Detection and differentiation of early hepatocellular carcinoma from cirrhosis using CT perfusion in a rat liver model

BACKGROUND： Functional imaging such as CT perfusion can detect morphological and hemodynamic changes in he- patocellular carcinoma （HCC）. Pre-carcinoma and early HCC nodules are difficult to differentiate by observing only their hemodynamics changes. The present study aimed to investi- gate hemodynamic parameters and evaluate their differential diagnostic cut-off between pre-carcinoma and early HCC nodules using CT perfusion and receiver operating characteristic （ROC） curves. METHODS： Male Wistar rats were randomly divided into control （n=20） and experimental （n=70） groups. Diethylnitrosamine （DEN） was used to induce pre-carcinoma and early HCC nodules in the experimental group. Perfusion scanning was carried out on all survival rats discontinuously from 8 to 16 weeks. Hepatic portal perfusion （HPP）, hepatic arterial fraction （HAF）, hepatic arterial perfusion （HAP）, hepatic blood volume （HBV）, hepatic blood flow （HBF）, mean transit time （MTT） and permeability of capillary vessel surface （PS） data were provided by mathematical deconvolution model. The perfusion parameters were compared among the three groups of rats （control, pre-carcinoma and early HCC groups） using the Kruskal-Wallis test and analyzed with ROC curves. Histological examination of the liver tissues with hematoxylin and eosin staining was performed after CT scan.RESULTS： For HPP, HAF, HBV, HBF and MTT, there were significant differences among the three groups （P〈0.05）. HAF had the highest areas under the ROC curves： 0.80 （control vs pre-carcinoma groups） and 0.95 （control vs early HCC groups） with corresponding optimal cut-offs of 0.37 and 0.42, respectively. The areas under the ROC curves for HPP was 0.79 （control vs pre-carcinoma groups） and 0.92 （control vs early HCC groups） with corresponding optimal cut-offs of 136.60 mL/min/100 mg and 108.47 mL/min/100 mg, respectively. CONCLUSIONS： CT perfusion combined with ROC curve analysis is a new diagnosis model for distinguishing between pre-carcinoma and early HCC nodules. HAF and HPP are the ideal reference indices.

Extended Generalized Riccati Equation Mapping for Thermal Traveling-Wave Distribution in Biological Tissues through a Bio-Heat Transfer Model with Linear/Quadratic Temperature-Dependent Blood Perfusion

Analytical thermal traveling-wave distribution in biological tissues through a bio-heat transfer (BHT) model with linear/quadratic temperature-dependent blood perfusion is discussed in this paper. Using the extended generalized Riccati equation mapping method, we find analytical traveling wave solutions of the considered BHT equation. All the travelling wave solutions obtained have been used to explicitly investigate the effect of linear and quadratic coefficients of temperature dependence on temperature distribution in tissues. We found that the parameter of the nonlinear superposition formula for Riccati can be used to control the temperature of living tissues. Our results prove that the extended generalized Riccati equation mapping method is a powerful tool for investigating thermal traveling-wave distribution in biological tissues.

A Mathematical Model to Solve Bio-Heat Transfer Problems through a Bio-Heat Transfer Equation with Quadratic Temperature-Dependent Blood Perfusion under a Constant Spatial Heating on Skin Surface

We consider the one-dimensional bio-heat transfer equation with quadratic temperature-dependent blood perfusion, which governs the temperature distribution inside biological tissues. Using an extended mapping method with symbolic computation, we obtain the exact analytical thermal traveling wave solution, which describes the non-uniform temperature distribution inside the bodies. The found exact solution is used to investigate the temperature distribution in the tissues. It is found that the surrounding medium with higher temperature does not necessarily imply that the tissue will quickly (after a short duration of heating process) reach the desired temperature. It is also found that increased perfusion causes a decline in local temperature.

Gradual Clamping Reduced Ischemia-Reperfusion Injury in an Isolated Rat Heart Model

Objectives: We hypothesized that the organisms and their organs or tissues could adapt themselves to the gradual changes of environment for surviving or reducing damage. This study explored whether gradual clamping (GC) could reduce myocardial ischemia-reperfusion (IR) injury in rat heart. Methods: Twelve rats were randomized to IR group and GC group, then the hearts were isolated and perfused with Langendorff apparatus. Before cardioplegia, the perfusion was stopped abruptly in IR group while slowly with 5-minute in GC group. The hearts were subjected to 30-minute ischemia and 60-minute reperfusion. The left ventricular develop pressure (LVDP) and systolic pressure (LVSP), the maximal rate of the increase and decrease of left ventricular pressure (+dp/dtmax, ﹣dp/dtmax) were measured by polygraph system at different time points. The recovery of the variables was expressed as the ratio of these values at individual time point after reperfusion to the baseline respectively. Results: The recovery of LVDP after reperfusion was better than that in IR group (P = 0.034). No significant difference in the recovery of LVSP, +dp/dtmax and ﹣dp/dtmax between groups was observed. Conclusions: Gradual clamping could improve the recovery of LVDP after IR, suggesting that gradual clamping could reduce myocardial IR injury.

