Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.

Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.

Through serial problem-oriented teaching method(S-PBL),problem based learning-case-based-case based learn-ing(PBL-CBL)comprehensive teaching method,industry-university-research case integration teaching,cutting-edge literature review and other teaching methods,students'attention to learning immunology knowledge is grasped and students'learning efficiency is im-proved;combined with immunology experiment,enhance students'understanding of classroom teaching content;through professional practice,strengthen students'understanding of the development process,production process and market demand of immune products,and enhance students'interest and initiative in learning and consolidating immunology knowledge;through the introduction of major research achievements such as the Nobel Prize in immunology,students'interest and motivation in exploring innovative content in the field of immunology research will be cultivated.Through the above teaching reform,we will cultivate a new generation of biological people who have a solid knowledge of immunology,have the potential to develop immune products,have the sense of innovation and entrepreneurship,and have responsibility and responsibility.

Objective:To use metagenomic sequencing to compare the differences in intestinal microbiota species and metabolic pathways in patients with hepatitis B cirrhosis with or without ascites and further explore the correlation between the differential microbiota and clinical indicators and metabolic pathways.Methods:20 hepatitis B cirrhosis cases [10 without ascites (HBLC-WOA), 10 with ascites (HBLC-WA), and 5 healthy controls (HC)] were selected from the previously studied 16S rRNA samples. Metagenome sequencing was performed on the intestinal microbiota samples. The Kruskal-Wallis rank sum test and Spearman test were used to identify and analyse differential intestinal microbiota populations, metabolic pathways, and their correlations.Results:(1) The overall structure of the intestinal microbiota differed significantly among the three groups ( R = 0.19, P = 0.018). The HC group had the largest abundance of Firmicutes and the lowest abundance of Proteobacteria at the genus level. Firmicutes abundance was significantly decreased ( Pfdr < 0.01), while Proteobacteria abundance was significantly increased ( Pfdr < 0.01) in patients with cirrhosis accompanied by ascites; (2) LEfSe analysis revealed that 29 intestinal microbiota (18 in the HBLC-WA group and 11 in the HBLC-WOA group) played a significant role in the disease group. The unclassified Enterobacteriaceae and Klebsiella species in the HBLC-WA group and Enterobacteriaceae in the HBLC-WOA group were positively correlated with the Child-Turcotte-Pugh (CTP) score, prothrombin time, and international normalized ratio score and negatively correlated with albumin and hemoglobin levels ( P < 0.05). Escherichia and Shigella in the HBLC-WA group were positively correlated with CTP scores ( P < 0.05); (3) The correlation analysis results between the KEGG pathway and 29 specific intestinal microbiota revealed that Enterobacteriaceae and arachidonic acid, α-linolenic acid, glycerolipid metabolism, and fatty acid degradation were positively correlated in the lipid metabolism pathway, while most Enterobacteriaceae were positively correlated with branched-chain amino acid degradation and negatively correlated with aromatic amino acid biosynthesis in the amino acid metabolic pathway. Conclusion:A significant increment of Enterobacteriaceae in the intestines of HBLC-WA patients influenced hepatic reserve function and was associated with amino acid and lipid metabolic pathways. Therefore, attention should be paid to controlling the intestinal microbiota to prevent complications and improve the prognosis in patients with hepatitis B cirrhosis, especially in those with ascites.

Background: Disrupted circadian rhythms have been associated with the development of irritable bowel syndrome (IBS). In some IBS patients, the symptoms may present with circadian fluctuations. Enterochromaffin cells (EC cells) and tryptophan hydroxylase 1 (TPH1) - 5 - hydroxytryptamine (5 - HT) signaling pathway are currently recognized as the key pathophysiological mechanism of IBS. Aims: To explore whether Bmal1, the core circadian clock gene, is involved in the occurrence of IBS by regulating TPH1-5-HT signaling pathway in EC cells. Methods: Normal Sprague-Dawley (SD) rats and IBS-model SD rats, as well as wild type (WT) and intestine-specific Bmal1 knockout (Bmal1

Objective:To investigate the changes of dynamic functional connectivity between the default mode network (DMN) and executive control network (ECN) in the resting state in patients with alcohol use disorder (AUD).Methods:From September 2018 to June 2019, 23 cases of AUD group and 24 cases of healthy control (HC) group matched with age, gender, education level and handedness were collected at Renmin Hospital of Wuhan University. Mini-mental state examination (MMSE) and Michigan alcoholism screening test (MAST) were performed in all subjects for cognition and alcohol dependence score. All the subjects underwent T 1WI-3D structural imaging and resting state functional MRI (rs-fMRI) examination. Group spatial independent component analysis (ICA) was used to select the independent components of DMN and ECN. Then dynamic changes in the functional connectivity between the DMN and the DMN were obtained by sliding window approach and clustering method. Finally, the independent sample t test was used to compare the difference of general clinical data between the two groups, the linear correlation analysis was conducted in the parameter value and MMSE and MAST scores. Results:Compared with the HC group, the static functional connectivity analysis showed that the precuneus and posterior cingulate gyrus of the DMN were enhanced in the AUD group ( P=0.016, t=2.496). The DMN and ECN functional connectivity showed four different brain activity states, the proportion of state1 increased by 6.81% and state2 decreased by 6.83% in the AUD group, state3 and state4 were relatively stable. In state1, the internal functional connectivity of the DMN in the AUD group was enhanced, while the functional connectivity between DMN and ECN was mainly enhanced. In state2, the internal functional connectivity of the ECN was enhanced, and the connectivity between the DMN and ECN was mainly weakened. The mean dwell of state2 in the AUD group was negatively correlated with the MAST score ( r=-0.433, P=0.039). Conclusions:Dynamic functional connectivity patterns between DMN and ECN have been changed in patients with AUD. Dynamic functional connectivity can reveal transient changes in brain activity, which can provide certain imaging evidence for finding changes in AUD deep brain activity.

