Experimental nerve transfer model in the neonatal rat

Clinically,peripheral nerve reconstructions in neonates are most frequently applied in brachial plexus birth injuries.Most surgical concepts,however,have investigated nerve reconstructions in adult animal models.The immature neuromuscular system reacts differently to the effects of nerve lesion and surgery and is poorly investigated due to the lack of reliable experimental models.Here,we describe an experimental forelimb model in the neonatal rat,to study these effects on both the peripheral and central nervous systems.Within 24 hours after birth,three groups were prepared:In the nerve transfer group,a lesion of the musculocutaneous nerve was reconstructed by selectively transferring the ulnar nerve.In the negative control group,the musculocutaneous nerve was divided and not reconstructed and in the positive control group,a sham surgery was performed.The animal's ability to adapt to nerve lesions and progressive improvement over time were depict by the Bertelli test,which observes the development of grooming.Twelve weeks postoperatively,animals were fully matured and the nerve transfer successfully reinnervated their target muscles,which was indicated by muscle force,muscle weight,and cross sectional area evaluation.On the contrary,no spontaneous regeneration was found in the negative control group.In the positive control group,reference values were established.Retrograde labeling indicated that the motoneuron pool of the ulnar nerve was reduced following nerve transfer.Due to this post-axotomy motoneuron death,a diminished amount of motoneurons reinnervated the biceps muscle in the nerve transfer group,when compared to the native motoneuron pool of the musculocutaneous nerve.These findings indicate that the immature neuromuscular system behaves profoundly different than similar lesions in adult rats and explains reduced muscle force.Ultimately,pathophysiologic adaptations are inevitable.The maturing neuromuscular system,however,utilizes neonatal capacity of regeneration and seizes a variety of compensation mechanism to restore a functional extremity.The above described neonatal rat model demonstrates a constant anatomy,suitable for nerve transfers and allows all standard neuromuscular analyses.Hence,detailed investigations on the pathophysiological changes and subsequent effects of trauma on the various levels within the neuromuscular system as well as neural reorganization of the neonatal rat may be elucidated.This study was approved by the Ethics Committee of the Medical University of Vienna and the Austrian Ministry for Research and Science(BMWF-66.009/0187-WF/V/3 b/2015)on March 20,2015.

A Biochemical Study on Combined Treatment of Experimental Silicosis with Tetradrine-PVNO and Tetradrine-QOHP in Rats

A better understanding is needed to explain the mechanism of therapeutic effect of combined use of tetradrine-PVNO and tetradrine-QOHP which play very important roles in treatment of silicosis. Blood prolidase (PLD), monoamine oxidase (MAO) and plasminogen (PLG) in silicotic rats after treatment with tetradrine-PVNO or tetradrine-QOHP were measured. The values obtained were compared with the untreated silicotic rats. It was found that the silicotic rats that received tetradrine-PVNO showed significant increase in PLD and decrease in PLG, but no significant change in MAO. The PLD in plasma of silicotic rats that received tetradrine-QOHP were elevated significantly, but PLG and MAO did not change appreciably. These findings suggest that the combined use of tetradrine-PVNO and tetradrine-QOHP can accelerate the degradation of collagen in silicotic rats

Experimental observation of rat's early-stage fracture healing with different kinds of nerve injuries

To investigate the impact of different kinds of nerve injuries of early-stage fracture healing.Methods Three groups of rats were included in the experiment among which group 1 was inflicted with femoral fracture and T10 spinal cord transsection (SCI),group 2 was inflicted with femoral and peripheral nerve resection (PNR),and group 3 with simple femoral fracture as control group.Two weeks after operation the femoral bones were collected for X-ray checking and 2 more weeks later X-ray checking was performed again followed by pathomorphologic exams.Results X-ray result showed no massive calluses in the bones in the 2nd week postoperatively,while in the 4th week,callus appeared with larger size in group 3 than that of group 1 and with smaller size than that of group 2.It was the same with the result of pathomorphologic examining.Cortical bone bridges between fracture point and osteiod were also found in group 2 and there were less normal blood vessels and worse bone remodeling than that of group 3.There were relatively immature calluses with more fibroblast-like cells and disordered bone structure in group 2.Group 3 showed normal healing process and callus structure.Conclusion Early-stage bone fracture healing can be influenced significantly by different kinds of nerve injuries.6 refs,6 figs.

