To investigate the semi-quantitative method for evaluating lipid accumulation in livers, male C57BL/6J mice (CON), HFD mice characterized with the mild fatty liver induced by high-fat diet, and KKAy mice charactered with the moderately severe fatty liver induced by high-caloric diet were used. The lipid accumulation was estimated by the histological examination (HE staining) and the content of hepatic triglyceride, separately. Echo-intensity of two selected regions along the ultrasound transmission direction was recorded using a small animal ultrasonographic system, and the echo-intensity attenuation coefficient was calculated. Correlation between the echo-intensity attenuation coefficient and the content of hepatic triglyceride was analyzed by the Spearman's rank correlation analysis. The results showed that the lipid accumulation in livers increased significantly in both HFD and KKAy mice compared with CON mice and it was more serious in KKAy mice than that in HFD mice. The values of echo-intensity attenuation coefficient were also increased in sequence according to group. These values were positively associated with the content of hepatic triglyceride (r = 0.744, P < 0.01). In conclusion, the echo-intensity attenuation coefficient is a simple, impersonal, and non-invasive method for evaluating the hepatic lipid accumulation. It can be used to research the process and the treatment of fatty liver diseases in mice.

Background Experimental autoimmune uveitis (EAU) is a common animal model of uveitis.Natural killer (NK) cells have been confirmed to be a type of strong inflammation-causing cells,but its role in EAU is still studing.Objective This study was designed to explore the role and mechanism of NK cells in the pathogenesis of EAU.Methods Thirty-six SPF Lewis rats were randomly divided into expeimental control group and EAU 6-,9-,12-,16-,and 21-day groups (6 rats for each group).Rats in EAU group received subcutaneous injection interphotoreceptor retinoid binding protein (IRBP) combining 5 mg/ml tubercle bacillus with complete Freund's adjuvant (CFA) emulsion in foot pads,and then 400 ng pertussis toxin was intraperitoneally injected to extablish EAU models in the EAU 6-,9-,12-,16-,and 21-day group,and normal saline solution combined with CFA and 400 ng pertussis toxin was used in the same way in the experimental control group.The inflammatory response was observed by slit lamp daily after modeling and scored based on Caspi criteria.The eyeballs were extracted in 6,9,12,16 and 21 days after modeling for retinal histopathological examination,Immunofluorescent double-staining was employed to detect and locate the expression of NK cells in the retina.In addition,25 model rats were divided into EAU 0-,3-,6-,9-and 12-day groups,with 5 rats for each group,and eyeballs were extracted to prepare tissue homogenate.The expression of CXCL10 mRNA,and CXCL12 mRNA NK cell chemokines,in the tissue homogenate was assayed by real-time quantitative PCR.The use and care of the rats followed Regulations for the Administration of Affair Concerning Experimental Animal by State Science and Technology Commission.Results No inflammatory sign in ocular anterior segment of the rats was seen in the experimental control group.The expansion of rat iris vessels was found in the EAU 6-day group,and exudes and hypopyon of the anterior chamber occurred in the EAU 9-day group and the inflammation peaked in the EAU 12-day gorup.The rat retinal structure was normal in the experimental control group,and the arrangement disorder of retinal structure,the cell separation in outer nuclear layer and damage of photoreceptors were found under the optical microscope in different degree in various EAU groups,with the most serious change in the EAU 12-day group.Immunofluorescent double staining showed normally arranged nucleus in the experimental control group,and a lot of NK infiltration was seen in the EAU 6-day group and peaked in the EAU 9-day group.The expression level of CXCL10 mRNA in the EAU 9-day group was 34.298 ± 16.689,which was significantly higher than that in the EAU 3-,6-and 12-day group,respectively (1.390 ± 0.660,3.359 ± 2.581,4.711 ±1.387) (all at P＜0.01).No significant differences were found in the relative expression of CXCL12 mRNA among different EAU groups (F=2.851,P＞0.05).Conclusions Retinal NK cell infiltration occurs in the early stage of EAU,and the severity of NK cell infiltration is consistent with the inflammatory process and CXCL10 expression,suggesting NK cells play an important role in the early stage of EAU,and CXCL10 is an important chemokine of NK cells in EAU rats.

