, and its IC 50 is 0.81±0.67 nmol·L 1 (n=6). In the experiment of antagonizing the contractive action on trachea by acetylcholine 100 μmol·L 1 , artesunate was found to be more potent than nicardipine (n=7, P <0.01). Their inhibition rates (%) were 67.51±13.06% and 23.71±11.94% respectively. Nicardipine had no synergistic effect on the potency of artesunate (only increase the inhibition rate of artesunate from 65.71±11.06% to 73.94±11.78%, P>0.05, n=7). Artesunate had no effect on specific binding of 3H QNB on M 3 subtype of mAChR of salivary gland of rats (n=3). In the experiment of isolated aortic strip circle from the rats, artesunate did not block the intracellular Ca 2+ release, but partially inhibited Ca 2+ influx induced by phenylephrine 10 μmol·L 1 in a concentration dependent fashion. Its IC 50 was 1.64±0.38 mmol·L 1 (n=7). The intracellular Ca 2+ transient was determined using Fura 2 by fluorospectrophotometer (RF 5000 made in Japan). The excitation wavelengths were 340 and 380 nm and the emission wavelength was 510 nm. The intracellular Ca 2+ levels were calculated using the formula∶ [Ca 2+ ] i=Kd [(R t R min )/(R max R t)]×(Sf2/Sb2). In the cultured traheal smooth muscle cells, artesunate 100 μmol 1 had no effects on both intracellular Ca 2+ release and nonvoltagedepended Ca 2+ influx induced by cyclopiazonic acid (a Ca 2+ pump inhibitor of endoplasmic reticulum) 10 μmol·L 1 (n=3) (which was determined by using Fura 2). The cyclic AMP levels of the tracheal tissue were

Clinical Efficacy of Artemether-Lumefantrine Artesunate-Amodiaquine of Children from Three Chadian Provinces with Acute of Uncomplicated Falciparum malariae from 6 Months to 5 Years

Introduction: Artemisinin-based combination therapies are the first-line antimalarial drugs used to treat uncomplicated Plasmodium falciparum malaria in many endemic countries worldwide. In Chad, since the adoption of artemisinin-based combination therapy (ACTs) in its first-line treatment policy for uncomplicated malaria in 2005, Artemether-Lumefantrine and Artesunate-Amodiaquine have been used in many hospitals and health centers. The main objective of this study was to provide the baseline data of Artemether-Lumefantrine and Artesunate-Amodiaquine efficacy in three regions where many people suffered from malaria disease. Material and Methods: The baseline efficacy of two combination therapies was evaluated between January and April 2020 in Mosoro, Mondou and Dourbali Provinces in Chad. A two-arm single cohort study was conducted to assess the clinical efficacy of artesunate-amodiaquine and artemether-lumefantrine for the treatment of 1113 children aged from 6 to 59 months with uncomplicated Falciparum malariae diagnosed by thick blood smear examination, using the World Health Organization validated protocol. Results: On day 3, all patients in both groups had cleared parasitemia, after treatment, the patients presented a higher hemoglobin level in both groups artemether-lumefantrine (10.97 ± 1.39) and artesunate-amodiaquine (11.87 ± 1.81), respectively. On day 28, all patients had adequate clinical and parasitological responses with 99.82% of artesunate-amodiaquine and 99.10% of artemether-lumefantrine. Overall, both drugs were well tolerated at the clinical and biological level, no late parasitological failures have been recorded in artemether-lumefantrine and artesunate-amodiaquine groups, also both forms of Artemisinin-based combination therapy were still effective and safe in the treatment of uncomplicated P. falciparum malaria in Chad.