Alleviated mucosal and neuronal damage in a rat model of Crohn's disease

AIM:To establish a rat model suitable to investigate the repetitive relapsing inflammations(RRI)characteristic to Crohn’s disease.METHODS:Colitis was induced by 2,4,6-trinitrobenzenesulfonic acid(TNBS).RRI were mimicked by repeating administrations of TNBS.Tissue samples were taken from control,once,twice and three times treated rats from the inflamed and adjacent non-inflamed colonic segments at different timepoints during the acute intestinal inflammation.The means of the ulcerated area were measured to evaluate the macroscopic mu-cosal damage.The density of myenteric neurons was determined on whole mounts by Hu C/Hu D immunohistochemistry.Heme oxygenase-1(HO-1)expression was evaluated by molecular biological techniques.RESULTS:TNBS-treated rats displayed severe colitis,but the mortality was negligible,and an increase of body weight was characteristic throughout the experimental period.The widespread loss of myenteric neurons,and marked but transient HO-1 up-regulation were demonstrated after the first TNBS administration.After repeated doses the length of the recovery time and extent of the ulcerous colonic segments were markedly decreased,and the neuronal loss was on a smaller scale and was limited to the inflamed area.HO-1 m RNA level was notably greater than after a single dose and overexpression was sustained throughout the timepoints examined.Nevertheless,the HO-1protein up-regulation after the second TNBS treatment proved to be transient.Following the third treatment HO-1 protein expression could not be detected.CONCLUSION:Experimentally provoked RRI may exert a protective preconditioning effect against the mucosal and neuronal damage.The persistent up-regulation of HO-1 m RNA expression may correlate with this.

High yield reproducible rat model recapitulating human Barrett's carcinogenesis

AIM To efficiently replicate the biology and pathogenesis of human esophageal adenocarcinoma(EAC) using the modified Levrat model of end-to-side esophagojejunostomy. METHODS End-to-side esophagojejunostomy was performed on rats to induce gastroduodenoesophageal reflux to develop EAC. Animals were randomly selected and serially euthanized at 10(n = 6),17(n = 8),24(n = 9),31(n = 6),38(n = 6),and 40(n = 6) wk postoperatively. The esophagi were harvested for downstream histopathology and gene expression. Histological evaluation wascompleted to determine respective rates of carcinogenic development. Quantitative reverse transcriptionpolymerase chain reaction was performed to determine gene expression levels of MUC2,CK19,and CK20,and results were compared to determine significant differences throughout disease progression stages.RESULTS The overall study mortality was 15%. Causes of mortality included anastomotic leak,gastrointestinal hemorrhage,stomach ulcer perforation,respiratory infection secondary to aspiration,and obstruction due to tumor or late anastomotic stricture. 10 wk following surgery,100% of animals presented with esophagitis. Barrett's esophagus(BE) was first observed at 10 wk,and was present in 100% of animals by 17 wk. Dysplasia was confirmed in 87.5% of animals at 17 wk,and increased to 100% by 31 wk. EAC was first observed in 44.4% of animals at 24 wk and increased to 100% by 40 wk. In addition,two animals at 38-40 wk post-surgery had confirmed macro-metastases in the lung/liver and small intestine,respectively. MUC2 gene expression was progressively down-regulated from BE to dysplasia to EAC. Both CK19 and CK20 gene expression significantly increased in a stepwise manner from esophagitis to EAC. CONCLUSION Esophagojejunostomy was successfully replicated in rats with low mortality and a high tumor burden,which may facilitate broader adoption to study EAC development,progression,and therapeutics.