Objective To study the effect of aralia total saponins on renal function of type 2 diabetic mice, and its effect on the Bax and Bcl-2 protein in renal tissues, in order to provide some references for the development of aralia total saponins. Methods The mice were divided into the normal group, model group, positive control group, low, medium and high dose aralia total saponins group by random number method. Except the normal group, the others were received with high-fat diet for one month+one-time large dose of streptozotocin (STZ) to induce type 2 diabetic model, and then the mice in the normal group and the model group were intragastrically administered with the same volume of normal saline, and the mice in the positive control group was given 1 mg/kg of benazepril solution, and the low, medium and high dose groups were given 30, 60, 120 mg/kg aralia total saponins. The body weight of 1 ml/kg mice was intragastrically administered once a day. After treatment for 6 weeks they were sacrificed, and the serum insulin, and SOD and MDA levels were measured, the urine creatinine (Cr), urea nitrogen (UN), and uric acid (UA) levels were also measured. The immunohistochemistry and Western blot were used to detect the Bax and Bcl-2 protein expression in kidney tissues. Results Compared with the model group, the blood glucose and insulin resistance index in the low, medium and high doses aralia total saponins group were significantly decreased (P<0.05); the levels of urine UN, Cr and UA significantly decreased (P<0.05); The serum SOD level increased and the MDA level significantly decreased (P<0.05). The average gray value of Bcl-2 increased (92.26 ± 11.36, 107.17 ± 9.26, 132.65 ± 8.45 vs. 56.42 ± 16.24) in kidney tissue. The average gray value of Bax (152.62 ± 9.86, 124.48 ± 10.36, 92.29 ± 10.10 vs. 171.38 ± 15.18) significantly decreased ( P<0.05); Bax protein (0.81 ± 0.06, 0.75 ± 0.07, 0.52 ± 0.09 vs. 2.02 ± 0.09) significantly decreased, but Bcl-2 protein (0.92 ± 0.08, 0.94 ± 0.12, 1.27 ± 0.07 vs. 0.30 ± 0.09) significantly increased (P<0.05). Conclusions The aralia total saponins can reduce blood sugar levels, meanwhile improve renal function in type 2 diabetic mice. The mechanism may be may be that aralia total saponins could improve the antioxidant capacity and inhibition of renal cell apoptosis.

Objective To observe the effect of aralia saponins on the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in the kidney of diabetic nephropathy mice.Methods After 10 days of adaptive feeding,90 clean Kunming mice were randomly divided into the normal group (n =10) and 5 model groups (model group,positive drug benazepril intervention group,aralia saponins low,middle and high doses treatment groups).Excepted the normal group,the kidney damage model of type 2 diabetes mellitus in mice was induced by high-fat and high-sugar diet for one month plus disposable streptozotocin (STZ).The model was successfully constructed and killed after 6 weeks of treatment.A total of 25 mice failed to establish the model.And totally 55 mice were randomly divided into 5 groups with 11 mice in each group.The serum changes of blood urea nitrogen (BUN),serum creatinine (SCr),insulin,inflammatory factors interleukin-1α (IL-1α),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in each group were detected.The expression of vascular endothelial growth factor and MMP-9 protein in renal tissue were detected by immunohistochemistry and Western blot.Results Compared with the normal group,the levels of blood glucose,insulin,BUN,SCr,IL-1α,IL-6 and TNF-α in the model group were significantly increased (P ＜ 0.05).The above indexes were decreased in positive drug group and aralia saponins treatment groups.The contents of insulin,BUN and SCr in the high dose of aralia saponins group were significantly lower than those parameters in benazepril group (P ＜ 0.05).In addition,the contents of blood glucose,IL-1α,IL-6 and TNF-α in the three dose aralia saponins groups were significantly lower than those parameters in the benazepril group (P ＜ 0.05).Compared with the normal group,the expression level of VEGF protein in the model group was significantly higher (P ＜ 0.05),and the expression level of MMP-9 protein was significantly lower (P ＜ 0.05).Compared with the model group,both benazepril and aralia saponins can reduce VEGF (P ＜ 0.05),increase MMP-9 (P ＜ 0.05).In addition for VEGF and MMP-9,the high dose of aralia saponins group and benazepril group was basically same.Conclusions Aralia saponins can significantly reduce blood glucose,insulin and serum inflammatory factors,while downregnlate VEGF and increase MMP-9 protein levels,thereby protecting the kidneys of diabetic nephropathy mice.

