Objective : To explore the effect of liver-specific knockout of transcription factor EB (Tfeb) on high-fat diet ( HFD ) -induced hepatic steatosis in mice． Methods : Wild-type C57BL /6J mice and liver-specific Tfeb knockout C57BL /6J mice were fed with HFD or normal chow for 12 weeks，respectively，and then the serum tri- glyceride (TG) ，total cholesterol ( TC) ，monocyte chemoattractant protein-1 ( MCP-1 ) ，tumor necrosis factor-α (TNF-α) ，aspartate aminotransferase ( AST) and alanine transaminase ( ALT) were measured in each group of mice ; Western blot was used to detect Tfeb protein expression levels in liver tissues of mice in each group，HE stai- ning was used to monitor histopathological changes in liver tissues of mice in each group，oil red O staining was used to monitor lipid deposition in liver tissues of mice in each group，and F4 /80 fluorescence staining was used to monitor macrophage infiltration in liver tissues of mice in each group. Results : There was no expression of Tfeb gene in liver of liver-specific knockout Tfeb mice，suggesting that the effect of Tfeb gene knockout in liver tissue was better．Compared with the normal control group，the expression of Tfeb protein in the liver tissue of the model control group was down-regulated．Compared with the normal control group，both the HE staining results and the oil red O staining results showed that the liver specific Tfeb caused lipid deposition and liver lobule disorder in mice， which was similar to the liver changes in the model control mice，however，liver-specific knockout of Tfeb mice at 12 weeks of HFD had more severe liver lipid deposition and hepatic lobular structural disorder．The results of F4 / 80 fluorescence staining indicated that the specific knockout of liver Tfeb could aggravate the infiltration of macro- phages in the liver of mice induced by high-fat feeding．At the same time，the serological test results indicated that compared with the normal control group，the serum levels of TC，TG，TNF-α , MCP-1，AST and ALT in the liver- specific knockout Tfeb group and the model control group increased ，and these changes were further elevated in Tfeb knockout mice after HFD feeding． Conclusion Liver-specific knockout of Tfeb aggravates HFD-induced he- patic steatosis in mice.

Objective:To investigate the effects of body mass index (BMI) levels at different baseline on the risk of new-onset acute pancreatitis (AP).Methods:The subjects were from the Kailuan Study Cohort and divided into 3 groups according to baseline BMI levels: BMI<24 kg/m 2, normal weight; BMI 24-28 kg/m 2, overweight; BMI≥28 kg/m 2, obesity. The incidence of new-onset AP in these three groups was analyzed. The survival curve was plotted by Kaplan-Meier method, the cumulative incidence was calculated and tested by log-rank method. Multivariate Cox proportional hazards regression model was used to calculate HR of baseline BMI levels for AP. Results:A total of 123 841 subjects were included and followed up for (11.94±2.13) years, during which, 395 cases were found with AP. The incidence of AP was 2.67 per 10 000 person years in total population, and the incidences of AP were 2.20, 2.72 and 3.58 per 10 000 person-years in the normal, overweight and obesity groups, respectively. The cumulative incidences of AP was 0.32%, 0.40% and 0.49% in normal, overweight and obesity groups, respectively, which showed a significant inter-group difference by log-rank test ( χ2=13.17, P<0.01). The results of multivariable adjusted Cox proportional hazards regression model analysis indicated that obesity group ( HR=1.45, 95% CI: 1.10-1.92) had a higher risk for AP compared with the normal BMI group. The subgroup analyses by age and sex showed that compared with the normal weight group,the HRs for AP in the obesity group was 1.58(95% CI:1.14-2.19) and 1.40(95% CI:1.03-1.90) among subjects younger than 60 years old and male subjects, respectively. After excluded onset AP within two years from baseline,with a control group from normal weight,the results of multivariate Cox proportional hazards regression model analysis indicated that the AP in the obesity group was 1.60 (95% CI: 1.18-2.15). Conclusion:Obesity may increase the risk of developing AP, particularly among young and middle-aged men.

