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Objective To establish an animal model by placing one end of PICC in the hepatic portal vein of a beagle dog and leaving the other end out of its body.Methods Six Beagle dogs were given respiration anesthesia through orotracheal intubation.An incision was made through the right rectus abdominalis to locate the superior mesenteric vein (SMA) and the main hepatic portal vein.The left branch of SMA was separated and cut to put PICC into the main hepatic portal vein before being ligated and fixed.The other end of PICC was elicited through the right abdominal wall and passed beneath the skin to the back neck and fastened in case of movement.Results The anesthetic effect was good and all the operations were successful.The mean operation time was about an hour and the mean blood loss was about 15 ml.The incision healed 5-7 d after operation.Conclusion The establishment of the model can improve the effects of liver-targeting drugs,which can cut down the dosage,lower the cost of treatment and experiment and reduce the adverse effect of medicines.Through PICC,we can directly draw blood from the hepatic portal vein to measure the blood concentration before the first pass elimination.Then according to the concentration,we can calculate the absorption rate in the gastrointestinal tract,which can facilitate related experimental studies.
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Objective@#To determine the scavenging effect and the change of metabolism of paraquat (PQ) using hemoperfusion (HP) once and twice within 12 hours after intoxication and explore the better scheme of HP.@*Methods@#18 beagles were randomly divided into 3 groups. Single HP group, Double HP group and Control group. Peripheral veins blood was collected at different times within 48 hours after exposure in each group. Toxin concentration was measured, analyzed and compared among 3 groups.@*Results@#6 hours after exposure, Single HP group and Double HP group has finished the first HP treatment, and the concentration of PQ was lower than that of the control group, the difference was statistically significant (P<0.05) . 10 hours after exposure, there was no statistical difference of toxin concentration among 3 groups (P>0.05) . 12 hours after exposure, Double HP group has finished the second HP treatment, the concentration of PQ was significantly lower than that of Single HP group and Control group (P<0.05) . 24 hours and 48 hours after exposure, there was no statistical difference of toxin concentration among 3 groups (P>0.05) . Statistical difference were not observed in toxicokinetical parameters among 3 groups (P>0.05) .@*Conclusion@#HP treatment once and twice within 12 hours after intoxication could effectively reduce the toxin concentration in the peripheral veins blood after HP for about 4 hours, then the toxin concentration would return to the same level as Control group quickly. It was suggested that at the beginning of poisoning, HP treatment once or twice could not significantly change the metabolism of paraquat.
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Objective To study the adsorption effect of activated charcoal suspension on paraquat (PQ) in gastrointestinal tract of beagles exposed to PQ.Methods Twenty healthy male beagles were randomly divided into experimental group and control group,with 6 beagles in each group.20％ PQ solution (a dose of 30 mg/kg) was prescribed through stomach for beagles in both groups.After exposure to PQ for 30 minutes,the beagles in experimental group were given activated charcoal suspension (1.0 g/kg of type Ⅰ activated charcoal powder mixed with 100 mL of normal saline) by gavage,while the control group was only given equal volume of normal saline.After exposure to PQ for 10 minutes,30 minutes,and 1,2,4,8,12,24,and 48 hours,blood was collected from hepatic portal veins and peripheral veins to detect the PQ concentration change in the plasma.The toxicokinetics software DAS 2.1.1 was applied to analyze PQ concentration and compare the change in toxicokinetics parameters between the both groups.The change in vital signs including heart rate (HR),respiratory rate (RR) and pulse oxygen saturation (SpO2) was dynamically monitored 10 minutes before exposure,4 hours and each day from the 1st to the 7th day after exposure.Results After exposure to PQ,the poison concentration in the plasma of hepatic portal veins and peripheral veins in the control group rose quickly and reached peak 4 hours later.It fell quickly at first,and fell slowly 8 hours later.But in the experimental group,the increase rate to the peak was significantly