Establishment of rat experiment model of chronic allograft nephropathy

Objective To summarize experience of establishing stable rat model ot chronic allograft nephropathy. Methods We used Fisher rats as donors and Lewis rats as recipients. After left kidney of donor perfused in situ under hypothermic condition,left renal vein,abdominal aorta and bladder flap of donor was anastomosed with

Juvenile ferric iron prevents microbiota dysbiosis and colitis in adult rodents

AIM: To assess whether juvenile chronic ferric iron ingestion limit colitis and dysbiosis at adulthood in rats and mice. METHODS: Two sets of experiments were designed. In the first set, recently weaned mice were either orally administered ferrous (Fe2+) iron salt or ferric (Fe3+) microencapsulated iron for 6 wk. The last week of experiments trinitrobenzene sulfonic acid (TNBS) colitis was induced. In the second set, juvenile rats received the microencapsulated ferric iron for 6 wk and were also submitted to TNBS colitis during the last week of experiments. In both sets of experiments, animals were sacrificed 7 d after TNBS instillation. Severity of the inflammation was assessed by scoring macroscopic lesions and quantifying colonic myeloperoxidase (MPO) activity. Alteration of the microflora profile was estimated usingquantitative polymerase chain reaction (qPCR) by measuring the evolution of total caecal microflora, Bacteroidetes, Firmicutes and enterobacteria. RESULTS: Neither ferrous nor ferric iron daily exposures at the juvenile period result in any effect in control animals at adulthood although ferrous iron repeated administration in infancy limited weight gain. Ferrous iron was unable to limit the experimental colitis (1.71 ± 0.27 MPO U/mg proteinvs 2.47 ± 0.22 MPO U/mg protein in colitic mice). In contrast, ferric iron significantly prevented the increase of MPO activity (1.64 ± 0.14 MPO U/mg protein) in TNBS-induced colitis. Moreover, this positive effect was observed at both the doses of ferric iron used (75 and 150 mg/kg per day po - 6 wk). In the study we also compared, in both rats and mice, the consequences of chronic repeated low level exposure to ferric iron (75 mg/kg per day po - 6 wk) on TNBS-induced colitis and its related dysbiosis. We confirmed that ferric iron limited the TNBS-induced increase of MPO activity in both the rodent species. Furthermore, we assessed the ferric iron incidence on TNBS-induced intestinal microbiota dysbiosis. At first, we needed to optimize the isolation and quantify DNA copy numbers using standard curves to perform by qPCR this interspecies comparison. Using this approach, we determined that total microflora was similar in control rats and mice and was mainly composed of Firmicutes and Bacteroidetes at a ratio of 10/1. Ferric juvenile administration did not modify the microflora profile in control animals. Total microflora numbers remained unchanged whichever experimental conditions studied. Following TNBS-induced colitis, the Firmicutes/Bacteroidetes ratio was altered resulting in a decrease of the Firmicutes numbers and an increase of the Bacteroidetes numbers typical of a gut inflammatory reaction. In parallel, the subdominant population, the enterobacteria was also increased. However, ferric iron supplementation for the juvenile period prevented the increase of Bacteroidetes and of enterobacteria numbers consecutive to the colitis in both the studied species at adulthood.CONCLUSION: Rats and mice juvenile chronic ferric iron ingestion prevents colitis and dysbiosis at adulthood as assessed by the first interspecies comparison.

Evaluation of the biological toxicity of fluorine in Antarctic krill

Antarctic krill is a potentially nutritious food source for humans, but lfuorine （F） toxicity is a matter of concern. To evaluate the toxicity of F in Antarctic krill, 30 Wistar rats were divided into three groups with different dietary regimens：a control group, a krill treatment group （150 mg＆#183;kg-1 F）, and a sodium lfuoride （NaF） treatment group （150 mg＆#183;kg-1 F）. After three months, F concentrations in feces, plasma, and bone were determined, and the degree of dental and skeletal lfuorosis was assessed. The F concentrations in plasma and bone from the krill treatment group were 0.167 0＆#177;0.020 4 mg.L-1 and 2 709.8＆#177;301.9 mg＆#183;kg-1, respectively, compared with 0.043 8＆#177;0.005 5 mg＆#183;L-1 and 442.4＆#177;60.7 mg＆#183;kg-1, respectively, in samples from the control group. Concentrations of F in plasma and bone in the krill treatment group were higher than in the control group, but lower than in the NaF treatment group. The degree of dental lfuorosis in the krill treatment group was moderate, compared with severe in the NaF treatment group and normal in the control group. The degree of skeletal lfuorosis did not change signiifcantly in any group. These results showed that the toxicity of F in Antarctic krill was lower than for an equivalent concentration of F in NaF, but it was toxic for rats consuming krill in large quantities. To conclude, we discuss possible reasons for the reduced toxicity of F in Antarctic krill. The present study provides a direct toxicological reference for the consideration of Antarctic krill for human consumption.