Objective To investigate the effects of fasting serum triglycerides (TG) levels at different baseline on the risk of new-onset acute pancreatitis (AP) in in-service and retired employees of Kailuan Group.Methods A total of 125 178 in-service and retired employees of Kailuan Group who received health check-ups from 2006 to 2009 and had no AP history but had complete TG data were prospectively enrolled.According to quantile level,the baseline serum fasting TG level of study subjects were divided into ＜1.01 mmol/L group (n=42 128),1.01 to 1.64 mmol/L group (n=41 711) and ＞ 1.64 mmol/L group (n=41 339).The incidence of new-onset AP of these three groups was analyzed.The survival curve was plotted by Kaplan-Meier method.The cumulative incidence rate was calculated and tested by log-rank method.And multivariate Cox proportional hazards regression model was performed to calculate hazard ratios (HR) of baseline fasting serum TG level for AP.Results After followed up for (7.36±1.23) years,a total of 193 cases of AP occurred.The incidences of AP in ＜1.01 mmol/L group,1.01 to 1.64 mmol/L group and ＞ 1.64 mmol/L group were 1.43 events/10 000 person-years,2.37 events/10 000 person-years and 2.49 events/10 000 person-years,respectively.The cumulative incidence rates of AP in ＜1.01 mmol/L group,1.01 to 1.64 mmol/L group and ＞1.64 mmol/L group were 0.10％ (44/42 128),0.18％ (73/41 711) and 0.18％ (76/41 339),respectively,and the difference was statistically significant (x2 =9.998,P=0.007).The results of multivariate Cox proportional hazards regression model analysis indicated that the risk of AP increased in 1.01 to 1.64 mmol/L group and ＞ 1.64 mmol/L group compared with that of ＜1.01 mmol/L group,HR and 95％ confidence interval (CI) were 1.56 (1.07 to 2.29) and 1.57 (1.06 to 2.32),respectively.After excluded onset AP within one year,with a control group of ＜1.01 mmol/L group,the results of multivariate Cox proportional hazards regression model analysis indicated that the HR and 95％CI for AP of 1.01 to 1.64 mmol/L group and ＞ 1.64 mmol/L group were 1.70 (1.11 to 2.58) and 1.69 (1.10 to 2.60),respectively.Conclusion Baseline fasting serum TG levels over 1.01 mmol/L may increase the risk of AP.

Objective:To investigate the influence of α cells and glucagon-like peptide l (GLP-1) in the function of β cells (INS-1 cells) in the rats,and to elucidate the possible mechanism of α cells and INS-1 cells transplantation in influencing hypoglycemia.Methods:The proliferation abilities of INS-1 cells after treated with 10％,20％ and 30％ islet α-cell conditioned medium and 0.03,0.30,3.00,30.0 mg · L-1 of GLP-1 were analyzed by MTT assay.The levels of insulin secretion of INS-1 cells after treated with 10％,20％,30％ α cells,α-cell conditioned medium and different concentrations of GLP-1 were analyzed by enzyme linked immunosorbent assay (ELISA).The concentrations of Ca2+ in INS-1 cells after treated with high glucose and GLP-1 were analyzed by laser confocal microscope.The expression levels of insulin protein after treated with different concentrations of islet α-cell conditioned medium and different concentrations of GLP-1 were detected by Western blotting methed.After the INS-1 cells,the mixture of INS-1 cells and α cells were transplanted into the left renal capsule of the nude mice,the blood glucose levels and the kidney morphology were observed.The levels of insulin/glucagon in the transplanted cells were detected by immunohistochemistry.Results:Compared with control group,both of α-cell conditioned media and GLP-1 promoted the INS-1 cell proliferation and insulin secretion (P ＜ 0.05).The laser confocal microscope results revealed that GLP-1 stimulated the increased intracellular Ca2+ concentration in INS-1 cells (P＜ 0.05).Compared with control group,there was no significant difference in the expression levels of insulin protein in the insulin-1 cells after treated with islet α cell conditioned medium and GLP-1 (P＞0.05).Compared with pre transplantation,the blood glucose level in the transplanted INS-1 cells was significantly decreased at 35 d after renal capsul trasplantation (P＜0.05),and even hypoglycemia presented renal capsular in the diabetic nude mice;the transplantation site was obviously swollen.However,the levels of blood glucose had no change of the diabetic rats after transplated with the mixture of INS-1 and α cells (P＜0.05).The expression of insulin and glucagon in the INS-1 transplanted cells were found by immunohistochemistry staining.Conclusion:Pancreatic islet α cells and their secretions can promote the INS-1 cell proliferation and insulin secretion,and the mixture of INS-1 cells and α cells transplanted under the renal capsule of the diabetic nude mice can reduce the hypoglycemic effect of INS-1 cell transplantation which might be related to the INS-1 cells that can express both of insulin and glucagon genes.