Effects of Chuanxiongqin hydrochloride on increasing the fluidity of brain cell membrane and scavenging free radicals in model rats with ischemia/reperfusion injury

BACKGROUND: The fluidity of cell membrane can be affected by various factors. Many experiments have confirmed that the ischemia/reperfusion of organic tissue can increase the contents of free radicals, which lead to high rigidity and low fluidity of cell membrane, and the conditions can be changed by Chuanxiongqin. OBJECTIVE: To observe the effect and mechanism of Chuanxiongqin hydrochloride on the fluidity of brain cell membrane in rat models of ischemia/reperfusion. DESIGN: A completely randomized controlled animal trial. SETTINGS: Institute of Brain Sciences; Department of Physiology, Medical College, Datong University. MATERIALS: Twenty male grade Ⅰ Wistar rats of 170-220 g were randomly divided into model group (n =10) and control group (n =10). Chuanxiongqin hydrochloride (molecular mass was 172.2) was purchased from the National Institute for the Control of Pharmaceutical and Biological Products (batch number: 0817-9803); Spin labelers: 5-doxyl-stearlic acid methylester (5DS), 16-doxyl-stearlic acid methylester (16DS), xanthine, xanthine oxidase (XOD) and 5,5-dimeth-1-pyrroline- N-oxide (DMPO) from Sigma Company; Bruker ESP 300 electron paramagnetic resonance (EPR) spectrometer by Bruker Company (Germany). METHODS: The experiments were carried out in the State Key Laboratory of Natural and Biomimetic Drugs, Peking University from June 2001 to July 2002. In the model group, rats were made into models of cerebral ischemia by 30-minute ligation and 2-hour reperfusion of common carotid arteries; The rats in the control group were not made into models. The order parameter (S) and rotational correlation time (τc) were detected with the ESR spectrometer by means of spin labeling. The greater the S and τc, the smaller the fluidity. Meanwhile, the clearance rate of free radicals was detected with ESR spin trapping. The measurement data were compared using the t test. MAIN OUTCOME MEASURES: The S, τc and clearance rates of O2 · and OH· free radicals were compared between the model group and control group. RESULTS: The S and τc in the model group [0.738 4±0.003 5; (8.472±0.027)×10-10 s/circle] were obviously different from those in the control group [0.683 9±0.008 3; (7.945±0.082)×10-10 s/circle, t =5.731, 5.918, P < 0.05], which suggested that ischemia/reperfusion injury decreased the fluidity of brain cell membrane. After adding Chuanxiongqin hydrochloride, there were no obvious differences between the model group [0.688 5±0.030 5; (7.886±0.341)×10-10 s/circle] and control group (P > 0.05), indicating that Chuanxiongqin hydrochloride could recover the fluidity of brain cell membrane after ischemia/reperfusion injury close to the level in the normal control group. Chuanxiongqin hydrochloride could directly scavenge the O2 · and OH· free radicals, and the maximal clearance rates were 83.92% and 44.99% respectively. CONCLUSION: Chuanxiongqin hydrochloride increases the fluidity of membrane of ischemia-injured brain cell by scavenging both O2 ·and OH· free radicals.

Early changes of hepatic hemodynamics measured by functional CT perfusion in a rabbit model of liver tumor