The effect of realgar nanoparticles (NPs) on endogenous small molecules in rat kidney was analyzed by mass spectrometry-based metabolomics.The relationship between the changes of metabolites and the nephrotoxicity of realgar NPs was also discussed to provide a basis for the further toxicity study and the clinical application of realgar NPs.SD rats were randomly divided into seven groups,including control group,three doses (40,200,1 000 mg/kg) of relegar and realgar NPs groups,respectly.After 28 days of continuous intragastric administration,all rats were sacrificed and their serum and kidney samples were collected.The toxic effect of realgar NPs on kidney tissues were examined by biochemical analysis and histopathologic examination,which revealed a dosedependent nephrotoxicity induced by realgar NPs.The LC-MS and GC-MS analysis were performed for the subsequent metabolomics study.A series of 32 metabolites were found to be altered significandy in the kindey of realgar NPs treated rats,and might serve as potential nephrotoxicity biomarkers.The results of metabolic pathway analysis indicated that the nephrotoxicity of realgar NPs might be associated with the disorders of the amino acids and phosphatidic acid metabolism.

This study examined the current changes of human ether-a-go-go-related gene (hERG) mutation derived from a LQT2 Chinese family with a highly penetrating phenotype. Mutation was identified and site-directed mutagenesis was performed to induce the mutation in wild-type (WT) hERG. WT hERG and mutated V535M were cloned and transiently expressed in HEK293 cells. At the 48th and 72nd h after transfection, membrane currents were recorded using whole cell patch-clamp procedures. An A>G transition at 1605 resulting in replacement of V535M was identified. Compared to WT, V535M mutation significantly decreased tail currents of hERG. At test potential of -40 mV after depolarizing at +50 mV, tail current densities were 83.35±7.06 pA/pF in WT and 50.38±7.74 pA/pF in V535M respectively (n=20, P<0.01). Gating kinetics of hERG revealed that V (1/2) of steady-state inactivation shifted to negative potential in the mutant (V (1/2,V535M): -61.81±1.7 mV vs. V (1/2, WT): -43.1±0.71 mV). The time constant of recovery from inactivation was markedly prolonged in the mutant compared to WT among test potentials. V535M hERG mutation demonstrated markedly decreased tail current densities, which suggests that V535M is a new loss-of-function mutation of hERG channel responsible for LQT2.

Objective To investigate the expression and clinical significances of miR-215 and runtrelated protein1 (RUNX1) in retinoblastoma (RB),and to study the regulation effect of miR-215 on RUNX1 in the retinoblastoma cell line PMC-RB.Methods The expressions of miR-215 and RUNX1 in the tumor tissue,non tumor tissues adjacent to cancer,human RB cell line FMC-RB and human normal retinal vascular endothelial cell line ATCC of RB patients were detected by quantitative real-time polymerase chain reaction (qRT-PCR).miR-215 mimics (miR-215-mimic),miR-NC,si-RUNX1 and si-NC were transfected into FMC-RB cell line respectively.Cell proliferation,migration and invasion ability were measured respectively,thus detecting the regulation effect of miR-215 on RUNX1.Results The expression of miR-215 in RB tissues was significantly lower than that in non tumor tissues adjacent to cancer,while the mRNA expression of RUNX1 was higher than that in non tumor tissues adjacent to cancer (P ＜ 0.05).The expression of miR-215 in PMC-RB cells was lower than that in ATCC,while the mRNA expression of RUNX1 was higher than that in ATCC (P ＜0.05).The expression of miR-215 and RUNX1 in RB tumor tissues were closely related to the clinicopathological features of optic nerve infiltration,tumor tissue differentiation and lymph node metastasis (P ＜ 0.05).Cell proliferation,migration and invasion in miR-215-mimic group were significantly lower than those in miR-NC group (P ＜ 0.05).In transfected 3' untranslated region (3 'UTR)-Wt cells,the luciferase activity in miR-215-mimic group was lower than that in miR-NC group (P ＜ 0.05);the expression level of RUNX1 protein in transfected miR-215-mimic cells was lower than that in transfected miR-NC cells (P ＜ 0.05).Cell proliferation,migration and invasion in si-RUNX1 group were all lower than those in si-NC group (P ＜ 0.05).There was a negative correlation between mRNA expression level of miR-215 and RUNX1 in RB tumor tissues.Conclusions During the occurrence and development of RB,the down-regulation of miR-215 expression can promote malignant progression of tumor by targeting RUNX1.miR-215 can be used as a biological markers and therapeutic target for RB diagnosis.

