Application of a novel artificial perfusate based on oxygen-carrying nanoparticles in normothermic machine perfusion for porcine liver preservation after cardiac death / 浙江大学学报·医学版

OBJECTIVE@#To investigate the efficacy of a novel artificial perfusate based on oxygen-carrying perfluoronaphthalene-albumin nanoparticles in normothermic machine perfusion (NMP) for preservation of porcine liver donation after cardiac death.@*METHODS@#Artificial perfusate with perfluoronaphthalene-albumin nanoparticles was prepared at 5% albumin (w/v) and its oxygen carrying capacity was calculated. The livers of 16 Landrace pigs were isolated after 1 h of warm ischemia, and then they were divided into 4 groups and preserved continuously for 24 h with different preservation methods: cold preservation with UW solution (SCS group), NMP preservation by whole blood (blood NMP group), NMP preservation by artificial perfusate without nanoparticles (non-nanoparticles NMP group) and NMP preservation by artificial perfusate containing nanoparticles (nanoparticles NMP group). Hemodynamics, tissue metabolism, biochemical indices of perfusate and bile were monitored every 4 h after the beginning of NMP. Liver tissue samples were collected for histological examination (HE and TUNEL staining) before preservation, 12 h and 24 h after preservation.@*RESULTS@#The oxygen carrying capacity of nanoparticles in 100 mL artificial perfusate was 6.94 μL/mmHg (1 mmHg=0.133 kPa). The hepatic artery and portal vein resistance of nanoparticles NMP group and blood NMP group remained stable during perfusion, and the vascular resistance of nanoparticles NMP group was lower than that of blood NMP group. The concentration of lactic acid in the perfusate decreased to the normal range within 8 h in both nanoparticles NMP group and blood NMP group. There were no significant differences in accumulated bile production, alanine aminotransferase and aspartate aminotransferase in perfusate between nanoparticles NMP group and blood NMP group (all P>0.05). After 24 h perfusion, the histological Suzuki score in blood NMP group and nanoparticles NMP group was lower than that in SCS group and non-nanoparticles NMP group (all P<0.05), and the quantities of TUNEL staining positive cells in blood NMP group and non-nanoparticles NMP group was higher than those in nanoparticles NMP group and SCS group 12 h and 24 h after preservation (all P<0.05).@*CONCLUSION@#Artificial perfusate based on oxygen-carrying nanoparticles can meet the oxygen supply requirements of porcine livers donation after cardiac death during NMP preservation, and it may has superiorities in improving tissue microcirculation and alleviating ischemia-reperfusion injury.

Application prospect of fatty liver in liver transplantation / 中华肝胆外科杂志

Currently liver transplantation is the most effective treatment options for patients with end-stage liver disease, but the lack of donor livers limits the development of liver transplantation. With the increasing usage of marginal donor liver such as fatty donor liver, therefore how to make marginal donor liver play a better role has become a new research hotspot. Although the use of fatty liver as a donor can effectively alleviate the contradiction between recipient demand and donor organ shortage, the risk of early post-transplant graft dysfunction also increases. As donor of fatty liver, the degree of steatoidosis should be moderate or below as far as possible, which significantly reduces postoperative complications and promote the recovery of recipients. Meanwhile, it is essential to monitor the lipid indicators post the surgery, and drug intervention can be added if necessary, which may improve the function of the graft.

Prediction of potential drug interactions of apigenin based on molecular docking and in vitro inhibition experiments / 中国中药杂志

The purpose of this study was to investigate the effect of apigenin on UGT1 A1 enzyme activity and to predict the potential drug-drug interaction of apigenin in clinical use. First,on the basis of previous experiments,the binding targets and binding strength of apigenin to UGT1 A1 enzyme were predicted by computer molecular docking method. Then the inhibitory effect of apigenin on UGT1 A1 enzyme was evaluated by in vitro human liver microsomal incubation system. Molecular docking results showed that apigenin was docked into the active region of UGT1 A1 enzyme protein F,consistent with the active region of bilirubin docking,with moderate affinity. Apigenin flavone mother nucleus mainly interacted with amino acid residues ILE343 and VAL345 to form hydrophobic binding Pi-Alkyl. At the same time,the hydroxyl group on the mother nucleus and the amino acid residue LYS346 formed an additional hydrogen bond,which increased the binding of the molecule to the protein. These results suggested that the flavonoid mother nucleus structure had a special structure binding to the enzyme protein UGT1 A1,and the introduction of hydroxyl groups into the mother nucleus can increase the binding ability. In vitro inhibition experiments showed that apigenin had a moderate inhibitory effect on UGT1 A1 enzyme in a way of competitive inhibition,which was consistent with the results of molecular docking. The results of two experiments showed that apigenin was the substrate of UGT1 A1 enzyme,which could inhibit the activity of UGT1 A1 enzyme competitively,and there was a risk of drug interaction between apigenin and UGT1 A1 enzyme substrate in clinical use.