BACKGROUND:Early detection and treatment of hepatocellular carcinoma is crucial to improving the patients’ survival.The hemodynamic changes caused by tumors can be serially measured using CT perfusion.In this study,we used a CT perfusion technique to demonstrate the changes of hepatic hemodynamics in early tumor growth,as a proof-of-concept study for human early hepatocellular carcinoma.METHODS:VX2 tumors were implanted in the liver of ten New Zealand rabbits.CT perfusion scans were made 1 week(early) and 2 weeks(late) after tumor implantation.Ten normal rabbits served as controls.CT perfusion parameters were obtained at the tumor rim,normal tissue surrounding the tumor,and control liver;the parameters were hepatic blood flow,hepatic blood volume,mean transit time,permeability of capillary vessel surface,hepatic arterial index,hepatic arterial perfusion and hepatic portal perfusion.Microvessel density and vascular endothelial growth factor were correlated.RESULTS:At the tumor rim,compared to the controls,hepatic blood flow,hepatic blood volume,permeability of capillary vessel surface,hepatic arterial index,and hepatic arterial perfusion increased,while mean transit time and hepatic portal perfusion decreased on both early and late scans(P<0.05).Hepatic arterial index increased(135%,P<0.05),combined with a sharp increase in hepatic arterial perfusion(182%,P<0.05) and a marked decrease in hepatic portal perfusion(-76%,P<0.05) at 2 weeks rather than at 1 week(P<0.05).Microvessel density and vascular endothelial growth factor showed significant linear correlations with hepatic blood flow,permeability of capillary vessel surface and hepatic arterial index,but not with hepatic blood volume or mean transit time.CONCLUSION:The CT perfusion technique demonstrated early changes of hepatic hemodynamics in this tumor model as proof-of-concept for early hepatocellular carcinoma detection in humans.

EVALUATION OF CIRRHOTIC LIVER WITH PERFUSION-WEIGHTED MAGNETIC RESONANCE IMAGING:A PRELIMINARY EXPERIMENTAL STUDY IN ANIMAL MODELS WITH HALF-LIVER CIRRHOSIS

Objective To investigate the role of peffusion-weighted magnetic resonance imaging （MRI） in evaluation of cirrhotic fiver. Methods With a 4F catheter, 1% diluted carbon tetrachloride （ 1 ml/kg） was selectively injected into fight or left hepatic artery of 12 dogs fortnightly. The half fiver into which carbon tetrachloride was injected was called as study side （SS）, while the other half fiver without carbon tetrachloride injection was called as study control side （SCS）. Conventional and peffusion-weighted MRI were performed in every 4 weeks. Via a 4F catheter, 5ml gadolinium diethylentriamine pentaaceti acid （Gd-DTPA） dilution was injected into superior mesenteric artery at the 5th scan. The signal intensity-thne curves of SS, SCS, and portal vein were completed in MR workstation. The maximal relative signal increase （ MRSI）, peak time （ tp）, and slope of the curves were measured. Results On conventional MR images, no abnormalities of externality and signal intensity were observed in both SS and SCS of fiver at each stage. The mean tp, MP, SI, and slope of intensity-time curves in normal fiver were 10. 56 seconds, 1.01, and 10. 23 arbitrary unit （au）/s, respectively. Three parameters of curves didn＇t show obvious change in SCS of fiver at every stage. Abnormal perfusion curves occurred in SS of fiver at the 12th week after the 1st injection. The abnormality of perfusion curve in SS was more and more serious as the times of injection increased. The mean tp, IVlRSI, and slope intensity-time curves in SS of fiver were 19.45 seconds, 0. 43, and 3. 60 au/s respectively at the 24th week. Conclusion Perfusion-weighted imaging can potentially provide information about portal peffusion of hepatic parenchyma, and to some degree, reflect the severity of cirrhosis.

Quantitative Mitochondrial Proteomics Study on Protective Mechanism of Grape Seed Proanthocyanidin Extracts Against Ischemia/Reperfusion Heart Injury in Rat

Cardiac ischemia/reperfusion（I/R） injury is a critical condition,often associated with high morbidity and mortality.The cardioprotective effect of grape seed proanthocyanidin extracts（GSPE） against oxidant injury during I/R has been described in previous studies.However,the underlying molecular mechanisms have not been fully elucidated.This study investigated the effect of GSPE on reperfusion arrhythmias especially ventricular tachycardia（VT） and ventricular fibrillation（VF）,the lactic acid accumulation and the ultrastructure of ischemic cardiomyocytes as well as the global changes of mitochondria proteins in in vivo rat heart model against I/R injury.GSPE significantly reduced the incidence of VF and VT,lessened the lactic acid accumulation and attenuated the ultrastructure damage.Twenty differential proteins related to cardiac protection were revealed by isobaric tag for relative and absolute quantitation（iTRAQ） profiling.These proteins were mainly involved in energy metabolism.Besides,monoamine oxidase A（MAOA） was also identified.The differential expression of several proteins was validated by Western blot.Our study offered important information on the mechanism of GSPE treatment in ischemic heart disease.