Grey system theory was applied in analysis of Electroencephalogram (EEG) to extract features of driving fatigue in this study. Model GM(1,1) was built for EEG collected during simulative driving experiments. At the same time, the data of steering wheel movements and subjective fatigue level were analyzed as reference. The results of experiments reveal that the co-deviation of Model GM(1,1) parameter a and b, cov(a,b), coincides with the standard deviation of steering wheel movements. This indicates that Grey system theory is effective for EEG analysis and the parameters of GM(1,1) can well reflect the change of driving fatigue.
Adult ; Automobile Driving ; psychology ; Computer Simulation ; Electroencephalography ; methods ; Fatigue ; physiopathology ; Humans ; Male ; Models, Theoretical

Objective: To analyze the trends in mortality of liver cancer in Wenzhou City, Zhejiang Province from 2014 to 2022, so as to provide the evidence for improving liver cancer control measures. Methods: Data of liver cancer mortality in Wenzhou City from 2014 to 2022 were collected from Wenzhou Chronic Disease Monitoring Information System. The crude mortality were estimated and standardized by the national population census data in China in 2010, and the trends in mortality of liver cancer were analyzed with average annual percent change (AAPC). Results: There were 22 033 liver cancer deaths from 2014 to 2022, accounting for 18.08% of malignant tumor deaths and ranking the second in malignant tumor deaths. The crude mortality of liver cancer was 30.00/105 and the standardized mortality was 24.32/105, both showing decreasing trends (AAPC=-2.812% and -5.742%, both P<0.05). The standardized mortality of liver cancer were higher in men than in women (36.66/105 vs. 11.21/105, P<0.05), both showing decreasing trends (AAPC=-5.702% and -5.521%, both P<0.05). The crude mortality of liver cancer appeared a tendency towards a rise with age (P<0.05), with the highest crude mortality in the group aged 80 to 84 years, reaching 145.12/105. The crude mortality of liver cancer showed a tendency towards a decline among residents aged under 15 years, 15 to 44 years, 45 to 64 years and 65 years and above (AAPC=-20.311%, -6.569%, -7.408% and -3.177%, all P<0.05). Conclusions The mortality of liver cancer showed a tendency towards a decline in Wenzhou City from 2014 to 2022. Men and the elderly were high-risk groups for liver cancer deaths, and prevention should be strengthened based on risk factors.

Objective To build a multi‐level hierarchical structure model of the influencing factors for hierarchical medical system ,to identify the role relationship between all the factors and transmission pathways ,and to recommend on developing China′s hierarchical medical system . Methods Thirty influencing factors were identified in a screening based on literature review for the hierarchical medical system .On such basis ,16 influencing factors were identified by three health policy experts ,which affect operations of the current system .Interpretative structural modeling was called into play in the end to analyze the hierarchy relationship between various influencing factors and the conduction loops .Results There exist among the 16 factors a 3‐level hierarchical structureand two conduction loops .The factor directly limiting the hierarchical medical system is two‐way referral,and most internal core drivers arehuman resources development and governance mechanism.By means of self‐growth and external constraints ,they exert their influence on the operation of hierarchical medical system .Conclusions There are interactive hierarchical effects among the factors ,merging into three node elements of functional role,inter‐entity relationshipand patient participation.