slow.Besides,PQ peak fell more obviously than that in the control group and it was about 50％ of the control group (μg/L:123.50 ± 11.67 vs.255.18 ± 12.29 in blood from hepatic portal veins,122.35± 11.72 vs.250.86± 11.15 in blood from peripheral veins).After 8 hours it fell much more quickly than that of the control group.After exposure to PQ for 48 hours,PQ concentration in the plasma was still lower than that of the control group (μg/L:0.53 ± 0.18 vs.15.98 ± 5.58 in blood from hepatic portal veins,0.31 ± 0.01 vs.15.03 ± 4.82 in blood from peripheral veins,both P ＜ 0.01).With the toxicokinetics analysis,compared with the control group,the maximum concentration (Cmax) and area under the curve (AUC) of PQ in the plasma of hepatic portal veins and peripheral veins in the experimental group were significantly decreased [Cmax (μg/L):125.07 ± 9.49 vs.255.18 ± 12.29 in blood from hepatic portal veins,123.38 ± 9.52 vs.250.86 ± 11.15 in blood from peripheral veins;AUC (mg· L-1· h-1):1.6±0.2vs.3.3 ± 0.4 in blood from hepatic portal veins,1.5 ± 0.2 vs.3.2 ± 0.3 in blood from peripheral veins],time to the peak (Tmax) of PQ was slowed (hours:5.3 ± 1.9 vs.4.0 ± 0.0 in blood from hepatic portal veins,4.7 ± 1.5 vs.4.0 ± 0.0 in blood from peripheral veins),and PQ plasma half-life (t1/2) and mean retention time (MRT) were significantly shortened [t1/2 (hours):3.8 ± 1.2 vs.15.4± 3.7 in blood from hepatic portal veins,3.5 ± 1.0 vs.15.5 ± 2.7 in blood from peripheral veins;MRT (hours):8.0± 1.5 vs.13.4± 1.2 in blood from hepatic portal veins,7.6± 1.3 vs.13.3± 1.2 in blood from peripheral veins;all P ＜ 0.01].After exposure to PQ,HR and RR in both the experimental group and the control group increased and reached to the peak about the 4th day and then the increase rate began to slow down gradually;SpO2slowed down gradually and reached to the valley about the 4th day and then it began to recover,but the change range of vital signs in the experimental group was smaller than that of the control group,and the parameters were significantly better than those of control group [4-day HR (bpm):134.50±3.00 vs.142.00±6.43,4-day RR (times/min):31.00±0.58 vs.34.33±0.94,4-day SpO2:0.900±0.006 vs.0.873±0.005,all P ＜ 0.05].Conclusion Activated charcoal administrated at 30 minutes after PQ poisoning can slow down the increase rate of PQ concentration in the plasma,decrease the peak concentration and has less influence on vital signs in beagles.
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Objective To establish acute hepatotoxic model induced by Amanita exitialis and to study the characteristics of acute toxic liver failure induced by mushrooms containing peptide toxins,in hope for providing some help to experimental research on poisoning induced by mushrooms containing peptide toxins.Methods UPLC-MS/MS (Ultra performance liquid chromatography-tandem mass spectrometry) method was used to detect peptide toxins in Amanita exitialis.To establish acute toxic liver hepatic failure model,the beagles were fed with 60 mg/kg of lyophilized powder of Amanita exitialis fungus which encapsulated in starch capsules.Toxic sighs were observed,coagulation function,hepatic and renal function,liver histopathological morphology,peptide toxin concentration in plasma and urine were detected during the experiment.Results Total peptide toxins in Amanita exitialis was (3 482.6 ± 124.94) mg/ kg.All the beagles had toxic signs including vomiting and diarrhea in 12-48 h after ingestion.On 24 h after ingestion,the beagles' ALT,AST,TBIL,ALP,PT and APTT levels increased obviously.On 36 h after ingestion,the beagles' ALT,AST,PT and APTT values reached their peaks (ALT:283.2 ± 112.9 Kallmann unit;AST:223.9 ±93.8 Kallmann units;PT:132.9 ± 152.6 s;APTT:131.4 ± 153.9 s).On 48 h after ingestion,the beagles' TBIL and ALP levels reached their peaks (TBIL:23.3 ± 14.6 mol/L;ALP:274.5 ± 115.5 U/L).The beagles' TBIL,TP and APTT returned to normal 1 week after ingestion,their ALT,AST and ALP levels returned to normal 3 weeks after ingestion.Three dogs died during 24-72 h after ingestion.Liver histopathological morphology study showed hemorrhagic necrosis of hepatocytes.Peptide toxins can be detected in plasma within 24 h after ingestion.Peptide toxins can be detected in urine within 96 h after ingestion.Conclusion Amanita peptide toxins can cause hemorrhagic necrosis of liver cells and lead to acute liver failure.This model is consistent with clinical pathophysiological process of acute toxic liver failure induced by mushrooms containing peptide toxins,and it can be applied to the study of diagnosis and treatment of poisoning induced by mushrooms containing peptide toxins.