Animal models of ex vivo lung perfusion as a platform for transplantation research

Ex vivo lung perfusion(EVLP) is a powerful experimental model for isolated lung research. EVLP allows for the lungs to be manipulated and characterized in an external environment so that the effect of specific ventilation/perfusion variables can be studied independent of other confounding physiologic contributions. At the same time,EVLP allows for normal organ level function and real-time monitoring of pulmonary physiology and mechanics. As a result,this technique provides uniqueadvantages over in vivo and in vitro models. Small and large animal models of EVLP have been developed and each of these models has their strengths and weaknesses. In this manuscript,we provide insight into the relative strengths of each model and describe how the development of advanced EVLP protocols is leading to a novel experimental platform that can be used to answer critical questions in pulmonary physiology and transplant medicine.

Effects of low molecular heparin combined with Roy adaptation model on hypercoagulable state, endothelial function and placental blood perfusion in patients with preeclampsia

Objective: To investigate the effects of low molecular heparin combined with Roy adaptation model on hypercoagulable state, endothelial function and placental blood perfusion in patients with preeclampsia. Methods: A total of 71 patients with preeclampsia who were treated in Zigong Third People's Hospital between December 2014 and February 2017 were retrospectively analyzed and divided into the control group (n=38) who accepted conventional low molecular heparin therapy and the study group (n=33) who accepted low molecular heparin combined with Roy adaptation model therapy. The differences in hypercoagulable state, endothelial function and placental blood perfusion were compared between the two groups before intervention and after 8 weeks of intervention. Results: Before intervention, there was no statistically significant difference in the hypercoagulable state, endothelial function and placental blood perfusion between the two groups of patients. After 8 weeks of intervention, peripheral blood coagulation indexes TT and AT-Ⅲ levels of study group were higher than those of control group while D-D level was lower than that of control group;serum endothelial function index NO content was higher than that of control group while ET-1 content was lower than that of control group;ultrasonic placental blood perfusion parameters FI, VI and VFI levels were higher than those of control group. Conclusion: Low molecular heparin combined with Roy adaptation model intervention could further reduce the hypercoagulable state, decrease the vascular endothelial injury, and eventually increase the placental blood perfusion in patients with preeclampsia.

Quantification of angiogenesis by CT perfusion imaging in liver tumor of rabbit

BACKGROUND:Tumor angiogenesis is essential for primary and metastatic tumor growth.Computed tomography perfusion(CTP)is a new imaging method,made possible by the recent development of fast CT scanners and improved data analysis techniques,which allows measurement of the physiologic and hemodynamic properties of tissue vasculature.This study aimed to evaluate CTP in the quantification of angiogenesis and to assess the relationship between tissue perfusion parameters and microvascular density(MVD)and vascular endothelial growth factor(VEGF),attempting to detect the physiologic properties of angiogenesis.METHODS:Sixteen rabbits with VX2 liver tumors underwent multi-slice CT perfusion(MSCTP)on day 14 after tumor inoculation.CTP parameters included hepatic blood flow(HBF),hepatic blood volume(HBV),mean transit time(MTT),permeability of capillary vessel surface(PS),hepatic artery index(HAI),hepatic artery perfusion(HAP),and hepatic portal perfusion(HPP).The border of the tumor was stained with CD34 and VEGF immunohistochemical stains,and MVD was measured by anti-CD34.Then,CTP parameters were determined whether they were correlated with MVD and VEGF using Pearson’s correlation coefficient.RESULTS:The positive expression of MVD was different in the center and border of the tumor(P【0.01).There was a positive correlation between MVD and VEGF in the border(P【0.05).As more VEGF was expressed,the number of microvessels increased.Correlation analyses were also made between the perfusion parameters and MVD and VEGF in the border of the tumor.HBF,PS,HAI,and HAP values were positively correlated with MVD and VEGF(P【0.05),HPP was negatively correlated with MVD and VEGF(P【0.01),and HBV and MTT values were not correlated with MVD and VEGF(P】0.05).CONCLUSIONS:Significant correlations were found between perfusion parameters and MVD and VEGF.Therefore,MSCTP can be used to evaluate tumor angiogenesis in vivo.