OBJECTIVETo explore the correlation between metabolic tumour volume (MTV) and microvessel density (MVD) and blood-borne metastasis in colorectal carcinoma.

METHODSThirty-six patients with CRC conformed by pathology underwent PET-CT examination before operation. SUVmax and MTV were obtained by PET VCRA software. The blood vessels were identified with CD34 immunohistochemical staining, and the MVD was recorded. The correlation between SUVmax and MTV with histological differentiation, T stage, MVD and blood-borne metastasis was analyzed.

RESULTSThe SUVmax, MTV and MVD in patients with blood-borne metastasis were 5.15 ± 5.41, (22.99 ± 18.63) cm³ and 14.17 ± 3.63, and were 10.65 ± 3.79, (16.95 ± 11.82) cm³ and 11.27 ± 3.69, respectively, in patients with non-blood-borne metastasis. The differences of SUVmax, MTV and MVD between blood-borne metastasis and non-blood-borne metastasis patients were statistically significant (all P > 0.05). Pearson correlation analysis found that there was no linear correlation between SUVmax and MVD, and the SUVmax was not statistically significant between high and low MVD groups (t = 0.919, P = 0.364). But there was a linear correlation between MTV and MVD (r = 0.621, P = 0.000), and the MTV was statistically significant between high and low MVD groups (t = 3.567, P = 0.001). The receiver-operating characteristic curves showed that MTV could be used to predict blood-borne metastasis of CRC, and the best cutoff value for MTV was 14.975 cm³, and the sensitivity, specificity, negative predictive value and positive predictive value were 85.7%, 54.5%, 72.3% and 64.2%, respectively. There were no significant relationships between SUVmax, MTV, MVD, blood-borne metastasis and histological differentiation (P > 0.05). With the increased T stage, the MTV, MVD and the probability of blood-borne metastasis were also increased (all P < 0.05).

CONCLUSIONSThere are correlations between MTV and MVD and blood-borne metastasis in CRC. The risk of blood-borne metastasis in patients with MTV > 14.975 cm³ is higher, and needs to take more effective intervention.

Colorectal Neoplasms ; blood supply ; diagnostic imaging ; pathology ; Fluorodeoxyglucose F18 ; Humans ; Microvessels ; pathology ; Multimodal Imaging ; Neoplasm Staging ; Neoplastic Cells, Circulating ; Positron-Emission Tomography ; ROC Curve ; Radiopharmaceuticals ; Sensitivity and Specificity ; Tomography, X-Ray Computed

This study examined the current changes of human ether-a-go-go-related gene (hERG) mutation derived from a LQT2 Chinese family with a highly penetrating phenotype.Mutation was identified and site-directed mutagenesis was performed to induce the mutation in wild-type (WT) hERG.WT　hERG and mutated V535M were cloned and transiently expressed in HEK293 cells.At the 48th and 72nd h after transfection,membrane currents were recorded using whole cell patch-clamp procedures.An A＞G transition at 1605 resulting in replacement of V535M was identified.Compared to WT,V535M mutation significantly decreased tail currents of hERG.At test potential of-40 mV after depolarizing at +50 mV,tail current densities were 83.35±7.06 pA/pF in WT and 50.38±7.74 pA/pF in V535M respectively (n=20,P＜0.01).Gating kinetics of hERG revealed that V1/2 of steady-state inactivation shifted to negative potential in the mutant (V1/2,V535M:-61.81±1.7 mV vs.V1/2,WT:-43.1±0.71mV).The time constant of recovery from inactivation was markedly prolonged in the mutant compared to WT among test potentials.V535M hERG mutation demonstrated markedly decreased tail current densities,which suggests that V535M is a new loss-of-function mutation of bERG channel responsible for LQT2.