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Tongmai granule (TM) is composed of Puerariae Lobatae Radix (Gegen), Salviae Miltiorrhizae Radix et Rhizoma(Danshen) and Chuanxiong Rhizoma(Chuanxiong). It has been used to treat ischemic cardio-cerebrovascular diseases for decades. For the purpose of elucidating its pharmacodynamic material foundation, the absorption and pharmacokinetic property of TM were investigated in acute myocardial ischemic model beagles. All serum samples were extracted before analysis with ethyl acetate after being acidified by hydrochloric acid. Under negative ESI detection mode, the chromatographic separation was carried out with monolithic C₁₈ column for gradient elution. A simultaneous quantitative analysis was made on 15 polyphenols, including 8 from Gegen, 5 from Danshen and 2 from Chuanxiong, in 8.5 min. The validation result demonstrated the specificity, accuracy and precision of the method in line with the bioanalysis requirements. After TM solution was administrated to acute myocardial ischemic model beagles through duodenum injection, serum samples were collected after 6 h. The quantitative detection proved the prompt absorption of TM, all of the components were detectable in the blood samples 5 min later, and reached peak respectively at 0.18-3.83 h after administration. The components presented large variabilities. The most components were exposed in serum with puerarin and salvianic acid A, followed by 3'-methoxypuerarin, mirificin, and 3'-hydroxypuerarin. The study proves puerarin and salvianic acid A are dominating active components of TM in acute myocardial ischemic model beagles.
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Objective To observe the effect of 7 kinds of Chinese herbs(Pericarpium Arecae, Folium Sennae, Fructus Tsaoko, Fructus Amomi, Rhizoma Atractylodis Macrocephalae, Fructus Aurantii, Fructus Aurantii I mmaturus), and 2 kinds of active compounds (bornyl acetate, synephrine) of Chinese herbals on gastrointestinal motility of chronic experimental beagle model. Methods Six beagles were used for inducing chronic experimental model. The beagles’antrum, duodenum, jejunum, ileum and colon were implanted with strain gauges to record canine gastrointestinal motility. Gastric fistula was set up for the intake of Chinese medicine decoction, and the external jugular vein catheter was made for intravenous administration. After modeling, the fasting gastrointestinal motility features of the beagles were observed for 120 min every day, and for 7 continuous days. From the 7th day after modeling, fasting gastrointestinal motility before medication was recorded as fundamental control, and when the interdigestive migrating motor complex (MMC) I phase arrived, the gastrointestinal motilities were sequently recorded after treatment with the 7 kinds of herbs(gavage of 200 mL of the decoction of each herb through gastric fistula), normal saline(200 mL), bornyl acetate(active ingredient of Fructus Aurantii and Fructus Aurantii Immaturus, intravenous injection), and synephrine (active ingredient of Fructus Amomi, intravenous injection). All of the animals were treated with only one kind of Chinese herb one day , and the observation of each herb lasted for 2 continuous days. MMC cycle, frequency of contraction, sum of contraction, amplitude of contraction, average of amplitude, and motor index (MI) were observed by strain gauges. Results On postoperative day 1-6, the animals had gastrointestinal hypomotility and no MMC cycle was recorded. On postoperative day 7, the canine antrum, duodenum, jejunum and ileum showed typical MMC cycle while the colon had irregular MMC cycle at fasting interdigestive period. Compared with the gastrointestinal motilities before treatment and those of the normal saline group, the number of contraction waves and MI were significantly increased in the antrum, duodenum, jejunum, ileum and colon after intervention with Pericarpium Arecae and bornyl acetate, were increased in the duodenum, jejunum, ileum and colon after treatment with Folium Sennae and Fructus Tsaoko (P<0.05), and were also enhanced in the antrum and duodenum after administration of Fructus Amomi(P < 0.05). Treatment with Fructus Aurantii induced the decrease of MI in the jejunum, Fructus Aurantii Immaturus decreased MI of the jejunum and colon, and synephrine decreased the number of antrum contraction waves and MI of the antrum and jejunum(P < 0.05). Conclusion The chronic experimental model is effective for the screening of Chinese herbs for improving gastrointestinal motility. Pericarpium Arecae, Folium Sennae, Fructus Tsaoko, Fructus Amomi, and bornyl acetate can increase gastrointestinal motility, while Fructus Aurantii, Fructus Aurantii Immaturus and synephrine can inhibit the gastrointestinal movement, and Rhizoma Atractylodis Macrocephalae has no effect on gastrointestinal motility.