Objective:To observe the inhibitory effect of ischemic postconditioning (I-postC) on the apoptosis of renal cells after limb ischemia reperfusion(LIR) in the rats, and to investigate the possible mechanisms. Methods:Thirty SD rats were randomly divided control group, ischemia-reperfusion group(IR group) and I-postC group(n=10).4 h ischemia and 4 h reperfusion with the rubber band in two hind limbs of the rats were performed to establish the models.In control group, the rubber band around the limb was loose and the blood flow was not blocked.As for I-postC group, before perfusion, 5 min ischemia and 5 min reperfusion were performed in the rats and repeated 3 times named I-post C.The levels of blood creatinine (Cr), blood urea nitrogen(BUN) and C-reactive protein (CRP) in plasma of the rats in various groups were measured by automatic biochemistry analyzer.The expressions of Bcl-2 protein and Bax protein in renal were detected by immunohistochemical method,and its quantitative results were observed with automatic image analysis system and the ratio of Bcl-2/Bax was calculated.The apoptotic cells in kidney tissue were determined by terminal-deoxynucleotidy1 transferase-mediated d-UTP nick end labeling (TUNEL) under laser scanning confocal microscope(LSCM).The ultrastructures of kidney tissue were observed under electron microscope.Results:Compared with control group, the levels of Cr,BUN and CRP in plasma of the rats in IR group and I-postC group were increased(P<0.05 or P<0.01);compared with IR group, the levels of Cr, BUN and CRP in plasma of the rats in I-postC group were decreased(P<0.01).Compared with control group,the expression levels Bax and Bcl-2 in kidney tissue of the rats in IR group and I-postC group were significantly increased (P<0.05 or P<0.01),and the ratios of Bcl-2/Bax were reduced(P<0.05);compared with IR group,the expression level of Bax in kidney tissue of the rats in I-postC group was decreased (P<0.05),the expression level of Bcl-2 was increased(P<0.01),and the ratio of Bcl-2/Bax was increased(P<0.05).Under laser confocal microscope,the number of apoptotic cells in kidney tissue of the rats in IR group was increased compared with control group;the number of apoptotic cells in I-postC group was decreased compared with IR group.Under transmission electron microscope,the changes in IR group were found as follows: renal proximal convoluted tubule epithelial cell nucleus vacuoles,increased lysosome and dense particle deposition, some mitochondria crest fracture or fuzzy;irregular and fusion, glomerular podocyte protuberance,mitochondrial cristae fracture and reducetion with vacuoles, rough endoplasmic reticulum expansion;the damage levels of renal tubular epithelial cells and glomerulus in I-postC group were improved compared with IR group.Conclusion: Limb ischemia reperfusion can induce the apoptosis of renal cells, I-postC can inhibit the apoptosis of renal cells,and it would be helpful to improve the kidney function.

