Objective To investigate the influences of anesthetic meltod on setting up the animal model of orthotopic liver transplantation(OLT) in rats, in order to properly select the anesthetic method. Methods OLT was performed by modified two-cuff technique in Sprague Dawley rats. 100 rats were randomly divided into 5 groups: control group, ether group, ketamine group, chloral hydrate group and pentobarbital group. The anesthetic process, the influences of anesthetic method on physiologic parameters and hepatic function, and the mortality during anhepatic phase of each group were observed. Results The anesthetic process of each anesthetic method was different,ether anesthesia had fewer impacts on physiologic parameters, and the pentobarbital had great hepatotoxicity. High mortality happened in ketamine group, chloral hydrate group and pentobarbital group during anhepatic phase, while no animal died in ether group. Conclusions Each anesthetic method has significant influences on the OLT rat. Ether anesthesia has fewer effects on physiologic parameters and liver function, and has low mortality during anhepatic phase of OLT rats.So it is the best anesthetic method in setting up the animal model of OLT in rats.

Objective To evaluate the correlation between diffusion tensor imaging(DTI)measurements,fiber tracking(FT)and the clinical symptoms in patients with cervical spondylotic myelopathy.Methods According to the Japanese orthopaedics association score(JOA),104 patients with cervical spondylopathy were divided into 4 groups:mild in 31 patients with 13-16 scores,moderate in 27 with 9-12 scores,severe in 25 with 5-8 scores,and serious in 21 with 0-4 scores.According to the lesion signal characters,all patients were divided into 3 groups:Group A with normal signal in both T1 WI and T2WI in 33 patients,Group B with normal signal in T1WI but high signal in T2WI in 30 patients,and Group C with low signal in T1 WI and high signal in T2WI in 41 patients.Apparent diffusion coefficient (ADC),fractional anisotropy(FA),λ1,λ2,λ3 were measured in the spinal cord at the serious pressed section,and fiber tractography was performed.The Spearman correlation analyses was used to correlate each of the DTI measurement with JOA score.Group difference was tested with one-way ANOVA method.Results High quality of DTI was acquired in all patients.The FA values in the mild,moderate,severe,and serious groups were respectively 0.69 ±0.13,0.58 ±0.03,0.46 ±0.08,and 0.37 ±0.11 and significant difference was found in different groups(F =100.59,P ＜ 0.05)and positively correlated with JOA scores (r =0.883,P ＜ 0.05).There was no statistical significance between JOA scores and ADC,λ1,λ2,λ3(r=0.232,0.217,0.113,0.127,P ＞0.05).The FA values in group A,B,and C were respectively 0.67 ±0.33,0.51 ±0.21,0.38 ±0.03,and significant difference was found among different groups(F =50.05,P ＜ 0.05).Decrease of JOA score and high signal in T2 companied with decrease of FA value.Decrease of FA values was found associated with increase of fiber bundle damage.The ADC,λ2,λ3 but not λ1 were significantly different among the JOA groups and the group A,B,and C.Conclusions The FA values are positively correlated with clinical symptoms.Decrease of FA values is found associated with increase of fiber bundle damage.DTI can show the severity and extent of damage of spinal cord in patients with cervical spondylotic myelopathy.

Objective:To establish a quality evaluation method for the simultaneous determination of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin in Danqi Xinmaikang boiled powders and pieces.Methods:Quantitative analysis of multi-components was performed to determine contents of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin with Calycosin-7-O-β-D-Glucopyranoside as the reference substance by single-maker (QAMS). The chromatogram conditions were established, with C18 column as solid phase, acetonitrile-water as flowing phase, 268 nm as detecting wavelength, 1.0 ml/min as flowing rate, 30 ℃ as column temperature, and 10 μl as injection volume.Results:The relative correction factor between Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin was 1.14. There was no significant difference of measured values between the external standard method and QAMS ( P>0.05). With Calycosin-7-O-β-D-Glucopyranoside retention time of 1.00, the relative retention time of Lobetyolin was 1.51 and RSD was less than 5%. Conclusion:It is feasible and accurate to evaluate the quality of Danqi Xinmaikang boiled powders and pieces by QAMS.