Biomechanics of Bone in Experimental Diabetic rats.

The biomechanics of bone In diabetics and its response to insulin treatment was not clear.We investigated the impace of alloxan-induced diabetes and insulin treatment on the biomechanical characteristics of bones．Sprague-Dawley rats were divided into three groups:

CHRONIC EFFECTS OF EARLY ANGIOTENSIN-CONVERTING ENZYME INHIBITOR (QUINAPRIL) OR ANGIOTENSIN II RECEPTOR BLOCKADE (LOSARTAN) ON HEMODYNAMICS AND REMODELING IN RATS AFTER EXPERIMENTAL MYOCARDIAL INFARCTION

The long-term effects of an angiotensin-converting enzyme (ACE) inhibitor on hemodynamics and remodeling

AN EXPERIMENTAL TRIAL OF ARTEMETHER IN TREATMENT OF PNEUMOCYSTIS CARINII IN IMMUNOSUPPRESSED RATS

An immunosuppressed rat model was establisbed by injecting cortisone acetate 25 mg/rat twice a week for 4 weeks and 12.5mg/rat for another 2 weeks subcutaneously.A development of Pneumocystis carinii pneumonia(PCP) was found at the end of the 6th week in all rats.

Effect of Pressure-Controlled Inverse Ratio Ventilation on WBC,PCT and CRP in Rats Undergoing Radical Gastrectomy and Related Mechanism

Objective:To explore the effect of pressure-controlled inverse ratio ventilation on partial pressure of oxygen(PaO_(2)),partial pressure of carbon dioxide(PaC O2),dynamic lung compliance(Cdyn),white blood cell(WBC),procalcitonin(PCT),C-reactive protein(CRP),and other indicators.Methods:A total of 100 healthy Wistar rats,aged 6-8 weeks and weighing 300-356 g,were used to establish gastric cancer models with 1-methyl-3-nitro-1-nitrosoguanidine(MNNG).60 gastric cancer rats were randomly divided into three groups and treated with laparoscopic radical gastrectomy after anesthesia.Rats in the VCV group were given volume control ventilation,rats in the PCV group were given pressure control ventilation,and rats in the PC-IRV group were given pressure control inverse ratio ventilation.The levels of WBC,PCT and CRP in the three groups were observed and compared on the 1st,3rd and 5th day after operation.Results:There were significant differences in PaO_(2)levels at 15 min after ventilation mode change and at the time of pneumoperitoneum closure,and in PaCO_(2)and Cdyn levels at 10 min after induction and at 15 min after ventilation mode change among the three groups(P<0.05).The PaO_(2)levels in PC-IRV group were higher than those in VCV group and PCV group at 15 min after the ventilation mode was changed and at the time of closing the pneumoperitoneum.The level of PaCO_(2)in PC-IRV group was lower than that in VCV group and PCV group 10 min after induction and 15 min after ventilation mode change,and the level of PaCO_(2)in PC-IRV group was lower than that in VCV group when pneumoperitoneum was turned off.The level of Cdyn in PC-IRV group was higher than that in VCV group and PCV group at 10 min after induction and 15 min after ventilation mode change.The level of Cdyn in PC-IRV group was higher than that in PCV group at the time of closing pneumoperitoneum.The Cdyn level in PCV group was lower than that in VCV group at 15 min after ventilation mode change.There were significant differences in WBC levels among the three groups on the 1st day after operation(P<0.05),and there were no significant differences in PCT and CRP levels among the three groups on the 1st day after operation(P>0.05).There were significant differences in WBC,PCT and CRP levels in the three groups of rats on the 3rd and 5th days after surgery(P<0.05).Except for PCT and CRP on the 1st day after operation,the WBC on the 1st,3rd and 5th days after operation,PCT and CRP on the 3rd and 5th days after operation in PCV group and PC-IRV group were lower than those in VCV group(P<0.05).Except for WBC,PCT and CRP on 1st day after operation,the levels of WBC,PCT and CRP on the 3rd and 5th days after operation in PC-IRV group were lower than those in PCV group(P<0.05).Conclusion:During laparoscopic radical gastrectomy for gastric cancer,pressure-controlled inverse ratio ventilation can help rats maintain good respiratory status and lung compliance during the operation,which can effectively alleviate the postoperative inflammatory response.