Objective:To observe the effects of ischemic postconditioning (I-postC)on the lung injury after limb ischemia reperfusion (LIR)in the rats,and to investigate the protective effect and the possible mechanisms. Methods:24 Wistar rats were randomly divided into control group, ischemia-reperfusion group (IR group)and I-postC group (n=8 ). Referring to routine method in our department, the model rats which underwent 4 h ischemia and 4 h reperfusion of hind limbs were made.In control group,the rubber band around the limb was loose and the blood flow was not blocked. In I-postC group, before reperfusion, ischemia 5 min and reperfusion 5 min were performed in the rats,repeated for 3 times and then perfusion 4 h was taken,The blood and lung tissue from every rat were taken accurately. The percentages of CD1 8 positive cells in peripheral blood,the levels of soluble intercellular adhesion molecule-1 (sICAM-1)and P-selectin in plasma,the myeloperoxidase (MPO)activities in lung tissue,the levels of intercellular adhesion molecule-1 (ICAM-1)and P-selectin in lung tissue of the rats in various groups were detected. The partial pressure of oxygen (PaO2 ) and partial pressure of carbon dioxide (PaCO2 )were measured.The morphological changes of lung tissue under light and electron microscopes were observed.Results:Compared with control group,the percentage of CD18 positive cells and the levels of sICAM-1 and P-selectin of the rats in IR groups were increased (P<0.01);PaO2 and PaCO2 were decreased significantly;the MPO activity in lung tissue was also significantly increased (P<0.01).The HE staining results showed lung interstitial vascular dilation, congestion, PMN infiltration, the increased gap blood vessel, alveolar septal thickening,alveolar exudation, bronchial epithelial cell shedding and necrosis of the rats in IR group. Compared with IR group,the values of biochemical indicators mentioned above were decreased obviously (P<0.01);PaO2 and PaCO2 were increased significantly (P<0.01);the activities of inflammatory factors in plasma and lung tissue were decreased (P < 0.01 ); the pathological changes of lung damage were improved significantly. Conclusion:I-postC can reduce the lung injury after LIR in the rats,and the mechanism may be related to inhibiting the inflammatory reaction.

Objective To observe the effects of ischemic postconditioning (I-postC) on lung injury after limb ischemia reperfusion (LIR) in rats, and to investigate the protective effect and the mechanisms. Methods Twenty-four Wistar rats were divided into three groups:control group (group Control), ischemia-reperfusion group (group IR) and ischemic postcondi?tioning group (group I-postC). Referring to routine method in our department, the model rats underwent 4-hour ischemia and 4-hour reperfusion of hind limbs were made. In group Control, the rubber band around the limb was loose,which did not block the blood flow. Rats in group I-postC were given repeated 3 times of 5 min ischemia-5 min reperfusion, and then did perfusion 4 h before reperfusion. The blood and lung samples were collected for detecting arterial gas of partial pressure of oxygen [p(O2)] and partial pressure of carbon dioxide [p(CO2)]. The plasma and lung tissue levels of malondialdehyde (MDA), superoxide dismutase (SOD) and xanthine oxidase (XOD) were detected. The morphological changes of lung tissue were ob?served under light microscope and electron microscope. Results It was found that after suffering from ischemia-reperfu?sion, levels of p(O2) and p(CO2) decreased significantly. The activity of SOD in plasma and lung tissues decreased, but XOD and MDA increased significantly (P<0.05). With microscope, lung interstitial vascular dilation, infiltration of neutrophils, the width of the alveolar space, alveolar septal thickening and alveolar exudate were found. Compared with IR group, it was found that p(O2) and p(CO2) increased significantly in group I-postC. The activity of SOD in plasma and lung tissues in?creased, but XOD and MDA decreased significantly(P<0.05). The mild damage of pathological changes were found. Conclu?sion Ischemic postconditioning can reduce the lung injury after limb ischemia reperfusion in rats, which may be related to the inhibition of lipid peroxidation.