Assessment of hemodynamics in precancerous lesion of hepatocellular carcinoma:Evaluation with MR perfusion

AIM: To investigate the hemodynamic changes in a precancerous lesion model of hepatocellular carcinoma (HCC). METHODS: Hemodynamic changes in 18 Wistar rats were studied with non-invasive magnetic resonance (MR) perfusion. The changes induced by diethylnitrosamine (DEN) developed into liver nodular lesions due to hepatic cirrhosis during the progression of carcinogenesis. The MR perfusion data [positive enhancement integral (PEI)] were compared between the nodular lesions corresponding well with MR images and pathology and their surrounding hepatic parenchyma. RESULTS: A total of 46 nodules were located by MR imaging and autopsy, including 22 dysplastic nodules (DN), 9 regenerative nodules (RN), 10 early HCCs and 5 overt HCCs. Among the 22 DNs, 6 were low-grade DN (lGDN) and 16 were high-grade DN (HGDN). The average PEI of RN, DN, early and overt HCC was 205.67 ± 31.17, 161.94 ± 20.74, 226.09 ± 34.83, 491.86 ± 44.61 respectively, and their liver parenchyma nearby was 204.84 ± 70.19. Comparison of the blood perfusion index between each RN and its surrounding hepatic parenchyma showed no statistically significant difference (P = 0.06). There were significant differences in DN (P = 0.02). During the late hepatic arterial phase, the perfusion curve in DN declined. DN had an iso-signal intensity at the early hepatic arterial phase and a low signal intensity at the portal venous phase. Of the 10early HCCs, 4 demonstrated less blood perfusion and 6 displayed minimally increased blood flow compared to the surrounding parenchyma. Five HCCs showed significantly increased blood supply compared to the surrounding parenchyma (P = 0.02). CONCLUSION: Non-invasive MR perfusion can detect changes in blood supply of precancerous lesions.

Desferrioxamine in warm reperfusion media decreases liver injury aggravated by cold storage

AIM:To evaluate whether desferrioxamine decreases ischemia and perfusion injury aggravated by cold storage(CS)in a rat liver perfusion model. METHODS:Isolated rat livers were kept in CS in University of Wisconsin Solution for 20 h at 4℃,then exposed to 25 min of warm ischemia(WI)at 37℃ followed by 2 h of warm perfusion(WP)at 37℃with oxygenated(95%oxygen and 5%carbon dioxide) Krebs-Henseleit buffer.Desferrioxamine(DFO),an iron chelator,was added at different stages of storage,ischemia and perfusion:in CS only,in WI only,in WP only, in WI and perfusion,or in all stages.Effluent samples were collected after CS and after WI.Perfusate samples and bile were collected every 30 min(0,0.5,1,1.5 and 2 h)during liver perfusion.Cellular injury was assessed by the determination of lactate dehydrogenase(LDH) and aspartate aminotransferase(AST)in the effluent and perfusate samples.Total iron was analysed in the perfusate samples.After WP,the liver was collected for the determination of liver swelling(wet to dry ratio) and liver morphological examination(hematoxylin and eosin staining). RESULTS:Increased CS time caused increased liver dysfunction during WP.After 2 h of WP,liver injury was indicated by increased release of AST(0.5 h CS:9.4± 2.2 U/g liver vs 20 h CS:45.9±10.8 U/g liver,P<0.05) and LDH(0.5 h CS:59±14 U/g liver vs 20 h CS:297 ±71 U/g liver,P<0.05).There was an associated increase in iron release into the perfusate(0.5 h CS:0.11 ±0.03μmoL/g liver vs 20 h CS:0.58±0.10μmoL/g liver,P<0.05)and reduction in bile flow(0.5 h CS: 194±12μL/g vs 20 h CS:71±8μL/g liver,P<0.05). When DFO was added during WI and WP following 20 h of CS,release of iron into the perfusate was de- creased(DFO absent 0.58±0.10μmoL/g liver vs DFO present 0.31±0.06μmoL/g liver,P<0.05),and liver function substantially improved with decreased release of AST(DFO absent 45.9±10.8 U/g liver vs DFO present 8.1±0.9 U/g liver,P<0.05)and LDH(DFO absent 297±71 U/g liver vs DFO present 56±7 U/g liver,P<0.05),and increased bile flow(DFO absent 71±8μL/g liver vs DFO present 237±36μL/g liver, P<0.05).DFO was also shown to improve liver morphology after WP.Cellular injury(the release of LDH and AST)was significantly reduced with the addition of DFO in CS medium but to a lesser extent compared to the addition of DFO in WP or WI and perfusion.There was no effect on liver swelling or bile flow when DFO was only added to the CS medium. CONCLUSION:DFO added during WI and perfusion decreased liver perfusion injury aggravated by extended CS.