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OBJECTIVE: To study the effect of isosorbide-5-mononitrate (5-ISMN) on ivabradine hydrochloride pharmacokinetics in Beagles.
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@#ObjectiveToexplorethemethodtoestablishanimalmodelofearlyosteonecrosisofthefemoralhead(ONFH)inducedby steroid.Methods20healthymaleBeagleswererandomlydividedintocontrolgroupandexperimentalgroupwith10dogsineachgroup. Theexperimentalgroupwasinjectedlipopolysaccharide10μg/kgandmethylprednisolone20mg/kgfor3daysconsecutively.Thecontrol groupwasinjectednormalsaline.2monthsand4monthsafteradministration,bothgroupswereperformedmagneticresonanceimaging (MRI).5animalsweresacrificedrespectivelyat2monthsand4monthsafteradministrationineachgroup,andbilateralfemoralheadspeci-menswereobtainedtoperformhistologicalexamination.Plasmaprothrombintime(PT),activatedpartialthromboplastintime(APTT),anti-thrombinIII(AT-III)weretestedbeforeand24hafteradministration.ResultsIntheexperimentalgroup,thepathologicalresultsshowed thattherewere4ONFH2monthsand6ONFH4monthsafteradministrationandMRIdidnotshowanyabnormality.Comparedwiththe controlgroup,thePT,APTT,AT-IIIintheexperimentgroupshortenedsignificantlyafteradministration(P<0.001).ConclusionModified steroid-inducedmethodcanestablishtheanimalmodelofearlyONFH.Hypercoagulationandlowfibrinolysismaybethereasonofste-roid-inducedosteonecrosis.
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Objective To construct and identify retroviral-mediated short hairpin RNA ( shRNA ) expression vectors of ERβ419, and explore ERβ419 unknown biological function in beagles in future.Methods To screen out the most effective gene silencing sequence of beagle ERβ419 mRNA using qRT-PCR and Western Blot assays, imitate beagle estrogen target cells.Results qRT-PCR results showed, ERβ419-shRNA1 ( P <0.01 ) and ERβ419-shRNA3 ( P <0.01)differed significantly, Western Blot result as same as qRT-PCR,ERβ419-shRNA3 is the best choice.Conclusion Beagles ERβ419-shRNA3 retrain most effectively target gene repression. It is applied to explore ERβ419 unknown biological function in beagles reproductive system, and to prevent and treat beagles reproductive function diseases.
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Objective To quantitative the changing information of estrogen receptor βgene which was in tissue and organ of sex gland during oestrus and dioestrus of Beagles, and to show the different expression situation of hypothalamus-pituitary-gonad axis during oestrus and dioestrus, and providing the basic of theory to research deeply the mechanism of heat of Beagles. Methods As the key gene in regulation reproduction, ERβgene is located in hypothalamus-pituitary-gonad axis, so using Beagles which was in oestrus and dioestrus, and extract the RNA from hypothalamus、pituitary、ovary and uterus respectively,after reverse transcription we detected the expression of ERβgene by real-time quantitative PCR.Results The expression of ERβgene mRNA from ovary、uterus、pituitary、hypothalamus of Beagles which was in dioestrus was 0.35 times, 0.17 times, 0.44 times and 0.43 times than the expression of ERβgene mRNA from ovary, uterus, pituitary, hypothalamus of Beagles which was in oestrus.Conclusion The expression of ERβgene was up-regulation in hypothalamus-pituitary-ovary axis of Beagles which was in oestrus.