Comparison of the gallbladder damage caused by microwave ablation and cryoablation in vivo porcine livers.

PURPOSE: To compare the imaging, anatomy, and histopathology of the porcine liver tissue adjacent to the gallbladder, as well as the temperature of the gallbladder wall and the damage degree of gallbladder wall at different times after microwave ablation (MWA) and cryoblation. MATERIALS AND METHODS: Sixteen pigs were randomly divided into MWA group (Group M) and cryoblation group (Group C). The pigs were randomly divided into 8 subgroups according to their execution time, with 2 pigs in every subgroup. The pigs were executed immediately after operation, or at 1‑, 2‑, and 4‑weeks postoperatively according to their assigned subgroup. The imaging and anatomy change of the liver ablation zone and the gallbladder wall were recorded. Histopathological observation was carried out for the damage portion of the gallbladder and the adjacent liver parenchyma. RESULTS: (1) There were no significant statistical differences of the damage degree of the gallbladder between the two groups (P = 0.842). (2) Gallbladder wall edema occurred in Group M immediately after ablation (6/8), of which, 3 cases of gallbladder wall reached full‑thickness damage; overlapping of ice ball and gallbladder wall occurred in Group C (5/8), of which, 4 cases of gallbladder wall reached full‑thickness damage. However, there was neither perforation of gallbladder, biliary fistula, nor liver abscess in all cases. CONCLUSION: Both MWA and cryoablation for liver tissues adjacent to the gallbladder could lead to different damage degrees of the gallbladder wall, but not gallbladder perforation even under the condition of full‑thickness damage.

The research progress on machine perfusion preservation of livers / 中华肝胆外科杂志

Liver transplantation is currently the only treatment option for end-stage liver disease,but it is limited by the lack of high quality donor livers.The ongoing liver donor shortage could be alleviated by the consequent expansion of do-nor criteria to include the use of livers that would once have been discarded as unsuitable,marginal livers.Static cold storage,the current gold standard of organ preservation,is incapable of reversing the ischemic damage these organs have sustained.In hepatic transplantation,the limitations of cold storage are reflected in the significantly higher rates of early al]ograft dysfunction,primary non-function,and local ischemic cholangiopathy,especially seen with older,steatotic and donation after cardiac death (DCD) grafts.The existing cold storage technology has restricted the use of this kind of organs and machine perfusion preservation technology becomes an alternative choice.In this paper,we review the research progress on machine perfusion preservation of livers,and introduce the normothermic machine perfusion.

Role of ATP sensitive potassium channel in protective effects of post-conditioning with hydrogen sulfide against hepatic ischemia-reperfusion injury in rats / 中华麻醉学杂志

ObjectiveTo investigate the role of ATP sensitive potassium (KATP ) channel in protective effects of post-conditioning with hydrogen sulfide against hepatic ischemia-reperfusion (I/R) injury in rats.Methods Thirty adult male SD rats weighing 220-250 g were randomly divided into 5 groups ( n =6 each):sham operation gorup (group S) ; hepatic I/R group; NaHS group; glibenclamide + NaHS group (group G) and 5-hydroxydecanoate (5-HD) + NaHS group (group 5-HD).Hepatic ischemia was produced by 60 min occlusion of left and middle lobe blood flow (about 70％ of the whole liver blood flow) followed by 6 h reperfusion in groups I/R,NaHS,G and 5-HD.NaH S 28 μmol/kg was administered intraperitoneally (IP) at 5 min before reperfusion in groups NaHS,G and 5-HD.Glibenclamide 6 mg/kg and 5-HD 10 mg/kg (KATP channel blockers) were administered IP at 5 min before IP NaHS in groups G and 5-HD respectively.Blood samples were collected at the end of 6 h of reperfusion for measurement of ALT and AST activities and liver specimens were obtained for microscopic examination and measurement of TNF-α content and MPO activity.ResultsLiver I/R significantly increased ALT,AST and MPO activities and TNF-α concentration in group I/R as compared with group S.NaHS administered at 5 min before reperfusion significantly attenuated hepatic I/R injury.Glibenclamide and 5-HD administered before NaHS could block the protective effect of NaHS against hepatic I/R injury.ConclusionHydrogen sulfide postconditioning can protect the liver from I/R injury by activating KATP channel.