Qualitative and Quantitative Perfusion Parameters Determined by 3D Single-Shot GRASE ASL MR Imaging

Rationale and Objectives: A particular arterial spin (ASL) labeling technique, called 3D-single-shot GRASE ASL is discussed with respect to the ability and limits of quantifying perfusion parameters. Materials and Methods: The technique enables the acquisition of perfusion weighted signal at multiple delay times (TI) in one scan. The readout part is a gradient and spin-echo combination (GRASE) that uses switched gradient rephrasing of signals to produce several times as many signals as turbo-spin-echo, which translates into faster imaging time and higher signal-to-noise ratio (SNR) per imaging time. The technique provides the possibility for model based quantification of cerebral blood flow and the determination of the bolus arrival information without use of contrast agent and thus the characterization and determina-tion of regions that are supported by collaterals. Results: Whereas for a quantification of the permeability using ASL the SNR is not high enough, at least qualitative permeability maps can be determined, if an optimal homogenous SNR was guaranteed. This was accomplished in brain regions with a high blood supply, typically given in tumors, and by using a correction for coil sensitivity at the highest possible additional scaling. Conclusion: The single-shot 3D GRASE ASL can provide information about the principal blood supply, the transit delay of the blood flow due to a stenosis or collaterals and a qualitative measure of the permeability.

Effects of high-frequency oscillatory ventilation and conventional mechanical ventilation on oxygen metabolism and tissue perfusion in sheep models of acute respiratory distress syndrome

Background High-frequency oscillatory ventilation （HFOV） allows for small tidal volumes at mean airway pressures （mPaw） above that of conventional mechanical ventilation （CMV）,but the effect of HFOV on hemodynamics,oxygen metabolism,and tissue perfusion in acute respiratory distress syndrome （ARDS） remains unclear.We investigated the effects of HFOV and CMV in sheep models with ARDS.Methods After inducing ARDS by repeated lavage,twelve adult sheep were randomly divided into a HFOV or CMV group.After stabilization,standard lung recruitments （40 cmH2O × 40 seconds） were performed.The optimal mPaw or positive end-expiratory pressure was obtained by lung recruitment and decremental positive end-expiratory pressure titration.The animals were then ventilated for 4 hours.The hemodynamics,tissue perfusion （superior mesenteric artery blood flow,pHi,and Pg-aCO2）,oxygen metabolism and respiratory mechanics were examined at baseline before saline lavage,in the ARDS model,after model stabilization,and during hourly mechanical ventilation for up to 4 hours.A two-way repeated measures analysis of variance was applied to evaluate differences between the groups.Results The titrated mPaw was higher and the tidal volumes lower in the HFOV group than the positive end-expiratory pressure in the CMV group.There was no significant difference in hemodynamic parameters between the HFOV and CMV groups.There was no difference in the mean alveolar pressure between the two groups.After lung recruitment,both groups showed an improvement in the oxygenation,oxygen delivery,and DO2.Lactate levels increased in both groups after inducing the ARDS model.Compared with the CMV group,the superior mesenteric artery blood flow and pHi were significantly higher in the HFOV group,but the Pg-aCO2 decreased in the HFOV group.Conclusion Compared with CMV,HFOV with optimal mPaw has no significant side effect on hemodynamics or oxygen metabolism,and increases gastric tissue blood perfusion.

Investigation on the optical scan condition for imaging of multi-slice spiral CT liver perfusion in rats