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Objective To screen the splicing isoforms of estrogen receptor βin the Beagle hypothalamic -pituitary -gonadal axis.Methods For ERβmRNA CDS sequence of eight exons, primers were designed confined to the CDS sequences of two sequential exons.Beagle hypothalamus, pituitary, ovary and uterus tissue cDNA were used as template, and corresponding sequences were amplified by PCR.PCR products were sequenced and aligned in the NCBI web site.The correct gene was then analyzed with DNAMAN comparative analysis software and handwork checking up, thus got the ERβsplicing isoforms of Beagle. Results Four beagle ER beta splicing isomers were obtained:exon 4 complete skipping ER βisomer (300 bp missing), two kind of Beagle ERβisoforms with partial exon 4 and partial exon 5 complicated missing (isoformⅠ334 bp missing and isoformⅡ265 bp missing), and exon 7 complete missing ERβsplicing isoforms (181 bp missing).Exon 4 complete skipping and exon 7 complete missing isomers had been obtained full length coding sequence, and the other two splicing isomers were partial coding sequence.Conclusion This project gained four ERβsplicing isomers of Beagle, and that will lay an important foundation for further study of their roles in the Beagle reproductive regulation mechanism.
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@#ObjectiveTo assess the safety of intramyocardium injection of sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) to rescue chronic heart failure beagles. MethodsThe heart failure beagles were assessed with myocardium enzymology, cAMP content of myocardium, myocardial oxygen consumption, arrhythmia, systemic inflammation factors, and the function of liver and kidney after SERCA2a gene transferred. ResultsThere were no increase of cAMP content, no more arrhythmia, myocardial oxygen consumption, no apparente systemic inflammation, and no injures to hepatic and renal function. ConclusionOverexpression of SERCA2a by intramyocardium injection is relatively safe.
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Objective To evaluate the effects of combined administration of recombinant human interleukin-11(rhIL-11),recombinant human G-CSF(rhG-CSF)and recombinant human interleukin-2 (rhIL-2)on acute radiation sickness(ARS)beagles.Methods Sixteen beagle were irradiated with 4.5 Gy60 Co γ-rays to establish ARS models,and were divided into irradiation control group,supportive care group and combined cytokines treatment group.After irradiation irradiation control group was given no treatment,the dogs in supportive care group received purely symptomatic treatment,while combined cytokines treatment group received rhIL-11 50μg/(kg·d)and rbG-CSF 10μg/(kg·d)subcutaneously(0-14 d)and rhIL-2 1×1 06 U/d(29-43 d)besides symptomatic treatment.Manifestation and characteristics of ARS beagles were observed,and the survival time were recorded.At last,post-mortem examination and histological examination were performed.Results All animals underwent nausea,diarrhea and fever.After irradiation,all animals in irradiation control group died in two weeks,and the mean survival time was 12.7 d,while only one died at 33 d in supportive care group.All dogs in combined cytokine group survived at 45 day after exposure,and their haematopoiesis and gastrointestinal tract were recovered.Conclusions Combination of rhIL-11 + rhG-CSF + rhIL-2 treatment could be significantly effective on ARS beagles irradiated by 4.5 Gy60 Co γ-rays,which could accelerate injured haemotopoiesis and intestinal tract recovery,increase the survival rate and improve the life quality of animals.
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Objectivc To observe the therapeutic effects of combined cytokines on hematopoietic injuries induced by 4.5 Gy60 Co γ-rays irradiation in beagles,and to provide experimental evidences for the clinical treatment of extremely severe myeloid acute radiation sickness(ARS).Methods 16 beagles were given 4.5 Gy60 Co γ-rays total body irradiation,and then randomly assigned into irradiation control group,supportive care group and cytokines group.In addition to supportive care,recombinant human granulocyte colony-stimulating factor (rhG-CSF),recombinant human interleukin-11(rhIL-11)and recombinant human interleukin-2(rhIL-2)were administered subcutaneouly to dogs in cytokines group.Peripheral blood hemogram was examined once every two days.Bone marrow and peripheral blood were collected to proceed colony cultivation 4 d pre-irradiation and 1 and 45 d post-irradiation.Conventional histopathological sections of sternum were prepared to observe the histomorphology changes. Results After irradiation,the population of all kinds of cells in peripheral blood declined sharply.WBC nadir Was elevated(1.04×109/L,but 0.28×109/L and 0.68×109/L for the irradiation control group and the supportive care group separately),the duration of thrombocytopenia was shortened (24 days,but 33 days for the supportive care groug) and red blood cell counts were maintained in the range of normal values after cytokincs treatment in combination.The colony forming efficiency of haemopoietic stem cells(HSCs)in bone marrow and peripheral blood decreased obviously 1 d post irradiation,but recovered to the level of that before irradiation 45 d post irradiation after supportive care and cytokines treatment.Hematopoietic cells disappeared in bone marrow of animals in irradiation control group,but hematopoietic functions were recovered after cytokines were administrated.Conclusions RhG-CSF.rhIL-11 and rhIL-2 used in combination could elevate WBC nadir,accelerate the recovery of leukocytes,platelets and red blood cells and promote the proliferation,differentiation and maturity of HSPCs left in the body after 4.5 Gy γ-rays total body irradiation,eventually restore the hematopoietic function.Hence,combination of rhG-CSF,rhIL-11 and rhIL-2 could serve as better therapeutic strategy to treat extremely severe myeloid ARS.