Study on Insulin Resistance of Preventing and Treating Experimental Diabetic Rats by Powder of Detoxicating and Activating the Collaterals and Regulating Livers / 中国中医药信息杂志

Objective To research the effects of powder of detoxicating and activating the collaterals and regulating livers on diabetic rats models caused by high fat feed and streptozotocin, to provide scientific basis for clinical dosage. Methods Wistar rats were feeded by high fat feed for one month and forbidden to eat for twelve hours, then injected through abdomen with 30 mg/kg of 1.2% streptozotocin. Blank group was injected with the same amount of lemon buffer solution. One week later, sugar tolenrance test was conducted. Rats affected by diabetics were divided into model group, dimethyldiguanide group, Pyrrole group and group of detoxicating and activating the collaterals and regulating livers. Treatment group was perfused into stomach with different medicine by the dosage of 0.5, 5, 0.5 g/(kg?d) respectively. Blank group and model group were perfused with the same amount of physiological saline. Two months after the dosage, indexes such as saccharogenic hemoglobian and blood lipid were detected, and the sensitive indexes of insulin was calculated. Results In the twelfth week, every index in treatment group by powder of detoxicating and activating the collaterals and regulating livers was much lower than model group (P

Laparoscopic fenestration of hepatic and renal cysts and polycystic livers and kindneys / 中国微创外科杂志

Ojective To study the method of laparoscopic fenestration and its effect for the treatment of hepatic and renal cysts and polycystic livers and kidneys. Methods Laparoscopic fenestration of the cysts was performed Results All of the 15 cases were operated on successfully and recovered smoothly. Conclusions laparoscopic fenestration of hepatic and renal cysts and polycystic livers and kidneys has all the advantages of minimally invasive surgery. Relapsed cysts can be also treated by laparoscopic fenestration.

Oxidative Damage Effect of Particulate Matter (PM_(2.5)) on the Livers, Spleens, Kidneys of Rats / 环境与健康杂志

Objective To investigate the effects of airborne fine particulate matter(PM2.5) on anti-oxidative enzymes activities and lipid peroxidation levels in livers, spleens, and kidneys of rats. Methods 32 male Wistar rats were randomly divided into PM2.5 exposure groups of different concentration (1.5, 7.5, 37.5 mg/kg), exposed by tracheoperfusion and control group treated with physiological saline. Rats were killed 24 h after treatment, and the levels of thiobarbituric acid reactive substance (TBARS), glutathione(GSH) and activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) were determined. Results PM2.5 exposure caused significantly decrease of SOD, CAT, GSH-Px, SOD/TBARS in livers and kidneys in a dose-dependent manner compared with control group (P

Expressions of c-fos and c-myc genes during 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB)-induced rat hepatocarcinoma

We investigated the expression of the growth-related nuclear proto-oncogenes, c-fos and c-myc, in early preneoplastic regions and tumor nodules of 3'-MeDAB induced rat hepatocarcinoma. To amplify the levels of these transcripts, we gave cycloheximide (100 mg/kg B.W. i.p.) to each group of rats. The elevated levels of the 2.2 kb c-fos and 2.4 kb c-myc transcripts appeared as early as the 2nd week after feeding on the 3'-MeDAB diet and lasted through the 4th; 6th weeks and tumor. Southern blot analysis indicated that gross amplification or rearrangements were not observed in DNA of the preneoplastic livers and hepatoma nodules. We also measured the rate of the incorporation of [3H] thymidine into hepatic DNA in order to monitor the rate of cell proliferation occurring at the early preneoplastic periods. We have found that the rate of [3H] thymidine incorporation corresponds to the elevated levels of c-fos and c-myc transcripts in the precancerous stages. This finding suggests that the elevated expressions of c-fos and c-myc may result from the continuous cell proliferative stimuli generated in the carcinogen altered cells, which is essential to the initiation and promotion of chemical hepatocarcinogenesis.

Induction of hepatic drug metabolizing enzymes by DL-methionine in rats.

A single intraperitoneal injection of DL-methionine (500 mg/kg body wt.) to adult male Wistar rats was shown to significantly induce all the components of the hepatic microsomal mixed function oxidase system such as NADPH cytochrome C reductase activity, cytochromes P-450 and b5, as well as activities of drug metabolizing enzymes such as aminopyrine demethylase and uridine 5'-diphosphate-glucuronosyltransferase. Combined administration of nicotinamide (250 mg/kg body wt.) and DL-methionine (500 mg/kg body wt.) was shown to bring about an additional increase (25-30%) in the activities of these enzymes as compared to their induction on independent administration of the two endobiotics. In rats bearing Yoshida sarcoma (ascites) tumour as well as in normal rats injected with serum from tumour bearing animals, the decreased activities of hepatic mixed function oxidases could be restored to their normal levels by administration of DLmethionine (500 mg/kg body wt.) to these rats. Whereas actinomycin D (1 mg/kg body wt.) had no effect on the increased incorporation of [14C] labelled leucine into microsomal proteins following administration of nicotinamide, the enhanced incorporation of the label following DL-methionine administration was completely inhibited by the same dose of actinomycin D. Administration of cycloheximide (0·5 mg/kg body wt.) to rats could completely inhibit the increased incorporation of [14C] leucine into hepatic microsomal proteins following independent administration of nicotinamide and DL-methionine. Similar inhibitory pattern with actinomycin D and cycloheximide was also demonstrated in case of induction of NADPH cytochrome C reductase activity by both these endobiotics.