Background Multi-slice CT liver perfusion has been widely used in experimental studies of hemodynamic changes in liver lesions, and is usually performed as an adjunct to a conventional CT examination because of its high temporal and spatial resolution, simple protocol, good reproducibility, and ability to measure hemodynamic changes of liver tissues at the capillary level. Experimental rat models, especially those of induced liver cancer, are often used in studies of hemodynamic changes in liver cancer. Carcinogenesis in rats has a similar pathological progression and characteristics resembling those in human liver cancer; as a result, rat models are often used as ideal animal models in the study of human liver cancer. However, liver perfusion imaging in rats is difficult to perform, because rats＇ livers are so small that different concentrations, flow rates, and dose of contrast agents during the CT perfusion scanning can influence the quality of liver perfusion images in rats. The purpose of this study, therefore, was to investigate the optimal scan protocol for the imaging of hepatic perfusion using a deconvolution mathematical method in rats by comparing the results of rats in different injection conditions of the contrast agent, including concentration, rate and time. Methods Plain CT scan conditions in eighty 2-month-old male Wistar rats were 5.0 mm slice thickness, 5.0 mm interval, 1.0 pitch, 120 kV tube voltage, 60 mA tube current, 512x512 matrix, and FOV 9.6 cm. Perfusion scanning was carried out with different concentrations of diatrizoate （19%, 38%, 57%, and 76%）, different injection rates （0.3 and 0.5 ml/s）, and different injection times （1, 2-3, 4-5, and 6 seconds）. The above conditions were randomly matched and adjusted to determine the best perfusion scan protocol. Three-phase contrast-enhanced scanning was performed after CT perfusion. Histological examination of the liver tissues with hematoxylin and eosin stains was done after CT scanning. Results When the concentration of the contrast agent was 19% or 38%, no pseudo-color map was created. The viscosity increased when the concentration of the contrast agent was 76%; so it is difficult to inject the contrast agent at such a high concentration. Also no pseudo-color map was generated when the injection time was short （1, 2-3, and 4-5 seconds） or the injection rate was low （0.3 ml/s）. The best perfusion images and perfusion parameters were obtained during 50 seconds scanning. Each rat was given an injection of 57% diatrizoate at 0.5 mils via the tail vein using a high-pressure syringe for 6 seconds. The perfusion parameters included hepatic blood flow （HBF）, hepatic blood volume （HBV）, mean transit time （MTT） of the contrast agent, capillary permeability-surface area product （PS）, hepatic arterial index （HAl）, hepatic artery perfusion （HAP）, and hepatic portal perfusion （HPP）. All these parameters reflected the perfusion status of liver parenchyma in normal rats. Three phases of enhancement were modified according to the time-density curves （TDCs） of the perfusion imaging： hepatic arterial phase （7 seconds）, hepatic portal venous phase （15 seconds）, and a delayed phase （23-31 seconds）. On examination by microscopy, the liver tissues were pathologically normal. Conclusions The appropriate protocol with multi-slice spiral CT liver perfusion reflected normal liver hemodynamics in rats. This study laid a solid foundation for further investigation of the physiological characteristics of liver cancer in a rat model, and was an important supplement to and reference for conventional contrast-enhanced CT scans.

Application of gastric perfusion intralipid for formulating rabbit's hyperlipidemia models

Background The commonly used method so far for formulating hyperlipidemia animal models is to feed the animals with a high fat diet. Owning to influences of animal feeding, dosage and cycle, feeding with a high fat diet has resulted in an unstable quality. It is unknown whether gastric perfusion of intralipid can establish a better hyperlipidemia model in rabbits. Methods Twenty rabbits were randomly selected and subjected to gastric intralipid perfusion for consecutive 2 weeks or to feed with common high cholesterol feed for 4 weeks. The levels of blood serum total cholesterol （TC）, triglyceride （TG）, apolipoprotein A1 （APOA1）, APOB, blood plasma thromboxane B2 （TXB2）, 6-Keto-prostaglandin F1α （6-Keto-PGF1α）, endotheline （ET）, erythrocyte membrane cholesterol （M-Tc）, membrane fluidity （M-Flu）, erythrocyte malondialdehyde （E-MDA）, erythrocyte superoxide dismutase （E-SOD） before and after the experiment. In the end of the experiment, the aortae were removed from all the animals for light and electron microscopic examinations. Results The results showed that in both groups, the levels of blood serum TC, TG, APOB, blood plasma TXB2, ET, erythrocyte M-Flu and E-MDA were significantly increased. Blood serum APOA1, blood plasma 6-Keto-PGF1α and E-SOD were significantly decreased. But, erythrocyte M-TC had no significantly changed. The indexes of the two groups were no obvious difference before and after the experiment. Light and electron microscopic examination showed that there were early changes of early atherosclosis in the two groups of animal, in accordance with the characteristics of experimental animal hyperlipidemia. Conclusion The results suggest that gastric perfusion of intralipid can produce hyperlipidemia in rabbits faster than the commonly used model developed by feeding high cholesterol diet.

Comparison of intraperitoneal and intratesticular ozone therapy for the treatment of testicular ischemia-reperfusion injury in rats