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Objective To investigate the mechanism of treatment of granulocyte colony-stimulating factor(rhG-CSF),recombinant human interleukin-11(rhIL-11)and recombinant human interleukin-2 (rhIL-2)on hematopoietic injuries induced by 4.5 Gy60 Coγ-ray irradiation in beagles,and to provide experimental evidence for the clinical treatment of extremely severe myeloid acute radiation sickness (ARS).Methods Sixteen beagle dogs were given 4.5 Gy60 Co γ-ray total body irradiation(TBI),then randomly assigned into irradiation control group,supportive care group or cytokines+supportive care (abbreviated as cytokines)group.In addition to supportive care,rhG-CSF,rhlL-11 and rhIL-2 were administered subcutaneously to treat dogs in cytokines group.The percentage of CD34+cells,cell cycle and apoptosis of nucleated cells in peripheral blood were examined by Flow cytometry.Results After 4.5 Gy 60 Co γ-ray irradiation,the CD34+cells in peripheral blood declined obviously(61.3%and 52.1% of baseline for irradiation control and supportive care group separately).The cell proportion of nucleated cells in Go/G1 phase was increased notably(99.27% and 99.49% respectively).The rate of apoptosis(26.93% and 21.29% separately)and necrosis(3.27% and 4.14%,respectively)of nucleated cells were elevated significantly when compared with values before irradiation(P<0.05) 1 d post irradiation.When beagles were treated with cytokines and supportive care,the CD34+cells in peripheral blood were markedly increased(135.6% of baseline).The effect of G0/G1 phase blockage of nucleated cells became more serious(99.71%).The rate of apoptosis(5.66%)and necrosis(1.60%)of nucleated cells were significantly lower than that of irradiation control and supportive care groups 1 d after exposure.Conclusions Cytokines maybe mobilize CD34+cells in bone marrow to peripheral blood,indce cell cycle block at G0/G1 phase and reduce apoptosis,and eventually cure hematopoieticinjuries induced by irradiation.
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[Objective] To investigate the renal histological parameters in SD rats and beagles and to supply evidence for renal toxicological and pathological diagnosis. [Methods] Forty SD adult rats and 20 adult beagles (half males and half females) , were respectively executed at 12 weeks and 12 months old. Then, kidney tissue was fixed in 10% neutral formalin, embedded in paraffin, made into slices at 5?m and stained by HE. The larger diameter and shorter diameter of glomerulus in the renal cortex and outer stripe of rats and beagles were determined by Spot Advanced image analysis software. The differences of the above parameters appearing in different species and genders were compared. [Results] The larger diameter was about 96 ?m, and the shorter diameter 76 ?m in the renal cortex of the beagles, and the larger was about 120?m and the shorter 94 ?m in the outer stripe of the beagles. The differences of the larger and shorter diameters were insignificant between the male and female beagles, but the larger and shorter diameters in the outer stripe were longer than those in the cortex. The larger diameter and shorter diameter were about 87 ?m and 64 ?m respectively in the renal cortex of SD rats, and the larger was about 88 ?m and the shorter 64 ?m in the outer stripe of SD rats. The differences of the larger and shorter diameters were insignificant between the male and female beagles, as well as in the outer stripe and the cortex. [Conclusion] The nephron in the outer stripe is larger than that in the cortex of beagles but no difference of the nephron exists in SD rats. The nephron in beagles is larger than that in SD rats, and there is no difference in the nephron between male and female beagles and SD rats.