Objective To investigate the imaging features of hippocampus and entorhinal cortex in the normal,mild cognitive impairment(MCI) and Alzheimer's disease (AD),and explore the value of diagnosing MCI and AD by using the method of MRI measuring the volume of hippocampus and entorhinal cortex.Method One hundred and twenty-two people including 42 cases of MCI,38 cases of AD,and 42 cases of noroal cognition(NC) were selected as our subjects from health examination persons both in hospital and outpatient service.All were performed general examination and neuropsychological scale evaluation.The volume of hippocampus and entorhinal cortex were measured by using MRI.The correlation between the volumetric changes of hippocampus and entorhinal cortex with scores of mini-mental state examination (MMSE) and Montreal Cognitive Assessment(MoCA) was analyzed.Results The volume of hippocampus and entorhinal cortex in the MCI group,AD group and NC group were (6.29 ± 1.13)cm3 and (2.71 ± 0.51) cm3,(6.27 ± 1.11) cm3 and (2.09 ±0.68) cm3,(7.01 ±0.92) cm3 and (3.12 ±0.34) cm3 respectively.The volume of MCI group was obviously smaller than that of NC group (P ＜ 0.05).The volume of AD group was smaller than that of NC group(P ＜0.01).The volume of AD group was obviously smaller than that of MCI group(P ＜0.01).There was positive correlation between hippocampus volume,the volume of entorhinal cortex and MMSE scores (r =0.770,0.811 ; P ＜ 0.01).Meanwhile,hippocampal volume,volume of entorhinal cortex were positive correlated with MoCA (r =0.810,0.842; P ＜ 0.01).Conclusion The atrophy of entorhinal cortex and hippocampus is closely related to cognitive disorder.The MRI measuring of the volume of entorhinal cortex and hippocampus has a potential value in diagnosing and distinguishing of MCI and NC.

Objective To investigate the influence and mechanism of erythropoietin (EPO) in renal blood flow after limb ischemia reperfusion (LIR). Methods Thirty male SD rats were randomly divided into control group, LIR group and EPO+LIR group with ten in each group. The values of renal blood flow, plasma creatinine (Cr), urea nitrogen (BUN) content in plasma, kidney tissue wet to dry ratio (W/D), nitric oxide (NO), nitric oxide synthase (NOS) and endothelin-1 (ET-1) in re-nal tissue were detected in three groups. The immunohistochemistry assay was used to detect the expression of intercellular adhesion molecule (ICAM-1) and vascular cell adhesion molecule (VCAM-1) in renal tissue. The morphological changes of renal tissue were observed with light microscope. Results The renal blood flow was significantly decreased, while the val-ues of Cr, BUN, W/D, NO, ET-1, NOS, expressions of ICAM-1 and VCAM-1 was significantly increased in LIR group than those of control group (P<0.05). Broaden interstitial and infiltration of inflammatory cells were observed in the renal tissue under light microscope. In the EPO+LIR group, the renal blood flow increased, the values of Cr, BUN, W/D, NO, ET-1 and NOS, expressions of ICAM-1 and VCAM-1 decreased significantly compared with those of LIR group (P<0.05). The patho-logical changes were alleviated in EPO+LIR group. Conclusion EPO can improve renal function, increase renal blood flow in rats after LIR. The mechanism may be related to the decreased edema, changed renal vasomotor function and decreased in-flammation.

We developed a three-dimensional mini-type permanent magnetic resonance imaging (MRI) device in our lab. The purposes of this study were (1) for further development of MRI technologies, (2) for support of broadening practices of animal test modeling in medical research, and (3) for training more specialists from colleges or universities in the field of MRI. This paper describes the research and development at our lab(s), especially stressing on the design of the main magnet, the gradient coil and the radio frequency coil. In addition, the specific methodologies used in our lab(s) and the related data are emphasized. The 3D MRI technologies have met the needs of using small animals, super thin sections of live animal body and high imaging resolutions. MRI images of mice head and abdominal have been obtained successfully by using the imager that we developed. The imaging results and analyses have also been discussed.
Animals ; Equipment Design ; Imaging, Three-Dimensional ; instrumentation ; Magnetic Resonance Imaging ; instrumentation ; methods ; Mice