Objective:To investigate the effect of chronic ethanol consumption on sensory information transmission in the cerebellar molecular layer and reveal the mechanism of chronic alcoholism on sensory information transmission and integration in the cerebellar cortex.Methods:Fifty healthy male ICR mice aged 6-8 weeks were randomly divided into saline group(control group)and ethanol consumption group(alcohol group) according to the random number table, with 25 mice in each group.The mice in alcohol group were injected intraperitoneally with 20% ethanol daily, while the mice in control group were injected with the same dose of normal saline. All mice were injected intraperitoneally once a day for 28 days.Through electrophysiological technology, patch-clamp amplifier and data acquisition software were used to record the changes in cerebellar molecular layer field potential of mice in the alcohol group and control group induced by sensory stimulation.Clampfit 10.3 software was used to record and analyze the electrophysiological data. SPSS 22.0 software was used for statistical analysis. Paired t-test and one-way ANOVA were used to analyze the differences before and after treatment. Results:After giving the stimulation of wind blowing, the amplitude of P1 in alcohol group was significantly higher than that in control group ((121.31±3.5)%, (97.2±2.7)%; t=26.08, P<0.05), and the area under the P1 curve (AUC) of the alcohol group was significantly lower than that of the control group ((127.1±4.2)%, (102.2±3.5)%; t=22.95, P<0.05). There was no significant difference in N1 amplitude between the two groups (P>0.05). When L-NNA, an inhibitor of nitric oxide synthase, was perfused into the brain surface of mice, the amplitude of P1 in alcohol group was significantly lower than that before administration ((76.2±4.8)%, (103.5±3.6)%; t=22.60, P<0.05), but there was no difference of the amplitude of P1 before administration and after elution ((101.5±4.6)%) ( t=1.70, P>0.05). After the L-NNA was perfused, the AUC of P1 was significantly lower than that before administration((72.4±5.6)%, (102.7±2.66)% ( t=24. 58, P<0.05), and there was no significant difference between before administration and after elution( (100.6±3.5)%, t=1.81, P>0.05). When L-NNA was perfused into the brain surface of mice, the amplitude of P1 in control group was (104.3±1.6)% and it had no differences compared with before administration(102.2±5.6)%, t=1.84, P>0.05) and after elution(102.5±4.5)%, t=1.92, P>0.05). And the AUC of P1 in control group after perfused L-NNA had no differences compared with before administration(103.5±2.6)%, (102.5±4.6)%) and after elution((101.9±3.7)%, t=0.99, 1.81, both P>0.05). When the mouse brain surface was perfused with NO donor SNAP, the amplitude of P1 in the control group was significantly higher than that before administration( (128.2±3.4)%, (103.5±2.6)%; t=28.89, P<0. 05) and there was no difference between before administration and after elution( (105.4±4.2)% , t=1.93, P>0.05). The AUC of P1((125.4±4.4)%) was higher than before administration((104.3±4.6)% , t=16.60, P<0.05) and there was no difference between before administration and after elution(103.5±4.2)%, t=0.65, P>0.05). Conclusion:Chronic ethanol consumption significantly enhances the inhibitory response, and the enhancement of inhibitory components stems from the activation of the NO signaling pathway.

Raw264.7 cells were incubated with receptor activator of NF-kappa B ligand (RANKL) and α-melanocyte stimulating hormone(α-MSH) for6 d.The amount of osteoclast cells were counted by tartrate resistant acid phosphatase staining and the acid phosphatase activity was assayed.The expressions of 5 melanocortin receptors (MCR) in Raw264.7 cells were determined by RT-PCR.The results showed that the number of osteoclasts in RANKL +α-MSH group was significantly increased compared with RANKL group (P ＜ 0.05),but there was no osteoclast formation in α-MSH group.Compared with control group and α-MSH group,the acid phosphatase activities were significantly increased in RANKL group and α-MSH+RANKL group (P＜0.05).All five MCRs were expressed in the Raw264.7 cells shown by RT-PCR.These results suggest that α-MSH may promote osteoclasts formation through RANK signaling pathway.

Objective:To investigate the relationship between serum retinol binding protein (RBP), stromal cell derived factor-1 (SDF-1) and renal function in patients with diabetic nephropathy (DKD).Methods:The patients with type 2 diabetes mellitus (T2DM) admitted to Yantai Affiliated Hospital of Binzhou Medical College from October 2017 to October 2020 were prospectively selected, 438 patients were divided into simple T2DM group ( n=276)and DKD group( n=162) according to the presence or absence of DKD, according to the ratio of urinary albinin/creatinine (UACR) were divided into normal( n=25), microalbuminuria ( n=75) and macroalbuminuria group ( n=62), according to the estimated glomerular filtration rate (eGFR) were divided into G1 stage ( n=28), G2 stage ( n=27), G3A + G3B stage ( n=35), G4 stage ( n=39)and G5 stages( n=33). The relationship between RBP, SDF-1 and renal function index UACR, serum uric acid (UA), blood urea nitrogen (BUN), β 2-microglobulin (β 2-MG) and serum creatinine (Scr) was analyzed. Measurement data of normal distribution were expressed as Mean± standard deviation ( Mean± SD). Independent sample t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups.Chi-square test was used to compare the enumeration data between groups. Receiver operating characteristic curve (ROC) was used to analyze the discriminant value of RBP and SDF-1 for DKD. Pearson was used for correlation analysis among indicators. Multivariate linear regression analysis was used to analyze the influencing factors of RBP. Results:In the DKD group, the duration of diabetes was longer, the levels of RBP, UACR, UA, BUN, β 2-MG, Scr were high, SDF-1 and eGFR were lower, with statistically significant differences compared with the simple T2DM group( P<0.05).The areas under the curve of RBP and SDF-1 to distinguish DKD were 0.903 and 0.868, and the optimal cut-off values was 70.71 mg/L and 5.69 ng/mL. With the increase of urinary albumin and clinical stage, the levels of RBP, UACR, UA, BUN, β 2-MG, Scr increased gradually, while SDF-1 and eGFR decreased gradually, and the differences were statistically significant ( P<0.05).RBP was positively correlated with UACR, UA, BUN, β 2-MG and Scr in DKD patients ( r=0.764, 0.787, 0.693, 0.577, 0.801, P<0.000 1), and negatively correlated with EGFR ( r=-0.782, P<0.000 1). SDF-1 was negatively correlated with UACR, UA, BUN, β 2-MG and Scr ( r=-0.744, -0.794, -0.666, -0.605, -0.820, P<0.000 1), and positively correlated with EGFR ( r=0.767, P<0.000 1). The multiple linear regression equation was RBP=29.852+ 0.007UACR+ 0.101UA+ 0.497BUN+ 0.034Scr-0.083eGFR ( P<0.001). Conclusion:RBP and SDF-1 have certain discriminant value for DKD patients in T2DM population, and the degree of DKD renal function injury is positively correlated with RBP and negatively correlated with SDF-1, the increase of UACR, UA, BUN, Scr and the decrease of eGFR are risk factors for the increase of RBP.