We compare the efficacy of intratesticular ozone therapy with intraperitoneal ozone therapy in an experimental rat model. For this purpose, 24 rats were divided into four groups including sham-operated, torsion/detorsion, torsion/detorsion plus intraperitoneal ozone （O-IP）, and torsion/detorsion plus intratesticular ozone （O-IT）. The O-IP ozone group received a 4 mg kg-1 intraperitoneal injection of ozone, and the O-IT group received the same injection epididymally. At 4 h after detorsion, the rats were sacrificed and orchiectomy materials were assessed histopathologically. Spermatogenesis in the seminiferous tubules and damage to the Sertoli cells were histopathologically evaluated in the testes using the Johnsen scoring system, i-NOS and e-NOS activities in the testis tissue were also evaluated. Torsion-detorsion caused a decreased Johnsen score and increased apoptosis of spermatogonial and Sertoli cells. Ozone injection prevented increases in Johnsen score and i-NOS level, e-NOS level of the O-IP group was significantly lower than that of the O-IP group, and i-NOS level of the O-IT group was significantly lower than that of the O-IP group. Local ozone therapy is more effective than systemic ozone therapy at improving IRI-related testicular torsion. Our study is the first to show that the efficacy of intratesticular implementation of ozone therapy is higher than that of intraperitoneal ozone therapy.

Preliminary exploration of animal models of congenital choledochal cysts

BACKGROUND Various animal models have been used to explore the pathogenesis of choledochal cysts(CCs),but with little convincing results.Current surgical techniques can achieve satisfactory outcomes for treatment of CCs.Consequently,recent studies have focused more on clinical issues rather than basic research.Therefore,we need appropriate animal models to further basic research.AIM To establish an appropriate animal model that may contribute to the investigation of the pathogenesis of CCs.METHODS Eighty-four specific pathogen-free female Sprague-Dawley rats were randomly allocated to a surgical group,sham surgical group,or control group.A rat model of CC was established by partial ligation of the bile duct.The reliability of the model was confirmed by measurements of serum biochemical indices,morpho-logy of common bile ducts of the rats as well as molecular biology experiments in rat and human tissues.RESULTS Dilation classified as mild(diameter,≥1 mm to<3 mm),moderate(≥3 mm to<10 mm),and severe(≥10 mm)was observed in 17,17,and 2 rats in the surgical group,respectively,while no dilation was observed in the control and sham surgical groups.Serum levels of alanine aminotransferase,aspartate aminotrans-ferase,total bilirubin,direct bilirubin,and total bile acids were significantly elevated in the surgical group as compared to the control group 7 d after surgery,while direct bilirubin,total bilirubin,and gamma-glutamyltransferase were further increased 14 d after surgery.Most of the biochemical indices gradually decreased to normal ranges 28 d after surgery.The protein expression trend of signal transducer and activator of transcription 3 in rat model was consistent with the human CC tissues.CONCLUSION The model of partial ligation of the bile duct of juvenile rats could morphologically simulate the cystic or fusiform CC,which may contribute to investigating the pathogenesis of CC.

The bio-active components of the Mongolian medicine Horcha-6 and therapeutic mechanism in the rat migraine model

Background:The active components of Horcha-6 were identified using liquid chromatography with tandem mass spectrometry.Also,we investigated the potential mechanisms that explain why Horcha-6 may be effective in treating migraines through the use of network pharmacology and a rat migraine model.Methods:After identifying the active components of Horcha-6,the corresponding genes of the active components’target were obtained from the Universal Protein database,and a“compound-target-disease”network was constructed using Cytoscape 3.9.0 software.For the in vivo experiments,nitroglycerin was injected intraperitoneally into rats to create a migraine model.Pre-treatment with Horcha-6 was administered orally for 14 days,and rats were subjected to migraine-related behavior tests.RNA sequencing was performed to identify the gene expression regulated by Horcha-6 in the trigeminal nerve.Results:A total of 903 chemical components of Horcha-6 have been collected in the liquid chromatography with tandem mass spectrometry.We discovered 55 of the Horcha-6 bio-active components that were evaluated based on their Percent Human Oral Absorption(≥30%)and DL values(≥0.185)on the traditional Chinese medicine systems pharmacology database.The“compound-target-disease”network contained 163 intersection targets with the migraine state.Gene Ontology analysis indicated that these components significantly regulated the immune response,vascular function,oxidative stress,etc.When Kyoto Encyclopedia of Genes and Genomes enrichment analysis was performed,we observed that most of the target genes were significantly enriched in the inflammation and neuro-related signaling pathway,toll-like receptor signaling pathway,neuroactive ligand-receptor interaction,etc.These predictions were further demonstrated via in vivo animal model experiments.The RNA sequencing results showed that 41 genes were down-regulated(P<0.05)and 86 genes were up-regulated(P<0.05)in the Horcha-6 treated group compared with the untreated group.Those genes were mainly involved in neuromodulation,vascular function,and hormone metabolism.Conclusion:The 55 bio-active components in Horcha-6 regulate inflammation,hormone metabolism,and neurotransmitters and have potential as a therapy to treat migraines.