Background/Aims: To evaluate the causal correlation between complement components and non-viral liver diseases and their potential use as druggable targets. Methods: We conducted Mendelian randomization (MR) to assess the causal role of circulating complements in the risk of non-viral liver diseases. A complement-centric protein interaction network was constructed to explore biological functions and identify potential therapeutic options. Results: In the MR analysis, genetically predicted levels of complement C1q C chain (C1QC) were positively associated with the risk of autoimmune hepatitis (odds ratio 1.125, 95% confidence interval 1.018–1.244), while complement factor H-related protein 5 (CFHR5) was positively associated with the risk of primary sclerosing cholangitis (PSC;1.193, 1.048– 1.357). On the other hand, CFHR1 (0.621, 0.497–0.776) and CFHR2 (0.824, 0.703–0.965) were inversely associated with the risk of alcohol-related cirrhosis. There were also significant inverse associations between C8 gamma chain (C8G) and PSC (0.832, 0.707–0.979), as well as the risk of metabolic dysfunction-associated steatotic liver disease (1.167, 1.036–1.314). Additionally, C1S (0.111, 0.018–0.672), C7 (1.631, 1.190–2.236), and CFHR2 (1.279, 1.059–1.546) were significantly associated with the risk of hepatocellular carcinoma. Proteins from the complement regulatory networks and various liver diseaserelated proteins share common biological processes. Furthermore, potential therapeutic drugs for various liver diseases were identified through drug repurposing based on the complement regulatory network. Conclusions Our study suggests that certain complement components, including C1S, C1QC, CFHR1, CFHR2, CFHR5, C7, and C8G, might play a role in non-viral liver diseases and could be potential targets for drug development.

OBJECTIVETo investigate the effect of mammalian target of rapamycin(mTOR) inhibitor-rapamycin on cognitive function after chronic cerebral ischemia in mice and its molecular mechanism.

METHODSThe chronic cerebral ischemia model was induced by ligation of right common carotid artery (rUCCAO) in 6-week-old ICR mice. The expressions of mTOR, S6K, S6 and corresponding phosphorylated proteins were detected by Western blotting at different time interval (1 h, 3 h, 6 h, 24 h, 3 d, 7 d, 2 w, 4 w, 6 w) after rUCCAO to determine the changes of mTOR signaling pathway. Rapamycin was administrated i.p. at the dose of 3.0 mg/kg 24 h after rUCCAO. Fluoro Jade B staining was used to detect the apoptotic cells. The expressions of Beclin and LC3-Ⅱ were detected by Western blotting to determine the status of autophagy. Morris water maze test and Y maze test were performed to evaluate cognitive functions.

RESULTSThe mTOR signaling pathway was abnormally activated from 6 h to 6 w after rUCCAO in mouse cortex. The activation of mTOR signaling pathway induced by rUCCAO was reversed by administration of rapamycin, and the apoptotic cell number was significantly decreased (146.1±16.3 vs 84.5±9.6,<0.05). Meanwhile, the elevation of Beclin and LC3-Ⅱ protein induced by rUCCAO was reversed by rapamycin administration. Furthermore, compared with vehicle-treated mice, the latent period[(11.1±2.3) s vs (8.1±1.8) s,<0.05] and swimming distance[(672.8±128.5) cm vs (558.2±124.9) cm,<0.05] were significantly decreased and the number of crossing the platform quadrant in Morris water maze increased(2.8±0.9 vs 5.2±0.8,<0.05) in rapamycin-treated mice. Correct response rate in the Y maze was also increased significantly in rapamycin-treated mice[(38.5±9.2)% vs (64.9±7.9)%,<0.05].

CONCLUSIONSInhibiting mTOR pathway by rapamycin reverses the rUCCAO-induced cognitive impairment partly through the suppression of apoptosis and autophagy.

OBJECTIVE: To investigate the efficacy of brain-targeted rapamycin (T-Rap) in treatment of epilepsy in rats. METHODS: Rapamycin nanoparticles targeting brain were prepared. The epilepsy model was induced by injection of pilocarpine in rats. The rats with pilocarpine-induced epilepsy were treated with rapamycin (Rap group) or brain-targeted rapamycin (T-Rap group). Seizure activity was observed by electroencephalography; the effect on mTOR signaling pathway was detected by Western blot; neuronal death and moss fiber sprouting were analyzed by Fluoro-Jade B (FJB) and Timm's staining, respectively. RESULTS: Electroencephalography showed that both preparation of rapamycin significantly reduced the frequency of spontaneous seizures in rats, and the effect of T-Rap was stronger than that of conventional rapamycin (<0.05). Western blot showed that the phosphorylation levels of S6K and S6 in T-Rap group were lower than those in Rap group (all <0.05), indicating that T-Rap had a stronger inhibitory effect on mTOR signaling pathway. FJB staining showed that T-Rap significantly decreased neuronal death, but there was no significant difference as compared with Rap group. Timm's staining showed that both preparations of rapamycin significantly reduced the germination of mossy fibers, while the effect of T-Rap was more pronounced than Rap group (<0.05). The inhibition of body weight gain of T-Rap group was less than that of Rap group (<0.05). CONCLUSIONS T-Rap has a better therapeutic effect on epilepsy than conventional rapamycin with a less adverse effects in rats.
Animals ; Brain ; drug effects ; Disease Models, Animal ; Epilepsy ; chemically induced ; drug therapy ; Neurons ; drug effects ; Pilocarpine ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Sirolimus ; pharmacology ; therapeutic use ; Treatment Outcome