Study on the Correlation between IL-6,SAA,CRP and Exertional Heat Stroke in Rats

Objective:To investigate the correlation between interleukin-6(IL-6),serum amyloid A(SAA),C-reactive protein(CRP)and exertional heat stroke in rats.Methods:A total of 90 adult Sprague-Dawley rats without specific pathogens were randomly divided into three groups:control group,classical group and exertion group,with 30 rats in each group.The control group was maintained at a temperature of(26±1)℃and humidity of(60±5)%,while the classical and exertion groups were exposed to a temperature of(40±0.5)℃and humidity of(70±5)%.Additionally,the exertion group underwent treadmill running under these conditions.The levels of IL-6,SAA,and CRP in the three groups were assessed and compared at various time points:before modeling,immediately after onset,and at 2,4,and 8 h after onset.Furthermore,Spearman correlation analysis was employed to examine.Results:The findings from the repeated measures analysis of variance indicated significant variations in the levels of IL-6,SAA,and CRP across the three groups(P<0.05).Both the classic and exertion groups exhibited higher levels of IL-6,SAA,and CRP compared to the control group,with the exertion group demonstrating even higher levels than the classic group(P<0.05).Additionally,Spearman correlation analysis revealed a positive correlation between the onset of heat stroke and the levels of IL-6,SAA,and CRP at the onset,as well as at 2,4,and 8 h post-onset(P<0.05).Furthermore,the types of heat stroke were found to be positively correlated with the levels of IL-6,SAA and CRP(P<0.05).Conclusion:The initiation and nature of heat stroke in rats are significantly associated with the concentrations of IL-6,SAA,and CRP.The concentrations of IL-6,SAA,and CRP within 8 h of onset can reliably forecast the occurrence of heat stroke in rats,serving as a basis for distinguishing classical heat stroke from exertional heat stroke.Nevertheless,the predictive and differentiating efficacy may diminish as the onset time prolongs.

Experimental study of glucose-lowering effects on electroacupuncture at back-shu points in rats

Objective To observe the effects of electroacupuncture(EA)at different acupoints for fasting blood glucose(FBG)and oral glucose tolerance in type 2 diabetes rats so as to verify the glucose-lowering effects of EA.Methods Total100 SD male rats were used as experimental objects。

INVESTIGATION ON EFFECT OF DRUG DOSING REGIMENTS ON DRUG DELIVERY IN SOLID TUMOR VIA LUMPED PARAMETER MODELING AND ANIMAL EXPERIMENTS

This work aims to investigate the effects of dosing regiments on drug delivery in solid tumors and to validate them with experiments on rats. The lumped parameter models of pharmacokinetics and of drug delivery in tumor were developed to simulate time courses of average drug concentration （Ct） of tumor interstitium in two types of dosing regiments （i.e., single-shot and triple-shot ones）. The two regiments were performed via antitumor drug, hydroxycamptothecin （HCPT）, on rats, to measure the drug concentration in the tumor. The simulations of the drug concentration in the tumor of the two dosing regiments were conducted and compared with the experimental data on rats. The coefficients in the models were investigated. It is concluded that the triple-shot method is more effective than that of single-shot injection. The present lumped-parameter model is quantitatively competent for drug delivery in solid tumor.