Objective To investigate the protective effect of aralia total saponins on renal injury induced by streptozotocin (STZ) in type 2 diabetic mice, meanwhile to explore its protective mechanism. Methods Fifty male Kunming mice were randomly divided into the normal group, the model group and the aralia total saponins low, middle and high does groups. All the rats were given high fat diet 8 weeks and then received STZ 45 mg/kg to built type 2 diabetic mice model, except the noraml group. After the models establishment,the aralia total saponins low, middle and high does groups were given the aralia total saponins 30, 60, 120 mg/kg treatment, andthe normal group and the model group were given the equal normal saline, once each day. After 4th and 8th week administration, the urinary protein levels of 24 h in each group were detected. After the last treatment, all the mice were sacrificed to detected the changes of blood glucose, insulin and inflammatory related factors. Immunohistochemistry, Western blotting and real-time quantitative PCR were used to observe the expression of TLR2 and TLR2 in the kidney tissue. Results Compared with the model group, the low, middle and high does groups in 24-hour proteinuria, blood glucose, insulin resistance index decreased (P<0.05), the insulin increased(P<0.05). The serum TNF-α (16.66 ± 0.20 ng/L, 14.49 ± 0.27 ng/L, 13.52 ± 0.22 ng/L vs.20.33 ± 0.56 ng/L),IL-1β(0.46 ± 0.04 ng/L,0.44 ± 0.04 ng/L,0.37 ± 0.04 ng/L vs.0.55 ± 0.05 ng/L),NF-κB (28.71 ± 6.14 ng/L, 26.26 ± 5.48 ng/L, 25.69 ± 5.61 ng/L vs. 36.55 ± 8.90 ng/L) significantly decreased (P<0.05).The kidney TLR2 mRNA(1.92 ± 0.18,1.46 ± 0.23,1.28 ± 0.21 vs.2.69 ± 0.22),TLR4 mRNA(2.20 ± 0.19,2.08 ± 0.27,1.57 ± 0.22 vs.2.78 ± 0.23),TLR2 porteins(0.82 ± 0.11,0.52 ± 0.06,0.44 ± 0.07 vs.0.77 ± 0.13),TLR4 proteins(0.52 ± 0.04,0.42 ± 0.09,0.26 ± 0.06 vs.0.86 ± 0.12)significantly decreased(P<0.05). Conclusions The aralia total saponins can significantly reduce the blood glucose, insulin resistance index and 24-hour urinary protein in type 2 diabetic nephropathy of mice, increase the insulin, and analyzing its mechanism may be total saponins can inhibit the expression of TLR2 and TLR4 protein in the kidney, and further reduce the inflammatory response.

Objective To investigate the effect of bitter gourd saponins on the salt -sensitive hypertension caused kidney damage in rats ,and analyze the mechanism of its therapeutic effect .Methods 50 SD rats were fed for 10 days with normal diet ,and then based on random number method ,the rats were randomly divided into 5 groups:the normal control group,model control group and the low total saponins bitter,medium and high dose treatment group,10 rats in each group.Then,the rats in the normal control group and the model control group were given 1mL· kg-1 · d-1 normal saline,and the rats in bitter gourd saponins groups were given high salt diet 8 weeks to establish the model .After 4 weeks,the total saponins of bitter gourd treatment groups were given 10,20 and 40mg· kg-1· d-1.8 weeks later, the rats were sacrificed and the renal pathology was detected by HE staining .The changes of blood pressure , heart rate,urinary function and blood renal function were also analyzed .At last,Western blot and semi -quantitative PCR were used to detect the endothelin -1 (ET-1) in renal tissue.Results For the systolic blood pressure,in whole of treatment,the normal control group maintained at about 120 mmHg, while the model control group maintained at 170mmHg,which of the bitter gourd saponins three doses groups compared to the model control group was significantly lower (t=1.765,1.982,2.126,all P<0.05),further improved.And the heart rate had no statistically significant difference among the groups(all P>0.05).Compared with the normal control group,the blood urea nitrogen (BUN), creatinine (Cr),uric acid (UA),urine volume,urinary protein and N -acetyl beta -D-Glucosaminidase (NAG) levels in the model control group were significantly increased (t=28.703,33.932,29.298,4.695,10.989,10.871, all P<0.05),which in the bitter gourd saponins treatment groups were significantly decreased (all P<0.05).HE staining showed that the rats in the normal group were normal ,and the model control group had obvious glomerular sclerosis and renal interstitial fibrosis , after bitter gourd saponins treatment , the condition was significantly improved .Immu-nohistochemistry,Western blot and semi-quantitative PCR showed that normal control group had almost a little expression of ET-1,and compared with the normal control group ,which of the model control group was significantly increased ( t=14.650,11.387,all P<0.05),and the ET-1 expression in the bitter gourd saponins treatment groups significantly decreased.Conclusion The bitter gourd saponins can significantly improve the symptoms of hypertension and renal damage induced by high salt diet in rats ,which may be related with regulation of ET -1 expression in renal tissue .